Biography:

Maria Longas completed her PhD at New York University in 1978, and did her postdoctoral training at Columbia University School of Medicine with Dr. Karl Mayer. She has an MA in Organic Chemistry from New York University (1973) and a BA in Chemistry from Hunter College (1971). She is a full Professor of Chemistry at Purdue University Calumet in Hammond, IN. She has more than 24 papers published in reputable journals, and served as a reviewer for several journals.

Abstract:

Ultraviolet light B (UVB) is a technique used to investigate organic molecules. It was utilized in this study to analyze the content of vitamin D3 in face lifted female skin of subjects aged 43, 47, 51, 58, 60 and 66 years. The skin of these subjects had pigmentation of great similarity. It was kept at -40O C, defrosted to room temperature for 2 min and placed in an oven for another 2 min at 105o C. After these 2 min, we took the first UVB reading; a 2nd UVB run was done at room temperature. The amount of vitamin D3 was determined under standard conditions of 7-dehydrocholesterol (7-Dchol). The best temperature to measure D3 was when skin was at room temperature for 15 min. The D3 analysis of the UVB plots of human skin vs. the standard showed a linear decrease in human skin as compared to 7-Dchol. Because 7-Dchol is a direct precursor of vitamin D3, it appears from these results that as the skin ages, 7-Dchol concentration decreases which results in less production of vitamin D3. Conditions to increase vitamin D3 in the aged should be considered, to avoid problems of arthritis, tuberculosis and autoimmune disorders, etc.

Biography:

Dr. Razi is the founder of AccuDava Inc., a biomedical company for personalized cancer chemotherapy (http://www.accudava.com), based on the discovery of two Glycomarkers.  Razi received her PhD in Medical Sciences from Uppsala University-Sweden, in 1995. In her post-doctoral program, at the University of California-San Diego, she discovered a novel glycan-mediated lymphocytes activation mechanism in the immune system. She then joined the Consortium for Functional Glycomics (CFG), at The-Scripps-Research-Institute, where her team successfully developed and launched the world’s largest mammalian glycan microarray in 2011. This unique platform that displays 611 defined glycans has since become available globally for studying glycan-mediated interactions. 

Abstract:

Personalized chemotherapy is an unmet need in cancer treatments. A molecular test that can accurately predict the drug efficacy on a patient would be invaluable in selecting effective treatment strategy.

            We have identified two glycan structures, namely Glycomarker-1 and Glycomarker-2, whose expression levels on the cancer cell surface are associated with responses to chemotherapy with platinum drugs (US patent #7585503, and International pending patent). This finding introduces a new concept, linking glycan cell surface expressions with drug reactivity, and proposes a glycan-mediated mechanism for drug uptake.

            Our initial studies profiling the cell surface glycans, using Flow Cytometry with specific glycan-binding lectins, on three isogenic pairs of ovarian carcinoma cell-lines, consisting of chemosensitive and acquired chemoresistant phenotypes, revealed at least a ten-fold decrease in alpha2-6Sialyl-R motif (Glycomarker-1) on resistant phenotypes compared to sensitive cells. Further studies by fluorescent confocal microscopy, colony forming assay, sialidase treatments, and mass-spectrometry confirmed the association of Glycomarker-1 with drug uptake.

            Using Lectin histochemistry (LHC) on clinical samples proved a feasible assay for Glycomarker-1, tested on 64 human ovarian normal and cancerous tissue sections. The LHC on retrospective ovarian cancer specimens, with a known history of drug-response, correctly predicted drug-responses in 22 out of 27 (81.4%) patients.

            During the studies on the mechanism for drug response, another glycan structure, Glycomarker 2, was identified demonstrating a similar expression pattern to Glycomarker 1. Further studies on Glycomarker-2 suggest an association of the two glycomarkers that would put forward a glycan-mediated mechanism for platinum-drug uptake by cancer cells.  

Biography:

Dr. Desai is the Director of Institute for Structural Biology, Drug Discovery and Development at Virginia Commonwealth University. He earned his PhD from Indian Institute of Technology, Bombay and did postdoctoral studies at Iowa, MIT and Illinois. He was awarded the Established Investigator Award by the American Heart Association and the Distinguished Scholarship Award by Virginia Commonwealth University.

Abstract:

Designing glycosaminoglycan (GAG) sequences that target specific proteins is challenging and not yet achieved. This is a key reason that nature’s library of millions of glycosaminoglycans (GAGs) sequences remains largely untapped. We have developed a very generalizable genetic algorithm-based dual-filter screening strategy that addresses this key hurdle. Our computational algorithm utilizes ‘affinity’ and ‘specificity’ filters and a combinatorial library of 46,656 heparan sulfate (HS) hexasaccharides binding to target protein(s) for identifying ‘highly specific’ HS sequences. Synthesis of these sequences followed by biochemical and biophysical studies help test the computational design predictions. This talk will focus on differential targeting of two highly similar proteins and will present design protocol for discovering high specificity GAG sequences. Considering that GAGs bind to several hundred human proteins, majority of which remain un-characterized with regard to GAG interactions, this work lays the pathway for discovering highly selective GAGs as drugs and/or chemical biology tools.

Biography:

Xueli Li completed her PhD from Leipzig University, Germany. She is a research associate in The Michael J Palmieri Metabolic Laboratory, Children’s Hospital of Philadelphia, Philadelphia, PA. She has more than 8 publications in the area of Glycomics recently.

Abstract:

Protein glycosylation is increasingly recognized as a crucial modulator of protein function, offering a third layer of biological information over genomics and proteomics. Modern tools for analyzing released N-glycans from cells, the glycome, have shown abnormal protein glycosylation in numerous human diseases. We developed a quantification of glycome in cells. Upon reaching 100% confluence, the cells were washed twice with PBS and harvested using a cell scraper. The cells were then pelleted and washed with PBS by centrifugation. Fibroblast pellets were lysed in more than 200 μl PBS, and 200 μg protein from the cell lysate was denature and precipitated with 2× volume of 100% propanol. N-linked and O-linked glycans were released from denatured protein, desalted and permethylated before subject to MALDI-TOF analysis. The quantification of O-linked glycans was achieved by spiking glycans from cell with 25μM of C13-labelled T antigen (m/z 543) and C13-labelled sialylated T antigen (m/z 909). N-linked glycans were quantified using 25μM of C13-labelled Man7GlcNAc2 (m/z 1107). Using this method, we identified abnormal fibroblast glycomes in a number of known patients with congenital disorders of glycosylation and demonstrates cellular glycome as an important tool for diagnosis of these diseases.

Biography:

Dr. Menter received his PhD degree in Chemistry from the George Washington University in 1969. He completed a postdoctoral fellowship with Prof. Dr. Theodor Foerster at the Institut fuer physikalische Chemie der Universtiaet Stuttgart, Germany. Subsequently, he was at the University of Alabama, Birmingham, and the VA Medical Center (Atlanta) He currently serves as Research Professor of Biochemistry at Morehouse School of Medicine. Dr. Menter is recognized internationally for his work in the areas of collagen photochemistry and melanin photobiology as pertaining to redox reactivity

Abstract:

Mammalian collagens exhibit weak intrinsic age- and history-dependent UV fluorescence that. UV radiation can cause longer wavelength fluorescent oxidative bands. The fluorescence of both unirradiated and UVC – irradiated acid-soluble calf skin collagen also reflects considerable autoxidation. We have been studying the ground-and excited state behavior of type I acid – soluble collagens from commercial calf skin and from acid-extracted hairless mouse skin as judged by their temperature and wavelength dependence of their fluorescence properties. As the age-related oxidation products destabilize the overall collagen supramolecular structure, we wondered whether the surrounding extracellular matrix affects the ground- and excited state behavior of the collagen polymer in situ. Added sodium hyaluronate (2:1 ratio) shows little or no effect on fluorescence behavior or Arrhenius – type plots. This latter result may indicate that there is little physical interaction between the collagen telopeptide and HA domains in vitro.

Biography:

Tongzhong Ju received his M.D. degree from the Qingdao Medical College
in1986 and his Ph.D. in Biochemistry from Fudan University Shanghai
Medical School (former Shanghai Medical University, China) in 1994. He
completed his post-doctoral training in Biochemistry/Glycobiology in 1999, worked as a Research Associate (1999-2002) and Research Assistant
Professor (2002-2006) at the University of Oklahoma Health Sciences
Center (OUHSC). Since 2006, Dr. Ju has been an Assistant Professor, and now an Associate Professor in the Department of Biochemistry at Emory University School of Medicine. He has published more than 50 papers in many different reputable journals.

Abstract:

Mucin–type O-glycosylation of Proteins is one of the most abundant protein post-translational modifications, and plays important roles in many biological processes, but the repertoire of mucin-type O-glycans synthesized by cells is difficult to determine. Here we developed a novel technology termed Cellular O-Glycome Reporter/Amplification (CORA), a sensitive and versatile method to profile and amplify mucin type O-glycans in any living cells. Cells took up peracetylated benzyl-α-N-acetylgalactosamine, Benzyl-α-GalNAc(Ac)3 into cytosol where it was de-acetylated to form Bn-α-GalNAc, and Benzyl-α-GalNAc was then transported into the Golgi apparatus where it was converted to a large variety of complex O-glycans (Benzyl-O-glycans) by O-glycosyltransferases; and these Benzyl-O-glycans were finally secreted from cells, allowing easy purification for analysis by HPLC and mass spectrometry (MS). CORA resulted in ~100-1000-fold increase in sensitivity over conventional O-glycan analyses, such as β-elimination-MS and identified a more complex repertoire of O-glycans in more than a dozen cell types analyzed. Thus, the high sensitivity and amplification nature of CORA technology makes it possible to analyze entire human O-glycome and others, and offers new opportunities to identify novel glycan biomarkers for human diseases. Furthermore, CORA technology may shed the light on facilitating the development of similar technologies for analyzing/amplifying other types of cellular glycomes.

Biography:

Jan Tkac received his Ph.D. degree in biotechnology at the Slovak University of Technology (Bratislava, Slovakia) in 2000. He received his postdoctoral appointment at the Linkoping University (2001-2003), Lund University (2003-2006) and Oxford University (2006-2008). Currently, he works at the Institute of Chemistry, Slovak Academy of Sciences in Bratislava. He was/is a recipient of an Individual Marie Curie Fellowship (2003) and an ERC Starting grant (2012). His main research activities cover a microarray platform of detection of glycoproteins by lectins with the aid of nanotechnology. He has published 80+ scientific articles, reviews, and 4 book chapters having 1000+ citations.

Abstract:

Integration of nanotechnology into the field of glycomics can address current limitations of fluorescent glycan and lectin microarrays [1,2]. Moreover, label-free transducing schemes especially when working in an ultrasensitive fashion can be applied in a reliable detection of low abundant disease biomarkers [3-5]. For example, electrochemical impedance spectroscopy applied in our studies allows to detect analyte molecules down to a single molecule level (i.e. aM level) if immobilization architecture of ligands (lectins or glycans) is controlled at nanoscale [6-8]. Utilization of such devices in serological glycoprofile of samples from people having some diseases, in analysis of cancer biomarkers, cancer cell lines and viruses will be discussed. Acknowledgement: The financial support from the Slovak research and development agency APVV 0282-11 is acknowledged. The research leading to these results has received funding from the European Research Council under the European Union’s Seventh Framework Program (FP/2007-2013)/ERC Grant Agreement No. 311532 and from the European Union’s Seventh Framework Program for research, technological development and demonstration under grant agreement No. 317420.

Biography:

Dr. Ming Zhang received the Ph.D. degree (Summa Cum Laude) from University of Gottingen, Germany. She has over a decade experience in infectious diseases, in particular targeting vaccine designs for HIV and influenza viruses. She is an Assistant Professor of Epidemiology at University of Georgia, USA, and leads an interdisciplinary research group towards understanding pathogen diversity and mechanisms for vaccine design purposes. She is also keen in educating the next generation of critical thinkers and innovators in infectious diseases.

Abstract:

N-linked glycosylation is a posttranslational modification that has significantly contributed to the rapid evolution of HIV-1. In particular, enrichment of N-linked glycosylation sites can be found within Envelope variable loops, regions that play an essential role in HIV pathogenesis and immunogenicity. In this presentation, I will describe some strategies we learned through though studies of HIV N-linked glycosylation, on both individual and population aspects, that can be implemented in virus vaccine designs, including for influenza prevention purposes.

Biography:

Robert Sabatini has completed his PhD at the age of 30 years from the University of Alabama at Birmingham and postdoctoral studies from the Netherlands Cancer Institute (Amsterdam). He is an Associate Professor at the University of Georgia in the Department of Biochemistry and Molecular Biology. He has published more than 30 papers in reputed journals.

Abstract:

Kinetoplastids are a group of early-diverged eukaryotes that includes the human parasites Trypanosoma brucei, and Leishmania major. The genome of these pathogens contains a modified DNA base (beta-D-glucosyl-hydroxymethyluracil) consisting of O-linked glucose modification of the thymine base such that the glucose moiety sticks out into the major groove of DNA. The modified base, called base J, is synthesized through the hydroxylation of thymidine by a dioxygenase (JBP) forming hydroxymethyluridine, followed by the transfer of a glucose by the glucosyltransferase enzyme JGT. I will present our most recent studies on the synthesis and function of base J, focusing on its role in regulating RNA polymerase II (RNAP II) transcription termination. The genomes of kinetoplastids are organized into polycistronic gene clusters that contain multiple genes that are co-transcribed from a single promoter. We have localized base J at regions flanking the polycistronic clusters at sites involved in initiation and termination and demonstrated that the loss of J results in increased RNAP II recruitment at promoters and transcription of the clusters. We now show that reduction of base J at termination sites within polycistronic gene clusters leads to readthrough transcription and increased expression of downstream genes, including developmentally regulated genes involved in parasite pathogenesis. The current model is that the glucose moiety of base J provides a steric block for transcription elongation stimulating termination. The role of the GTase JGT in regulating the localization of J at specific sites in the genome will also be discussed.

Biography:

Ying Xu is the "Regents and Georgia Research Alliance Eminent Scholar" Chair Professor in Biochemistry and Molecular Biology Department and the Founding Director of the Institute of Bioinformatics, the University of Georgia since 2003. Before that, he was a senior staff scientist and group leader at Oak Ridge National Laboratory. He received his Ph.D. degree in computer science from the University of Colorado at Boulder in 1991. His current research interests are in cancer systems biology and microbial genomics. He has over 300 papers and five books. He is currently the Editor-in-Chief of IEEE Transaction on Computational Biology and Bioinformatics.

Abstract:

We have recently conducted a computational analysis of transcriptomic data of all enzymes involved in the synthesis of glycans of 20 types of cancer versus matching control tissues. Our analyses revealed a number of highly interesting results, including that these glycans predominantly are involved in balancing hydroxyl radicals produced by the diseased cells and some of their oxidized products drive a number of activities typically associated with cancer initiation and development.

Biography:

KLAUS FIEDLER has completed his PhD at the age of 27 years following a Master of Science with a degree in Biology II (Biochemistry) from EMBL Heidelberg with Prof. K. Simons and the Basel University. He carried through his postdoctoral studies at the Rockefeller Research Laboratories, Sloan-Kettering Institute with Prof. J. Rothman. He held a START research grant from the Swiss National Science Foundation thereafter. His first research projects in biological sciences were followed by medical studies at the University of Basel which led to a Master of Medicine. He has published more than 25 papers in reputed journals, monographs and book chapters.

Abstract:

The modeled Golgi dynamics domain of CHOp24, a member of intra-Golgi cargo receptors, scores highly with a collagen-binding domain and carbohydrate-binding modules and is shown to interact with Wnt8 (wingless 8) with a G=-18.3 kcal/mol. Hybrid N-glycans may provide increasing energy of binding up to -7.1 kcal/mol to simulated p24-Golgi dynamics domain-ligand interaction. Apical transport may require N- and O-glycans and thus the interaction may offer insight on epithelial polarity. Groups of regulated molecules in epithelial polarization may include the sorting machinery of epithelial cells, sorted ligands or both, on top of the intricate regulatory mechanisms in substrate and intercellular adhesion. In further research I have analyzed the cargo receptor VIP36 (Vesicular-integral membrane protein of 36 kDa) for carbohydrate interaction. It has been described by me as a lectin in the endoplasmic reticulum-Golgi intermediate compartment, Golgi apparatus and plasma membrane and later was found implicated in parotid gland secretion and apical transport in MDCK cells. The docking reveals top-interacting carbohydrates of the N-glycan and O-glycan class that encompass N-linked glycans of proteins of high mannose and equally complex type which likely function as sorted ligands in epithelial cells. High affinity binding of the ganglioside GM1 carbohydrate headgroup to VIP36 suggests a linkage with protein and glycosphingolipid apical transfer in epithelial cells. Thus, this fundamental approach predicts that interchangeable glycosphingolipid/protein cargo receptor interaction, which may include some p24 family members in glycan binding, may function in apical transport.