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Proteomics & Bioinformatics conferenceseries.com 773rd Conference 7th International Conference on Proteomics & Bioinformatics October 24-26, 2016 Rome, Italy Posters Proteomics & Bioinformatics 2016 Page 91 Zubida Al-majdoub, J Proteomics Bioinform 2016, 9:11(Suppl) conferenceseries.com http://dx.doi.org/10.4172/0974-276X.C1.092 7th International Conference on Proteomics & Bioinformatics October 24-26, 2016 Rome, Italy Quantification of drug-metabolising enzymes in human liver microsomes: A comparison between label-free profiling and targeted quantitative analysis Zubida Al-majdoub University of Manchester, UK enonbiotic and drug-metabolizing enzymes (DMEs) are involved in the bioconversion of xenobiotics (including drugs, Xsynthetic chemicals and environmental pollutants) into inactive or active metabolites. In pharmacological therapy, bioconversion can either lead to detoxification or activation of the drug, which has implications on treatment effectiveness and toxicity. Quantitative profiling of the drug-metabolising sub-proteome can be used in the characterisation of liver drug metabolism profiles in individual patients which can be a major step towards stratified or personalized medicine. Immunoquantification and targeted proteomics approaches have traditionally been used to determine abundances of CYP and UGT enzymes; however, bias in the determination of absolute protein abundance between laboratories and methods has been demonstrated. This may be due to differences in methodological workflows or the choice of suitable and specific standards. Label-free analysis can provide a venue for a new methodological setup. Advantages of this type of approach include the possibility of quantifying a large number of proteins without the need for specific standards, allowing comprehensive description of dynamic changes of expression in the proteome under study. As an alternative approach to previously used methods, we aimed to apply a label-free proteomic strategy to quantify and assess the absolute expression of several CYP and UGT enzymes in microsomal fractions extracted from 27 human livers which have previously been characterised. Biography Zubida Al-majdoub has completed her Under-graduate studies in Tripoli University, Libya and worked as Teaching Assistant at the same university. She has completed her MPhil in Medicinal Chemistry from School of Pharmacy, University of Manchester, followed by PhD in Quantitative Proteomics under the supervision of Dr. Jill Barber and Professor Simon Gaskell. She is a Research Associate at School of Pharmacy. Her research work focuses on “Quantification of transporters in human brain”. [email protected] J Proteomics Bioinform 2016 Proteomics & Bioinformatics 2016 Volume 9, Issue 11(Suppl) ISSN: 0974-276X JPB, an open access journal October 24-26, 2016 Page 92 Valerie Morineaux et al., J Proteomics Bioinform 2016, 9:11(Suppl) conferenceseries.com http://dx.doi.org/10.4172/0974-276X.C1.092 7th International Conference on Proteomics & Bioinformatics October 24-26, 2016 Rome, Italy Mass spectrometry in forensic identification of two main potential agents of bioterrorism and warfare: Botulinum toxins and ricin Valerie Morineaux, J Enche and D Hilaire Direction générale de l’Armement, France rotein toxins such as botulinum neurotoxins and plant toxin ricin are considered as potential agents for bioterrorism and Pwarfare. Botulinum neurotoxins are produced by Clostridium botulinum commonly found in plants, soils, water and in the intestinal tract of animals. They act by blocking the release of acetylcholine, the principal neurotransmitter at neuromuscular junctions, causing muscle paralysis. Botulinum toxins are among the most poisonous substances. Ricin, on the other hand, is produced by the seeds of castor bean plant (Ricinus communis) which is used to process the castor oil. Ricin is very toxic against eukaryotic cells by inhibiting the protein synthesis and causing cell death. Unambiguous identification of these toxins is required not only for the implementation of effective countermeasures in case of terrorism event but also for law enforcement. This is the reason why we have developed a specific strategy for the detection and identification of botulinum toxins and ricin in complex matrices, by LC-QqQ-MS/MS method operating in Multiple Reaction Monitoring (MRM) mode. In order to be compatible with complicated samples, the mass spectrometry analysis was coupled with an immunocapture step. This method was successfully applied to the identification of botulinum toxins type A subtypes and ricin from complex matrices. Biography Valerie Morineaux is an Engineer in the Toxicology department, DGA, regarding NRBC (Nuclear, Radioactive, Biology and Chemical) risks. She has completed her PhD from DGA and Pasteur Institute. She has been working for several years on “The development of analytical methods in toxin identification”. [email protected] Notes: J Proteomics Bioinform 2016 Proteomics & Bioinformatics 2016 Volume 9, Issue 11(Suppl) ISSN: 0974-276X JPB, an open access journal October 24-26, 2016 Page 93 Zdenka Kristofikova et al., J Proteomics Bioinform 2016, 9:11(Suppl) conferenceseries.com http://dx.doi.org/10.4172/0974-276X.C1.092 7th International Conference on Proteomics & Bioinformatics October 24-26, 2016 Rome, Italy Nitric oxide synthases and acute sleep deprivation Zdenka Kristofikovaand Jana Sirova National Institute of Mental Health, Czech Republic myloid beta peptide and protein tau play a role in the development of Alzheimer disease. It is suggested that normal aging Ais the main risk factor and chronic sleep deprivation is the contributing risk factor here (e.g. via oxidative stress-evoked changes in amyloid beta and protein tau). A great attention is also focused on nitric oxide directly involved in sleep-wake cycle and Alzheimer disease pathogenesis. The purpose of the study is to evaluate changes in activities of nitric oxide synthases (neuronal, endothelial and inducible) in the right and left cortex of young or old rats exposed to increased locomotion (control experiments for the non- specific effects of the apparatus) or acute sleep deprivation (24 hours). In future, results will be compared with those obtained on young or old rats exposed to chronic sleep deprivation. We used Rat Forced Exercise Bed model 80805A*C apparatus (Campden Instruments Ltd.) based on the rotational movement of the activity wheel. Experiments were performed on young (3-4 months) and old (11-12 months) male Wistar rats. Activities of neuronal and endothelial synthases were significantly decreased in old compared to young control rats. The activity of inducible synthase was decreased in the left cortex of young rats exposed to increased locomotion. On the contrary, the activity of inducible synthase was increased in old rats exposed to acute sleep deprivation. The previous analysis of human autoptic brains revealed increased activities of all synthases especially in the left side of people with Alzheimer disease. It seems that acute sleep deprivation can evoke some changes in old rats similar to those seen in Alzheimer disease. Biography Zdenka Kristofikova is interested in problems of Alzheimer disease and normal aging for a long time. She focusses especially on “The analysis of the human or rat brain tissue and of cerebrospinal fluid”. Her main aims are “Evaluations of new animal models of Alzheimer disease or of new biomarkers of Alzheimer disease sensitive to changes in early stages of the disease”. [email protected] Notes: J Proteomics Bioinform 2016 Proteomics & Bioinformatics 2016 Volume 9, Issue 11(Suppl) ISSN: 0974-276X JPB, an open access journal October 24-26, 2016 Page 94 Ramona Bosch, J Proteomics Bioinform 2016, 9:11(Suppl) conferenceseries.com http://dx.doi.org/10.4172/0974-276X.C1.092 7th International Conference on Proteomics & Bioinformatics October 24-26, 2016 Rome, Italy Expression and purification of membrane scaffold proteins for the design of discoidal phospholipid bilayer nanoparticles Ramona Bosch University of Hohenheim, Germany he expression and purification of recombinant proteins is an everyday business. Years ago, however, protein production was Tdifficult, time consuming and remained mostly in the domain of experts. The progress of simple, commercially available systems made the technology more widespread and led to an increase in protein production. In this work, a derivate of the human apolipoprotein A-1, the Membrane Scaffold Protein (MSP) is produced biotechnologically in a lab scale bioreactor. The MSP is a genetically engineered protein which has the notable feature to self-assemble into discoidal nanoparticles in the presence of synthetic phospholipids. These so-called nano discs have become increasingly important in the last few years e.g., for the study of membrane-associated proteins. In previous studies, nano discs are assembled by adding a micelles-organized detergent-phospholipid mixture to an aqueous system containing the MSPs. Upon removal of detergent, 10 nm diameter particles are formed. Unfortunately, the resulting nano discs are disordered in this solution and therefore further processing into an ordered and directed membrane cannot be easily achieved. The aim of this work is to create biomimetic membranes consisting of cross-linked nano discs e.g., the translocon SecYEG as part of an embedded protein complex for an active biological transport of potential target proteins. Now, the approach should be replaced by nano patterning
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