Paul A. Azzinaro is a master scientist seeking employment who has extensive experience in molecular biology, cell biology, and proteomics. He received his Master's degree in cell and molecular biology from the University of Rhode Island, where he investigated the Fanconi anemia DNA repair pathway. His laboratory experience includes work at Plum Island Animal Disease Center developing vaccines and using yeast-2-hybrid and immunofluorescence techniques, as well as previous positions conducting assays and as a teaching assistant.
Microbiome Isolation and DNA Enrichment Protocol: Pathogen Detection Webinar ...QIAGEN
This slidedeck presents an easy-to-use workflow that allows selective isolation of microbial DNA from samples that are intrinsically rich in host DNA. This protocol includes steps for efficient depletion of host DNA while providing optimized conditions specific for bacterial lysis. This workflow is also specific for the identification of live bacteria, avoiding false results due to nucleic acids from dead bacteria. Enriched microbial DNA can be directly used in other molecular methods such as whole genome sequencing, qPCR and microarray assays.
Clinical Metagenomics for Rapid Detection of Enteric Pathogens and Characteri...QIAGEN
High-throughput sequencing, combined with high-resolution metagenomic analysis, provides a powerful diagnostic tool for clinical management of enteric disease. Forty-five patient samples of known and unknown disease etiology and 20 samples from health individuals were subjected to next-generation sequencing. Subsequent metagenomic analysis identified all microorganisms (bacteria, viruses, fungi and parasites) in the samples, including the expected pathogens in the samples of known etiology. Multiple pathogens were detected in the individual samples, providing evidence for polymicrobial infection. Patients were clearly differentiated from healthy individuals based on microorganism abundance and diversity. The speed, accuracy and actionable features of CosmosID bioinformatics and curated GenBook® databases, implemented in the QIAGEN Microbial Genomics Pro Suite, and the functional analysis, leveraging the QIAGEN functional metagenomics workflow, provide a powerful tool contributing to the revolution in clinical diagnostics, prophylactics and therapeutics that is now in progress globally.
This presents a number of case studies on the application on high-throughput sequencing (HTS), next generation sequencing (NGS), to biological problems ranging from human genome sequencing, identification of disease mutations, metagenomics, virus discovery, epidemic, transmission chains and viral populations. Presented at the University of Glasgow on Friday 26th June 2015.
Microbiome Isolation and DNA Enrichment Protocol: Pathogen Detection Webinar ...QIAGEN
This slidedeck presents an easy-to-use workflow that allows selective isolation of microbial DNA from samples that are intrinsically rich in host DNA. This protocol includes steps for efficient depletion of host DNA while providing optimized conditions specific for bacterial lysis. This workflow is also specific for the identification of live bacteria, avoiding false results due to nucleic acids from dead bacteria. Enriched microbial DNA can be directly used in other molecular methods such as whole genome sequencing, qPCR and microarray assays.
Clinical Metagenomics for Rapid Detection of Enteric Pathogens and Characteri...QIAGEN
High-throughput sequencing, combined with high-resolution metagenomic analysis, provides a powerful diagnostic tool for clinical management of enteric disease. Forty-five patient samples of known and unknown disease etiology and 20 samples from health individuals were subjected to next-generation sequencing. Subsequent metagenomic analysis identified all microorganisms (bacteria, viruses, fungi and parasites) in the samples, including the expected pathogens in the samples of known etiology. Multiple pathogens were detected in the individual samples, providing evidence for polymicrobial infection. Patients were clearly differentiated from healthy individuals based on microorganism abundance and diversity. The speed, accuracy and actionable features of CosmosID bioinformatics and curated GenBook® databases, implemented in the QIAGEN Microbial Genomics Pro Suite, and the functional analysis, leveraging the QIAGEN functional metagenomics workflow, provide a powerful tool contributing to the revolution in clinical diagnostics, prophylactics and therapeutics that is now in progress globally.
This presents a number of case studies on the application on high-throughput sequencing (HTS), next generation sequencing (NGS), to biological problems ranging from human genome sequencing, identification of disease mutations, metagenomics, virus discovery, epidemic, transmission chains and viral populations. Presented at the University of Glasgow on Friday 26th June 2015.
1. Paul A. Azzinaro Curriculum Vitae
Master Scientist for Hire
Address: 125 Tower St. Westerly, RI, 02891 E-mail: paul_azzinaro@my.uri.edu
Phone: 401-932-6683
Education
2011- 2014 Master of Science, Biological and Environmental Sciences, Cell and Molecular Biology,
Date of graduation: December 2014
University of Rhode Island, Kingston, RI
Advisor: Dr. Niall Howlett
2008- 2011 Bachelor of Science, Biology, May 2011
University of Rhode Island, Kingston, RI
Laboratory Experience
Mar 2016-Present Collaborating Scientist
Mar2016-Present PlumIslandAnimalDiseaseCenter, Orient Point, NY. I have used my background in molecular
biology to help develop vaccines against agricultural diseases with large economic consequences
impacting the food security of the country. This work is conducted in a BSL 3 environment. My
work had focused on using Yeast-2-Hybrid to screen for interactions between the virus and host
proteins. Using Immunofluorescence interactions are than confirmed within actively infected cells.
This work supports the construction of an attenuated virus usable as a vaccine strain in a disease
which currently lacks any means of treatment.
June 2015-Mar 2016 Laboratory Technician
June2015-Mar2016 GSM Metals, Cranston, RI. I worked in a lab conducting assays on alloys of precious metals used
in both jewelry and industry. This lab had a large focus on accurate and quick data reporting to
ensure all materials were within the correct specifications.
Jan 2015-May 2015 Proteomics consultant
Jan2015-May2015 University of Rhode Island, Kingston, RI. I have been contacted for consulting opportunities to
advise on experimental design, and to teach and conduct proteomic analysis. This opportunity has
focused on the use of mass spectrometry, sample preparation and analysis of data using the
ProteoIQ platform.
Aug 2011-Dec 2014 Graduate Assistant/Research Assistant, URI
Jan2012-Dec2014 Dr. Niall Howlett’s Lab, Kingston, RI. The goal of the lab is to investigate human gene and protein
function in Fanconi anemia, a genetic disease resulting in DNA damage repair defects. My
Master’s thesis was titled “Identifying novel FANCD2 interacting proteins via immunoprecipitation
Paul A. Azzinaro Curriculum Vitae 1
2. and mass spectrometry.” This work involved designing and optimizing protocols for using
immunoprecipitations to enrich for immune complexes and subsequently using proteomics and
mass spectrometry to identify the immune complex components. I was also heavily involved with
identifying and characterizing new domains in our protein of interest, FANCD2, in an attempt to
better understand the regulation and function of the protein. The lab predominantly used cell based
systems for experimentation requiring all members to have strong experience with mammalian
tissue culture. Tissure culture was important for working with patient derived lines, performing
transient transfections, and generating stable cell lines using molecular cloning and site directed
mutagenesis with a Lenti viral packaging system. Functional characterizations were most
commonly performed using SDS-PAGE and Western blotting to look at protein expression and post
translational modification. Other commonly used assays were cell survival assays and
immunofluorescence. I was involved in several side projects which gave me experience with
techniques such as molecular modeling, flow cytometry, 2D gels, and computer programming.
Nov2011-Dec2011 Dr. Bethany Jenkins’ Lab (5 weeks). Kingston, RI. During my brief rotation in the Jenkins lab,
which focuses on microbial ecology and the role of diatoms in nutrient cycling, I spent most of my
time using q-RT-PCR to investigate differential expression in nutrient limited diatoms.
Oct2011-Nov2011 Dr. Paul Cohen’s Lab (5 weeks). Kingston, RI. The focus of the Cohen lab is to study the
probiotic effects of specific strains of E. coli against pathogenic strains using the streptomycin
treated mouse model. During this rotation I performed mouse intestinal colonizations and used cell
counting methods to determine the ability for different strains of E. coli to colonize the mouse
gastro intestinal tract.
May 2010- Aug 2011 Laboratory Assistant
May2010-Aug2011Rhode Island Analytical Laboratories, Warwick, RI. This laboratory is an analytical chemistry
laboratory focused on environmental testing. My responsibilities while employed at this company
were managing inventory as well as performing basic analytical tests such as total solids, total
suspended solids, and oil and grease testing.
Sep2010-May2011 Dr. Steven Irvine’s Laboratory, Kingston, RI. The Irvine laboratory studies sea squirts a marine
invertebrate chordate. During my time in this lab I used PCR and molecular cloning to work on
identifying microbial endosymbionts unique to invasive species of tunicates. I had experience using
electroporation on eukaryotic cells and lots of microscopy experience.
Teaching Experience
Aug 2011-Dec 2014 Teaching assistant, University of Rhode Island
Lab coordinator/instructor:
Introductory Biochemistry Laboratory (BCH 312), Aug 2013-May 2014, Kingston, RI.
Assisted in designing labs and class setup, lectured on class material. The purpose of this
lab was to give students experience with modern techniques often used in molecular
biology labs focusing on cloning a mutant plasmid, expressing the protein in E. coli,
purifying that protein, performing pharmacokinetics on the purified protein, and using
molecular modeling to visualize the reaction.
Lab instructor:
Introduction to Microbiology (MIC 201), Aug 2012-Dec 2014, Kingston, RI. Lectured on
class material, writing and grading of laboratory quizzes and exams. This class served to
Paul A. Azzinaro Curriculum Vitae 2
3. familiarize students with microbes, aseptic techniques and methods to identify microbial
species.
Human Anatomy (BIO 121), Aug 2011-May 2012, Kingston, RI. wrote weekly quizzes,
graded quizzes and practicals, assisted in test proctoring. This class was designed to
familiarize students interested in health care with the human body.
Laboratory Skills
Cell culture
• Mammalian cell culture
• Transient transfections
• Cell survival assays
• Lenti viral packaging
• Microscopy
• Immunofluorescence microscopy
• Flow cytometry
Molecular biology
• Molecular cloning
• PCR
• Site directed mutagenesis
• UV-Vis spectrometry
• Yeast-2-Hybrid
• Bacterial and yeast transformations
Cell Biology
• Western Blotting
• Immunoprecipitations
• Mass spectrometry
• Proteomics
• Molecular modeling
Other
• Microsoft office
• Microsoft excel
• Microsoft PowerPoint
• R studio
• Ordering
• Making buffers and reagents
• Data recording and analysis
• Designing and optimizing protocols and experiments
• Fire Assay
• Titration
• ICP
• BSL-2
• BSL-3
Paul A. Azzinaro Curriculum Vitae 3
4. Certifications
FAES “Proteomics: Principles and Methods” workshop (40 hour)
Bloodborne Pathogens and Biosaftey Training
Lab Safety and Hazardous Waste Training
NAUI advanced diver certification
Outreach
2013-2014 Mentor: Trained an undergraduate in techniques used in our laboratory as part as an
Honors program requirement for the student.
References
Graduate advisor
Dr. Niall H. Howlett
Niall G. Howlett, Ph.D., Associate Professor Department of Cell and Molecular Biology University of Rhode
Island 379 CBLS, 120 Flagg Road Kingston, RI 02881
Undergraduate advisor
Dr. Steven Irvine
Steven Q. Irvine, Ph.D., Associate Professor Department of Biological sciences University of Rhode Island 281
CBLS, 120 Flagg Road Kingston, RI 02881
Publications
O'Donnell V, Risatti GR, Holinka LG, Krug P, Carlson J, Velazquez-Salinas L, Azzinaro PA, Gladue DP,
Borca MV. Simultaneous deletion of the 9GL and UK genes from the African swine fever virus Georgia
2007 isolate offers increased safety and protection against homologous challenge. J Virol. 2016 Oct 26. pii:
JVI.01760-16.
Borca MV, O'Donnell V, Holinka LG, Rai DK, Sanford B, Alfano M, Carlson J, Azzinaro PA, Alonso C,
Gladue DP. The Ep152R ORF of African swine fever virus strain Georgia encodes for an essential gene
that interacts with host protein BAG6. Virus Res. 2016 Sep 2;223:181-9. doi:
10.1016/j.virusres.2016.07.013. Epub 2016 Aug 3.
Stanley EC, Azzinaro PA, Vierra DA, Howlett NG, Irvine SQ. The Simple Chordate Ciona intestinalis Has a
Reduced Complement of Genes Associated with Fanconi Anemia. Evol Bioinform Online. 2016 Jun
6;12:133-48. doi: 10.4137/EBO.S37920. eCollection 2016.
Paul A. Azzinaro Curriculum Vitae 4
5. Azzinaro PA, Mauro M, Howlett NG. Identification Of Novel Fancd2 Interacting Proteins Via
Immunoprecipitation And Mass Spectrometry. Proquest. 2014 Dec 03 Available 2016 Dec 03.
Boisvert RA, Rego MA, Azzinaro PA, Mauro M, Howlett NG. Coordinate nuclear targeting of the FANCD2
and FANCI proteins via a FANCD2 nuclear localization signal. PLoS One. 2013 Nov 21;8(11):e81387.
Grants, Awards and honors
2012, 2014 URI Student Research and Travel Fund
2011 Alpha Award for outstanding achievement in research in the Biological
Sciences Department at URI
2008 Boy Scouts of America, Eagle Scout
Poster Presentations
2014 Azzinaro, P.A., Howlett, N.G. Identification of Novel Chromatin-Associated FANCD2 Interacting
Proteins. 26th
Annual Fanconi Anemia Research Foundation Scientific Symposium. September
18-21, Bethesda, MD.
2014 Paquin, K.L, Vierra, D.A., Azzinaro, P.A., Rego, M.A., Howlett, N.G. Characterization of a Putative
Histone Binding Domain in FANCD2. 26th
Annual Fanconi Anemia Research Foundation Scientific
Symposium. September 18-21, Bethesda, MD.
2014 Paquin K.L., Azzinaro, P.A., Howlett, N.G. Chromatin Targeting of the Fanconi Anemia D2 Protein.
Biological Methylation: Regulation of Chromatin, Epigenetics, and Disease. July 6-11, Nassau,
Bahamas.
2014 McClanaghan, J.D., Paquin, K.L., Azzinaro, P.A., Howlett, N.G. Defining the sites of interaction of
the FANCD2, FANCL, and FANCE proteins. Honors Annual Conference. May 1, Kingston, RI.
2013 Paquin, K.L., Azzinaro, P.A., Howlett, N.G. Identification and Characterizaion of a Putative Histone
Binding Domain in FANCD2. 7th
Annual NESP-BioNES Meeting. December 6, 2013, Bristol, RI.
2013 Lima, K., Azzinaro, P.A., Howlett, N.G. Characterizaion of a Putative Chromodomain in the
Fanconi Anemia D2 Protein. 7th
Annual NESP-BioNES Meeting. December 6, 2013, Bristol, RI.
2013 Neira, K.L., Mauro, M., Azzinaro, P.A., Howlett, N.G. Chromatin Dynamics and the Fanconi
Anemia DNA Damage Response Pathway. Experimental Biology. April 19-24, Boston, MA.
2013 Howlett, N.G., Rego, M.A., Boisvert, R.A., Neira, K.L., Azzinaro, P.A. Novel Insight into the
Fanconi Anemia DNA Repair Pathway by Domain Structure-Function Studies of FANCD2.
Keystone Symposia. March 3-8, 2013, Canada
Paul A. Azzinaro Curriculum Vitae 5
6. Azzinaro PA, Mauro M, Howlett NG. Identification Of Novel Fancd2 Interacting Proteins Via
Immunoprecipitation And Mass Spectrometry. Proquest. 2014 Dec 03 Available 2016 Dec 03.
Boisvert RA, Rego MA, Azzinaro PA, Mauro M, Howlett NG. Coordinate nuclear targeting of the FANCD2
and FANCI proteins via a FANCD2 nuclear localization signal. PLoS One. 2013 Nov 21;8(11):e81387.
Grants, Awards and honors
2012, 2014 URI Student Research and Travel Fund
2011 Alpha Award for outstanding achievement in research in the Biological
Sciences Department at URI
2008 Boy Scouts of America, Eagle Scout
Poster Presentations
2014 Azzinaro, P.A., Howlett, N.G. Identification of Novel Chromatin-Associated FANCD2 Interacting
Proteins. 26th
Annual Fanconi Anemia Research Foundation Scientific Symposium. September
18-21, Bethesda, MD.
2014 Paquin, K.L, Vierra, D.A., Azzinaro, P.A., Rego, M.A., Howlett, N.G. Characterization of a Putative
Histone Binding Domain in FANCD2. 26th
Annual Fanconi Anemia Research Foundation Scientific
Symposium. September 18-21, Bethesda, MD.
2014 Paquin K.L., Azzinaro, P.A., Howlett, N.G. Chromatin Targeting of the Fanconi Anemia D2 Protein.
Biological Methylation: Regulation of Chromatin, Epigenetics, and Disease. July 6-11, Nassau,
Bahamas.
2014 McClanaghan, J.D., Paquin, K.L., Azzinaro, P.A., Howlett, N.G. Defining the sites of interaction of
the FANCD2, FANCL, and FANCE proteins. Honors Annual Conference. May 1, Kingston, RI.
2013 Paquin, K.L., Azzinaro, P.A., Howlett, N.G. Identification and Characterizaion of a Putative Histone
Binding Domain in FANCD2. 7th
Annual NESP-BioNES Meeting. December 6, 2013, Bristol, RI.
2013 Lima, K., Azzinaro, P.A., Howlett, N.G. Characterizaion of a Putative Chromodomain in the
Fanconi Anemia D2 Protein. 7th
Annual NESP-BioNES Meeting. December 6, 2013, Bristol, RI.
2013 Neira, K.L., Mauro, M., Azzinaro, P.A., Howlett, N.G. Chromatin Dynamics and the Fanconi
Anemia DNA Damage Response Pathway. Experimental Biology. April 19-24, Boston, MA.
2013 Howlett, N.G., Rego, M.A., Boisvert, R.A., Neira, K.L., Azzinaro, P.A. Novel Insight into the
Fanconi Anemia DNA Repair Pathway by Domain Structure-Function Studies of FANCD2.
Keystone Symposia. March 3-8, 2013, Canada
Paul A. Azzinaro Curriculum Vitae 5