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Day 1 : Apr-15-2014
Keynote Forum
Biography:
Sergey Suchkov graduated from Astrakhan State Medical University and was awarded with MD.In 1985, Suchkov obtained his Ph.D. As a Ph.D. student of the I.M. Sechenov Moscow Medical Academy and Institute of Medical Enzymology, USSR Academy of Medical Sciences, Moscow, Russia. In 2001, Suchkov finished the PostDoc Research Fellowship Program and maintained his Doctor Degree at the National Institute of Immunology, Russia. From 1987 through 1989, Dr. Suchkov was a senior Researcher, Lab of Developmental Immunology, Koltzov Institute of Developmental Biology, USSR Academy of Sciences to deal to developmental immunology. From 1989 through 1995, Dr. Suchkov was being a Head of the Lab of Clinical Immunology and Im-munobiotechnology, Helmholtz Eye Research Institute in Moscow. From 1995 through 2004, Dr. Suchkov was being a Chairman of the Department for Clinical Immunology, Moscow Clinical Research Institute (MONIKI) and the Immunologist-in-Chief of the Moscow Regional Ministry of Health. At present, Dr Sergey Suchkov, M, Ph.D., is Professor in Immunology, Department of Pathology, School for Pharmacy, I.M. Sechenov First Moscow State Medical University, Dean of the Department (Faculty) of The PPPM Development, and the First Vice-President of the University of World Business, Politics and Law and Secretary General, United Cultural Convention (UCC), Cambridge, UK
Abstract:
A new systems approach to disease to pay its crucial attention on the trend would result in a new branch in the healthcare services, namely, predictive, preventive and personalized medicine (PPPM).Meanwhile, all chronic disorders develop gradually over a period of time to take years for a process to reach a level where it could be diagnosed definitively and treatment initiated properly and in time before changes are irreversible! And to achieve the implementation of PPPM concept, it is necessary to create a fundamentally new strategy based upon the subclinical recognition of biomarkers and biopredictors of hidden abnormalities long before the disease clinically manifests itself. PPPM is thus a medical model being tailored to the individual and dictates a construction of PPPM algorithms to diagnose, to predict, and to prevent in time whilst following a concept of biomarkers impact into the daily practice!The key benefits of PPPM include new abilities: (i) to detect disease at a subclinical stage, when it is easier and less expensive to treat effectively; (ii) to stratify patients into groups that enable the selection of optimal preventive treatment; (iii) to reduce adverse drug effects by more effective early assessment of individual drug responses; (iv) to improve the selection of new molecular targets for drug discovery; (v) to shift the emphasis from illness to wellness.The first discriminatory step illustrating the PPPM-oriented survey is estimating of the correlation strength between genetic polymorphism and risks of the disease, and subsequent construction of the groups at risks. Those goals can be solved by using of BioChip methodology (each disease has specific biomarkers and thus the individual fingerprints). As a result, a patient becomes a data carrier, i.e., he/she knows about possible risks of a disease, and the physician can reasonably select of preventive protocol, proceeding from the assays made. Individuals, selected at the first stage, undergo the second phase of the survey, which uses a panel of phenotypic biomarkers and biopredictorsIt would be extremely useful to integrate data harvesting from different databanks for applications such as prediction and personalization of further treatment. So PPPM whilst utilizing a highly promising concept of biomarkers and biopredictors, would offer great and real challenge for the future, and next generations will speak about the XXI century as a time, when healthcare services became predictive and preventive, and its outcomes - secured and guaranteed!






Keynote Forum

Pavel Vodicka

Institute of Experimental Medicine, Czech Republic

Keynote: Biomarkers in ethiology, onset and prognosis of gastrointestinal cancers
Biography:
Pavel Vodicka graduated at the Medical Faculty, Charles University, Prague and in 1986 obtained Ph.D. in biochemistry. He worked as postdoctoral fellow at the Finnish Inst. Occupat. Health, Helsinki, Finland (1987-1990) and as visiting scientist at Karolinska Institute, Huddinge, Sweden (1990-1993). Since 2002 he heads the Dept. Molec. Biol. Cancer, Inst. Exper. Medicine, Acad. Sci., Prague, Czech Republic. Pavel Vodicka has published more than 130 (total IF 552.4, 2650 citations a HI 31) articles. Since 2004, his main research topics are focused on the DNA and chromosomal damage and DNA repair functional tests in humans and on transient biomarkers in the onset of gastrointestinal cancers. In 2012 he edited the Special Issue in Mutagenesis (http://mutage.oxfordjournals.org/content/27/2.toc), entitled Colorectal Cancer-Current Insights into Susceptibility.
Abstract:
According to the NCI definition biomarkers represent biological molecules in blood, other body fluids or tissues that are signs of normal or abnormal process or disease (e.g. cancer). They shall objectively differentiate and evaluate normal biological processes from the pathological ones. Additionally, biomarkers should map pharmacological response to therapeutic intervention. In defining the disease biomarkers may be arbitrarily classified into following, rather clinical, categories: i)biomarkers for diagnosis/screening, ii) biomarkers for evaluation of progression, iii) biomarkers of prediction of response and iv) overall prognosis of the disease. We can also stratify biomarkers into more theoretical groups, comprising biomarkers of genomic landscape of cancer, biomarkers defining functions of substantial biological systems (or whole organism), biomarkers of epigenetic regulations, biomarkers of cancer phenotype, biomarkers of treatment response and biomarkers of immune response. In case of colorectal cancer biomarkers describing the microenvironment are essential. The whole process of biomarke\'s development utilization has recently been described by (Taenzer et al. 2013), however, by using biomarkers following preconditions should be born in mind: their sensitivity, validity, reproducibility, availability, informativness, cost effectivness and complexity versus interpretability. Cancer undoubtedly exhibits a complex, multifactorial origin, comprising a plethora of genetic and environmental/life style factors acting in interaction. Th e scope of selected biomarkers should reflect this complexity. Implementation of proof of principle and logical chain in employed biomarkers is of utmost importance. Gastrointestinal (GIT) cancers, colorectal and pancreatic cancers in particular, represent serious health problems worldwide, irrespective of the stratification for the developmental status of the particular country. Incidence of GIT malignancies takes four to five places in the top ten cancer locations and the same tendency is reflected in the mortality. Due to these sombre statistics we directed our effort to the investigations of these malignancies and several examples on the use of disease biomarkers will be provided. One of the most prominent aims is a dissection of validated, early and reliable transient biomarker(s) predicting in advance the onset of cancer.






Keynote Forum

Ondrej Topolcan

Charles University in Prague, Czech Republic

Keynote: Serum levels of markers in early detection of prostate (pilot study)
Biography:
Ondrej Topolcan is the head of Immunoanalytical Laboratory Medical School Pilsen, Department of Nuclear Medicine Faculty Hospital Pilsen, Charles University, Prague. He is also Deputy Director of Faculty Hospital Pilsen for research and science. He is national representative and member of National Board European Association for Predictive, Preventive & Personalized Medicine (EPMA) and member of Academic Board EPMA. His research is particularly dedicated to the biomarkers related to endocrinology, oncology and internal diseases. He has been organizing National Immunaoanalytical Conference in the Czech Republic as well as many international meetings related to the biomarkers. He was awarded Carl R. Jolliff Award by AACC in 2011.
Abstract:
Background: Monitoring changes in the levels of biomarkers PSA, %freePSA, [-2]proPSA and calculation of PHI in the diagnostic algorithm of early prostate cancer. Patients and Methods: The Immunoanalytical Laboratory of University Hospital in Pilsen examined sera of 76 patients from the Urology Department of the University Hospital with suspected prostate cancer who have undergone TRUS biopsy. We assessed the levels of PSA and, when the interval of PSA was between 0-30 ng/mL, we also assessed the levels of freePSA, [-2]proPSA and we calculated %freePSA and Prostate Health Index (PHI). The monitored biomarkers were measured using the chemiluminescent DxI 800 instrument (Beckman Coulter, USA). All statistical analyses were calculated using the SAS version 9.2software. Results and discussion: We found statistically significant increased levels of [-2]proPSA and PHI in patients diagnosed with prostate cancer by prostate biopsy vs. patients with benign prostate hypertrophy ([-2]proPSA median 14 vs. 27 pg/mL, PHI median 35 vs. 77). On the contrary, we did not find any significant difference in tPSA and %freePSA (median tPSA 7.1 vs. 7.7 ng/mL and %freePSA 16 vs.11.4%). Analyte Units Reference range Instrument tPSA ng/mL Age adjusted DxI 800, Beckman Coulter, USA fPSA ng/mL - DxI 800,Beckman Coulter, USA n[-2] ProPSA pg/mL - DxI 800,Beckman Coulter, USA %fPSAm % 20-100% calculated PHI - >40 calculated Analyte (units) PSA (ng/mL) proPSA (pg/mL) %freePSA n(%) PHI (-) Diagnosis Count (N) Median (Min – Max) Median (Min – Max) Median (Min – Max) Median (Min – Max) Benign histology 50 7.13 (2.58 – 30.3) 13.7 (3 – 110) 16 (1.8 – 48) 35 (13 – 155) Malignant histology 26 7.68 (4.64 – 30.5) 22.5 (8.2 – 111) 11 (4.3 – 31.7) 69 (42– 117) Conclusion: The assessment of [-2]proPSA and the calculation of PHI appear to be of great benefit for a more accurate differential diagnosis of benign hyperplasia.






Track 1-1 Protein Biomarkers
Track 1-4 Cancer Biomarkers
Session Chair

Claude Prigent

University of Rennes, France




Session Co-Chair

Wancai Yang

University of Illinois at Chicago, USA




Session Introduction
Biography:
He is the Director of Research CNRS, Director of the Institute of Genetics and Development of Rennes, UMR 6290 CNRS/Univ Rennes1, France and also the Director MRic: Microscopy–Rennes Imaging Center, Head of the Cell Cycle team. He is currently studying the mechanisms that regulate different aspects of mitosis. He is particularly interested in the main mitotic structures, which are the centrosomes, the mitotic spindle and the chromosomes.
Abstract:
Aurora-A kinase is involved in many events controlling cell division specifically during mitotic progression. The protein is overexpressed in various cancers, and its gene amplified in some of them. As many protein kinases, Aurora-A must be phosphorylated in its activation loop on T288 to be active, and an antibody detecting phospho-T288 is now available and often used to estimate Aurora-A activity in cells. He will demonstrate that this phosphorylation event cannot be used as readout to measure Aurora-A activity in vivo. His laboratory has set up a new approach to specifically inhibit Aurora-A activity in cells. He will demonstrate how this approach can be used to estimate the specificity of Aurora-A inhibitors developed by pharmaceutical companies to be used in chemotherapy.






Biography:
Our laboratory aims to develop a better understanding of the processes of liver carcinogenesis in human hepatocellular carcinoma, focuses on expression profiles-based molecular genetic studies. Specifically, 1. We perform functional genomic studies to decipher the molecular differences between cancerous and normal liver tissues to identify novel diagnostic and prognostic biomarkers relating to human hepatocellular carcinoma. 2. To better understand the molecular roles of these biomarkers in the carcinogenesis of human hepatocellular carcinoma. 3. To design and build novel diagnostic platforms for human hepatocellular carcinoma using these novel biomarkers. 4. To design and perform animal experimentation to validate novel viral- and drug-mediated therapies for human hepatocellular carcinoma. 5. To design and generate cancer-specific vaccines for human hepatocellular carcinoma.
Abstract:
Tumor recurrence and metastases are the major obstacles to improve the prognosis of patients with hepatocellular carcinoma (HCC). In this presentation, we will describe the identification of blood-based diagnostic and prognostic biomarkers that surpass existing clinicopathologic factors in gauging the risk of early recurrent disease in patients with HCC and the development of HCC in high-risk CHB patients. To identify novel molecular signatures associated with HCC recurrence and metastases, we have established an expression database of HCC patients using Affymetrix gene chips. Samples from patients with early recurrent disease were compared with non-recurrent human HCC tissue samples (recurrence is defined as recurrent disease occurring within a 2-year time point of the original treatment). Novel biomarkers associated with early HCC recurrence have been characterized and are being employed as diagnostic markers for early recurrent disease. Metastasis that is responsible for the majority of cancer-related deaths is initiated by cancer cells that are shed and disseminated through the circulation from the primary tumor to distant organs (circulating tumor cells, CTCs). Although the detection of CTCs may have important prognostic and therapeutic implications, their detection poses key technical challenges because their numbers can be very small. Great efforts have therefore been made toward the enhancement of signal amplification to significantly increase the sensitivity for the detection of tumor-specific biomarkers on the surface of CTCs. In this presentation, I will also present a dual signal amplification immunosensing strategy that has the potential to offer high sensitivity and specificity for the detection of low-abundance tumor cells. High sensitivity is achieved by using graphene to modify the immunosensor surface to accelerate electron transfer and quantum dot (QD)-coated silica nanoparticles as tracing tags. On the other hand, high specificity is obtained by the simultaneous measurement of multiple HCC-specific biomarkers, identified through the microarray studies, on the cell surface of CTCs using different QD-coated silica nanoparticle tracers. In summary, the presentation will cover some of our effort to identify novel biomarkers to diagnose early HCC and the development of novel strategies to couple some of these biomarkers to enable the detection of low abundance circulating tumro cells.






Biography:
Youhe Gao received his MD from Peking Union Medical College, his Ph.D. from University of Connecticut and postdoctoral training from Beth Israel Deaconess Medical Center Harvard Medical School. He is the Professor of Department of Physiology and Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/Peking Union Medical College. His research interests include biomarker discovery in urine proteome, protein interaction and related bioinformatics.
Abstract:
All cells in the body rely on a homeostatic microenvironment to survive and function. Blood, as the key provider of this internal environment for all tissues and organs, is naturally responsible to remain stable and balanced so as to protect organs from exposing to disturbing factors. In contrast, as a filtrate of the blood, urine bears no need or mechanism to be stable. Any change that is introduced into the blood either internally or externally tends to be cleared by the liver, kidney and/or other organs via a variety of mechanisms in order to maintain the homeostasis of the blood. In contrast, urine is the place that most of the wastes in blood are dumped into, and thus tolerates changes to a much higher degree. Biomarkers are the measurable changes associated with a physiological or pathophysiological process. Therefore, they are more likely to be detected in urine. We introduced anticoagulant to disturb the coagulation function of the blood to see if more sensitive changes can be observed in urine than in blood. By using the same proteomic strategy to analyze both urine and plasma samples before and after anticoagulants administration, in heparin group 27 proteins changed in urine but only three in plasma. In argatroban group, 61 proteins were changed but only one of which changed in plasma. This result indicated that even when the functional change was made in the blood itself, change of proteins in urine is still more sensitive to be detected than that in plasma.






Biography:
Wancai Yang was trained as a pathologist in China and received postdoctoral training in Rockefeller University, USA. He is a Professor of pathology and Dean of the School of Basic Medical Sciences at Xinxiang Medical University, China, and an Adjunct Associate Professor of University of Illinois at Chicago, Chicago, Illinois, USA. He has published more than 60 papers in high-impact journals about his research on colorectal and esophageal cancers. He has also been serving as grant reviewer, article reviewer and editorial board members of reputed journals.
Abstract:
Selenium is a trace element that plays a critical role in physiological processes and cancer prevention, whose functions may be through its effects on selenium-containing proteins. One of these, selenium-binding protein 1 (SBP1), is a protein of unknown function that has been shown to be significantly reduced in tumors of diverse tissue types as compared to the corresponding normal tissue. More importantly, SBP1 has also been reported to be a predictor of clinical outcome, higher levels of SBP1 are linked to better prognosis and longer survival time in colorectal cancers. Moreover, the levels of SBP1 are inversely associated with the levels of glutathione peroxidase1 (GPx-1), a protein representative of a different class of selenoproteins, in colorectal and prostate normal and cancer tissues. GPx-1 is an anti-oxidant, selenocysteine containing enzyme implicated in several diseases, including cancer, due to the association of specific alleles with disease risk. Interestingly, increased expression of SBP1 does not alter GPx-1 expression but suppresses GPx-1 enzyme activities, and increased expression of GPx-1 downregulates SBP1 expression levels. The relationship between SBP1 and GPx-1 represents a unique example of a molecular interaction between selenium-containing proteins with a likely significant impact on human health and disease. Taken together, the crosstalk between these two distinct classes of selenium-containing proteins plays a critical role in carcinogenesis and SBP1 could be used as a potential biomarker for cancer progression, prognosis and prognostic indicator of cancer outcomes.






Taly Valerie

Centre Universitaire des Saints, France

Title: Tiny droplets to detect cancer biomarkers
Biography:
Taly Valerie conducts her research at the interface between chemistry, physics and biology. Since 2003, she has been working with Prof. Andrew Griffiths, first in the Medical Research Council in Cambridge (UK) and then in the ISIS (Strasbourg). She focuses her research on in vitro compartmentalization of biological and chemical reactions in emulsion droplets of few picoliters. She recently started the Translational Research And Microfluidics Group within the clinical oncology research unit headed by Prof. P. Laurent-Puig. Her research is dedicated to the clinical validation of droplet-based microfluidics for the non-invasive detection of Cancer biomarkers, the highlighting of new Cancer Biomarkers and the development of original tools and procedures for their detection with applications in personalized medicine, cancer recurrence detection and cancer diagnostics.
Abstract:
Droplet-based microfluidic has led to the development of highly powerful systems that represent a new paradigm in High-Throughput Screening where individual assays are compartmentalized within microdroplet microreactors. By combining a decrease of assay volume and an increase of throughput, this technology goes beyond the capacities of conventional screening systems. Droplets (in the pL to nL range) are produced as independent microreactors that can be further actuated and analyzed at rates of the order of 1000 droplets per seconds. Added to the flexibility and versatibility of platform designs, such progress in sub-nanoliter droplet manipulation allows for a level of control that was hitherto impossible. We will show how by combining microfluidic systems and clinical advances in molecular diagnostic we have developed an original method to perform millions of single molecule PCR in parallel to detect and quantify a minority of mutant sequences within a high quantity of non-mutated sequences in complex mixture of DNA with a sensitivity unreachable by conventional procedures. Finally, to demonstrate the pertinence of our procedures to overcome the clinical oncology challenges, the results of 2 studies will be presented. When the first one addresses our ability to detect minoritary subclones in colorectal tumors and to understand the impact of these subclones on responses and survival of the patients treated by anti-EGFR therapies, the second one aims at demonstrated the possibility to circulating tumor DNA in plasma of patients with advanced colorectal cancers.






Biography:
Following graduation and 10 years at the research laboratory bench at the CRUK's Birmingham University's laboratories and a UK-based diagnostics company, Tony Bartlett has been engaged for 20 years in the early stages of product development and commercialisation of a number of game-changing analytical technology platforms including, but not limited to: the electro-chemiluminescence-based immunoassay platforms now owned by Roche Diagnostics, Affymetrix' GeneChip microarray platform, and a number of other platforms developed by start-up companies. He has held a number of senior positions including VP and General Manager positions in mature life science tools companies before joining SomaLogic to expand the company's footprint into Europe for whom he is the Director of European Commercial Operations & Collaborations. He describes himself as a "technology geek who is still a scientist at heart".
Abstract:
SomaLogic presents a transformative proteomic biomarker discovery and diagnostic application technology that measures 1 to ~1000 human proteins in low sample volumes (~15-75 μL of serum/plasma, tissue homogenate) with a high-performance, high- throughput, and cost-effective assay. This technology is enabled by a new class of DNA aptamers – “SOMAmers™” – that contain novel chemically-modified nucleotides, which greatly expand the physicochemical diversity of the large combinatorial SELEX libraries from which they are selected. We will describe the highly scalable process that we use routinely to monitor changes in subjects or measure differences between clinical samples, taking into account pre-analytical variability, to derive biomarkers for use in differential diagnosis, determination of disease status, and for monitoring or predicting drug-response.






Retnagowri Rajandram

University of Malaya Cancer Research Institute, Malaysia

Title: Serum TRAF-1 as a biomarker for developing clear cell renal cell carcinoma
Biography:
Retnagowri Rajandram completed her B.Sc (Hon I) then her Ph.D. with University of Queensland Australia in 2009 at age 25. She is currently a Senior Lecturer with University of Malaya in Malaysia where she is from. She is an upcoming researcher and is keen in cancer research specifi cally kidney cancer from her honours year. Her broad interest in this fi eld encompasses the investigation of a possible panel of biomarkers for the disease. Her skills comprise of cell culture work including transfections, western immunblotting, RNA and tissue microarray, immunohistochemistry as well as using Aperio image software. Her current research consist of investigating tumour necrosis factor associated factor 1’s (TRAF1) role in renal cell carcinoma (RCC) using human paraffi n embedded tissue and blood from RCC subjects. Her other projects in this area include further defi ning the molecular pathway of TRAF1 in renal cell carcinoma. She has presented her work at local and international conferences, ever since 2005.
Abstract:
Background and Aims: Renal cell carcinoma (RCC) serum biomarkers, that effectively identify the developing cancer non-invasively, are urgently needed. Tumour necrosis factor receptor-associated factor-1 (TRAF-1) is strongly expressed in the proximal tubular epithelium of normal kidney. The proximal tubular epithelium is thought to be the tissue of origin of clear cell RCC (ccRCC), the most common of the RCC subtypes. The TRAF-1 signalling pathway is necessary for immune response and apoptosis regulation. We have found TRAF-1 is significantly decreased in ccRCC, but no serum analyses had been carried out. The aim of this study was to compare tissue TRAF-1 levels with serum levels in control and ccRCC patients from the University of Malaya Medical Centre (UMMC).
Methods: Immunohistochemistry with automated batch staining, and Aperio ImageScope morphometry were used to compare TRAF-1 in 69 ccRCC patients from UMMC with paired normal kidney tissue from those patients. Serum from 15 ccRCC and 15 healthy people were tested for TRAF-1 (ELISA). ANOVA with Tukey's post-hoc (tissue) and Mann-Whitney U-test (serum) was used (mean ±SEM).
Results: Compared with normal kidney (168512 ± 6166 positive pixel counts/PPC), ccRCC was lower (82591 ± 5646 PPC) (p<0.05). However, TRAF-1 in serum from ccRCC patients was significantly increased over normal serum levels (202.28 ± 74.58 vs 50.63 ± 13.56 ng/ml; p=0.012).
Conclusion: To the best of our knowledge, this is the first study to evaluate serum TRAF-1 levels in patients with ccRCC. Serum TRAF-1 levels were significantly elevated in patients with ccRCC compared with healthy individuals. The increased serum TRAF-1, compared with decreased tissue TRAF-1 in ccRCC, may indicate the protein is actively secreted from developing ccRCC. As such, serum TRAF-1 may be a useful addition to biomarker panels.






Biography:
Gulafshana Hafeez Khan did her medical degree from University of Peshawar, Pakistan and then did post graduation in Obstetrics and Gynaecology from College of Physicians and Surgeons Pakistan. She has trained in Obstetrics and Gynaecology in the UK and is a Member of Royal College of Obstetricians and Gynaecologists since 2004. She is currently doing PhD in Obstetrics and Gynaecology in School of Medicine, University Of Nottingham. Her special interest is reproductive medicine and general Obstetrics.
Abstract:
Background: Various studies have indicated an association between Polycystic Ovary Syndrome (PCOS) and pre-eclampsia (PE), however the pathophysiological mechanisms supporting this association are unclear. Defining such mechanisms linking PCOS and PE, could assist in more accurate screening, and underpin novel preventative strategies for pre eclampsia. Aim: The aim of this study was to catalogue proteomic biomarkers in PE and considers whether any represent candidate biomarkers for the detection of PET risk in women with PCOS. Methods: A systematic review and update of our PCOS proteomic biomarker database was performed, along with a parallel review of PE biomarkers. All eligible published studies on proteomic biomarkers for PE and PCOS which were identified through MEDLINE (1996- December 2013), EMBASE (1980- December 2013) and Cochrane (1993- December 2013) ISI web of knowledge (v4.2) databases, using the search terms “proteomics”, “proteomic”, “pre-eclampsia”, “pre-eclamptic toxemia, “proteomic biomarker”, “proteomic biomarker” and “polycystic ovary syndrome” without any restrictions were evaluated. The primary studies on PE were used to create a first database of proteomic biomarkers for PE, whereas those on PCOS to update our existing PCOS database. The two databases were then compared and searched for overlaps; proteomic biomarkers that were common to women with PCOS and PE. Results: Five proteomic biomarkers were differentially expressed in both in women with PE and PCOS compared to controls: transferin, fibrinogen alpha, beta and gamma chain variants, kininogen-1, annexin 2 and peroxiredoxin 2. In PE the biomarkers were found in serum, plasma and placenta and in PCOS the biomarkers were identified in serum, follicular fluid, ovarian and omental biopsies. Conclusions: These proteomic biomarkers could potentially be used to understand the pathophysiological mechanisms linking PCOS and PE. The biomarkers may help to identify subgroups of women with PCOS at risk of developing PE and early diagnosis of PE could be helpful in the management of the women at risk.






Symposium on
Session Chair

Wayne Carter

University of Nottingham, UK




Session Introduction
Biography:
Wayne Grant Carter received his Honours degree in Biochemistry with Nutrition from the University of Southampton. He then completed a Ph.D. at the University of Southampton studying the molecular signalling cascade elicited by insulin (Carter et al., 1995, Asamoah et al., 1995; Carter et al., 1996) supervised by Dr. Graham Sale. Dr. Carter then moved to the Babraham Institute, Cambridge, to work with Professor Jeremy Saklatvala, on studies to elucidate the signalling mechanisms activated as a response to pro-inflammatory cytokines (Finch et al., 2001). Dr. Carter then relocated with Professor Saklatvala's group to the Kennedy Institute of Rheumatology, part of Imperial College, London. Subsequently, Dr. Carter moved to the University of California at Irvine to work with Professor Dana Aswad, similarly studying protein post-translational modification, and protein damage and repair (Young et al., 2001). Dr. Carter then joined the Department of Human Anatomy & Genetics headed by Professor Kay Davies CBE FRS at the University of Oxford, before moving to take up an industrial post with Mobious Genomics, Exeter. He is currently an Editorial Board member of the BIOINFO Journal of Proteomics; Current Chemical Research; Journal of Analytical Sciences, Methods and Instrumentation; World Journal of Biological Chemistry; and Executive Editor for Europe for Journal of Chromatography and Separation Techniques.
Abstract:
A biological biomarker may provide a valuable means todetect, measure, or quantify normal biological activity or that associated with a disease, or assess the effects of a disease treatment. Differential proteome profiling is often used to provide a basis for mining suitable biomarkers of disease. However, patho-physiological changes may be manifested as either a difference in protein post-translational modification (PTM) levels and/or alteration in protein associations. These processes may arise in the absence of protein level changes, and may therefore be missed by conventional biomarker proteomics. We have assessed proteinPTM in the cerebrospinal fluid of multiple sclerosis (MS) patients and controlsubjects. We report the detection of novel protein biomarkers in MS patients via radiolabelling of PTMs. The sensitivity afforded by protein PTM radiolabelling and autoradiography provided a basis to detect novel protein biomarkers, and characterise the proteins by one dimensional and two dimensional separation techniques. This sensitivity and detection of protein PTM provided a viable means to track these novel biomarkers, and enable purification to a level sufficient for mass spectrometry identification






Track 2: Functional Genomics and Cytogenetic Biomarkers
Session Chair

Marija Krstic-Demonacos

University of Manchester, UK




Session Co-Chair

Carlo Emanuele Neumaier

Universitaria San Martino-IST, Italy




Session Introduction
Biography:
Carlo Emanuele Neumaier studied Medicine and Surgery at University of Genoa (Italy) and specialised in Radiodiagnostic at the University of Genoa. He is currently a Contract Professor at the Department of Radiology and Oncology at the University of Genoa and he works at the Diagnostic Imaging and Senology Unit, IRCCS– Azienda Ospedaliera Universitaria San Martino-IST-National Cancer Institute, Genoa. He has a strong experience in MRI, CT, Ultrasound, and Optical Imaging and he has written several scientific papers in peer-reviewed journals and book chapters. He is Principal Investigator of several national clinical research projects about the study of prostate cancer and breast cancer by Magnetic Resonance. He is invited speaker to national and international conferences and he is an active member of the European Society of Molecular Imaging (ESMI).
Abstract:
Prostate cancer (PCa) is the most frequently diagnosed malignancy in men. Actually, prostate-specific antigen (PSA) and digital rectal examination (DRE), are the most accepted tools for screening. PSA is known to be prostate specific, but not PCa specific. It is controversial the appropriate level of serum PSA that triggers an invasive approach by a template TRUS-guided biopsy. This technique lacks in sensitivity with a 30-45% risk of “up or down-staging” of PCa. The recent development in imaging field, suggest the emerging role of the multi-parametric Magnetic Resonance Imaging (mpMRI) as tool for the diagnosis of PCa. MpMRI consist in the combination of functional study, such as, diffusion-weighted imaging (DWI), dynamic contrast enhanced imaging (DCE-MRI), MR spectroscopy (MRS), and the morphological, by T2-weigheted imaging. This technique, provides crucial information regarding tumour location, volume, grade and extension. Thus mpMRI can find a possible role to guide biopsies, in order to improve the identification of aggressive tumor. Furthermore, in the case of low grade tumor, this technique, represents an optimal tool for the active surveillance. Here, we highlight, the mpMRI as “imaging biomarker” in the diagnosis, staging, grading and treatment planning of prostate cancer and we discuss its alternative role to TRUS-guided biopsy.






Biography:
Raif Yuecel the Head of the Iain Fraser Cytometry Centre at the University of Aberdeen, servicing and consulting in biology, biochemistry, and medical research division labs on Cytometry, and also on Small Animal Imaging Applications. His research is mainly focused on different application possibilities of Cytometry, such as the investigation of cellular signalling in human blood, stem cell analysis, cancer research, biomarker discovery, immunology, clinical research, microbiology cytometry, marine and ocean biology. As principal investigator he is also actively involved in teaching and consulting on Cytometry. Dr. Raif Yuecel is a member of various Cytometry and Imaging society (ISAC, FCUK, DGfZ, WMIS, ISMRM).
Abstract:
Research of cellsignalling pathways, networks, and interactions that cells require to function is a major scientific challenge. Kinases and Signal Transducers and Activator of Transcription (STAT) proteins regulate many aspects of cell growth, survival and differentiation. The transcription factors of this family are activated by kinases (e.g. JAK) and dysregulation of this pathway is frequently observed in primary tumors and leads to increased angiogenesis, enhanced survival of tumors and immunosuppression. Usually, cell signalling analysis has been performed using Western blots on large cell populations. If performed on a complex combination of cells, such as peripheral blood, this technique would not yield signalling information of individual cellular subsets within the whole population. Intracellular flow cytometric analysis of protein phosphorylation has emerged as a powerful tool in the field of immunological signalling, which allows to analyze quickly and accurately the cellular subsets in heterogenous populations. Here we are describing the evaluation of multiparametric flow cytometry application to investigate the early phosphorylation events of STAT3, STAT5 and STAT6 in human peripheral blood. For this purpose we evaluated the brief application of cytokines and mitogens in vitro and analyzed the phosphorylation of these STAT proteins in the human leukemic monocyte lymphoma cell line U937. Individual cytokine combination(IL2, IL4, IL6 and GMCSF) of treatment were assessed to ensurethe detection of specific STAT phosphorylation .In conclusion, detection of STAT3, STAT5 and STAT6 phosphorylation through intracellular staining by Flow Cytometryin multiparametric analysis enables the study of cell signaling events in an approach that is quick, sensitive and quantitative on the single cellular level compared to conventional methods.






Biography:
Alessio Naccarati has completed his Ph.D. in Human Molecular Genetics in 2003 at Pisa University (Italy). He has made his postdoc in the Department of Molecular Biology of Cancer (Academy of Sciences of the Czech Republic, Prague), where has also held a position of Head of Laboratory. He is currently Senior Scientist at the Molecular Genetic and Epidemiology Unit at Human Genetics Foundation (Torino, Italy). He has published 68 papers in peer reviewed impacted journals, participated in several international/national grants, both in Italy and Czech Republic. His main topics are colorectal cancer susceptibility, DNA repair and recently microRNAs.
Abstract:
MicroRNAs (miRNAs) are key gene regulators in most biological and pathological processes, including colorectal cancer (CRC). The association of miRNA-related polymorphisms with CRC incidence and prognosis, as well as the possibility of using circulating or fecal miRNA expression as noninvasive early detection biomarkers open interesting possibilities. miRNAs may be potential molecular classifiers, early detection biomarkers, and therapeutic targets for CRC. However, the underlying mechanisms of aberrant miRNA regulation, such as altered DNA methylation, are still poorly known. We report our investigations on a) the role of variation in miRNA target binding sites on CRC risk and prognosis, focusing on DNA repair genes, b) genome-wide miRNA methylation levels in prediagnostic samples, c) the search of CRC biomarkers in surrogate specimens. Variations in predicted binding sites in 3’UTR of nucleotide-excision repair pathway genes were associated with CRC risk, while a polymorphism in base-excision repair gene SMUG1 showing reduced gene expression in vitro was associated with increased survival in patients treated with 5-fluorouracil-based therapy. A miRNA signature discriminating CRC and precancerous lesions has been described in plasma samples, showing a dysregulation of miR-34a and miR-150. A pilot study on stool/plasma from healthy volunteers with different dietary habits has revealed a differential expression of miR-92a among vegans, and subjects with omnivorous diet. A novel approach to analyse miRNA methylation from epigenome-wide DNA methylation levels measured in peripheral blood samples is described. The necessity to move on high-throughput techniques to globally define miRNA signatures involved in colorectal carcinogenesis in surrogate specimens is also briefly discussed.






Biography:
McMaster received a B.Sc and M.Sc from the University of BC was awarded a D.Phil. from the University of Oxford. He joined the UBC Department of Medical Genetics in 1982, and served as Head of the UBC Department of Medical Genetics and Head of Medical Genetics for the Women’s Health Centre for British Columbia, 2000-2010. Dr. McMaster was also Founding Director of the Immunity and Infection Research Centre at VCH Research Institute and Director of Transplant Immunology for British Columbia Transplant until being appointed in 2008 as Vice President, Research for VCH, Executive Director, VCH Research Institute, and Associate Dean of Research, Faculty of Medicine, UBC.
Abstract:
Solid organ transplantation is often the only treatment for end stage organ failure and has shown a dramatic increase in outcomes over the past 20 years. However, acute graft rejection remains an important clinical concern and is an adverse predictor for long-term graft survival. The current gold standard diagnostic for acute rejections remains repeated tissue biopsies–a highly invasive procedures with associated risks. Using advanced genomics, proteomics and computational biology, a series of blood based diagnostic biomarkers for acute heart allograft rejection have been developed and are currently being clinically implemented to monitor timely and effective therapeutic intervention to minimize graft damage and enable knowledgeable adjustment of immunosuppressive therapy. Using a series of blood samples from over 65 transplant recipients with or without acute rejection, panels biomarkers correlating with acute rejection were generated using leukocyte mRNA and whole genome microarrays and quantitative plasma proteomics. The biomarkers were then validated using a separate series of 85 single time point samples from a cross-Canada cohort of recipients. The resulting validated panel of 10 genes exhibit excellent performance with 100% sensitivity and 74% specificity and 0.85 AUC (100% NPV and 14% PPV), that increases to 100% sensitivity and 91% specificity and 0.91 AUC (100% NPV and 32% PPV) with the addition of 6 plasma proteins biomarkers. Multiplex assays for both genomic and proteomic biomarkers are currently being developed for FDA approval for implementation in clinical laboratories and routine transplant monitoring.






Biography:
Laszlo Takacs is the Founder and CSO of Biosystems International, Professor, Department of Human Genetics at Medical Center of the University of Debrecen, Hungary, Foreign Member of the Hungarian Academy of Sciences. He received his education at Semmelweis Medical University in Budapest. Experience includes over 10 years of academic research in Hungary and at the NCI, NIH in the US and 20 years in industry at Amgen Inc. and Pfizer Inc. In 2004, Laszlo Takacs co-founded Biosystems International. Publications include over 70 peer reviewed scientific papers, book chapters and commercially oriented materials. Laszlo Takacs is an inventor on 10 patents and patent applications. His current interest is human plasma protein epitome profiling for the discovery of new diagnostics in cancer and chronic diseases via mAb proteomics.
Abstract:
The ultimate biological function is associated with proteins not nucleic acids, thus there is considerable interest in understanding proteome variability and its association with disease. Genetic code dependent proteome variability (splice variation, coding SNPs, etc.) is only a part of proteome variability, which includes genetic code independent heterogeneity such as post-translational modification, protein interactions and folding variation. We show here that disease diagnosis may benefit from the large scale profiling of proteins and their genetic code dependent and independent variability at the level of protein epitopes in the human plasma. Monoclonal antibody targeted detection and quantification of the multitude of protein variability is feasible via epitope profiling, in contrast to MS based or other affinity reagent based technologies which fail to detect genetic code dependent and independent variability. Epitope specific non-redundant mAb libraries running on suitable antibody biochip platforms, such as the Randox Evidence Investigator platform, provide diagnostic level combinatorial markers. We present results of human plasma proteome profiling experiments from observational studies which indicate an unprecedented >95% accuracy (AUC in ROC analysis) for the detection of lung, breast, colon and prostate cancers outperforming current multiplexed or individual markers. Our current focus includes bringing the technology to closer to patients and to the diagnostic market.






Biography:
After 2 years of researcher position in the Anticancer centre in Montpellier, Commes Therese was recruited as Assistant Professor (1992-2007) and as Professor in 2007 at the University Montpellier 2. Head of the group “Transcriptomics, bioinformatics and myeloid leukemias” at INSERM U1040-Montpellier, she is involved in interdisciplinary projects since 2005. Since 2012, She is involved in the advisory board of the “computationnal Biology Institut (IBC)” (Director: O Gascuel) and in the management of the program “Methods for high‐throughput sequencing analysis” with E Rivals (http://www.ibc-montpellier.fr/wp/wp1), Advisory board of the Montpellier-Genomic platform (MGX) and the Bioinformatics platform (ATGC, LIRMM, ReNaBi), member of the FRANCE GENOMIQUE Bioinformatics network (WP“RNA-seq”). Contribution to the creation and Consultant of the Skud-tech company.
Abstract:
The comprehensive analysis of expression profiles based on RNA-seq provides unprecedented sensitivity for exploring transcriptome in all its complexity. The difficulty lies in the ability to detect and extract rigorously this type of information before biological validations. To this end, we have developed a new program called Crac, more efficient than the tools currently used in the field and which is based on an innovative algorithm and on double indexing the human genome and RNA-Seq data (crac.gforge.inria.fr/: Philippe et al, 2013). It allows extracting with high specificity and sensitivity, all the biological transcriptional events irrespective of annotation (substitutions, indels, splice junctions, potential chimeric RNAs…). The strong point of our approach is the ability to use our own program Crac for categorizing transcripts variants specifically new splice junctions and chimeric RNAs which could have a role in the establishment of tumor mechanisms, and to use a new integrated transcriptome analysis procedure for the characterization and the discovery of new non-coding RNAs (ncRNA) combining data from both RNA-seq and Digital gene expression (DGE) in order to identify transcriptional abnormalities, not only in assessing changes in gene expression, but by analyzing the complete repertoire of transcription. Applications in the study of myeloid leukemia are currently done. Investigating the biological features of novel chimeric RNA and ncRNA candidates with such combinatorial approach is an interesting and challenging goal to achieve as it can pave the way for identifying the new potential prognostic and diagnostic biomarkers.






Track 5: Biomarkers in Clinical Research and Development
Track 7: Biomarkers of Exposure Response and Susceptibility
Session Chair

Sergey Suchkov

I.M.Sechenov First Moscow State Medical University, Russia




Session Co-Chair

Pavel Vodicka

Institute of Experimental Medicine, Czech Republic




Session Introduction
Biography:
Sergey Suchkov graduated from Astrakhan State Medical University and was awarded with MD.\In 1985, Suchkov obtained his Ph.D. As a Ph.D. student of the I.M. Sechenov Moscow Medical Academy and Institute of Medical Enzymology, USSR Academy of Medical Sciences, Moscow, Russia. In 2001, Suchkov finished the PostDoc Research Fellowship Program and maintained his Doctor Degree at the National Institute of Immunology, Russia. From 1987 through 1989, Dr. Suchkov was a senior Researcher, Lab of Developmental Immunology, Koltzov Institute of Developmental Biology, USSR Academy of Sciences to deal to developmental immunology. From 1989 through 1995, Dr. Suchkov was being a Head of the Lab of Clinical Immunology and Im-munobiotechnology, Helmholtz Eye Research Institute in Moscow. From 1995 through 2004, Dr. Suchkov was being a Chairman of the Department for Clinical Immunology, Moscow Clinical Research Institute (MONIKI) and the Immunologist-in-Chief of the Moscow Regional Ministry of Health. At present, Dr Sergey Suchkov, M, Ph.D., is Professor in Immunology, Department of Pathology, School for Pharmacy, I.M. Sechenov First Moscow State Medical University, Dean of the Department (Faculty) of The PPPM Development, and the First Vice-President of the University of World Business, Politics and Law and Secretary General, United Cultural Convention (UCC), Cambridge, UK.
Abstract:
The methodological bricks of subclinical diagnostic protocols should include basic algorithms to differ essentially from those employed in traditional clinical practice, i.e., (i) to confirm a diagnosis of subclinical stage of the disease course and (ii) to select a mode for preventive treatment to quench the autoimmune inflammation. In this sense, among the best-validated proteome-related biomarkers, antibodies (Abs) are the best known ones to represent one of the principal immune effectors and thus key mediators of inflammatory responses to generate the events. Most of autoimmune disorders including multiple sclerosis (MS) are preceded by a symptom-free subclinical stage in which the patients can be identified by specific autoAbs. The sequence-specificity of Ab-proteases demonstrates five sites of preferential proteolysis to be located within the immunodominant regions of MBP confirmed by the structural databanks. Cleavage at those sites occurred at a similar rate as determined by 32P-MBP degradation assay. Those sites are located within the immunodominant regions of MBP; and two of them falling inside the sequence covering a 81-103 peptide segment and its 82-98 subsegment as well, with the highest encephalitogenic properties both to act as a specific inducer of EAE in SJL mice and to be attacked by the MBP-targeted Ab-proteases very often in MS patients with the most severe (progradient) clinical courses. Meanwhile, sites localized within the frame of 43-68 and 146-170 peptide subsegments whilst being less immunogenic happened to be EAE inducers very rare but were shown to be attacked by Ab-proteases very often in MS patients with moderate (remission-type) clinical courses. To test the Ab-proteses specificity toward distinct MBP fragments, recombinant fusion proteins of Trx with the C-terminally fused MBP peptides were designed, and in all cases, recombinant substrates were cleaved only at preferential sites inside the MBP fragment, leaving the Trx part undegraded. These data further confirmed the substrate specificity of the Ab and its profound difference from trypsin, a common protease that cleaves at basic residues. Finally, in moderate (remission-type) courses, Ab-proteases focuse its proteolytic effect on low-immunogenic and low-encephalitogenic 43-68 и 146-170 sites but in aggressive cases (progradient courses), the Ab-mediated proteolysis was prevailed on highly-immunogenic and highly-encephalitogenic 81-103 and 82-98 sites. And registration in the evolution of highly immunogenic Ab-proteases to attack 81-103 and 82-98 sites predominantly would illustrate either risks of transformation of subclinical stahes into clinical ones, or risks of exacerbations to develop. The traditional applications of assays for canonical antimyeline Abs for diagnostic and prognostic purposes in patients with a clinically isolated syndrome (CIS), a frequent precursor to clinically definite MS (CDMS), has yielded conflicting results. Meanwhile, the activity of Ab-proteases in combination with the sequence-specificity would confirm a high subclinical and predictive value of the tools as applicable for personalized monitoring protocols. It is so important to stress that the close association between the proteolytic sensitivity of MBP and post-translational modifications of the latter may represent one of the key regulatory mechanisms in the epitope generation. For sure, a combinative (enzyme- and Ab-mediated) proteolysis may illustrate a crucial pathway to exert a concerted attack on MBP, although mechanisms responsible for the activation of these potential activities are not known yet. Ab-proteases can be programmed and re-programmed to suit the needs of the body metabolism or could be designed for the development of principally new catalysts with no natural counterparts. Of tremendous value are Ab-proteases directly affecting the physiologic remodeling of tissues with multilevel architectonics (for instance, myelin). By changing sequence specificity of the Ab-mediated proteolysis one may reach reduction of a density of points of the negative proteolytic effects within the myelin sheath and minimizing scales of demyelination. And, autoAb-mediated proteolysis could thus be applied to isolate from Ig molecules the efficient catalytic domains directed against particular autoimmune epitopes pathogenically and clinically relevant (encephalitogenic epitopes). Further studies on targeted Ab-mediated proteolysis may provide a supplementary tool for predicting demyelination and thus the disability of the MS patients.






Biography:
Pavel Vodicka graduated at the Medical Faculty, Charles University, Prague and in 1986 obtained PhD in biochemistry. He worked as postdoctoral fellow at the Finnish Inst. Occupat. Health, Helsinki, Finland (1987-1990) and as visiting scientist at Karolinska Institute, Huddinge, Sweden (1990-1993). Since 2002 he heads the Dept. Molec. Biol. Cancer, Inst. Exper. Medicine, Acad. Sci., Prague, Czech Republic. Pavel Vodicka has published more than130 (total IF 552.4, 2650 citations a HI 31) articles. Since 2004, his main research topics are focused on the DNA and chromosomal damage and DNA repair functional tests in humans and on transient biomarkers in the onset of gastrointestinal cancers. In 2012 he edited the Special Issue in Mutagenesis (http://mutage.oxfordjournals.org/content/27/2.toc), entitled Colorectal Cancer-Current Insights into Susceptibility.
Abstract:
Background: Majority of cancer cases occurs sporadically with an important involvement of genetic variants in low penetrance genes. Sporadic cancers are additionally hallmarked by a complex interplay between the genetic and environmental factors. Colorectal carcinogenesis (CRC), as a complex process, involves a plethora of events resulting in genomic instability. The theories underlying carcinogenic process point out either the role of somatic mutation or the surrounding microenvironment, however neither of them explains all features of cancer. Uncontrolled proliferation and genomic instability point to the DNA repair and DNA damage response as to the key players. DNA repair protects the integrity of the genome and ensures the general and specialized functions of cells, thus preventing the carcinogenesis. Consequently, differences in DNA repair efficiency pose an important issue in sporadic CRC etiology, affected by complex gene-environment (microenvironment) interactions. In the present study, we will overview several biomarkers in mapping heterogenous complex CRC disease, which poses a serious health problem in the Czech Republic. Finally, we will provide an excursion into the DNA repair phenotypic reflection in both the control individuals and in CRC patients.
Materials and Methods: Gene variants in candidate genes were analyzed on highthroughput platforms by using Real Time PCR instruments (Taqman allelic discrimination, Polakova et al. 2009), by High Resolution Melting analysis and also NGS. Relevant mutations in candidate genes will be assayed by HRM analysis (LightCycler®480, Roche), denaturating capillary-electrophoresis (ABI PRISM 310 System, Applied Biosystems) and next generation sequencing (GS Junior, Roche). The cDNA and cRNA arrays for determination of mRNA, miRNA and lncRNA transcript levels and following bioinformatics analysis were carried out by qRT-PCR (ABI7500, Applied Biosystems). Determination of methylation level in promoter sequences of both microRNA and protein coding genes was assayed for by using high resolution melting analysis following bisulfite conversion. Imunohistochemical analysis (IHC) for protein expression was assayed for all the genes, picked up as relevant ones in the course of follow-up. Western blottings (WB) were deteremined in candidate proteins (such as DNA repair protein hOGG1) by using the specific antibodies. The analyzis of functional aspects of DNA repair has recently been published (Slyskova et al. Clin. Cancer Res. 2012).
Results: Variants in genes involved in several important pathways, such as DNA repair, cell cycle control, folate metabolism and methylation, insulin resistance and obesity, ABC transporters, selenoprotein genes, genes involved in inflammatory/immune response have shown various degree of association with CRC risk. We hereby present the data on mutations in high risk genes involved in colorectal carcinogenesis. Gene expression levels were determined in relevant pathways and complemented with other important parameters, such as epigenetic regulators of transcription by methylation. Additionally, the role of post-transcriptional regulation via miRNA or lncRNA was investigated in relation to the risk of CRC and the efficacy of chemotherapy. Our experiments revealed SNPs in miRNA binding sites of SMUG1 gene affect significantly survival in 5-fluorouracil-treated CRC patients. For instance, polymorphisms in miRNA binding proved the study hypothesis and highlighted the importance of SNPs within miRNA-dependent regulatory regions. Within important candidate pathways we assayed for the expression of the specific proteins (mismatch repair, base excision repair). As an additional marker of individual susceptibility to sporadic CRC and potential prognostic predictor functional DNA repair tests have been implemented as biomarkers of the complex phenotype.
Conclusions: An application of the whole set of various biomarkers, covering genetic, epigenetic and functional aspects contribute to define the phenotypic landscape of the disease and, in some instances, to delineate the individual response to the therapy. Moreover, DNA repair capacity measurements (functional DNA repair assays) represent a complex marker integrating genetic polymorphisms, gene expression, stability of gene product, effect of inhibitors/stimulators, environmental factors and lifestyle factors. Functional DNA repair assays reflect the capacity of the organism to cope with a chronic exposure to numerous environmental and dietary genotoxicants and may also be used as predictive markers in cancer therapy.






Ondrej Topolcan

Charles University in Prague, Czech Republic

Title: Clinical usefulness of HE4 in endometrial cancer diagnostics
Biography:

Abstract:
Problem statement: Our aims were to evaluate the benefits of the determination of human epididymis protein 4 (HE4) in the endometrial cancer diagnostics. We compared HE4 with cancer antigen 125 (CA125).
Methods: The total number of females was 66. The patient group consisted of 32 females with endometrial cancer and the control group of 34 healthy females. All diagnoses were verified histologically with equal representation of tumors in FIGO stage I - IV. Age distribution of Ca endomerial group and healthy controls group were very similar. The average age was 58.5 and 65 years. HE4 were measured using ELISA kit (Fujirebio Diagnostics, Sweden). CA125 were measured using DxI instrument (Beckman Coulter, USA). The SAS 9.2 Software was used for all statistical analysis. Wilcoxon test was used to the comparison of cancer and healthy group.
Results: When HE4 was evaluated at 96.88% specificity: cut-off: 90.0 pmol/L, sensitivity: 41.18%, positive predictive value: 93.33%, negative predictive value: 60.78% and an area under the curve: 0.8056. Comparison of distribution of Wilcoxon score between cancer and healthy group was statistically significant p<0.0001. CA125 was evaluated at 96.88% specificity: cut-off: 42.0 IU/L, sensitivity: 20.59%, positive predictive value: 87.50%, negative predictive value: 53.45% and an area under the curve: 0.5700. Comparison of distribution of Wilcoxon score between cancer and healthy group was statistically not significant p=0.4442.
Conclusions: Measurement of HE4 is more sensitive than CA125 determination. Determination of HE4 is an useful tool for improving of endometrial cancer detection and next strategy of treatment.






Biography:
Wayne Grant Carter received his Honours degree in Biochemistry with Nutrition from the University of Southampton. He then completed a Ph.D. at the University of Southampton studying the molecular signalling cascade elicited by insulin (Carter et al., 1995, Asamoah et al., 1995; Carter et al., 1996) supervised by Dr. Graham Sale. Dr. Carter then moved to the Babraham Institute, Cambridge, to work with Professor Jeremy Saklatvala, on studies to elucidate the signalling mechanisms activated as a response to pro-inflammatory cytokines (Finch et al., 2001). Dr. Carter then relocated with Professor Saklatvala's group to the Kennedy Institute of Rheumatology, part of Imperial College, London. Subsequently, Dr. Carter moved to the University of California at Irvine to work with Professor Dana Aswad, similarly studying protein post-translational modification, and protein damage and repair (Young et al., 2001). Dr. Carter then joined the Department of Human Anatomy & Genetics headed by Professor Kay Davies CBE FRS at the University of Oxford, before moving to take up an industrial post with Mobious Genomics, Exeter. He is currently an Editorial Board member of the BIOINFO Journal of Proteomics; Current Chemical Research; Journal of Analytical Sciences, Methods and Instrumentation; World Journal of Biological Chemistry; and Executive Editor for Europe for Journal of Chromatography and Separation Techniques.
Abstract:
Undesired exposure to xenobiotic compounds such as commercial or domestic pesticides is a major health concern. Some of the most widely employed pesticides are organophosphorus (OP) compounds that, similar to nerve agents, adduct and inhibit acetylcholinesterase (AChE) within nerve synapses. OPs also adduct other secondary protein targets for which health consequences are equivocal and incompletely defined. Protein post-translational modification (PTM) by adduction may influence protein properties such as enzymatic activity or immunogenicity. Hence there is a need to identify all secondary targets of pesticides that may be modified by PTM after either acute or cumulative pesticide exposures and the consequences of protein adduction fully determined. Additionally, secondary targets may provide useful biomarkers of pesticide exposure that display superior sensitivity to sole measurements of AChE inhibition. Herein we detail our characterisation and identification of secondary targets of commonly encountered OPs, and discuss their potential to act as both biomarkers of pesticide exposures and decoy surrogate binding targets.






Biography:
Klara Valyi-Nagy, MD is Research Assistant Professor at the Department of Pathology, University of Illinois at Chicago and Associate Director of the University of Illinois Biorepository. Her primary research interests include three-dimensional cultures in oncology research and tissue banking and extracellular matrix - genome interactions in cancer.
Abstract:
Vasculogenic mimicry (VM) patterns are present in numerous malignant tumor types, represent the formation of perfusion pathways by tumor cells, and their presence in tumors is associated with adverse outcome. Mechanisms by which VM may contribute to adverse outcome are not well understood. Previous observations in our laboratory indicated that VM-forming tumor cells in three-dimensional (3D) uveal melanoma cultures have increased resistance against oncolytic virotherapy. To determine whether VM-forming tumor cell subpopulations also have increased resistance against cytotoxic drugs, traditional two-dimensional (2D) and extracellular matrix (Matrigel)-containing 3D cultures of C918 uveal melanoma cells were established and exposed to cisplatin or cadmium chloride. We found that VM-forming tumor cells demonstrated prolonged survival relative to other tumor cell subpopulations in 3D cultures and to cells grown in 2D. To explore the possibility that the increased therapy resistance of VM-forming tumor cells is due to a cancer stem cell phenotype, the expression of cancer stem cell marker CD271 was determined in 2D and 3D uveal melanoma cultures by fluorescent immunocytochemistry. We found that the VM-forming tumor cell subpopulation in 3D cultures expressed CD271. In contrast, cells grown in 2D cultures and tumor cell subpopulations not participating in VM formation in 3D cultures were negative for CD271. These findings suggest that increased drug resistance is a mechanism by which VM-forming tumor cells contribute to adverse outcome. Our findings also suggest that VM-forming uveal melanoma cells acquire a cancer stem cell-like phenotype that may play a role in the increased therapy resistance of these cells.






Biography:
Yingyan Yu graduated from Okayama University, School of Medicine of Japan in 1993 and got M.D., Ph.D. She has got a professional training for diagnostic pathology in UPMC, Pittsburgh University, USA in 2002. Now, she acts as a Professor of molecular pathology in Shanghai Ruijin Hospital, Shanghai Institute of Digestive Surgery, affiliated to Shanghai Jiao Tong University, School of Medicine. She is engaged in translational research on gastrointestinal tumor. The research field of Professor Yu is focused on translational medicine of gastrointestinal cancer, including the relationship of cancer phenotype with the molecular variation, tumor biomarkers for early diagnosis, prognostic prediction and molecular classification for gastrointestinal carcinomas. The goals of her research are to discover the mechanism of heterogeneous histology phenotypes for gastric cancer. For this purpose, molecular genetics, protein and tissue microarray methodologies as well as whole genome sequencing have been used for the studies on human tissue, blood and urine material. Professor Yu has published over 100 papers in English and Chinese.
Abstract:
Gastric cancer is one of the most common malignancies in China. So far, there are few reliable serum biomarkers for diagnosis. The available biomarkers of CEA, CA19-9 and CA72-4 are not sufficiently sensitive and specific for gastric cancer. In this study, a high density antibody microarray was used for identifying new biomarkers from serum samples of gastric cancer. Serum samples from colorectal cancer, pancreatic cancer, hepatocellular cancer and breast cancer were also screened for comparative study. As result, some candidate biomarkers were found out. IPO-38, one of up-regulated serum proteins in gastric cancer was selected for subsequent validation including serum IPO-38 expression by ELISA, IPO-38 protein expression by immunohistochemistry. The immunoprecipitation by IPO-38 for gastric cancer cell line and MALDI–TOF/TOF mass spectrometer suggested that pull-down of IPO-38 belongs to H2B histone, which was supported by co-localization study of laser scanning confocal microscope. Follow-up study showed that survival rate of IPO-38 negative group was better than that in IPO-38 positive group. The study firstly clarified the property of IPO-38 proliferating marker, and proposed that IPO-38 protein is a promising biomarker both for diagnosis and for predicting prognosis of gastric cancer.






Biography:
Evi Lianidou is currently Professor of Analytical Chemistry and Clinical Chemistry at the Department of Chemistry, University of Athens, Greece. Dr. Lianidou is an elected member of the Committee for Clinical Molecular Biology Curriculum of the International Federation of Clinical Chemistry (IFCC), that offers training in Molecular Diagnostics all over the world by the way of courses and hands on workshop in combination with lectures and methodological issues. Dr. Lianidou is coordinating the M.Sc. program of Clinical Chemistry, at the Department of Chemistry, University of Athens and has established a Molecular Diagnostics Laboratory at the Department of Chemistry since 1995. Her lab is specializing in the Analysis of Circulating Tumor Cells, and has access to many patient samples through extensive clinical collaborations, especially with the University of Crete Medical School. Her main research interests are especially on: a) study of micrometastasis through the development of singleplex and multiplex quantitative real time RT-qPCR assays for the detection of Circulating Tumor Cells (CTCs), b) development and clinical evaluation of DNA methylation assays in fresh tissues, paraffin-embedded breast carcinomas, in CTC and in cell-free circulating DNA, c) development and clinical evaluation of quantitative real time RT-qPCR assays for the detection of mature micro RNAs in fresh tissues, paraffin-embedded breast carcinomas and in cell-free circulating DNA, d) development and clinical evaluation of multiplex assays for gene expression in CTCs based on the Luminex bead array system. Dr. Lianidou has 80 publications (http://www.ncbi.nlm.nih.gov/pubmed/?term=lianidou) and has organized: a) the 7th International Symposium on Minimal Residual Disease in Athens, (http://ismrc2009.chem.uoa.gr), b) a scientific meeting on CTCs “Advances in Circulating Tumor Cells: From Basic Research to Clinical Practice” (www.actc2012.org), and c) is currently organizing the next ACTC meeting, on October 8-11th in 2014, in Crete, Greece (www.actc2014.org).
Abstract:
Detection of Circulating Tumor Cells (CTC) in peripheral blood can serve as a "liquid biopsy" approach and has thus emerged lately as one of the hottest fields in cancer research. The clinical significance of CTC has been evaluated in many types of solid cancers, and the CTC enumeration test in metastatic breast, colorectal and prostate cancer has been cleared by the FDA almost a decade ago. CTC molecular characterization has a strong potential to be translated into individualized targeted treatments. A variety of analytical systems are continuously been developed for CTC isolation, detection and molecular characterization. The main strategies are based on their separation from peripheral blood mononuclear cells based on CTC density, size and electric charges and protein expression on the cell surface of CTC. A variety of microfluidics and filtration devices has been developed and are currently under evaluation for selection and enumeration of CTCs. CTC detection and molecular characterization systems are mainly based on protein and image-based approaches like classical immunocytochemistry, the FDA cleared CellSearch system, and immunofluorescence, and molecular assays based on the nucleic acid analysis in CTCs like RT-qPCR, multiplex RT-qPCR, and next generation sequencing technologies. Quality control and standardization of CTC isolation, detection and molecular characterization methodologies is very important for the incorporation of CTCs into prospective clinical trials testing their clinical utility. CTC molecular characterization at the single cell level holds considerable promise for the identification of therapeutic targets and resistance mechanisms in CTCs as well as for the stratification of patients and real-time monitoring of systemic therapies. This lecture will be mainly focused on the analytical systems for CTC isolation, enumeration, and detection and the clinical applications of CTC in many types of solid cancer. We also discuss the potential of the molecular characterization of CTC as a liquid biopsy in individualized therapy.






Biography:

Abstract:
Background: Comet assay is a quick method for assessing DNA damage in individual cells. It allows the detection of single and double DNA strand breaks which represent the direct effect of some damaging agent. This work aims to use comet standard quantification models to compare the neurotoxic effect of orally administered propionic acid (PA) to that produced as metabolite of bacterial overgrowth induced by clindamycin. Additionally, the protective effect of carnosine and carnitine as two natural dietary supplements was assessed. Methods: Single cell gel electrophoresis (comet assays) were performed on brain cortex and medulla after being removed from the nine studied groups of hamsters, served as control, PAintoxicated, clindamycin- treated together with carnosine and carnitine treated or protected groups. Results: The obtained data show a significant double strand breaks recorded as tail length; tail moment and % DNA damage in PA and clindamycin-treated brain cortex and medulla compared to control-untreated hamsters. Moreover it proves the neuroprotective effect of carnosine and carnitine. Receiver Operating Characteristics curve (ROC) analysis show satisfactory values of sensitivity and specificity of the comet assay parameters. Conclusion: Percentage DNA damage, tail length, and tail moment are feasible to be used as biomarkers of PA neurotoxicity when given either orally or as metabolite of induced enteric bacterial overgrowth. Establishing biomarkers for these two exposures is of importance to protect the health of children and could be helpful in controlling the prevalence of autism, a disorder recently related to PA neurotoxicity.






Biography:

Abstract:
Altered mitochondrial function and free radical-mediated tissue damage have been suggested as important pathological events in isoproterenol (ISO)-induced cardiotoxicity. This study was undertaken to know the preventive effect of moin on mitochondrial damage in ISO-induced cardiotoxicity in male Wistar rats. Myocardial infarction in rats was induced by isoproterenol (85 mg/kg) at an interval of 24h for 2 days. Morin was given to rats as pretreatment for 30 days orally using an intragastric tube. Isoproterenol caused a significant increase in the activity of cardiac injury marker enzymes like CK and LDH. Morin pretreatment significantly reduced the concentration of CK and LDH. ISO-treated rats had showed a significant elevation of mitochondrial TBARS and HP level and Pre-treatment with morin significantly prevented the increase of TBARS and HP level to near normality. The level of enzymic and non-enzymic antioxidants was decreased significantly in ISO-treated rats and Pre-treatment with morin significantly increased the levels of SOD, CAT, GPX, GST and GSH to normality. The activities of mitochondrial enzymes such as isocitrate dehydrogenase (ICDH), -ketoglutarate dehydrogenase (-KGDH), succinate dehydrogenase (SDH) and malate dehydrogenase (MDH) were decreased significantly in ISO treated myocardial ischemic rats and upon pre-treatment with morin restored these enzymes activity to normality. In addition, the decreased activities of cytochrome c oxidase and NADH-dehydrogenases were observed in ISO-treated rats and pre-treatment with morin prevented the activities of cytochrome c oxidase and NADH-dehydrogenase to normality. Pre-treatment with Morin favorably restored the biochemical and functional parameters to near normal indicating morin to be a significant protective effect on cardiac mitochondrial function against ISO induced myocardial infarction in rats.






Biography:

Abstract:
The aim of this investigation was to evaluate the preventive role of morin, a flavonoid, on cardiac marker enzymes such as aspartate transaminase, lactate dehydrogenase, creatine kinase and creatine kinase-MB, membranebound enzymes such as sodium potassium-dependent adenosine triphosphatase, calcium-dependent adenosine triphosphatase and magnesium-dependent adenosine triphosphatase, and glycoproteins such as hexose, hexosamine, fucose and sialic acid in isoproterenol (ISO)-induced myocardial infarction (MI) in rats. Male albino Wistar rats were pretreated with morin (20, 40 and 80 mg/kg) daily for a period of 30 days. After the treatment period, ISO (85 mg/kg) was subcutaneously injected into the rats at an interval of 24 h for 2 days. ISO-induced rats showed significantly (P.05) increased activities of cardiac marker enzymes in serum and decreased activities in the heart, and increased activities of calcium-dependent adenosine triphosphatase and magnesium-dependent adenosine triphosphatase in the heart, and the activity of sodium potassium-dependent adenosine triphosphatase decreased in the heart. ISO induction also showed a significant increase in the levels of glycoproteins in serum and the heart. Pretreatment with morin (40 mg/kg) daily for a period of 30 days exhibited significant (P.05) effects and altered these biochemical parameters positively compared to the other two doses. Thus, our study shows that morin has a protective role in ISO-induced MI in rats. The observed effects might be due to the free radical-scavenging, antioxidant and membrane stabilising properties of morin.






Biography:

Abstract:
The present study was designed to investigate the preventive effect of morin on lysosomal enzymes in isoproterenol (ISO) treated myocardial infarcted rats. Myocardial ischemia was induced by subcutaneous injection of ISO hydrochloride (85 mg/kg BW, twice at an interval of 24h) for two consecutive days. The morin (40 mg/kg BW) was administered daily for 30 days and subsequently two doses of ISO administered on 29th and 30th days. The activities of lysosomal enzymes β-glucuronidase, β-N-acetylglucosaminidase, β-galactosidase, cathepsin-B and D were increased significantly (P<0.05) in the serum and heart of ISO-induced cardiotoxic rats. The pretreatment of morin and two doses ISO treated rats exhibited significant (P<0.05) reduction of these lysosomal enzymes activities. Morin protects the lysosomal membrane damage against ISO-induced cardiac damage as evidenced by reduced activities of these lysosomal enzymes.






Biography:

Abstract:
Carvacrol (2-methyl-5-(1-methylethyl)-phenol) is a predominant monoterpenic phenol which occurs in many essential oils of the family Labiatae including Origanum, Satureja, Thymbra, Thymus, and Corydothymus species. It is well known for its anti-inflammatory, antioxidant and antitumor activities. The present study investigates the influence of carvacrol on CYP2E1 and PPAR-α on D-GalN-induced hepatotoxic rats. The mRNA and protein expression levels of CYP2E1 and PPAR-alpha have been assayed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot analysis. We found that the mRNA and protein expressions of CYP2E1 significantly up-regulated while the mRNA and protein expressions of PPAR-alpha significantly down-regulated on D-galactosamine induced hepatotoxic rats and treatment with carvacrol significantly suppressed the mRNA and protein expressions of these genes. Thus, the present results have shown that carvacrol has the hepatoprotective effect and also alleviates liver damage associated with GalN induced hepatotoxic rats by down-regulating the CYP2E1 and up-regulating the PPAR-alpha expression.






Biography:
Hala M. Abdelkarem has a special interest in obese, diabetes, osteoporosis and nutritionally-related health problems. Able to explain their pathophysiology. Special interested in aspects of numerous biochemical analysis in biological fluids. Teach lecture of biochemistry, estimation of nutritional elements, vitamins and hormones in blood fluids. The chemical and biological evaluation in dietary nutrition. Studies of different methodology to evaluate the nutrition status in patients. Estimation of bone markers in the biological fluids. She is the member of nutrition council at minster of Health, Egypt. Member of scientists council, Egypt. Member molecular biology and biochemistry council, Faculty of medicine, Ain Shams University, Egypt. Member of diabetic and endocrinology council, Diabetic institute, Minister of Health, Egypt. Member of medicinal council, National Research Center, Egypt.
Abstract:
Background: The metabolic syndrome (MetS) is a constellation of risk factors, including impaired fasting glucose, hypertension, central adiposity, predisposing to higher risks of oxidative stress, type 2 diabetes and atherosclerotic cardiovascular disease (CVD). Obesity/insulin resistance is associated with metabolic syndrome, which plays a pivotal role in cardiovascular risk. The mechanisms that link obesity, insulin resistance, and endothelial dysfunction are numerous and complex Increase in visceral fat, usually involved in obesity, leads to an imbalanced production of metabolic products, hormones, and adipocytokines including tumor necrosis factor-alpha (TNF-alpha), free fatty acids (FFAs) or adiponectin which causes decreased insulin sensitivity in skeletal muscle and liver, and impairs endothelial function through direct or indirect mechanisms.
Objective: The purpose of this study is to assess the beneficial effect of quercetin, flaxseed and/or in combination as synergetic in an animal model of metabolic syndrome (MtS), high fructose (HF) -fed rats.
Methods: Fifty male Sprague-Dawley rats, 3-month old and weighing between 110-120g were randomly divided into 5 groups. Rats were given drinking water (-ve control rats) or 10% fructose in drinking water (HF; fructose-fed rats) with standard chow for 8 wk. After 4-wk HF feeding, rats were further divided into matched 4 subgroups. Different groups of animals (n-10, each group) were administered; 10% HF (5 mg/kg, + ve control), flaxseed (F; 50 mg/kg), quercetin (Q; 50 mg/kg), flaxseed+quercetin, (FQ; 25 mg/kg of each) respectively. All ingredients were given orally once daily and subsequent 4 wk. Serum glucose, insulin, lipids profile, leptin, and adiponectin were estimated.
Results: After 4 weeks of feeding, a significant increase in blood glucose level was observed in HF fed rats compared to normal rats, but this increase was significantly decreased after administration of F, Q & FQ. The raised of serum insulin level in HF fed rats was significantly decreased after administration of F and FQ groups. Significantly higher concentrations of triacylglycerols (TG), total cholesterol & low density lipoprotein cholesterol (LDL-C) were observed in HF fed rats and these increases were lower after administration of F, Q & FQ. There was a significant increase in serum high density lipoprotein cholesterol (HDL-C) in FQ group. The increased of serum leptin level was decreased significantly in F, Q & FQ groups. Whereas the reduction of serum adiponectin level in HF fed rats was increased in F, Q & FQ groups.
Conclusion: These data suggested that protective effect of flaxseed and quercetin consumption as functional foods could be reduced risk for people with decreased insulin sensitivity and increased oxidative stress, such as those with the metabolic syndrome or type 2 diabetes.