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Home , Bahria University, Baqai Medical University, University of Karachi, Ziauddin University

New Horizons in Translational 3 (2016) 175–219

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New Horizons in Translational Medicine

journal homepage: www.elsevier.com/locate/nhtm

Abstracts: Joint 2nd annual conference of Ziauddin , HEC with European Society for Translational Medicine (EUSTM) International Conference on Translational Medicine: From discovery to care, 1–3 February 2016, , Pakistan

Nikhat Ahmed Siddiqui a,n,1, Aamir Shahzad b,c a Dean Research and Department of , , Pakistan b Departments of Neurology and , University of Colorado School of Medicine, USA c European Society for Translational Medicine (EUSTM), Austria

Oral presentations relation with TC incidence. Relation of thyroid cancer and anthropometric factors in Expression alterations of metabolic proteins in schizo- Pakistan phrenic brain regions Sohail Awan1*, Adnan Ali1, Ramsha Shakeel2, Sadia Wasay2 and Ayesha Khan1, Beena Hasan1, Shamim Mushtaq2, Abdul R Asif3and Ahmed Rahim1 Nikhat Ahmed1,2 1The Hospital, Karachi, Pakistan, 2Dow 1Department of Biochemistry, , Karachi, Pa- University of Health , Karachi, Pakistan. kistan, 2Department of Clinical , Ziauddin University, Kar- Introduction and objective: We have investigated the re- achi, Pakistan,3University Medical Center Goettingen, Goettingen, lationship of thyroid cancer (TC) and anthropometric factors in Germany. Pakistani population as this association remains to be Introduction: Schizophrenia (SZ), a multifactorial disorder controversial. exhibits diverse neuropathological aberrations including defective Methods: A total of 172 TC patients admitted at Aga Khan neurotransmission system, neuroanatomical abnormalities and University Hospital (AKUH) from 2000 to 2014 were included. impaired synaptic connectivity. Along with these complications Patient’s demographics, tumor subtypes, body weight, height, there is surfacing evidence that metabolic dysfunction and energy body mass index (BMI), body surface area (BSA) and body fat deficit may also contribute to its pathophysiology. percentage (BF%) were calculated. Methods: Present study entails, differential proteomic analysis Results and discussion: Out of 221 patients, 172 patients were of autopsied schizophrenic brain substantia nigra, cortex, and included in this study. Mean age was 43 years715.40. Among all hippocampus (n¼7) by using two dimensional electrophoresis subtypes of TC, papillary remains the most common (72.1%), fol- (2DE) followed by Orbitrap mass spectrometry identification. lowed by medullary (10.5%), follicular (7.6%) and others (9.9%). Differential expression was validated by western blot, while In Excess BMI 423 kg/m2, BF%, BSA, weight and height were sig- Silico analysis was applied for functional interactive role in nificantly associated with TC incidence in overall population (p- metabolism. valueo0.01*). In women, BF% was found to be in obese range Results: Proteome mapping of SZ patients revealed more pro- (mean¼35.07%77.6, p-valuer0.01*). In men, height 4160 cm found alterations in SN protein expression than other brain re- and weight 460 kg were positively associated with TC. No sig- gions compared to controls. SN exhibits differential expression of nificant relation was seen between gender and BMI. six proteins, enriched in several energy metabolic pathways in- Conclusions: Excess BMI 423 kg/m2 and age 440years were cluding decreased expression of phosphoglyceratemutase 1, ATP positively associated with TC. In women, increased BF% and in synthase subunit d, mitochondrial and malate dehydrogenase cy- men, height 4160 cm and weight 460 kg had a significant toplasmic, while increased expression of glyceraldehyde-3-phos- phate dehydrogenase, 4-trime-thylaminobutyraldehyde dehy- drogenase, and alcohol dehydrogenase were found as compare to n Corresponding author. normal brain. Cortex and hippocampus also portray significant E-mail address: [email protected] (N.A. Siddiqui). fi 1 Please note that the selected abstracts were chosen by the editors and this variations in expression patterns of ve proteins across brain of publication does not include all abstracts presented at the congress. schizophrenia patients. http://dx.doi.org/10.1016/j.nhtm.2016.11.001 2307-5023/& 2016 European Society for Translational Medicine. Published by Elsevier Ltd. All rights reserved. 176 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

Conclusions: Identification of differentially expressed meta- cellular networks. An important observation was a complex com- bolic proteins provides an insight for a better understanding of prised of novel interacting partners including glyceral-dehyde-3- schizophrenia pathophysiology. Further characterization, will PO4 dehydrogenase, actin cytoplasmic, microtubule associated elucidate the functional mechanisms underlying defective meta- protein 1B and glial fibrillary acidic protein. Further the interaction bolic pathways, which will provide promise for future therapeutic between actin and GAPDH confirmed by co-immunoprecipitation developments in the treatment of schizophrenia. has not been established before in AD. Analyses of pre-mRNA splicing machinery by UV-induced Conclusion: In accordance with the previous findings of protein-RNA cross-linking and mass spectrometry GAPDH and actin differential expression and post translational Uzma Zaman1and Henning Urlaub1 modification, our results suggest interesting implications of these Bioanalytical Mass Spectrometry Group, Max Planck Institute for two proteins in AD. Biophysical Chemistry, Goettingen, Germany. Detection of high risk human papilloma virus (HPV) geno- Introduction and objectives: The human genes that encode types 16/18 in oral lesions of tobacco chewers in Pakistan proteins undergo pre-mRNA splicing, an essential step of gene Zile Rubab1*, Saeeda Baig1and Mohammad Haris Lucky1 expression in which noncoding sequences are removed and coding 1Department of Biochemistry, Ziauddin Medical , Ziauddin sequences are ligated together. Unraveling splicing mechanism at University, Karachi, Pakistan the molecular level is not only important for understanding gene Introduction and objective: The Human Papilloma Virus expression, but it is also of medical relevance, as aberrant pre- (HPV) has evolved as a new culprit of malignant and premalignant mRNA splicing is the basis of many human diseases or contributes oral lesions. The objective of this study was to find out the fre- to their severity. Although methods based on high-throughput quency of HPV and its high risk genotypes in different lesions of sequencing have advanced our ability to identify the specificRNAs oral cavity of tobacco chewers. bound by a particular protein, there is a need for precise and Methods: From 492 subjects (421 males and 71 females), 20 ml systematic ways to identify RNA interaction sites on proteins. of oral rinse sample was collected after obtaining an informed Methods: We have developed an experimental workflow consent. Normal subjects with no chewing habits (250) including combining UV-induced cross-linking and high-resolution mass 135 males and 115 females were also taken from same setting. spectrometry for the identification of protein-RNA cross-linking Gentle brushings over the lesions with the help of dental floss sites. Purified complexes were UV irradiated at 254 nm and pro- brush and stored at 4 °C until DNA extraction. DNA was extracted teins therein hydrolyzed with trypsin. Peptide-RNA cross-links and PCR was performed using HPV consensus primers Gp5þ/ were enriched by TiO2 chromatography and analyzed by ESI-MS. Gp6þ and HPV 16, 18 specific primers for genotyping. Categorical Data analysis for identification of putative cross-links was per- data was calculated as frequencies and percentages. formed by novel RNPxl pipeline. Results: Oral pre-malignant lesions were present in 421 (86%) Results: The workflow not only enables the systematic identi- males and 71 (14%) females having leukoplakia (173, 35%), ery- fication of cross-linked peptides but also amino-acid residues in throplakia (60, 12%), submucous fibrosis (192, 39%) and L/E (67, RNA-interacting domains of proteins. 14%). Total number of HPV positives were 128 (26%), having HPV Conclusions: Successful identification of several cross-linking 16 (13%) and HPV 18 (11%) whereas, 76% had other genotypes. sites from human and yeast splicing complexes make it a pro- Among submucous fibrosis 82 (46%) were HPV positive. Out of mising technique to study any RNA-protein complex of interest. total 128 HPV patients 92% were males and 8% were females. All Crosstalk between cytoskeletal and metabolic proteins: A controls were found to be HPV negative. roadmap to ad pathology Conclusion: Frequency of HPV was found high (26%) in oral Beena Hasan1,2, Ayesha Khan1, Christof Lenz3, Abdul R Asif3and lesions with HPV16/18 as 13% and 11% respectively. The patients Nikhat Ahmed4* with submucous fibrosis are at greater risk of having HPV. Other 1Neurochemistry Research Laboratory, Department of Biochem- HPV genotypes causing premalignant lesions require further istry, University of Karachi, Pakistan,2The Karachi Institute of Bio- investigation. and Genetic (KIBGE), University of Karachi, Diagnostic, prognostic and predictive value of microRNA-21 Pakistan,3Institute of Clinical Chemistry, University Medical Center in breast cancer patients, their daughters and healthy Goettingen, Germany,4Research Department, Ziauddin University, individuals Karachi, Pakistan. Ambreen Usmani1, Amir Ali Shoro1, Zahida Memon1and Mehvish Introduction and objectives: Cellular activities are based on Hussain1 complex networks of protein interactions. Identification and in- 1Bharia University and Ziauddin University, Karachi, Pakistan. teraction analysis of associated protein partners is the basis of Introduction: MicroRNA-21 (miR-21) located on 17q23.1 has many biological studies, determining the key etiological factors. shown high expression in breast cancer. This gene has anti- Alzheimer’s disease, a pathologically complex and multifactorial apoptotic ability and causes tumor cell growth. It is also normally disease is the most common cause of dementia. It is characterized involved in functions such as signal transduction pathways ef- by a progressive decline in memory and cognitive functions. fecting normal cell growth and differentiation. Aberration of multiple pathways is observed in Alzheimer’s dis- Objective: To determine the presence of miR-21 in the serum ease including metabolic pathways which subsequently alter the levels of breast cancer patients, their daughters and healthy allied cytoskeletal system. This study is designed to unveil the individuals. cytoskeleton and metabolic proteins interaction in AD and inter- Methods: A total of 132 subjects were recruited: 50 breast pret their association. cancer (cases), 50 age-matched healthy individuals (control A) and Methods: Human autopsied brain tissue samples from normal 32 daughters of index cases (control B). The sampling technique age matched control (n¼6) and AD patients (n¼5) were procured. used for cases was random sampling. Controls were selected by Two dimensional Blue Native/SDS PAGE combined with Orbitrap purposive method. Serum tests were run on qRT PCR and mass spectrometry analysis was accomplished to determine the threshold cycle was determined and fold change calculated. interacting cytoskeleton and metabolic proteins. The interaction Results: Fisher exact test was used when analyzing between was further confirmed by co-immunoprecipitation. cancer patients and healthy patients or with their daughters. Results: A total of thirteen protein complexes were obtained on Normality of continuous variables was assessed by Shapiro Wilk’s BN-PAGE which resolved into many crucial proteins of different test. While comparing the same between two groups, student t- N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 177 test and Mann-Whitney test was used. P-valuer0.05 was con- Introduction and objectives: Acute kidney injury (AKI) is a sidered significant. MiR-21 was significantly higher in cases as complex disorder manifested by rapid loss of renal function. AKI compared to control A and B (p-value¼0.001). However control B has strong influence on expression of renal specific genes which showed significant gene expression as compared to control A (p- may play central roles in key cellular pathways and targeting these value¼0.001). The 2^ct formula was applied to compare the fold pathways may turn out to be effective therapeutic option. Trans- chain between the groups. The cases were also divided into ER, PR plantation of mesenchymal stem cells (MSCs) has been shown to and HER2 positive and negative cases. It was observed that triple improve kidney function following injury. However, poor survival negative cases showed a greater expression of gene as compared and grafting of stem cells to the site of injury has restricted their to other groups (p-value¼0.001). therapeutic efficacy. Preconditioning of stem cells can be a po- Conclusion: High expression of miR-21 in breast cancer pa- tential strategy; chemokines and growth factors can improve their tients suffering from stage III invasive ductal carcinoma had been potential in terms of homing and differentiation. Objectives of the calculated as compared to its age matched healthy subjects. This present study were to examine the changes in the expression result can mark miR-21 as a potentially strong diagnostic and profile of genes associated with renal injury and to explore the prognostic biomarker of breast cancer. MiR-21 profile was also novel strategy of preconditioning of MSCs for injured kidney cells. studied in daughters of the index cases. Their expression was also Methods: Rat AKI model was established with gentamicin and shown to be significantly higher as compared to the healthy in- renal specific genes were analyzed through RT-PCR. For pre- dividuals but lesser than the full blown disease of breast cancer. conditioning strategy we used IL-7 gene to transfect MSCs. MSCs This result strengthens the concept of inheritability of this disease were co-cultured with cisplatin-treated injured Mardin-Darby and this gene can also be labeled as a predictive biomarker. When bovine kidney (MDBK) cells and their in vitro fusion potential was Ct-values and fold change calculations were compared between analyzed. ER, PR and HER2, higher expression was seen in negative tumor Results: Several genetic changes occur in AKI that may con- tissues which are very resistant tumors and respond poorly to tribute in long term renal injury consequences and targeting these chemotherapy. pathways may appear to be effective therapeutic options. Flow- Serum metabolomic profiles for breast cancer diagnosis, cytometry of preconditioned MSCs and injured MDBK cells re- grading and staging by gas chromatography-mass spectrometry vealed significant (Pr0.001) cell fusion compared to that of the Naila I. Hadi1*, Qamar Jamal1, Ayesha Iqbal2, Fouzia Shaikh1, normal MSCs. In addition, we also observed improved migration Saleem Somroo3and Syed G. Musharraf2,4 ability of preconditioned MSCs in the in vitro wound healing assay. 1Department of Pathology, Ziauddin University, Karachi, Pakistan,2 Conclusions: For efficient stem cell therapy, it would be valu- Dr. Panjwani Center for Molecular Medicine and Drug Research, In- able to determine the contribution of various cellular factors in the ternational Center for Chemical and Biological Sciences, University of fusion process so that a rationale for the use of such precondi- Karachi, Karachi, Pakistan,3Breast Clinic, Surgical Ward 2, Jinnah tioned cells can be determined. Postgraduate Medical Center (JPMC), Karachi, Pakistan,4H.E.J. Re- Amino acid racemization in human dentine as an indicator search Institute of Chemistry, International Center for Chemical and of chronological age – A study in Karachi, Pakistan Biological Sciences, University of Karachi, Karachi, Pakistan. Qudsia Hassan1, Allah Rakha Aswad1, Zahid Bashir1and Tehreem Introduction and objectives: Detection of metabolic signature Ashraf1. for breast cancer (BC) has the potential to improve patient prog- 1Forensic Medicine Ziauddin University, Karachi, Pakistan. nosis. This study identified potentially significant metabolites Introduction and objectives: Amino acid racemisation is a differentiating between breast cancer patients and healthy con- reliable method to estimate age in developed countries. This study trols to help in diagnosis, grading, staging and determination of was designed to determine the coefficient of amino acid racemi- neoadjuvant status. sation (AAR) with reference to age in our population. The objec- Methods: Serum was collected from 152 pre-operative breast tives are to determine the coefficient of aspartic acid racemisation cancer (BC) patients and 155 healthy controls in this case-control (AAR) with reference to age by HPLC, to measure the mean error in study. Gas chromatography-mass spectrometry (GC-MS) was used calculating age using this method; and to identify any differences to obtain metabolic profiles followed by chemometric analysis. from earlier observation and discuss the reproducibility in our Results: Chemometric analysis showed the identification of setting. significantly differentiated metabolites including 7 for diagnosis, Methods: Hundred (100) teeth were obtained from subjects 19 for grading, 23 for staging, 15 for stage III subcategories and 10 between the age of 11 and 70 years who had come for routine for neoadjuvant status (p valueo0.05). Partial Least Square Dis- extraction procedure or for orthodontic procedures. Incisors, ca- criminant Analysis model revealed a distinct separation between nines, premolars and, first and second molars were included. healthy controls and BC patients with a sensitivity of 80% and Carious tooth and third molar were excluded. Samples were specificity of 100% on external validation. Models for grading, stored, dried, de-mineralized, hydrolyzed, and derivatised. High- staging and neoadjuvant status were built with Decision Tree Al- Pressure Liquid Chromatography (HPLC) was performed to quan- gorithm with predictive accuracy of 71.5%, 71.3% and 79.8% tify the L- and D-forms of aspartic acid in dentin. Correlation and respectively. regression was then tabulated based on the quantification, and Conclusion: Pathway analysis revealed increased glycolysis, was compared with data from other studies. lipogenesis, and production of volatile organic metabolites in- Results: It was observed that there is very strong positive dicating the metabolic alterations in breast cancer. correlation (0.818) observed between actual age and co-efficient of Growth factors mediated preconditioning of mesenchymal racemization (statistically significant at 5%), which indicates that stem cells: Strategy for enhancing renal tissue regeneration as age increase co-efficient of racemization increases. Based on the Rakhshinda Habib1,2, Kanwal Haneef1, Nadia Naeem1, Nurul analysis of 85 samples to estimate age least Square method was Kabir1, Atta-ur-Rahman1and Asmat Salim1. used to derived the regression line i.e. Age¼4.391þ347.396 (co- 1Dr. Panjwani Center for Molecular Medicine and Drug Research, efficient of Racemization). Co-efficient of determination was found International Center for Chemical and Biological Sciences, University to be 0.74, which show that regression equation for estimating age of Karachi, Karachi, Pakistan, 2Dow Research Institute of Biotechnol- was 74% correct. The computed regression line was tested by using ogy and Biomedical Sciences, Dow University of Health Sciences, coefficient of racemisation for 15 samples, we observed that the Karachi, Pakistan. mean of actual age (38.44713.22) was not significantly different 178 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 from the mean of calibrated age (37.52712.36) which is estimated supernatant. We also sought to further delineate the underlying by regression equation (P-value¼0.222 ). The mean error (differ- mechanisms elicited by oxidative stress and inflammation and ence between actual and calibrated age) is found to be 0.91177.41. suppressed by compounds. Conclusions: The correlation coefficient was strongly positive. Results: The results show that new Acylphloroglucinols deri- The results strongly suggests that aspartic acid racemisation of vatives and Myricetin significantly suppressed the oxidative stress human dentine is a precise method for estimation of chronological and anti-inflammatory efficacy in suppressing, total reactive spe- age in living and dead. The methodology should be standardized cies (ROS), superoxide, nitric oxide, pro-inflammatory inducible to make the results more accurate and prevent fallacies. However, nitric oxide synthase (iNOS), proliferation of T-cells and nuclear it remains an expensive procedure, considering that the laboratory factor-kappa B (NF-κB) nuclear translocation. All the compounds equipment is not available in most cities of Pakistan. were also non toxic to other cell systems. Correlating vitamin d deficiency with volumetric breast Conclusion: Based on these results, we conclude that natural density via a fully automated soft ware Volpara™ in the re- product may contribute in anti-inflammatory drug discovery. productive age group Imipenem resistant pseudomonas aeruginosa: the fall of the Bushra Wasim1, Khalid Khan1and Farah Habib1 final quarterback 1Department of Anatomy, Ziauddin University, Karachi, Pakistan. Nadya Ameen1, Zahida Memon1, Shehla Shaheen1, Ghulam Objective: To see the correlation of volumetric breast density Fatima1and Farah Ahmed1 and Vitamin D deficiency in the reproductive age group. 1Ziauddin University Karachi, Karachi, Pakistan Methods: We enrolled 300 patients from the general Introduction and objectives: To isolate, determine the fre- outpatient department, their blood samples were collected and quency, and study the demographic trends of MBL positive Pseu- vitamin D level was determined via chemiluminescence Method. domonas aeruginosa from imipenem resistant isolates collected Full field digital mammography was done and the DICOM images from clinical samples in a tertiary care hospital of Pakistan. were tagged and processed via fully automated software Volpara™ Methods: In this cross sectional study a total of 230 strains of by using the formula “Volumetric breast Density¼100 Volume of Pseudomonas were isolated from various clinical specimens on the fibroglandular tissue (cm3)/ Volume of breast tissue (cm3). basis of culture and biochemical tests. Imipenem resistant isolates Results: There is enough evidence now that Vitamin D defi- were selected by Kirby Bauer Diffusion technique, followed by ciency is inversely related to mammographic density and MD itself screening for MBL production by Imipenem EDTA Combined Disc is a strong predictor of breast cancer risk. Measurement of volu- Test. Demographic details of each patient were recorded on a se- metric breast density objectively via a fully automated software parate questionnaire. Chi-Square goodness-of-fit test was com- “Volpara" and using the Raw Processed version of the FFDM in this puted to review the isolation of MBL positive isolates (P-value study was attempted for the first time in Pakistan and correlating r0.05) in different specimen. with the vitamin D deficiency and also BI-RADS (Breast-Imaging Results: Out of 230 strains of P. aeruginosa 49.5% were imipe- reporting and Data system) which is a common method used in nem resistant; MBL production was confirmed in 64.9% of the clinical practice worldwide. The total number of patients had only resistant isolates. Resistance to polymyxin B (12.5%) was notable. two women with normal vitamin D level, the deficient group was Majority of the MBL positive strains were isolated from patients evaluated. In the group of women with low Vitamin D (Vitamin D aged between 20 and 39 years (45.9%) and the predominant o50), 76% had elevated VBD and 24% had normal VBD. The test for source was pus (43.24%) which was found to be statistically sig- proportions compares whether these two groups are significantly nificant (P-value¼0.04). Outpatient departments (24.3%) and burn different in size or not. unit (21.6%) were the major places for resistant isolates. Conclusions: The null hypothesis assumes that vitamin D de- Conclusions: MBL production is one of the major causes of ficient women have the same number of people in tolerable and IRPA. Increasing resistance to polymyxin B is grave. Due to ac- excess groups. Because the p-value is small, the null hypothesis is quisition of MBL strains, MDR P. aeruginosa has become endemic rejected and we conclude that the number of women with ele- in tertiary setups. vated breast density is not the same as the number of women with Antibiotic resistance in neonatal sepsis – study on low birth normal breast density, it is higher. cases from Karachi-Pakistan Natural compounds suppress oxidative stress and in- Mirza Tasawer Baig1*, Ali Akbar Sial1, Zahida Memon2and Nusrat flammatory response in activated mouse macrophages Bano3 Samreen Soomro1,3, M Ahmed Mesaik1, Farzana Shaheen2, Zaheer 1Department of Practice, of Pharmacy, Ziauddin Ul-Haq1and M Iqbal Chaudhary1,2 University Karachi,2Department of , Faculty of Phar- 1Dr. Panjwani Center for Molecular Medicine and Drug Research, macy, Ziauddin University Karachi,3Department of Pharmacology, University of Karachi, Karachi, Pakistan,2H.E.J. Research Institute of Faculty of Medicine, Ziauddin University, Karachi, Pakistan Chemistry, International Center for Chemical and Biological Sciences, Introduction and objectives: Premature neonates are often University of Karachi, Karachi, Pakistan,3Dow Research Institute of immunocompromised and readily susceptible to infectious dis- and Biomedical Sciences DUHS, Karachi, Pakistan eases. They are deficient in antibodies whereas neonatal neu- Introduction: Inflammation is quite complex, initiated by trophils are immature and depleted (2 fold in circulatory pool). several factors includes, molecules ranges from bacteria to che- Neonatal anatomic barriers are susceptible and hence infection mical causing cellular trauma. Tissue injury induced by this trau- control poses significant challenge. The objective of this study is to ma results in the inflammatory mediators release by macro-pha- assess the common pathogens isolated in neonatal sepsis and ges, including reactive oxygen species (ROS) like superoxide anion specify the antibiotic resistance pattern. (O2-), hydrogen peroxide (H2O2), nitric oxide and increased ex- Methods: The study was conducted in the neonatal ICU of two pression of pro-inflammatory cytokines, NF kappa B, and inducible public sector tertiary care hospitals inclusive of 234 cases. Pre- Nitric Oxide Synthase. A current study has led to the isolation of mature neonates with clinically confirmed bacterial infection were two new Acylphloroglucinols derivatives along with four known included comprising of 48.6% cases of Early onset neonatal sepsis compounds, Myricetin, Isousnic acid, G3-factor and myrto- (EONS) with obstetrical complication and fulminant, multisystem commulone E from Myrthus. disease. Intrapertum fever was recorded in 87% cases of EONS Methods: In this study, we used Lipopolysaccharide induced among which, 22% culture positive neonates were initially macrophages to study immune mediators release in culture asymptomatic. Apnea, fever and metabolic acidosis were the more N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 179 prevalent symptoms. CBC, C-reative protein, Glucose and Blood concentrations of these compounds were added separately into culture test were recorded. the culture medium and the treated cells were analyzed for the Results: The most common pathogen was Group B streptococci expression of cardiac specific genes by RT-PCR and cardiac specific (24%), Klebsiella pneumonia (21%) and E.coli (15%). Fetal hypoxia proteins by immunocyto-chemistry and flow cytometry. The was confirmed in 34% cases. Ampicillin and Gentamycin was given treated MSCs were then transplanted to the rat model of myo- empirically in 87% cases. Modified Kirby Baur disc diffusion tech- cardial infarction and cardiac functional studies were performed nique was used to determine antibiotic resistance. Klabsiella by echocardiography. pneumonia was greatly resistant against cephalosporins (average Results: We have found expressions of cardiac specific proteins of 54%) and co-trimoxazole (87.9%) whereas comparatively sus- and genes in the modified mesenchymal cells both at genetic and ceptible to quinolones (ciprofloxacin). Resistance of amikacin was protein levels. Transplantation of modified MSCs into infarcted elucidated in 87% cases. myocardium of rat improved the cardiac left ventricular function Conclusion: In lieu with preventive approaches to avoid neo- possibly by enhancing the rate of bone marrow stem cell differ- natal sepsis, careful treatment choice of antibiotic is required to entiation into mature cardiomyocytes in the infarcted heart. deplete chances of therapeutic failure. Conclusions: We therefore, conclude that pre-treating auto- Withania coagulans: A potential cholesterol lowering plant logous mesenchymal stem cells before transplantation may in- Tooba Lateef1, Sadaf Naeem1and Shamim A Qureshi1 crease the likelihood of successful regeneration of damaged 1Department of Biochemistry, University of Karachi, Karachi, myocardium. The study put forth another valuable aspect that Pakistan would serve as a tool for modified cellular therapy. Introduction and objective: Withania coagulans has many Antiplatelet aggregating and membrane stabilizing proper- medicinal properties. Currently its cholesterol lowering effect was ties of areca nut investigated in terms of HMG-CoA reductase inhibiting activity. Shagufta Khan1and Ahsana Dar2 Methods: The study was designed to investigate in-vivo effect 1Department of Biological and Biomedical Sciences, Aga Khan of methanolic fruits extract (MFEt) of W. coagulans in hyperlipi- University Hospital, Medical College, Karachi, Pakistan,2HEJ Research demic rabbits. On the completion of trial, total cholesterol (TC) and Institute of Chemistry, ICCBS, PCMD, University of Karachi, Karachi, HMG-CoA/mevalonate ratio were estimated. Additionally, in-silico Pakistan study was performed to find out the inhibitory effect of 21 che- Introduction and objectives: Platelet aggregation inhibitors mical compounds present in fruits of same plant on HMG-CoA are one of the key preventive and therapeutic agents for cardio- reductase (HMGCR). Molecular docking calculations were carried vascular diseases. Areca nut is used in folk medicine for treatment out by using Molegro Virtual Docker (MVD) software. of various ailments. Present study was conducted to investigate Result: All doses of MFEt were significantly (po0.05) de- the antiplatelet aggregating and membrane stabilizing activities of creased TC and improved HMG-CoA/mevalonate ratio indicating areca nut extract and its fractions using platelet aggregation and HMGCR inhibition. Interestingly, 5 compounds were displayed the membrane stabilization assays. highest MolDock score including Ergosta-5, 25-diene-3 β,24ε-diol, Methods: To determine the effect of areca extract and its Withacoagulin, Withanolide D, Coagulin D and Withaferin. Of fractions on platelet aggregation, blood was taken from healthy which, excluding Withanolide D rest of the 4 compounds inter- human volunteers and platelet aggregation was monitored. acted with catalytic residues present in the active site of HMGCR Membrane stabilizing properties were assessed using human red by forming hydrogen bonds and inhibited the activity of same blood cells. enzyme. Results: Our results showed that areca extract was effective Conclusions: The present in-vivo and in-silico studies proved against arachidonic acid induced platelet aggregation in dose de- that compounds present in fruits of W. coagulans are predicted as pendent manner leading to complete inhibition at 10 mg/ml dose. potent inhibitors of HMGCR that could be used for the develop- Data obtained from the present study showed that the areca ex- ment of drug for hypercholesterolemia. tract (EC50¼91 mg/ml) and its aqueous fraction (EC50¼55 mg/ml) Cardioprotective effect of modified bone marrow mesenchy- possess significant membrane stabilizing activity in a dose de- mal stem cells after transplantation in rat model of myocardial pendent manner (IC50¼100 mg/ml). infarction: An applied approach towards cellular therapy Conclusions: These findings suggest that areca nut extract, Nadia Naeem1, Muhammad Aleem Akhter1, Irfan Khan1, Hana’a ethylacetate fraction and aqueous fraction possess significant an- Iqbal1, Kanwal Haneef1, Nurul Kabir1and Asmat Salim1 tiplatelet aggregating and membrane stabilizing properties, and 1Dr. Panjwani Center for Molecular Medicine Drug Research, In- aqueous fraction was most effective which may be of clinical value. ternational Center for Chemical and Biological Sciences, University of However, further studies are required to identify its active com- Karachi, Karachi, Pakistan pound(s) via bioassay guided-fractionation method. Introduction and objectives: Myocardial infarction is in- Small molecular activators of proteasome-related HSLV creasing cause of death worldwide. In , cardiac diseases peptidase are increasing at a rate greater than in other part of the world. Yasmeen Rashid1 Regenerative medicine involves the repair or regeneration of an 1Department of Biochemistry, University of Karachi, Karachi, organ, tissue or cells in order to restore an impaired function of the Pakistan tissue. The cellular therapy offers a promising therapeutic strategy Introduction and objectives: The HslVU or prokaryotic mini- for the treatment of different diseases including myocardial in- proteasome is two component system based on HslV peptidase farction. Mesenchymal stem cells (MSCs) are having immense and HslU ATPase. Like proteasome it is involved in intracellular importance in the emerging field of regenerative medicine and proteins degradation. Due to its occurrence in pathogenic mi- have tremendous capability for cellular therapy and tissue en- crobes and absence in human beings, it is considered as a potential gineering. MSCs can be converted into cardiomyocytes by treating antimicrobial drug target. The functional HslVU complex forms them with DNA demethylating agents. Our aim is to investigate the when HslV dodecamer is flanked at both sides by HslU hexamers cardioprotective effect of pretreated MSCs when transplanted in and the carboxy termini of HslU subunits intercalate into a clefts infarcted myocardium of rat. between two adjacent HslV subunits. This intercalation is followed Methods: In the present study, rat mesenchymal stem cells by conformational change in HslV active site resulting in its al- were treated with DNA-demethylating agents. The optimized losteric activation. We for the first time report certain small 180 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 molecules which are capable of activating HslV peptidase in the University, Karachi, Pakistan,2Department of Pathology, College of absence of its natural activator, HslU. Medicine and Health Sciences, UAE University, UAE The benzimidazole, quinazoline and chromone derivatives were Introduction and Objectives: Myocardial infarction (MI) leads suggested by ligand docking studies to bind at the carboxy terminus to complex repair processes including scarring and fibrosis. Ga- binding pocket of HslV in a manner similar to HslU carboxy termi- lectin-3 (Gal-3) is a beta-galactoside binding lectin responsible for nus. This was confirmed by HslV activation assays with these com- cardiac fibrogenesis and adverse left ventricular (LV) remodeling. pounds that gave ED50 in micromolar range (0.6–1.5 mM). Canonical Wnt signaling in myofibroblasts following MI also Results: The results showed that small, extracellular non- modulate wound healing and fibrosis. As Gal-3 and Wnt 3a play a peptidic molecules can allosterically activate the peptide hydro- significant role in the post MI sequel we aim to measure their lytic activity of HslV, which in turn would initiate intracellular concentration in murine heart at 7 week post Ischemia/reperfu- proteolysis. sion injury and to determine their correlation. Sensitivity pattern of Moraxella catarrhalis isolated from Methods: Male C57B6/J mice were used for myocardial IR in- hospitalized patients jury by temporarily ligating the left anterior descending artery of Naheed A.Irfan1, Maha Jamil1, Aziz Fatima1and Syedda Gufrana the heart for 30 min to create ischemia/infarction followed by Nadeem1 restoration of blood flow. Samples collected 1 week post IR injury 1Jinnah University for Women, Karachi, Pakistan point was processed for ELISA to detect Gal-3 and Wnt-3a levels in Introduction and objectives: Moraxella catarrhalis (formerly the heart. known as Branhamella catarrhalis) has emerged as a significant Results: Gal-3 and Wnt 3a levels were significantly increased in bacterial pathogen of humans over the past two decades. Over the the LV heart tissue at 1 week post IR compared to sham operated same period, studies have revealed its involvement in respiratory mice. There was a strong and significant positive correlation ob- (e.g., sinusitis, otitis media, bronchitis, and pneumonia) and ocular served between Gal-3 and Wnt-3a at 1week post IR injury. infections in children and in laryngitis, bronchitis, and pneumonia Conclusions: Gal-3 and Wnt-3a levels were increased in in adults. It has discovered an increasing number of Β-lactamase- 1 week post myocardial IR injury in conjunction with intrinsic positive strains. We aim to find out the sensitivity pattern of cardiac repair. Understanding the mechanisms of this repair pro- Moraxella catarrhalis isolated from sputum. cess could lead to optimal and prevention or delay in Methods: Total 124 sputum samples were analyzed. Con- the onset of heart failure. firmation done by using chocolate agar a typical golden yellow STD pathogens determined in semen using PCR and “flow- which can be distinguished by hockey puck like colony. Further through” hybridization technology identifications were done by gram staining, oxidase test and bio- Rubina Ghani1 chemical test. Antibiogram was done by CLSI method. Statistical 1Department of Biochemistry, Pathological and Molecular La- analysis was also performed using SPSS version 17. boratories, Department of , Jinnah Medical and Dental Results: Sensitivity pattern showed AMC 99%, TE 49%, CRO 01%, College, Karachi, Pakistan E 30%, SXT 04%, CIP 50%, CEF 84%. Background: The prevalence of sexually transmitted Disease Conclusions: To control the resistivity of these organisms some (STDs) in health care facilities for diagnosis of common pathogens, preventive measurements like proper identification and selective those causing infertility like Chlamydia trachomatis, Neisseria should be taken. gonorrhoeae and Mycoplasma hominis. Genital wart is a highly Assessment of HCV genotypes gene flow hybridization and contagious sexually transmitted disease caused by some sub-types its comparsion with conventional PCR of human papillomavirus (HPV). Nazish Jaffar1, Rubina Ghani2and Saba Sattar1 Methods: Semen samples were obtained by masturbation into 1Jinnah Medical University, Karachi, Pakistan,2Pathological sterile containers after sexual abstinence of 48–72 h. Samples were and Molecular Laboratories, Karachi, Pakistan subjected to semen analysis within one hour of collection. The Background: Nucleotide sequence analysis of hepatitis C virus concentrations of sperm as well as sperm motility were also de- (HCV) strains showed substantial variability leading to a classifi- termined. DNA extraction was extracted of all the samples and the cation into several genotypes and subtypes. PCR assay was performed. The amplicons are subsequently hy- Objective: The objectives of the present study were the bridized to pathogen-specific capturing probes via “Flow-through” knowledge of the HCV genotypes in Pakistan, and determining hybridization. their prevalence. Result: During our study we came across with the STI patho- Methods: During the course of this study, HCV-RNA positive gens present in our population and the reason for infertility was sera samples from 200 chronically infected patients were char- the main cause. When Chlamydia trachomatis and Neisseria go- acterized by genotyping assay and Automated Sequence Analysis. norrhoeae were detected in their wife’s were also screened and Genotyping assay utilized type-specific primers for amplification these STI pathogens were identified. of the core region, where as sequencing was done for 5′Noncoding Conclusions: The main route for the transfer of STI pathogens region of HCV using ABI 3100 Sequencing Analyzer. were the men special those who visited commercial sex workers Results: During our study we came across that genotype 3a as they were working in other cities and the complained for in- was the most prevalent (36%) followed with 3b (26%), 3c (3.5%), 3d fertility. Screening for bacterial STI pathogens, Mycoplasma ho- (4%), 1a (2.5%), 1b (1%), 1c (2.5%), 1d (0.5%), 2a (12%), 2b (0.5%), 2c minis, Chlamydia trachomatis and Neisseria gonorrhoeae are (0.5%), 4 (0.5%) and 4a (1.5%). Genotypes 3d, 1c, 1d and 2c were strongly recommended because these pathogens can cause serious reported for the first time from Pakistan. reproductive complications. Conclusion: Advances in the field of molecular biology have Effect of Citrus paradisi on coagulation mechanism provided rapid diagnostic tools that have reduced the turnaround Neelam Mallick1, Ali Akbar Sial1and Huma Ali1 times for detecting HCV genotype by using “Flow-through” hy- 1Faculty of Pharmacy, Ziauddin University, Karachi, Pakistan bridization in Pakistan. Introduction and Objectives: Over the past few years, herbal Galectin-3 and Wnt signaling in myocardial infarct healing: remedies and medicinal plant entities are of great interest in Is there a connection? various region of the world for their therapeutic and preventive Satwat Hashmi1 and Suhail Al-Salam2 effects in various cardiovascular ailments like atherosclerosis. 1Department of Biological and Biomedical Sciences, Aga Khan Numerous animal models were studied extensively for elucidation N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 181 of these plant component effects on different organs. Citrus resistant to therapy. There has been very little, if any, work done paradisi (C. paradisi ) due to its antioxidant and anti-inflammatory on the precise contribution of cell surface muscarinic GPCRs in potential has been selected in this study to evaluate the effect of prostate cancer growth. this compound on coagulation and anticoagulation factors. C. Methods: Here we determined the role of the M1 receptor in paradisi contains high concentrations of hesperidin, naringin, li- prostate cancer cell proliferation. The treatment of PC-3 cells with monene and related flavonoids. selective M1 receptor antagonists, Dicyclomine exhibited anti- Methods: Doses were selected in the range of 0.1–0.5 ml/kg. proliferative effects using growth assays, while M1 receptor ago- Aspirin and warfarin 150 mg/kg and 5 mg/kg-10 mg/kg respec- nist, Pilocarpine exhibited increased proliferation compared with tively were used as standard anticoagulants. Animals were divided the untreated cells. into six groups with ten rabbits in each group. Results and Conclusions: Further studies will dissect mole- Results: At median dose level, significant enhancement in cular pathways to determine the role of the M1 receptor in an- thrombin, pro-thrombin time was observed in comparison to drogen sensitive and androgen insensitive prostate cancer cell control, while fibrinogen levels were significantly reduced with growth and proliferation. The results of this work may help inform warfarin. Furthermore, substantial inhibition of platelet aggrega- clinical and public health interventions for prostate cancer by tion by adenosine phosphate, collagen, epinephrine and arachi- providing insights into the biology of prostate cancer progression. donic acid was observed at median C. paradise dose (MCPD). Co and contra-regulation of glycosylated proteins in three Conclusions: Results of this study have shown an effective distinct regions of schizophrenics brain anticoagulant activity. Thus the patients, who are at risk of car- Ayesha Khan1, Nikhat Ahmed2, Beena Hasan1and Shamim diovascular event development may additionally be advised to Mushtaq2 consume grape fruit juices in their diet to reduce the chances of 1Neurochemistry Research Laboratory, Department of Biochem- mortality and associated morbidities. istry, University of Karachi, Karachi, Pakistan,2Department of Bio- Conventional clinical and prognostic variables in 150 oral chemistry, Ziauddin University, Karachi, Pakistan squamous cell carcinoma cases from the indigenous popula- Introduction and objectives: Schizophrenia, a multifactorial tion of karachi disorder exhibits diverse neuropathological aberrations with al- Mohiuddin Alamgir1 tered protein expression and post translational modifications as a 1Bharia University, Karachi, Pakistan surfacing evidence that may also contribute to its pathophysiology. Objectives: To evaluate conventional clinical and prognostic To investigate the integrated picture of molecular changes and variables in Oral Squamous Cell Carcinoma (OSCC) cases from the potential alterations in expression of specific glycosylated proteins indigenous population of Karachi. in different brain regions compared to physiologically normal Methods: Common clinical and pathological parameters re- brains. We aimed to provide more holistic view of three brain lated to poor prognosis of 150 OSCC cases presented at two major regions; and their dynamic cross-talk providing insights into the treatment facilities of Karachi were evaluated. The reporting in- underlying molecular mechanisms associated with schizophrenia. cluded demographic details and variables like intra-oral subsites, Methods: Present study entails, differential proteomic analysis clinical stage and histological grades. Recurrence of tumor after of autopsied brain regions of schizophrenic; substantia nigra, initial resection was also documented. cortex, and hippocampus (n¼7 each), by using sodium dodecyl Results: The patient’s population comprised of 98 males and 52 sulphate polyacrylamide gel electrophoresis coupled with im- females. The mean age was 47.1þ12.22 (range: 20–78 years). munoblot and DIG (digoxigenin) labelling followed by ESI-QTOF Maximum numbers were seen in the 41–50 years age group. - MS analysis for validation. speaking community was the most affected ethnic group (n¼75). Results: We have identified fourteen glycosylated protein Histopathological analysis revealed that majority of cases was components with altered expression among the brain regions. The moderately differentiated tumors (59%) with clinical stage II (35%) 50KDa (Glial fibrillary acidic protein, GFAP) and 84KDa (mi- or IV (29%). The most common intra-oral subsite came out to be tochondrial inner membrane protein) are contra-regulated be- buccal mucosa of cheeks (56%) followed by lateral borders of tween substantia nigra and cortex. While T-complex protein tongue (21%), lips (13%), alveolar (6%), palate (2.6%) floor of mouth 1 subunit zeta of 58KDa is co-regulated between substantia nigra (1.3%), etc. Recurrence was observed in 8 out of 150 cases. All and hippocampus. The remaining five proteins identified with patients underwent primary resection7neck dissection and re- significantly altered glycosylation intensity in the specific brain construction where possible. regions give evidence of their explicit regional function. Ad- Conclusions: Overall experience with oral squamous cell car- ditionally, nine proteins commonly appeared non-glycosylated in cinoma shows that it has a high tendency for local invasion as well the three studied brain regions. as dissemination to regional lymph nodes, i.e. cervical lymph Expression and association of CDK10 with ETS2 during hu- nodes, both are associated with a poor prognosis. Preventable risk man corneal wound healing factor of tobacco chewing has been observed in majority of these Meraj Zehra1, Shamim Mushtaq2and Nikhat Ahmed Siddiqui2. cases. 1Department of Biochemistry, Karachi University, Pakistan,2De- Poster Presentations. partment of Biochemistry, Ziauddin University, Karachi, Pakistan. Role of muscarinic G-protein coupled receptor (GPCR) in Introduction and objectives: Corneal related complications prostate cancer growth and survival are major health concerns worldwide because its progression is Abdul Mannan Baig1and Farhat Abbas1 associated with significant impaired vision. Therefore, there is an 1Department of Biological and Biomedical Sciences, Department of urgent need to develop reliable understanding of the underlying Pathology and Microbiology, Department of Surgery, Aga Khan Uni- mechanism of corneal epithelial wound healing to apply ther- versity, Karachi, Pakistan apeutic options. We aimed to investigate the alterations in protein Introduction: Despite advances in early detection and treat- expression and association of CDK10 and ETS2 during corneal ment, prostate cancer remains one of the most frequently diag- epithelial migration and to demonstrate the networks of the total nosed cancers and a leading cause of cancer death in males identified proteins with potential dual functions. worldwide. Much work has been done on androgen receptors in Methods: In this study, human corneal epithelial cells lines relation to proliferation and survival, and it has been shown that (HCEC) have been used for wound healing model. Mechanical cancers of this organ usually turn androgen insensitive and wound was made in HCEC lines and healing was monitored at 24, 182 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

48 and 72 h of post wounding. Epithelium was scrapped at 24, 48 Aaida Mumtaz1, Aqsa Khalid1, Zahir Hussain2, Faiza Alam1and and 72 h, followed by protein quantification using BCA kit. The Rehana Rehman1 wounded and unwounded cells were subjected to SDS-PAGE and 1Aga Khan University, Karachi, Pakistan,2Umm Al-Qura University, two-dimensional electrophoresis (2DE). Mass Spectrometry Kingdom of Saudi Arabia (MALDI TOF) was done to identify the proteins through protein Introduction and Objectives: To compare Kisspeptin (KP) le- database searches. The identified protein were further analyzed vels in females with infertility to male, female and unexplained and validated by western blot analysis. A further insight into the causes and associate it with success of treatment after ICSI. links among the identified proteins and their functional roles were Methods: Cross sectional study carried from August 2014 till analyzed by STRING 8.3 software. May 2015 included all categories of infertility with duration 4 Results: A significant finding of the present study is the iden- two years, female age between 20 and 50 years and body mass tification of Cdk10, EFNB3, RAB 34, RRAS, HSP2 and HSP90 in index 418 o35 kg/m2. Down-regulation of ovaries was followed healing corneal epithelium at active phase of migration. Expres- by calculated stimulation, ovulation induction, oocytes retrieval, sion of CDK10 and ETS2 were validated using antibody by western ICSI, in vitro maturation of embryos and finally blastocysts trans- blot. Interaction association network analysis further confirms the fer. KP levels were measured by enzyme linked immuno sorbent close interacting relationship between CDK10 and ETS2 proteins. assay on day of ovulation induction. Failure of procedure was de- Conclusions: These findings are one step forward in identifying tected by beta human chorionic gonadotropin o5-25mIU/ml the mechanism of wound repair or re-epithelialization. Associa- (non-pregnant) whereas females with levels 425mIU/ml and no tion of CDK10 and ETS2 in migrating samples suggested that they cardiac activity had preclinical abortions. Clinical pregnancy group have positively regulated the cell cycle and cell proliferation and had confirmed cardiac activity on trans vaginal scan. Data was hence mediate the wound healing. This study may also increase analyzed using SPSS 15. the understanding of normal and abnormal corneal function with Results: Females who completed the procedure (168) were likely relevance to corneal disease and transplants. stratified; 40 (24%), 66 (39%) and 62 (37%) with male, female and Functional association of bad and 14-3-3 proteins in schi- unexplained infertility. The KP levels were 285.38723.96 ng/L zophrenic patients (mean7SD). The females with unexplained infertility had lowest Sobia Manzoor1, Shamim Mushtaq2, Ayesha Khan1and Nikhat KP (101.4711.96) which was statistically significant (po0.05). Ahmed2 The results declared 66 (39%) females as non-pregnant, 30 (18%) 1Department of Biochemistry, University of Karachi, Pakistan,2 with preclinical abortions and 72 (43%) with clinical pregnancies. Department of Biochemistry, Ziauddin Medical University, Karachi, Out of non-pregnant females 42 (64%) had unexplained cause of Pakistan infertility. Introduction and objectives: Schizophrenia is a multifactorial Conclusions: Low KP levels in unexplained females explains disease which involves the exogenous and endogenous interaction the peripheral role of the neuropeptide, deficiency of e¼which and is characterized by neuronal reduction, dendrite deficits, de- leads to failure of implantation after ICSI. creased synaptic markers and alteration in synaptic circuitry. Reliability of nephrin as an early diagnostic biomarker for Schizophrenia is a neurodevelopmental disorder but apoptosis screening diabetic nephropathy may contribute to the pathophysiology of schizophrenia due to Moazzam A Shahid1, Saeeda Baig1, Adnan M Zubairi1and Farha progressive clinical deterioration and subtle neurostructural Ahmed1 changes following the onset of psychosis. Apoptosis is a mechan- 1Ziauddin University, Karachi, Pakistan ism of cell death, is regulated by a complex cascade of pro and Introduction and objectives: Diabetic nephropathy occurs as a anti-apoptotic proteins. Dysregulation of apoptosis in several cor- result of proximal tubule dysfunction with podocyte damage due tical regions of schizophrenia indicates that apoptotic vulnerability to increased advanced glycation end-products insults in diabetes. is increased. Pro-apoptotic protein BAD (Bcl-2 associated death Nephrin, one of the three proteins that make up the podocyte promoter) interacts with anti-apoptotic BCL-2 and inhibit its ac- architecture, is excreted foremost following renal damage. The aim tivity while 14-3–3s binds with BAD and inhibit apoptosis that it of this study was to evaluate the efficiency and reliability of ne- act as an ant apoptotic protein. Various proteomic based studies phrin for an early biomarker of kidney damage in diabetic patients. shown reduction of 14-3-3 proteins in schizophrenic brains. In this Methods: Urine samples (78) were collected from diabetic study we aimed to identify and validate the differential expression center. Protein and glucose were determined by Dipstick. The of BAD and 14-3–3s by western blotting and their association patients were grouped on the basis of Albumin/creatinine ratio network with other proteins by String 10, Qiagen pathway data (UACR) as normoalbuminuric, microalbuminuric and macro- bases. albuminuric with UACR less than 30 mg/g, from 30 to 300 mg/g, Methods: Expression of BAD and 14-3-3 proteins from autop- and above 300 mg/g respectively. ELISA (Exocell USA) was used for sied schizophrenic brain cortex were validated by western blotting Nephrin estimation. Statistical evaluation was done. and In Silico analysis was done to see the association with other Results: Nephrinuria was present in 70 (89.7%) out of 78 dia- proteins by using String database, Qiagen pathway. betic patients including 35 (81.4%) normoalbuminurics, 5 (6.4%) Results: We have identified differential expression of BAD and micro-albuminurics and all 5 of macroalbuminurics (po0.027). 14-3-3 proteins in cortex region of schizophrenic human brain by When associated with duration of diabetes, 21 (91.3%) positive western blotting. Interaction association network between BAD with less than three years of diabetes, 24 (92.3%) with three to and 14-3-3 (YWHAQ, YWHAH, YWHAE, YWHAB, YWHAZ,Y- seven years and 25 (86.2%) in more than seven years (po0.039). WHAG) was analyzed by string software. Qiagen pathway further Nephrin levels were found increasing from Normo-(0.86 μg/ml) to confirms the close interacting relationship between BAD and 14-3- Micro-(11.6 mg/ml) to Macroalbuminuria group (47.6 mg/ml), com- 3 proteins in apoptotic pathway related to schizophrenic patients. pared to 0.15 mg/ml in comparison group. Conclusions: Apoptotic proteins BAD and 14-3–3s association Conclusions: The increase in nephrin levels from 0.86 mg/ml in in Schizophrenic brain cortex region provide insight into the patients with normal albuminuria to 47.6 mg/ml in patients with apoptotic pathways which play role in the pathophysiology of macroalbuminuria suggests that Nephrin precedes albumin in schizophrenia which could lead to development of therapeutic urine predicting early signs of kidney damage. drugs to target apoptotic proteins and their interacting partners. Bioinformatics navigation predicted structure-function dy- Kisspeptin and unexplained infertility namics of Calpain10 isoforms N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 183

Farina Hanif1, Bushra Chaudhry1and Kausar Saboohi1 Methods: Aortae from Sprague-Dawley rats and atria from 1Department of Biological and Biomedical Sciences, The Aga Khan guinea-pigs were placed in 5 ml and 15 ml isolated tissue bath University Medical College, Karachi, Pakistan assemblies respectively, which were maintained at 37 °C with Introduction and objectives: Calpain10, a member of cysteine constant carbogen supply, filled with physiological salt solution proteinase family, is present in eight different isoforms (a-h) exact (Kreb’s solution) and connected to a force transducer and Power- mechanism through which it influences the insulin secretion and Lab attached with a computer. action is scanty. Results: Bl Cr, in isolated guinea-pig atria, equally inhibited Methods: We have applied bioinformatics approach to en- force and rate of spontaneous atrial contractions. When tested on visage its mechanism of action and possible involvement in dis- phenylephrine (PE, 1 mM) and Kþ(80 mM)-induced vasoconstric- eases. National Center for Biotechnology Information Blast was tion, Bl Cr caused a concentration-dependent relaxation and also used for humans’ nucleotide and protein sequence alignments of caused a suppression of PE (1 mM) control peaks in Ca2þ-free the calpain10 isoforms a, c and g. Further, NCBI conserved domain medium. tool and CDART were used for other species. Conclusions: These data indicate that Bl Cr exhibits cardio- Results: Results of blast showed 100% nucleotide whereas 91% suppressant and vaso-dilatory properties. The vasodilator effect of and 97% amino acid sequence identity among isoforms of Homo the plant extract is mediated through inhibition of Caþþ influx sapiens’ calpain10 c and g with calpain10 a. There was 82% query via membranous Caþþ channels as well as Caþþ release from coverage of calpain10 c and 17% of calpain10 g with calpain10 a. intracellular stores. Further studies are required to elaborate the Conserved domain analyses of peptide sequences indicated do- anti-hypertensive activity of the plant. mains IIa and IIb have significant homology with the catalytic Association of Hepatitis G with liver dysfunction in treat- domain of the cysteine protease superfamily in all three isoforms. ment responders Hepatitis C patients The domain III and IV of isoform a and domain III of isoform c were Uzma Najam1, Syed Tousif Ahmed1and Rubina Ghani2 found homologous with linker C2 like subdomain III of mu- 1Department of Physiology, Ziauddin University, Karachi, calpain. Pakistan,2Department of Biochemistry, , Conclusion: It is predicted that calpain10 a and c isoforms may Karachi, Pakistan have a role in developing diseases such as neurodegenerative, Background: About 10 million Pakistani population is infected cardio-vascular, cataract, osteopenia and cancer. Whereas isoform with Hepatitis C virus (HCV). The prevalence is even more pro- g may exhibit protease activities having papa-in-like domain only. nounced among high risk population. Combination of standard TP53 protein overexpression in oral squamous cell carcino- interferon and ribavirin is still the first line therapy with sustained ma (OSCC): correlation with histologic variables and 5 years viral response (SVR) up to 40–50%. Pool of relapsers and non-re- survival in Pakistani patients sponders is increasing in Pakistan and needs treatment with pe- Adnan Ali1, Sohail Awan1, Omar Irfan2and Hamza Khan2 gylated interferon plus ribavirin (peg IFN/RIB). The hepatitis G 1Aga Khan University Hospital, Karachi, Pakistan,2Aga Khan Uni- virus (HGV), a recently identified member of the Flaviviridae fa- versity Medical College Karachi, Pakistan mily, can cause chronic infection in man. Introduction and objectives: Alteration of TP53 gene in oral Methods: 250 treatment responders Hepatitis C patients (un- squamous cell carcinoma (OSCC) is believed to be associated with detectable HCV RNA in the serum after 24 weeks of post treatment reduced overall survival (OS), the 5 year prognosis in our study follow up) were recruited from Jinnah Post Medical College and and the disease-free survival (DFS). The aim of this study is to Ziauddin Hospital. determine whether TP53 protein over expression in OSCC is a Liver function tests of all patients were assessed by automatic prognostic indicator of survival in Pakistani cancer patients along analyzer kits. HGV RNA was investigated in serum samples by with its correlation with risk factors including smoking, chewing reverse transcription and polymerase chain reaction amplification habits, histological variables like grade and stage of the tumor in a of the 5′ non-coding region of HCV and hybridization to a specific high risk population. probe. The liver function tests of HGV RNA sero-positive and sero- Methods: A total of 140 patients of OSCC were part of our negative patients were compared. study. TP53 protein over expression was investigated by means of Results: A total of 250 treatment responder hepatitis C patients immunohistochemistry. were evaluated with a mean age 4475.55. Males constituted Results Overexpression of p53 protein was observed in 75 38.4% (N¼96) while female constituted 61.6% (N¼154) of our patients (54%) using a threshold of 10% stained tumor nuclei. Pa- study group. Mean total billirubin, direct billirubin and indirect tients with p53 negative tumors had improved OS when compared billirubin was 0.77270.19 mg/dl, 0.30870.99 mg/dl and with patients with p53 positive tumors. This difference was sta- 0.49670.55 mg/dl respectively. Mean ALT was 81.97738.03 tistically significant (p¼0.036) in univariate Cox regression ana- units/L and total protein was g/dl 5.2570.44. Statically significant lysis however, it lost its worth in the multivariate analysis. difference of mean LFTs was found between hepatitis G positive Conclusion: This works supports that patients with p53 over- and negative patients with p-value ¼0.001. To find out association expression had a significantly poor overall survival compared to of LFTs with hepatitis G positive patients, chi-square was used. p53 negative patients. However, p53 overexpression was not as- Total billirubin, direct billirubin, indirect billirubin, AST, GGT, and sociated with patient’s disease free survival. APTT are found to be associated with hepatitis G with p-value of Cardiovascular effects of aqueous-methanolic extract of 0.001. By Log Regression we found significant and positive asso- bergenia ligulata in experimental animals ciation of Hepatitis G with Total billirubin, direct billirubin, in- Hasan Salman Siddiqi1and Mohammad Ahsan Sohail1 direct billirubin, GGT and APTT (p-value ¼0.001 at 95% CI). 1Department of Biological and Biomedical Sciences, Medical Col- Conclusions: Liver dysfunction in treatment responder hepa- lege, Aga Khan University, Karachi, Pakistan. titis C patients is found to be associated with novel Hepatitis G Introduction and objectives: Prevalence of hypertension, a virus. major risk factor for diseases like ischemic heart disease and cer- Combination therapy of non-steroidal anti-inflammatory ebrovascular stroke, is on the rise. The aim of this study was to drugs and anti-oxidants: An effective regimen for in- explore the cardiovascular effects of the aqueous-methanolic ex- flammatory disorders? tract of Bergenia Ligulata rhizome (Bl Cr), to provide pharmaco- Shagufta Khan1, Osama Anis Silat2and Ahsana Dar3 logical basis for its medicinal use in hypertension. 1Department of Biological and Biomedical Sciences, Aga Khan 184 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

University Hospital, Medical College, Karachi, Pakistan,2Ziauddin gender on it. University, Faculty of Medicine, Karachi, Pakistan,3HEJ Research In- Methods: 300 eyes from 150 healthy subjects aged 40 years stitute of Chemistry, ICCBS, PCMD, University of Karachi, Pakistan and above with no ocular pathologies were examined using Introduction and Objectives: Non-steroidal anti-inflammatory standard protocols by a single examiner. Subjects with of drugs are of therapeutic value for inflammatory disorders. Reactive diabetic or hypertensive retinopathy, raised intraocular pressure Oxygen Species may initiate and/or aggravate inflammation. In- (421 mmHg) and previous intraocular or laser surgery were ex- formation regarding the interaction of vitamin E with NSAIDs in cluded from the study. The mean retinal nerve fiber layer thickness inflammation is not well documented and is controversial. was calculated and was correlated with age and gender difference. Methods: This study was conducted to determine the effect of Results: The mean global retinal nerve fiber layer thickness was aspirin with vitamin E supplementation in inflammation, and to found to be 99.0279.08 mm in our set of population. Out of four explore the possible interactions of vitamin E with one drug from quadrants the maximum RNFL thickness was found in inferior each sub group of NSAIDs with respect to their anti-inflammatory quadrant (126.45716.23 mm) followed by the thickness of activity using animal model of inflammation, DNA degradation 121.50715.03 mm in superior quadrant, 70714 mm in nasal and lipid peroxidation assays. quadrant and 68.90713.10 mm in temporal quadrant. We found Results: Our results showed that vitamin E caused 10% in- strong negative correlation of RNFL thickness with age (Pr 0.001) hibition in edema at 400 mg/kg dose while aspirin elicited com- and not significant relation with gender (P¼0.8). plete inhibition in edema at 200 mg/kg dose. Co-administration of Conclusions: Keeping in mind the variations in RNFL thickness different doses of aspirin with vitamin E showed a dose-depen- with ethnic differences, this study provides the normal values of dent increase in edema inhibition. At 100 mg/kg aspirin showed RNFL thickness according to our set of population. It is concluded 50% inhibition in edema. While the same dose of aspirin with that RNFL thickness decreases significantly with increasing age but vitamin E supplementation (200 mg/kg) showed a significant in- gender had no significant effect on it. crease (25%) in its anti-inflammatory effect. Similar results were Prevalence of metabolic syndrome in adolescents/first year obtained with DNA degradation and lipid peroxidation assays. medical students of a public sector medical college in Quetta, Conclusions: In conclusion, aspirin, diclofenac sodium, and Pakistan celecoxib, with vitamin E supplementation, showed significant Faisal Shahzad1, Aiesha Ishaque2and Noman ul Haq3 synergistic effects that could be used as an effective preventive 1Bolan Medical College, Quetta, Pakistan,2Family Medicine, and therapeutic regimen for inflammatory diseases. Ziauddin University, Karachi, Pakistan,3Pharmacy Practice, University Clinical efficiency and cost effectiveness of macroprolactin of Balochistan, Quetta, Pakistan screening in hyperprolactinemic patients Introduction and objectives: Metabolic syndrome is a disorder N. Sherazi1, M.Z. Baig2, A.H. Khan1, I. Siddiqui1and F. Ghani1 of energy utilization and storage. According to the International 1Section of Clinical Chemistry, Department of Pathology and Diabetes Federation, metabolic syndrome in adolescents more Laboratory Medicine, 2Medical College, Aga Khan University Hos- than 16 years is defined as “the presence in an individual of central pital, Karachi, Pakistan. obesity plus any two of the following four risk factors: hyper- Introduction and Objectives: Macroprolactin (MaPRL) is a triglyceridemia, hypertension, low high density lipoprotein and biologically inactive compound which leads to falsely elevated high fasting glucose levels. prolactin levels. It is recommended that all sera with increased Reports suggest that around 25% of the world’s adult popula- total prolactin concentrations be sub fractionated by PEG pre- tion suffers from metabolic syndrome. People with metabolic cipitation to measure the bioactive monomeric prolactin con- syndrome are twice as likely to die and have three times higher centration to prevent misdiagnosis and unnecessary investiga- chances of heart attack or stroke. tions. We aim to determine clinical efficiency and cost effective- The study is designed to determine the prevalence of metabolic ness of PEG screening of hyperprolactinemic sera. syndrome in first year medical students between 17 and 19 years, Methods: In this Retrospective cross sectional study patients of a public sector medical college in Quetta. with high total prolactin levels were screened by PEG precipita- Methods: A Cross-sectional study was conducted on all the tion. Relevant diagnosis of Macroprolactinemia and True Hyper- healthy first year medical students, ages from 17 to 19 years prolactinemia was made based on their absolute PEG treated studying in a public sector medical college in Quetta, Pakistan. monomeric prolactin level. They were then contacted on phone Demographic data were assessed by using self-administered and a detailed history of their clinical symptoms along with their questionnaire. Height, weight and waist circumference were radiological workups was inquired. measured along with fasting blood sugar, high density lipoprotein Results: Frequency of macroprolactin was 60.7% seen in 145 and triglycerides by using Automated Biochemistry Analyzer. patients and true hyperprolactinemia was observed in 94 (39.3%) Analysis was done on SPSS 20. Descriptive was used to patients. More asymptomatic patients were reported in the mac- present the data and logistic regression was applied to see the roprolactin as compared to true hyperprolactinemic group relationship between the study variables (Po0.05). (pr0.05). 37 (39.4%) patients with true hyperprolactinemia had Results: A total of 225 were included in the study, out of which further radiological investigations done where-as only 8 (5.5%) of (n¼127, 56.4%) were females. Mean age was 18.79þ0.38 years. the patients with macroprolactin had undergone further radi- Prevalence of metabolic syndrome was found to be 14.2% in our ological workups. The total use of resources in the true hyper- study sample. Increased waist circumference, fasting blood sugar, prolactinemic group was significantly higher, 943000 PKR (10031 triglycerides were associated with an increased likelihood of ex- PKR per person) vs 418450 PKR (2884 PKR per person) (pr0.05). hibiting metabolic syndrome. Male gender and high HDL were Retinal nerve fiber layer thickness in a subset of Karachi found to be protective against metabolic syndrome. (Pakistan) population Conclusion: Prevalence of metabolic syndrome is increasing in Sahrish Mukhtar1, Nuzhat Hassan1, Zafar Dawood1and Nosheen our adolescent population which demands screening from child- Zehra1 hood and early adolescence. Early screening, identification and 1Ziauddin University, Karachi, Pakistan lifestyle interventions will decrease the morbidity from chronic Introduction and objectives: To provide the normal range of illnesses like Coronary artery disease and Diabetes mellitus. retinal nerve fiber layer (RNFL) thickness in a subset of Karachi Association of vitamin D with outcome after intra cyto- population by Spectralis OCT and to evaluate the effects of age and plasmic sperm injection N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 185

Umme Hani Abdullah1, Salima Lalani1, Sara Arif2, Fatima Syed3 Conclusion: Tegafer showed overall improved response rates, and Rehana Rehman3 cost effectiveness and has lesser side effects in comparison with 1Aga Khan University, Karachi, Pakistan,2Dow University of Health Capecitabine. Sciences, Karachi, Pakistan,3Jinnah Sindh Medical University, Karachi, Radiological study of the mean age of fusion of medial end Pakistan of the clavicle as a parameter of age Introduction and objectives: To observe effects of vitamin D Qudsia Hassan1, M Umer Memon1, Inayat-ur-RehmanKhalil1, levels on pregnancy outcome after intra cytoplasmic sperm in- Tehreem Ashraf1, Muhammad Mustafa Arif1and Ali Shan Iqbal1 jection (ICSI). 1Forensic Medicine, Ziauddin University, Karachi, Pakistan Methods: It was a cross sectional study conducted in Australian Introduction and Objectives: Estimation of the age of a person Concept Infertility Medical Center from July 2011 to August 2014. is a problem that a forensic expert confronts in both living and Estimation of 25 hydroxy cholecalciferol (25-OHD) of consented dead subjects. 21 years of age is of immense medico-legal im- females (252) was done before treatment protocol for ICSI. Results portance, as it entitles a person to be employed as a gazetted of- of ß hCG performed 14 days after embryo transfer categorized ficer, getting driving license for all types of vehicles, participation groups; Pregnant with ß hCG more than 25 IU/ml and rest in- in various sports events and is a mandatory parameter for iden- cluded in non-pregnant group. Both groups were compared by tification in dead. This study gives us an authentic criterion for independent sample t-test and Pearson Chi Square test. Binary assessing 21 years age, by using closure of ossification centers at Logistic Regression Analysis was used to estimate odds ratio of medial clavicular epiphysis by applying radiological method for pregnancy outcome with its predictors. subjects living in Karachi. Results: The mean value of 25-OHD, number of oocytes, ferti- Methods: The main objective of this study is to determine the lized oocytes and endometrial thickness was significantly higher mean age of fusion of medial end of clavicle by radiological in pregnant women. A significant positive association of 25-OHD method, to get a perfect estimator of 21 years of age. with clinical pregnancy and thickness of endometrium was ob- Results: The mean age of fusion of medial end of the clavicle served. After adjustment with female age and BMI positive asso- was found to be 2171.43 years. Significant difference was ob- ciation of vitamin D with endometrial thickness was observed. served in male to female ratio 21.1471.41 versus 20.6571.94 (P Conclusion: Deficiency of (25-OH) in females hinders the ac- value o0.05). Similarly, statistically significant difference was complishment of optimal endometrial thickness required for im- observed between lower class to middle class and higher class of plantation of embryo after ICSI. The improvement in vitamin D all society. No difference was observed between the various ethnic status can thus improve success results in assisted reproductive groups. clinics. Conclusion: Socio-economic factors such as diet and nutrition Comparison clinical outcome and cost-effectiveness of Te- directly affects bone growth and hence bone age. The results of gafur and Capecitabine for the treatment of metastatic color- fusion of the medial end of the clavicle are not affected by ethni- ectal cancer city. More studies should be conducted in various parts of country Khola Noreen1, Imran Shaikh1and Nadia Khalid1 to make a natural standard in setting up uniform criteria for as- 1Bahria University Medical and Dental College, Karachi, Pakistan sessing age at or above 21 years. Introduction and objectives: The development of numerous Centratherum anthelminticum (kalizeri) minimize the risk oral chemotherapy agents has led to a new paradigm in manage- of chemically-induced hepatotoxicity in rats. ment of cancer. Treatment modalities with cost effective approach Rabbiya Usmani1, Sumera Rais2, Syed Shabib Zaidi1and Shamim A and better clinical outcomes can be proved to be an effective Qureshi1 strategy to reduce disease burden. 1Department of Biochemistry, University of Karachi, Pakistan,2 To evaluate the clinical and cost effectiveness of oral che- Department of Biochemistry, of Arts, motherapeutic agents Capecitabine and Tegafer as first-line and Technology, Karachi, Pakistan treatments for patients with metastatic colorectal cancer. Introduction and Objectives: Carbon tetrachloride (CCl4) is a Methods: The analysis involved 121 patients of metastatic co- cleaning agent used in industry. Its vapours are injurious for many lonic carcinoma admitted at Almehrab Tibi Amdad Hospice care, body tissues especially liver. The study was designed to evaluate Karachi between September 2013 to March 2015. The patients the hepatoprotective effect of ethanolic seeds extract (ESEt) of selected to enrolled in present study were aged Z18 years, both Centratherum anthelminticum (kali zeri) in CCl4-induced hepa- sex and histologically diagnosed cases of colorectal cancer (stage totoxic rats. 3 with primary surgical resection. While those with history of Methods: Rats were made hepatotoxic by intraperitoneal in- hypersensitivity to Tegafur, fluoropyrimidines, capecitabine, or any jection of CCl4 (3 ml/kg) on 3rd and 5th day of trial and divided other ingredients of this product, Inadequate hematopoietic into hepatotoxic control (distilled water 1 ml/kg), positive control function WBC 4000/mm3; ANC 2000/mm3; Platelet 100,000/ (silymarin 100 mg/kg), and test (ESEt 800 mg/kg) groups. Normal mm3, Inadequate organ function, CNS metastasis, life expectancy control rats were also run. After 24 h of last dose of CCl4, body less than 3 months and those who were not willing to participate weights (BW) of all rats were recorded, sacrificed them to collect were excluded. All subjects fulfilling the eligibility criteria were blood and serum to analyze liver-specific parameters. In addition, randomly assigned to two groups, one group received oral cape- livers were dissected out carefully to estimate lipid peroxidation citabine (1250 mg/m2 twice daily for 14 days followed by one (LPO). week gap) and other received Capsule Tegofer 500 mg daily for 28 Results: ESEt (600 mg) not only significantly decreased the days followed by one week gap. All the patients were followed and percent reduction in BW of test rats but also improved the levels of response was observed after 3 months, 6 months and at the end of alanine transferase (ALT), asparate transferase (AST), alkaline year. phosphates (ALP), total bilirubin, total protein and albumin. Beside Results: Patients receiving Tegafer showed significantly less these, percent inhibition of LPO was also increased. incidence of alopecia, diarrhea and stomatitis (Po0.05) while in- Conclusion: The ESEt of C.anthelminticum was found as a he- cidence of hand-foot syndrome and grade 3/4 hyperbilirubinemia patoprotective and antioxidant agent that could be used for the were found to be significantly more as compared to capecitabine preparation of medicine in future. (Po0.0001). Cost of treatment per patient and per cycle using oral Spectrum of preneoplastic and neoplastic lesions of intes- Tegafer was less than that using capecitabine. tine in a tertiary care hospital of Karachi, Pakistan 186 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

Asma Shabbir1, Talat Mirza1, Asif Qureshi1and Abdullah Khalid1 systemic infections such as epiglottitis, meningitis, bacteremia/ 1Dow Institute of Medical College, Dow University of Health Sci- sepsis, bronchitis and chronic obstructive pulmonary disease and ences, Karachi, Pakistan otitis in young children. H. parainfluenzae, in contrast, is a sa- Introduction: Gastrointestinal malignancies are the global prophyte that colonizes the upper respiratory tract but it hardly oncological problems. It is therefore imperative to investigate the causes respiratory tract infections and only occasionally infectious spectrum of this pathology in Pakistan. endocarditis. But many strains are now becoming resistant to beta Objective: To generate the spectrum of premalignant and ma- lactam antibiotics. We aim to test the sensitivity pattern of Hae- lignant lesions of small and large intestine received at Dow Di- mophilus influenzae and Haemophilus parainfluenzae. agnostic Research and Reference Laboratory. Methods: Total 45 sputum samples were analyzed collected Methods: The study was conducted during 2009–2014. All the from different hospitalized patients. Specimens were inoculated cases of preneoplastic and neoplastic lesions of intestine received on blood agar plates and incubated in 5-% CO2 at 37 °C for 24–72 h. during the period of 6 years were reviewed. The data was sub- Gram stain were perform for direct examination and differentia- jected to statistical analysis using SPSS version 22. tion was done on the basis of availability of hemin(X) and NAD Results: A total of 64 cases were diagnosed as premalignant (V) factor, oxidase and slide agglutination test. Antibiogram was lesions of intestine consisting of ulcerative colitis (19/29.7%), done by CLSI method. Statistical analysis was also performed using adenomatous polyp (17/26.6%), dysplasia (14/21.9%) and adenoma SPSS version 17. (14/21.9%). Results: H.parainfluenzae showed such sensitivity pattern About 478 cases were diagnosed as malignant lesions of in- AMP75%, AMC80%, CRO80%, SXT42%, C92%, CIP/OFX84% while H. testine as: influenzae showed 100% sensitivity towards AMP, AMC, CRO, SXT, C and 92% sensitivity against CIP. i. Adenocarcinoma grade I (65/13.6%), grade II (283/52.2%), grade Conclusions: Proper diagnosis and selective use of antibiotics III (95/19.9%) can minimize the resistivity of these organisms. ii. Squamous cell carcinoma grade I (2/0.4%), grade II (7/1.5%), Association between interleukin 6 gene polymorphism and grade III (3/0.8%) human papilloma virus infection in oral squamous cell carci- iii. Metastatic adenocarcinoma (19/4.0%) noma patients iv. Neuroendocrine (4/0.8%) Mehwish Zafar1, Naila Hadi1, Saeeda Baig1and Nosheen Zehra1 1Ziauddin University, Clifton Campus, Karachi, Pakistan Conclusion: Our study showed ulcerative colitis as the com- Introduction and objectives: Human papilloma virus (HPV) monest premalignant lesion and grade II adenocarcinoma the and IL 6 gene polymorphism are proven independent risk factors most common malignancy of intestine. for development of Oral Squamous Cell Carcinoma. The objectives Comparison of antibacterial activity of Punica granatum of this study were to find out frequency of IL6 gene polymorphism and Syzyium cumini leave extracts. and HPV and to evaluate any association between HPV and IL6 Huma Urooj1and Aliya Hayat1 gene polymorphism in OSCC patients and its effect on disease 1Department of Microbiology, Jinnah University for Women, Kar- prognosis. achi, Pakistan Methods: In this cross-sectional study 140 OSCC patients (104 Introduction and objectives: Plants are the richest source of males and 36 females), aged 18yrs and above were selected. De- phytochemicals, not only protecting the plants itself but also tailed questionnaire was filled followed by sample collection. After benefits the mankind. Punica granatum commonly known as po- DNA extraction PCR analysis for HPV and RFLP for IL6 gene poly- megranate and Syzyium cumini known as Jamum are well known morphism was carried out. for its phenolic compounds having antioxidant potential. They are Results: Mean age of the patients was 43.5711.84 years (range used in traditional medicine for the treatment of diabetes, diar- 31–40 years). Majority (n¼45; 32.1%) belonged to Urdu speaking rheal diseases, antiparasitic and antibacterial infections. ethnic group and were habitual eaters of Pan (n¼87; 62.1%) and Methods: The aim of the study was to identify the anti- Gutka (n¼82; 58.6%). The most common site of OSCC was buccal microbial potential of two leaf extracts of Punica granatum and mucosa (61.4%). Most of the patients presented with grade II (55%) Syzyium cumini. fresh leaves were washed and ground with distil and late stages (stage III & IV) (55.7%) of OSCC. Out of 140 samples, water. The extract was obtained which was filtered by Whatman 12 (8.6%) tested positive for HPV gene with the following pattern #1 filter for the removal of particals. The antimicrobial activity was of IL 6 gene polymorphism; GG (46.4%), GC (39.3%), CC (14.3%). A performed by agar well diffusion method against E.coli, Bacillus significant association was observed between stages III & IV of subtilis, Klebsiella oxytoca, Salmonella typhi, Micrococcus leutus, OSCC and IL6 genotypes GC (P¼0.001) and CC (P¼0.002). Also Proteus mirabilis, Staph aureus and Candida albicans. PBS was strong positive association (P¼0.003) was found between HPV taken as negative while streptomycin as positive control. and IL 6 (P¼0.007) CC homozygote genotype. Results: It was found that the extract of Punica granatum was Conclusions: This study finds an association between HPV and inhibitory to all microorganisms. The highest zone of inhibition IL6 gene polymorphism in late stage OSCC patients suggesting was observed against Proteus mirabilis and E.coli (25 mm). Leaf rapid and aggressive progress of oral carcinogenesis. extract of Syzyium cumini do not show any antimicrobial activity. Comparison of virulence factors of isolated Candida spp Conclusions: Pomegranate leaf extract can be effective against from lower respiratory tract infections in hospitalized patients GIT infections. Jamun, as reported earlier, is well known to control Naheed A Irfan1*, Sumaira Javed1, Justina Abel1and Syeda Gufrana diabetes cannot be used to control infections. Nadeem1 Identification, differentiation and sensitivity pattern of 1Jinnah University for Women, Karachi, Pakistan Haemophilus influenzae and Haemophilus parainfluenzae Introduction and objectives: Candida a diverse group of fun- Naheed A.Irfan1*, Maira Khan1, Tehreem Gul1, AmnaShafique1, gus has bacterial and fungal properties and 80% commensals of Syyada Ghufrana Nadeem1 normal human body i.e C. albicans and C. tropicalis are involved in 1Jinnah University for Women, Karachi, Pakistan various diseases like oral thrush, vulvovaginal candidiasis etc. Introduction and objectives: H.influenzae is a pleomorphic when they get opportunity and they now are considered as the Gram-negative coccobacillus that is isolated predominantly from third most pathogenic strain causing various diseases in human. the respiratory tract and this opportunistic species may cause The objective is to compare the virulence properties of different N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 187

Candida spp. credence to the use of Aloe vera for the formulation of certain Methods: 126 sputum samples were collected from May to compounds that would coin new, effective and more potent an- August 2015 from hospitalized patients. Samples were inoculated timicrobial drugs to combat pathogenic microorganisms. on SDA. Identification was processed by gram staining, germ tube BIoinformatics analysis of histone-lysine N-methyltransfer- test, Chlamydospore detection and CHROM agar. Phospholipase, ase - SETD1A gene in association with schizophrenia proteinase and hemolytic activity were delibrated in them. Sta- Zoobia Anis1, Hajira Karim1and Afshan Zeeshan Wasti1 tistical analysis was also performed using SPSS version 17. 1Department of Biochemistry, Jinnah University for Women, Kar- Results: 51% males and 48% female were found to be infected achi, Pakistan with Candida spp. The most prevelant organism was C. albicans Bioinformatics methodologies have made it possible to profile (87.5%) followed by C. glabrata (8.56%) and C. tropicalis (3.08%). the global composition of tissue or organelle at specific time point Phospholipase activity (45.9%), proteinase (30.1%) and haemolysin or under particular developmental or disease state. A genetic activity (19.9%) was shown by C.albicans. Phospholipase and pro- variant of SETD1A has yielded significant genome-wide associa- teinase activity was also observed in C.tropicalis but not in C. tion with schizophrenia, suggesting that this SETD1A plays a key glabrata. role in its etiology. The SETD1A gene plays a part in chromatin Conclusions: Present study indicate that C.albicans being a modification - an important cellular process that reduces the size member of normal flora pose a major threat in immune-compro- of DNA so it can fit inside a cell and regulate gene expression. It mised patients and majors should be inforced to prevent infections encodes a catalytic subunit of an enzyme that adds methyl groups with Candida. to histone proteins - genes for transcription. Structural investigation of Neisseria meningitidis patho- The present study is based on computational analysis of SET- genic factors; a key step towards drug designing D1A gene exploring molecular networks of interacting targets by Sana Aurangzeb1, Mehwish Hamid1and Yasmeen Rashid1 using advanced bioinformatics tools such as, STRING’s, KEGG, Re- 1Department of Biochemistry, University of Karachi, Karachi, actome and BioGrid databases that may provide insights into the Pakistan biological processes underlying schizophrenia. Introduction and objectives: The Gram negative human pa- Frequency of Hepatitis B and C in different area of catch- thogenic bacterium Neisseria meningitidis is responsible for ment center in , Karachi, Pakistan. causing meningitidis and septicemia worldwide. A substantive Aliya Zaman1, Nazish Jaffer2and Rubina Ghani3 question in bioinformatics analysis of bacterial genome is to as- 1,2Department of Pathology, Sir Syed Medical College, Karachi, cribe a three dimensional structure as well as a biologic role to all Pakistan2Jinnah Sindh Medical University, Karachi, Pakistan,3De- the coding regions. Here we performed structural bioinformatics partment of Biochemistry, Jinnah Medical and Dental College and analyses of important Neisseria meningitidis pathogenic factors Pathological and Molecular Laboratories, Karachi, Pakistan involved in protein biosynthesis including methionyl-tR-NA syn- Introduction and objectives: Frequency of Hepatitis B Surface thetase, 16SRNA methyltransferase, translation elongation factor- Antigen (HBs Ag) and Hepatitis C Virus Antibodies (anti-HCV) Tu and putative RNA methylase. among population of different catchment center in Gadap area Methods: Homology modeling of these important drug targets with their low social economic status. We aim to study the fre- was carried out with better templates by MODELLER software and quency of Hepatitis B Surface Antigen (HBs Ag) and Hepatitis C evaluated by Prosa and Procheck standalone softwares. Virus Antibodies (Anti-HCV) in different catchment center of Ga- Results: The study provided detailed structures of important dap town in 12 kilometer from Baqai Medical University. The de- proteins required for the pathogenesis of this organism. sign of study will be cross sectional descriptive study. It was held Conclusions: In future, these investigations will help in re- in the Pathological and Molecular Laboratories, Karachi, during the ceptor-based drug designing against meninigococcal infections. period October 2011 to May 2012. Evaluation of antimicrobial activity of Aloe vera. Methods: A total of 496 samples were collected. Serum was Iqra Waqar1and Aliya Hayat1 tested for Hepatitis B surface antigen and for Hepatitis C virus 1Department of Microbiology, Jinnah University for Women, Kar- antibodies by ELISA. The results were subjected to chisquare achi, Pakistan analysis for determination of statistical difference between the Introduction and objectives: Aloe vera is a cactus, belongs to values among different categories. the family Liliaceae, and is well known for its use as traditional Results: Among 496 patients 91 (18.3%) were positive for HB s medicine. It is use for gastrointestinal disorders, stimulates body’s Ag and 171 (34.4%) were positive for Anti-HCV. The frequency of immune response, for treatment of burns, eczema, psoriasis, to HB s Ag was seen too low as compared to anti-HCV. heal skin infections. Aloe vera is known to hold therapeutic Conclusions: High frequency of HCV infection needs im- properties due to bioactive components present in it for both in- plementation of strict screening policy for donors and public fectious and non-infectious diseases. awareness campaigns about preventive measures to reduce the Methods: Antimicrobial activity of Aloe vera was investigated spread of this infection as well as other transfusion transmissible against Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, infections. Micrococcus, Klebsiella pneumonia, Proteus, Staphylococcus aur- Innovative approaches in metabolomics for understanding eus, Salmonella typhi and Candida albicans. The organisms were drug resistance in breast cancer isolated from clinical samples and identified by conventional Umber Saeed1and Usman Choudhary2 methods. Susceptibility test was carried out by agar well diffusion 1Dow College of Pharmacy, DUHS, Karachi, Pakistan,2London method. Aloe vera used as methanolic extract and pure gel. All PGDE, Scotland, UK experiments were performed in triplicate. Introduction and objectives: Breast cancer is one of the Results: Zone of inhibition was observed in all forms of Aloe leading cause of death worldwide. In Pakistan, prevalence of this vera. The maximum zone of inhibition was found in E.coli and ailment is highest amongst all types of cancer i.e. 38.5%. Major followed by Pseudomonas aeruginosa, Salmonella typhi, S aureus clinical setback is drug resistance in breast cancer. Metabolomics is and Bacillus subtilis showed least zone of inhibition. It was also an emerging field that utilizes information of cellular biochemistry found that inhibitory affect against Candida albicans was also for the early detection, diagnosis and establishment of predictive maximum. biomarkers of breast cancer. This review highlights potential me- Conclusions: The result of this study would tend to give tabolomic applications to pharmacology and clinical 188 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 pharmacology. detailed structural analyses. These proteins are also involved in Methods: The methodology is based on inclusion exclusion RNA-degradosome assembly. criteria. Literature survey, and questionnaire were included while Results: Different bioinformatics strategies were applied and clinical trial was excluded. This report provides a review of 12 the homology models of these proteins were built using protein articles out of few were excluded. structure-modeling program MODELLER and the models were Results: According to the survey the average response rate of a evaluated using PROSA and PROCHECK software, as well as active cancer drug is the lowest at 21%, suggesting that 74% of patients sites are also predicted. with cancer are over-dosed. While according to an international Conclusions: Since these proteins are potent dug targets, study, 40–50% of breast tumors will display acquired resistance. therefore their structural identification will prove to be a break- Conclusions: When specific therapies are chosen on the basis through in drug discovery. We hope that our study will grease the of a patient's metabolomics profile, it will give rise to customized wheels for drug designing against lethal meningococcal disease. medicine and personalized tailored treatment. Using high- Role of rauwolfia serpentina extract on the modulation of im- throughput information using metabolomics to clinical diagnosis mobilization stress-induced behavioral deficits and adaptation to and treatment can help accelerate the patient safety, quality of life stress in rats and survival rate by identifying pathways involved in drug Erum Shireen1, Wafa Bint E. Ali1and Darakshan J.Haleem1,2 resistance. 1Department of Biochemistry, Neurochemistry and Biochemical Aberrant protein S-nitrosylation in hypertensive and dia- Neuropharmacology Research Laboratory, University of Karachi, betic-hypertensive patients: New perspective Pakistan,2Neuroscience Research Laboratory, Dr Panjwani Center for Erum Zafar1, Ayesha Khan1, Beena Hasan1and Nikhat Ahmed1,2 Molecular Medicine and Drug Research, University of Karachi, 1Neurochemistry Research Laboratory, Department of Biochem- Pakistan istry, University of Karachi, Pakistan,2Research Department, Ziauddin Introduction: Stress is the major precipitating factor in the University, Karachi, Pakistan onset of depression, anorexia nervosa, diabetes and obesity. The Hypertension and diabetes are the risk factor of morbidity and goal of present study is to monitor the behavioral and biochemical mortality. The prevalence of hypertension is expected to increase effects of post stress administration of Rauwolfia serpentina ex- in the upcoming years. The incidence of diabetes is two fold higher tract on adaptation to immobilization stress in rats. in patient with hypertension. The co-existence of diabetes and Methods: The plant extract (30 mg/kg) was orally administered hypertension causes impaired renal function, the development of after termination of immobilization stress daily (2 h) for 5 days to diabetic retinopathy, cardiac problems, and the development of monitor any change in behavioral activities. Effects of Rauwolfia cerebral diseases. Besides these disorders hypertension and dia- serpentina extract on immobilization stress induced deficits of betes are involved in many other devastating disorders. Protein food intake and body weight were determined for an under- S-nitrosylation conveys a large part of the ubiquitous effect of standing of the relationship between stress tolerance and beha- nitric oxide on cellular signal transduction, accumulating evidence vioral changes. We also tested effects of Rauwolfia serpentina indicates important roles for S-nitrosylation both in normal phy- extract on endogenous leptin and glucose levels in unstressed and siology and in a broad spectrum of human diseases. Dysregulated stressed animals to explore the possible role of HPA axis in the S-nitrosylation has been implicated as a cause or consequence of a modulation of stress-induced behavioral deficits and adaptation to broad range of diseases, including asthma, cystic fibrosis, Parkin- stress. son disease, heart failure, and stroke. The role of nitrosylases and Results: The present study showed that Rauwolfia serpentina denitrosylases in governing levels of S-nitrosylation under both extract can blunt stress-induced anorexigenic as well as anxio- physiological and pathophysiological conditions is increasingly genic-like effects. Moreover, Rauwolfia serpentina extract reversed appreciated. The purpose of study is to identify s-nitrosylation in adverse effects of stress and facilitate adaptation to an un- hypertensive and diabetic hypertensive serum samples. We take controllable stressor by reducing stress perception. serum sample of hypertensive subjects (n¼10), diabetic hy- Conclusion: The present study shows that immobilization pertensive subjects (n¼10) and normotensive subjects (n¼10). stress induced decreases in food intake, body weight as well as Total protein was quantified using Bradford assay. SDS PAGE was behavioral deficits were reversed by Rauwolfia serpentina extract done after dilution of sample followed by western blotting. The suggesting anxiolytic like profile of drug. This effect of Rauwolfia results we obtained from the above mentioned methods showed serpentina extract can be explained in terms of additive effects of that s-nitrosylation is decreased in hypertension and slightly in- stress on serotonin (5-hydroxytryptamine; 5-HT) neurotransmis- creased in diabetic hypertension. sion particularly via postsynaptic 5HT2C as well as 5HT1A re- Structural bioinformatics analysis of transcription factors ceptors. It is therefore suggested that post stress administration of involved in Neisseria meningitidis pathogenesis; a leap towards Rauwolfia serpentina extract provides an innovative approach for antimeningococcal drug discovery the treatment of stress related disorders. Mehwish Hamid1, Sana Aurangzeb1and Yasmeen Rashid1 Alteration in salivary parameters lead to oral lesions among 1Department of Biochemistry, University of Karachi, Karachi, chewable tobacco users Pakistan Areeba Asghar1, Saeeda Baig2, Asghar Ali1and Muhammad Asif Introduction and objectives: Neisseria meningtidis is a viru- Memon1 lent pathogen causing meningtitis as well as life-threatening 1Department of Physiology, Ziauddin University, Karachi, septicemia throughout the world. Unfortunately, any vaccine or Pakistan,2Department of Biochemistry, Ziauddin University, Karachi, drug has not yet been developed against N.meningtidis serogroup Pakistan B. Functional genomics strategies have been adapted to study the Introduction: Salivary flow and composition alters under de- growth and pathogenesis of Neisseria meningtidis in which a li- leterious chemical irritants present in chewable tobacco. Apart brary of 2850 insertional mutants was analyzed and 73 genes were from inflammation and deterioration in health of oral mucosa it identified in N.meningtidis genome responsible for causing can also affect the major and minor salivary glands and cause a disease. decline in salivary flow rate. The objective of the study was to find Methods: We have selected the proteins (gene products) i.e., out alteration in salivary parameters that lead to oral lesions Dead box RNA-Helicase, Polyribonucleotide nucleotidyl-transfer- among chewable tobacco users. ase PNPase and Ribonuclease-III involved in transcription for their Methods: A total of 354 healthy male subjects, consuming any N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 189 form of chewable tobacco product, belonging to low socio- 1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, economic areas of Karachi were selected for this cross sectional Ziauddin University, Karachi, Pakistan,2Department of Pharmaceu- study. A questionnaire was used to collect demographic data and tical Chemistry, Faculty of Pharmacy, University of Karachi, Karachi, details of chewing habits (using since, pack/day, duration of ex- Pakistan,3Baqai Medical University, Karachi, Pakistan posure etc.). Resting saliva of every subject was collected for 5 min A pharmaceutical formulation ensures safety, protection, pro- and RSFR was expressed in ml/min. Salivary pH was determined curement and preservation of human health. Therefore, pre- by using pH strips (pH 0–14). Oral examination was done for the servation of its physical and chemical stability throughout the presence or absence of oral lesions. Data was analyzed on SPSS period of its shelf life has always been the matter of prime concern version 20. for a pharmaceutical manufacturer. A good formulation is one that Results: Out of the 354 subjects included, 27.4% consumed when packaged in a suitable container, (which provides maximum gutka, 24.3% niswar, 24.3% paan and 24% were multiple users. protection to its contents), confirms the absolute delivery of its Mean Resting Salivary Flow Rate (RSFR) was 0.5270.34 ml/min, active ingredient in its original form to its consumer. To determine pH 6.5870.78% and 27.1% (n¼96) had oral lesions. Highest fre- the suitability of blister strips for packaging of Metformin HCl, quency of oral lesions was found among subjects who had hypo- Amlodipine besylate and Ciprofloxacin hydrochloride tablets, we salivation (40%) and those having acidic pH (40%). A significant maintained a stability profile for the changes in physical appear- decrease in RSFR and pH and increase in frequency of oral lesions ance and loss of potency of these for a period of 2 shelf is observed with increased duration of exposure, duration of usage life years. Tablets were subjected to the specified conditions of and increased number of tobacco packs consumed per day. 30 °C and 65% RH for 24 months and samples were withdrawn at Conclusion: Increased frequency and recency of chewable to- different time intervals. Different tests and assays were conducted bacco use leads to decrease in RSFR and pH and hence increase in on these samples showed that under these specified conditions frequency of oral lesions. the efficacy of these tablets in the blister strips retain for less than Speech assessment through thetri-position articulation 2 shelf life years which is less than the expected shelf life of analysis for bilinguals (TAAB) 3 years. Amina Asif Siddiqui1 Retrospective analysis of maternal mortality at Ziauddin 1College of Speech Language and Hearing Sciences, Ziauddin UNIVERSITY Hospitals, from 2012–2014 University, Karachi, Pakistan Chaudhry Shama1 Introduction: Bilingualism is regarded as a cognitive feat by 1Ziauddin University, Karachi, Pakistan today's linguists. The acquisition of more than a single language Introduction and objective: To determine the frequency of manifests the miracle of the human brain in terms of meta lin- maternal mortality and identify the causes with different age guistic (meta phonological) skills, as it stores the form and content groups, parity and gestational ages in Ziauddin university hospitals of multiple languages and exhibits their use in socially and lin- in 4 campuses, from 2012 to 2014. guistically appropriate contexts. Methods: A descriptive study conducted at Gynecology & Ob- Methods: Pakistan, like most Asian countries has a pre- stetrics units of Ziauddin university hospitals with retrospective dominantly bilingual fabric. Little children, regardless of their so- review of validated records of hospital registers in 3 years from cioeconomic status are most often exposed to their native lan- January 2012 to December 2014. Convenience sampling used to guage (L1) and to a second language (L2) within the critical period access the records. A self-structured proforma designed to collect (0-3years), leading to simultaneous bilingualism. When they are data on variables (age, parity, gestational age, cause of death, exposed to L2 after the acquisition of L1 we call it sequential bi- condition at the time of admission, duration of stay in hospital lingualism. Some children may have L3 in their linguistic re- before death, cause of delay). pertoire which may be the additional language such as the lan- Results: During 3 year period from January 2012 to December guage of instruction at school. The development of articulation 2014 there were 32 maternal deaths. Total number of births were and phonological skills in children usually reaches maturation 14219 and 14184 were live births. Still births were 33. Eclampsia around the chronological age of 6 years. Children pick up several (21.8%) and hemorrhage (21.8%) were the leading cause of ma- sounds of their language and perfect them gradually using ap- ternal mortality followed by those with puerperal sepsis (12.5%) proximations that sound closer and closer to the target sound, and others. MMR (maternal mortality ratio) was 226 per 100000 acquiring all sounds with the culmination of early childhood. live births. Results: Children may present with unclear speech, in the ab- Conclusion: Maternal mortality was high with leading causes sence of an organic etiology (dyslalia or phonological disorders) or as hemorrhage, eclampsia, and sepsis in ages of 20–30 years and because of a known organic cause such as cleft lip/palate, mal- parity 1–4. Mostly deaths were due to delay in getting the women occlusions, hearing loss, dyspraxia or dysarthria, must be assessed to the health facility. for their ability to acquire phonemes of all the languages in their Vitamin D deficiency in pregnant women and their new repertoire, because the brain stores auditory images of all sounds born and uses them appropriately when needed. Shazia Sultana1 Conclusions: The Tri-position Articulation Analysis for Bilin- 1Ziauddin University and Hospital, Karachi, Pakistan guals is a tool for testing the articulation and phonological skills of Introduction and objectives: Vitamin D deficiency may affect bilingual children aged 0–6 years, that uses culturally and lin- the mother and its new born health. The aim is to detect the fre- guistically appropriate picture stimuli for children exposed to quency of vitamin D deficiency in pregnant women and their Urdu, Sindhi, Punjabi, Pushtu, Hindi and English. The test assesses newborn of Karachi at tertiary care hospitals, and correlating spontaneous elicitation of the sounds in the child's lexicon, along maternal vitamin D with cord blood. with stimulability and also assesses the function of the speech Methods: This cross-sectional study was conducted on 50 mechanism for non-speech functions. women in labor presenting with a singleton term pregnancy at Stability studies of metformin hydrochloride, Amlodipine tertiary centers in Karachi. Data was recorded on a special pro- besylate and Ciprofloxacin hydrochloride tablets under speci- forma, maternal blood was taken before delivery and cord blood fied condition of temperature and relative humidity of was taken at delivery. All blood samples were analyzed for 25- Pakistan hydroxy vitamin D levels. Qurratul ain Leghari1, Sohail Hassan2and Iqbal Ahmed3 Results: The mean vitamin D levels were 24 ng/ml for the 190 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 mothers and 20 ng/ml for the newborns. Vitamin D sufficiency Clifton laboratory Karachi, Pakistan over a period of 7 months from was noted in 11 (22%), insufficiency in 16 (32%), and deficiency in 1st June until 31st Dec 2014. Healthy blood donors and patients 23 (46%) of the 50 mothers whereas sufficiency and deficiency, requiring blood products of different age groups were employed. were noted in 6 (12%) and 44 (88%) of the newborns respectively. After an informed consent blood grouping was performed by There was a positive correlation between the vitamin D levels in taking blood samples under aseptic measures in EDTA and Gel maternal and cord blood (r¼0.03; P¼0.003). Maternal vitamin D separating serum tubes for ABO & Rh blood grouping by tube levels were significantly affected by sunlight exposure (P¼0.007) method. and quality of diet (P¼0.01). Results: Total 1583 individuals included 868 (54.8%) males and Conclusion: Vitamin D deficiency is high among pregnant ur- 715 (45.2%) females. Oþve blood group found to be more common ban Pakistani women and their newborns. This public health group (31.9%) followed by Bþve (31.2%). In Rh system, Rhþve problem needs urgent attention. (97.2%) were more common than Rh-negative (2.8%) blood groups. Are you prepared to care your loved ones: A cross sectional Conclusion: This study was taken out to maintain the blood study to assess the level of preparedness among caregivers of product inventory concerning the management of blood bank and chronically ill patients in a tertiary care hospital at Karachi, transfusion services for the patient. Pakistan Frequancy of ABO blood group discrepancies in tertiary care Shireen Arif1 hospital Karachi and their classification into groups 1College of Nursing, Ziauddin University, Karachi, Pakistan Sumera Shaikh1, Saba Jamal1and Fatima Mairaj1 Introduction: Family caregivers are also known as informal 1Ziauddin Hospital, Karachi, Pakistan caregiver. They provide all level of care at home to their loved ones Introduction: In developing countries, the blood grouping is most often equivalent to the professional care providers, such as generally being performed by the forward grouping method only. providing physical care, giving recognizing early signs The aim of this study was to find out the frequency of ABO blood of severe conditions, changing dressings etc. While providing care group discrepancies in tertiary care hospital in metropolitan city of to their loved ones most of them feel unprepared in terms of lack Pakistan and also to classify these discrepancies on the etiological of knowledge and skills. This unpreparedness may result in the basis. role strain and have may psychological consequences for the fa- Methods: Cross-sectional, descriptive type of study was con- mily. Nurses as a primary care provider could assess the caregiver ducted at Blood Bank of Dr. Ziauddin Hospital Karachi, Pakistan. preparedness and may educate them according to the patient’s Total 1522 samples were included with exclusion of newborns and need to reduce the extra burden and the role strain. infants up to the age of 6 months. Blood samples were collected Method: The self-administered instrument regarding the pre- from a peripheral vein by trained phlebotomist and submitted to paredness for care giving scale by Archbold (1990) has been uti- the blood bank department in 2 tubes, purple top vacutainer tube lized. The scale has eight items to identify the preparedness of containing Di-potassium Ethylene Diamine Tetra Acetic Acid (K2 caregiver in multiple domains. The responses are rated on five EDTA) anticoagulant and red top vacutainer tube containing clot- point scale with scores ranges from 0 (not at all prepared) to 4 ted blood. Blood group testing was performed by tube technique. (very well prepared). The higher score means caregiver feels more Results: In total 1557 patients (854 Males and 703 Females) preparedness and vice versa. 50 caregivers of chronically ill pa- from various clinical specialties were included for ABO blood tients were selected at a tertiary care hospital, which were caring grouping. The male to female ratio was 1.2:1 (854 males and 703 for their loved ones for about six months or above. Written con- females) with age ranging from 9 months to 92 Years. ABO dis- sent was obtained from the participants. crepancies were Positive in 18 out of 1557 Patients (1.1%). Male to Result: 50 caregivers were recruited to fill the form from which female ratio among ABO discrepancies were 1.5:1. There were four 26 were female and 24 were male. All of the participants were groups of ABO discrepancies. The most common group of dis- between 18 years and 60 years of age. About 68% caregivers score crepancies were group I discrepancies consisting of 12 out of 18 between not too well prepared and somewhat well prepared. Patients (66.7%). Group II discrepancies were consist of 2 out of 18 Whereas, only 32% score between somewhat prepare and pretty Patients (11.1%). Group III discrepancies were consisting of 1 out of well prepared. 18 Patients (5.5%) and Group IV discrepancies were consisting of Conclusion: In conclusion, most of the caregivers score are 3 out of 18 Patients (16.7%). somewhat prepared which is an alarm for nurses and all health Conclusion: The forward and reverse grouping are the essen- care providers to work to reduce their role strain and provide tial parts of blood group testing for assigning the correct ABO sessions according to the need of the caregivers. Nurses could ar- blood group to the individual. ABO discrepancies i.e. forward and range teaching sessions which could be helpful for the caregivers reverse grouping are not matches to each other can cause severe to do their role effectively. transfusion reactions which can be fatal for life. Furthermore, to ABO and Rhesus blood group distribution in residents of ensure that correct ABO blood product to be transfused for saving Karachi life. Therefore reverse grouping should be strictly adopted in every Sumera Shaikh1, Shariq Adnan1, Mustafa Zuberi1and Syed Sohail blood bank and transfusion services as a routine practice. Abbas1 Selfmedication practice among undergraduate medical 1Ziauddin University, Karachi, Pakistan students Introduction and objective: Blood is an essential component Tahira Asad1, Tahira Zamir1, Nasim Karim1and Talea Hoor1 for human beings and in disease states, it is requested to be 1Ziauddin University, Department of Pharmacology and Ther- transfused from healthy human donors. But it was continuously a apeutics, Medical and Dental College, Karachi, life threatening procedure until the discovery of ABO blood Pakistan groups. Today committee of League of Nations has recommended Introduction and objective: Assessment of self-medication A, B, AB & O classification for general use. This study was aimed to practice among medical students is presumed to be of exceptional determine the frequency of ABO and Rh blood groups. So that importance as in future they have promising role in counseling the necessary measures should be taken to maintain the blood pro- patients regarding the rational use of drugs. This study was spe- duct inventory. Frequency of ABO and Rhesus blood group dis- cially designed to evaluate self-medication practice in 3rd year tribution in residents of Karachi. medical students. Methods: A study was conducted at Dr. Ziauddin Hospital Methods: A cross sectional institution based study was N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 191 conducted from Jan – March 2013 on 3rd year medical students at samples for estimation of Kisspeptin, was done on OI day and Bahria University Medical and Dental College, Karachi, after ap- endometrial thickness was measured. Based on beta hCG results proval by ERB-BUMDC. Data was collected by questionnaire, filled were categorized into group A, non-pregnant with beta hCG on one to one basis after taking verbal consent and analyzed on o25 m I U/ml II, and group B, clinical pregnancy with beta SPSS version 16. hCG45 m IU/ml. Results: Frequency of self-medication was found to be 90%. Results: Kisspeptin levels were significantly higher in the Quick relief (38%) and busy schedule (21%) were the main reasons Group B versus Group A (po0.001) independently associated with provoking self-medication. Common ailments that led to self- positive pregnancy (r¼0.388; po0.001), and endometrial thick- medication were: headache (95%) and fever (87%). Drug groups ness (r ¼0.294; p¼0.05) irrespective of the age and BMI of the frequently used were: analgesics (95%), antipyretics (87%) and subjects. antibiotics (36%). Acetaminophen was the most common analgesic Conclusion: Kisspeptin is a positive pregnancy marker in fe- (73%) as well as the antipyretic (88%) while antibiotics were, me- males after ICSI as a result of its effects on oocyte maturation and tro-nidazole (26%) followed by amoxicillin plus clavulanic acid endometrial thickness. (17.3%). Effect of long hour shifts on medical interns and residents, Conclusion: Practice of self-medication is high among medical and the strategies adopted to cope with work-related stress students. Potential problems of self-medication should be em- Maryam Kazim Dattoo1, Nimra Shah1and Aliza Fatima1 phasized to the students. 1Ziauddin University, Karachi, Pakistan Determination of prostate gland volume by ultrasonography Introduction and objectives: To assess the impact of long hour and its correlation with anthropometric measurements in a shifts on performance and health of medical interns and residents subset of Karachi population and strategies adopted to cope with stress. Iffat Raza1 Method: This cross-sectional study was conducted from Jan- 1Department of Anatomy, Ziauddin University, Karachi, Pakistan uary to July 2015. A sample of 200 interns and residents was taken Introduction and Objectives: To establish the local reference from Ziauddin Hospital Clifton, Kemari and North Nazimabad. Data range of prostate volume according to our subset of population. To was collected through a self-designed questionnaire and stress correlate prostate volume (PV) with age, body mass index (BMI) levels assessed through General Health questionnaire (GHQ-12). and waist circumference (WC). Data was analyzed through SPSS v 20. Chi-square and Paired T test Methods: A cross-sectional study with 119 healthy adults aged was applied. 40–79 years without any prostatic pathology were recruited. The Results: The mean age of participants was 27þ2.3 years. study population was categorized into 4 age groups(40 - o50 Average sleep hours when doing call were 3.06þ1.47. Work per- yrs,50 -o60 yrs,60-o70yrs,70-o80 yrs), 3 BMI groups (healthy, formance was below average in 62% of the population. Stress was obese and overweight) and 2 WC groups (o90 cm and 490 cm). found in 34% of study population. Increased tea intake was the A p-value of o0.05 was considered significant. A total of 130 stress coping strategy adopted by 72%. There was a significant healthy adult volunteers were recruited initially, out of these 20 association between stress level, gender and personal habits. individuals were those whose IPSS was o8 and prostate volume (pr0.05). A significant difference was found between cigarette on ultrasound was 425 ml. These 20 subjects underwent Uro- and energy drink intake before and after starting job (pr0.05). flowmetry (UFM). After UFM, total 19 subjects were labeled as Conclusion: High stress may have negative effects on health healthy adults and 1 subject was excluded from healthy adult and performance. Hence, medical interns and residents need group. The sample of 119 healthy subjects were studied sono- support to cope with stress. graphically for the effects of Age, BMI and WC on prostate volume. Recurrent miscarriage and associated factors Results: The mean prostate volume was 21.772.2 mls, mean Rehana Yasmeen1and Rubina Hussain1 body mass index was 2876 kg/m2, whereas mean waist cir- 1Ziauddin University, Karachi, Pakistan cumference was 95 cm. PV was found to be higher in obese and The incidence of recurrent pregnancy loss in women of re- 490 cm waist circumference group. After applying multiple re- productive age group is 0.5–3%, and idiopathic causes accounts for gression analysis, waist circumference correlated positively and 50–60% of recurrent pregnancy losses. Approximately 30–50% of significantly with prostate volume. UFM showed negative corre- conceptions end in spontaneous miscarriage before the comple- lation with IPSS and PV. tion of first trimester. Miscarriages mostly occur at the time of Conclusions: Mean prostate volume in our studied population implantation. The causes of recurrent pregnancy loss are many was smaller than that of many western populations. Our study has including parental chromosomal abnormalities, maternal throm- proved that central obesity is the most important factor influen- bophilias, immunologic causes and endocrine disorders. Recurrent cing prostate volume. pregnancy loss is an extremely distressing clinical problem for Association of Kisspeptin, endometrial receptivity and un- women as well as health professinals. According to current studies explained infertility the decidualized endometrium act as a biosensor of good quality Rehana Rehman1, Zehra Jamil1, Syeda S Fatima1, Sara Arif2and embryos, if it is faulty, may lead to implantation of embryos re- Faiza Alam1 sulting in miscarriage. Other factors implicated in the pathophy- 1Department of Biological and Biomedical Sciences, Aga Khan siology of miscarriage are Systemic and placental oxidative stress. University, Karachi, Pakistan,2Civil Hospital, Karachi, Pakistan Vascular endothelial damage, abnormal vascularity of placenta and Introduction and objectives: To relate serum Kisspeptin levels immunologic reactions have been proposed to play some role in with pregnancy outcome after intra cytoplasmic sperm injection causing recurrent miscarriage. (ICSI). The objective of this review is to discuss the causes of recurrent Methods: In the cross sectional survey carried at Australian miscarriage, based on the published research articles. Concept Infertility Medical Center from June 2014 to June 2015, Prevalence and preventive measures of prostate cancer: A 176 females with 20–42 years of age and with regular menstrual randomized study in public and private health care sector cycles were included for ICSI. Patients with uterine fibroids and Somia Gul1, Umm-e-Aimon1and Maria Ayub1 metabolic disorders were excluded. Down regulation of ovaries 1Faculty of Pharmacy, Jinnah University for Women, Karachi, was followed by controlled ovarian stimulation, ovulation induc- Pakistan tion, oocyte retrieval, microinjection and embryo transfer. Serum Introduction and objectives: Prostate cancer accounts for 192 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 abnormal or uncontrolled division of cells of prostate in male. It is Dimensional Scale of Perceived Social Support, Internalized Stigma a slow growing cancer which is usually localized rarely metasta- of Mental Illness Scale (ISMI) and Rosenberg Self Esteem Scale sized. The growth rate varies from slow moderate to high. It were administered individually. mostly occurs in old age and depends on many other factors like Results: Findings revealed social support predicts internalized weight, height, diet, habit, environment, , and sexual dis- stigma and self-esteem in persons with schizophrenia (po0.000). ease. This study aims to evaluate occurrence, causes, problems Conclusions: There is an immense need to develop awareness associated with disease and treatment of this cancer in view of the regarding social support group and to generate strategies to re- fact to evaluate consequences responsible to contribute carcinoma duce the effect of internalized stigma in person with severe mental of prostate. illness. Methods: For this purpose randomized study conducted at Efficacy and safety of Metformin in gestational diabetes public and private health care sector (n¼100 patients of prostate mellitus cancer). Huma Muzaffar1, Shama Chaudhary1and Rubina Hussain1 Results: The Gleason score are used to diagnose prostate can- 1Ziauddin University, Karachi, Pakistan cer, 10% of patients have score 5 (4þ1), 40% of patients have score Objective: To assess the efficacy and safety of Metformin for 7(4þ3) or (3þ4), 44% of patients have score 9 (5þ4), 6% of pa- the management of gestational diabetes Mellitus (24 wks and tients have score 11 (5þ6) which shows that mostly prognostic above) as compared to Diet. grade II, III IV, V occur. Mostly traditional prostectomy is used as a Methods: This Quasi experimental study is being conducted on treatment and 78% of the patients got benefit from it. 600 females at Ziauddin University Hospital. All pregnant women Conclusions: This study concludes that the prostate cancer receiving prenatal care fulfilling the inclusive criteria will be may occur due to age factor (old age), fatty diet, tobacco or alcohol screened using a 50 g glucose challenge test (GCT). After the result intake. Finding of cancer at score 5,7,9,11 and associated compli- if it is positive, women will be advised for oral glucose tolerance cations are hypertension, urinary tract infection, kidney stones. test. After diagnosis of gestational diabetes Mellitus patients will Knowledge and attitude towards menopause in women aged be advised for diet control or oral hypoglycemic drug metformin 45–60 years starting from 500 mg twice a day. Patient will be followed fort- Rehana Yasmeen1, Rubina Hussain1, Shama Chaudhry1and Aliya nightly with sugar record for adjustment of treatment. Patient will Waheed1 be followed till delivery for any maternal or perinatal complica- 1Ziauddin Hospital North Nazimabad, Karachi, Pakistan tions. Patients with systemic diseases are excluded from the study. Introduction and objectives: Menopause has significant phy- Results: Result of this study are awaited as this is an ongoing sical, mental, social and psychological effects on women’s life and study. prior knowledge about these changes help them to cope with Conclusions: The main purpose of the treatment is to prevent these changes with greater readiness. Educating women regarding fetal hyperinsulinemia and fetal macrosomia by reducing maternal menopausal changes would definitely equip them better to toler- glucose level, decrease risk of appearance of diabetes in sub- ate these menopausal symptoms. sequent pregnancies and in future without pregnancy. Metformin Methods: A cross sectional study based on sample of con- is an alternative to insulin and is effective in the treatment of venience was conducted at the outpatient department of Ziauddin gestational diabetes. Hospital, Karachi from 1st July 2014 to 31st January 2015. The Impact of pharmacist intervention on antibiotic prescrip- study participants were 250 menopausal women between the tion orders in pediatric setting ages of 45–60 years. Data was collected by means of a ques- Uzma Shahid1, Mirza Tasawer Baig2, Ali Akbar Sial2and Zahida tionnaire, which consists of three parts containing questions about Memon1 demographic information, knowledge and attitude. 1Faculty of Medicine, Ziauddin University, Karichi, Pakistan,2Fa- Results: The mean age of respondents in the current study was culty of Pharmacy, Ziauddin University, Karachi, Pakistan 51.9 74.2 years. The mean age at menopause was 45.973.8 years. Introduction and objective: The prospective study was de- As far as the knowledge about menopause is concerned 4.8% had signed to formulate and test the efficacy and impact of trial in- weak, 71.2% had moderate and 24% had good knowledge about terventions by pharmacist on potential harm and incidence rate of menopause phenomenon. Majority of respondents had positive errant antibiotic medications dispensed in children admitted to a (72.4%) attitude towards menopause and 27.6% had negative atti- public sector hospital in Karachi. tude towards menopause. Methods: An eight months study was conducted in a teaching Conclusions: Appropriate knowledge and positive perception hospital comprising of 850 beds with 65 beds strength in three towards menopause is very important to help women to cope with pediatric units I, II and III inclusive of neonatal ICU. Orders written menopause transition and can help in the development of ap- by physician groups in all patient admission categories were in- propriate programmes to promote women health. cluded. The prescription order review prior to dispensing was Social support as a determinant of internalized stigma and made by experienced and specialized pharmacists in pediatric self-esteem in persons with schizophrenia pharmacotherapy in a research setting. Prescription order in error Umara Rauf1and Uzma Ali1 was marked if not in compliance with standard pediatric refer- 1Institute of Clinical , University of Karachi, Pakistan ences, dosing guidelines and antibiotic prescription guide adopted Introduction and objectives: Social support works as a buf- in the hospital. As per predefined criterion in our study, errors fering factor in psychotherapeutic treatment for the diagnosed were labeled as potentially lethal, serious and significant. The data persons with schizophrenia in reducing internalized stigma and in was statistically compared by unpaired samples test and the effi- boasting their self-esteem. The aim of the present study is to ex- cacy of the intervention on type of errors was margined by chi plore the predictive relationship of social support with inter- square test. nalized stigma and self-esteem among diagnosed persons with Results: 8.5 medication errors per 1000 prescription orders schizophrenia. were identified (1.5 medication error per 50 patient per day), Methods: A purposive sample of 52 diagnosed persons with where-as 18 potentially lethal errors were identified within the schizophrenia, age ranges 18–55 years who belong to different span of 8 months. Potential cardiopulmonary arrest and anaphy- socioeconomic status was taken from different psychiatric hospi- lactic risk was significantly assessed (po0.01) in ICU. Overdose tals of Karachi. Demographic Information sheet, Multi- and under dose of antibiotics accounted to 76% errors. The N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 193 frequency of errors was comparatively non significant in pre- Ahmed1 scription orders by senior attendees (po0.05). 1Faculty of Pharmacy, Jinnah University for Women, Karachi, Conclusions: Effective control of antibiotic medication errors is Pakistan observed with the integrated role of pharmacist in the prescrip- Introduction and objectives: Type 2 diabetes mellitus (T2DM), tion-dispensing-administration sequence. non-insulin dependent diabetes mellitus, is the most rapidly Simultaneous determination of six antihistamine anti- growing disease, estimated to affect 439million adults by 2030. allergic drugs in pharmaceutical formulations, human serum The rate is faster in developing countries than developed countries and pharmacokinetics application which in turn results in higher healthcare expenditure. Thus it Nawab Sher1*, Farhan Ahmed Siddiqui2, Najmul Hasan3and becomes a public health priority to prevent the disease and con- Nighat Shafi2 trol over its rapid growth. Among the known risk factors that 1Department of Chemistry, Faculty of Science, University of Kar- contribute to T2DM are dietary factors, which promote early dia- achi, Pakistan,2Faculty of Pharmacy, Federal Urdu University of Arts, betes in healthy individuals and increased mortality and morbidity Science and Technology, Karachi, Pakistan,3Department of Micro- in type 2 diabetic patients. To evaluate the hypothesis that fruits biology, Faculty of Science, University of Karachi, Pakistan provide more protection against T2DM than fruit juice, we per- This article describes a new, accurate and highly specific high formed experiment by giving fruits to one group and fruit juice to performance liquid chromatographic method with UV detection another group of healthy volunteers and monitor fasting blood (HPLC-UV) for the simultaneous determination of cetirizine HCl, sugar and random blood sugar. chlorphenamine maleate, loratadine, domperidone, buclizine and Results: The results of current study suggested that whole meclizine in pharmaceutical dosage form and human serum, in- grapefruit and orange fruit are very effective in maintaining ran- volving pyridoxine as internal standard. The mobile phase consists dom blood sugar levels in comparison to juices. of heptane sulphonic acid salt buffer and acetonitrile, drawn at a Results also suggested that dietary fibers are not only effective flow rate of 1.0 ml/min using a symmetry C18 column with UV in regulating blood sugar, but also showed insignificant change in detection at 230 nm. The intraday and inter-day precision mea- systolic and diastolic blood pressure and also in pulse rate. surements showed coefficients of variation always less than one. Moreover, whole grapefruit has been found more effective in The calibration curve was tested in the range of 10–2150 ng/ml and maintaining pulse rate in comparison to fibers obtained from ap- the correlation coefficient of 40.9990 in all cases was obtained. ple fruit and orange fruit. The averages of the absolute and relative recoveries were found to Conclusions: Based on above findings, it has been concluded be in the range of 98–102%. Up to six drugs were separated in the that patients with type 2 DM could be maintained on dietary fibers same chromatogram with good resolution. The proposed HPLC not only for better glycemic control, but also for controlling dia- method has reasonable applications in pharmaceutical tablet do- betes related complication such as high blood pressure. This study sage form and pharmacokinetics studies. could further be extended to assess the effects of dietary fibers Simulation based cardiopulmonary auscultation: a tool to (especially obtained from grapefruit) in controlling other diabetes aid medical students of Ziauddin University related complications, e.g. hypercholesterolemia, diabetic ne- Iqrah Iqbal1, Kulsoom Farooqi1and Fatima Z Mithani1 phropathy, in diabetic patients. 1Ziauddin University, Karachi, Pakistan NAFLD in diabetic patients treated on oral hypoglycemic Introduction and objectives: Use of simulation and state of art Dunesh Kumar1, Zahida Memon1and Samia Perwaiz1 mannequin and models in clinical skills lab is now becoming a 1Department of Pharmacology, Ziauddin University, Karachi, common practice which is offered to 3rd year medical students Pakistan during their clinical rotations. We seek to determine the efficacy of Introduction and objectives: Determination of NAFLD (non- cardiopulmonary auscultatory skills in 3rd year medical students alcoholic fatty liver disease) in diabetic patients on oral hypogly- of Ziauddin University who have been trained by simulation based cemic therapy. Diabetes is one of the most common non-com- auscultatory device. municable disorder throughout the world. It has enormous com- Methods: Students who participated in this study (n¼40) were plications, which indirectly multiply the economic and medical from 3rd year MBBS program. They were assigned in 2 equal burden. According to WHO, 9% population of the world suffers groups. Group A took a lecture on the detection and knowledge of from Diabetes who are above 18 years age. Prevalence of diabetes various cardiopulmonary sounds whereas Group B was tutored on in Pakistan is about 10% according to Akhtar. This poses a huge a simulation based device after they had taken their lecture. After burden on a country like Pakistan, which already has very limited respective training sessions, each group was tested with the help resources in field of health. of a questionnaire. This questionnaire consisted of identification Deficiency of Insulin causs hyperglycemia and hyperlipidemia. and knowledge based open ended and close ended questions. Data This leads to multiple disorders like atherosclerosis, CVAs, IHD, was entered into data analysis software S.P.S.S ver 15, Data was Renal failure, NAFLD, glaucoma, cataracts, gangrenes of limbs, analyzed and mean, standard deviation was calculated along with acute emergencies like DKA (diabetic ketoacidosis) and HONKC Charts, graphs were generated to represent results. (hyperosmolar non ketotic coma) etc. NAFLD (Non Alcoholic Fatty Results: Diagnostic accuracy and correct identification of car- Liver Disease) was first described by Ludwig in 1980 and is on rise, diopulmonary sounds was better in the group that had been as- making it the most common cause of CLD. Commonly called fatty sisted with a simulation based device. Mean score of assessment of liver diseases is a major problem arising in diabetic population, knowledge and skills of cardiopulmonary sounds were found which has been neglected off until now. The prevalence rises to significant in group B (mean ¼13.571.73) compared to group A 60.8% in diabetics in Pakistan. 40% cases of NAFLD can progress to (mean ¼6.972.64). NASH (nonalcoholic steatohepatitis) of whom 32–37% can progress Conclusions: According to our study at Ziauddin medical uni- to advanced fibrosis. This prevalence rises to 35–75% in obese and versity, it is suggested that simulation based learning is a sub- diabetics. Making it most common cause of cryptogenic cirrhosis, sidiary tool in preserving knowledge and skills for cardio- which can lead to hepatocellular carcinoma. NAFLD is usually an pulmonary auscultation. incidental finding when an abdominal scan is done for some other Whole fruit better than fruit juices in maintaining blood suspected pathology. sugar levels Methods: FBS, Fasting LIPID profile andHbA1C was done. Ul- Somia Gul1, Humera Khatoon1, Haneen Rashid1and Nameera trasound was done to see fatty infiltration of liver. 194 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

Conclusion: Ultrasound liver of type II diabetic patients factors showing non-alcohol fatty liver changes. Sarah Baber1, Sana Ilyas1and M Ammar Hassan1 Identifying errors in prescription writing 1Ziauddin University, Karachi, Pakistan S H Afsheen Nazar1, Nasim Karim1and Riffat Farooqui1 Introduction and objectives: In any population, women are 1Department of Pharmacology and Therapeutics, Bahria University more prone to depression. In our middle class community it is still Medical and Dental College, Karachi, Pakistan considered a taboo. Housewives usually emphasize on somatic Introduction and objective: The errors of prescribing are the issues rather than emotional signs and symptoms which causes commonest form of avoidable medication errors and are con- detrimental effects on their health. Timely identification of risk sidered to be the most important target for improvement. Medi- factors and counseling of women regarding their mental health is cation errors are common in general practice. Both errors in the of utmost importance. We aim to measure frequency and identify act of writing (prescription errors) and prescribing faults due to major risk factors of depression in housewives belonging to mid- erroneous medical decisions can result in harm to patients. Our dle class community in Karachi. objective is to identify the errors in the prescriptions of general Methods: Cross sectional study conducted on housewives be- practitioners (GPx) from different parts of Karachi. tween 25 and 50 years of age. Through convenience sampling Methods: A descriptive pilot study was conducted from 1st technique 247 questionnaires with 17 questions were adminis- January to 30th February 2014. A total of 100 prescriptions were tered, followed by the patient’s health questionnaire 9 (PHQ-9). collected from 4 general practitioners’clinics (east, west, central Data was analyzed by SPSS v20. Chi-test was calculated for cate- and south districts) of Karachi. Verbal consent was taken few days gorical data and p-value o0.05 was considered significant. prior to random collection. All prescriptions were analyzed for Results: Out of all the respondents, 131 females (53%) turned errors in superscription, inscription, subscription, transcription, out to be depressed and 116 (57%) not depressed. PHQ-9 classifi- signatures and refill information. cation of the levels of depression in the participants showed 100 Results: A total of 373 errors were identified in 100 prescrip- females mildly depressed, 77 females moderately depressed, 35 tions. 203 errors in superscription, 6 in subscription, 34 in tran- females moderately-severely depressed, 16 females severely de- scription, 1 in prescribers’ signatures and 62 in refill information. pressed and 19 females not depressed. Conclusion: Errors in prescription writing are found to be Conclusions: Most prominent risk factors for depression were common in the prescriptions of general practitioners. Measures of having to take care of an old or an ill person at home and living should be taken to refresh the prescription writing skills of general in a joint family structure. Although the study targeted middle practitioners through CMEs and workshops. class community, the analysis showed no associations with the Assessment of stressors among undergraduate dental stu- family income. dents of Karachi, Pakistan Frequency and type of chewable tobacco commonly used by Safia Anwar1and Saira Hamid1 oral squamous cell carcinoma patients 1Bahria University Medical and Dental College, Karachi, Pakistan Fauzia Perveen1and Saeeda Baig1 Introduction and objectives: Dental schools are known to 1Department of Biochemistry, Ziauddin University, Karachi, have highly stressful learning environments. Dentistry involves an Pakistan acquisition of required academic, clinical and interpersonal skills Introduction: South East Asia is threatened by the risk of Oral during the course of learning. Identifying sources of stress re- Squamous Cell Carcinoma. The popularity of chewable tobacco presents the crucial first step towards advocating policy changes plays a pivotal role in making it the second leading malignancy in and enhance student’s stress coping skills. This study was con- Pakistan. Prolonged use of these products can lead to different oral ducted to assess the problems related to stress encountered by precancerous conditions, GI cancer, ulceration and many other dental students of Karachi. The aims are: (a) To identify the diseases. The objective of the study was to find out the frequency common stressors among undergraduate dental students of Kar- and type of chewable tobacco, which has been the cause of Oral achi. (b) To investigate whether specific stressor is related to year Squamous Cell Carcinoma in these patients. of study or gender. (c) To evaluate the factors which influence Methods: The study was descriptive. Seventy one patients who student perception that might allow programmatic changes. had Oral Squamous Cell Carcinoma and a history of tobacco use Methods: Type of Study: Cross Sectional Survey; Sample Size: were selected from cancer OPD. Subjects addicted to stuff other 330 undergraduate dental students of Karachi; Study population: than smoking and chewable tobacco were excluded. A ques- second, third and final year BDS; Study setting: Five dental schools tionnaire was used to collect demographic characteristics, oral of Karachi; Modified form of Dental Environment Stress ques- hygiene, medical history, family history of cancer, and detailed tionnaire was administered. history of tobacco usage, chewable or smoke products and details Results: A total of 500 Questionnaire were distributed out of of chewing habits (using since, packs/day, duration of exposure which only 330 responded. Of the respondent, 70% were women etc.). The study was carried out in Ziauddin. and 30% were men. The most stressful variable among all was Results: Majority of the subjects were males (80%). Out of the ‘Overloaded feeling due to vast syllabus’. Third and final year 71 Oral Squamous Cell Carcinoma patients majority 42 (59.15%) students were reported to have higher-level stress than second were multiple users that consumed more than one tobacco pro- year students. Females were more affected by the same stressor duct. Whereas, 9 (12.67%) consumed gutka, 9 (12.67%) pan, 5 (7%) than male students. betel nut, 4 (5.6%) niswar and 2 (2.8%) cigarette. Conclusions: The conclusion of study was drawn from vari- Conclusions: Frequency of OSCC was found higher in users of ables with significant values o0.005. Stress due to accommoda- multiple chewable tobacco formulations which makes consump- tion problem was higher in students of second year while clinical tion of multiple tobacco formulations one of the major etiological based problem caused greater level of stress among students of factors in the development of OSCC. third and final year. The variables relating to academics (in- Molecular journey of Vitamin D, VDR and VDRGP and its adequate break timings, inadequate time for assigned work and association with breast cancer overloaded due to vast syllabus) caused stress in all three years Samiuddin Ahmed1, Saiyed Abdullah Abubakar1and Bushra equally. Stress showed an upward trend from the first to the final Wasim1 year with a jump from the second to the third years. 1Ziauddin University, Karachi, Pakistan Frequency of depression in house wives and associated risk Vitamin D plays an essential role in regulating the levels of N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 195 calcium and phosphate in the body. Vitamin D is not active as a high Kþ-induced contractions. At 0.01 mg/ml, Mn. Cr caused a hormone - instead, a few chemical changes must be made. These parallel shift in acetylcholine (ACh) concentration-response curves are performed by enzymes that add hydroxyl groups. Vitamin D (CRCs) followed by a non-parallel shift at 0.03 mg/ml, similar to receptor is a nuclear hormone receptor Transcriptional regulator in dicyclomine. At further tested concentrations, Mn. Cr (0.1 and response to 1,25, dihydroxyvita-min D 3. 0.3 mg/ml) and fraction suppressed Ca2þ CRCs, similar VDR protein is at the center of the Vitamin D endocrine system to verapamil. In guinea-pig ileum, Mn. Cr, its aqueous and ethyl with a negative feedback which regulates serum calcium and Vi- acetate fractions exhibited atropine-sensitive gut stimulant activ- tamin D, if not maintained will affect large number of organs in- ity along with additional uncharacterized excitatory response in cluding breast. Data is available in positive genetic association of aqueous fraction only. certain diseases which suggest functional consequences of VDR Conclusions: These results suggest that black mulberry pos- gene polymorphism. Vitamin D plays a role in breast cancer and sesses prokinetic, laxative and antidiarrheal effects, putatively exerts its effect through VDR gene polymorphism. There are four mediated through cholinomimetic and anti-muscarinic plus polymorphic variants of VDR (Folk 1, Bsm1, Taq 1 and Appa1). Ca2þ-antagonist mechanisms, respectively. Hormone Reception-Vitamin D hormone binds to receptors in its Antibacterial activity of honey and turmeric against git and target cells, controlling the synthesis of many different proteins respiratory pthogens involved in calcium transport and utilization. The receptor is Hira Jalal1and Aliya Hayat1 composed of two domains: a domain that binds to the hormone 1Department of Microbiology, Jinnah University for Women, Kar- and a domain that binds to DNA. It pairs up with a similar protein, achi, Pakistan 9-cis retinoic acid receptor (RXR), and together they bind to the Introduction and objectives: Because of emergence of re- DNA, activating synthesis in some cases and repressing it in others. sistant pathogens the antibiotic activity is diminished. This poses a There is a decrease in VDR expression in breast cancer cells com- serious threat to public health. Therefore alternative antimicrobial pared to normal breast cells and this decrease could be due to techniques are needed. Thus this situation has led to reevaluation gene polymorphism and other factors also Alterations in VDR ex- of the therapeutic use of ancient remedies such as plants and plant pression and activity could lead degranulation of vitamin D up- based products including honey and turmeric. take, metabolism and serum levels of biologically active Vitamin D. Methods: The study was preformed to evaluate the anti- Potential of black mulberry: From food of choice to remedy microbial activity of honey and turmeric, purchased from local for bowel dysmotility market. Different concentrations of honey and turmeric were Malik H Mehmood1*, Anam Akhlaq1, Zohaib Ashraf1, Sadia Syed1, prepared in distilled water including 100%, 75% and 50%. Its effect Samira A Bawany1, Abdul Rehman1, Anwarul-Hassan Gilani1,2 was observed on isolates obtained from clinical samples, which 1Natural Products Research Division, Department of Biological and includes Salmonella typhi, Shigella dysentrey, S.aureus, Proteus Biomedical Sciences, Aga Khan University, Karachi, Pakistan,2Pakistan mirabilis, E.coli, Pseudomonas aerignosa, Klebsiella oxytoca, Ba- Council for Science and Technology, , Pakistan cillus subtilis, Enterococcus and Micrococcus leutus. The study was Introduction and objectives: Morus nigra Linn. (black mul- carried by disc diffusion method. All experiments were performed berry) is used in gastrointestinal ailments. This study demon- in tricplicate. Zone of inhibition was measured in millimeter and strates gut modulatory proper-ties of M. nigra. results were given in percentage. Methods: The prokinetic, laxative and antidiarrheal activities of Results: Honey extract show highest antibacterial activity as M. nigra were assessed in mice, while isolated rabbit jejunum and compared to turmeric extract. Highest concentration (100%) of guinea-pig ileum were used to explore insight into mechanism(s). honey and turmeric was inhibitory to all strains among which Results: At 30 and 70 mg/kg, the crude extract of M. nigra (Mn. Enterococcus gave 75% inhibition while other strains were in- Cr) exhibited atropine-sensitive prokinetic and laxative effects, hibited up to 50%. It was found that 75% concentration of these similar to carbachol (CCh). While at higher doses (100–500 mg/ compounds gave intermediate zone of inhibition while 50% con- kg), Mn. Cr offered protection against castor oil-induced diarrhea. centration was least inhibitory. In rabbit jejunum, Mn. Cr and its chloroform fraction inhibited Conclusions: Honey and turmeric have potential to use as CCh-induced contractions more potently compared to high Kþ therapeutic agents either alone or in combination against various (80 mM). Conversely, petroleum fraction was more potent against infections. 196 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

Abstracts: Joint 2nd Annual Conference of Ziauddin University, HEC Pakistan with European Society for Translational Medicine (EUSTM) International Conference on Translational Medicine: From Discovery to Health Care

Oral Presentations UNDERSTANDING TRANSLATIONAL MEDICINE DR. N.A. JAFAREY MEMORIAL LECTURE Aamir Shahzad1 CO-ADMINISTRATION OF TURMERIC AND KALONGI (BLACK 1European Society for Translational Medicine (EUSTM), Vienna, SEEDS) SHOWED ENHANCED EFFICACY IN METABOLIC DIS- Austria and School of Medicine, University of Colorado, Aurora, USA. ORDERS – FROM BENCH TO BED, TRANSLATIONAL STUDY In the recent past, Translational Medicine (TM) has gained Anwar ul Hasan Gilani1 significant importance due to its promising role for accelerating 1Pakistan Council for Science and Technology, G-5/2, Islamabad research and development in academia and industry. TM has great and Department of Biological and Biomedical Sciences, Aga Khan potential to enhance health care and better patient care starting University Medical College, Karachi, Pakistan. from prevention, diagnosis, and drug/device developments for Amongst non-communicable diseases, metabolic syndrome clinical disorders. Translational medicine tools and techniques (MS), a cluster of metabolic disorders including obesity, hy- help to plan and improve infrastructures and capabilities while perglycemia, dyslipidemia and hypertension, is highly prevalent in reducing cost and optimizing resources. This talk will provide an modern society. Due to its rising incidence and demanding life- overview of translational medicine field including definitions and long use of multiple drugs, there is a growing interest in testing concepts, techniques and tools, potential benefits as well as global and developing safe alternative reme-dies. initiatives and programs. Animal studies were conducted to establish a better rat model DIGITAL PATHOLOGY – AUTOMATED RECOGNITION OF CAN- of MS with low HDL. Replacing whole wheat with refined wheat CER CELLS FROM BODY FLUIDS flour in rat chow in 60 % fructose-fed Sprague-Dawley rats resulted Gottfried Koehler1 in hypertension, hyperinsulinemia, hyperglycemia and a reduction 1Max F. Perutz Laboratories, University of Vienna, Austria. in HDL levels at 4 weeks, while hypertriglyceridemia with en- The Talk will cover the full spectrum of topics currently making dothelial dysfunction was observed at 8 weeks. This rat model was an impact on the digital pathology practice. Whole slide imaging therefore, used to study the efficacy of co-administration of Tur- has the potential to improve efficiency and cost-effectiveness, meric and Kalongi on parameters of MS. The HPLC fingerprinting while providing access to high quality care in areas that are cur- of the crude methanolic extract of Turmeric showed the presence rently under-served. Many institutions are leading the way with of curcumin mainly with a few minor unidentified compounds the use of whole slides images for primary diagnosis, while others while, the extract of Kalongi seeds showed the presence of thy- are trailing behind. Progress being made in digitization and re- moquinone along with some other unidentified compounds. cognition and the advantages of taking strides to digitize in- At 3 weeks of the treatment of rats with Kalongi (03 g/kg) and formation to share images and data across institutions as well as Turmeric (1.5 g/kg) lowered raised blood pressure and low density encourage adoption by medical institutions and thereby lending lipoprotein (LDL), respectively, while their co-administration at expertise across geographic borders will be evaluated. Technical half the dose of individual herb reduced blood pressure and tri- progress in new technology, data analysis, machine learning and IT glycerides. At 6 weeks, higher dose of Kalongi alone (0.6 g/kg) infrastructure in addition to standardization of practice will be prevented hypertension, hyper-glycemia, dyslipidemia and en- highlighted with emphasis on non-invasive cancer diagnostics dothelial dysfunction but had no effect on cholesterol and serum based on liquid biopsies. insulin levels when compared with fructose-fed controls. Turmeric PHYTOMEDICINE FOR HEALTHCARE IN THE 21ST CENTURY alone, in a higher dose (3 g/kg) prevented dyslipidemia, but had no M Iqbal Choudhary1and Atta-ur-Rahman1 effect on blood pressure. Co-administration of half the doses of 1International Center for Chemical and Biological Sciences, H.E.J. individual herbs (Turmeric 1.5 g/kg and Kalongi 0.3 g/kg) offered Research Institute of Chemistry and Dr. Panjwani Center for Mole- better efficacy in wider range of parameters of MS such as pre- cular Medicine and Drug Research, University of Karachi, Pakistan. vention of hypertension, hyperglycemia, dyslipidemia, hyper- Global healthcare system is challenged by emerging diseases, insulinemia and endothelial dysfunction. equity issues, and most importantly sustainability. Nature pro- Study conducted on human revealed that around 30 % were vided perfect system to nature, sustain and improve life for human overweight with nearly 60 with central (abdominal) obesity and and other life forms since the beginning. Some time during the last when Kalonji and Turmeric given in combination with 60% dose of century, capacity to synthesize new molecules has made its way in the individual herbs, showed an improvement in all studied drug discovery and development. The mutual co-existence of both parameters of MS including in body fat, cholesterol (also HDL) and systems, i.e. nature-based healthcare and synthetic based medi- blood glucose. The results therefore, for the first time, revealed cines, could have been much better, but obsession of pharma- enhanced effect of a low dose combination of Turmeric and Ka- ceutical industries to quickly bring blockbuster drugs to market- longi on parameters of MS among animals as well as humans and place totally distorted the focus. Overemphasis on synthetic this study has huge impact on public health offering relative-ly medicines has not only seen the world’s indige-nous knowledge safe and cost-effective remedy for some chronic diseases. and practices diminished, but also marginalized and disowned the This study was part of PhD thesis of Dr. Faridah Amin graduated indigenous healthcare systems which supported the human race from Aga Khan University in 2015. for so long. Today there is a major disconnect between the two http://dx.doi.org/10.1016/j.nhtm.2016.11.001 2307-5023/& 2016 European Society for Translational Medicine. Published by Elsevier Ltd. All rights reserved. N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 197 systems, where nature-based drug discovery is grossly neglected. Results and Conclusion: Majority of the patients were happy Cost of bringing a new drug is extremely high (US $ ~ 2.0 billion), with the results. Muscle splitting biplane is a versatile pocket and leaving poorman and orphan diseases untreated. Intellectually the its quick learning curve and reproducibility has made it a good developing world has no role in the global drug discovery and option for augmentation mammoplasty, augmentation with mas- development process. Business of pharmaceutical development is topexy in both primary as well revision procedures. not-able to meet the needs of majority of human population. Ir- NUTRACEUTICALS: THE SCIENCE BEHIND THE MYTH onically interest in nature-based drugs has never diminished. Khalid Rahman1 More so inability of modern medicines to treat human diseases 1School of Pharmacy and Biomolecular Sciences, Liverpool John and their reported side effects created a niche for herbal-based Moores University, Liverpool, . drug discovery and development. Epidemiological studies have shown that diets rich in plant The phytomedicine too despite their general acceptability, foods can either reduce or prevent chronic diseases e.g. cardio- growing interest and promise, face great challenges, such as fast vascular disease. Medicinal plants and nutraceuticals usage can be disappearing indigenous knowledge, loss of habitat of medicinal a cost effective way of sustaining and improving general health plants, proof safety and efficacy, and biopiracy. The presentation especially in countries with limited health resources. The concept will be an analytical review of the importance of herbal and nat- of healthy ageing and personalised medicine is also becoming ural medicines in 21st century, along with some of the most en- important. We have investigated the role of garlic in the preven- during challenges this is facing. Unfortunately many of these tion of cardio-vascular disease and the role of tea in pro- challenges are not even discussed. moting neutrophil activity both in vitro and in vivo. Garlic has ADVANCED GLYCATION ENDPRODUCTS: POSSIBLE LINK BE- attract-ed considerable interest as a potential cardio-protective TWEEN DIABETES AND BREAST CANCER agent whilst green tea is reported to enhance the immune system. Nessar Ahmed1 Garlic was given to healthy volunteers and it displayed antioxidant 1School of Healthcare Science, Manchester Metropolitan Uni- properties as judged by an increase in total antioxidant status and versity, Manchester, United Kingdom. inhibition of LDL oxidation; further more garlic prevented platelet Diabetes mellitus affects over 382 million people worldwide aggregation by mechanisms which include an increase in cyclic and this number is expected to increase to 592 million by 2035. nucleotides, inhibition of fibrinogen binding and platelet shape Diabetes is characterized by hyperglycaemia, a consequence of change. Neutrophils play a defensive role against bacterial infec- which is increased protein glycation and the subsequent formation tions by oxygen dependent and independent pathways. Neu- of complex heterogeneous molecules called advanced glycation trophils were isolated from healthy volunteers and primed and endproducts (AGEs). Diabetic patients often have an increased activated with formyl peptide fMethionine-Leu-cine-Phenylala- susceptibility to certain types of cancers including breast cancer. nine (fMet-Leu-Phe) and lactoferrin and myeloperoxidase was Although AGEs have been implicated in many diabetic complica- measured as well as water soluble antioxidant status. Green tea tions, their effects on breast cancer cells are not well studied. This extract was then given to volunteers and the assays repeated; study investigates the effects of AGEs on the invasive MDA-MB- increase in lactoferrin, myoperoxidase and water soluble anti- 231 breast cancer cell line and showed that AGEs increased pro- oxidants was observed. In conclusion, garlic has the potential to liferation, migration and invasion of these cells in a dose-depen- reduce cardio-vascular disease whilst green tea may protect dent manner. Furthermore, AGEs also up-regulated the expression against bacterial infections and both garlic and green tea have the of the receptor for AGEs (RAGE) and of the key signalling mole- potential to protect against oxidative stress in healthy volunteers. cules. Blockage of AGE interaction with RAGE inhibited AGE-in- Nutraceuticals have an important role to play in the field of duced effects including signalling pathways in this breast cancer Translational Medicine. cell line. There-fore, AGEs might contribute to breast cancer de- TRANSORAL LASER APPLICATION FOR VOICE CONSERVATION velopment and progression in diabetes. Antiglycation compounds AFTER SURGERY FOR EARLY GLOTTIC CANCERS may have a role in treating diabetic patients with breast cancer. Gurmit K Bachher1 USE OF MUSCLE SPLITTING BIPLANE IN PRIMARY AND RE- 1Speech and Language Pathologist, Tata Memorial Hospital, VISIONARY BREAST AUGMENTATION AND AUGMENTATION Mumbai, India. WITH MASTOPEXY. Introduction: The surgical laser has significantly improved the Umar Daraz Khan1 ability of the head and neck surgeon to work within confined 1Aesthetic Plastic Surgeon, United Kingdom. spaces. It permits meticulous removal of cancerous tissues within Introduction: In Muscle Splitting Biplane technique, muscle the larynx without damage to adjacent tissues. lies in front and behind the implant at the same time. Submuscular All cancer of larynx patients were resected with CO2 Laser at positioning of the implant is achieved by splitting muscle along its Tata Memorial Hospital fibres direction without muscle release. The concept is used for The voice quality of these patients was assessed objectively to augmentation mammoplasty and augmentation mastopexy in find out the improvement in it. primary and secondary cases. Voice parameters assessed were: Methods: Since 2005, author performed 1670 implant related . The technique was used in primary augmentation mam- 1. Frequency Parameters moplasties (1298), primary augmentation mastopexies (108), 2. Intensity Parameters 3.Perturbation Characteristics 4.Vocal tract multiplane pocket for augmentation and internal glandulopexy Parameters through infra mammary crease (63). Muscle splitting biplane is also performed in patients requesting for change of implants fol- Results: Demography of Patients is as shown below: n = 2380 lowing partial submuscular and subglandular mammoplasties T1 Glottic Cancers All male patients (75), patients presenting with synmastia (6), bottoming down Median Age = 56 years (range 27 – 73 years) No adjuvant (26), capsular contracture (30), rippling of the breast (30), rupture treatment was offered to them. Evaluation of implants (9) and revision mastopexies following subglandular All patients evaluated more than 3 months after treatment. implant position (13). Author has also used the pocket to correct Vowel /ee/ sustained for 5 secs. dynamic deformity (12) seen following partial submuscular breast Patients explained, trained and later evaluated Microphone to augmentation. mouth distance of 3 inches Recording of speech and its analysis 198 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 using Dr.Speech Voice Evaluation Software ver. 5 therapies or a prophylactic/therapeutic vaccine. Previous studies SPSS ver. 10 was used for Statistical Analysis Method have shown that viral, genetic and immunological components of Conclusion Voice after laser resection of the vocal cord is these LTNPs play a role in containment of the virus. My pre- qualitatively affected. sentation will focus on a few studies from own group as well as Regular voice therapy is required to maintain the voice future projects with reference to HIV studies in Pakistan. characteristics. CURRENT STATUS OF TRANSLATIONAL RE- Patient needs to be educated in Vocal hygiene to avoid further SEARCH IN PAKISTAN damage to voice. Nikhat Ahmed Siddiqui1 CHILDHOOD ACUTE LYMPHOBLASTIC LEUKEMIA 1Dean Research, Ziauddin University, Karachi, Pakistan. Zehra Fadoo1 The enriched history of neuroscience research dates back to 1Paediatric Oncology Fellowship Program, Aga Khan University, 4000 B.C., when euphoriant effects of poppy plant were reported Karachi, Pakistan. in Sumerian records with the breakthrough of 52 discrete cortical Acute lymphoblastic leukemia (ALL) is the most common areas by Korbinian Brodmann in the 19h century. childhood cancer. It constitutes approximately 25% of cancers in The potential impacts of advancements on science, medicine, children below 15 and 19% in children below 20 years of age. industry and society are expected to be profound. Translational Several risk determining variables have been identified in child- neuroscience is a relatively new term in brain research focusing on hood ALL and include patient-specific, clinical, and genetic fea- a closer interaction between basic and clinical to tures. Children treated on current chemotherapy protocols have an expand understanding of brain structure, function and disease, event-free survival (EFS) that exceeds 85%. This success is mainly and translate this knowledge into clinical applications and novel due to treatment stratification according to the risk of relapse and therapies of nervous system disorders. provision of better supportive care. It is essential to know the Pakistan being a developing country is much behind in brain disease characteristics and prognostic criteria to stratify treatment. research as compared to its neighboring countries. There is a MUTATIONAL LANDSCAPE OF HEAD AND NECK SQUAMOUS pressing need to establish a national body that encompasses all CELL CARCINOMAS IN PAKISTANI POPULATION types of neurosciences including behavioral, cognitive, computa- Kulsoom Ghias1 tional, molecular, neural aging, neuroanatomy, neurobiology, 1Biological and Biomedical Sciences, Aga Khan University, Karachi, neurochemistry, neuroendocrinology, neuropathology, neuro- Pakistan. pharmacology, neurophysiology and systems neuroscience. There Introduction: Head and Neck Squamous Cell Carcinoma is also a need for a large neuroscience center in Pakistan which can (HNSCC) is the most commonly diagnosed cancer in males and the train students in the neurosciences at the undergraduate and second most common in females in Pakistan. The increasing bur- graduate level. den of HNSCC in the region along with a unique set of risk factors Unfortunately, in Pakistan as in most parts of the world, mental merits a deeper investigation of the disease at the genomic level. health and mental disorders are not accorded the same degree of Methods: To this end, we performed whole exome sequencing importance as physical health. Rather, they are and have been of HNSCC treatment-naïve, HPV-negative cases and matched largely ignored or neglected. One of our projects on translational normal genomic DNA (n=7). neuroscience is based on revealing differential expression of nu- Results: We identified previously reported recurrent mutations clear proteins in Alzheimer’s disease and Schizophrenic’s human in TP53. Novel recurrent mutations in HNSCC in the Pakistani brain. We end up with discovering four major proteins to be dif- population have been provisionally identified in several genes, feren-tially expressed in AD brain including histone H4, H2B1B, including SERPINA3, CLMN, CHEK2 and PAK2. Using the IntOGen- TUBB4B and VDAC1. Further studies are needed to confirm this Mutations platform, we identified cell cycle and MAP kinase re- aberration and develop novel therapeutic targets for AD. lated pathways among the most commonly affected pathways. MENOPAUSE BOON OR BANE Conclusion: This data will be validated in a larger cohort and Rubina Hussain1 following functional analysis, the strengthened mechanistic 1Gynaecolgy and Obstetrics, Ziauddin University, Karachi, knowledge could contribute new, rational therapeutic targets for Pakistan. the treatment of HNSCC. Information regarding menopause is constantly evolving. Re- LONG TERM NON PROGRESSORS: AN EXCELLENT MODEL TO cent review of literature is being presented. Studies conducted in STUDY NATURAL HIV CONTROLLING MECHANISM FOR Ziauddin University showed majority of the young women are THERAPEUTICS aware of osteoporosis, the most common long term health im- Aneela Javed1 plication of menopause. It also showed that the main sources of 1Health Care Biotechnology, NUST, Islamabad, Pakistan. awareness in Pakistan are doctors whereas in most countries the Infection with the immunodeficiency virus (HIV-1) leads to the mass media and families are the main source. There is need for development of the acquired immunodeficiency syndrome (AIDS) Health & proper intervention programs. in humans that finally causes the death of the patient. To date, HIV A large number of women live half of their lives in a hormone lacks a suitable vaccine as almost all human HIV vaccine trials have deficient state without knowing how to alleviate this midlife invariably failed in vivo. Emphasis is now shifting to explore the phenomenon. The duties of health care professionals concerning natural mechanisms of HIV control that has been previously well menopause should be supportive, informative and interactive detected in long term non-progressors. In contrast to normal where women feel free to share their anxieties. Menopausal progressors (AIDS patients), approximately 5-8% of all HIV infected symptoms adversely affect quality of life. Hormone Replacement people fall into the group of long-term non progressors (LTNPs) or Therapy (HRT) is the most effective treatment. In spite of this, the Elite controllers (EC). These individual are infected with the virus risk- benefit ratio has always been debated. There is reduction in for more than 10 -20 years but do not show clinical symptoms of its use followed the report of Women’s Health Initiative. In- AIDS. Thus they maintain a very low viral load by certain natural dependent research shows that the continued negative attitude to mechanisms that are still not fully under-stood. LTNPs serve as an the use of HRT for symptom relief is not justified. Treatment ideal model for HIV-1 vaccine development due to their natural should be an individual decision of women after correct in- control of HIV-1 infection, since an understanding of their ability formation and advice from healthcare professionals has been to control HIV infection may lead to the development of immune given. N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 199

RELATION OF THYROID CANCER AND ANTHROPOMETRIC metabolic pathways, which will provide promise for future ther- FACTORS IN PAKISTAN apeutic developments in the treatment of schizophrenia. Sohail Awan1*, Adnan Ali1, Ramsha Shakeel2, Sadia Wasay2 and ANALYSES OF PRE-MRNA SPLICING MACHINERY BY UV-IN- Ahmed Rahim1 DUCED PROTEIN-RNA CROSS-LINKING AND MASS 1The Aga Khan University Hospital, Karachi, Pakistan, 2Dow SPECTROMETRY University of Health Sciences, Karachi, Pakistan. Uzma Zaman1and Henning Urlaub1 Introduction and Objective: We have investigated the re- Bioanalytical Mass Spectrometry Group, Max Planck Institute for lationship of thyroid cancer (TC) and anthropometric factors in Biophysical Chemistry, Goettingen, Germany. Pakistani population as this association remains to be Introduction and Objectives: The human genes that encode controversial. proteins undergo pre-mRNA splicing, an essential step of gene Methods: A total of 172 TC patients admitted at Aga Khan expression in which noncoding sequences are removed and coding University Hospital (AKUH) from 2000-2014 were included. Pa- sequences are ligated together. Unraveling splicing mechanism at tient’s demographics, tumor subtypes, body weight, height, body the molecular level is not only important for understanding gene mass index (BMI), body surface area (BSA) and body fat percentage expression, but it is also of medical relevance, as aberrant pre- (BF%) were calculated. mRNA splicing is the basis of many human diseases or contributes Results and Discussion: Out of 221 patients, 172 patients were to their severity. Although methods based on high-throughput included in this study . Mean age was 43 years ± 15.40. Among all sequencing have advanced our ability to identify the specificRNAs subtypes of TC, papillary remains the most common (72.1%), fol- bound by a particular protein, there is a need for precise and lowed by medullary (10.5%), follicular (7.6%) and others (9.9%). systematic ways to identify RNA interaction sites on proteins. Excess BMI 423 kg/m2, BF%, BSA, weight and height were sig- Methods: We have developed an experimental workflow nificantly associated with TC incidence in overall population (p- combining UV-induced cross-linking and high-resolution mass value o0.01*). In women, BF% was found to be in obese range spectrometry for the identification of protein-RNA cross-linking (mean=35.07% ± 7.6, p-value= o0.01*). In men, height 4160cm sites. Purified complexes were UV irradiated at 254nm and pro- and weight 460kg were positively associated with TC. No sig- teins therein hydrolyzed with trypsin. Peptide-RNA cross-links nificant relation was seen between gender and BMI. were enriched by TiO2 chromatography and analyzed by ESI-MS. Conclusions: Excess BMI 423kg/m2 and age 440years were Data analysis for identification of putative cross-links was per- positively associated with TC. In women, increased BF% and in formed by novel RNPxl pipeline. men, height 4160cm and weight 460kg had a significant rela- Results: The workflow not only enables the systematic identi- tion with TC incidence. fication of cross-linked peptides but also amino-acid residues in EXPRESSION ALTERATIONS OF METABOLIC PROTEINS IN RNA-interacting domains of proteins. SCHIZOPHRENIC BRAIN REGIONS Conclusions: Successful identification of several cross-linking Ayesha Khan1, Beena Hasan1, Shamim Mushtaq2, Abdul R Asif3and sites from human and yeast splicing complexes make it a pro- Nikhat Ahmed1,2 mising technique to study any RNA-protein complex of interest. 1Department of Biochemistry, University of Karachi, Karachi, CROSSTALK BETWEEN CYTOSKELETAL AND METABOLIC Pakistan,2Department of Biochemistry, Ziauddin University, Karachi, PROTEINS: A ROADMAP TO AD PATHOLOGY Pakistan,3University Medical Center Goettingen, Goettingen, Beena Hasan1,2, Ayesha Khan1, Christof Lenz3, Abdul R Asif3and Germany. Nikhat Ahmed4* Introduction: Schizophrenia (SZ), a multifactorial disorder 1Neurochemistry Research Laboratory, Department of Biochem- exhibits diverse neuropathological aberrations including defective istry, University of Karachi, Pakistan,2The Karachi Institute of Bio- neurotransmission system, neuroanatomical abnormalities and technology and Genetic Engineering (KIBGE), University of Karachi, impaired synaptic connectivity. Along with these complications Pakistan,3Institute of Clinical Chemistry, University Medical Center there is surfacing evidence that metabolic dysfunction and energy Goettingen, Germany,4Research Department, Ziauddin University, deficit may also contribute to its pathophysiology. Karachi, Pakistan. Methods: Present study entails, differential proteomic analysis Introduction and Objectives: Cellular activities are based on of autopsied schizophrenic brain substantia nigra, cortex, and complex networks of protein interactions. Identification and in- hippocampus (n=7) by using two dimensional electrophoresis teraction analysis of associated protein partners is the basis of (2DE) followed by Orbitrap mass spectrometry identification. many biological studies, determining the key etiological factors. Differential expression was validated by western blot, while In Alzheimer’s disease, a pathologically complex and multifactorial Silico analysis was applied for functional interactive role in disease is the most common cause of dementia. It is characterized metabolism. by a progressive decline in memory and cognitive functions. Results: Proteome mapping of SZ patients revealed more pro- Aberration of multiple pathways is observed in Alzheimer’s dis- found alterations in SN protein expression than other brain re- ease including metabolic pathways which subsequently alter the gions compared to controls. SN exhibits differential expression of allied cytoskeletal system. This study is designed to unveil the six proteins, enriched in several energy metabolic pathways in- cytoskeleton and metabolic proteins interaction in AD and inter- cluding decreased expression of phosphoglyceratemutase 1, ATP pret their association. synthase subunit d, mitochondrial and malate dehydrogenase cy- Methods: Human autopsied brain tissue samples from normal toplasmic, while increased expression of glyceraldehyde-3-phos- age matched control (n=6) and AD patients (n=5) were procured. phate dehydrogenase, 4-trime-thylaminobutyraldehyde dehy- Two dimensional Blue Native/SDS PAGE combined with Orbitrap drogenase, and alcohol dehydrogenase were found as compare to mass spectrometry analysis was accomplished to determine the normal brain. Cortex and hippocampus also portray significant interacting cytoskeleton and metabolic proteins. The interaction variations in expression patterns of five proteins across brain of was further confirmed by co-immunoprecipitation. schizophrenia patients. Results: A total of thirteen protein complexes were obtained on Conclusions: Identification of differentially expressed meta- BN-PAGE which resolved into many crucial proteins of different bolic proteins provides an insight for a better understanding of cellular networks. An important observation was a complex com- schizophrenia pathophysiology. Further characterization, will prised of novel interacting partners including glyceral-dehyde-3- elucidate the functional mechanisms underlying defective PO4 dehydrogenase, actin cytoplasmic, microtubule associated 200 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 protein 1B and glial fibrillary acidic protein. Further the interaction compared to control A and B (p-value=0.001). However control B between actin and GAPDH confirmed by co-immunoprecipitation showed significant gene expression as compared to control A (p- has not been established before in AD. value=0.001). The 2^^ct formula was applied to compare the fold Conclusion: In accordance with the previous findings of chain between the groups. The cases were also divided into ER, PR GAPDH and actin differential expression and post translational and HER2 positive and negative cases. It was observed that triple modification, our results suggest interesting implications of these negative cases showed a greater expression of gene as compared two proteins in AD. to other groups (p-value=0.001). DETECTION OF HIGH RISK HUMAN PAPILLOMA VIRUS (HPV) Conclusion: High expression of miR-21 in breast cancer pa- GENOTYPES 16/18 IN ORAL LESIONS OF TOBACCO CHEWERS IN tients suffering from stage III invasive ductal carcinoma had been PAKISTAN calculated as compared to its age matched healthy subjects. This Zile Rubab1*, Saeeda Baig1and Mohammad Haris Lucky1 result can mark miR-21 as a potentially strong diagnostic and 1Department of Biochemistry, Ziauddin Medical College, Ziauddin prognostic biomarker of breast cancer. MiR-21 profile was also University, Karachi, Pakistan. studied in daughters of the index cases. Their expression was also Introduction and Objective: The Human Papilloma Virus shown to be significantly higher as compared to the healthy in- (HPV) has evolved as a new culprit of malignant and premalignant dividuals but lesser than the full blown disease of breast cancer. oral lesions. The objective of this study was to find out the fre- This result strengthens the concept of inheritability of this disease quency of HPV and its high risk genotypes in different lesions of and this gene can also be labeled as a predictive biomarker. When oral cavity of tobacco chewers. Ct-values and fold change calculations were compared between Methods: From 492 subjects (421 males and 71 females), 20 ml ER, PR and HER2, higher expression was seen in negative tumor of oral rinse sample was collected after obtaining an informed tissues which are very resistant tumors and respond poorly to consent. Normal subjects with no chewing habits (250) including chemotherapy. 135 males and 115 females were also taken from same setting. SERUM METABOLOMIC PROFILES FOR BREAST CANCER DI- Gentle brushings over the lesions with the help of dental floss AGNOSIS, GRADING AND STAGING BY GAS CHROMATOGRAPHY- brush and stored at 4°C until DNA extraction. DNA was extracted MASS SPECTROMETRY and PCR was performed using HPV consensus primers Gp5+/Gp6+ Naila I Hadi1*, Qamar Jamal1, Ayesha Iqbal2, Fouzia Shaikh1, and HPV 16, 18 specific primers for genotyping. Categorical data Saleem Somroo3and Syed G Musharraf2,4 was calculated as frequencies and percentages. 1Department of Pathology, Ziauddin University, Karachi, Results: Oral pre-malignant lesions were present in 421 (86%) Pakistan,2Dr. Panjwani Center for Molecular Medicine and Drug Re- males and 71 (14%) females having leukoplakia (173, 35%), ery- search, International Center for Chemical and Biological Sciences, throplakia (60, 12%), submucous fibrosis (192, 39%) and L/E (67, University of Karachi, Karachi, Pakistan,3Breast Clinic, Surgical Ward 14%).Total number of HPV positives were 128 (26%), having HPV 16 2, Jinnah Postgraduate Medical Center (JPMC), Karachi, Pakistan,4H.E. (13%) and HPV 18 (11%) whereas, 76% had other genotypes. Among J. Research Institute of Chemistry, International Center for Chemical submucous fibrosis 82 (46%) were HPV positive. Out of total 128 and Biological Sciences, University of Karachi, Karachi, Pakistan. HPV patients 92% were males and 8% were females. All controls Introduction and Objectives: Detection of metabolic signature were found to be HPV negative. for breast cancer (BC) has the potential to improve patient prog- Conclusion: Frequency of HPV was found high (26%) in oral nosis. This study identified potentially significant metabolites lesions with HPV16/18 as 13% and 11% respectively. The patients differentiating between breast cancer patients and healthy con- with submucous fibrosis are at greater risk of having HPV. Other trols to help in diagnosis, grading, staging and determination of HPV genotypes causing premalignant lesions require further neoadjuvant status. investigation. Methods: Serum was collected from 152 pre-operative breast DIAGNOSTIC, PROGNOSTIC AND PREDICTIVE VALUE OF MI- cancer (BC) patients and 155 healthy controls in this case-control CRORNA-21 IN BREAST CANCER PATIENTS, THEIR DAUGHTERS study. Gas chromatography-mass spectrometry (GC-MS) was used AND HEALTHY INDIVIDUALS to obtain metabolic profiles followed by chemometric analysis. Ambreen Usmani1, Amir Ali Shoro1, Zahida Memon1and Mehvish Results: Chemometric analysis showed the identification of Hussain1 significantly differentiated metabolites including 7 for diagnosis, 1Bharia University and Ziauddin University, Karachi, Pakistan. 19 for grading, 23 for staging, 15 for stage III subcategories and 10 Introduction: MicroRNA-21 (miR-21) located on 17q23.1 has for neoadjuvant status (p value o 0.05). Partial Least Square shown high expression in breast cancer. This gene has anti- Discriminant Analysis model revealed a distinct separation be- apoptotic ability and causes tumor cell growth. It is also normally tween healthy controls and BC patients with a sensitivity of 80% involved in functions such as signal transduction pathways ef- and specificity of 100% on external validation. Models for grading, fecting normal cell growth and differentiation. staging and neoadjuvant status were built with Decision Tree Al- Objective: To determine the presence of miR-21 in the serum gorithm with predictive accuracy of 71.5%, 71.3% and 79.8% levels of breast cancer patients, their daughters and healthy respectively. individuals. Conclusion: Pathway analysis revealed increased glycolysis, Methods: A total of 132 subjects were recruited: 50 breast lipogenesis, and production of volatile organic metabolites in- cancer (cases), 50 age-matched healthy individuals (control A) and dicating the metabolic alterations in breast cancer. 32 daughters of index cases (control B). The sampling technique GROWTH FACTORS MEDIATED PRECONDITIONING OF ME- used for cases was random sampling. Controls were selected by SENCHYMAL STEM CELLS: STRATEGY FOR ENHANCING RENAL purposive method. Serum tests were run on qRT PCR and TISSUE REGENERATION threshold cycle was determined and fold change calculated. Rakhshinda Habib1,2, Kanwal Haneef1, Nadia Naeem1, Nurul Results: Fisher exact test was used when analyzing between Kabir1, Atta-ur-Rahman1and Asmat Salim1 cancer patients and healthy patients or with their daughters. 1Dr. Panjwani Center for Molecular Medicine and Drug Research, Normality of continuous variables was assessed by Shapiro Wilk’s International Center for Chemical and Biological Sciences, University test. While comparing the same between two groups, student t- of Karachi, Karachi, Pakistan,2Dow Research Institute of Biotechnol- test and Mann-Whitney test was used. P-value ≤ 0.05 was con- ogy and Biomedical Sciences, Dow University of Health Sciences, sidered significant. MiR-21 was significantly higher in cases as Karachi, Pakistan. N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 201

Introduction and Objectives: Acute kidney injury (AKI) is a mean of actual age (38.44±13.22) was not significantly different complex disorder manifested by rapid loss of renal function. AKI from the mean of calibrated age (37.52±12.36) which is estimated has strong influence on expression of renal specific genes which by regression equation (P-value=0.222 ). The mean error (differ- may play central roles in key cellular pathways and targeting these ence between actual and calibrated age) is found to be 0.911±7.41. pathways may turn out to be effective therapeutic option. Trans- Conclusions: The correlation coefficient was strongly positive. plantation of mesenchymal stem cells (MSCs) has been shown to The results strongly suggests that aspartic acid racemisation of improve kidney function following injury. However, poor survival human dentine is a precise method for estimation of chronological and grafting of stem cells to the site of injury has restricted their age in living and dead. The methodology should be standardized therapeutic efficacy. Preconditioning of stem cells can be a po- to make the results more accurate and prevent fallacies. However, tential strategy; chemokines and growth factors can improve their it remains an expensive procedure, considering that the laboratory potential in terms of homing and differentiation. Objectives of the equipment is not available in most cities of Pakistan. present study were to examine the changes in the expression CORRELATING VITAMIN D DEFICIENCY WITH VOLUMETRIC profile of genes associated with renal injury and to explore the BREAST DENSITY VIA A FULLY AUTOMATED SOFT WARE novel strategy of preconditioning of MSCs for injured kidney cells. VOLPARATMIN THE REPRODUCTIVE AGE GROUP Methods: Rat AKI model was established with gentamicin and Bushra Wasim1, Khalid Khan1and Farah Habib1 renal specific genes were analyzed through RT-PCR. For pre- 1Department of Anatomy, Ziauddin University, Karachi, Pakistan. conditioning strategy we used IL-7 gene to transfect MSCs. MSCs Objective: To see the correlation of volumetric breast density were co-cultured with cisplatin-treated injured Mardin-Darby and Vitamin D deficiency in the reproductive age group. bovine kidney (MDBK) cells and their in vitro fusion potential was Methods: We enrolled 300 patients from the general surgery analyzed. outpatient department, their blood samples were collected and Results: Several genetic changes occur in AKI that may con- vitamin D level was determined via chemiluminescence Method. tribute in long term renal injury consequences and targeting these Full field digital mammography was done and the DICOM images pathways may appear to be effective therapeutic options. Flow- were tagged and processed via fully automated software cytometry of preconditioned MSCs and injured MDBK cells re- VolparaTM by using the formula “Volumetric breast Density=100 x vealed significant (P≤0.001) cell fusion compared to that of the Volume of fibroglandular tissue (cm3)/ Volume of breast tissue normal MSCs. In addition, we also observed improved migration (cm3). ability of preconditioned MSCs in the in vitro wound healing assay. Results: There is enough evidence now that Vitamin D defi- Conclusions: For efficient stem cell therapy, it would be valu- ciency is inversely related to mammographic density and MD itself able to determine the contribution of various cellular factors in the is a strong predictor of breast cancer risk. Measurement of volu- fusion process so that a rationale for the use of such precondi- metric breast density objectively via a fully automated software tioned cells can be determined. “Volpara" and using the Raw Processed version of the FFDM in this AMINO ACID RACEMIZATION IN HUMAN DENTINE AS AN study was attempted for the first time in Pakistan and correlating INDICATOR OF CHRONOLOGICAL AGE – A STUDY IN KARACHI, with the vitamin D deficiency and also BI-RADS (Breast-Imaging PAKISTAN reporting and Data system) which is a common method used in Qudsia Hassan1, Allah Rakha Aswad1, Zahid Bashir1and Tehreem clinical practice worldwide. The total number of patients had only Ashraf1 two women with normal vitamin D level, the deficient group was 1Forensic Medicine Ziauddin University, Karachi, Pakistan. evaluated. In the group of women with low Vitamin D (Vitamin D Introduction and Objectives: Amino acid racemisation is a o 50), 76% had elevated VBD and 24% had normal VBD. The test reliable method to estimate age in developed countries. This study for proportions compares whether these two groups are sig- was designed to determine the coefficient of amino acid racemi- nificantly different in size or not. sation (AAR) with reference to age in our population. The objec- Conclusions: The null hypothesis assumes that vitamin D de- tives are to determine the coefficient of aspartic acid racemisation ficient women have the same number of people in tolerable and (AAR) with reference to age by HPLC, to measure the mean error in excess groups. Because the p-value is small, the null hypothesis is calculating age using this method; and to identify any differences rejected and we conclude that the number of women with ele- from earlier observation and discuss the reproducibility in our vated breast density is not the same as the number of women with setting. normal breast density, it is higher. Methods: Hundred (100) teeth were obtained from subjects NATURAL COMPOUNDS SUPPRESS OXIDATIVE STRESS AND between the age of 11 and 70 years who had come for routine INFLAMMATORY RESPONSE IN ACTIVATED MOUSE extraction procedure or for orthodontic procedures. Incisors, ca- MACROPHAGES nines, premolars and, first and second molars were included. Samreen Soomro1,3, M Ahmed Mesaik1, Farzana Shaheen2, Zaheer Carious tooth and third molar were excluded. Samples were Ul-Haq1and M Iqbal Chaudhary1,2 stored, dried, de-mineralized, hydrolyzed, and derivatised. High- 1Dr. Panjwani Center for Molecular Medicine and Drug Research, Pressure Liquid Chromatography (HPLC) was performed to quan- University of Karachi, Karachi, Pakistan,2H.E.J. Research Institute of tify the L- and D-forms of aspartic acid in dentin. Correlation and Chemistry, International Center for Chemical and Biological Sciences, regression was then tabulated based on the quantification, and University of Karachi, Karachi, Pakistan,3Dow Research Institute of was compared with data from other studies. Biotechnology and Biomedical Sciences DUHS, Karachi, Pakistan. Results: It was observed that there is very strong positive Introduction: Inflammation is quite complex, initiated by correlation (0.818) observed between actual age and co-efficient of several factors includes, molecules ranges from bacteria to che- racemization (statistically significant at 5%), which indicates that mical causing cellular trauma. Tissue injury induced by this trau- as age increase co-efficient of racemization increases. Based on the ma results in the inflammatory mediators release by macro-pha- analysis of 85 samples to estimate age least Square method was ges, including reactive oxygen species (ROS) like superoxide anion used to derived the regression line i.e. Age= -4.391 +347.396 (co- (O2-), hydrogen peroxide (H2O2), nitric oxide and increased ex- efficient of Racemization). Co-efficient of determination was found pression of pro-inflammatory cytokines, NF kappa B, and inducible to be 0.74, which show that regression equation for estimating age Nitric Oxide Synthase. A current study has led to the isolation of was 74% correct. The computed regression line was tested by using two new Acylphloroglucinols derivatives along with four known coefficient of racemisation for 15 samples, we observed that the compounds, Myricetin, Isousnic acid, G3-factor and 202 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 myrtocommulone E from Myrthus. disease. Intrapertum fever was recorded in 87% cases of EONS Methods: In this study, we used Lipopolysaccharide induced among which, 22% culture positive neonates were initially macrophages to study immune mediators release in culture su- asymptomatic. Apnea, fever and metabolic acidosis were the more pernatant. We also sought to further delineate the underlying prevalent symptoms. CBC, C-reative protein, Glucose and Blood mechanisms elicited by oxidative stress and inflammation and culture test were recorded. suppressed by compounds. Results: The most common pathogen was Group B streptococci Results: The results show that new Acylphloroglucinols deri- (24%), Klebsiella pneumonia (21%) and E.coli (15%). Fetal hypoxia vatives and Myricetin significantly suppressed the oxidative stress was confirmed in 34% cases. Ampicillin and Gentamycin was given and anti-inflammatory efficacy in suppressing, total reactive spe- empirically in 87% cases. Modified Kirby Baur disc diffusion tech- cies (ROS), superoxide, nitric oxide, pro-inflammatory inducible nique was used to determine antibiotic resistance. Klabsiella nitric oxide synthase (iNOS), proliferation of T-cells and nuclear pneumonia was greatly resistant against cephalosporins (average factor-kappa B (NF-κB) nuclear translocation. All the compounds of 54%) and co-trimoxazole (87.9%) whereas comparatively sus- were also non toxic to other cell systems. ceptible to quinolones (ciprofloxacin). Resistance of amikacin was Conclusion: Based on these results, we conclude that natural elucidated in 87% cases. product may contribute in anti-inflammatory drug discovery. Conclusion: In lieu with preventive approaches to avoid neo- IMIPENEM RESISTANT PSEUDOMONAS AERUGINOSA: THE natal sepsis, careful treatment choice of antibiotic is required to FALL OF THE FINAL QUARTERBACK deplete chances of therapeutic failure. Nadya Ameen1, Zahida Memon1, Shehla Shaheen1, Ghulam WITHANIA COAGULANS: A POTENTIAL CHOLESTEROL LOW- Fatima1and Farah Ahmed1 ERING PLANT 1Ziauddin University Karachi, Karachi, Pakistan. Tooba Lateef1, Sadaf Naeem1and Shamim A Qureshi1 Introduction and Objectives: To isolate, determine the fre- 1Department of Biochemistry, University of Karachi, Karachi, quency, and study the demographic trends of MBL positive Pseu- Pakistan. domonas aeruginosa from imipenem resistant isolates collected Introduction and Objective: Withania coagulans has many from clinical samples in a tertiary care hospital of Pakistan. medicinal properties. Currently its cholesterol lowering effect was Methods: In this cross sectional study a total of 230 strains of investigated in terms of HMG-CoA reductase inhibiting activity. Pseudomonas were isolated from various clinical specimens on the Methods: The study was designed to investigate in-vivo effect basis of culture and biochemical tests. Imipenem resistant isolates of methanolic fruits extract (MFEt) of W. coagulans in hyperlipi- were selected by Kirby Bauer Diffusion technique, followed by demic rabbits. On the completion of trial, total cholesterol (TC) and screening for MBL production by Imipenem EDTA Combined Disk HMG-CoA/mevalonate ratio were estimated. Additionally, in-silico Test. Demographic details of each patient were recorded on a se- study was performed to find out the inhibitory effect of 21 che- parate questionnaire. Chi-Square goodness-of-fit test was com- mical compounds present in fruits of same plant on HMG-CoA puted to review the isolation of MBL positive isolates (P-value ≤ reductase (HMGCR). Molecular docking calculations were carried 0.05) in different specimen. out by using Molegro Virtual Docker (MVD) software. Results: Out of 230 strains of P. aeruginosa 49.5% were imipe- Result: All doses of MFEt were significantly (po0.05) de- nem resistant; MBL production was confirmed in 64.9% of the creased TC and improved HMG-CoA/mevalonate ratio indicating resistant isolates. Resistance to polymyxin B (12.5%) was notable. HMGCR inhibition. Interestingly, 5 compounds were displayed the Majority of the MBL positive strains were isolated from patients highest MolDock score including Ergosta-5, 25-diene-3 β,24ε-diol, aged between 20-39 years (45.9%) and the predominant source Withacoagulin, Withanolide D, Coagulin D and Withaferin. Of was pus (43.24%) which was found to be statistically significant (P- which, excluding Withanolide D rest of the 4 compounds inter- value=0.04). Outpatient departments (24.3%) and burn unit (21.6%) acted with catalytic residues present in the active site of HMGCR were the major places for resistant isolates. by forming hydrogen bonds and inhibited the activity of same Conclusions: MBL production is one of the major causes of enzyme. IRPA. Increasing resistance to polymyxin B is grave. Due to ac- Conclusions: The present in-vivo and in-silico studies proved quisition of MBL strains, MDR P. aeruginosa has become endemic that compounds present in fruits of W. coagulans are predicted as in tertiary setups. potent inhibitors of HMGCR that could be used for the develop- ANTIBIOTIC RESISTANCE IN NEONATAL SEPSIS – STUDY ON ment of drug for hypercholesterolemia. LOW BIRTH CASES FROM KARACHI-PAKISTAN CARDIOPROTECTIVE EFFECT OF MODIFIED BONE MARROW Mirza Tasawer Baig1*, Ali Akbar Sial1, Zahida Memon2and Nusrat MESENCHYMAL STEM CELLS AFTER TRANSPLANTATION IN RAT Bano3 MODEL OF MYOCARDIAL INFARCTION: AN APPLIED APPROACH 1Department of Pharmacy Practice, Faculty of Pharmacy, Ziauddin TOWARDS CELLULAR THERAPY University Karachi,2Department of Pharmacology, Faculty of Phar- Nadia Naeem1, Muhammad Aleem Akhter1, Irfan Khan1, Hana’a macy, Ziauddin University Karachi,3Department of Pharmacology, Iqbal1, Kanwal Haneef1, Nurul Kabir1and Asmat Salim1 Faculty of Medicine, Ziauddin University, Karachi, Pakistan. 1Dr. Panjwani Center for Molecular Medicine Drug Research, In- Introduction and Objectives: Premature neonates are often ternational Center for Chemical and Biological Sciences, University of immunocompromised and readily susceptible to infectious dis- Karachi, Karachi, Pakistan. eases. They are deficient in antibodies whereas neonatal neu- Introduction and Objectives: Myocardial infarction is in- trophils are immature and depleted (2 fold in circulatory pool). creasing cause of death worldwide. In South Asia, cardiac diseases Neonatal anatomic barriers are susceptible and hence infection are increasing at a rate greater than in other part of the world. control poses significant challenge. The objective of this study is to Regenerative medicine involves the repair or regeneration of an assess the common pathogens isolated in neonatal sepsis and organ, tissue or cells in order to restore an impaired function of the specify the antibiotic resistance pattern. tissue. The cellular therapy offers a promising therapeutic strategy Methods: The study was conducted in the neonatal ICU of two for the treatment of different diseases including myocardial in- public sector tertiary care hospitals inclusive of 234 cases. Pre- farction. Mesenchymal stem cells (MSCs) are having immense mature neonates with clinically confirmed bacterial infection were importance in the emerging field of regenerative medicine and included comprising of 48.6% cases of Early onset neonatal sepsis have tremendous capability for cellular therapy and tissue en- (EONS) with obstetrical complication and fulminant, multisystem gineering. MSCs can be converted into cardiomyocytes by treating N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 203 them with DNA demethylating agents. Our aim is to investigate the when HslV dodecamer is flanked at both sides by HslU hexamers cardioprotective effect of pretreated MSCs when transplanted in and the carboxy termini of HslU subunits intercalate into a clefts infarcted myocardium of rat. between two adjacent HslV subunits. This intercalation is followed Methods: In the present study, rat mesenchymal stem cells by conformational change in HslV active site resulting in its al- were treated with DNA-demethylating agents. The optimized losteric activation. We for the first time report certain small mo- concentrations of these compounds were added separately into lecules which are capable of activating HslV peptidase in the ab- the culture medium and the treated cells were analyzed for the sence of its natural activator, HslU. expression of cardiac specific genes by RT-PCR and cardiac specific The benzimidazole, quinazoline and chromone derivatives proteins by immunocyto-chemistry and flow cytometry. The were suggested by ligand docking studies to bind at the carboxy treated MSCs were then transplanted to the rat model of myo- terminus binding pocket of HslV in a manner similar to HslU cardial infarction and cardiac functional studies were performed carboxy terminus. This was confirmed by HslV activation assays by echocardiography. with these compounds that gave ED50 in micromolar range (0.6– Results: We have found expressions of cardiac specific proteins 1.5 μM). and genes in the modified mesenchymal cells both at genetic and Results: The results showed that small, extracellular non- protein levels. Transplantation of modified MSCs into infarcted peptidic molecules can allosterically activate the peptide hydro- myocardium of rat improved the cardiac left ventricular function lytic activity of HslV, which in turn would initiate intracellular possibly by enhancing the rate of bone marrow stem cell differ- proteolysis. entiation into mature cardiomyocytes in the infarcted heart. SENSITIVITY PATTERN OF MORAXELLA CATARRHALIS ISO- Conclusions: We therefore, conclude that pre-treating auto- LATED FROM HOSPITALIZED PATIENTS logous mesenchymal stem cells before transplantation may in- Naheed A.Irfan1, Maha Jamil1, Aziz Fatima1and Syedda Gufrana crease the likelihood of successful regeneration of damaged Nadeem1 myocardium. The study put forth another valuable aspect that 1Jinnah University for Women, Karachi, Pakistan. would serve as a tool for modified cellular therapy. Introduction and Objectives: Moraxella catarrhalis (formerly ANTIPLATELET AGGREGATING AND MEMBRANE STABILIZING known as Branhamella catarrhalis) has emerged as a significant PROPERTIES OF ARECA NUT bacterial pathogen of humans over the past two decades. Over the Shagufta Khan1and Ahsana Dar2 same period, studies have revealed its involvement in respiratory 1Department of Biological and Biomedical Sciences, Aga Khan (e.g., sinusitis, otitis media, bronchitis, and pneumonia) and ocular University Hospital, Medical College, Karachi, Pakistan,2HEJ Research infections in children and in laryngitis, bronchitis, and pneumonia Institute of Chemistry, ICCBS, PCMD, University of Karachi, Karachi, in adults. It has discovered an increasing number of Β-lactamase- Pakistan. positive strains. We aim to find out the sensitivity pattern of Introduction and Objectives: Platelet aggregation inhibitors Moraxella catarrhalis isolated from sputum. are one of the key preventive and therapeutic agents for cardio- Methods: Total 124 sputum samples were analyzed. Con- vascular diseases. Areca nut is used in folk medicine for treatment firmation done by using chocolate agar a typical golden yellow of various ailments. Present study was conducted to investigate which can be distinguished by hockey puck like colony. Further the antiplatelet aggregating and membrane stabilizing activities of identifications were done by gram staining, oxidase test and bio- areca nut extract and its fractions using platelet aggregation and chemical test. Antibiogram was done by CLSI method. Statistical membrane stabilization assays. analysis was also performed using SPSS version 17. Methods: To determine the effect of areca extract and its Results: Sensitivity pattern showed AMC 99%, TE 49%, CRO 01%, fractions on platelet aggregation, blood was taken from healthy E 30%, SXT 04%, CIP 50%, CEF 84%. human volunteers and platelet aggregation was monitored. Conclusions: To control the resistivity of these organisms some Membrane stabilizing properties were assessed using human red preventive measurements like proper identification and selective blood cells. medication should be taken. Results: Our results showed that areca extract was effective ASSESSMENT OF HCV GENOTYPES GENE FLOW HYBRIDIZA- against arachidonic acid induced platelet aggregation in dose de- TION AND ITS COMPARSION WITH CONVENTIONAL PCR pendent manner leading to complete inhibition at 10 mg/ml dose. Nazish Jaffar1, Rubina Ghani2and Saba Sattar1 Data obtained from the present study showed that the areca ex- 1Jinnah Sindh Medical University, Karachi, Pakistan,2Pathological tract (EC50= 91 μg/ml) and its aqueous fraction (EC50= 55 μg/ml) and Molecular Laboratories, Karachi, Pakistan. possess significant membrane stabilizing activity in a dose de- Background: Nucleotide sequence analysis of hepatitis C virus pendent manner (IC50= 100 μg/ml). (HCV) strains showed substantial variability leading to a classifi- Conclusions: These findings suggest that areca nut extract, cation into several genotypes and subtypes. ethylacetate fraction and aqueous fraction possess significant an- Objective: The objectives of the present study were the tiplatelet aggregating and membrane stabilizing properties, and knowledge of the HCV genotypes in Pakistan, and determining aqueous fraction was most effective which may be of clinical value. their prevalence. However, further studies are required to identify its active com- Methods: During the course of this study, HCV-RNA positive pound(s) via bioassay guided-fractionation method. sera samples from 200 chronically infected patients were char- SMALL MOLECULAR ACTIVATORS OF PROTEASOME-RELATED acterized by genotyping assay and Automated Sequence Analysis. HSLV PEPTIDASE Genotyping assay utilized type-specific primers for amplification Yasmeen Rashid1 of the core region, where as sequencing was done for 5’Noncoding 1Department of Biochemistry, University of Karachi, Karachi, region of HCV using ABI 3100 Sequencing Analyzer. Pakistan. Results: During our study we came across that genotype 3a Introduction and Objectives: The HslVU or prokaryotic mini- was the most prevalent (36%) followed with 3b (26%), 3c (3.5%), 3d proteasome is two component system based on HslV peptidase (4%), 1a (2.5%), 1b (1%), 1c (2.5%), 1d (0.5%), 2a (12%), 2b (0.5%), 2c and HslU ATPase. Like proteasome it is involved in intracellular (0.5%), 4 (0.5%) and 4a (1.5%). Genotypes 3d, 1c, 1d and 2c were proteins degradation. Due to its occurrence in pathogenic mi- reported for the first time from Pakistan crobes and absence in human beings, it is considered as a potential Conclusion: Advances in the field of molecular biology have antimicrobial drug target. The functional HslVU complex forms provided rapid diagnostic tools that have reduced the turnaround 204 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 times for detecting HCV genotype by using “Flow-through” hy- 1Faculty of Pharmacy, Ziauddin University, Karachi, Pakistan. bridization in Pakistan. Introduction and Objectives: Over the past few years, herbal GALECTIN-3 AND WNT SIGNALLING IN MYOCARDIAL INFARCT remedies and medicinal plant entities are of great interest in HEALING: IS THERE A CONNECTION? various region of the world for their therapeutic and preventive Satwat Hashmi1 and Suhail Al-Salam2 effects in various cardiovascular ailments like atherosclerosis. 1Department of Biological and Biomedical Sciences, Aga Khan Numerous animal models were studied extensively for elucidation University, Karachi, Pakistan,2Department of Pathology, College of of these plant component effects on different organs. Citrus Medicine and Health Sciences, UAE University, UAE. Paradisi (C. paradisi ) due to its antioxidant and anti-inflammatory Introduction and Objectives: Myocardial infarction (MI) leads potential has been selected in this study to evaluate the effect of to complex repair processes including scarring and fibrosis. Ga- this compound on coagulation and anticoagulation factors. C. lectin-3 (Gal-3) is a beta-galactoside binding lectin responsible for paradisi contains high concentrations of hesperidin, naringin, li- cardiac fibrogenesis and adverse left ventricular (LV) remodeling. monene and related flavonoids. Canonical Wnt signaling in myofibroblasts following MI also Methods: Doses were selected in the range of 0.1-0.5 ml/kg. modulate wound healing and fibrosis. As Gal-3 and Wnt 3a play a Aspirin and warfarin 150mg/kg and 5mg/kg-10mg/kg respectively significant role in the post MI sequel we aim to measure their were used as standard anticoagulants. Animals were divided into concentration in murine heart at 7 week post Ischemia/reperfu- six groups with ten rabbits in each group. sion injury and to determine their correlation. Results: At median dose level, significant enhancement in Methods: Male C57B6/J mice were used for myocardial IR in- thrombin, pro-thrombin time was observed in comparison to jury by temporarily ligating the left anterior descending artery of control, while fibrinogen levels were significantly reduced with the heart for 30 minutes to create ischemia/infarction followed by warfarin. Furthermore, substantial inhibition of platelet aggrega- restoration of blood flow. Samples collected 1 week post IR injury tion by adenosine phosphate, collagen, epinephrine and arachi- point was processed for ELISA to detect Gal-3 and Wnt-3a levels in donic acid was observed at median C. paradise dose (MCPD). the heart. Conclusions: Results of this study have shown an effective Results: Gal-3 and Wnt 3a levels were significantly increased in anticoagulant activity. Thus the patients, who are at risk of car- the LV heart tissue at 1 week post IR compared to sham operated diovascular event development may additionally be advised to mice. There was a strong and significant positive correlation ob- consume grape fruit juices in their diet to reduce the chances of served between Gal-3 and Wnt-3a at 1week post IR injury. mortality and associated morbidities. Conclusions: Gal-3 and Wnt-3a levels were increased in 1 CONVENTIONAL CLINICAL AND PROGNOSTIC VARIABLES IN week post myocardial IR injury in conjunction with intrinsic car- 150 ORAL SQUAMOUS CELL CARCINOMA CASES FROM THE IN- diac repair. Understanding the mechanisms of this repair process DIGENOUS POPULATION OF KARACHI could lead to optimal management and prevention or delay in the Mohiuddin Alamgir1 onset of heart failure. 1Bahria University, Karachi, Pakistan. STD PATHOGENS DETERMINED IN SEMEN USING PCR AND Objectives: To evaluate conventional clinical and prognostic “FLOW-THROUGH” HYBRIDIZATION TECHNOLOGY variables in Oral Squamous Cell Carcinoma (OSCC) cases from the Rubina Ghani1 indigenous population of Karachi. 1Department of Biochemistry, Pathological and Molecular La- Methods: Common clinical and pathological parameters re- boratories, Department of Physiology, Jinnah Medical and Dental lated to poor prognosis of 150 OSCC cases presented at two major College, Karachi, Pakistan. treatment facilities of Karachi were evaluated. The reporting in- Background: The prevalence of sexually transmitted Disease cluded demographic details and variables like intra-oral subsites, (STDs) in health care facilities for diagnosis of common pathogens, clinical stage and histological grades. Recurrence of tumor after those causing infertility like Chlamydia trachomatis, Neisseria initial resection was also documented. gonorrhoeae and Mycoplasma hominis. Genital wart is a highly Results: The patient’s population comprised of 98 males and 52 contagious sexually transmitted disease caused by some sub-types females. The mean age was 47.1 + 12.22 (range: 20-78 years). of human papillomavirus (HPV). Maximum numbers were seen in the 41–50 years age group. Urdu- Methods: Semen samples were obtained by masturbation into speaking community was the most affected ethnic group (n=75). sterile containers after sexual abstinence of 48 to 72 hours. Sam- Histopathological analysis revealed that majority of cases was ples were subjected to semen analysis within one hour of collec- moderately differentiated tumors (59%) with clinical stage II (35%) tion. The concentrations of sperm as well as sperm motility were or IV (29%). The most common intra-oral subsite came out to be also determined. DNA extraction was extracted of all the samples buccal mucosa of cheeks (56%) followed by lateral borders of and the PCR assay was performed. The amplicons are subsequently tongue (21%), lips (13%), alveolar (6%), palate (2.6%) floor of mouth hybridized to pathogen-specific capturing probes via “Flow- (1.3%), etc. Recurrence was observed in 8 out of 150 cases. All through” hybridization. patients underwent primary resection ± neck dissection and re- Result: During our study we came across with the STI patho- construction where possible. gens present in our population and the reason for infertility was Conclusions: Overall experience with oral squamous cell car- the main cause. When Chlamydia trachomatis and Neisseria go- cinoma shows that it has a high tendency for local invasion as well norrhoeae were detected in their wife’s were also screened and as dissemination to regional lymph nodes, i.e. cervical lymph these STI pathogens were identified. nodes, both are associated with a poor prognosis. Preventable risk Conclusions: The main route for the transfer of STI pathogens factor of tobacco chewing has been observed in majority of these were the men special those who visited commercial sex workers cases. as they were working in other cities and the complained for in- Poster Presentations fertility. Screening for bacterial STI pathogens, Mycoplasma ho- ROLE OF MUSCARINIC G-PROTEIN COUPLED RECEPTOR minis, Chlamydia trachomatis and Neisseria gonorrhoeae are (GPCR) IN PROSTATE CANCER GROWTH AND SURVIVAL strongly recommended because these pathogens can cause serious Abdul Mannan Baig1and Farhat Abbas2 reproductive complications. 1Department of Biological and Biomedical Sciences,1Department EFFECT OF CITRUS PARADISI ON COAGULATION MECHANISM of Pathology and Microbiology,1Department of Surgery,1Aga Khan Neelam Mallick1, Ali Akbar Sial1and Huma Ali1 University, Karachi, Pakistan. N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 205

Introduction: Despite advances in early detection and treat- therapeutic options. We aimed to investigate the alterations in ment, prostate cancer remains one of the most frequently diag- protein expression and association of CDK10 and ETS2 during nosed cancers and a leading cause of cancer death in males corneal epithelial migration and to demonstrate the networks of worldwide. Much work has been done on androgen receptors in the total identified proteins with potential dual functions. relation to proliferation and survival, and it has been shown that Methods: In this study, human corneal epithelial cells lines cancers of this organ usually turn androgen insensitive and re- (HCEC) have been used for wound healing model. Mechanical sistant to therapy. There has been very little, if any, work done on wound was made in HCEC lines and healing was monitored at 24, the precise contribution of cell surface muscarinic GPCRs in 48 and 72 hours of post wounding. Epithelium was scrapped at 24, prostate cancer growth. 48 and 72 hours, followed by protein quantification using BCA kit. Methods: Here we determined the role of the M1 receptor in The wounded and unwounded cells were subjected to SDS-PAGE prostate cancer cell proliferation. The treatment of PC-3 cells with and two-dimensional electrophoresis (2DE). Mass Spectrometry selective M1 receptor antagonists, Dicyclomine exhibited anti- (MALDI TOF) was done to identify the proteins through protein proliferative effects using growth assays, while M1 receptor ago- database searches. The identified protein were further analyzed nist, Pilocarpine exhibited increased proliferation compared with and validated by western blot analysis. A further insight into the the untreated cells. links among the identified proteins and their functional roles were Results and Conclusions: Further studies will dissect mole- analyzed by STRING 8.3 software. cular pathways to determine the role of the M1 receptor in an- Results: A significant finding of the present study is the iden- drogen sensitive and androgen insensitive prostate cancer cell tification of Cdk10, EFNB3, RAB 34, RRAS, HSP2 and HSP90 in growth and proliferation. The results of this work may help inform healing corneal epithelium at active phase of migration. Expres- clinical and public health interventions for prostate cancer by sion of CDK10 and ETS2 were validated using antibody by western providing insights into the biology of prostate cancer progression. blot. Interaction association network analysis further confirms the CO AND CONTRA-REGULATION OF GLYCOSYLATED PROTEINS close interacting relationship between CDK10 and ETS2 proteins. IN THREE DISTINCT REGIONS OF SCHIZOPHRENICS BRAIN Conclusions: These findings are one step forward in identifying Ayesha Khan1, Nikhat Ahmed2, Beena Hasan1and Shamim the mechanism of wound repair or re-epithelialization. Associa- Mushtaq2 tion of CDK10 and ETS2 in migrating samples suggested that they 1Neurochemistry Research Laboratory, Department of Biochem- have positively regulated the cell cycle and cell proliferation and istry, University of Karachi, Karachi, Pakistan,2Department of Re- hence mediate the wound healing. This study may also increase search / Biochemistry, Ziauddin University, Karachi, Pakistan. the understanding of normal and abnormal corneal function with Introduction and Objectives: Schizophrenia, a multifactorial likely relevance to corneal disease and transplants. disorder exhibits diverse neuropathological aberrations with al- FUNCTIONAL ASSOCIATION OF BAD AND 14-3-3 PROTEINS IN tered protein expression and post translational modifications as a SCHIZOPHRENIC PATIENTS surfacing evidence that may also contribute to its pathophysiology. Sobia Manzoor1, Shamim Mushtaq2, Ayesha Khan1and Nikhat To investigate the integrated picture of molecular changes and Ahmed2 potential alterations in expression of specific glycosylated proteins 1Department of Biochemistry, University of Karachi, in different brain regions compared to physiologically normal Pakistan,2Department of Research, Ziauddin Medical University, brains. We aimed to provide more holistic view of three brain Karachi, Pakistan. regions; and their dynamic cross-talk providing insights into the Introduction and Objectives: Schizophrenia is a multifactorial underlying molecular mechanisms associated with schizophrenia. disease which involves the exogenous and endogenous interaction Methods: Present study entails, differential proteomic analysis and is characterized by neuronal reduction, dendrite deficits, de- of autopsied brain regions of schizophrenic; substantia nigra, creased synaptic markers and alteration in synaptic circuitry. cortex, and hippocampus (n=7 each), by using sodium dodecyl Schizophrenia is a neurodevelopmental disorder but apoptosis sulphate polyacrylamide gel electrophoresis coupled with im- may contribute to the pathophysiology of schizophrenia due to munoblot and DIG (digoxigenin) labelling followed by ESI-QTOF progressive clinical deterioration and subtle neurostructural MS analysis for validation. changes following the onset of psychosis. Apoptosis is a mechan- Results: We have identified fourteen glycosylated protein ism of cell death, is regulated by a complex cascade of pro and components with altered expression among the brain regions. The anti-apoptotic proteins. Dysregulation of apoptosis in several cor- 50KDa (Glial fibrillary acidic protein, GFAP) and 84KDa (mi- tical regions of schizophrenia indicates that apoptotic vulnerability tochondrial inner membrane protein) are contra-regulated be- is increased. Pro-apoptotic protein BAD (Bcl-2 associated death tween substantia nigra and cortex. While T-complex protein 1 promoter) interacts with anti-apoptotic BCL-2 and inhibit its ac- subunit zeta of 58KDa is co-regulated between substantia nigra tivity while 14-3-3s binds with BAD and inhibit apoptosis that it and hippocampus. The remaining five proteins identified with act as an ant apoptotic protein. Various proteomic based studies significantly altered glycosylation intensity in the specific brain shown reduction of 14-3-3 proteins in schizophrenic brains. In this regions give evidence of their explicit regional function. Ad- study we aimed to identify and validate the differential expression ditionally, nine proteins commonly appeared non-glycosylated in of BAD and 14-3-3s by western blotting and their association the three studied brain regions. network with other proteins by String 10, Qiagen pathway data EXPRESSION AND ASSOCIATION OF CDK10 WITH ETS2 DUR- bases. ING HUMAN CORNEAL WOUND HEALING Methods: Expression of BAD and 14-3-3 proteins from autop- Meraj Zehra1, Shamim Mushtaq2and Nikhat Ahmed Siddiqui2 sied schizophrenic brain cortex were validated by western blotting 1Department of Biochemistry, Karachi University, and In Silico analysis was done to see the association with other Pakistan,2Department of Research/Biochemistry, Ziauddin University, proteins by using String database, Qiagen pathway. Karachi, Pakistan. Results: We have identified differential expression of BAD and Introduction and Objectives: Corneal related complications 14-3-3 proteins in cortex region of schizophrenic human brain by are major health concerns worldwide because its progression is western blotting. Interaction association network between BAD associated with significant impaired vision. Therefore, there is an and 14-3-3 (YWHAQ,YWHAH,YWHAE,YWHAB,YWHAZ,Y-WHAG) urgent need to develop reliable understanding of the underlying was analyzed by string software. Qiagen pathway further confirms mechanism of corneal epithelial wound healing to apply the close interacting relationship between BAD and 14-3-3 206 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 proteins in apoptotic pathway related to schizophrenic patients. compared to 0.15 μg/ml in comparison group. Conclusions: Apoptotic proteins BAD and 14-3-3s association Conclusions: The increase in nephrin levels from 0.86μg/ml in in Schizophrenic brain cortex region provide insight into the patients with normal albuminuria to 47.6μg/ml in patients with apoptotic pathways which play role in the pathophysiology of macroalbuminuria suggests that Nephrin precedes albumin in schizophrenia which could lead to development of therapeutic urine predicting early signs of kidney damage. drugs to target apoptotic proteins and their interacting partners. BIOINFORMATICS NAVIGATION PREDICTED STRUCTURE- KISSPEPTIN AND UNEXPLAINED INFERTILITY FUNCTION DYNAMICS OF CALPAIN10 ISOFORMS Aaida Mumtaz1, Aqsa Khalid1, Zahir Hussain2, Faiza Alam1and Farina Hanif1, Bushra Chaudhry1and Kausar Saboohi1 Rehana Rehman1 1Department of Biological and Biomedical Sciences, The Aga Khan 1Aga Khan University, Karachi, Pakistan,2Umm Al-Qura University, University Medical College, Karachi, Pakistan. Kingdom of Saudi Arabia. Introduction and Objectives: Calpain10, a member of cysteine Introduction and Objectives: To compare Kisspeptin (KP) le- proteinase family, is present in eight different isoforms (a-h) exact vels in females with infertility to male, female and unexplained mechanism through which it influences the insulin secretion and causes and associate it with success of treatment after ICSI. action is scanty. Methods: Cross sectional study carried from August 2014 till Methods: We have applied bioinformatics approach to en- May 2015 included all categories of infertility with duration 4 visage its mechanism of action and possible involvement in dis- two years, female age between 20 to 50 years and body mass eases. National Center for Biotechnology Information Blast was index 418 o35 kg/m2. Down-regulation of ovaries was followed used for humans’ nucleotide and protein sequence alignments of by calculated stimulation, ovulation induction, oocytes retrieval, the calpain10 isoforms a, c and g. Further, NCBI conserved domain ICSI, in vitro maturation of embryos and finally blastocysts trans- tool and CDART were used for other species. fer. KP levels were measured by enzyme linked immuno sorbent Results: Results of blast showed 100% nucleotide whereas 91% assay on day of ovulation induction. Failure of procedure was de- and 97% amino acid sequence identity among isoforms of Homo tected by beta human chorionic gonadotropin o5-25mIU/ml sapiens’ calpain10 c and g with calpain10 a. There was 82% query (non-pregnant) whereas females with levels 425mIU/ml and no coverage of calpain10 c and 17% of calpain10 g with calpain10 a. cardiac activity had preclinical abortions. Clinical pregnancy group Conserved domain analyses of peptide sequences indicated do- had confirmed cardiac activity on trans vaginal scan. Data was mains IIa and IIb have significant homology with the catalytic analysed using SPSS 15. domain of the cysteine protease superfamily in all three isoforms. Results: Females who completed the procedure (168) were The domain III and IV of isoform a and domain III of isoform c were stratified; 40 (24%), 66 (39%) and 62 (37%) with male, female and found homologous with linker C2 like subdomain III of mu- unexplained infertility. The KP levels were 285.38 ± 23.96 ng/L calpain. (mean ±SD). The females with unexplained infertility had lowest Conclusion: It is predicted that calpain10 a and c isoforms may KP (101.4 ± 11.96) which was statistically significant (po0.05). The have a role in developing diseases such as neurodegenerative, results declared 66 (39%) females as non-pregnant, 30 (18%) with cardio-vascular, cataract, osteopenia and cancer. Whereas isoform preclinical abortions and 72 (43%) with clinical pregnancies. Out of g may exhibit protease activities having papa-in-like domain only. non-pregnant females 42 (64%) had unexplained cause of TP53 PROTEIN OVEREXPRESSION IN ORAL SQUAMOUS CELL infertility. CARCINOMA (OSCC): CORRELATION WITH HISTOLOGIC VARI- Conclusions: Low KP levels in unexplained females explains ABLES AND 5 YEARS SURVIVAL IN PAKISTANI PATIENTS the peripheral role of the neuropeptide, deficiency of e=which Adnan Ali1, Sohail Awan1, Omar Irfan2and Hamza Khan2 leads to failure of implantation after ICSI. 1Aga Khan University Hospital, Karachi, Pakistan,2Aga Khan Uni- RELIABILITY OF NEPHRIN AS AN EARLY DIAGNOSTIC BIO- versity Medical College Karachi, Pakistan. MARKER FOR SCREENING DIABETIC NEPHROPATHY Introduction and Objectives: Alteration of TP53 gene in oral Moazzam A Shahid1, Saeeda Baig1, Adnan M Zubairi1and Farha squamous cell carcinoma (OSCC) is believed to be associated with Ahmed1 reduced overall survival (OS), the 5 year prognosis in our study 1Ziauddin University, Karachi, Pakistan. and the disease-free survival (DFS). The aim of this study is to Introduction and Objectives: Diabetic nephropathy occurs as a determine whether TP53 protein over expression in OSCC is a result of proximal tubule dysfunction with podocyte damage due prognostic indicator of survival in Pakistani cancer patients along to increased advanced glycation end-products insults in diabetes. with its correlation with risk factors including smoking, chewing Nephrin, one of the three proteins that make up the podocyte habits, histological variables like grade and stage of the tumor in a architecture, is excreted foremost following renal damage. The aim high risk population. of this study was to evaluate the efficiency and reliability of ne- Methods: A total of 140 patients of OSCC were part of our phrin for an early biomarker of kidney damage in diabetic patients. study. TP53 protein over expression was investigated by means of Methods: Urine samples (78) were collected from diabetic immunohistochemistry. center. Protein and glucose were determined by Dipstick. The Results Overexpression of p53 protein was observed in 75 patients were grouped on the basis of Albumin/creatinine ratio patients (54%) using a threshold of 10% stained tumor nuclei. Pa- (UACR) as normoalbuminuric, microalbuminuric and macro- tients with p53 negative tumors had improved OS when compared albuminuric with UACR less than 30 mg/g, from 30-300mg/g, and with patients with p53 positive tumors. This difference was sta- above 300mg/g respectively. ELISA (Exocell USA) was used for tistically significant (p=0.036) in univariate Cox regression analysis Nephrin estimation. Statistical evaluation was done. however, it lost its worth in the multivariate analysis. Results: Nephrinuria was present in 70 (89.7%) out of 78 dia- Conclusion: This works supports that patients with p53 over- betic patients including 35 (81.4%) normoalbuminurics, 5 (6.4%) expression had a significantly poor overall survival compared to micro-albuminurics and all 5 of macroalbuminurics (po0.027). p53 negative patients. However, p53 overexpression was not as- When associated with duration of diabetes, 21 (91.3%) positive sociated with patient’s disease free survival. with less than three years of diabetes, 24 (92.3%) with three to CARDIOVASCULAR EFFECTS OF AQUEOUS-METHANOLIC EX- seven years and 25 (86.2%) in more than seven years (po0.039). TRACT OF BERGENIA LIGULATA IN EXPERIMENTAL ANIMALS Nephrin levels were found increasing from Normo-(0.86ug/ml) to Hasan Salman Siddiqi1and Mohammad Ahsan Sohail1 Micro-(11.6μg/ml) to Macroalbuminuria group (47.6μg/ml), 1Department of Biological and Biomedical Sciences, Medical N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 207

College, Aga Khan University, Karachi, Pakistan. hepatitis C patients is found to be associated with novel Hepatitis Introduction and Objectives: Prevalence of hypertension, a G virus. major risk factor for diseases like ischemic heart disease and cer- COMBINATION THERAPY OF NON-STEROIDAL ANTI-IN- ebrovascular stroke, is on the rise. The aim of this study was to FLAMMATORY DRUGS AND ANTI-OXIDANTS: AN EFFECTIVE RE- explore the cardiovascular effects of the aqueous-methanolic ex- GIMEN FOR INFLAMMATORY DISORDERS? tract of Bergenia Ligulata rhizome (Bl.Cr), to provide pharmaco- Shagufta Khan1, Osama Anis Silat2and Ahsana Dar3 logical basis for its medicinal use in hypertension. 1Department of Biological and Biomedical Sciences, Aga Khan Methods: Aortae from Sprague-Dawley rats and atria from University Hospital, Medical College, Karachi, Pakistan,2Ziauddin guinea-pigs were placed in 5 ml and 15 ml isolated tissue bath University, Faculty of Medicine, Karachi, Pakistan,3HEJ Research In- assemblies respectively, which were maintained at 37oC with stitute of Chemistry, ICCBS, PCMD, University of Karachi, Pakistan. constant carbogen supply, filled with physiological salt solution Introduction and Objectives: Non-steroidal anti-inflammatory (Kreb’s solution) and connected to a force transducer and Power- drugs are of therapeutic value for inflammatory disorders. Reactive Lab attached with a computer. Oxygen Species may initiate and/or aggravate inflammation. In- Results: Bl.Cr, in isolated guinea-pig atria, equally inhibited formation regarding the interaction of vitamin E with NSAIDs in force and rate of spontaneous atrial contractions. When tested on inflammation is not well documented and is controversial. phenylephrine (PE, 1 μM) and K+ (80 mM)-induced vasoconstric- Methods: This study was conducted to determine the effect of tion, Bl.Cr caused a concentration-dependent relaxation and also aspirin with vitamin E supplementation in inflammation, and to caused a suppression of PE (1 μM) control peaks in Ca2+-free explore the possible interactions of vitamin E with one drug from medium. each sub group of NSAIDs with respect to their anti-inflammatory Conclusions: These data indicate that Bl.Cr exhibits cardio- activity using animal model of inflammation, DNA degradation suppressant and vaso-dilatory properties. The vasodilator effect of and lipid peroxidation assays. the plant extract is mediated through inhibition of Ca++ influx via Results: Our results showed that vitamin E caused 10% in- membranous Ca++ channels as well as Ca++ release from in- hibition in edema at 400 mg/kg dose while aspirin elicited com- tracellular stores. Further studies are required to elaborate the plete inhibition in edema at 200 mg/kg dose. Co-administration of anti-hypertensive activity of the plant. different doses of aspirin with vitamin E showed a dose-depen- ASSOCIATION OF HEPATITIS G WITH LIVER DYSFUNCTION IN dent increase in edema inhibition. At 100 mg/kg aspirin showed TREATMENT RESPONDERS HEPATITIS C PATIENTS ~50% inhibition in edema. While the same dose of aspirin with Uzma Najam1, Syed Tousif Ahmed1and Rubina Ghani2 vitamin E supplementation (200 mg/kg) showed a significant in- 1Department of Physiology, Ziauddin University, Karachi, crease (~25%) in its anti-inflammatory effect. Similar results were Pakistan,2Department of Biochemistry, Baqai Medical University, obtained with DNA degradation and lipid peroxidation assays. Karachi, Pakistan. Conclusions: In conclusion, aspirin, diclofenac sodium, and Background: About 10 million Pakistani population is infected celecoxib, with vitamin E supplementation, showed significant with Hepatitis C virus (HCV). The prevalence is even more pro- synergistic effects that could be used as an effective preventive nounced among high risk population. Combination of standard and therapeutic regimen for inflammatory diseases. interferon and ribavirin is still the first line therapy with sustained CLINICAL EFFICIENCY AND COST EFFECTIVENESS OF MAC- viral response (SVR) up to 40-50%. Pool of relapsers and non-re- ROPROLACTIN SCREENING IN HYPERPROLACTINEMIC PATIENTS sponders is increasing in Pakistan and needs treatment with pe- N Sherazi1, M Z Baig2, A H Khan1, I Siddiqui1 and F Ghani1 gylated interferon plus ribavirin (peg IFN/RIB). The hepatitis G 1Section of Clinical Chemistry, Department of Pathology and virus (HGV), a recently identified member of the Flaviviridae fa- Laboratory Medicine, 2Medical College, Aga Khan University Hos- mily, can cause chronic infection in man pital, Karachi, Pakistan. Methods: 250 treatment responders Hepatitis C patients (un- Introduction and Objectives: Macroprolactin (MaPRL) is a detectable HCV RNA in the serum after 24 weeks of post treatment biologically inactive compound which leads to falsely elevated follow up) were recruited from Jinnah Post Medical College and prolactin levels. It is recommended that all sera with increased Ziauddin Hospital. total prolactin concentrations be sub fractionated by PEG pre- Liver function tests of all patients were assessed by automatic cipitation to measure the bioactive monomeric prolactin con- analyzer kits. HGV RNA was investigated in serum samples by centration to prevent misdiagnosis and unnecessary investiga- reverse transcription and polymerase chain reaction amplification tions. We aim to determine clinical efficiency and cost effective- of the 5' non-coding region of HCV and hybridisation to a specific ness of PEG screening of hyperprolactinemic sera. probe. The liver function tests of HGV RNA sero-positive and sero- Methods: In this Retrospective cross sectional study patients negative patients were compared. with high total prolactin levels were screened by PEG precipita- Results: A total of 250 treatment responder hepatitis C patients tion. Relevant diagnosis of Macroprolactinemia and True Hyper- were evaluated with a mean age 44± 5.55. Males constituted 38.4 prolactinemia was made based on their absolute PEG treated %(N=96) while female constituted 61.6% (N=154) of our study monomeric prolactin level. They were then contacted on phone group. Mean total billirubin, direct billirubin and indirect billirubin and a detailed history of their clinical symptoms along with their was 0.772 ± 0.19 mg/dl, 0.308 ± 0.99 mg/dl and 0.496 ± 0.55 mg/dl radiological workups was inquired. respectively. Mean ALT was 81.97 ± 38.03 units/L and total protein Results: Frequency of macroprolactin was 60.7% seen in 145 was g/dl 5.25 ± 0.44. Statically significant difference of mean LFTs patients and true hyperprolactinemia was observed in 94 (39.3%) was found between hepatitis G positive and negative patients with patients. More asymptomatic patients were reported in the mac- p-value = 0.001. To find out association of LFTs with hepatitis G roprolactin as compared to true hyperprolactinemic group (p = positive patients, chi-square was used. Total billirubin, direct bil- o0.05). 37 (39.4%) patients with true hyperprolactinemia had lirubin, indirect billirubin, AST, GGT, and APTT are found to be further radiological investigations done where-as only 8 (5.5%) of associated with hepatitis G with p-value of 0.001. By Log Regres- the patients with macroprolactin had undergone further radi- sion we found significant and positive association of Hepatitis G ological workups. The total use of resources in the true hyper- with Total billirubin, direct billirubin, indirect billirubin, GGT and prolactinemic group was significantly higher, 943000 PKR (10031 APTT (p-value = 0.001 at 95% CI). PKR per person) vs 418450 PKR (2884 PKR per person) (p= o Conclusions: Liver dysfunction in treatment responder 0.05). 208 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

RETINAL NERVE FIBER LAYER THICKNESS IN A SUBSET OF found to be protective against metabolic syndrome. KARACHI (PAKISTAN) POPULATION Conclusion: Prevalence of metabolic syndrome is increasing in Sahrish Mukhtar1, Nuzhat Hassan1, Zafar Dawood1and Nosheen our adolescent population which demands screening from child- Zehra1 hood and early adolescence. Early screening, identification and 1Ziauddin University, Karachi, Pakistan. lifestyle interventions will decrease the morbidity from chronic Introduction and Objectives: To provide the normal range of illnesses like Coronary artery disease and Diabetes mellitus. retinal nerve fiber layer (RNFL) thickness in a subset of Karachi ASSOCIATION OF VITAMIN D WITH OUTCOME AFTER INTRA population by Spectralis OCT and to evaluate the effects of age and CYTOPLASMIC SPERM INJECTION gender on it. Umme Hani Abdullah1, Salima Lalani1, Sara Arif2, Fatima Methods: 300 eyes from 150 healthy subjects aged 40 years Syed3and Rehana Rehman3 and above with no ocular pathologies were examined using 1Aga Khan University, Karachi, Pakistan,2Dow University of Health standard protocols by a single examiner. Subjects with history of Sciences, Karachi, Pakistan,3Jinnah Sindh Medical University, Karachi, diabetic or hypertensive retinopathy, raised intraocular pressure Pakistan. (4 21mmHg) and previous intraocular or laser surgery were ex- Introduction and Objectives: To observe effects of vitamin D cluded from the study. The mean retinal nerve fiber layer thickness levels on pregnancy outcome after intra cytoplasmic sperm in- was calculated and was correlated with age and gender difference. jection (ICSI). Results: The mean global retinal nerve fiber layer thickness was Methods: It was a cross sectional study conducted in Australian found to be 99.02 ± 9.08 μm in our set of population. Out of four Concept Infertility Medical Centre from July 2011 to August 2014. quadrants the maximum RNFL thickness was found in inferior Estimation of 25 hydroxy cholecalciferol (25-OHD) of consented quadrant (126.45 ± 16.23 μm) followed by the thickness of 121.50 females (252) was done before treatment protocol for ICSI. Results ± 15.03 μm in superior quadrant, 70 ± 14 μm in nasal quadrant and of ß hCG performed 14 days after embryo transfer categorized 68.90 ± 13.10 μm in temporal quadrant. We found strong negative groups; Pregnant with ß hCG more than 25 IU/mL and rest in- correlation of RNFL thickness with age (P= o 0.001) and not cluded in non-pregnant group. Both groups were compared by significant relation with gender (P= 0.8). independent sample t-test and Pearson Chi Square test. Binary Conclusions: Keeping in mind the variations in RNFL thickness Logistic Regression Analysis was used to estimate odds ratio of with ethnic differences, this study provides the normal values of pregnancy outcome with its predictors. RNFL thickness according to our set of population. It is concluded Results: The mean value of 25-OHD, number of oocytes, ferti- that RNFL thickness decreases significantly with increasing age but lized oocytes and endometrial thickness was significantly higher gender had no significant effect on it. in pregnant women. A significant positive association of 25-OHD PREVALENCE OF METABOLIC SYNDROME IN ADOLESCENTS with clinical pregnancy and thickness of endometrium was ob- /FIRST YEAR MEDICAL STUDENTS OF A PUBLIC SECTOR MEDICAL served. After adjustment with female age and BMI positive asso- COLLEGE IN QUETTA, PAKISTAN ciation of vitamin D with endometrial thickness was observed. Faisal Shahzad1, Aiesha Ishaque2and Noman ul Haq3 Conclusion: Deficiency of (25-OH) in females hinders the ac- 1Bolan Medical College, Quetta, Pakistan,2Family Medicine, complishment of optimal endometrial thickness required for im- Ziauddin University, Karachi, Pakistan,3Pharmacy Practice, University plantation of embryo after ICSI. The improvement in vitamin D of Balochistan, Quetta, Pakistan. status can thus improve success results in assisted reproductive Introduction and Objectives: Metabolic syndrome is a dis- clinics. order of energy utilization and storage. According to the Interna- COMPARISON CLINICAL OUTCOME AND COST-EFFECTIVENESS tional Diabetes Federation, metabolic syndrome in adolescents OF TEGAFUR AND CAPECITABINE FOR THE TREATMENT OF ME- more than 16 years is defined as “the presence in an individual of TASTATIC COLORECTAL CANCER central obesity plus any two of the following four risk factors: Khola Noreen1, Imran Shaikh1and Nadia Khalid1 hypertriglyceridemia, hypertension, low high density lipoprotein 1Bahria University Medical and Dental College, Karachi, Pakistan. and high fasting glucose levels. Introduction and Objectives: The development of numerous Reports suggest that around 25% of the world’s adult popula- oral chemotherapy agents has led to a new paradigm in manage- tion suffers from metabolic syndrome. People with metabolic ment of cancer. Treatment modalities with cost effective approach syndrome are twice as likely to die and have three times higher and better clinical outcomes can be proved to be an effective chances of heart attack or stroke. strategy to reduce disease burden. The study is designed to determine the prevalence of metabolic To evaluate the clinical and cost effectiveness of oral che- syndrome in first year medical students between 17-19 years, of a motherapeutic agents Capecitabine and Tegafer as first-line public sector medical college in Quetta. treatments for patients with metastatic colorectal cancer. Methods: A Cross-sectional study was conducted on all the Methods: The analysis involved 121 patients of metastatic co- healthy first year medical students, ages from 17-19 years studying lonic carcinoma admitted at Almehrab Tibi Amdad Hospice care, in a public sector medical college in Quetta, Pakistan. Demographic Karachi between September 2013 to March 2015. The patients data were assessed by using self-administered questionnaire. selected to enrolled in present study were aged ≥18 years, both Height, weight and waist circumference were measured along sex and histologically diagnosed cases of colorectal cancer (stage 3 with fasting blood sugar, high density lipoprotein and triglycerides with primary surgical resection. While those with history of hy- by using Automated Biochemistry Analyzer. Analysis was done on persensitivity to Tegafur, fluoropyrimidines, capecitabine, or any SPSS 20. Descriptive statistics was used to present the data and other ingredients of this product, Inadequate hematopoietic logistic regression was applied to see the relationship between the function WBC 4,000/mm3; ANC 2,000/mm3; Platelet 100,000/ study variables (Po0.05). mm3, Inadequate organ function, CNS metastasis, life expectancy Results: A total of 225 were included in the study, out of which less than 3 months and those who were not willing to participate (n=127, 56.4%) were females. Mean age was 18.79 +0.38 years. were excluded. All subjects fulfilling the eligibility criteria were Prevalence of metabolic syndrome was found to be 14.2% in our randomly assigned to two groups, one group received oral cape- study sample. Increased waist circumference, fasting blood sugar, citabine (1250 mg/m2 twice daily for 14 days followed by one triglycerides were associated with an increased likelihood of ex- week gap) and other received Capsule Tegofer 500 mg daily for 28 hibiting metabolic syndrome. Male gender and high HDL were days followed by one week gap. All the patients were followed and N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 209 response was observed after 3 months, 6 months and at the end of phosphates (ALP), total bilirubin, total protein and albumin. Beside year. these, percent inhibition of LPO was also increased. Results: Patients receiving Tegafer showed significantly less Conclusion: The ESEt of C.anthelminticum was found as a he- incidence of alopecia, diarrhea and stomatitis (P o0.05) while patoprotective and antioxidant agent that could be used for the incidence of hand-foot syndrome and grade 3/4 hyperbilir- preparation of medicine in future. ubinemia were found to be significantly more as compared to SPECTRUM OF PRENEOPLASTIC AND NEOPLASTIC LESIONS capecitabine (P o0.0001). Cost of treatment per patient and per OF INTESTINE IN A TERTIARY CARE HOSPITAL OF KARACHI, cycle using oral Tegafer was less than that using capecitabine. PAKISTAN Conclusion: Tegafer showed overall improved response rates, Asma Shabbir1, Talat Mirza1, Asif Qureshi1and Abdullah Khalid1 cost effectiveness and has lesser side effects in comparison with 1Dow Institute of Medical College, Dow University of Health Sci- Capecitabine. ences, Karachi, Pakistan. RADIOLOGICAL STUDY OF THE MEAN AGE OF FUSION OF Introduction: Gastrointestinal malignancies are the global MEDIAL END OF THE CLAVICLE AS A PARAMETER OF AGE oncological problems. It is therefore imperative to investigate the Qudsia Hassan1, M Umer Memon1, Inayat-ur-RehmanKhalil1, spectrum of this pathology in Pakistan. Tehreem Ashraf1, Muhammad Mustafa Arif1and Ali Shan Iqbal1 Objective: To generate the spectrum of premalignant and ma- 1Forensic Medicine, Ziauddin University, Karachi, Pakistan. lignant lesions of small and large intestine received at Dow Di- Introduction and Objectives: Estimation of the age of a person agnostic Research and Reference Laboratory. is a problem that a forensic expert confronts in both living and Methods: The study was conducted during 2009 to 2014. All dead subjects. 21 years of age is of immense medico-legal im- the cases of preneoplastic and neoplastic lesions of intestine re- portance, as it entitles a person to be employed as a gazetted of- ceived during the period of 6 years were reviewed. The data was ficer, getting driving license for all types of vehicles, participation subjected to statistical analysis using SPSS version 22. in various sports events and is a mandatory parameter for iden- Results: A total of 64 cases were diagnosed as premalignant tification in dead. This study gives us an authentic criterion for lesions of intestine consisting of ulcerative colitis (19/29.7%), assessing 21 years age, by using closure of ossification centers at adenomatous polyp (17/26.6%), dysplasia (14/21.9%) and adenoma medial clavicular epiphysis by applying radiological method for (14/21.9%). subjects living in Karachi. About 478 cases were diagnosed as malignant lesions of in- Methods: The main objective of this study is to determine the testine as: mean age of fusion of medial end of clavicle by radiological method, to get a perfect estimator of 21 years of age. i. Adenocarcinoma grade I (65/13.6%), grade II (283/52.2%), grade Results: The mean age of fusion of medial end of the clavicle III (95/19.9%) was found to be 21±1.43 years. Significant difference was observed ii. Squamous cell carcinoma grade I (2/0.4%), grade II (7/1.5%), in male to female ratio 21.14 ± 1.41 versus 20.65 ±1.94 (P value grade III (3/0.8%) o0.05). Similarly, statistically significant difference was observed iii. Metastatic adenocarcinoma (19/4.0%) between lower class to middle class and higher class of all society. iv. Neuroendocrine (4/0.8%) No difference was observed between the various ethnic groups. Conclusion: Socio-economic factors such as diet and nutrition Conclusion: Our study showed ulcerative colitis as the com- directly affects bone growth and hence bone age. The results of monest premalignant lesion and grade II adenocarcinoma the fusion of the medial end of the clavicle are not affected by ethni- most common malignancy of intestine. city. More studies should be conducted in various parts of country COMPARISON OF ANTIBACTERIAL ACTIVITY OF PUNICA to make a natural standard in setting up uniform criteria for as- GRANATUM AND SYZYIUM CUMINI LEAVE EXTRACTS sessing age at or above 21 years. Huma Urooj1and Aliya Hayat1 CENTRATHERUM ANTHELMINTICUM (KALIZERI) MINIMIZE 1Department of Microbiology, Jinnah University for Women, Kar- THE RISK OF CHEMICALLY-INDUCED HEPATOTOXICITY IN RATS achi, Pakistan. Rabbiya Usmani1, Sumera Rais2, Syed Shabib Zaidi1and Shamim A Introduction and Objectives: Plants are the richest source of Qureshi1 phytochemicals, not only protecting the plants itself but also 1Department of Biochemistry, University of Karachi, benefits the mankind. Punica granatum commonly known as po- Pakistan,2Department of Biochemistry, Federal Urdu University of megranate and Syzyium cumini known as Jamum are well known Arts, Science and Technology, Karachi, Pakistan. for its phenolic compounds having antioxidant potential. They are Introduction and Objectives: Carbon tetrachloride (CCl4) is a used in traditional medicine for the treatment of diabetes, diar- cleaning agent used in industry. Its vapours are injurious for many rheal diseases, antiparasitic and antibacterial infections. body tissues especially liver. The study was designed to evaluate Methods: The aim of the study was to identify the anti- the hepatoprotective effect of ethanolic seeds extract (ESEt) of microbial potential of two leaf extracts of Punica granatum and Centratherum anthelminticum (kali zeri) in CCl4-induced hepa- Syzyium cumini. fresh leaves were washed and ground with distil totoxic rats. water. The extract was obtained which was filtered by Whatman Methods: Rats were made hepatotoxic by intraperitoneal in- #1 filter for the removal of particals. The antimicrobial activity was jection of CCl4 (3 ml/kg) on 3rd and 5th day of trial and divided performed by agar well diffusion method against E.coli, Bacillus into hepatotoxic control (distilled water 1 ml/kg), positive control subtilis, Klebsiella oxytoca, Salmonella typhi, Micrococcus leutus, (silymarin 100mg/kg), and test (ESEt 800 mg/kg) groups. Normal Proteus mirabilis, Staph aureus and Candida albicans. PBS was control rats were also run. After 24 hours of last dose of CCl4, body taken as negative while streptomycin as positive control. weights (BW) of all rats were recorded, sacrificed them to collect Results: It was found that the extract of Punica granatum was blood and serum to analyze liver-specific parameters. In addition, inhibitory to all microorganisms. The highest zone of inhibition livers were dissected out carefully to estimate lipid peroxidation was observed against Proteus mirabilis and E.coli (25mm). Leaf (LPO). extract of Syzyium cumini do not show any antimicrobial activity. Results: ESEt (600 mg) not only significantly decreased the Conclusions: Pomegranate leaf extract can be effective against percent reduction in BW of test rats but also improved the levels of GIT infections. Jamun, as reported earlier, is well known to control alanine transferase (ALT), asparate transferase (AST), alkaline diabetes cannot be used to control infections. 210 N.A. Siddiqui, A. 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IDENTIFICATION, DIFFERENTIATION AND SENSITIVITY PAT- Naheed A Irfan1*, Sumaira Javed1, Justina Abel1and Syeda Gufrana TERN OF HAEMOPHILUS INFLUENZAE AND HAEMOPHILUS Nadeem1 PARAINFLUENZAE 1Jinnah University for Women, Karachi, Pakistan. Naheed A.Irfan1*, Maira Khan1, Tehreem Gul1, AmnaShafique1, Introduction and Objectives: Candida a diverse group of fun- Syyada Ghufrana Nadeem1 gus has bacterial and fungal properties and 80% commensals of 1Jinnah University for Women, Karachi, Pakistan. normal human body i.e C. albicans and C. tropicalis are involved in Introduction and Objectives: H.influenzae is a pleomorphic various diseases like oral thrush, vulvovaginal candidiasis etc. Gram-negative coccobacillus that is isolated predominantly from when they get opportunity and they now are considered as the the respiratory tract and this opportunistic species may cause third most pathogenic strain causing various diseases in human. systemic infections such as epiglottitis, meningitis, bacteremia/ The objective is to compare the virulence properties of different sepsis, bronchitis and chronic obstructive pulmonary disease and Candida spp. otitis in young children. H. parainfluenzae, in contrast, is a sa- Methods: 126 sputum samples were collected from May to prophyte that colonizes the upper respiratory tract but it hardly August 2015 from hospitalized patients. Samples were inoculated causes respiratory tract infections and only occasionally infectious on SDA. Identification was processed by gram staining, germ tube endocarditis. But many strains are now becoming resistant to beta test, Chlamydospore detection and CHROM agar. Phospholipase, lactam antibiotics. We aim to test the sensitivity pattern of Hae- proteinase and hemolytic activity were delibrated in them. Sta- mophilus influenzae and Haemophilus parainfluenzae. tistical analysis was also performed using SPSS version 17. Methods: Total 45 sputum samples were analyzed collected Results: 51% males and 48% female were found to be infected from different hospitalized patients. Specimens were inoculated with Candida spp. The most prevelant organism was C. albicans on blood agar plates and incubated in 5-% CO2 at 370C for 24-72 (87.5%) followed by C. glabrata (8.56%) and C. tropicalis (3.08%). hours.Gram stain were perform for direct examination and dif- Phospholipase activity (45.9%), proteinase (30.1%) and haemolysin ferentiation was done on the basis of availability of hemin(X) and activity (19.9%) was shown by C.albicans. Phospholipase and pro- NAD (V) factor, oxidase and slide agglutination test. Antibiogram teinase activity was also observed in C.tropicalis but not in C. was done by CLSI method. Statistical analysis was also performed glabrata. using SPSS version 17. Conclusions: Present study indicate that C.albicans being a Results: H.parainfluenzae showed such sensitivity pattern member of normal flora pose a major threat in immune-compro- AMP75%, AMC80%, CRO80%, SXT42%, C92%, CIP/OFX84% while H. mised patients and majors should be inforced to prevent infections influenzae showed 100% sensitivity towards AMP, AMC, CRO, SXT, with Candida. C and 92% sensitivity against CIP. STRUCTURAL INVESTIGATION OF NEISSERIA MENINGITIDIS Conclusions: Proper diagnosis and selective use of antibiotics PATHOGENIC FACTORS; A KEY STEP TOWARDS DRUG DESIGNING can minimize the resistivity of these organisms. Sana Aurangzeb1, Mehwish Hamid1and Yasmeen Rashid1 ASSOCIATION BETWEEN INTERLEUKIN 6 GENE POLY- 1Department of Biochemistry, University of Karachi, Karachi, MORPHISM AND HUMAN PAPILLOMA VIRUS INFECTION IN Pakistan. ORAL SQUAMOUS CELL CARCINOMA PATIENTS Introduction and Objectives: The Gram negative human pa- Mehwish Zafar1, Naila Hadi1, Saeeda Baig1and Nosheen Zehra1 thogenic bacterium Neisseria meningitidis is responsible for 1Ziauddin University, Clifton Campus, Karachi, Pakistan. causing meningitidis and septicemia worldwide. A substantive Introduction and Objectives: Human papilloma virus (HPV) question in bioinformatics analysis of bacterial genome is to as- and IL 6 gene polymorphism are proven independent risk factors cribe a three dimensional structure as well as a biologic role to all for development of Oral Squamous Cell Carcinoma. The objectives the coding regions. Here we performed structural bioinformatics of this study were to find out frequency of IL6 gene polymorphism analyses of important Neisseria meningitidis pathogenic factors and HPV and to evaluate any association between HPV and IL6 involved in protein biosynthesis including methionyl-tR-NA syn- gene polymorphism in OSCC patients and its effect on disease thetase, 16SRNA methyltransferase, translation elongation factor- prognosis. Tu and putative RNA methylase. Methods: In this cross-sectional study 140 OSCC patients (104 Methods: Homology modeling of these important drug targets males and 36 females), aged 18yrs and above were selected. De- was carried out with better templates by MODELLER software and tailed questionnaire was filled followed by sample collection. After evaluated by Prosa and Procheck standalone softwares. DNA extraction PCR analysis for HPV and RFLP for IL6 gene poly- Results: The study provided detailed structures of important morphism was carried out. proteins required for the pathogenesis of this organism. Results: Mean age of the patients was 43.5±11.84 years (range Conclusions: In future, these investigations will help in re- 31-40 years). Majority (n=45; 32.1%) belonged to Urdu speaking ceptor-based drug designing against meninigococcal infections. ethnic group and were habitual eaters of Pan (n=87; 62.1%) and EVALUATION OF ANTIMICROBIAL ACTIVITY OF ALOE VERA Gutka (n=82; 58.6%). The most common site of OSCC was buccal Iqra Waqar1and Aliya Hayat1 mucosa (61.4%). Most of the patients presented with grade II (55%) 1Department of Microbiology, Jinnah University for Women, Kar- and late stages (stage III & IV) (55.7%) of OSCC. Out of 140 samples, achi, Pakistan. 12 (8.6%) tested positive for HPV gene with the following pattern Introduction and Objectives: Aloe Vera is a cactus, belongs to of IL 6 gene polymorphism; GG (46.4%), GC (39.3%), CC (14.3%). A the family Liliaceae, and is well known for its use as traditional significant association was observed between stages III & IV of medicine. It is use for gastrointestinal disorders, stimulates body’s OSCC and IL6 genotypes GC (P=0.001) and CC (P=0.002). Also immune response, for treatment of burns, eczema, psoriasis, to strong positive association (P=0.003) was found between HPV and heal skin infections. Aloe Vera is known to hold therapeutic IL 6 (P=0.007) CC homozygote genotype. properties due to bioactive components present in it for both in- Conclusions: This study finds an association between HPV and fectious and non-infectious diseases. IL6 gene polymorphism in late stage OSCC patients suggesting Methods: Antimicrobial activity of Aloe vera was investigated rapid and aggressive progress of oral carcinogenesis. against Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, COMPARISON OF VIRULENCE FACTORS OF ISOLATED CANDI- Micrococcus, Klebsiella pneumonia, Proteus, Staphylococcus aur- DA SPP FROM LOWER RESPIRATORY TRACT INFECTIONS IN eus, Salmonella typhi and Candida albicans. The organisms were HOSPITALIZED PATIENTS isolated from clinical samples and identified by conventional N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 211 methods. Susceptibility test was carried out by agar well diffusion UNDERSTANDING DRUG RESISTANCE IN BREAST CANCER method. Aloe vera used as methanolic extract and pure gel. All Umber Saeed1and Usman Choudhary2 experiments were performed in triplicate. 1Dow College of Pharmacy, DUHS, Karachi, Pakistan,2London Results: Zone of inhibition was observed in all forms of Aloe PGDE, Scotland, UK. vera. The maximum zone of inhibition was found in E.coli and Introduction and Objectives: Breast cancer is one of the followed by Pseudomonas aeruginosa, Salmonella typhi, S aureus leading cause of death worldwide. In Pakistan, prevalence of this and Bacillus subtilis showed least zone of inhibition. It was also ailment is highest amongst all types of cancer i.e. 38.5%. Major found that inhibitory affect against Candida albicans was also clinical setback is drug resistance in breast cancer. Metabolomics is maximum. an emerging field that utilizes information of cellular biochemistry Conclusions: The result of this study would tend to give cre- for the early detection, diagnosis and establishment of predictive dence to the use of Aloe Vera for the formulation of certain biomarkers of breast cancer. This review highlights potential me- compounds that would coin new, effective and more potent an- tabolomic applications to pharmacology and clinical timicrobial drugs to combat pathogenic microorganisms. pharmacology. BIOINFORMATICS ANALYSIS OF HISTONE-LYSINE N-METHYL- Methods: The methodology is based on inclusion exclusion TRANSFERASE - SETD1A GENE IN ASSOCIATION WITH criteria. Literature survey, and questionnaire were included while SCHIZOPHRENIA clinical trial was excluded. This report provides a review of 12 Zoobia Anis1, Hajira Karim1and Afshan Zeeshan Wasti1 articles out of few were excluded. 1Department of Biochemistry, Jinnah University for Women, Kar- Results: According to the survey the average response rate of a achi, Pakistan. cancer drug is the lowest at 21%, suggesting that 74% of patients Bioinformatics methodologies have made it possible to profile with cancer are over-dosed. While according to an international the global composition of tissue or organelle at specific time point study, 40%–50% of breast tumors will display acquired resistance. or under particular developmental or disease state. A genetic Conclusions: When specific therapies are chosen on the basis variant of SETD1A has yielded significant genome-wide associa- of a patient's metabolomics profile, it will give rise to customized tion with schizophrenia, suggesting that this SETD1A plays a key medicine and personalized tailored treatment. Using high- role in its etiology. The SETD1A gene plays a part in chromatin throughput information using metabolomics to clinical diagnosis modification - an important cellular process that reduces the size and treatment can help accelerate the patient safety, quality of life of DNA so it can fit inside a cell and regulate gene expression. It and survival rate by identifying pathways involved in drug encodes a catalytic subunit of an enzyme that adds methyl groups resistance. to histone proteins - genes for transcription. ABERRANT PROTEIN S-NITROSYLATION IN HYPERTENSIVE The present study is based on computational analysis of SET- AND DIABETIC-HYPERTENSIVE PATIENTS: NEW PERSPECTIVE D1A gene exploring molecular networks of interacting targets by Erum Zafar1, Ayesha Khan1, Beena Hasan1and Nikhat Ahmed1,2 using advanced bioinformatics tools such as, STRING’s, KEGG, Re- 1Neurochemistry Research Laboratory, Department of Biochem- actome and BioGrid databases that may provide insights into the istry, University of Karachi, Pakistan,2Research Department, Ziauddin biological processes underlying schizophrenia. University, Karachi, Pakistan. FREQUENCY OF HEPATITIS B AND C IN DIFFERENT AREA OF Hypertension and diabetes are the risk factor of morbidity and CATCHMENT CENTER IN GADAP TOWN, KARACHI, PAKISTAN mortality. The prevalence of hypertension is expected to increase Aliya Zaman1, Nazish Jaffer2and Rubina Ghani3 in the upcoming years. The incidence of diabetes is two fold higher 1,2Department of Pathology, Sir Syed Medical College, Karachi, in patient with hypertension. The co-existence of diabetes and Pakistan2Jinnah Sindh Medical University, Karachi, hypertension causes impaired renal function, the development of Pakistan,3Department of Biochemistry, Jinnah Medical and Dental diabetic retinopathy, cardiac problems, and the development of College and Pathological and Molecular Laboratories, Karachi, cerebral diseases. Besides these disorders hypertension and dia- Pakistan. betes are involved in many other devastating disorders. Protein S- Introduction and Objectives: Frequency of Hepatitis B Surface nitrosylation conveys a large part of the ubiquitous effect of nitric Antigen (HBs Ag) and Hepatitis C Virus Antibodies (anti-HCV) oxide on cellular signal transduction, accumulating evidence in- among population of different catchment center in Gadap area dicates important roles for S-nitrosylation both in normal phy- with their low social economic status. We aim to study the fre- siology and in a broad spectrum of human diseases. Dysregulated quency of Hepatitis B Surface Antigen (HBs Ag) and Hepatitis C S-nitrosylation has been implicated as a cause or consequence of a Virus Antibodies (Anti-HCV) in different catchment center of Ga- broad range of diseases, including asthma, cystic fibrosis, Parkin- dap town in 12 kilometer from Baqai Medical University. The de- son disease, heart failure, and stroke. The role of nitrosylases and sign of study will be cross sectional descriptive study. It was held denitrosylases in governing levels of S-nitrosylation under both in the Pathological and Molecular Laboratories, Karachi, during the physiological and pathophysiological conditions is increasingly period October, 2011 to May 2012. appreciated. The purpose of study is to identify s-nitrosylation in Methods: A total of 496 samples were collected. Serum was hypertensive and diabetic hypertensive serum samples. We take tested for Hepatitis B surface antigen and for Hepatitis C virus serum sample of hypertensive subjects (n=10), diabetic hy- antibodies by ELISA. The results were subjected to chisquare pertensive subjects (n=10) and normotensive subjects (n=10). analysis for determination of statistical difference between the Total protein was quantified using Bradford assay. SDS PAGE was values among different categories. done after dilution of sample followed by western blotting. The Results: Among 496 patients 91 (18.3%) were positive for HB s results we obtained from the above mentioned methods showed Ag and 171 (34.4%) were positive for Anti-HCV. The frequency of that s-nitrosylation is decreased in hypertension and slightly in- HB s Ag was seen too low as compared to anti-HCV. creased in diabetic hypertension. Conclusions: High frequency of HCV infection needs im- STRUCTURAL BIOINFORMATICS ANALYSIS OF TRANSCRIP- plementation of strict screening policy for donors and public TION FACTORS INVOLVED IN NEISSERIA MENINGITIDIS PATHO- awareness campaigns about preventive measures to reduce the GENESIS; A LEAP TOWARDS ANTIMENINGOCOCCAL DRUG spread of this infection as well as other transfusion transmissible DISCOVERY infections. Mehwish Hamid1, Sana Aurangzeb1and Yasmeen Rashid1 INNOVATIVE APPROACHES IN METABOLOMICS FOR 1Department of Biochemistry, University of Karachi, Karachi, 212 N.A. Siddiqui, A. 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Pakistan. Areeba Asghar1, Saeeda Baig2, Asghar Ali1and Muhammad Asif Introduction and Objectives: Neisseria meningtidis is a viru- Memon1 lent pathogen causing meningtitis as well as life-threatening 1Department of Physiology, Ziauddin University, Karachi, septicaemia throughout the world. Unfortunately, any vaccine or Pakistan,2Department of Biochemistry, Ziauddin University, Karachi, drug has not yet been developed against N.meningtidis serogroup Pakistan. B. Functional genomics strategies have been adapted to study the Introduction: Salivary flow and composition alters under de- growth and pathogenesis of Neisseria meningtidis in which a li- leterious chemical irritants present in chewable tobacco. Apart brary of 2850 insertional mutants was analysed and 73 genes were from inflammation and deterioration in health of oral mucosa it identified in N.meningtidis genome responsible for causing can also affect the major and minor salivary glands and cause a disease. decline in salivary flow rate. The objective of the study was to find Methods: We have selected the proteins (gene products) i.e., out alteration in salivary parameters that lead to oral lesions Dead box RNA-Helicase, Polyribonucleotide nucleotidyl-transfer- among chewable tobacco users ase PNPase and Ribonuclease-III involved in transcription for their Methods: A total of 354 healthy male subjects, consuming any detailed structural analyses. These proteins are also involved in form of chewable tobacco product, belonging to low socio- RNA-degradosome assembly. economic areas of Karachi were selected for this cross sectional Results: Different bioinformatics strategies were applied and study. A questionnaire was used to collect demographic data and the homology models of these proteins were built using protein details of chewing habits (using since, pack/day, duration of ex- structure-modelling program MODELLER and the models were posure etc.). Resting saliva of every subject was collected for 5min evaluated using PROSA and PROCHECK software, as well as active and RSFR was expressed in ml/min. Salivary pH was determined sites are also predicted. by using pH strips (pH 0-14). Oral examination was done for the Conclusions: Since these proteins are potent dug targets, presence or absence of oral lesions. Data was analyzed on SPSS therefore their structural identification will prove to be a break- version 20. through in drug discovery. We hope that our study will grease the Results: Out of the 354 subjects included, 27.4% consumed wheels for drug designing against lethal meningococcal disease. gutka, 24.3% niswar, 24.3% paan and 24% were multiple users. ROLE OF RAUWOLFIA SERPENTINA EXTRACT ON THE MOD- Mean Resting Salivary Flow Rate (RSFR) was 0.52±0.34 ml/min, pH ULATION OF IMMOBILIZATION STRESS-INDUCED BEHAVIORAL 6.58±0.78 and 27.1% (n=96) had oral lesions. Highest frequency of DEFICITS AND ADAPTATION TO STRESS IN RATS oral lesions was found among subjects who had hyposalivation Erum Shireen1, Wafa Bint E Ali1and Darakshan J Haleem1,2 (40%) and those having acidic pH (40%). A significant decrease in 1Department of Biochemistry, Neurochemistry and Biochemical RSFR and pH and increase in frequency of oral lesions is observed Neuropharmacology Research Laboratory, University of Karachi, with increased duration of exposure, duration of usage and in- Pakistan,2Neuroscience Research Laboratory, Dr Panjwani Center for creased number of tobacco packs consumed per day. Molecular Medicine and Drug Research, University of Karachi, Conclusion: Increased frequency and recency of chewable to- Pakistan. bacco use leads to decrease in RSFR and pH and hence increase in Introduction: Stress is the major precipitating factor in the frequency of oral lesions. onset of depression, anorexia nervosa, diabetes and obesity. The SPEECH ASSESSMENT THROUGH THETRI-POSITION ARTICU- goal of present study is to monitor the behavioral and biochemical LATION ANALYSIS FOR BILINGUALS (TAAB) effects of post stress administration of Rauwolfia serpentina ex- Amina Asif Siddiqui1 tract on adaptation to immobilization stress in rats. 1College of Speech Language and Hearing Sciences, Ziauddin Methods: The plant extract (30 mg/kg) was orally administered University, Karachi, Pakistan. after termination of immobilization stress daily (2 hours) for 5 Introduction: Bilingualism is regarded as a cognitive feat by days to monitor any change in behavioral activities. Effects of today's linguists. The acquisition of more than a single language Rauwolfia serpentina extract on immobilization stress induced manifests the miracle of the human brain in terms of meta lin- deficits of food intake and body weight were determined for an guistic (meta phonological) skills, as it stores the form and content understanding of the relationship between stress tolerance and of multiple languages and exhibits their use in socially and lin- behavioral changes. We also tested effects of Rauwolfia serpentina guistically appropriate contexts. extract on endogenous leptin and glucose levels in unstressed and Methods: Pakistan, like most Asian countries has a pre- stressed animals to explore the possible role of HPA axis in the dominantly bilingual fabric. Little children, regardless of their so- modulation of stress-induced behavioral deficits and adaptation to cioeconomic status are most often exposed to their native lan- stress. guage (L1) and to a second language (L2) within the critical period Results: The present study showed that Rauwolfia serpentina (0-3years), leading to simultaneous bilingualism. When they are extract can blunt stress-induced anorexigenic as well as anxio- exposed to L2 after the acquisition of L1 we call it sequential bi- genic-like effects. Moreover, Rauwolfia serpentina extract reversed lingualism. Some children may have L3 in their linguistic re- adverse effects of stress and facilitate adaptation to an un- pertoire which may be the additional language such as the lan- controllable stressor by reducing stress perception. guage of instruction at school. The development of articulation Conclusion: The present study shows that immobilization and phonological skills in children usually reaches maturation stress induced decreases in food intake, body weight as well as around the chronological age of 6 years. Children pick up several behavioral deficits were reversed by Rauwolfia serpentina extract sounds of their language and perfect them gradually using ap- suggesting anxiolytic like profile of drug. This effect of Rauwolfia proximations that sound closer and closer to the target sound, serpentina extract can be explained in terms of additive effects of acquiring all sounds with the culmination of early childhood. stress on serotonin (5-hydroxytryptamine; 5-HT) neurotransmis- Results: Children may present with unclear speech, in the ab- sion particularly via postsynaptic 5HT2C as well as 5HT1A re- sence of an organic etiology (dyslalia or phonological disorders) or ceptors. It is therefore suggested that post stress administration of because of a known organic cause such as cleft lip/palate, mal- Rauwolfia serpentina extract provides an innovative approach for occlusions, hearing loss, dyspraxia or dysarthria, must be assessed the treatment of stress related disorders. for their ability to acquire phonemes of all the languages in their ALTERATION IN SALIVARY PARAMETERS LEAD TO ORAL LE- repertoire, because the brain stores auditory images of all sounds SIONS AMONG CHEWABLE TOBACCO USERS and uses them appropriately when needed. N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 213

Conclusions: The Tri-position Articulation Analysis for Bilin- 1Ziauddin University and Hospital, Karachi, Pakistan. guals is a tool for testing the articulation and phonological skills of Introduction and Objectives: Vitamin D deficiency may affect bilingual children aged 0-6 years, that uses culturally and lin- the mother and its new born health. The aim is to detect the fre- guistically appropriate picture stimuli for children exposed to quency of vitamin D deficiency in pregnant women and their Urdu, Sindhi, Punjabi, Pushtu, Hindi and English. The test assesses newborn of Karachi at tertiary care hospitals, and correlating spontaneous elicitation of the sounds in the child's lexicon, along maternal vitamin D with cord blood. with stimulability and also assesses the function of the speech Methods: This cross-sectional study was conducted on 50 mechanism for non-speech functions. women in labor presenting with a singleton term pregnancy at STABILITY STUDIES OF METFORMIN HYDROCHLORIDE, AM- tertiary centers in Karachi. Data was recorded on a special pro- LODIPINE BESYLATE AND CIPROFLOXACIN HYDROCHLORIDE forma, maternal blood was taken before delivery and cord blood TABLETS UNDER SPECIFIED CONDITION OF TEMPERATURE AND was taken at delivery. All blood samples were analyzed for 25- RELATIVE HUMIDITY OF PAKISTAN hydroxy vitamin D levels. Qurratul ain Leghari1, Sohail Hassan2and Iqbal Ahmed3 Results: The mean vitamin D levels were 24 ng/mL for the 1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, mothers and 20 ng/mL for the newborns. Vitamin D sufficiency Ziauddin University, Karachi, Pakistan,2Department of Pharmaceu- was noted in 11 (22%), insufficiency in 16 (32%), and deficiency in tical Chemistry, Faculty of Pharmacy, University of Karachi, Karachi, 23 (46%) of the 50 mothers whereas sufficiency and deficiency, Pakistan,3Baqai Medical University, Karachi, Pakistan. were noted in 6 (12%) and 44 (88%) of the newborns respectively. A pharmaceutical formulation ensures safety, protection, pro- There was a positive correlation between the vitamin D levels in curement and preservation of human health. Therefore, pre- maternal and cord blood (r=0.03; P=0.003). Maternal vitamin D servation of its physical and chemical stability throughout the levels were significantly affected by sunlight exposure (P=0.007) period of its shelf life has always been the matter of prime concern and quality of diet (P=0.01). for a pharmaceutical manufacturer. A good formulation is one that Conclusion: Vitamin D deficiency is high among pregnant ur- when packaged in a suitable container, (which provides maximum ban Pakistani women and their newborns. This public health protection to its contents), confirms the absolute delivery of its problem needs urgent attention. active ingredient in its original form to its consumer. To determine ARE YOU PREPARED TO CARE YOUR LOVED ONES: A CROSS the suitability of blister strips for packaging of Metformin HCl, SECTIONAL STUDY TO ASSESS THE LEVEL OF PREPAREDNESS Amlodipine besylate and Ciprofloxacin hydrochloride tablets, we AMONG CAREGIVERS OF CHRONICALLY ILL PATIENTS IN A TER- maintained a stability profile for the changes in physical appear- TIARY CARE HOSPITAL AT KARACHI, PAKISTAN ance and loss of potency of these medicines for a period of 2 shelf Shireen Arif1 life years. Tablets were subjected to the specified conditions of 1College of Nursing, Ziauddin University, Karachi, Pakistan. 30oC and 65% RH for 24 months and samples were withdrawn at Introduction: Family caregivers are also known as informal different time intervals. Different tests and assays were conducted caregiver. They provide all level of care at home to their loved ones on these samples showed that under these specified conditions most often equivalent to the professional care providers, such as the efficacy of these tablets in the blister strips retain for less than providing physical care, giving medications recognizing early signs 2 shelf life years which is less than the expected shelf life of 3 of severe conditions, changing dressings etc. While providing care years. to their loved ones most of them feel unprepared in terms of lack RETROSPECTIVE ANALYSIS OF MATERNAL MORTALITY AT of knowledge and skills. This unpreparedness may result in the ZIAUDDIN UNIVERSITY HOSPITALS, FROM 2012- 2014 role strain and have may psychological consequences for the fa- Chaudhry Shama1 mily. Nurses as a primary care provider could assess the caregiver 1Ziauddin University, Karachi, Pakistan. preparedness and may educate them according to the patient’s Introduction and Objective: To determine the frequency of need to reduce the extra burden and the role strain. maternal mortality and identify the causes with different age Method: The self-administered instrument regarding the pre- groups, parity and gestational ages in Ziauddin university hospitals paredness for care giving scale by Archbold (1990) has been uti- in 4 campuses, from 2012- 2014. lized. The scale has eight items to identify the preparedness of Methods: A descriptive study conducted at Gynecology & Ob- caregiver in multiple domains. The responses are rated on five stetrics units of Ziauddin university hospitals with retrospective point scale with scores ranges from 0 (not at all prepared) to 4 review of validated records of hospital registers in 3 years from (very well prepared). The higher score means caregiver feels more January 2012 to December 2014. Convenience sampling used to preparedness and vice versa. 50 caregivers of chronically ill pa- access the records. A self-structured proforma designed to collect tients were selected at a tertiary care hospital, which were caring data on variables (age, parity, gestational age, cause of death, for their loved ones for about six months or above. Written con- condition at the time of admission, duration of stay in hospital sent was obtained from the participants. before death, cause of delay). Result: 50 caregivers were recruited to fill the form from which Results: During 3 year period from January 2012 to December 26 were female and 24 were male. All of the participants were 2014 there were 32 maternal deaths. Total number of births were between 18 years and 60 years of age. About 68% caregivers score 14219 and 14184 were live births. Still births were 33. Eclampsia between not too well prepared and somewhat well prepared. (21.8%) and haemorrhage (21.8%) were the leading cause of ma- Whereas, only 32% score between somewhat prepare and pretty ternal mortality followed by those with puerperal sepsis (12.5%) well prepared. and others. MMR (maternal mortality ratio) was 226 per 100000 Conclusion: In conclusion, most of the caregivers score are live births. somewhat prepared which is an alarm for nurses and all health Conclusion: Maternal mortality was high with leading causes care providers to work to reduce their role strain and provide as hemorrhage, eclampsia, and sepsis in ages of 20-30 years and sessions according to the need of the caregivers. Nurses could ar- parity 1-4. Mostly deaths were due to delay in getting the women range teaching sessions which could be helpful for the caregivers to the health facility. to do their role effectively. VITAMIN D DEFICIENCY IN PREGNANT WOMEN AND THEIR ABO AND RHESUS BLOOD GROUP DISTRIBUTION IN RE- NEW BORN SIDENTS OF KARACHI Shazia Sultana1 Sumera Shaikh1, Shariq Adnan1, Mustafa Zuberi1and Syed Sohail 214 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219

Abbas1 blood bank and transfusion services as a routine practice. 1Ziauddin University, Karachi, Pakistan. SELF MEDICATION PRACTICE AMONG UNDERGRADUATE Introduction and Objective: Blood is an essential component MEDICAL STUDENTS for human beings and in disease states, it is requested to be Tahira Asad1, Tahira Zamir1, Nasim Karim1and Talea Hoor1 transfused from healthy human donors. But it was continuously a 1Ziauddin University, Department of Pharmacology and Ther- life threatening procedure until the discovery of ABO blood apeutics, Bahria University Medical and Dental College, Karachi, groups. Today committee of League of Nations has recommended Pakistan. A, B, AB & O classification for general use. This study was aimed to Introduction and Objective: Assessment of self-medication determine the frequency of ABO and Rh blood groups. So that practice among medical students is presumed to be of exceptional necessary measures should be taken to maintain the blood pro- importance as in future they have promising role in counselling duct inventory. Frequency of ABO and Rhesus blood group dis- the patients regarding the rational use of drugs. This study was tribution in residents of Karachi. specially designed to evaluate self-medication practice in 3rd year Methods: A study was conducted at Dr. Ziauddin Hospital medical students. Clifton laboratory Karachi, Pakistan over a period of 7 months from Methods: A cross sectional institution based study was con- 1st June until 31st Dec 2014. Healthy blood donors and patients ducted from Jan – March 2013 on 3rd year medical students at requiring blood products of different age groups were employed. Bahria University Medical and Dental College, Karachi, after ap- After an informed consent blood grouping was performed by proval by ERB-BUMDC. Data was collected by questionnaire, filled taking blood samples under aseptic measures in EDTA and Gel on one to one basis after taking verbal consent and analysed on separating serum tubes for ABO & Rh blood grouping by tube SPSS version 16. method. Results: Frequency of self-medication was found to be 90%. Results: Total 1583 individuals included 868 (54.8%) males and Quick relief (38%) and busy schedule (21%) were the main reasons 715 (45.2%) females. O+ve blood group found to be more common provoking self-medication. Common ailments that led to self- group (31.9%) followed by B+ve (31.2%). In Rh system, Rh+ve medication were: headache (95%) and fever (87%). Drug groups (97.2%) were more common than Rh-negative (2.8%) blood groups. frequently used were: analgesics (95%), antipyretics (87%) and Conclusion: This study was taken out to maintain the blood antibiotics (36%). Acetaminophen was the most common analgesic product inventory concerning the management of blood bank and (73%) as well as the antipyretic (88%) while antibiotics were, me- transfusion services for the patient. tro-nidazole (26%) followed by amoxicillin plus clavulanic acid FREQUANCY OF ABO BLOOD GROUP DISCREPANCIES IN TER- (17.3%). TIARY CARE HOSPITAL KARACHI AND THEIR CLASSIFICATION Conclusion: Practice of self-medication is high among medical INTO GROUPS students. Potential problems of self-medication should be em- Sumera Shaikh1, Saba Jamal1and Fatima Mairaj1 phasized to the students. 1Ziauddin Hospital, Karachi, Pakistan. DETERMINATION OF PROSTATE GLAND VOLUME BY ULTRA- Introduction: In developing countries, the blood grouping is SONOGRAPHY AND ITS CORRELATION WITH ANTHROPOMETRIC generally being performed by the forward grouping method only. MEASUREMENTS IN A SUBSET OF KARACHI POPULATION The aim of this study was to find out the frequency of ABO blood Iffat Raza1 group discrepancies in tertiary care hospital in metropolitan city of 1Department of Anatomy, Ziauddin University, Karachi, Pakistan. Pakistan and also to classify these discrepancies on the etiological Introduction and Objectives: To establish the local reference basis. range of prostate volume according to our subset of population. To Methods: Cross-sectional, descriptive type of study was con- correlate prostate volume (PV) with age, body mass index (BMI) ducted at Blood Bank of Dr. Ziauddin Hospital Karachi, Pakistan. and waist circumference (WC). Total 1522 samples were included with exclusion of newborns and Methods: A cross-sectional study with 119 healthy adults aged infants up to the age of 6 months. Blood samples were collected 40-79 years without any prostatic pathology were recruited. The from a peripheral vein by trained phlebotomist and submitted to study population was categorized into 4 age groups(40 - o50 the blood bank department in 2 tubes, purple top vacutainer tube yrs,50 -o60 yrs,60-o70yrs,70-o80 yrs), 3 BMI groups (healthy, containing Di-potassium Ethylene Diamine Tetra Acetic Acid (K2 obese and overweight) and 2 WC groups (o 90cm and 490cm). A EDTA) anticoagulant and red top vacutainer tube containing clot- p-value of o0.05 was considered significant. A total of 130 healthy ted blood. Blood group testing was performed by tube technique. adult volunteers were recruited initially, out of these 20 in- Results: In total 1557 patients (854 Males and 703 Females) dividuals were those whose IPSS was o 8 and prostate volume on from various clinical specialties were included for ABO blood ultrasound was 4 25 ml. These 20 subjects underwent Uro- grouping. The male to female ratio was 1.2:1 (854 males and 703 flowmetry (UFM). After UFM, total 19 subjects were labelled as females) with age ranging from 9 months to 92 Years. ABO dis- healthy adults and 1 subject was excluded from healthy adult crepancies were Positive in 18 out of 1557 Patients (1.1%). Male to group. The sample of 119 healthy subjects were studied sono- female ratio among ABO discrepancies were 1.5:1. There were four graphically for the effects of Age, BMI and WC on prostate volume. groups of ABO discrepancies. The most common group of dis- Results: The mean prostate volume was 21.7±2.2mls, mean crepancies were group I discrepancies consisting of 12 out of 18 body mass index was 28±6kg/m², whereas mean waist cir- Patients (66.7%). Group II discrepancies were consist of 2 out of 18 cumference was 95cm. PV was found to be higher in obese and Patients (11.1%). Group III discrepancies were consisting of 1 out of 490cm waist circumference group. After applying multiple re- 18 Patients (5.5%) and Group IV discrepancies were consisting of 3 gression analysis, waist circumference correlated positively and out of 18 Patients (16.7%). significantly with prostate volume. UFM showed negative corre- Conclusion: The forward and reverse grouping are the essen- lation with IPSS and PV. tial parts of blood group testing for assigning the correct ABO Conclusions: Mean prostate volume in our studied population blood group to the individual. ABO discrepancies i.e. forward and was smaller than that of many western populations. Our study has reverse grouping are not matches to each other can cause severe proved that central obesity is the most important factor influen- transfusion reactions which can be fatal for life. Furthermore, to cing prostate volume. ensure that correct ABO blood product to be transfused for saving ASSOCIATION OF KISSPEPTIN, ENDOMETRIAL RECEPTIVITY life. Therefore reverse grouping should be strictly adopted in every AND UNEXPLAINED INFERTILITY N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 215

Rehana Rehman1, Zehra Jamil1, Syeda S Fatima1, Sara Arif2and the decidualized endometrium act as a biosensor of good quality Faiza Alam1 embryos, if it is faulty, may lead to implantation of embryos re- 1Department of Biological and Biomedical Sciences, Aga Khan sulting in miscarriage. Other factors implicated in the pathophy- University, Karachi, Pakistan,2Civil Hospital, Karachi, Pakistan. siology of miscarriage are Systemic and placental oxidative stress. Introduction and Objectives: To relate serum Kisspeptin levels Vascular endothelial damage, abnormal vascularity of placenta and with pregnancy outcome after intra cytoplasmic sperm injection immunologic reactions have been proposed to play some role in (ICSI). causing recurrent miscarriage. Methods: In the cross sectional survey carried at Australian The objective of this review is to discuss the causes of recurrent Concept Infertility Medical Centre from June 2014 to June 2015, miscarriage, based on the published research articles. 176 females with 20 to 42 years of age and with regular menstrual PREVALENCE AND PREVENTIVE MEASURES OF PROSTATE cycles were included for ICSI. Patients with uterine fibroids and CANCER: A RANDOMIZED STUDY IN PUBLIC AND PRIVATE metabolic disorders were excluded. Down regulation of ovaries HEALTH CARE SECTOR was followed by controlled ovarian stimulation, ovulation induc- Somia Gul1, Umm-e-Aimon1and Maria Ayub1 tion, oocyte retrieval, microinjection and embryo transfer. Serum 1Faculty of Pharmacy, Jinnah University for Women, Karachi, samples for estimation of Kisspeptin, was done on OI day and Pakistan. endometrial thickness was measured. Based on beta hCG results Introduction and Objectives: Prostate cancer accounts for were categorized into group A, non-pregnant with beta hCG o25 abnormal or uncontrolled division of cells of prostate in male. It is m I U/ml II, and group B, clinical pregnancy with beta hCG45m a slow growing cancer which is usually localized rarely metasta- IU/ml. sized. The growth rate varies from slow moderate to high. It Results: Kisspeptin levels were significantly higher in the mostly occurs in old age and depends on many other factors like Group B versus Group A (po0.001) independently associated with weight, height, diet, habit, environment, genetics, and sexual dis- positive pregnancy (r = 0.388; p o0.001), and endometrial ease. This study aims to evaluate occurrence, causes, problems thickness (r = 0.294; p = 0.05) irrespective of the age and BMI of associated with disease and treatment of this cancer in view of the the subjects. fact to evaluate consequences responsible to contribute carcinoma Conclusion: Kisspeptin is a positive pregnancy marker in fe- of prostate. males after ICSI as a result of its effects on oocyte maturation and Methods: For this purpose randomized study conducted at endometrial thickness. public and private health care sector (n= 100 patients of prostate EFFECT OF LONG HOUR SHIFTS ON MEDICAL INTERNS AND cancer). RESIDENTS, AND THE STRATEGIES ADOPTED TO COPE WITH Results: The Gleason score are used to diagnose prostate can- WORK-RELATED STRESS cer, 10% of patients have score 5 (4+1), 40% of patients have score 7 Maryam Kazim Dattoo1, Nimra Shah1and Aliza Fatima1 (4+3) or (3+4), 44% of patients have score 9 (5+4), 6% of patients 1Ziauddin University, Karachi, Pakistan. have score 11 (5+6) which shows that mostly prognostic grade II, Introduction and Objectives: To assess the impact of long hour III IV, V occur. Mostly traditional prostectomy is used as a treat- shifts on performance and health of medical interns and residents ment and 78% of the patients got benefit from it. and strategies adopted to cope with stress. Conclusions: This study concludes that the prostate cancer Method: This cross-sectional study was conducted from Jan- may occur due to age factor (old age), fatty diet, tobacco or alcohol uary to July 2015. A sample of 200 interns and residents was taken intake. Finding of cancer at score 5,7,9,11 and associated compli- from Ziauddin Hospital Clifton, Kemari and North Nazimabad. Data cations are hypertension, urinary tract infection, kidney stones. was collected through a self-designed questionnaire and stress KNOWLEDGE AND ATTITUDE TOWARDS MENOPAUSE IN levels assessed through General Health questionnaire (GHQ-12). WOMEN AGED 45-60 YEARS Data was analyzed through SPSS v 20. Chi-square and Paired T test Rehana Yasmeen1, Rubina Hussain1, Shama Chaudhry1and Aliya was applied. Waheed1 Results: The mean age of participants was 27+ 2.3 years. 1Ziauddin Hospital North Nazimabad, Karachi, Pakistan. Average sleep hours when doing call were 3.06 +1.47. Work per- Introduction and Objectives: Menopause has significant phy- formance was below average in 62 % of the population. Stress was sical, mental, social and psychological effects on women’s life and found in 34% of study population. Increased tea intake was the prior knowledge about these changes help them to cope with stress coping strategy adopted by 72 %. There was a significant these changes with greater readiness. Educating women regarding association between stress level, gender and personal habits. menopausal changes would definitely equip them better to toler- (p=o0.05). A significant difference was found between cigarette ate these menopausal symptoms. and energy drink intake before and after starting job (p=o0.05). Methods: A cross sectional study based on sample of con- Conclusion: High stress may have negative effects on health venience was conducted at the outpatient department of Ziauddin and performance. Hence, medical interns and residents need Hospital, Karachi from 1st July 2014 to 31st January 2015. The support to cope with stress. study participants were 250 menopausal women between the RECURRENT MISCARRIAGE AND ASSOCIATED FACTORS ages of 45-60 years. Data was collected by means of a ques- Rehana Yasmeen1and Rubina Hussain1 tionnaire, which consists of three parts containing questions about 1Ziauddin University, Karachi, Pakistan. demographic information, knowledge and attitude. The incidence of recurrent pregnancy loss in women of re- Results: The mean age of respondents in the current study was productive age group is 0.5%–3%, and idiopathic causes accounts 51.9 +/- 4.2years. The mean age at menopause was 45.9 for 50%–60% of recurrent pregnancy losses. Approximately 30%– +/-3.8years. As far as the knowledge about menopause is con- 50% of conceptions end in spontaneous miscarriage before the cerned 4.8% had weak, 71.2% had moderate and 24% had good completion of first trimester. Miscarriages mostly occur at the time knowledge about menopause phenomenon. Majority of re- of implantation. The causes of recurrent pregnancy loss are many spondents had positive (72.4%) attitude towards menopause and including parental chromosomal abnormalities, maternal throm- 27.6% had negative attitude towards menopause. bophilias, immunologic causes and endocrine disorders. Recurrent Conclusions: Appropriate knowledge and positive perception pregnancy loss is an extremely distressing clinical problem for towards menopause is very important to help women to cope with women as well as health professinals. According to current studies menopause transition and can help in the development of 216 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 appropriate programmes to promote women health. included. The prescription order review prior to dispensing was SOCIAL SUPPORT AS A DETERMINANT OF INTERNALIZED made by experienced and specialized pharmacists in pediatric STIGMA AND SELF-ESTEEM IN PERSONS WITH SCHIZOPHRENIA pharmacotherapy in a research setting. Prescription order in error Umara Rauf1and Uzma Ali1 was marked if not in compliance with standard pediatric refer- 1Institute of Clinical Psychology, University of Karachi, Pakistan. ences, dosing guidelines and antibiotic prescription guide adopted Introduction and Objectives: Social support works as a buf- in the hospital. As per predefined criterion in our study, errors fering factor in psychotherapeutic treatment for the diagnosed were labeled as potentially lethal, serious and significant. The data persons with schizophrenia in reducing internalized stigma and in was statistically compared by unpaired samples test and the effi- boasting their self-esteem. The aim of the present study is to ex- cacy of the intervention on type of errors was margined by chi plore the predictive relationship of social support with inter- square test. nalized stigma and self-esteem among diagnosed persons with Results: 8.5 medication errors per 1000 prescription orders schizophrenia. were identified (1.5 medication error per 50 patient per day), Methods: A purposive sample of 52 diagnosed persons with where-as 18 potentially lethal errors were identified within the schizophrenia, age ranges 18 to 55 years who belong to different span of 8 months. Potential cardiopulmonary arrest and anaphy- socioeconomic status was taken from different psychiatric hospi- lactic risk was significantly assessed (po0.01) in ICU. Overdose tals of Karachi. Demographic Information sheet, Multi-Dimen- and under dose of antibiotics accounted to 76% errors. The fre- sional Scale of Perceived Social Support, Internalized Stigma of quency of errors was comparatively non significant in prescription Mental Illness Scale (ISMI) and Rosenberg Self Esteem Scale were orders by senior attendees (po0.05). administered individually. Conclusions: Effective control of antibiotic medication errors is Results: Findings revealed social support predicts internalized observed with the integrated role of pharmacist in the prescrip- stigma and self-esteem in persons with schizophrenia (po0.000). tion-dispensing-administration sequence. Conclusions: There is an immense need to develop awareness SIMULTANEOUS DETERMINATION OF SIX ANTIHISTAMINE regarding social support group and to generate strategies to re- ANTIALLERGIC DRUGS IN PHARMACEUTICAL FORMULATIONS, duce the effect of internalized stigma in person with severe mental HUMAN SERUM AND PHARMACOKINETICS APPLICATION illness. Nawab Sher1*, Farhan Ahmed Siddiqui2, Najmul Hasan3and EFFICACY AND SAFETY OF METFORMIN IN GESTATIONAL Nighat Shafi2 DIABETES MELLITUS 1Department of Chemistry, Faculty of Science, University of Kar- Huma Muzaffar1, Shama Chaudhary1and Rubina Hussain1 achi, Pakistan,2Faculty of Pharmacy, Federal Urdu University of Arts, 1Ziauddin University, Karachi, Pakistan. Science and Technology, Karachi, Pakistan,3Department of Micro- Objective: To assess the efficacy and safety of Metformin for biology, Faculty of Science, University of Karachi, Pakistan. the management of gestational diabetes Mellitus (24 wks and This article describes a new, accurate and highly specific high above) as compared to Diet. performance liquid chromatographic method with UV detection Methods: This Quasi experimental study is being conducted on (HPLC-UV) for the simultaneous determination of cetirizine HCl, 600 females at Ziauddin University Hospital. All pregnant women chlorphenamine maleate, loratadine, domperidone, buclizine and receiving prenatal care fulfilling the inclusive criteria will be meclizine in pharmaceutical dosage form and human serum, in- screened using a 50g glucose challenge test (GCT). After the result volving pyridoxine as internal standard. The mobile phase consists if it is positive, women will be advised for oral glucose tolerance of heptane sulphonic acid salt buffer and acetonitrile, drawn at a test. After diagnosis of gestational diabetes Mellitus patients will flow rate of 1.0 mL/min using a symmetry C18 column with UV be advised for diet control or oral hypoglycemic drug metformin detection at 230 nm. The intraday and inter-day precision mea- starting from 500mg twice a day. Patient will be followed fort- surements showed coefficients of variation always less than one. nightly with sugar record for adjustment of treatment. Patient will The calibration curve was tested in the range of 10–2150 ng/mL be followed till delivery for any maternal or perinatal complica- and the correlation coefficient of 40.9990 in all cases was ob- tions. Patients with systemic diseases are excluded from the study. tained. The averages of the absolute and relative recoveries were Results: Result of this study are awaited as this is an ongoing found to be in the range of 98 to 102%. Up to six drugs were se- study. parated in the same chromatogram with good resolution. The Conclusions: The main purpose of the treatment is to prevent proposed HPLC method has reasonable applications in pharma- fetal hyperinsulinemia and fetal macrosomia by reducing maternal ceutical tablet dosage form and pharmacokinetics studies. glucose level, decrease risk of appearance of diabetes in sub- SIMULATION BASED CARDIOPULMONARY AUSCULTATION: A sequent pregnancies and in future without pregnancy. Metformin TOOL TO AID MEDICAL STUDENTS OF ZIAUDDIN UNIVERSITY is an alternative to insulin and is effective in the treatment of Iqrah Iqbal1, Kulsoom Farooqi1and Fatima Z Mithani1 gestational diabetes. 1Ziauddin University, Karachi, Pakistan. IMPACT OF PHARMACIST INTERVENTION ON ANTIBIOTIC Introduction and Objectives: Use of simulation and state of art PRESCRIPTION ORDERS IN PEDIATRIC SETTING mannequin and models in clinical skills lab is now becoming a Uzma Shahid1, Mirza Tasawer Baig2, Ali Akbar Sial2and Zahida common practice which is offered to 3rd year medical students Memon1 during their clinical rotations. We seek to determine the efficacy of 1Faculty of Medicine, Ziauddin University, Karichi, cardiopulmonary auscultatory skills in 3rd year medical students Pakistan,2Faculty of Pharmacy, Ziauddin University, Karachi, Pakistan. of Ziauddin University who have been trained by simulation based Introduction and Objective: The prospective study was de- auscultatory device. signed to formulate and test the efficacy and impact of trial in- Methods: Students who participated in this study (n=40) were terventions by pharmacist on potential harm and incidence rate of from 3rd year MBBS program. They were assigned in 2 equal errant antibiotic medications dispensed in children admitted to a groups. Group A took a lecture on the detection and knowledge of public sector hospital in Karachi. various cardiopulmonary sounds whereas Group B was tutored on Methods: An eight months study was conducted in a teaching a simulation based device after they had taken their lecture. After hospital comprising of 850 beds with 65 beds strength in three respective training sessions, each group was tested with the help pediatric units I, II and III inclusive of neonatal ICU. Orders written of a questionnaire. This questionnaire consisted of identification by physician groups in all patient admission categories were and knowledge based open ended and close ended questions. Data N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 217 was entered into data analysis software S.P.S.S ver 15, Data was Deficiency of Insulin causs hyperglycemia and hyperlipidemia. analyzed and mean, standard deviation was calculated along with This leads to multiple disorders like atherosclerosis, CVAs, IHD, Charts, graphs were generated to represent results. Renal failure, NAFLD, glaucoma, cataracts, gangrenes of limbs, Results: Diagnostic accuracy and correct identification of car- acute emergencies like DKA (diabetic ketoacidosis) and HONKC diopulmonary sounds was better in the group that had been as- (hyperosmolar non ketotic coma) etc. NAFLD (Non Alcoholic Fatty sisted with a simulation based device. Mean score of assessment of Liver Disease) was first described by Ludwig in 1980 and is on rise, knowledge and skills of cardiopulmonary sounds were found making it the most common cause of CLD. Commonly called fatty significant in group B (mean = 13.5±1.73) compared to group A liver diseases is a major problem arising in diabetic population, (mean = 6.9±2.64). which has been neglected off until now. The prevalence rises to Conclusions: According to our study at Ziauddin medical uni- 60.8% in diabetics in Pakistan. 40% cases of NAFLD can progress to versity, it is suggested that simulation based learning is a sub- NASH (nonalcoholic steatohepatitis) of whom 32%-37% can pro- sidiary tool in preserving knowledge and skills for cardio- gress to advanced fibrosis. This prevalence rises to 35% to 75% in pulmonary auscultation. obese and diabetics. Making it most common cause of cryptogenic WHOLE FRUIT BETTER THAN FRUIT JUICES IN MAINTAINING cirrhosis, which can lead to hepatocellular carcinoma. NAFLD is BLOOD SUGAR LEVELS usually an incidental finding when an abdominal scan is done for Somia Gul1, Humera Khatoon1, Haneen Rashid1and Nameera some other suspected pathology. Ahmed1 Methods: FBS, Fasting LIPID profile andHbA1C was done. Ul- 1Faculty of Pharmacy, Jinnah University for Women, Karachi, trasound was done to see fatty infiltration of liver. Pakistan. Conclusion: Ultrasound liver of type II diabetic patients Introduction and Objectives: Type 2 diabetes mellitus (T2DM), showing non-alcohol fatty liver changes. non-insulin dependent diabetes mellitus, is the most rapidly IDENTIFYING ERRORS IN PRESCRIPTION WRITING growing disease, estimated to affect 439million adults by 2030. S H Afsheen Nazar1, Nasim Karim1and Riffat Farooqui1 The rate is faster in developing countries than developed countries 1Department of Pharmacology and Therapeutics, Bahria University which in turn results in higher healthcare expenditure. Thus it Medical and Dental College, Karachi, Pakistan. becomes a public health priority to prevent the disease and con- Introduction and Objective: The errors of prescribing are the trol over its rapid growth. Among the known risk factors that commonest form of avoidable medication errors and are con- contribute to T2DM are dietary factors, which promote early dia- sidered to be the most important target for improvement. Medi- betes in healthy individuals and increased mortality and morbidity cation errors are common in general practice. Both errors in the in type 2 diabetic patients. To evaluate the hypothesis that fruits act of writing (prescription errors) and prescribing faults due to provide more protection against T2DM than fruit juice, we per- erroneous medical decisions can result in harm to patients. Our formed experiment by giving fruits to one group and fruit juice to objective is to identify the errors in the prescriptions of general another group of healthy volunteers and monitor fasting blood practitioners (GPx) from different parts of Karachi. sugar and random blood sugar. Methods: A descriptive pilot study was conducted from 1st Results: The results of current study suggested that whole January to 30th February 2014. A total of 100 prescriptions were grapefruit and orange fruit are very effective in maintaining ran- collected from 4 general practitioners’clinics (east, west, central dom blood sugar levels in comparison to juices. and south districts) of Karachi. Verbal consent was taken few days Results also suggested that dietary fibers are not only effective prior to random collection. All prescriptions were analysed for in regulating blood sugar, but also showed insignificant change in errors in superscription, inscription, subscription, transcription, systolic and diastolic blood pressure and also in pulse rate. signatures and refill information. Moreover, whole grapefruit has been found more effective in Results: A total of 373 errors were identified in 100 prescrip- maintaining pulse rate in comparison to fibers obtained from ap- tions. 203 errors in superscription, 6 in subscription, 34 in tran- ple fruit and orange fruit. scription, 1 in prescribers’ signatures and 62 in refill information. Conclusions: Based on above findings, it has been concluded Conclusion: Errors in prescription writing are found to be that patients with type 2 DM could be maintained on dietary fibers common in the prescriptions of general practitioners. Measures not only for better glycemic control, but also for controlling dia- should be taken to refresh the prescription writing skills of general betes related complication such as high blood pressure. This study practitioners through CMEs and workshops. could further be extended to assess the effects of dietary fibers ASSESSMENT OF STRESSORS AMONG UNDERGRADUATE (especially obtained from grapefruit) in controlling other diabetes DENTAL STUDENTS OF KARACHI, PAKISTAN related complications, e.g. hypercholesterolemia, diabetic ne- Safia Anwar1and Saira Hamid1 phropathy, in diabetic patients. 1Bahria University Medical and Dental College, Karachi, Pakistan. . Introduction and Objectives: Dental schools are known to NAFLD IN DIABETIC PATIENTS TREATED ON ORAL have highly stressful learning environments. Dentistry involves an HYPOGLYCEMIC. acquisition of required academic, clinical and interpersonal skills Dunesh Kumar1, Zahida Memon1and Samia Perwaiz1 during the course of learning. Identifying sources of stress re- 1Department of Pharmacology, Ziauddin University, Karachi, presents the crucial first step towards advocating policy changes Pakistan. and enhance student’s stress coping skills. This study was con- Introduction and Objectives: Determination of NAFLD (non- ducted to assess the problems related to stress encountered by alcoholic fatty liver disease) in diabetic patients on oral hypogly- dental students of Karachi. The aims are: a) To identify the com- cemic therapy. Diabetes is one of the most common non-com- mon stressors among undergraduate dental students of Karachi. b) municable disorder throughout the world. It has enormous com- To investigate whether specific stressor is related to year of study plications, which indirectly multiply the economic and medical or gender. c) To evaluate the factors which influence student burden. According to WHO, 9% population of the world suffers perception that might allow programmatic changes. from Diabetes who are above 18 years age. Prevalence of diabetes Methods: Type of Study: Cross Sectional Survey; Sample Size: in Pakistan is about 10% according to Akhtar. This poses a huge 330 undergraduate dental students of Karachi; Study population: burden on a country like Pakistan, which already has very limited second, third and final year BDS; Study setting: Five dental schools resources in field of health. of Karachi; Modified form of Dental Environment Stress 218 N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 questionnaire was administered. history of tobacco usage, chewable or smoke products and details Results: A total of 500 Questionnaire were distributed out of of chewing habits (using since, packs/day, duration of exposure which only 330 responded. Of the respondent, 70% were women etc.). The study was carried out in Ziauddin. and 30% were men. The most stressful variable among all was Results: Majority of the subjects were males (80%). Out of the ‘Overloaded feeling due to vast syllabus’. Third and final year 71 Oral Squamous Cell Carcinoma patients majority 42 (59.15%) students were reported to have higher-level stress than second were multiple users that consumed more than one tobacco pro- year students. Females were more affected by the same stressor duct. Whereas, 9 (12.67%) consumed gutka, 9 (12.67%) pan, 5 (7%) than male students. betel nut, 4 (5.6%) niswar and 2 (2.8%) cigarette. Conclusions: The conclusion of study was drawn from vari- Conclusions: Frequency of OSCC was found higher in users of ables with significant values o0.005. Stress due to accommoda- multiple chewable tobacco formulations which makes consump- tion problem was higher in students of second year while clinical tion of multiple tobacco formulations one of the major etiological based problem caused greater level of stress among students of factors in the development of OSCC. third and final year. The variables relating to academics (in- MOLECULAR JOURNEY OF VITAMIN D, VDR AND VDRGP AND adequate break timings, inadequate time for assigned work and ITS ASSOCIATION WITH BREAST CANCER overloaded due to vast syllabus) caused stress in all three years Samiuddin Ahmed1, Saiyed Abdullah Abubakar1and Bushra equally. Stress showed an upward trend from the first to the final Wasim1 year with a jump from the second to the third years. 1Ziauddin University, Karachi, Pakistan. FREQUENCY OF DEPRESSION IN HOUSE WIVES AND ASSO- Vitamin D plays an essential role in regulating the levels of CIATED RISK FACTORS calcium and phosphate in the body. Vitamin D is not active as a Sarah Baber1, Sana Ilyas1and M Ammar Hassan1 hormone - instead, a few chemical changes must be made. These 1Ziauddin University, Karachi, Pakistan. are performed by enzymes that add hydroxyl groups. Vitamin D Introduction and Objectives: In any population, women are receptor is a nuclear hormone receptor Transcriptional regulator in more prone to depression. In our middle class community it is still response to 1,25,dihydroxyvita-min D 3. considered a taboo. Housewives usually emphasize on somatic VDR protein is at the centre of the Vitamin D endocrine system issues rather than emotional signs and symptoms which causes with a negative feedback which regulates serum calcium and Vi- detrimental effects on their health. Timely identification of risk tamin D, if not maintained will affect large number of organs in- factors and counseling of women regarding their mental health is cluding breast. Data is available in positive genetic association of of utmost importance. We aim to measure frequency and identify certain diseases which suggest functional consequences of VDR major risk factors of depression in housewives belonging to mid- gene polymorphism. Vitamin D plays a role in breast cancer and dle class community in Karachi. exerts its effect through VDR gene polymorphism. There are four Methods: Cross sectional study conducted on housewives be- polymorphic variants of VDR (Folk 1, Bsm1, Taq 1 and Appa1). tween 25-50 years of age. Through convenience sampling tech- Hormone Reception-Vitamin D hormone binds to receptors in its nique 247 questionnaires with 17 questions were administered, target cells, controlling the synthesis of many different proteins followed by the patient’s health questionnaire 9 (PHQ-9). Data was involved in calcium transport and utilization. The receptor is analyzed by SPSS v20. Chi-test was calculated for categorical data composed of two domains: a domain that binds to the hormone and p-value o0.05 was considered significant. and a domain that binds to DNA. It pairs up with a similar protein, Results: Out of all the respondents, 131 females (53%) turned 9-cis retinoic acid receptor (RXR), and together they bind to the out to be depressed and 116 (57%) not depressed. PHQ-9 classifi- DNA, activating synthesis in some cases and repressing it in others. cation of the levels of depression in the participants showed 100 There is a decrease in VDR expression in breast cancer cells com- females mildly depressed, 77 females moderately depressed, 35 pared to normal breast cells and this decrease could be due to females moderately-severely depressed, 16 females severely de- gene polymorphism and other factors also Alterations in VDR ex- pressed and 19 females not depressed. pression and activity could lead degranulation of vitamin D up- Conclusions: Most prominent risk factors for depression were take, metabolism and serum levels of biologically active Vitamin D. of having to take care of an old or an ill person at home and living POTENTIAL OF BLACK MULBERRY: FROM FOOD OF CHOICE in a joint family structure. Although the study targeted middle TO REMEDY FOR BOWEL DYSMOTILITY class community, the analysis showed no associations with the Malik H Mehmood1*, Anam Akhlaq1, Zohaib Ashraf1, Sadia Syed1, family income. Samira A Bawany1, Abdul Rehman1, Anwarul-Hassan Gilani1,2 FREQUENCY AND TYPE OF CHEWABLE TOBACCO COMMONLY 1Natural Products Research Division, Department of Biological and USED BY ORAL SQUAMOUS CELL CARCINOMA PATIENTS Biomedical Sciences, Aga Khan University, Karachi, Pakistan,2Pakistan Fauzia Perveen1and Saeeda Baig1 Council for Science and Technology, Islamabad, Pakistan. 1Department of Biochemistry, Ziauddin University, Karachi, Introduction and Objectives: Morus nigra Linn. (black mul- Pakistan. berry) is used in gastrointestinal ailments. This study demon- Introduction: South East Asia is threatened by the risk of Oral strates gut modulatory proper-ties of M. nigra. Squamous Cell Carcinoma. The popularity of chewable tobacco Methods: The prokinetic, laxative and antidiarrheal activities of plays a pivotal role in making it the second leading malignancy in M. nigra were assessed in mice, while isolated rabbit jejunum and Pakistan. Prolonged use of these products can lead to different oral guinea-pig ileum were used to explore insight into mechanism(s). precancerous conditions, GI cancer, ulceration and many other Results: At 30 and 70 mg/kg, the crude extract of M. nigra (Mn. diseases. The objective of the study was to find out the frequency Cr) exhibited atropine-sensitive prokinetic and laxative effects, and type of chewable tobacco, which has been the cause of Oral similar to carbachol (CCh). While at higher doses (100-500 mg/kg), Squamous Cell Carcinoma in these patients. Mn.Cr offered protection against castor oil-induced diarrhea. Methods: The study was descriptive. Seventy one patients who In rabbit jejunum, Mn.Cr and its chloroform fraction inhibited had Oral Squamous Cell Carcinoma and a history of tobacco use CCh-induced contractions more potently compared to high K+ (80 were selected from cancer OPD. Subjects addicted to stuff other mM). Conversely, petroleum fraction was more potent against high than smoking and chewable tobacco were excluded. A ques- K+-induced contractions. At 0.01 mg/mL, Mn.Cr caused a parallel tionnaire was used to collect demographic characteristics, oral shift in acetylcholine (ACh) concentration-response curves (CRCs) hygiene, medical history, family history of cancer, and detailed followed by a non-parallel shift at 0.03 mg/mL, similar to N.A. Siddiqui, A. Shahzad / New Horizons in Translational Medicine 3 (2016) 175–219 219 dicyclomine. At further tested concentrations, Mn.Cr (0.1 and 0.3 Methods: The study was preformed to evaluate the anti- mg/mL) and petroleum fraction suppressed Ca2+ CRCs, similar to microbial activity of honey and turmeric, purchased from local verapamil. In guinea-pig ileum, Mn.Cr, its aqueous and ethyl market. Different concentrations of honey and turmeric were acetate fractions exhibited atropine-sensitive gut stimulant activ- prepared in distilled water including 100%, 75% and 50%. Its effect ity along with additional uncharacterized excitatory response in was observed on isolates obtained from clinical samples, which aqueous fraction only. includes Salmonella typhi, Shigella dysentrey, S.aureus, Proteus Conclusions: These results suggest that black mulberry pos- mirabilis, E.coli, Pseudomonas aerignosa, Klebsiella oxytoca, Ba- sesses prokinetic, laxative and antidiarrheal effects, putatively cillus subtilis, Enterococcus and Micrococcus leutus. The study was mediated through cholinomimetic and anti-muscarinic plus Ca2 carried by disc diffusion method. All experiments were performed +-antagonist mechanisms, respectively. in tricplicate. Zone of inhibition was measured in millimeter and ANTIBACTERIAL ACTIVITY OF HONEY AND TURMERIC results were given in percentage. AGAINST GIT AND RESPIRATORY Results: Honey extract show highest antibacterial activity as PATHOGENS compared to turmeric extract. Highest concentration (100%) of Hira Jalal1and Aliya Hayat1 1Department of Microbiology, Jinnah University for Women, Kar- honey and turmeric was inhibitory to all strains among which achi, Pakistan. Enterococcus gave 75% inhibition while other strains were in- Introduction and Objectives: Because of emergence of re- hibited up to 50%. It was found that 75% concentration of these sistant pathogens the antibiotic activity is diminished. This poses a compounds gave intermediate zone of inhibition while 50% con- serious threat to public health. Therefore alternative antimicrobial centration was least inhibitory. techniques are needed. Thus this situation has led to reevaluation Conclusions: Honey and turmeric have potential to use as of the therapeutic use of ancient remedies such as plants and plant therapeutic agents either alone or in combination against various based products including honey and turmeric. infections.