Clinical Research Scientist, Cellular and Molecular Biologist/Biochemist

Clinical Research Scientist

ANTHONY RAFFO, PhD, MS

Cellular and Molecular Biologist/Biochemist

Summary

v  10+ years’ experience as cell and molecular biology research scientist with strong focus on pre-clinical and clinical assay development and technology optimization

 

v  10+ years’ experience working as a Scientist in the study of human diseases with emphasis on cellular and animal models of pharmacokinetics and pharmacodynamics studies

 

v  10+ years’ experience in drug discovery/clinical development with molecular mechanistic toxicology assays

 

v  10+ years’ experience with immunological techniques for protein detection, expression and quantitation, including ELISA, Western blot analysis, RIA, FACS and FISH.  Also including assessment of comparative antibody quality using competitive binding studies.

 

v  5 years’ experience in Molecular Profiling and biomarker identification/detection technologies (i.e. mRNA microarray profiling, microRNA profiling, biomarker diagnostic assay development, DNA genotyping, sequencing [Sanger and NGS platforms (PGM and MiSeq)] )

 

v  7 years experience in confocal microscopy and PET imaging of cellular and organ functioning, with 2 years clinical imaging; with a focus on oncology and tumor evaluation in therapy studies.

 

v  10+ years’ experience with cell culture of human as well as insect, rodent, yeast and bacterial cultures with flow-cytometric analysis (FACS) of ligand binding and signal pathway activation analyses

 

v  15+ years’ experience with Computer programs dealing with office, statistical and bioinformatics tools with data-mining, writing and submission/publication of peer reviewed articles, proposals and SOPs; with several presentations in large peer scientific conferences (AACR, AUA, etc)

 

v  15+ years of  lab manager/supervisor in bio-medical pre-clinical assay development and retrospective clinical research labs with 1 year experience in a GMP environment

 

v  2 years’ experience in automated liquid and plate handling equipment in bioassay development

 

v  Lab managerial and organizational experience with equipment operation and maintenance and supplies procurement and stocking; support personnel supervisor and instructor

 

ü  Independent investigator with a strong belief in internal team and external investigator collaboration to share ideas and expertise.   Firm belief that new concepts and novel interpretations of long established principles, coupled with innovative technology with a multi-discipline approach, can lead to the resolution of difficult proposals, in order to efficiently complete a specific project (one that will hopefully benefit human health)

 

 

 

 

Contact Information                                                         

                    Current address:

 

 

 

                       Anthony J Raffo                                    

                       7316  10th Ave                         

tonyraffo@yahoo.com

 

    

                    Brooklyn, NY                                                            917-402-6047

 

 

 

 

Professional Experience

 

 

Clinical Development Laboratory-Genomics, Merck, Rahway, NJ                                    Feb. 2012 to April 2013

Biochemist III/ Genomics Staff Scientist 

I was recently a contract scientist, at Merck Clinical Development Laboratories, working at Clinical Assay Development in Genomic Profiling and Gene Expression Analysis.  I was the lead scientist in the preparation and establishment of assays, procedures and protocols, adhering to GLP compliant practices, prior to regulatory approval, used in the clinical trials of potential therapeutics for oncologic, immunologic and infectious diseases in human treatment applications.  I was the lead scientist on several genomic based projects utilizing next generation sequencing, genotyping with microarrays and gene expression analysis based on qRT-PCR and microarrays.  I organized Merck’s efforts to establish several platforms of next generation sequencing at Merck, CDL-G, with the installation of MiSeq and PGM instruments and am scientific lead on a project to utilize the SMART instrument technology.  One of my projects was to develop and validate a genotyping assay for several potentially key single nucleotide polymorphisms in a very important gene, whose protein is implicated as causal in coronary arterial disease and in an additional project I established criteria to test and validate several vendors, which Merck needs as outsourcing partners in patient gene expression analysis.

I also interacted closely with the Merck Biomarkers group in coordinating biomarker discovery toward the development of a clinical assay for a therapy treatment decision in breast/ovarian cancer therapy.  Here, I optimized a FFPE sample core needle biopsy assay for beta-catenin expression as a result of various siRNA therapy protocols.

 

National Income Life Insurance Company, NY, NY                                                 June 2011 - Feb 2012

Managing Agent

I was a managing agent for National Income Life Insurance Company (NILICO), which is the largest provider of union based supplemental Life and health insurance in the US.    I optimized and developed supplement life and health insurance strategies for New York City area union members. NILICO is an enthusiastic advocate of child protection and during my visits to union and several non-union families, I distributed NILICO’s child identification kits, which is a confidential means, kept solely by the parents of young children, to fingerprint and document a child’s physical characteristics in case that child is later found missing or abducted.

 

From June 2009 to June 2011, while job searching I dealt with several pressing family issues; I also was a volunteer tutor, teaching English to foreign students, and I took this time to learn the fundamentals of the newly developed next generation sequencing platforms as well as, proteomics, especially techniques of Column Chromatography and Mass Spectroscopy and the fundamental molecular mechanisms of innate immunity                                                                                                                                    

 

 

                                                                                                                                                                            Jul 2008 - Jun 2009

High Throughput Center for Cell Biology, SiRNA screening Core Facility, Dept of Cell Biology, Yale University    Orange, CT                                                                               

Associate Research Scientist

  • 1 year experience in High Throughput (HT) Bioassay confocal microscopy siRNA screening with HT real time RT-PCR validation.  384 well plates, robotic liquid handling and automated plate manipulation

 

  • Help to pioneer Yale West Campus (formally part of Bayer Corp.) with the establishment of the siRNA screening core center. 
  • Main role as to validate siRNA based knockdown with the use of HT qRT-PCR of several key biomarkers and antibody based imaging (HT confocal microscopy) analysis and protein western blot or ELISA quantitation of these biomarkers.
  • High throughput screening; 96 and 384 plate format; automated liquid and plate handling

 

 

                                                                                                                                    Sep 1995 - Jul 2008

Columbia University Medical Center    New York, NY                                                                                                                 

Associate Research Scientist

 

The Naomi Berrie Diabetes Center

  •  Image the loss of  insulin producing beta cells within the pancreas of experimental rats by Positron Emission Tomography (PET) scanning, induced by streptozotocin or spontaneously generated in a genetically engineered diabetic prone rat model,
  • Correlate the imaging data with the appearance of beta cell specific peptides within the serum of these rats as diabetes progresses using mass spectroscopy analysis
  • Quantitatively measure the loss of beta cell specific proteins and their mRNAs (using the beta cell specific neuronal biomarker VMAT2) within the regressed pancreas in these diabetic rats by Western blots, ELISAs and real-time PCR
  • Image and semi-quantitate the localization of several key proteins in the pancreas of these animals and in a rat glucose inducible insulin secreting cell line, by confocal fluorescent immunohistochemistry focusing on pancreatic specific biomarkers.
    • Molecular bio/biomarker assay development (including flow cytometry and RT-qPCR based)
    • Signal transduction pathways effecting growth, death, differentiation and senescence, with emphasis on oncology (specifically prostate, breast, melanoma, and lymphoma cancers), immunology and metabolic disorders (diabetes)

 

 

 

Department of Medicine, Division of Medical Oncology, Experimental Therapeutics

 

  • Determine the in situ and in vivo application of a bio-active peptide from the C and N terminus of the key cancer biomarker tumor suppressor protein p53, which can activate some mutant forms of p53 in breast, lung, brain and prostate cancer cell lines, inducing cell death.   These studies included limited DMPK analysis of absorption, metabolism and kinetics of peptide in cell line studies.  Since p53 is the most mutated of all human proteins to be found in human cancers, these studies potentially can lead to the development of biologic and small molecule therapeutics for a wide range of cancer types.  Optimization of peptide constructs were conducted to enhance peptide stability and promote cell uptake.

 

Award- American Association for Cancer Research- GlaxoWelcome Investigator Award 2000. Raffo, A.J., Drew, L., Kim, A.L., Brandt-Rauf, P.W., Petrylak, D.P., Do, T. and Fine, R.L.  Selective Induction of fas Mediated Apoptosis by p53 Peptide in Human Cancer Cells

 

  • Experience in writing proposals, SOP’S, protocols and articles, as well as presenting data and conclusions to large peer scientific audiences with an over whelming desire to learn as much as possible through these interactions and through a conscientious reading of relevant scientific articles and publications

 

 

  • Design and constructed insect vectors for the production and isolation of peptides from several regions of the human DNA repair biomarker protein XRCC-1.  Part of this project is to determine the three dimensional structure of these conserved regions and to correlate this structure with several polymorphisms found in the human population.  An additional part of this project is to determine how these polymorphisms affect the DNA repair function.  As part of this project I trained a master’s student in protein production and purification and I was also a consultant to a doctoral student in general molecular biology techniques and in transfections of human lung cancer cells with the XRCC-1 constructs. 
  • Presented evidence for a nuclear complex between the pro-apoptosis regulating protein, BAX, and the genome guardian/transcription activator protein, p53 and that this complex formation correlated with the timing of the onset of apoptosis.  This project utilized confocal microscopy with immune-labeling techniques and selective immune-precipitation protocols, plus various cell and enzyme based apoptosis detection and quantification assays.  Biomarker protein quantification was done by Western blot, Dotblot or ELISA technologies, as appropriate.

Award-American Association for Cancer Research-Intergen Young Investigator Award 1999. Raffo, A.J., Kim, A.L. and Fine, R.L.  Formation of nuclear Bax/p53 complexes are associated with apoptotic DNA fragmentation.

  • Therapeutic chemical and biologic agents efficacy studies and combinatorial chemotherapy; DMPK/ADME studies

 

  • Determine the efficacy of certain experimental drugs for the treatment of cancers in situ and in vivo , including toxicology (limited DMPK/ADME in situ studies) of these agents either alone or in combination with existing chemotherapy agents.  In vivo imaging of patient tumor therapy progression by PET and MRI for prostate cancer patients in order to evaluate experimental therapy protocols.

Award-American Urological Association. 2003 Moderator of the Poster Discussion Session on Prostate Cancer at the 2003 A.U.A. National Meeting, Chicago

  • skilled in various cellular diagnostic, serologic and  cellular toxicological mechanistic assays (MTT, Annexin V binding, DNA fragmentation and caspase activation assays) and imaging techniques, including flow cytometry, enzyme linked immunosorbant assay (ELISA) and immunohistochemistry(IHC), fluorescence based assays (FISH) for specific protein activation and subcellular localization utilizing confocal microscopy (IP and CHIP interaction assays).

 

ü  In collaboration with Gwen Nichols, MD, a staff Oncologist, I conducted clinical serological and toxicological assays on patient blood samples as part of a clinical study to determine efficacy of chemotherapy regimen in patients with B-cell lymphoma.  Cell cycle changes, apoptotic assays, mitochondrial potential, caspase activation and the expression of key apoptotic mRNAs and key biomarker proteins were measured by flow cytometry (FACS), confocal microscopy cellular imaging, western blot, ELISA and qRT-PCR gene expression analyses.

ü  Experience troubleshooting bioassays and equipment operation;  development and optimization of diagnostic assays in virology and oncology

 

  • Designed and constructed mammalian expression vectors, which expressed micro RNAs, which would interfere and decrease the expression of the anti-apoptotic protein bcl-2.  An anti-sense oligomer, which is in clinical trials, has been shown to decrease the viability of human prostate cancer cells, which decreased bcl-2 protein expression by almost 90%.  Determine DMPK studies with measurements of absorption, metabolism and elimination of labeled anti-sense biological oligomers in cell lines.  These studies were confirmed using siRNAs, which also dramatically decreased bcl-2 expression.  This project suggested that while bcl-2 levels were important in oligomer efficacy, alternative mechanisms, perhaps involving a cellular immune type of response, would also play a key role in anti-sense bcl-2 oligomer action against prostate cancer viability.

 

  • RNA, DNA and protein purification, hybridization, PCR amplification and genetic engineering, cloning, and microarray, Western blot and ELISA immuno-precipitation, FRET, CHIP analysis

 

  • Broad based knowledge and experience in:
    • 10+ years’ experience in cell and tissue culture with primary cells and stable, established cell lines (human, mouse)
    • mammalian, insect and bacterial expression systems with recombinant engineering methodologies (cloning, vector construction/verification, sequence comparison and data mining)
    • experience with potentially highly infective BSL3 environment dealing with active human viruses

 

  • As an advisor to a Ph.D. graduate student I transfected prostate cancer cells with vectors designed to over-express cIAP-1 protein, a key biomarker of chemotherapy resistance.  While I isolated clones that expressed only minimal increases in this protein the cells were resistant to several chemotherapy reagents, however the growth rates of these cells was substantially lower than mock transfected cells, suggesting a possible role for this protein in cell cycle control (yet to be determined).
    • Use of siRNA, antisense oligomers, drug, antibody and peptide therapies
    • Gene expression analysis using flow cytometry (FACS), RT-qPCR, qPCR, sequencing, Northerns, run-off transcription, Westerns, Southerns, FISH, ELIZA, immuno-histochemistry and confocal microscopy
    • Small animals (PI on IACUC proposals), xenograph models of cancer, toxicologic assays of  chemotherapy agents and IHC imaging studies and drug efficacy models in the development of therapeutic agents

 

  • Radiolabelled ligands were designed to bind to a class of endoplasmic reticular (ER) proteins, termed Sigma receptors, a biomarker to highlight cancer cells in PET scanning.  Interestingly, exposure of cancer cells to receptor antagonists has been shown to induce the cell death of prostate cancer cells.  While this cell death has some features of apoptosis, I have some preliminary data that this death maybe a form of apo-necrosis (oncosis or autophagy).  While preliminary, our data suggests that calcium ion transport may be affected. It also has been shown that sigma 1 receptors bind to the inositol-3 phosphate receptor in the ER membrane. This further suggests that bcl-2, known to protect cells against ER calcium release cell death, may also be involved in sigma receptor function and perhaps ligand binding.  

 

Columbia University Cancer Center

  • Setup and organized the micro-array for gene expression analysis core facility of the Institute of Cancer Genetics, Pathology under its director Professor Riccardo Dalla –Favera, PhD.  In this temporary role I supervised the acquisition of equipment and materials, trained the technician who ran the facility and organized the data management; all in cooperation with Affymetrix application scientists.  This facility was designed to determine expression analysis, for pre-clinical and basic research investigations of biomarkers involved in cancer potential and progression

 

 

 

 

 

                                                                                                                                    Sept 1992 - Sept 1995

Department of Urology, Columbia University    New York, NY                                                                                                                        

Associate Research Scientist

 

  • Determined the effect of over-expression and gene copy number analysis of the anti-apoptotic biomarker  protein bcl-2 (bcl-xL) on androgen withdrawal and chemotherapy resistance in human prostate cancer cell lines

Award- American Association for Cancer Research 1994 Travel Award for Meritorious Abstracts.  Raffo, A., Perlman, H., Day, M., Chen, M.W., and Buttyan, R.   New anti-apoptosis gene identified in a prostate cancer cell line.

 

ü  Designed a highly sensitive RT-PCR based assay (chemiluminescence) to detect minimal human prostate cancer cells in the circulation of human prostate cancer patients this assay is used in identifying those patients who would not be a good candidate for the highly evasive radical prostate surgery, because of metastatic circulating prostate cells.  This assay targeted the prostate biomarkers PSA and PSM to highlight these circulating prostate cancer cells.

 

ü Serological and toxicological bio-assays on cancer patient samples

 

Award-American Urological Association. 1995 Best Poster in Disease Staging.  deVries, G., Olsson, C.A., Raffo, A.J., Cam, C., O'Toole, K., Benson, M.C., Buttyan, R. and Katz, A.E.  The Molecular staging of prostate cancer using an RT-PCR assay: a study of 100 radical prostatectomy patients.

 

  • Designed and conducted studies to determine the effect of wild type and mutant p53 over-expression in human prostate cancer cells to hypoxia induced increases in Hif-1 alpha expression and Hif-1 transcriptional activation in the expression of angiogenic proteins controlled by Hif-1.

 

                                                                                                                      Sept. 1994-Sept. 1995

Advisor, Resident Laboratory Research Program, Department of Urology, Columbia University

  • Trained and supervised Urology residences on proper animal handling and cell culture techniques.
  •  Helped residents design and conduct in vivo and in situ experiments in prostate cancer research.

 

 

                                                                                                                                                                                                                                                                                                                  Sept 1987-July 1992 

Department of Plant Pathology, University of California, Riverside, CA                                                                                                 PhD fellow and post-doctoral fellow

 

  • Molecular cloning of DNA probes for RNA viruses. This assays focused on specific biomarkers for several commercially important, in California, viruses included Citrus Tristeza Virus (CTV) and Zucchini (ZMV) and Watermelon Mosaic Viruses (WMV) and conducted diagnostic screening of specific biomarkers for these viruses in commercial plant samples.  This involved Western blot and ELISA analysis of viral coat proteins and PCR or RT-PCR for viral genome detection
  • Generate a sub-genomic defective interfering RNA fragment of Tobacco Mosaic Virus and determine the effect of this fragment on disease symptoms and viral titers in infected Tobacco.  These studies targeted several biomarkers important in viral replication and virus particle assembly.

 

 

                                                                                                                                 August 1983-June 1987 

Dept. of Biochemistry, University of California, Riverside, CA                              

Research Assistant

  • Determine the effects of Vitamin D on the expression of collagen protein and mRNA on new born rat developing bone by runoff transcription assay, Northern and Western blot and ELISA and Dotblots.  Completion of requirements for Master of Science in Biochemistry and Molecular Biology

             

 

                                                                                                                                 Sept. 1979 -July 1983 

Dept. of Orthopedics, Univ. of Southern California at Los Angeles, CA 

Research Associate / Manager 

  • In charge of maintaining the facility’s cell culture and animal facilities 
  • Oversee and train undergraduate, graduate and medical students and residences in proper animal handling and in cell culture techniques.
  • Conducted experiments to determine the effect of cell culturing primary rabbit cartilage on the expression and phenotype of several connective tissue collagen and proteoglycans species being synthesized and identified key biomarker alterations after injury

 

 

         Sept. 1974-July 1979

The Mount Sinai School of Medicine of New York University, New York, NY  

Research Associate/ Manager - Department of Physiology and Biophysics

  • Conducted experiments to determine the role of phosphorylation/dephosphorylation of a small cardiac muscle biomarker endoplasmic reticulum 22KD protein (phospholambin) in the regulation of calcium ion transport out of the endoplasmic reticulum in cardiac muscle in response to cytokine activation.

 

 

 

 

Education

  • University of California at Riverside      PhD,  Plant Pathology,  3.42 GPA  
  • University of California  at Riverside     MS, Biochemistry and Molecular Biology, 3.75 GPA   
  • Hunter College, City Univ. of New York         BA, Biological Sciences, 3.25 GPA   

 

 

 

 

References

 

1)      Matthew Marton, Ph.D., Director of Research Sciences, Clinical Development Laboratories- Genomics, Merck & Co., RY50-1D-134, 126 E Lincoln Ave., Rahway, NJ 07065. matthew_marton@merck.com 732-594-1945.  Supervisor at Merck & Co., CDL-Genomics; designs, develops and validates genomic assays for clinical application of Merck initiated therapeutics.

2)      Paul W. Brandt-Rauf. MD, Ph.D., Dean, School of Public Health, University of Illinois, 1175 SPHPI, MC 923, Chicago, IL 60612. pwb1@uic.edu 312-996-5939 Prior supervisor and Principle Investigator for projects involving p53 activation by a p53 derived peptide, including binding of peptide to p53 and structural characterization of peptide bound/unbound to p53.  He is also the Principle Investigator on studies investigating the molecular and structural effects of polymorphisms of a key DNA repair scaffolding protein XRCC1.

3)      Ramon V. Rosal, PhD., CT Coordinator/Chemical Response Director, Environmental Sciences and Toxicology, Public Health Labs, The City of New York, Dept. of Health & Mental Hygiene, 455 First Ave, rm 1210, New York, NY 10016.  rrosal@health.nyc.gov, 212-447-1149.  Collaborator on the project investigating the p53 derived C-terminal peptide: p53 activation, peptide/mutant p53 binding studies and structural characteristics of peptide bound/un-bound to p53.

4)      Cy A. Stein, MD, Ph.D., Professor, Montefiore Medical Center, Dept. of Oncology, 111 E. 210 St., The Bronx, NY 10467, cstein@montefiore.org, 718-920-8980.  Prior supervisor and Principle Investigator on projects involving anti-sense RNA, oligomers and siRNA against bcl-2 family pro-life proteins and IAP overexpression.

5)      Ralph Buttyan, Ph.D., Professor; Division of Urology, The Ordway Research Institute; Cancer Center; CMS 4th Floor; 150 New Scotland Ave, Albany, New York, 12208, buttyanr@acp.edu . 518.641.6980. Prior supervisor and Principle Investigator on projects dealing with the bcl-2 family of proteins and the effects of castration and hypoxia on prostate cancer cells and design of PCR based detection studies of circulating prostate cancer cells in the blood of prostate cancer patients.

6)      Lloyd A. Greene, Ph.D., Professor, Dept. of Pathology, Columbia University, 630 W 168 St., New York, NY 10032 lag3@columbia.edu 212-305-6369 Chief Principle Investigator overseeing Apoptosis journal club

7)      Benya, Paul PhD, Adjunct Associate Professor, Department of Orthopaedic Surgery, UCLA / Orthopaedic Hospital, David Geffen School of Medicine, Orthopaedic Hospital Research Center, 615 Charles E. Young Drive South, Los Angeles, CA 90095 310-983-1035. Prior Supervisor and Principle Investigator on projects dealing with the de-differentiation of chondrocytes after wound healing and the characterization of proteins and cell surface biomarkers that resulted.

8)      William Dawson, Ph.D., Professor, Department of Plant Pathology, University of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850, fax: 1-863-956-4631, wodtmv@C.REC.iFAS.ufl.edu  PhD mentor and principle investigator on plant viral molecular studies and diagnostic assay development.

 

 

 

Publications:

 

1. Kirchberger, M.A., and Raffo, A.  Phosphoprotein phosphatase-catalyzed dephosphorylation of the 22,000-dalton phosphoprotein of cardiac sarcoplasmic reticulum. Recent Adv Stud Cardiac Struct Metab. 11: pp.285-91, 1976.

2. Kirchberger, M.A. and Raffo, A.J.  Decrease in calcium transport associated phosphoprotein phosphatase-catalyzed dephosphorylation of cardiac sarcoplasmic  reticulum.   J. of Cyclic Nucleotide Research  3: pp. 45-53, 1977.

3.  Benya, P.D., Jaffe, S. and Raffo, A.J.  The capacity of chondrocytes to respond to serum is enhanced by organ culture in the absence of serum, stimulated by serum and modified by ascorbate.  Archives of Biochemistry and Biophysics  232:  pp. 323-336, 1984.

4.  Dawson, W.O., Lewandowski, D.J., Hilf, M.E., Bubrick, P., Raffo, A.J., Shaw, J.J., Grantham, G.L. and Desjardins, P.R.  A tobacco mosaic virus-hybrid expresses and loses an added gene.  Virology  172: pp. 285-292, 1989.

5.  Raffo, A.J., Khan, I.A., Lippert, L.F., Hall, M.O. and Jones, G.E. A screening procedure for ZYMV resistance in muskmelons.  Cucurbit Genetics Cooperative Report  12: pp. 46-49, 1989.

6.  Raffo, A.J. and Dawson, W.O.  Construciton of tobacco mosaic virus subgenomic replicons that are replicated and spread systemically in tobacco plants.  Virology 184: pp 277-289, 1991.

7.  Zhang, X., Colombel, M., Raffo, A., Buttyan, R.  Enhanced expression of p53 mRNA and protein in the regressing rat ventral prostate gland. Biochem. Biohys. Res. Commun.  198: pp. 1189-1194, 1994.

8.  Katz, A.E., Olsson, C.A., Raffo, A.J., Cama, C., Perlman, H., Seaman, E., McMahan, D., Benson, M.C. and Buttyan, R.  Molecular staging of prostate cancer with the use of an enhanced reverse transcriptase-PCR assay. Urology  43: pp. 765-775, 1994.

9.  Raffo, A., Perlman, H., Day, M., Chen, M.W., and Buttyan, R.  New anti-apoptosis gene identified in a prostate cancer cell line.  Proc. Amer. Assn. Cancer Res. 35: p. 23, 1994.

10.  Cama, C., Olsson, C.A., Raffo, A.J., Perlman, H., Buttyan, R., O'Toole, K.M., McMahon, D., Benson, M.C. and Katz, A.E.  Molecular staging of prostate cancer II: A comparison of the application of an enhanced reverse transcriptase polymerase chain reaction assay for prostate specific antigen versus prostate specific membrane antigen. The Journal of Urology  153: pp 1373-1378, 1995.

11.  Katz, A.E., de Vries, G.M., Begg, M.D., Raffo, A.J., Cama, C., O'Toole, K., Buttyan, R., Benson, M.C., Olsson, C.A.  Enhanced reverse transcriptase polymerase chain reaction for prostate specific antigen as an indicator of true pathological stage in patients with prostate cancer.   Cancer 75: pp. 1642, 1995.

12.  Sensibar, J.A., Sutkowski, D.M., Raffo, A.J., Buttyan, R., Griswold, M.D., Sylvester, S.R., Kozlowski, J.M. and Lee, C.  Prevention of cell death induced by tumor necrosis factor alpha in LNCaP cells by overexpression of sulphated glycoprotein-2 (custerin).  Cancer Research 55: pp. 2431-2437, 1995.

13.  Raffo, A.J., Perlamn, H., Chen, M.W., Day, M.L., Streitman, J.S. and Buttyan, R.  Overexpression of bcl-2 protects prostate cancer cells from apoptosis in vitro and confers resistance to androgen-depletion in vivo.  Cancer Research 55: pp. 4438-4445, 1995.

14.  de Vries, G.M., Olsson, C.A., Raffo, A.J., Cama, C., O'Toole, K., Benson, M.C., Buttyan, R. and Katz, A.E.  The molecular staging of prostate cancer using an RT-PCR assay: a study of 100 radical prostatectomy patients. J. Urology  153: p 294A, 1995.

15.  Cama, C.,  Olsson, CA., Raffo, A.J., Perlmon, H., Buttyan, R., O'Toole, K., McMahon, D., Benson, M.C. and Katz, E.  Molecular staging of prostate cancer II.  A comparison of the application of an enhanced reverse transcriptase polymerase chain rector assay for prostate specific antigen versus prostate specific membrane antigen.  J. of Urology 153: pp 1373-1378, 1995.

16.  Olsson, C. A., de Vries, G.M. , Raffo, A.J., Buttyan, R., Benson, M.C., Cama, C., O'Toole, K. and Katz, A.E.  The use of RT-PCR for prostate  specific antigen assay to predict potential surgical failures before radical prostatectomy: molecular staging of prostate cancer. British J. of Urology  77: pp. 411-417, 1996.

17.  Olsson, C. A., de Vries, G.M. , Raffo, A.J., Benson, M.C., O'Toole, K.,  Cao, Y., Buttyan, R. and Katz, A.E. Pre-operative RT-PCR for PSA can predict treatment failure following radical prostatectomy.  J. of Urology  155: pp. 1557-1562, 1996.

18.  Klein,  L.T., Miller, M.I., Buttyan, R., Raffo, A.J., Burchard, M., DeVris, G., Cao, Y.C., Olsson, C. and Shabsigh, R.  Apoptosis in the rat penis after penile denervation.  J. Urology 158: pp 626-630, 1997.

19. Kim, A.L., Raffo, A.J., Brand-Rauf, P.W., Pincus, M.R., Monaco, R., Abarzua, P. and Fine, R.L. Conformational and molecular basis for induction of apoptosis by a p53 C-terminal peptide in human cancer cells.  The Journal of Biological Chemistry 274(4), pp. 34924-34931, 1999.

20. Raffo, A.J., Kim, A.L. and Fine, R.L.  The formation of a nuclear p53/Bax complex is associated with chemotherapy induced apoptosis. Oncogene  Dec 14;19(54):6216-6228, 2000

21. Lebedeva, I., Raffo, A.J., Rando, R., Ojwang, J., Cossum, P. and Stein, C.A. Chemosensitization of bladder carcinoma cells by Bcl-xL antisense oligonucleotides. J Urol. Aug;166(2):461-469, 2001

22. Kanovsky, M., Raffo, A., Drew, L., Rosal, R., Do, T., Friedman, F.K., Rubinstein, P., Visser, J., Brandt-Rauf, P.W., Fine, R., Michl, J. and Pincus, M.R.  Peptides from the amino terminal mdm2 binding domain of p53 designed from conformational analysis are selectively cytotoxic to transformed cells.  Proc. Natl. Acad. Sci. USA Oct 23; 98(22):pp.12438-12443, 2001.

23. Vilenchik, M., Raffo, A.J., Shames, D. and Stein, C.A. Antisense RNA downregulation of bcl-xL expression in prostate cancer cells leads to diminished rates of cellular proliferation and resistance to cytotoxic chemotherapeutic agents. Cancer Research April 01:62:pp. 2175-2183, 2002

24. Li, Y., Raffo, A.J., Drew, L., Mao, Y., Tran, A., Petrylak, D.P., and Fine, R.L. Fas-mediated apoptosis is dependent on wild-type p53 status in human cancer cells expressing a temperature-sensitive p53 mutant alanine-143. Cancer Res  Apr 1;63(7):1527-33, 2003

25. Do, T.N., Rosal, R.V., Drew, L., Raffo, A.J., Michl, J., Pincus, M.R., Friedman, F.K., Petrylak, D.P., Cassai, N., Szmulewicz, J., Sidhu, G., Fine, R.L. and Brandt-Rauf, P.W.  Preferential induction of necrosis in human breast cancer cells by a p53 peptide derived from the MDM2 binding site. Oncogene Mar 13;22(10):1431-44, 2003

26. Raffo, A.J., Lai, J.C., Stein, C.A., Miller, P., Scaringe, S., Khvornova, A. and Benimetsky, L.  Antisense RNA downregulation of bcl-2 expression in DU145 prostate cancer cells does not diminish the cytostatic effects of G3139 (Oblimersen). Clinical Cancer Research, May 1, 10(9):3195-3206, 2004

27. de Souza, F., Simpson, N., Raffo, A.J., Saxena, C., Maffei, A., Hardy, M., Goland, R., Libel, R., Mann, J., Van Heertum, R., and Harris, P.E. Longitudinal noninvasive P.E.T. based beta cell mass estimates in a spontaneous diabetes rat model. J. Clinical Investigation. Jun; 116(6):1506-1513, 2006

28. Simpson, N., de Souza, F., Witkowski, P., Maffei, A., Raffo, A.J., Herron, A., Kilbourn, M., Herold, K., Liu, E., Hardy, M., Van Heertum, R. and Harris, P.E. Visualizing pancreatic beta-cell mass with [(11)C]DTBZ. Nucl Med Biol. Oct; 33(7):855-864, 2006

29. Hultman K.L., Raffo, A.J., Grzenda, A.L., Harris, P.E., Brown, T.R. and O’Brien, S. Magnetic resonance imaging of major histocompatibility class II expression in the renal medulla using immunotargeted superparamagnetic iron oxide nanoparticles. Amer. Chem. Society-Nanotechnology.  Mar; 2(3):477-484, 2008

30. Xie, Y., Raffo, A., Ichise, M., Deng, S., Harris, P.E., and Landry, D.W.  Novel hypoglycemic dihydropyridones serendipitously discovered from O- versus C-alkylation in the synthesis of VMAT2 antagonists.  Bioorg Med Chem Lett.  Sep 18(18):5111-4, 2008

31. Raffo, A., Hancock, K., Polito, T., Xie, Y., Andan, G., Witkowski, P., Hardy, M., Barba, P., Ferrara, C., Maffei, A., Freeby, M., Goland, R., Leibel, R.L., Sweet, I.R., and Harris, P.E.

Role of vesicular monoamine transporter type 2 in rodent insulin secretion and glucose metabolism revealed by its specific antagonist tetrabenazine.  J Endocrinol. July 198(1):41-9. 2008

32. Raffo, A.  :Genotyping of human Beta-Catenin exon 3, by Sanger Sequencing, as part of the MK0000-215 biomarker identification project.: Core Needle Biopsy Equivalents (CNBE) using formalin-fixed, paraffin-embedded biopsy tissue sections (FFPE) Internal Merck Validation Report, VAL834, 2013

33. Raffo, A and Chen, Lei.  Design and validation of two real-time PCR based, “Taqman” assays designed to identify two distinct single nucleotide polymorphisms (SNPs) within the Homo sapiens lipoprotein (a), LPA gene using DNA isolated from predominately Paxgene treated human blood samples.  Real-time PCR based SNP genotyping assay with Sanger Sequencing validation of assay positive controls.  Internal Merck Validation Report,  VAL840 2013.

 

 

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