DOCSLIB.ORG

Induction of HL-60 Leukemia Cell Differentiation by the Novel Antifolate 5,10- Dideazatetrahydrofolic Acid1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Induction of HL-60 Leukemia Cell Differentiation by the Novel Antifolate 5,10- Dideazatetrahydrofolic Acid1 (CANCER RESEARCH 49. 4824-4828. September I. 1989] Induction of HL-60 Leukemia Cell Differentiation by the Novel Antifolate 5,10- Dideazatetrahydrofolic Acid1 John A. Sokoloski, G. Peter Beardsley, and Alan C. Sartorelli2 Departments of Pharmacology and Pediatrics and Developmental Therapeutics Program, Comprehensive Cancer Center, Yale University School of Medicine, New Haven, Connecticut 06510 ABSTRACT as an inhibitor of folate metabolism which acted at an enzymatic level other than dihydrofolate reducíase,the site of action of The novel tetrahydrofolate, 5,10-dideazatetrahydrofolic acid most classical antifolates, including aminopterin and metho- (DDATHF), was designed as an inhibitor of folate metabolism at a site other than dihydrofolate reducíase.DDATHF has been shown to inhibit trexate (15, 16). DDATHF closely resembles tetrahydrofolate, glycinamide ribonucleotide transformylase, a folate-requiring enzyme that thereby retaining the structural properties necessary for mem catalyzes the first of two one-carbon transfer reactions in the de novo brane transport and for it to serve as a substrate for folylpoly- purine nucleotide biosynthetic pathway. Incubation of HL-60 promyelo- glutamate synthetase (16). The presence of a fully reduced ring cytic leukemia cells with 5 x 10~* to IO"5 M DDATHF resulted in a and the 2-amino-4-oxo-substituents rather than the 2,4-dia- marked inhibition of growth after 48 h, with a complete cessation of mino-substitution pattern found in classical antifolates presum cellular replication by day 4. Cell cycle analyses of DDATHF-treated ably are responsible for the lack of inhibition of dihydrofolate HL-60 cells demonstrated an initial block in early S phase by day 3 reducíaseby DDATHF. followed by an accumulation of cells in the G, and <. + M phases of the Reduced folates particípalein a variely of one-carbon transfer cell cycle. Inhibition of growth was accompanied by a concentration- reaclions important for cell growlh, such as Ihe production of dependent increase in the percentage of mature myeloid cells that ex pressed nitroblue tetrazolium positivity, and a small increase in nonspe melhionine from homocysteine, the interconversion of serine cific esterase activity. Induction of differentiation and inhibition of growth and glycine, and the calabolism of several amino acids, in by DDATHF were completely prevented by hypoxanthine and 5(4)- addition to Iheir role in Ihe de novo production of purine amino-4(5)-imidazole carboxamide, suggesting that depletion of intracel- nucleotides and Ihymidylale (17). The presence of carbon aloms lular purine nucleotide pools has an important role in the biological in place of nilrogens al posilions 5 and 10 necessarily prevenís effects of this inhibitor. This possibility was confirmed by the finding DDATHF from parlicipaling as a cofactor in Ihese reaclions, that DDATHF caused a pronounced reduction in intracellular GTP and and also provides for increased chemical slabilily. ATP levels within 2 h, with maximum decreases being observed by 24 h, Previous studies with DDATHF have indicated that the a time interval which preceded the inhibition of cellular proliferation by parenl compound or a polyglutamyl derivative is a potent this agent. Pyrimidine nucleoside triphosphate levels were markedly inhibitor of de novo purine nucleolide biosynlhesis (16, 18). increased under these conditions. The findings indicate the importance of purine nucleotides to both the inhibition of growth and the induction of Furthermore, evidence has been presenled lo suggesl lhal GAR differentiation of HL-60 leukemia cells by DDATHF. Iransformylase, Ihe first of Iwo folale-requiring enzymes lhal calalyze one-carbon Iransfer reaclions in Ihe de novo purine nucleolide biosynlhelic palhway is the primary cellular target INTRODUCTION of this agent (18). Since the cyloloxicily of DDATHF in cul- The HL-60 promyelocytic leukemia has been used extensively lured cells appears lo be dependeni upon Ihe depletion of as a model of hematopoietic development (1). HL-60 cells are inlracellular purine nucleolide levels, it was of interesl lo sludy multipotent and have been shown to be capable of differentiat Ihe effecls of DDATHF on Ihe growlh and differenlialion of ing in vitro to cells with characteristics of mature granulocytes HL-60 promyelocylic leukemia cells. (2, 3) or monocytes (4, 5) by a variety of different inducing The presenl sludy demonslrales lhal DDATHF is a polenl agents including several antitumor agents (6-9). The differen inducer of Ihe maluration of HL-60 leukemia cells. Further tiation of HL-60 cells to eosinophils and basophils has also more, Ihe findings indicale lhal Ihe induclion of differentiation been reported (10, 11). Among the initiators of maturation are by DDATHF is closely associated with Ihe inhibilion of de novo the novel C-nucleoside, tiazofurin, and related inhibitors of purine nucleotide biosynthesis, presumably al Ihe reaclion cal- IMP dehydrogenase (12, 13). The induction of the differentia alyzed by GAR Iransformylase. tion of HL-60 cells by these agents, which diminishes intracel lular GTP pools, suggests that GTP depletion may be involved MATERIALS AND METHODS in modifying intracellular signals which increase the probability of a cell entering a differentiation pathway. The de novo inhib Cell Culture. HL-60 promyelocytic leukemia cells were originally itor of purine nucleotide biosynthesis, 6-methylmercaptopurine supplied by Dr. Robert C. Gallo of the National Cancer Institute, ribonucleoside, which produces depletion of both GTP and Bethesda, MD. The HL-60 cells used in these experiments were ex ATP, is also an effective inducer of HL-60 maturation (14). panded from stocks frozen in liquid nitrogen, and consisted of cells The novel tetrahydrofolate, DDATHF' (Fig. 1), was designed between passages 25 and 65. Cell stocks were routinely checked for Mycoplasma contamination by the gene probe method (Gen-Probe, Received 11/8/88; revised 5/11/89: accepted 6/7/89. Inc., San Diego, CA). The costs of publication of this article were defrayed in part by the payment Cells were routinely passaged in RPMI 1640 medium (GIBCO, of page charges. This article must therefore be hereby marked advertisement in Grand Island, NY) supplemented with 15% heat-inactivated (50°Cfor accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 30 min) fetal calf serum (GIBCO). Cultures were maintained at 37°C 1This research was supported in part by USPHS Grants CA-02817 and CA- 42300 from the National Cancer Institute. in a humidified atmosphere containing 5% COs in air. Experiments - To whom requests for reprints should be addressed, at Department of were performed by resuspending cultures of HL-60 cells at a density of Pharmacology. N'aie University School of Medicine, P. O. Box 3333. New Haven. 5 x IO4 cells/ml in the presence of DDATHF for 7 days at 37°C. CT 06510-8066. Solutions of DDATHF were prepared in 0.1 N NaOH and adjusted to 'The abbreviations used are: DDATHF. 5.10-didea/atetrahydrofolic acid; IO"3 M in PBS using an £:7,nm (0.1 N NaOH) value of 9.15 x 10"* PBS, phosphate-buffered saline: NBT. nitroblue tetra/olium: AICA. 5(4)-amino- 4(5)-imida/ole carboxamide: GAR, glycinamide ribonucleotide. M~' cm~' (18). Cell numbers were determined daily with a Coulter 4824 Downloaded from cancerres.aacrjournals.org on October 4, 2021. © 1989 American Association for Cancer Research. DDATHF INDUCTION OF DIFFERENTIATION COOH | Fig. 1. Chemical structure of DDATHF. particle counter equipped with a channelizer (Coulter Electronics, Hialeah, FL). Viable cells were ascertained by using a 0.1 % try pun blue solution and a hemocytometer. Smears were prepared with a Shandon- Southern Cytospin (Johns Scientific, Toronto, Ontario, Canada) and were stained with May-Grunwald-Giemsa. 0 48 96 144 Assessment of Differentiation. The capacity of HL-60 cells exposed Time (h) to DDATHF to undergo functional maturation was determined by NBT dye reduction (19). Approximately 1 x IO6cells were collected by Fig. 2. Effects of DDATHF on the proliferation of HL-60 leukemia cells. Five x 10* cells/ml were incubated with various concentrations of DDATHF for 7 centrifugation and resuspended in 1.0 ml of RPMI 1640 medium days. Cell numbers were measured daily with a Model ZBI Coulter particle containing 0.1% NBT (Sigma Chemical Co., St. Louis, MO) and 1.0 counter. Treatment: none (x), 0.01 >IM(D), 0.1 >iM(T), 1.0 >IM(A), and 10 >IM /ig/ml of 12-0-tetradecanoylphorbol-13-acetate (Sigma) in ethanol. The (•)DDATHF. cell suspension was incubated for 30 min at 37°C,and the percentage of cells containing blue-black formazan granules, indicative of a 12-0- B.S teiradecanoylphorbol-13-acctale-stimulated respiratory burst, was de G, -36.7 •¿37.a termined microscopically. Nonspecific-esterase (a-naphthylbutyrate es S -442 -40.1 S •¿40.0 G,»M- 19.1 „¿.UC.¡G,az*M'23.l GjtM'22.7y terase) activity was monitored as described by Yam et al. (20). Flow Cytometry. Flow cytometry was performed by using a FACS IV fluorescence-activated cell sorter (Becton Dickinson, Mountain View, CA). Cells were washed twice in ice-cold PBS, fixed in 70% ethanol, and stored at 4°Cfor up to 2 weeks prior to analysis. Histo grams of the relative DNA content of induced cells were obtained from » w* D. 30,000 cells stained with SO/¿I/mlofpropidium iodide after treatment O G, »14.8 ' 39.9 with 1 mg/ml of RNase for 30 min at 37°C.The percentage of cells in S -62.1 •¿40.6 G,»M'23.I the G,, S, and G2 + M compartments of the cell cycle was determined by the method of Jett (21). Analysis of Ribonucleotide Pools. HL-60 leukemia cells (1 to 2 x 10' cells/ml) were suspended in fresh RPMI 1640 medium containing 10 '' M DDATHF.
Load more

Recommended publications
  • 35 Disorders of Purine and Pyrimidine Metabolism
    35 Disorders of Purine and Pyrimidine Metabolism Georges van den Berghe, M.- Françoise Vincent, Sandrine Marie 35.1 Inborn Errors of Purine Metabolism – 435 35.1.1 Phosphoribosyl Pyrophosphate Synthetase Superactivity – 435 35.1.2 Adenylosuccinase Deficiency – 436 35.1.3 AICA-Ribosiduria – 437 35.1.4 Muscle AMP Deaminase Deficiency – 437 35.1.5 Adenosine Deaminase Deficiency – 438 35.1.6 Adenosine Deaminase Superactivity – 439 35.1.7 Purine Nucleoside Phosphorylase Deficiency – 440 35.1.8 Xanthine Oxidase Deficiency – 440 35.1.9 Hypoxanthine-Guanine Phosphoribosyltransferase Deficiency – 441 35.1.10 Adenine Phosphoribosyltransferase Deficiency – 442 35.1.11 Deoxyguanosine Kinase Deficiency – 442 35.2 Inborn Errors of Pyrimidine Metabolism – 445 35.2.1 UMP Synthase Deficiency (Hereditary Orotic Aciduria) – 445 35.2.2 Dihydropyrimidine Dehydrogenase Deficiency – 445 35.2.3 Dihydropyrimidinase Deficiency – 446 35.2.4 Ureidopropionase Deficiency – 446 35.2.5 Pyrimidine 5’-Nucleotidase Deficiency – 446 35.2.6 Cytosolic 5’-Nucleotidase Superactivity – 447 35.2.7 Thymidine Phosphorylase Deficiency – 447 35.2.8 Thymidine Kinase Deficiency – 447 References – 447 434 Chapter 35 · Disorders of Purine and Pyrimidine Metabolism Purine Metabolism Purine nucleotides are essential cellular constituents 4 The catabolic pathway starts from GMP, IMP and which intervene in energy transfer, metabolic regula- AMP, and produces uric acid, a poorly soluble tion, and synthesis of DNA and RNA. Purine metabo- compound, which tends to crystallize once its lism can be divided into three pathways: plasma concentration surpasses 6.5–7 mg/dl (0.38– 4 The biosynthetic pathway, often termed de novo, 0.47 mmol/l). starts with the formation of phosphoribosyl pyro- 4 The salvage pathway utilizes the purine bases, gua- phosphate (PRPP) and leads to the synthesis of nine, hypoxanthine and adenine, which are pro- inosine monophosphate (IMP).
    [Show full text]
  • Forms of Hypoxanthine Or, Through Interconversions, Guanine Or Adenine
    826 GENETICS: GOTS AND GOLLUB PROC. N. A. S. Summary.-The ribonucleic acids of isolated thymus nuclei can be separated into two distinct fractions, one of which probably represents ribonucleic acid of the nu- cleolus. Studies of the incorporation of orotic acid-6-C"4 and adenosine-8-C'4 into these RNA fractions in vitro show great differences in their metabolic activity and different susceptibilities to an inhibitor of RNA synthesis, the "nucleolar" RNA being by far the more active. It is a pleasure to acknowledge our indebtedness to Mr. Rudolf Meudt for his careful and expert technical assistance. * This research was supported in part by a grant (RG-4919 M&G) from the United States Public Health Service. I V. G. Allfrey, A. E. Mirsky, and S. Osawa, Nature, 176, 1042, 1955. 2 V. G. Allfrey, A. E. Mirsky, and S. Osawa, J. Gen. Physiol., 40, 451, 1957. 3 R. Logan and J. N. Davidson, Biochim. et Biophys. Acta, 24, 196, 1957. 4 S. Osawa, K. Takata, and Y. Hotta (in press). 6 J. M. Webb, J. Biol. Chem., 221, 635, 1956. 6 M. Bessis, in Traite de cytologie sanguine (Paris: Masson & Cie, 1954), p. 83. J. N. Davidson and R. M. S. Smellie. Biochem. J.. 52, 594, 1952. SEQUENTIAL BLOCKADE IN ADENINE BIOSYNTHESIS BY GENETIC LOSS OF AN APPARENT BIFUNCTIONAL DEACYLASE* By JOSEPH S. GOTS AND EDITH G. GOLLUB DEPARTMENT OF MICROBIOLOGY, SCHOOL OF MEDICINE, UNIVERSITY OF PENNSYLVANIA, PHILADELPHIA, PENNSYLVANIA Communicated by D. Wright Wilson, July 3, 1957 Along the biosynthetic pathway to the purines of nucleic acids, inosinic acid occurs as a pivotal point in a bifurcation which leads to adenylic acid along one branch and to guanylic acid along the other: (B) r- Adenylic acid (AMP) (A) Inosinic acid (IMP) (C) L|_ * Guanylic acid (GMP) Bacterial mutations may result in genetic impairments at three locations with respect to the pivotal inosinic acid, thus yielding auxotrophs with three broad classes of nutritional response.
    [Show full text]
  • A Fairy Chemical, Imidazole-4-Carboxamide, Is
    www.nature.com/scientificreports OPEN A Fairy Chemical, Imidazole-4- carboxamide, is Produced on a Novel Purine Metabolic Pathway in Received: 25 September 2018 Accepted: 14 May 2019 Rice Published: xx xx xxxx Hirohide Takemura1, Jae-Hoon Choi 2,3, Nobuo Matsuzaki1, Yuki Taniguchi2, Jing Wu3, Hirofumi Hirai2,3, Reiko Motohashi2,3, Tomohiro Asakawa4, Kazutada Ikeuchi5, Makoto Inai6, Toshiyuki Kan6 & Hirokazu Kawagishi1,2,3 Rings or arcs of fungus-regulated plant growth occurring on the foor of woodlands and grasslands are commonly called “fairy rings”. Fairy chemicals, 2-azahypoxanthine (AHX), imidazole-4-carboxamide (ICA), and 2-aza-8-oxohypoxanthine (AOH), are plant growth regulators involved in the phenomenon. The endogeny and biosynthetic pathways of AHX and AOH in plants have already been proven, however, those of ICA have remained unclear. We developed a high-sensitivity detection method for FCs including ICA and the endogenous ICA was detected in some plants for the frst time. The quantitative analysis of the endogenous level of ICA in rice and Arabidopsis were performed using 13C-double labeled ICA. In addition, the incorporation experiment and enzyme assay using the labeled compound into rice and partially purifed fraction of rice indicated that ICA is biosynthesized from 5-aminoimidazole-4- carboxamide (AICA), a metabolite on the purine metabolic pathway. The relationship between ICA and AHX was also discussed based on quantitative analysis and gene expression analysis. “Fairy rings” is a phenomenon in which fruiting bodies of higher fungi occur afer turfgrass grows and/or dies in the form of rings1. Tese diameters vary from a few centimeters to more than 15 meters and it is also known as a disease of turfgrass all over the world2,3.
    [Show full text]
  • The Therapeutic Targeting of Folate Receptor Alpha Positive Tumors
    Wayne State University Wayne State University Dissertations 1-1-2015 The Therapeutic Targeting Of Folate Receptor Alpha Positive Tumors Via Folate Receptor Selective Novel 5- And 6- Substituted Pyrrolo [2,3-D]pyrimidine Antifolates" Shermaine Kimberly Mitchell-Ryan Wayne State University, Follow this and additional works at: http://digitalcommons.wayne.edu/oa_dissertations Part of the Molecular Biology Commons, Oncology Commons, and the Pharmacology Commons Recommended Citation Mitchell-Ryan, Shermaine Kimberly, "The Therapeutic Targeting Of Folate Receptor Alpha Positive Tumors Via Folate Receptor Selective Novel 5- And 6- Substituted Pyrrolo [2,3-D]pyrimidine Antifolates"" (2015). Wayne State University Dissertations. Paper 1155. This Open Access Dissertation is brought to you for free and open access by DigitalCommons@WayneState. It has been accepted for inclusion in Wayne State University Dissertations by an authorized administrator of DigitalCommons@WayneState. THE THERAPEUTIC TARGETING OF FOLATE RECPTOR ALPHA POSITIVE TUMORS VIA FOLATE RECEPTOR-SELECTIVE NOVEL 5-AND 6- SUBSTITUTED PYRROLO [2,3-D] PYRIMIDINE ANTIFOLATES by SHERMAINE KIMBERLY MITCHELL-RYAN DISSERTATION Submitted to the Graduate School of Wayne State University, Detroit, Michigan In partial fulfilment of the requirements for the degree of DOCTOR OF PHILOSOPHY 2015 Major: CANCER BIOLOGY Approved By: ________________________________________ Advisor Date ________________________________________ ________________________________________ ________________________________________ ________________________________________ DEDICATION This dissertation is dedicated to Seamus Isaiah Ryan, my sun (son), my world and my very reason to press forward and claim every reward life has to offer. My sojourn through this process would not have been possible without the love, support and encouragement of my husband, Paul Seamus Ryan. Thank you for your steadfast devotion, patience and understanding.
    [Show full text]
  • Taxonomic Variations in the Gut Microbiome of Gout Patients With
    Méndez‑Salazar et al. Mol Med (2021) 27:50 https://doi.org/10.1186/s10020‑021‑00311‑5 Molecular Medicine RESEARCH ARTICLE Open Access Taxonomic variations in the gut microbiome of gout patients with and without tophi might have a functional impact on urate metabolism Eder Orlando Méndez‑Salazar1,2†, Janitzia Vázquez‑Mellado3, Carlos S. Casimiro‑Soriguer4,5, Joaquin Dopazo4,5,6,7, Cankut Çubuk8, Yessica Zamudio‑Cuevas9, Adriana Francisco‑Balderas9, Karina Martínez‑Flores9, Javier Fernández‑Torres9, Carlos Lozada‑Pérez10, Carlos Pineda11, Austreberto Sánchez‑González12, Luis H. Silveira13, Ana I. Burguete‑García14, Citlalli Orbe‑Orihuela14, Alfredo Lagunas‑Martínez14, Alonso Vazquez‑Gomez15, Alberto López‑Reyes16, Berenice Palacios‑González1* and Gabriela Angélica Martínez‑Nava9† Abstract Objective: To evaluate the taxonomic composition of the gut microbiome in gout patients with and without tophi formation, and predict bacterial functions that might have an impact on urate metabolism. Methods: Hypervariable V3–V4 regions of the bacterial 16S rRNA gene from fecal samples of gout patients with and without tophi (n 33 and n 25, respectively) were sequenced and compared to fecal samples from 53 healthy controls. We explored= predictive =functional profles using bioinformatics in order to identify diferences in taxonomy and metabolic pathways. Results: We identifed a microbiome characterized by the lowest richness and a higher abundance of Phascolarc- tobacterium, Bacteroides, Akkermansia, and Ruminococcus_gnavus_group genera in patients with gout without tophi when compared to controls. The Proteobacteria phylum and the Escherichia-Shigella genus were more abundant in patients with tophaceous gout than in controls. Fold change analysis detected nine genera enriched in healthy controls compared to gout groups (Bifdobacterium, Butyricicoccus, Oscillobacter, Ruminococcaceae_UCG_010, Lach- nospiraceae_ND2007_group, Haemophilus, Ruminococcus_1, Clostridium_sensu_stricto_1, and Ruminococcaceae_ UGC_013).
    [Show full text]
  • Exercise Pills: at the Starting Line
    Review Exercise Pills: At the Starting Line 1,2 3, Shunchang Li and Ismail Laher * Sedentary lifestyles, limited physical exercise, and prolonged inactivity Trends undoubtedly increase chronic diseases, including obesity, type 2 diabetes, The concept of ‘exercise pills’ has great and cardiovascular diseases. It is widely acknowledged that exercise induces potential for use in patients having low exercise compliance or in those for a number of physiological adaptations that have beneficial effects in the pre- whom regular exercise is not feasible. vention and treatment of these chronic metabolic diseases. Unfortunately, Our increased understanding of the molecular targets of physical exercise exercise compliance is extremely low and often not possible. The development makes it possible to design agents that of exercise science and molecular techniques has increased our understanding mimic the physiological benefits of of the molecular pathways responsive to exercise. Knowledge of these molec- exercise. ular targets has led to the development of chemical interventions that can mimic Current candidate exercise pills can be the beneficial effects of exercise without requiring actual muscle activity. This divided into three categories: pharma- review focuses on the concept of ‘exercise pills’ and how they mimic the effects cological agonists (AICAR, GW501516, GSK4716, SR9009), hor- produced by physical exercise including oxidative fiber-type transformation, mones (MOTS-c, irisin), and phyto- mitochondrial biogenesis, increased fat oxidation, angiogenesis, and improve- chemicals [resveratrol and (–)-epicatechin]. ment of exercise capacity. We also review candidate exercise pills, and contrast the beneficial effects and molecular mechanisms between physical exercise The signaling pathways of currently and exercise pills. described exercise pills are outlined.
    [Show full text]
  • Differential Effects of Post-Weaning Diet and Maternal Obesity
    nutrients Article Differential Effects of Post-Weaning Diet and Maternal Obesity on Mouse Liver and Brain Metabolomes Sofiane Safi-Stibler 1,2,3 , Etienne A. Thévenot 4 , Luc Jouneau 1,2 ,Mélanie Jouin 1,2, Alexandre Seyer 5,6,Hélène Jammes 1,2, Delphine Rousseau-Ralliard 1,2 , 1,2,7, , 1,2, , Christine Baly * y and Anne Gabory * y 1 UVSQ, INRAE, BREED, Université Paris-Saclay, 78350 Jouy-en-Josas, France; sofiane.safi[email protected] (S.S.-S.); [email protected] (L.J.); [email protected] (M.J.); [email protected] (H.J.); [email protected] (D.R.-R.) 2 Ecole Nationale Vétérinaire d’Alfort, BREED, 94700 Maisons-Alfort, France 3 Collège Doctoral, Sorbonne Université, 75005 Paris, France 4 CEA, LIST, Laboratory for Data Sciences and Decision, MetaboHUB, 91191 Gif-Sur-Yvette, France; [email protected] 5 Profilomic SA, 92100 Boulogne-Billancourt, 75008 Paris, France; [email protected] 6 MedDay Pharmaceuticals, 75008 Paris, France 7 NBO, INRAE, Université Paris-Saclay, 78350 Jouy-en-Josas, France * Correspondence: [email protected] (C.B.); [email protected] (A.G.); Tel.: +33-13465-2348 (A.G.) These authors have contributed equally to this work. y Received: 17 April 2020; Accepted: 24 May 2020; Published: 28 May 2020 Abstract: Nutritional changes during developmental windows are of particular concern in offspring metabolic disease. Questions are emerging concerning the role of maternal weight changes before conception, particularly for weight loss, in the development of diet-related disorders. Understanding the physiological pathways affected by the maternal trajectories in the offspring is therefore essential, but a broad overview is still lacking.
    [Show full text]
  • Caracterización Proteómica De La Interacción Y La Inserción De Proteínas De Membrana
    TESIS DOCTORAL Valencia, Octubre 2019 Doctorado en Biomedicina y Biotecnología Caracterización proteómica de la interacción y la inserción de proteínas de membrana Dirigida por: Dr. Manuel Mateo Sánchez del Pino Dr. Luis Martínez Gil Doctorado en Biomedicina y Biotecnología Departamento de Bioquímica y Biología Molecular Facultad de Biología, Campus Burjassot Valencia, Octubre 2019 Caracterización proteómica de la interacción y la inserción de proteínas de membrana Memoria realizada por Dª Natalia Mara Vera Velasco para optar al Grado de Doctor por la Universitat de València Dirigida por: Dr. Manuel Mateo Sánchez del Pino Dr. Luis Martínez Gil OBJETIVOS • Objetivo 1 Estudio del interactoma asociado a la inserción de proteínas de membrana. • Objetivo 2 Estudio de las proteínas humanas incluidas en el virión del virus Nipah. 1 2 Objetivo 1 Estudio del interactoma asociado a la inserción de proteínas de membrana. 3 4 CONTENIDOS OBJETIVO 1 OBJETIVOS ................................................................................................................................ 1 ABREVIATURAS ....................................................................................................................... 7 INTRODUCCIÓN ....................................................................................................................... 9 I1.1 Componentes lipídicos ..................................................................................................... 12 I1.2 Componentes proteicos ...................................................................................................
    [Show full text]
  • Subtractive Proteomics to Identify Novel Drug Targets and Reverse
    www.nature.com/scientificreports OPEN Subtractive proteomics to identify novel drug targets and reverse vaccinology for the development Received: 1 March 2018 Accepted: 17 May 2018 of chimeric vaccine against Published: xx xx xxxx Acinetobacter baumannii Vandana Solanki & Vishvanath Tiwari The emergence of drug-resistant Acinetobacter baumannii is the global health problem associated with high mortality and morbidity. Therefore it is high time to fnd a suitable therapeutics for this pathogen. In the present study, subtractive proteomics along with reverse vaccinology approaches were used to predict suitable therapeutics against A. baumannii. Using subtractive proteomics, we have identifed promiscuous antigenic membrane proteins that contain the virulence factors, resistance factors and essentiality factor for this pathogenic bacteria. Selected promiscuous targeted membrane proteins were used for the design of chimeric-subunit vaccine with the help of reverse vaccinology. Available best tools and servers were used for the identifcation of MHC class I, II and B cell epitopes. All selected epitopes were further shortlisted computationally to know their immunogenicity, antigenicity, allergenicity, conservancy and toxicity potentials. Immunogenic predicted promiscuous peptides used for the development of chimeric subunit vaccine with immune-modulating adjuvants, linkers, and PADRE (Pan HLA-DR epitopes) amino acid sequence. Designed vaccine construct V4 also interact with the MHC, and TLR4/MD2 complex as confrm by docking and molecular dynamics simulation studies. Therefore designed vaccine construct V4 can be developed to control the host-pathogen interaction or infection caused by A. baumannii. Acinetobacter baumannii, an ESKAPE pathogen, has gained the attention of medical fraternity worldwide due to its nosocomial infection in hospital setup mainly ICUs and emergence of multi-drug resistance mechanism in it1–6.
    [Show full text]
  • Pyrrolo[2,3-D]Pyrimidine Classical
    Duquesne University Duquesne Scholarship Collection Electronic Theses and Dissertations Fall 12-21-2018 Pyrrolo[2,3-d]pyrimidine Classical Antifolates for Targeted Cancer Chemotherapy- Applications of Bioisosteric and Regioisomeric Substitutions for Improved Tumor-Selectivity and Potency Manasa Punaha Ravindra Follow this and additional works at: https://dsc.duq.edu/etd Part of the Medicinal Chemistry and Pharmaceutics Commons Recommended Citation Punaha Ravindra, M. (2018). Pyrrolo[2,3-d]pyrimidine Classical Antifolates for Targeted Cancer Chemotherapy- Applications of Bioisosteric and Regioisomeric Substitutions for Improved Tumor-Selectivity and Potency (Doctoral dissertation, Duquesne University). Retrieved from https://dsc.duq.edu/etd/1747 This One-year Embargo is brought to you for free and open access by Duquesne Scholarship Collection. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Duquesne Scholarship Collection. PYRROLO[2,3-d]PYRIMIDINE CLASSICAL ANTIFOLATES FOR TARGETED CANCER CHEMOTHERAPY- APPLICATIONS OF BIOISOSTERIC AND REGIOISOMERIC SUBSTITUTIONS FOR IMPROVED TUMOR-SELECTIVITY AND POTENCY A Dissertation Submitted to the Graduate School of Pharmaceutical Sciences Duquesne University In partial fulfillment of the requirements for the degree of Doctor of Philosophy By Manasa Punaha Ravindra December 2018 Copyright by Manasa Punaha Ravindra 2018 PYRROLO[2,3-d]PYRIMIDINE CLASSICAL ANTIFOLATES FOR TARGETED CANCER CHEMOTHERAPY- APPLICATIONS OF BIOISOSTERIC AND REGIOISOMERIC SUBSTITUTIONS FOR IMPROVED TUMOR-SELECTIVITY AND POTENCY By Manasa Punaha Ravindra Approved August 28, 2018 _______________________________ ____________________________ Aleem Gangjee, Ph. D. Marc W. Harrold, Ph. D. Distinguished Professor of Medicinal Professor of Medicinal Chemistry Chemistry Graduate School of Pharmaceutical Sciences, Graduate School of Pharmaceutical Sciences, Duquesne University, Duquesne University (Committee member) (Committee Chair) _______________________ _______________________ Patrick T.
    [Show full text]
  • Evaluating Metabolic Stress Response Using Ultra Performance Liquid Chromatography-High Resolution Mass Spectrometry
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 5-2019 Evaluating Metabolic Stress Response Using Ultra Performance Liquid Chromatography-High Resolution Mass Spectrometry Eric David Tague University of Tennessee, [email protected] Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Recommended Citation Tague, Eric David, "Evaluating Metabolic Stress Response Using Ultra Performance Liquid Chromatography-High Resolution Mass Spectrometry. " PhD diss., University of Tennessee, 2019. https://trace.tennessee.edu/utk_graddiss/5348 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Eric David Tague entitled "Evaluating Metabolic Stress Response Using Ultra Performance Liquid Chromatography-High Resolution Mass Spectrometry." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the equirr ements for the degree of Doctor of Philosophy, with a major in Chemistry. Shawn R. Campagna, Major Professor We have read this dissertation and recommend its acceptance: Christopher Baker, Michael Best, Elizabeth Fozo Accepted for the Council: Dixie L. Thompson Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) Evaluating Metabolic Stress Response Using Ultra Performance Liquid Chromatography-High Resolution Mass Spectrometry A Dissertation Presented for the Doctor of Philosophy Degree The University of Tennessee, Knoxville Eric David Tague May 2019 Copyright © 2019 by Eric David Tague All rights reserved.
    [Show full text]
  • Midgut Metabolomic Profiling of Fall Armyworm (Spodoptera Frugiperda
    Insect Biochemistry and Molecular Biology 106 (2019) 1–9 Contents lists available at ScienceDirect Insect Biochemistry and Molecular Biology journal homepage: www.elsevier.com/locate/ibmb Midgut metabolomic profiling of fall armyworm (Spodoptera frugiperda) with field-evolved resistance to Cry1F corn T Heba Abdelgaffara, Eric D. Tagueb, Hector F. Castro Gonzalezb,c, Shawn R. Campagnab,c, ∗ Juan L. Jurat-Fuentesa, a Department of Entomology and Plant Pathology, University of Tennessee, Knoxville, 37996, USA b Department of Chemistry, University of Tennessee, Knoxville, 37996, USA c Biological and Small Molecule Mass Spectrometry Core, University of Tennessee, Knoxville, 37996, USA ARTICLE INFO ABSTRACT Keywords: Populations of the fall armyworm (Spodoptera frugiperda) have developed resistance to transgenic corn producing Spodoptera frugiperda the Cry1F insecticidal protein from the bacterium Bacillus thuringiensis (Bt). Resistance in S. frugiperda from Metabolomics Puerto Rico is genetically linked to a mutation in an ATP Binding Cassette subfamily C2 gene (SfABCC2) that Cry toxin results in a truncated, non-functional Cry1F toxin receptor protein. Since ABCC2 proteins are involved in active Resistance export of xenobiotics and other metabolites from the cell, we hypothesized that Cry1F-resistant fall armyworm Fall armyworm with a non-functional SfABCC2 protein would display altered gut metabolome composition when compared to susceptible insects. Mass spectrometry and multivariate statistical analyses identified 126 unique metabolites from larval guts, of which 7 were found to display statistically significant altered levels between midguts from susceptible and Cry1F-resistant S. frugiperda larvae when feeding on meridic diet. Among these 7 differentially present metabolites, 6 were found to significantly accumulate (1.3–3.5-fold) in midguts from Cry1F-resistant larvae, including nucleosides, asparagine, and carbohydrates such as trehalose 6-phosphate and sedoheptulose 1/7-phosphate.
    [Show full text]