6^th International Conference on Biomarkers & Clinical Research September 01-02, 2015 Toronto, Canada

Jing Zhang is a recent PhD graduate from Carleton University under Dr. Kenneth Storey’s supervision and joined Dr. Valerie Langlois’ group as a Post-doctoral fellow in the fall of 2013. His field is molecular physiology and previous work includes investigating molecular mechanisms behind survival adaptions under extreme environmental conditions in various stress tolerant animal models. Currently, he is focusing on exploring the potential of hair follicle as a diagnostic tool for military activity-related medical conditions including traumatic brain injury (TBI) and operational stress disorders using transcriptomic approaches.

Military personnel commonly exposed to operational stressors (example, intense physical exertion, sleep deprivation and fatigue) during both training activities and combat operations, which are often, associated with mental health disorders including depression and post-traumatic stress disorder. As psycho-physiological changes driven by molecular mechanisms, it is useful to assess corresponding adaptations in gene expression profiles for a better understanding of such responses. Thus, we explored human hair follicles as a robust and accessible biomarker system for both characterization and diagnostic purposes. Four healthy male volunteers performed a 2-week period of high-intensity interval training (HIIT) as an easily quantifiable and reproducible physical stress model. RNA-sequencing was conducted on an Illumina MiSeq platform using the total RNA libraries prepared from the hair follicles collected from the vertex area of the scalp pre- and post-HIIT. The resulting RNA sequences aligned to the human genome. Differential expression (DE) analyzed for protein coding RNA (mRNA), miRNA and long non-coding RNA (lncRNA). Gene ontology (GO) term analysis also conducted. Based on the DE mRNA results, the GO term enrichment showed 192 positively affected terms (example, hormonal control), while 387 (example, immune system process) were negatively enriched upon HIIT. Regarding to miRNA, 48 related GO terms (example, antioxidant response) enriched. We also identified 249 differentially expressed lncRNAs including those involved in RNA processing (example, RMRP). Overall, this study demonstrates that human hair follicles are a valuable resource inproviding molecular signatures linked to operational stress disorders.

The incidence of allergy/immunotoxicity-related post-market adverse events associated with medical devices is increasing. Biomarkers are commonly used in toxicology for risk assessment and clinically as diagnostic and monitoring tests. We developed a new in vitro model where human peripheral blood mononuclear cells (PBMC) isolated from four healthy donors were used to identify biomarkers that specifically respond to metallic allergens. Cells were exposed to well-known human metallic allergens and non-allergens for 24 hours. The cell surface markers were measured by employing flow cytometry. A primary goal was to determine whether the two cell types are transferable from the dendritic cell (DC) to the PBMC due to the clinical relevance of the PBMC. Of the 12 cell surface markers following the first tier selection, the expression of CD1X and CD86 on PBMC was confirmed and 3 other novel biomarkers were discovered (CD2X, CD3X and CD4X). The expression of CD1X on PBMCs was down-regulated significantly following exposure to three metallic allergens (cobalt (II) chloride, nickel (II) sulfate, potassium dichromate (VI)) while the expression remained the same when exposed to two metallic non-allergens (lead (IV) acetate, magnesium (II) chloride) compared to untreated PBMCs. Data from the all four donors showed the same pattern and the statistical value of CD1X alone yielded a p value<0.0001 and an accuracy greater than 95% based on receiver operating characteristic (ROC) analysis. We found the consistency of CD1X performance in PBMC even greater compared to that in DC. Data indicate that CD1X shows promise for use as a pre-clinical biomarker for screening potential allergenic responses to metal-containing devices. Further validation studies are planned.

Kalpana Kalyanasundaram Bhanumathy has completed her Master’s and PhD degree in Biotechnology from Annamalai University, India. During her Master’s, she has received several awards for her academic excellence and has bagged the “Best women student award” among the Departments in the Faculty of Science. Her PhD research was related to field of “Radiation Oncology” and she was the recipient of the “Senior Research Fellow (SRF) Award” instituted by the “Indian Council of Medical Research (ICMR)”, Government of India during her PhD programme. After completing her PhD, she moved to Saskatoon, Canada to pursue her Post- doctoral work in the field of Immunology under Dr. Jim Xiang, Senior Scientist and Professor at the University of Saskatchewan. She was then awarded the prestigious “Terry fox Post-doctoral fellow” - by the Saskatchewan Health research foundation (SHRF), Saskatchewan, Canada. She also received various travel awards to present her work in International conferences. She has published eight papers as first author and two papers as co-author in peer reviewed international journals.

Adoptive transfer of CD8+ T-cells specific for tumor-antigens is an attractive strategy for anti- tumor therapy. In this study, the subsets TA and TB were used to represent the population of CD8+ T cells generated by culturing the respective cells with irradiated dendritic cells (DCs) pulsed with ovalbumin (OVA) protein and transfected with adenoviral vector constructs. Naïve OVA specific CD8+ T cells were isolated from the spleen of OVA-specific T-cell receptor transgenic OTI mice. The subsets TA and TB were then generated by activating the population of CD8+ T cells with OVA-pulsed DCs transfected with IL-6-expressing adenoviral vector (AdVIL- 6) or the control vector (AdVNull). To assess their in vivo immunotherapeutic effects, TA- or TB- cells were i.v. transferred into C57BL/6 mice bearing EG7 thymoma. TA-cells displayed higher level expression of CD62L, IL-7R, FasL, perforin and CCR6; exhibited more potent in vitro cytotoxicity to OVA-expressing EG7 thymoma cells via perforin- and Fas/FasL-mediated apoptosis than TB-cells. CD8+ T-cell survival was kinetically analyzed in C57BL/6 mice adoptively transferred with TA- or TB-cells by flow cytometry. We found that the adoptively transferred TA-cells had prolonged survival and enhanced T-cell memory development, compared to TB-cells. In addition, TA-, but not TB-cells were able to eradicate well-established EG7 thymomas in all 8/8 tumor-bearing mice. Our data suggested that AdVIL-6-transfected DC- stimulated CD8+ T cells with potent cytotoxicity and survival advantage may serve as an effective adoptive CD8+ T-cell immunotherapy strategy for anti-tumor treatment.

Ting-Yu Chang has obtained his PhD degree in July 2011 from Institute of Microbiology and Immunology from National Yang-Ming University. He is now a Post-doctoral Research Fellow in Dr. Hsei-Wei Wang’s lab and is in charge of dry-lab NGS data analysis and translational research. He is an experienced Molecular Biologist and Bioinformatitician with a great interest in genomics as well as a Statistical Genetics Analyst with extensive bioinformatics skills. He involves in a wide range of research projects from a transcriptomic viewpoint-viral-host interaction, cancer pathology, cancer stem cell biology, endothelial cell biology and more.

MicroRNAs (miRNAs) have emerged as master regulators of angiogenesis and other cancer-related events. Discovering new microRNAs regulating angiogenesis (angiomiRs) will eventually help in developing new therapeutic strategies for tumor angiogenesis and cardiovascular diseases. Kaposi’s sarcoma (KS) which is induced by the etiological infectious agent KS-associated herpes virus (KSHV) is a peculiar neoplasm expressing both blood and lymphatic endothelial markers and possessing extensive neovasculature. Using KSHV and its viral proteins as baits will be an efficient way to discover new angiomiRs in endothelial cells. Kaposin B is one of the few latent viral genes and is expressed in all KSHV tumor cells. Since kaposin B is a nuclear protein with no DNA-binding domain, it may regulate gene expression via incorporating into a transcription complex. We found here that cMyc and kaposin B form a transcription complex to bind to microRNA promoters. By small RNA sequencing (smRNA-Seq) we disclosed that cMyc and kaposin B co-repress anti-angiogenic miRNAs such as miR-221/-222 while inducing pro-angiogenic miRNAs such as miR-210. 64.9% (213/328) of kaposin B up-regulated miRNAs and 62.0% (181/292) of kaposin B down-regulated miRNAs were regulated by cMyc. Our study thereby illustrates new angiomiRs regulated by cMyc and a cMyc-orientated program is coordinated by kaposin B in KSHV-infected cells.

Rohan R Ravindranath completed his BSc (Hons) from the University of Toronto in Chemistry and Biotechnology. Currently, he is pursuing his MSc in Chemistry under the joint guidance of Professor Michael Thompson and Professor Alexander Romaschin at the University of Toronto and at St. Michael’s hospital. He has presented his past research at various local conferences and has been awarded a best poster prize.

Nanomedicine continues to be an emerging discipline, providing novel solutions to diseases that otherwise could not be treated effectively. The development and use of self-assembled monolayers and adlayers for biomedical applications has allowed bio-incompatible materials such as stainless steel and cobalt-chromium, to be used as effective medical devices. In terms of stainless steel, this material has been widely used to develop stent implants. Stents are expandable tubes used to open up a restricted artery. Conventional implantation of a bare metal stent can cause renarrowing of the artery over time restenosis, due to an immune response launched by the body towards the “foreign” stent. As a result, improving the biocompatibility of stainless steel is thought to be critical for creating a stent that is not rejected by the body. Benzothiosulfonate (BTS) and Pentafluorophenyl Ester (PFP) are two chemical molecules that have previously been used as surface modifiers of stainless steel and quartz. BTS was recently used as a surface coating on stainless steel to bind antibodies, which could then in principle be used to bind specific biomarkers on circulating endothelial cells. For this stent application, it is important that the surface coatings show minimal chemical change over time, when placed in a physiological environment. This investigation involved assessing the long-term stability of BTS and PFP coatings on stainless steel. Surface characterization techniques like X-ray photoelectron spectroscopy, contact angle goniometry and atomic force microscopy were used to analyze the integrity and composition of the adlayers upon immersion in physiological buffer solution.

Dmitrii Cherepakhin is a student of I M Sechenov First Moscow State Medical University. She is Member of Young professional group in structure of EPMA. She was a participant and speaker of two international congresses about Predictive-Preventive and Personalized Medicine (PPPM) in Bonn and in Brussels. She is an Author of serial articles about PPPM in cardio-vascular pathology and nowadays she works in genetic and in predictive and preventive medicine.

Cardiovascular Diseases (CVD) are the most common cause of death worldwide. Today CVDs accounts for 30% of deaths worldwide. Cardiovascular disease has long been at the forefront of gene testing in the clinic, and this trend is likely to continue. New disease genes add to our mechanistic understanding of the biology of the disorders. Building a picture of the comprehensive genetic architecture of each syndrome will eventually aid in diagnosis or prognostication, as well as drive the basic science that generates novel therapies. While clinical gene identification is currently of utility mainly in the restriction of cascade screening efforts, this will change as pathway focused therapies become available. Genetic testing in hypercholesterolemia, which previously had been of no obvious clinical importance given the precision of risk prediction with plasma cholesterol measurements and the empiric nature of subsequent management, is likely to become much more prevalent if PCSK9 inhibitors demonstrate efficacy. Similarly, imminent clinical trials in subsets of hypertrophic cardiomyopathy e.g. genotype positive–phenotype negative individuals or those with RAS-MAP kinase pathway mutations, dilated cardiomyopathy e.g. lamin a/c mutations or even allele-specific silencing may soon mandate genetic testing in these disorders. The rapid growth of genetic testing in cardiovascular disease is an important paradigm for the broader application of genetic testing across medicine, where the initial presentation may often be an avoidable sudden death.

Samy Otero González is close to complete his studies on Informatics Engineering next year at the Polytechnic University José Antonio Echeverría (CUJAE). He developed a passion for Bioinformatics when working as a Chemical technician in the Center for Genetic Engineering and Biotechnology (CIGB, 2002-2014). So he decided to work on his thesis at CIGB’s Systems Biology department in close collaboration with the logistic department at CIMAB.

PD2 process is a web-based tool implemented in PHP, JavaScript and Perl for facilitating the functional interpretation of proteomics data. Through a modular approach this tool assists the processing of comparative proteomics, pull-down and phosphoproteomics experiments results. It provides and interactive environment where the researcher can visualize and/or compare sets of quantitative data as well as protein functional information for post-translational modifications, subcellular localization, gene ontologies, etc. The tool also provides links to several biological databases to somehow centralize the information available in these resources. In conclusion, PD2Process constitutes an alternative for storing, visualizing and analyzing proteomics data that focus on the functional description of the identified/quantified proteins and in the case of phosphoproteomics of the identified phosphosites.

Teresa Núñez de Villavicencio Díaz has completed her licentiate studies in Biochemistry at the Faculty of Biology, University of Havana, (with distinction) and obtained a Master’s degree in Biotechnology, Biopharmaceutical Research, at the Center for Genetic Engineering and Biotechnology. She is a researcher with three years of experience in systems biology, proteomics, bioinformatics and molecular oncology. She published a paper at current topics in medicinal chemistry which already has a citation and more than a thousand downloads at ResearchGate.

The functional interpretation of data is an inherent task associated to genomics and proteomics. The quality of the analysis is dependent on the workflow designed by the researcher and the bioinformatics tools chosen to integrate it. Currently it is imperative to understand that no analytical strategy or bioinformatics tool by itself is capable of extracting all the information covered, for example, in a proteomics experiment. Hereby we seek to provide the researcher community with four groups of different but complementary bioinformatics tools that can be used when performing this type of analysis. In addition, we describe an integrative bioinformatics workflow in the context of biomarkers discovery and the study of drugs’ action mechanisms.

Background: According to the world health organization (WHO) statistics, there is about one third of male adult smokers, which can lead to four million deaths per year. Tobacco addiction could be potentiated by monoamine oxidase (MAO) inhibition as several MAO inhibitors were identified in tobacco smoke. Bupropion exhibits antismoking activity in addition to its known anti-depressant effect. The primary goal of this project was to synthesize new bupropion analogs with potent MAO inhibitory activity.
Methods: We synthesized new bupropion analogs using 3`-chloropropiophenone as a starting material. The first step was the synthesis of -bromoderivative using bromine in glacial acetic acid followed by amination with different amines. We converted these obtained products immediately to the corresponding fumarate salts. We monitored all reactions by thin layer chromatography (TLC) and purified them by crystallization, solvent-solvent extraction and column chromatography. We elucidated the structures of the prepared compounds by nuclear magnetic resonance (NMR), Fourier-transform infrared spectrophotometry (FT-IR) and mass spectrometry (MS). Nowadays, the MAO-B inhibitory activity is being evaluated against recombinant human MAO-B using 96-well microtitre plate reader.
Results: We prepared several bupropion analogs and confirmed the structure of the purified compounds using with FT-IR and LC/MS. Currently, we are investigating the prepared compounds in relation to their MAO inhibition activity.
Conclusion: We successfully were able to achieve the synthesis and structural elucidation of the several compounds. These derivatives, which have potent MAO inhibitory activity, will potentiate the anti-smoking effect of the Bupropion and will have a pronounced impact on the future of the smoking cessation pharmacotherapies alternatives.

Sareh Keshavarzi completed her PhD at Shiraz University of Medical Sciences. She also worked as an Assistant Professor. Her research efforts to date have led her to publication of 32 refereed papers, 6 technical papers, 3 major reports or published abstracts and 19 presentations of her research at professional meetings at the regional, national and international levels.

In the experiments on Quantitative Trait Loci (QTL) mapping, typically phenotypic data on multiple correlated traits are collected. However, most genetic association analyses consider traits separately and ignore potential correlations among the traits. A Seemingly Unrelated Regression (SUR) system that was introduced by Zellner in 1961, includes several individual relationships that are linked by the fact that their disturbances are correlated. In this study, we developed SUR model as a more efficient multivariate model for correlated traits to evaluate the extent of common genetic influence on five plasma lipid fractions; i.e., Low Density Lipoprotein Cholesterol (LDL-C), High Density Lipoprotein Cholesterol (HDL-C), triglycerides, Apolipoprotein A1 (ApoA1), and Apolipoprotein B (ApoB). By jointly analyzing multiple correlated continuous traits, this model leads to more efficient estimators by combining information on different equations compared to the traditional multivariate model. Moreover, in contrast to the traditional multivariate model, different traits can have different sets of predictors through SUR model. Simulated data from genetic analysis illustrate application of the proposed method. The analysis was performed with knowledge of the simulation model. The proposed model can examine multiple single-nucleotide variations simultaneously and incorporate family-specific, subject-specific, or time-varying covariates. Analysis of the simulated data illustrated the applicability of the proposed approach in modeling genetic loci associated with multivariate outcomes of plasma concentrations of LDL-C, HDL-C, triglycerides, ApoA1, and ApoB among women in genome-wide analysis. Results from simulation studies also showed that the estimators from the proposed model were more efficient compared to those based on ordinary least square. Furthermore, comparative performances of the estimators were investigated on the basis of the standard errors of the parameter estimates and the results indicated that the estimators performed better as the sample size increased.

I am studying master of genetics in azad university.I am currently employed by Sarem hospital in Tehran and work in genetics lab,in PGD (preimplantation genetic diagnosis) section and also I am interested in biomarker blood cell research.

Background: Breast cancer is leading cause of cancer death in women worldwide but early detection is correlated free survival. Epigenetics particularly DNA methylation has recently been shown to be important in breast cancer initiation. We examined the association between promoter hypermethylation level of SFRP1 and CDH1 genes which are antagonist in WNT signaling and cell-cell adhesion respectively in peripheral blood leukocyte DNA and breast cancer among Iranian women.
Methods: We conducted a hospital-based case-control study of 30 patients aged less than 40 years with newly diagnosed sporadic breast cancer, no history of any therapy and 30 matched controls from medical checkup examinees in Imam Khomayni Hospital. Promoter methylation levels in leukocyte DNA were measured by MeDIP-Realtime Assay.
Results: Out of 30 breast cancer patients, 16 (53.3%) manifested various levels of SFRP1 and 1 (3.3%) of CDH1 methylation. However, no methylation was found in 30 controls.
Interpretation: These findings suggest that the SFRP1 promoter methylation level of peripheral blood leukocyte DNA is low in patients with breast cancer and may be a potential biomarker for breast cancer risk but highly methylated CDH1 promoter may be related to metastatic tumors.

The aim of this work is to develop and validate a dissolution method for 150 mg Tolperisone hydrochloride film coated tablets using a UV spectroscopic method. Tolperisone HCl, frequently used as centrally muscle relaxants, has no dissolution method in its monograph in Japanese pharmacopeia. After test sink conditions, dissolution medium, the best conditions were: Basket apparatus at 75 rotations per minute (rpm) stirring speed, 0.1 N HCl as dissolution medium volume of 900 ml. The quantitation method was also adapted and validated. More than 95% dissolution was achieved over 30 min in the basic one. The dissolution profile of three marketed brands of Tolperisone HCl 150mg tablets have been evaluated using dissolution test in 0.1N HCl media with the aim to assess bioequivalence. The dissolution test developed was adequate for its purpose and could be applied for quality control of Tolperisone hydrochloride tablets. The in vitro release profiles of various tests were compared for their similarity using model dependent and model independent methods. Results of this test indicate that in most cases dissolution profiles of the different marketed brands were significantly different from each other. The objective of this study was to assess the interchange ability of the available Tolperisone HCl products on the basis oftheir in vitro dissolution characteristics using USP Apparatus2 (Basket type).

Jinesh Kumar Patel is an Assistant Professor in Department of Biochemistry, M.G. Science Institute, Gujarat, India with five years of teaching and research experience. He has completed his Bachelor’s, Master’s from the Gujarat University and PhD from HNGU. He has made his significant contribution in the field of biochemistry and has published 5 papers in reputed journal.

Till date no analytical method has been developed for estimation of Amlodipine Besylate (AML) and Indapamide (IND) simultaneously in combined dosage form. Thus, the objective of present work is to Develop and validate simple, precise, accurate UV-Spectrophotometric methods and Spectrofluorometric method for simultaneous estimation of AML and IND in their combined dosage form. For Absorption correction method Linearity of AML and IND was found in 14-38 μg/ml and 4-12 μg/ml respectively, while for Dual wavelength method Linearity of AML and IND was found 14-44 μg/ml and 4-12 μg/ml respectively. In Spectrofluorometric method Linearity of AML and IND was found 0.2-3.4 μg/ml and 0.5-6.5 μg/ml, respectively. Precision for all mentioned methods was found to be less than 2% and Accuracy was found to between 98-102%. All methods were found to be simple, precise and accurate. All methods can be successfully used for routine simultaneous analysis of AML and IND in pure and combined dosage forms.

Zahra Jalali Sefid Dashti has completed her PhD at the South African National Bioinformatics institute (SANBI), University of the Western Cape. She is currently a Post-doctoral researcher at the same institute. Her research expertise is in the next generation sequencing data analysis of infectious diseases such as sleeping sickness and tuberculosis.

Tuberculosis (TB) is an infectious disease caused by bacteria belonging to the Mycobacterium tuberculosis complex (MTC). The global burden of disease based on the World health organization (WHO) estimates suggests 9.4 million new TB cases worldwide, with the majority of the estimated cases occurring in Africa and Asia. In spite of the existing challenge with the multidrug resistant (MDR) TB, TB control faces another dilemma; extensively drug resistant (XDR) TB a situation where the pathogen is resistant to the four most effective anti-TB drugs rifapincin, isoniazid, an aminioglycoside and a fluoroquinolone. Deciphering the molecular basis of drug resistance has become a research priority, as this information will guide the development of new molecular based diagnostics as well as providing insight into new drug targets. In line with this, we carried out the assembly and functional annotation of a relatively large dataset with 400 M. tuberculosis genomes using reference based assembly approach. Variant calling methods including GATK (Haplotypecaller), Samtools and FreeBayes were used to identify SNPs that map to specific phenotypic categories such as XDR, MDR, and drug susceptible. This data provides a knowledge base for drug resistance marker discovery.

Lamia Beghenna has completed his State Engineer in Chemistry from Badji Mokhtar University, Annaba (Algeria) and attended Master -2- (Euro-Maghreb) in Biotechnology, quality, security and recovery in photoproducts of health and nutrition of environment from Badji Mokhtar University, Annaba (Algeria).

Brassica oleracea, var. italica is generally known as an alimentary plant but it has some medicinal uses. The plant collected from of South Algeria, is phytochemical screened. Most of interest of corrent research relates to study the natural antioxydant molecules. Our work aim is to study a phytochemical and antioxidant activity of Brassica oleracea, var. italica leaves, and stems. The phytochemical tests realized can be to highlight the flavonoïdes, tannins, sterols and phenols in the two parts of the plant. The antioxidant activity of methanol-water (70:30) extract of leaves and stems were evaluated using DPPH test (2, 2-diphény 1- picryhydrazyl) expressed by Vitamin C Equivalent Antioxidant Capacity (VCEAC). In addition, their total phenolic compounds (TPC) and total flavonoid compounds (TFC) were measured. The VCEAC values for the two parts of plant are 0.633 g/100g for stems and, 89.6 g/100g for leaves of vitamin C equivalents (VCEAC)/100 g of dry weight.