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Here are the outstanding speakers you will hear at Planet xMAP USA 2010:

Keynote Speakers

Keynote Address - Wednesday, May 12

Dr. James D. Watson, Ph.D.
Nobel Prize-Winning Scientist and Co-Discoverer of the DNA Structure

Presentation:  Rules for Science

James D. Watson Bio:
James D. Watson was born in Chicago, Illinois in 1928 and educated at the University of Chicago.  In 1953, while at Cambridge University, he and Francis Crick successfully proposed the double helical structure for DNA, a feat described by Sir Peter Medawar as "the greatest achievement of science in the twentieth century."  For this work, Watson and Crick, together with Maurice Wilkins, were awarded the Nobel Prize in Physiology or Medicine in 1962.  While a Professor at Harvard, Watson commenced a writing career that generated the seminal text, Molecular Biology of the Gene, the best-selling autobiographical volume, The Double Helix, and most recently Avoid Boring People.  Later, while leading the Cold Spring Harbor Laboratory, he was a driving force behind setting up the Human Genome Project, a major factor in his receipt in 1993 of the Copley Medal from the Royal Society that elected him a member in 1981.  Among other honors, Watson was elected in 1962 to the National Academy of Sciences and, in 1977, received from President Ford the Medal of Freedom.  He has received honorary degrees from many universities including The University of Chicago (1961), Harvard University (1978), Cambridge University (1993), University of Oxford (1995), Trinity College, Dublin (2001), and Uppsala University (2007).  Dr. Watson received the National Medal of Science in December 1997, the City of Philadelphia Liberty Medal on July 4, 2000, and the Benjamin Franklin Medal awarded by the American Philosophical Society in 2001.  Queen Elizabeth II proclaimed him an honorary Knight of the British Empire on January 1, 2002.  Dr. Watson has served the Cold Spring Harbor Laboratory in several capacities since 1968 [Director 1968-1994, President 1994-2003, Chancellor 2003-2007] and now is Chancellor Emeritus.

Keynote Address - Thursday, May 13

Linette Granen, MT (ASCP), DLM
Manager, Corporate Relations, Association of Public Health Laboratories

Presentation: Public Health Laboratories and Luminex:  Anatomy of a Partnership

Abstract:
At the end of this presentation, the participants will be able to

1.  Outline the role of the public health laboratory community in the US healthcare system;

2.  Describe the public-private partnership that exists between APHL and Luminex     Corporation; and

3.  List the uses of the Luminex technologies in public health laboratories.

The Association of Public Health Laboratories (APHL) uniquely represents state and local public health, environmental, agricultural and other public sector laboratories.  The association serves as a liaison between member laboratories, federal government agencies such as CDC and FDA, and private sector partners, like Luminex Corporation.  Public health laboratories are on the front line in monitoring and detecting public health threats.  In addition, they screen newborns for genetic and metabolic disorders, monitor and respond to threats in the environment, conduct testing supporting water, food, dairy and environmental safety laws, research disease trends, develop new laboratory technologies and contribute to the formulation of state and national health policies.  APHL has a successful corporate membership program that is becoming a model of public-private partnership for the non-profit community, where industry interacts with the membership of the association toward achieving our vision of worldwide quality laboratory practice. The unique mandate of public health laboratories that blends clinical laboratory detection technology and surveillance/research laboratory applications with governmental guidelines presents a challenge for our private partners.  Luminex has risen to that challenge, supporting public health laboratories in supplying technology for infectious disease testing, food safety, newborn screening and biodefense analyses.  In addition, through the increased level of communication that exists between the company and the association, support for APHL’s workforce initiatives and continuing education conferences has been provided.  At least one case study will be cited as an example of how Luminex technology has contributed to the “Analysis” and “Answers” segments of our slogan, “Public Health Laboratories—Analysis, Answers, Action”.

Linette Granen Bio:
Linette Granen, MT(ASCP), DLM  is the Corporate Relations Manager for the Association of Public Health Laboratories (APHL), which represents the state, local and county public health, environmental health and agricultural laboratories that protect the health and safety of the public.  She is responsible for partnering with industry through the APHL Sustaining Member Program, which has been very successful since its inception in 2005.  Having worked at APHL for the past 16 years, Ms Granen also held the titles of Marketing Manager and Regional Manager for the National Laboratory Training Network, a joint program of APHL and CDC.  Prior to working for APHL, Ms. Granen was the Program Manager for Diagnostic Laboratory Services and Herd Health for Mississippi State University College of Veterinary Medicine.  Throughout the rest of her 35 year career, as a licensed medical technologist and diplomate in laboratory management, she has served in laboratory technical and management roles in small hospitals, large medical centers and physician office laboratories. 

Keynote Address - Friday, May 14

Dr. Todd R. Golub
Director, Cancer Program, BROAD Institute of Harvard and MIT

Presentation: Bead-based Approaches to Cancer Profiling 

Abstract:
Unbiased surveys of the cancer genome and cancer proteome can provide powerful new insights into the pathogenesis of cancer.  Toward that end, we have developed a number of molecular profiling approaches that utilize Luminex beads as a high-throughput, low-cost approach.  In particular, we developed a highly multiplexed phosphotyrosine profiling assay capable of profiling the phosphorylation status of most human tyrosine kinases.  We used this method to discover the aberrant activation of SRC kinase in glioblastoma, among other kinase activation events.  In addition, we have developed methods for the pooling of cancer cell lines in a single well of a multi-well plate, with deconvolution of the cell line identities using molecular barcodes detected on Luminex beads.  Lastly, we have developed highly multiplexed mRNA assays to monitor gene expression, with assays containing up to 1000 transcripts per well, and we demonstrate that this approach can be used to screen for small-molecule modulators of biological processes of interest.    

Todd R. Golub Bio:
In 1997, Todd joined the Whitehead Institute/MIT Center of Genome Research, now part of the Broad Institute, as director of its program in cancer research. He is the Charles A. Dana Investigator in Human Cancer Genetics at the Dana-Farber Cancer Institute, an investigator at Howard Hughes Medical Institute, and professor of pediatrics at Harvard Medical School. Todd is the recipient of multiple awards, including the 2009 AACR Richard and Hinda Rosenthal Memorial Award, the 2008 E. Mead Johnson Award from the Society for Pediatric Research, the 2007 Oski Prize of the American Society of Pediatric Hematology-Oncology, Discover Magazine's Inventor of the Year (Health Category), the Daland Prize of the American Philosophical Society, the Outstanding Achievement Award for the American Association for Cancer Research, and the 2007 Paul Marks Prize for Cancer Research. Todd also serves on the Board of Scientific Advisors of the National Cancer Institute.

Todd received his B.A. in 1985 from Carleton College and his M.D. in 1989 from the University of Chicago Pritzker School of Medicine.  

Sally Lewis, Ph.D.
Department Head Clinical Laboratory Sciences
Tarleton University

Presentation: Adventures in Genotyping:  Incorporation of Luminex  Cytochrome Genotyping Assays into a Medical Laboratory Science Curriculum

Abstract:
Our Medical Laboratory Sciences (MLS) program has incorporated molecular diagnostics/molecular biology theory into the curriculum for over a decade. In 2007, with the establishment of a state-of-the-art molecular diagnostics laboratory, and the addition of a talented molecular scientist (Dr. Heping Han) to our faculty, our department was able to provide molecular laboratory exercises for our students.  Incorporation of real-time PCR exercises with validation testing to our student laboratories, was important in exposing students to the high complexity laboratory testing that will play a prominent role in laboratory diagnostics.  A research project involving cytochrome p450 genotyping has allowed us to expose our students to Luminex genotyping technology and the establishment of a new technology into our laboratories.  Graduates of our MLS program have already benefited from this exposure by securing positions in molecular research and clinical molecular diagnostics laboratories.  We hope that future graduates will benefit from their genotyping experiences with employment by industry partners.

Sally Lewis Bio:
Sally Lewis received her B.S. in Zoology and Physiology from Louisiana State University, her M.S. in Biology with an emphasis in Immunology from Texas Christian University   and her Ph.D. in Molecular Biology from the University of North Texas .  She currently serves as Department Head and Program Director of the Medical Laboratory Sciences (MLS) Department at Tarleton State University in Fort Worth, Texas. Dr. Lewis has numerous years of clinical laboratory experience as Supervisor of Histology, Cytology and the clinical laboratory, and holds current certifications by the American Society of Clinical Pathology in Medical Laboratory Science (MLS), Molecular Biology (MB), and Histotechnology (HTL). She is an active member of several professional organizations including the American Society of Clinical Pathology, the American Society for Clinical Laboratory Science (ASCLS), the Texas Society for Clinical Laboratory Science (TACLS) and the American Society for Molecular Pathology (AMP). Dr. Lewis also serves as a Research Co-investigator in Medical Services at the Veterans Administration North Texas (VANT) Health Care System and as Adjunct Assistant Professor at the University of Texas at Austin. Dr. Lewis coordinates development activities for the May Owen Foundation, a private foundation whose sole mission is the support of the MLS department, and serves as a pharmacogenetic advisor to the VANT Cardiovascular Research Institute. 

John SantaLucia Jr., Ph.D.
Professor of Chemistry, Wayne State
CSO DNA Software, Inc.

Presentation:  Physical Principles and Software for Optimal Multiplex PCR Design

Abstract:
Optimal design of PCR reactions is a challenging problem, particularly as more complex formats such Multiplex PCR are introduced.  Secondary structural folding of DNA or RNA targets can inhibit hybridization causing false-negative PCR.  The presence of competing genomic DNA or contaminating organism genomic background can cause false-positive PCR.  Optimal Multiplex PCR design requires all probes/primers to be optimally sensitive and selective for their intended targets at the same reaction conditions.  Moreover, Multiplex PCR design must avoid destructive competing events such as target mis-hybridization, oligonucleotide cross-hybridization, and production of false amplicons.  This seminar will present some fundamental physical principles of DNA hybridization and folding and show how these principles apply to designing optimal Multiplex PCR reactions.  In addition, this seminar will highlight DNA Software’s novel Visual OMP™, ThermoBLAST™, and Modifieds™ software programs, which significantly improve Multiplex PCR performance.

DNA Software’s Visual OMP™, ThermoBLAST™, and Modifieds™ technologies contain advanced science and capabilities that are ideally suited for Multiplex PCR design.  Visual OMP™ incorporates its own dynamic programming algorithm for folding and secondary structure analysis.  In addition, Visual OMP™ uniquely contains an advanced algorithm for multi-state coupled equilibrium.  Accordingly, it folds and accounts for every possible interaction for each sequence in an experiment using actual solution conditions.  Moreover, Visual OMP™ produces cross-hybridization tables and graphs of Tm versus concentration for every possible sequence interaction.  Visual OMP™ also uniquely incorporates PCR additives (e.g. Luminex buffer, TMAC, and Betaine) plus many fluorophores and quenchers.  DNA Software uniquely offers ThermoBLAST™ and Modifieds™, which are two additional programs that significantly improve assay specificity and sensitivity.  ThermoBLAST™ was developed under a contract from the Department of Homeland Security for biodefense applications.  It combines the database capabilities of BLAST with the accurate thermodynamics of Visual OMP.  ThermoBLAST™ quickly and accurately scans DNA and RNA probes against large genome databases and discovers thermodynamically stable hybridizations and thereby eliminates assay false positives and off-target effects.  Modifieds™ designs assays, diagnostics, and therapeutics that incorporate modified nucleotides (e.g. LNA, PNA, Morpholino, DeoxyU, 5-methyl-C, Inosine, plus many more). 

John SantaLucia Jr. Bio:
Dr. John SantaLucia, Jr. is the co-founder and Chief Scientific Officer of DNA Software, Inc.  Dr. SantaLucia is also Professor of Chemistry at Wayne State University.  He is a leading scientist in the field of nucleic acid thermodynamics and RNA structural biology.  Dr. SantaLucia’s research has been cited by over 4000 publications.  He has served on numerous NIH study section panels and scientific advisory boards for several companies.  His academic research has focused on (1) thermodynamic analysis of DNA and RNA, (2) 3D structure prediction and analysis of RNA and DNA, (3) NMR structure determination of RNA with bound drugs and proteins, and (4) nucleic acid-based nanotechnology.  Dr. SantaLucia’s research is the foundation of many of DNA Software’s technologies, which have become the standard of excellence for nucleic acid-based research and diagnostics development. 

DNA Software, Inc. uniquely combines original wet lab research with pioneering software programs.  The company’s software tools are exceptionally accurate and optimally design and simulate complicated experiments such as Multiplex PCR on the first attempt.  DNA Software’s tools have helped hundreds of customers in diverse industries around the world to quickly develop optimally sensitive/selective nucleic acid-based assays, diagnostics, and therapeutics.  Luminex, the CDC, the FDA, and many other prominent research organizations utilize DNA Software’s tools for their Multiplex PCR design.  DNA Software, Inc. licenses off-the-shelf software for secondary structure analysis, assay development, genome scanning, and modified nucleotides.  DNA Software, Inc. also offers contract research, custom software and commercial web applications development, and scientific consulting for nucleic acid-based research.  DNA Software has recently expanded its R&D to include oligonucleotide kinetics and structural biology.  For more information, please contact DNA Software at +1 734 222 9080, info@dnasoftware.com or www.dnasoftware.com.

Millipore End-user Presentation

Jeffrey A. Borgia, Ph.D.
Assistant Professor
Rush Biomarkers Core

Presentation:  A Systems Biology Approach to Treatment Selection for
Non Small-Cell Lung Cancer Patients

Abstract:
Lung cancer is the #1 cause of cancer-related deaths in the worldwide, accounting for more deaths annually than cancers of the breast, prostate, colon and pancreas combined. Non small-cell lung cancer comprises the largest histological subtype of this disease and was responsible for the deaths of more than 160,000 Americans in 2009.  Improved molecular diagnostic assays are desperately needed to help reduce mortality of this dreaded disease and improve survival. Currently, there are very limited molecular insights that can assist clinicians facing critical treatment decisions for this disease. The three most decisive points impacting survival for these are (in order of disease progression): disease detection for asymptomatic patients, disease metastasis to locoregional lymph nodes, and optimal chemotherapeutic selection based on molecular phenotyping. Using a systems biology approach, we are developing a range of molecular diagnostics that will ultimately help stratify patients with NSCLC at these key decision points. In my lecture several panels of serum biomarkers will be presented that represent the beginnings of a system to better classify early-stage patients into distinct treatment groups which are aimed at pairing molecular characteristics of the patient’s tumor with clinical modalities offering the best prospects for a cure. Further, I will also present data that will permit the stratification of patients with advanced-stage disease to indicate the specific phenotypic characteristics of their tumor and which targeted molecular therapy stands to provide the most benefit. Included in this discussion will be the global assessment of tyrosine signaling cascades using Luminex-based immunoassays.


Jeffrey A. Borgia Bio:
Dr. Borgia is currently an Assistant Professor in the Departments of Biochemistry and Pathology at Rush University Medical Center in Chicago, Illinois. He is also the director of the Rush Proteomics and Biomarkers Core Facility where he assists investigators from around the world perform discovery and quantitative proteomic experiments as well as measure biomarkers using the Luminex platform for clinical trials. He is certified through the Luminex Corporation for Custom Immunobead Assay Development and Instrument Maintenance. Dr. Borgia is a member of the Rush Thoracic Oncology Group which is a research consortium made up of translational scientists, pathologists, pulmonologists, thoracic surgeons, and members from Radiation and Medical Oncology. His research examines molecular mechanisms underlying lung cancer metastasis and sensitivity/ resistance to systemic and targeted chemotherapies and is funded by the American Cancer Society, Illinois Chapter, LUNGevity Foundation, and the “Two-Years to the Cure” Foundation. Dr. Borgia originally trained with Dr. Theodore R. Oegema Jr. at the University of Minnesota in Minneapolis, MN and later was a Postdoctoral Scholar for Dr. Gregg B. Fields at Florida Atlantic University in Boca Raton, FL.  

Bio-Rad End-user Presentation

Jeremy Sokolove, M.D.
Postdoctoral Scholar
Division of Immunology and Rheumatology
Stanford University

Presentation: Multiplex Proteomics for Identification of Actionable Biomarkers in Rheumatoid Arthritis

Abstract:
Rheumatoid arthritis (RA) is the most common inflammatory joint disease of autoimmune etiology and remains a cause of substantial morbidity and mortality.  Current efforts in our laboratory focus on the development of tests for (i) the early diagnosis of RA, (ii) predicting the severity of RA, and (iii) guiding the selection of therapy for RA patients.  Our laboratory has developed and utilized RA antigen microarrays to profile the specificity of autoantibody profiles in RA.  We have now converted many of these biomarkers onto the clinical-grade BioPlex assay format, and are utilizing BioPlex technology for the high-throughput characterization of biomarkers in pre-disease, disease progression, and response to therapy cohorts.

Jeremy Sokolove Bio:
Jeremy Sokolove is a Clinical Instructor and Postdoctoral Scholar at Stanford University.  He attained an M.D. from Boston University where he was elected to the Alpha Omega Alpha Honor Society.  He then completed a residency at Boston University Medical Center where he was selected to serve as Chief Medical Resident.   Dr. Sokolove served as Brigade Medical Officer for the Marines 3^rd Force Service Support Group and as an attending Internist at Naval Medical Center San Diego where he was honored with a Naval Commendation Medal and received the Richard Daly Staff physician teaching award.  Dr. Sokolove subsequently completed a fellowship in Rheumatology at Stanford University where he continues as a post-doctoral scholar in the laboratory of Dr. William H. Robinson.  Dr. Sokolove is the recipient of the American College of Rheumatology Physician Scientist Development award.  His research interests are focused on biomarker characterization as well as mechanistic studies of autoimmunity in rheumatoid arthritis and related autoimmune conditions.

Jochen Schwenk, Ph.D.
Proteomics School of Biotechnology
KTH Royal Institute of Technology

Presentation:  Towards a Next Generation Plasma Proteome Profiling

Abstract:
To pursue an affinity-based proteome analysis of human body fluids, (i) high-throughput technologies such as microarrays and (ii) large collections of validated protein-specific capture molecules are required. Such challenging approach is now initiated at the Human Protein Atlas resource (www.proteinatlas.org), which has been set-up to allow a systematic, antibody-based exploration of the human proteome (1). With more than 10,000 well-characterized affinity reagents being accessible for a presented next generation profiling effort an antibody suspension bead array has been developed for biomarker discovery in serum and plasma.

A microtiter plate-based workflow has been developed that combines optimized buffer, sample labeling and treatment (2) to now perform up to 384 tests in parallel on 384 samples per day. This workflow does consume only minute amounts of serum or plasma (3), and allows for a multiplexed and sensitive protein profiling. The system has now already been utilized for various biomarker discovery projects, and antibodies have been selected in both directed and undirected fashion to reveal promising differences in protein profile between patient groups (data unpublished). The overall results suggested that the application of this emerging suspension bead array workflow holds the great potential to enable a proteome-wide, undirected biomarker discovery in a larger number of samples.

Therefore, we have recently started to profile human serum and plasma in a systematic manner: In a discovery phase antibodies are selected, validated and applied to screen a multi-disease cohort composed of 384 patient samples. Then, interesting findings are further to be qualified in subsequent phase with dedicated single disease cohorts. In the nearer future technological advances will allow to perform up to 1.000.000 assays per week and as more long-term objective develop streamlined assays based on two antibodies (dual specificity) to further increase the sensitivity and to evaluate the true potential of the biomarkers.

Jochen Schwenk Bio:
Dr. Jochen M. Schwenk is principal investigator for the plasma profiling group within the Human Protein Atlas project and working at the department of proteomics at the Royal Institute of Technology (KTH) in Stockholm, Sweden.

From the start of his scientific career, Dr. Schwenk was involved in protein microarray technologies developing a cell microarray for the analysis of cell surface specific antibodies during his master thesis. He continued with his doctoral thesis for Dr. Thomas Joos at the NMI in Germany, where he developed assays for bead-based and planar microarray applications with a focus on the characterization of affinity reagents and their application towards diagnostics. He received his PhD in Biochemistry in 2005 from the University of Tuebingen in Germany, from where he moved to the group of Prof. Mathias Uhlén’s at the Human Protein Atlas project. Here, he is involved in a broad range of protein array-based applications in the context of large-scale and multiplexed analysis of biobank material. Dr. Schwenk was recently nominated as “KTH star of 2009” for the WCN symposium on Frontiers in The New Biology.

Aravind Subramanian, Ph.D., MBA
Computational Biologist, Cancer Program
The Broad Institute of MIT & Harvard

Presentation: Analyzing the Entire Transcriptome with FLEXMAP 3D

Abstract:
Gene expression profiling can be used to identify molecular signatures that
describe cellular states of interest. Such signatures make it possible to describe
diverse biological states — from both experimental and clinical contexts — in a
common language. In this way, it is possible to systematically discover connections between small-molecules (drugs), genes (and their protein products) and diseases.
We will describe our efforts to generate ~100,000 gene-expression profiles of genetic
and pharmacologic perturbation based upon Luminex technology that is capable of simultaneously measuring the levels of up to 1,000 genes at modest cost. Combined
with novel computational algorithms, ‘Luminex-1000’ has the potential to reduce the
cost of gene-expression profiling by an order of magnitude, thereby empowering
the next generation of genomic applications.

Aravind Subramanian Bio:
Aravind Subramanian is a Computational Biologist in the Cancer Program at the Broad Institute. Aravind works with a diverse team of molecular biologists, research technicians, software engineers and analysts to develop new technologies and algorithms for mRNA profiling.

As a graduate student in the Whithead Institute/ MIT Center for Genome Research, Aravind helped develop Gene Set Enrichment Analysis (GSEA), a widely cited knowledge-based algorithm for the interpretation of high-dimensionality genomic datasets. In collaboration with colleagues in the RNAi platform at the Broad Institute, Aravind developed computational methods to analyze genome-scale pooled shRNA screens for the identification of essential genes in cancer cells (RIGER).

Aravind is currently working with members of the Todd Golub laboratory and Broad professional staff to implement a high-throughput, medium-density, low-cost gene expression-profiling platform based on Luminex beads.

Mark T. Esser, Ph.D.
Sr. Research Fellow, Operations
PPD Vaccines and Biologics Lab

Presentation: Multiplexed Assays for Multivalent Vaccines

Abstract:
Vaccine biomarker assays can be used  to monitor disease incidence and prevalence, measure immune responses to natural infection or vaccination and to determine the duration of immunity.  This talk will discuss how multiplexed assays are being used to support clinical trials and post-licensure surveillance studies of licensed and new multivalent vaccines.  Specific examples of how PPD Vaccines and Biologics Lab is employing molecular, antibody and cytokine assays on the Luminex platform to support vaccine studies for influenza, human papillomavirus and diphtheria, tetanus and pertussis vaccines will be presented.

Mark T. Esser Bio:
Mark Esser, Ph.D., is associate director of immunology at PPD’s vaccines and biologics lab, a leading facility for the comprehensive development and testing of vaccines and biologics. He brings to this position extensive experience in the development and validation of biomarker assays to support clinical development programs for vaccines and therapeutic proteins against infectious diseases and cancer.

Mark joined PPD in 2009 with the company’s acquisition of the Merck clinical vaccines and biologics laboratory. At Merck, he was part of the team that successfully achieved licensure of GARDASIL^® for the prevention of HPV infection. He represented Merck on several international HPV standardization committees and, in his current role at PPD, continues to actively collaborate with members from the World Health Organization, Centers for Disease Control and Prevention and the U.S. Food and Drug Administration on related issues.

Mark earned a bachelor’s degree. in biochemistry from Case Western Reserve University in 1992 and received his doctorate in microbiology and immunology from the University of Virginia. He served as a fellow at the National Cancer Institute in the AIDS vaccine program, where he was a member of a team that worked on the development of both a DNA vaccine and an HIV-1 virion-based vaccine for clinical evaluation.

Takeshi Shimamura, Ph.D.
Instructor in Medicine
Dana-Farber Cancer Institute

Presentation:  Epithelial to Mesenchymal Transition Phenotype in Non-small Cell Lung Cancer Cell Lines Contributes to EGFR Inhibitor Resistance

Abstract:
Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), gefitinib and erlotinib, lead to significant tumor regressions in approximately 10% of non-small cell lung cancer (NSCLC) patients with EGFR activating mutations. However, up to 40% of NSCLC patients, majority of whom are EGFR wild-type, do not benefit from EGFR TKI therapy. A mesenchymal phenotype is well-correlated with insensitivity to EGFR TKIs and often found in NSCLC cell lines with wild-type EGFR, though the molecular mechanisms still remain elusive. Here we find both tyrosine phosphorylation of ErbB family and Met receptors are downregulated in mesenchymal-like tumor cells. Additionally, mesenchymal-like NCI-H1975CLR cells, which was grown resistant to irreversible EGFR TKI, exhibit aberrant insulin receptor (IR) and IGF-IR expression and activation of the PI3-K and Src signaling pathways. Pharmacological inhibition of IR and IGF-IR receptors reduced cell proliferation and compromised cell survival in mesenchymal-like but not parental NSCLC cell lines, suggesting the switching of the predominant receptor tyrosine kinases controlling cell proliferation and survival. Using multiplex bead based quantification of 15 growth factors, we found deregulation of the expression of EGFR ligands and TGFβ1 in the mesenchymal-like cells. Mesenchymal-like transition in NCI-H1975 cells was also achieved by exogenous addition of TGFβ1 and the cells showed many of the characteristics of epithelial to mesenchymal transition (EMT) including a loss of ErbB3 expression, decreased expression of E-cadherin, increased vimentin expression, and reduced sensitivity to irreversible EGFR TKI. Activation of IR/IGF-IR was observed in the TGFβ1 treated H1975 cells. The mesenchymal-like NSCLC cells showed increased migration and upregulation of intrinsic signaling pathways controlling cell adhesion. The findings of receptor tyrosine kinase switching and deregulation of intrinsic signaling pathways suggest investigation of new therapeutic targets in NSCLC with mesenchymal phenotype.

Takeshi Shimamura Bio:
Since 2008, Dr. Shimamura has been an Instructor of Medicine at Dana-Farber Cancer Institute - Harvard Medical School, where his research focuses on the molecular biology - signal transduction of cancer and preclinical characterization of small molecule drugs for cancer. Projects that he is currently involved with at Dana-Farber Cancer Institute include: understanding interactions between epithelial to mesenchymal transition (EMT) and drug resistance, studying cell signaling network essential for metastasis in non-small cell lung cancer (NSCLC) with KRAS mutations, and preclinical characterization of non-geldanamycin Hsp90 inhibitors. His current research program is supported by Claudia Adams Barr program in innovative basic cancer research from Dana-Farber Cancer Institute.

As a postdoctoral fellow at Dana-Farber Cancer Institute, Dr. Shimamura contributed to the discovery of the therapeutic potential of Hsp90 inhibition in lung cancer. He identified that NSCLC cell lines harboring epidermal growth factor receptor (EGFR) mutations are exquisitely sensitive to Hsp90 inhibitors. Patient with NSCLC harboring EGFR mutations are routinely treated with EGFR inhibitors, which initially produce dramatic responses. Unfortunately, patients almost always succumb to acquired resistance to these inhibitors, due to the emergence of a secondary T790M mutation on EGFR. A proposed solution was the use of irreversible EGFR inhibitors to overcome T790M mutation. Dr. Shimamura has shown that irreversible inhibitors have substantial potency limitations against EGFR with T790M. He has demonstrated that Hsp90 inhibitor and EGFR irreversible inhibitor combined with rapamycin could overcome the acquired resistance mediated by the emergence of  T790M. These findings have been translated into clinical trials in patients with EGFR mutant NSCLC that are currently ongoing. His continued investigation of Hsp90 inhibition in lung cancer has evolved into a full project in the Dana-Farber/Harvard Cancer Center Lung specialized program of research excellence (SPORE), sponsored by NCI.

He received his Ph.D. at Western Michigan University (Kalamazoo, MI), where he studied roles of NF-kB in PDGF-BB mediated cellular transformation.

Patrick R. Sosnay, M.D.
Instructor, Pulmonary and Critical Care Medicine
Johns Hopkins Medicine

Presentation:  Genetic Testing for Screening and Diagnosis of Cystic Fibrosis


Abstract:
Cystic Fibrosis (CF) is the most common recessive lethal disorder in Caucasians. It is caused by mutations in the CF Transmembrane Conductance Regulator (CFTR), of which there have been over 1600 different variants characterized. Advances in DNA diagnostic technology allow greater coverage of that diversity, but creates the scenario where our ability to detect CFTR variants outpaces the knowledge to interpret the disease liability of those variants. CF is a model disease to employ newborn and carrier screening as the disease has a high prevalence, the consequences of the disease are severe, and because early diagnosis allows therapeutic interventions to potentially alleviate these consequences. CF also can be an instructive disease to analyze how the disease liability of genetic susceptibility variants is analyzed.

This talk will include an introduction to the clinical features of CF, and how these features are used for traditional CF diagnosis. We will also discuss the past, present, and future of CF genetic testing. The challenge created by CFTR variants of uncertain clinical significance pertains to both diagnosis and screening. Therefore, CF represents a unique opportunity to develop an algorithm to address and investigate uncertain genetic variants that will be applicable to a broad variety of diseases with genetic susceptibility.

Patrick R. Sosnay Bio:
Dr. Sosnay is an Instructor at the Johns Hopkins University, where he works under the mentorship of Garry Cutting M.D in the McKusick Nathans Institute of Genetic Medicine. Dr. Sosnay attended undergraduate and medical school at the University of Wisconsin. He then moved to Johns Hopkins University in Baltimore, Maryland where he has stayed for medicine residency, chief resident year, fellowship in Pulmonary and Critical Care Medicine, and as faculty. His research has focused on the cellular and clinical consequences of CFTR mutations. With Dr. Cutting and international CF collaborators, Dr. Sosnay has assembled the Clinical and Functional Translation of CFTR (CFTR2) project. This project uses clinical characteristics, laboratory-based testing, and predictive algorithms to assess the disease liability of a broad range of CFTR mutations and to communicate this information to the CF scientific community, clinical providers, and the general public. Dr. Sosnay is also an attending in the adult CF clinic.

Mary Sue Leffell, Ph.D., Diplomate, ABMLI, ABHI
Professor of Medicine and Director
Johns Hopkins Immunogenetics Laboratory

Presentation: Applications of Luminex® Technology in Histocompatibility and Immunogenetics

Abstract:
Definition or “typing” of antigens and alleles of the human major histocompatibility gene complex, the HLA system, is performed as an integral component of solid organ and hematopoietic stem cell transplantation. As many transplant candidates become pre-sensitized to HLA antigens and form antibodies that become a barrier to transplantation, antibody detection and identification are also critical parts of histocompatibility evaluations. HLA typing is confounded by the number of HLA alleles: the HLA system is the single most polymorphic human genetic system with over 4000 defined alleles. Definition of the specificity of antibodies to HLA antigens becomes difficult when patients are highly sensitized with antibodies directed toward numerous antigens. The ability to incorporate multiple sets of oligonucleotide probes greatly facilitates HLA typing through a reverse hybridization technique. Similarly, coupling of purified HLA antigens to Luminex beads provides a highly specific and sensitive method for identification of HLA specific antibodies. Because of these features, Luminex®technology is one of the most widely used platforms in histocompatibility laboratories.

M. Sue Leffell Bio:
Dr. Mary S. (Sue) Leffell is  Professor of Medicine at the Johns Hopkins University School of Medicine, Professor of Molecular Microbiology and Immunology at the Johns Hopkins University Bloomberg School of Public Health, and a Director of the Johns Hopkins Immunogenetics Laboratory.  Dr. Leffell graduated summa cum laude from the University of Tennessee, Knoxville and, as a Woodrow Wilson fellow, received her doctorate in immunology from the University of North Carolina, Chapel Hill, where she also completed a fellowship in clinical laboratory immunology. In addition to medical laboratory immunology, her specialty training is in histocompatibility and immunogenetics. She has been an active participant in clinical transplantation in the U.S., having served two terms on the Board of Directors of the national transplant network, the United Network for Organ Sharing (UNOS) and as Chair of the UNOS Histocompatibility Committee and as a member of the Kidney Transplantation and the Kidney Allocation Review committees. She recently completed her term on the Advisory Committee on Organ Transplantation to the U.S. Secretary of Health and Human Services.  She is a past President of the American Society for Histocompatibility and Immunogenetics and served as a member of the scientific advisory committee for the 15^th International HIstocompatibility and Immunogenetics Workshop and Conference. She serves on the editorial boards of Human Immunology, Transplantation, and previously on the board of Clinical and Diagnostic Immunology. Her research interests are in immunogenetics and transplantation immunology with a recent focus on the impact of HLA and KIR alleles in hematopoietic stem cell transplantation. She is a member of the American Association of Immunologists, the American Society of Transplantation, the Association of Medical Laboratory Immunologists, and the International Transplantation Society.

Peter A. Zimmerman, Ph.D.
Professor, The Center fro Global Health and Diseases
Case Western University

Presentation: Multiplex Diagnosis Contributes to Malaria Elimination

Abstract:
Malaria is endemic in 109 countries in the world today. In these countries over 3 billion people are at risk for developing malaria, 300 million experience clinical malaria, and over 1 million people die from malarial disease annually.  In Africa, economists have estimated that direct losses (e.g. illness, treatment, premature death) reach US$ 12 billion annually and many times more than that in lost economic growth. At all stages of the Global Malaria Action Plan diagnosis plays an important role in progress toward elimination of malaria as a threat to public health. Multiplex ligase detection reaction/fluorescent microsphere based assays (LDR-FMA; Luminex® FlexMAP™ microsphere technology) strategies incorporating Luminex reagents and analyzers have been developed to perform parasite diagnosis in human and mosquito phases of the malaria lifecycle. Species-level assays differentiate all human as well as additional non-human primate malaria parasite species. As malaria control efforts have resulted in a worldwide increase in resistance to drugs used to treat the disease, we have developed additional assays to identify SNPs associated with resistance in Plasmodium falciparum and P. vivax. These multiplex diagnostic assays are essential tools for defining the scope of baseline malaria complexity and assessing the impact of malaria control programs, from ‘scale-up for impact’, ‘sustained control’ and ‘elimination’ of malaria from endemic regions around the world.


Peter A. Zimmerman Bio:
Pete Zimmerman received his Ph.D. from the Case Western Reserve University Department of Biology in 1992. His Ph.D. thesis investigated the association between strains of the parasitic nematode Onchocerca volvulus with blinding or less severe pathogenesis. Dr. Zimmerman received a National Research Council Associateship award to continue his post-doctoral education at the National Institutes of Health with Thomas Nutman in the Laboratory of Parasitic Diseases. Dr. Zimmerman's studies on genetic factors influencing susceptibility to infectious disease at the NIH culminated in identification of human genetic polymorphism conferring resistance to HIV-1 infection and delayed progression to AIDS. Dr. Zimmerman joined the CWRU faculty in 1997.  Since then his laboratory has developed multiplex diagnostic platforms for diagnosing complex infections by numerous malaria parasite species, drug resistant strains and antigenic variation of malaria vaccine candidates. Through Dr. Zimmerman’s epidemiologic studies Luminex-based diagnostic methods have been transferred to malaria-endemic sites in Papua New Guinea and Africa.     

Jill Simonetti
Newborn Screening Scientist
Minnesota Department of Health

Presentation: Newborn Screening in Minnesota and Benefits of Multiplexing

Abstract:
Since 1965, the Minnesota Department of Health’s Newborn Screening Program has screened Minnesota newborns soon after birth to see if they are at risk for hidden, rare disorders. Without treatment, these disorders can lead to illness, physical disability, mental retardation, or death. Medications or changes in diet help prevent most health problems caused by disorders that are identified through newborn screening. The Minnesota Department of Health together with hospitals, laboratories, and medical professionals across the state screen for hearing loss and 53 disorders that may affect an infant’s metabolism, endocrine system, blood, breathing, hearing, or digestion. Each year the lives of approximately 400 children are saved or improved by newborn screening in Minnesota.

The Minnesota Newborn Screening Laboratory is participating in a study of a new multiplex assay developed by Luminex. The purpose of the study is to verify the performance of the assay and generate data for Luminex’s 510K submission. The new Luminex assay simultaneously screens for Congenital Adrenal Hyperplasia (CAH), Congenital Hypothyroidism (CH), and Cystic Fibrosis (CF) from a single dried blood spot punch. Minnesota currently screens for all 3 of these disorders using 3 different assays and 3 different punches.

The Luminex assay uses bead-based technology and may have increased sensitivity and specificity for detection of CAH, CH and CF over the current flouroimmunoassays. Improved sensitivity and specificity means fewer false positives and false negatives. Additional benefits of this new multiplex assay include: reduced specimen quantity, an additional biomarker for hypothyroid screening, possible performance improvement over our current 3 assays, possible replacement of our current testing platform, and the opportunity to provide feedback from a state screening lab that may improve the assay prior to FDA approval.

This presentation will give an overview of newborn screening in Minnesota including a summary of our experiences with the Luminex system and why multiplexing is advantageous in newborn screening.

Jill Simonetti Bio:
Jill Simonetti is a Scientist in the Newborn Screening Laboratory at the Minnesota Department of Health. She has held this position since 2002. She also serves as the QC/QA officer for the laboratory. Jill is a team leader in the lab and responsible for training new personnel. She has an advanced level of experience performing various high-throughput laboratory assays, including PCR, LC/MSMS, HPLC, DNA extraction and purification, ELISA assays, isoelectric focusing, flourometry, and flow cytometry. Jill has expertise in developing, writing, and presenting technical publications, validations, and SOPs. She holds a Bachelor’s Degree in Biology

Laurie M. Clotilde-Bollinger, Ph.D.
Research Biologist
USDA Agricultural Research Service

Presentation:  Detection and Recovery of Shiga Toxins and Escherichia coli O157

Abstract:
Shiga toxin-producing Escherichia coli (STEC) are among the most costly foodborne pathogens.  In the United States, recent annual cost estimates for acute care ranged from $1 to $2 billion.  These were based on the assumption that E. coli O157:H7 led to 73,000 illnesses and 61 deaths each year.  The pathogenesis of these infections depends on the production of Shiga toxin 1 (Stx1) and/or Shiga toxin 2 (Stx2).  Currently, there are no commercially available kits capable of detecting Stx1, Stx2, and O157 lipopolysaccharides (LPS) simultaneously.  As STEC strains producing Stx1, Stx2, or both toxins have often been linked to outbreaks of human illnesses and most of these are traced to consumption of E. coli O157-contaminated foods, this study focused on this specific STEC serotype as well as the main virulence factors.  Here, we developed a Luminex-based immunoassay to screen for Stx1, Stx2, and E. coli O157 LPS simultaneously in spiked foods.  Using minimal sample preparation, we were able to detect the three analytes simultaneously and our results showed the same specificity and sensitivity as ones obtained from testing pure STEC cultures.  Conventional sandwich ELISA using the same antibodies was not as sensitive.  Our newly developed Luminex-based immunoassay will serve as a milestone for developing a multiplex immunoassay for additional foodborne pathogens.

Laurie M. Clotilde-Bollinger Bio:
Dr. Laurie M. Clotilde grew up in Belgium and came to the United States in 1999 where she completed her education at the University of Nevada, Reno.  In 2004, she received a M.S. in Animal Sciences followed by a Ph.D. in Cellular and Molecular Biology in 2008.  Her graduate work focused on exploring the factors affecting the prevalence of Shiga toxin-producing Escherichia coli (STEC) in cattle and identifying the molecular aspects of the recovered E. coli O157 and non-O157 STEC isolates.  Dr. Clotilde also has extensive experience in culturing both E. coli O157 and non-O157 STEC in fecal, food, and environmental samples.  For the past year and a half, she has been a Research Biologist (Post-doctorate) in the Foodborne Contaminants Research Unit of the USDA-ARS.  With her supervisor (Dr. J. Mark Carter), she is currently collaborating with the FDA-CFSAN (San Francisco District Office, CA) to develop and optimize a Luminex-based immunoassay for detection of toxic E. coli markers (Shiga toxins 1 and 2) and identification of E. coli O157 in various food matrices.

P. Scott White, Ph.D.
Acting Program Manager for Chem/Bio/Med Countermeasures
Los Alamos National Laboratory

Presentation:  Multiplex DNA/RNA-based Assays for Detection, Characterization, and Genotyping

Abstract:
Nucleic acid-based assays are ideal for pathogen detection and diagnostics, as well as for genotyping due to their high specificity, the ease with which they can be configured to detect almost any target, their requirement for minimal quantities of sample (high sensitivity), and their ability to be automated for high throughput applications. In addition, recent advances allow such assays to be configured in a multiplex format, enabling simultaneous screening for multiple genetic targets.  Examples of such applications include surveillance for multiple pathogens using multiple genomic targets for each, performing identification and genotyping on many sites simultaneously, or to perform extensive genetic characterization of samples of known identity.  There is a huge cost advantage to performing multiplex that allows even more targets to be interrogated, whereas cost is a common limiting factor for singleplex assays.

One challenge faced in developing assays to address such demanding applications is the lack of suitable and robust assays that perform well in multiplex. Although there are several analysis platforms capable of reading many different assay results, their utility is limited by the use of assays that do not perform well in a multiplex. In many cases, assays designed for singleplex formats are applied to multiplex analysis instruments with poor results. We have developed an assay, called Multiplex Oligonucleotide Ligation-PCR (MOL-PCR) that was designed for multiplex, and uses a flow cytometer for analysis. The assay uses generic or universal components where possible, which add much needed flexibility in design and reconfiguration, whereas specificity is conferred by careful nucleic acid target selection and probe design. Results of several Luminex-based MOL-PCR assays that address a diverse collection of detection and surveillance applications will be presented.

P. Scott White Bio:
Dr. White is currently a Scientist and Team Leader for Applications and Analysis, in the Biosecurity and Public Health Group, Bioscience Division at Los Alamos National Laboratory (LANL). His latest work involves the development of multiplex DNA/RNA-based assays for the detection and characterization of pathogenic microorganisms. Recent and current projects are to develop assays for detection of biothreat agents, to determine antibiotic resistance of biothreat agents using single nucleotide polymorphisms and detection (presence/absence) targets, and to develop detection and strain identification assays for citrus pathogens, among others. Dr. White has been involved in technology development for a number of years, and holds US and foreign patents on multiplex nucleic acid-based assays that utilize flow cytometry as the analysis platform. His is currently working as the Program Manger for Chemical, Biological, and Medical Countermeasures for the Homeland Security Program Office at LANL. Prior to his work on pathogen detection and characterization assays, Dr. White worked on technology development for the Human Genome Project, and has also used DNA and RNA sequence data to estimate the phylogenetic relationships among crocodilia and plant RNA viruses. Dr. White received his Ph.D. in Biology from Texas Tech University in 1992, did postdoctoral work at the Samuel Roberts Noble Foundation, and has worked at Los Alamos National Laboratory since 1995.

Aristidis Veves, M.D., D.S.C.
Research Director, Microcirculation Lab and
Joslin-Beth Israel Deaconess Foot Center
Associate Professor, Harvard Medical School

Presentation: Employing Luminex Technology to Identify Vascular and Metabolic Changes in the Diabetic Foot

Abstract:
We have employed a Luminex 200 apparatus (Luminex Corporation Austin, TX) and Millipore multiplex immunoassay panels (Millipore Corporation, Chicago, IL) to measure various cytokines and growth factors in ongoing studies in our unit.  Our results indicate: 1.) a reduction in the EGF, FGF, IFNg, MIP1a and MIP1b serum levels in diabetic patients who develop foot ulceration and an increase in the PDGA-AA, FGF, leptin, G-CSF, GRO. MCP-1, MMP-9 and tPA-1 serum levels in diabetic patients who fail to heal their ulcers. 2.) neuropathic diabetic patients have higher serum levels of PDGF AA/BB, RANTES, leptin, osteoprotegerin, G-CSF, sE-Selectin, sICAM, sVCAM, CRP, TNFα and fibrinogen.  Patients with painful neuropathy have higher cICAM-1 and CRP levels when compared to painless neuropathy, 3.) Obese non-diabetic subjects have higher VEGF, obese diabetic patients have higher TNFa while both obese non-diabetic and diabetic subjects have higher CRP.  Insulin measurements using this technology have a very satisfactory correlation with standard measurements (r=0.86).

Aristidis Veves Bio:
Dr. Aristidis Veves is the Research Director of the Joslin-Beth Israel Deaconess Foot Center and of the Microcirculation Lab, and Associate Professor at Harvard Medical School.  He received his M.D. from the Medical School, Aristotelion / University of Thessaloniki (Greece), his M.Sc. from the Faculty of Medicine, University of Manchester (U.K.), and his D.Sc. from Athens Medical School (Greece).  Dr. Veves did his internship in General Medicine at Athens Naval Hospital (Greece), and completed his residency in General Medicine at Tsaggari General District Hospital (Athens, Greece).  He then served as Hon. Senior House Officer in the University Department of Medicine, Diabetes and Endocrinology at Manchester Royal Infirmary (U.K.).  Most recently, he was a Research Fellow at Deaconess – Joslin Foot Center, Deaconess Hospital (Boston, MA).

Dr. Veves’ main academic interest is the role of neuropathy and vascular disease in the development of diabetic foot problems.  Dr. Veves is also involved with evaluating the endothelial function of the brachial artery, as well as other studies involving endothelial function.  In summary, the main effort of Dr. Veves’ research is to develop techniques which will identify patients at risk of developing diabetic foot problems at the early stages and to develop therapeutic strategies which will prevent or slow the progression of this condition. In addition, Dr. Veves is also interested in the etiology of diabetic foot problems and the pathophysiology of wound healing in diabetes.

Dr. Veves’ areas of expertise include diabetic neuropathy, diabetic foot problems, and vascular reactivity.

Imran H. Khan, Ph.D.
Associate Director, Clinical Proteomics Core
University of California, Davis

Presentation:  New Paradigm in Tuberculosis Serodetection: Computational Analysis to Classify Profiles of anti-M. tb. Antibodies Detected by Multiplex Microbead Array

Gary Procop, M.D., M.S.
Chairman of the Department of Clinical Pathology
Cleveland Clinic Foundation

Presentation: The Multiplex Nature of Diagnostics for Infectious Diseases:  Making the Leap from Culture to Molecular - What Will the FDA Do?

Abstract:
The diagnosis of infectious diseases is a complex process since a variety of different microorganisms can cause the same types of diseases and the different signs and symptoms that can be produced within the human body are limited. The past hundred years has seen the refinement of traditional laboratory methods, such as culture and morphologic analysis, which are useful for identifying the particular cause of an infection. Unfortunately, these methods are often time-consuming, require human expertise, and remain lacking in sensitivity and specificity in many instances. Multiplex and broad-range PCR assays with post-amplification analysis offer the possibility to improve the time-to-diagnosis, and in some instances perhaps improve sensitivity and specificity. Another advantage to employing molecular approaches for a variety of conditions is that a single skill set is needed to perform testing rather than a number of different skills. The design and implementation of these assays in a financially strained healthcare system will be challenging, as will the review and approval of these assays by the Food and Drug Administration (FDA). We will explore instances wherein molecular diagnostic assays have successfully replaced traditional methods, as well as areas wherein molecular diagnostics hold promise and the potential corresponding challenges of FDA approval.

Gary Procop Bio:
Gary W. Procop, MD, MS is Chairman of the Department of Molecular Pathology, Section Head of Molecular Microbiology, and Director of Mycology, and Parasitology at the Cleveland Clinic.  He completed Anatomic and Clinical Pathology training at Duke University Medical Center and a Clinical Microbiology Fellowship at the Mayo Clinic.  He is a diplomat of the American Board of Pathology in Anatomic and Clinical Pathology, and Medical Microbiology. He is a Fellow of the American Academy of Microbiology, the College of American Pathologists, the American Society for Clinical Pathology, the Infectious Diseases Society of America, and the Royal Society of Tropical Medicine and Hygiene.  He has given more than 375 scientific presentations, and has 124 published manuscripts, 25 chapters, and one book to his credit.  He is a Trustee of the American Board of Pathology, and Chair of the Microbiology Test Development Committee for the Board.  He is also a Member of the Board of the American College of Microbiology.  He holds the Vice Chair position on both the Public Affairs Committee and the Microbiology Resource Committee for the College of American Pathologists.  His primary interests are the practical applications of molecular diagnostic methods for the diagnosis and treatment of infections; infectious disease pathology; mycology and parasitology.  His main hobby is sailing, and he holds certifications in Basic Keelboat Sailing, Basic and Intermediate Coastal Cruising, and Coastal and Celestial Navigation.   

Christine Ginocchio, Ph.D., M.T. (A.S.C.P.)
Director, Microbiology, Virology, and Molecular Diagnostics
North Shore LIJ Health System Laboratories

Presentation:  Comprehensive Molecular Diagnostics as a Critical Component of a Respiratory Virus Surveillance Program

Abstract:
Disease recognition within a community can be triggered by several key indicators, including syndromic surveillance when care is rendered in an Emergency Department setting or by comprehensive laboratory-based diagnostic results. The value of each component is unique and significant.  However, in combination, syndromic and laboratory surveillance offer the potential to rapidly identify unusual infectious disease related events.

The timely identification of respiratory viral infections is of tremendous importance for diagnosis, infection control, epidemiologic and surveillance studies. Monitoring seasonal viral patterns over many years provides baseline information by which both health care systems and public health officials can detect unusual changes in patterns or unexpected outbreaks.  The North Shore-LIJ Health System Laboratories provide diagnostic testing for 13 hospitals and 500 private physician offices, which geographically extend from NY City to the end of Long Island, covering approximately a 6.5 million persons population base. Therefore, due to the extent of the patient coverage and the comprehensive diagnostic testing offered by the laboratories, emerging respiratory infectious diseases or changes in seasonal patterns are readily and rapidly identified.  The power and scope of a comprehensive diagnostic program is best exemplified by the identification, management, and monitoring of the NY City outbreak of pandemic 2009 novel influenza A H1N1.  This lecture will discuss the role of comprehensive respiratory virus testing in the setting of ongoing diagnostic and epidemiology surveillance programs.


Christine Ginocchio Bio:
Dr. Ginocchio received her bachelor’s degree (Summa Cum Laude) in Medical Technology from St. John’s University, NY (1973), completed a 1 year clinical laboratory internship at Nassau County Medical Center, East Meadow, NY and worked as a Medical Technologist in Microbiology at St. Charles Hospital, Pt Jefferson, NY until 1993.  She completed her Ph. D. (highest distinction and President’s Award for Outstanding Research) in Molecular Microbiology and Genetics from the Department of Microbiology and Genetics, School of Medicine, State University of New York, Stony Brook, NY in 1993. Her thesis research, under the mentorship of Jorge Galan, Ph. D., was on the genetics and cell biology of Salmonella pathogenesis for which she received a Sarber Fellowship from the ASM.  Dr. Ginocchio holds a NY State Certificate of Qualification as a Laboratory Director and has been the Director of Clinical Microbiology/Virology, Molecular Diagnostics and HIV Testing services for the North Shore-Long Island Jewish Health System Laboratories, NY since 1994. The NS-LIJ Laboratory performs routine and reference diagnostic testing for 11 hospitals, including 3 university tertiary care facilities, 50 nursing homes, clinics and over 500 physician offices. 

Dr Ginocchio is a member of the North Shore University Hospital Medical Staff, Department of Medicine and Division of Infectious Diseases. Her responsibilities include numerous hospital and laboratory committees (Infection Control, Pharmacy and Therapeutics, Bio-preparedness, Laboratory Medical Advisory Board, Methods Committees for Immunology, Microbiology and Molecular Diagnostics [Chair]).  She was a recipient of the George M. Jaffin Department of Medicine Scholarly Activity Award.  Dr Ginocchio is responsible for the NS-LIJ Microbiology staff education for adult and pediatric infectious disease fellows and pathology residents. She is a member of the LCME Committee for the development of a new medical school in collaboration with Hofstra University, NY. Her faculty appointment (15 yrs) is at the State University of New York at Stony Brook, Department of Microbiology and Genetics, School of Medicine, where she directs the Medical School Microbiology Laboratory Course.   Dr. Ginocchio is a member of ASM (Laboratory Practices Committee), PASCV (former counselor), ASCP, AMP and the CAP Microbiology Resource Committee.  Dr. Ginocchio is the Co-Editor in Chief for the Journal of Clinical Virology, a Virology Section Editor, Manual of Clinical Microbiology 10^th Edition and on the Editorial Board for Clinical Microbiology Reviews.

Dr Ginocchio’s area of extra-murally funded research (funding exceeds $2.67 million) have included HIV, CMV, respiratory viruses, HPV and molecular diagnostics for Infectious Diseases. She currently receives government funding from the NIH/NIAID (Detection of Blood Stream Pathogens), Department of Defense (Respiratory Virus Surveillance) and the NY State Department of Health (MRSA).  She has been the principal investigator for more than 30 clinical trial studies, which include 13 studies of in vitro diagnostic devices for FDA clearance.  She has been an invited speaker at over 100 national and international conferences, has over 140 articles and abstracts and 2 clinical virology book chapters.  Dr. Ginocchio is currently a member of five International Advisory Panels for Diagnostic Biotechnology Companies and is the Chair of the Compass Group Molecular Diagnostic sub-committee.

Jean Louis Merlin, PharmD, Ph.D.
Professor in Cellular Oncology
Alexis Vautrin Cancer Center

Presentation:  Phosphoprotein Array Assay for Characterization of Human Tyrosine Kinase Receptors Downstream Signaling Functionality as Response Predictive Marker to Targeted Therapy in Translational Oncology Research Programs

Abstract:
The functionality of human epidermal growth factor receptors (HER) downstream signaling kinases have major consequences on tumor response to anti-HER targeted therapy. In our research project, Luminex® phosphoprotein array (PA) was used to analyze the expression of p-MEK, p-ERK1/2, p-P38MAPK, p-AKT, p-GSK3, p-P70S6K in cell lines (preclinical models) and in frozen biopsies taken at diagnosis from breast, head and neck and colorectal cancer patients.

In preclinical transgenic cells, PA was found to be very useful in understanding the role of key-determinants of the cellular response to anti-HER directed drugs or stategies (Cancer Gene Ther, 2009). In patients, using optimized standard operating procedures (freezing delay, specimen quality, protein extraction), PA was cross-validated with western blot analysis in 49 breast cancer specimens and the two methods showed clinical comparability (Clin Chem, 2009). In all tumor types, great variations (up to several-hundred folds) in phosphoprotein expression were observed among the specimens revealing high-range inter-individual variations. In metastatic colorectal cancer, PA results were confronted to KRAS mutation, used today as response predictive marker to anti-EGFR monoclonal antibodies. KRAS and p-MEK were identified as two independent prognostic markers of patients treated by cetuximab. In wild type KRAS tumors, p-MEK and p-P70S6K overexpression were found to be associated with significant lower progression-free survival showing that PA could be used for clinical response prediction (Int J Cancer, 2010).

As a whole, our results validate the use of PA for single-step analysis of signaling pathways functionality in translational oncology research programs. In either preclinical or clinical research programs, PA is a potent tool for evaluation and validation of molecular determinants / clinical response predictive markers of cellular / tumor response to anti-HER directed targeted therapy.

Jean Louis Merlin Bio:
Jean-Louis Merlin received his PharmD at the University of Lille in 1984 the his PhD In Biomedical Engineering at the University of Nancy in 1992. He was appointed Head of the Research Laboratory in 1988 then Head of the Tumor Biology Unit in 2005 at Alexis Vautrin Cancer Center in Nancy. Jean-Louis Merlin became Professor in Cellular Oncology at the Faculty of Pharmacy of the University of Nancy in 2004.

Jean-Louis Merlin is member of executive boards of several research groups of the French National Federation of Cancer Centers (FNCLCC) and European Organization for Research and Treatment of Cancer (EORTC) research groups and is member of scientific boards of several pharmaceutical companies.

His research field of activity has been devoted to identification and circumvention of markers of resistance to anticancer treatments, chemotherapy, radiation therapy and photodynamic therapy and molecular targeted therapy. His activity in the hospital is mainly devoted to the identification, validation and clinical application of molecular markers for response prediction to molecular targeted anticancer therapies namely therapeutic monoclonal antibodies and kinase inhibitors.

In 2008, Jean-Louis Merlin participated to the creation of a research group in Oncology named SiGReTO, Signaling, Genomics and Translational Research in Oncology gathering molecular biologists, radiobiologists, cellular oncologists, physiologists, pathologists, medical doctors involved in clinical trials in a research platform including in vitro and in vivo imaging (intravital microscopy, microimager, animal MRI and PET scanning, …) core facilities and clinical research hospital units.

Jean-Louis Merlin has participated to more than 80 international peer-reviewed papers and presented his work in many international oncology meetings.

Matthew Myles, D.V.M., Ph.D., DACLAM
Research Animal Diagnostic Laboratory
Veterinary Pathology
College of Veterinary Medicine
University of Missouri

Presentation:  Improving the Overall Accuracy of Serological Diagnostic Assays

Abstract:
The test characteristics, sensitivity, specificity, positive predictive value and negative predictive value, are important ways to express the usefulness of a diagnostic assay. A test’s sensitivity and specificity are the probabilities that the test result will be positive or negative in the presence or absence of disease, respectively; whereas, the predictive values are the probabilities that a patient will or will not have a disease given a positive or negative test result, respectively. Thus, a test’s sensitivity and specificity are properties of the test and should be consistent for a given test when applied with comparable methodologies. Predictive values, although related to sensitivity and specificity, vary with the prevalence of the disease being evaluated and, all things being equal, the lower the prevalence of disease, the lower the predictive value of a positive result. Surveillance of rodents used in biomedical research for adventitious infections is critical for validating the reliability of research results. Previous reports of laboratory rodent disease surveillance have demonstrated that the prevalence of most adventitious infections in rodent colonies is less than two-percent. Thus, given the low true positive rate of adventitious agents in rodent colonies and the concomitantly low positive predictive value of tests used to screen for those agents, a positive assay result for an agent of low prevalence is likely to be a false positive. To address this issue we hypothesized that incorporating two serological assays for diagnosing a single infectious agent would improve the positive predictive value and overall accuracy of diagnosing adventitious infections in rodents. To test this hypothesis, we used a single data set (n=1879) comprised of serum samples from murine norovirus (MNV) positive and negative mice randomly split into a training dataset (n=628) and a test dataset (n=1269). These samples were evaluated against a Multiplex Fluorescent Immunoassay (MFI) for simultaneous detection of antibodies to MNV-whole virus (MNVwv) and MNVVP1/2.   We fit the following three multivariate logistic regression models using the training data: Model 1, MNVwv as only predictor; Model 2, MNVVP1/2 as only predictor; and Model 3: MNVwv and MNVVP1/2 both as predictors. The best cutoff point was determined for each model and then prediction results for the test data were calculated using the developed model. Assuming prevalence of disease is 29%, the positive predictive values were: Model 1, 94.45%; Model 2, 97.47%; and Model 3, 98.04%. Thus, two assays used to diagnose MNV infection improve the positive predictive value and overall diagnostic accuracy as compared to either test alone.

Matthew Myles Bio:
Matthew Myles is Associate Director of Serology Services in the Research Animal Diagnostic Laboratory within the Department of Veterinary Pathobiology at the University of Missouri. He completed his bachelor’s and DVM degrees at Colorado State University and earned a PhD in Veterinary Pathobiology from the University of Missouri. He completed Laboratory Animal Medicine training at the University of Missouri and is a diplomate of the American College of Laboratory Animal Medicine.

Dr. Myles research interests are in the area of host-microbe interactions. He has used Helicobacter hepaticus-induced typhlitis in mice, a model of bacterial driven chronic intestinal inflammation that closely mimics human Crohn's disease, as a tool for studying the immune signals that initiate mucosal inflammation as well as those that perpetuate disease. Projects, ongoing in his laboratory, are focused on defining the gene expression profile in early and late typhlitis, determining the role of mucosal epithelial cells in regulating or enhancing the inflammatory response, and identifying key proinflammatory and immune-regulatory mediators.