Speaker; Dr. John Cipollo, FDA/CBER

Topic: Understanding carbohydrate dependent host-pathogen interactions in the surrogate host Caenorhabditis elegans: A mass spectrometry based approach

Topic: Understanding carbohydrate dependent host-pathogen interactions in the surrogate host Caenorhabditis elegans: A mass spectrometry based approach

Date: Monday, May 14, 2012

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract: Caenorhabditis elegans is infected by over 40 microbial pathogens most of which are human pathogens or their close relatives. Genetic and biochemical evidence suggests that some of these pathogenic interactions are carbohydrate dependent. Here we use a mass spectrometry based approach to investigate the nature of this carbohydrate dependence through comparison of wild type infection susceptible nematode strains and carbohydrate deficient infection resistant strains. The production and use of chemical probes including saccharide loaded latex beads and 2 types of affinity glycoproteomics reagents will be discussed. Also to be discussed will be our efforts towards construction of C. elegans native glycan arrays.

Topic: Protein Processor: Probabilistically Using Mass Accuracy and the MS Spectrum

Date: Monday, April 16, 2012

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract: Current approaches to protein identification rely heavily on database matching of fragmentation spectra, while ignoring (in a scoring sense) the mass accuracy of the precursor ions.  We have developed a method based originally upon MALDI TOF-TOF instrumentation that uses targeted peptide mass fingerprinting results to confirm MS/MS database search identifications.  The method uses 1st-order spectral data that have heretofore been ignored by most search engines, as only a subset of the peptides found in a first order mass spectrum are usually fragmented.  In this method the distribution of mass errors of peptide matches in the first order spectrum is used to develop a probability model that is independent from the MS/MS database search identifications.  Peptide mass matches can come from both precursor ions that have been fragmented as well as those that are tentatively identified by accurate mass alone.  This additional confirmation enables us to assign protein identifications to MS/MS based scores that are otherwise considered to be only of moderate quality.

Topic: Role of Mass Spectrometry in Unveiling Neurotrophic Mechanisms of Docosahexaenoic Acid

Date: Monday, March 19, 2012

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract: Docosahexaenoic acid (DHA, 22:6n-3) is particularly enriched in neuronal tissues mainly as membrane phospholipids.  Maintenance of a high DHA concentration in brain is essential for proper neurodevelopment and function, indicating an important neurotrophic role played by this fatty acid.  Mass spectrometry has been an essential tool for studying molecular and signaling mechanisms underlying DHA-mediated neurotrophic effects.  DHA promotes neuronal survival primarily due to its unique property to increase phosphatidylserine (PS), the major anionic phospholipid in neuronal membranes.  Membrane translocation and activation of key kinases such as Raf, PKC and Akt is PS-dependent, providing a target for DHA-mediated neuroprotection.  DHA is also metabolized to a potent synaptogenic compound synaptamide (N-docosahexaenoylethanoamide), promoting neurite growth, synaptogenesis and synaptic protein expression and improving glutamatergic synaptic function in developing neurons.  Role of mass spectrometry in probing the membrane phospholipid profile, lipid metabolism, protein conformational changes due to membrane-protein interaction, and protein expression will be discussed.

Topic: Top-Down Mass Spectrometry for the Rapid Identification of Strain Specific Bacterial Proteins

Date: Monday, February 13, 2012

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract: Faster and more effective methods for bacterial assessment are becoming increasingly relevant.  Intact protein expression profiling by LCMS is a powerful tool for bacterial strain differentiation.  However, transfer to field usable assays or monitoring of drift in these species requires identification of marker proteins. The availability of predicted proteins from a rapidly expanding database of bacterial genomes affords the possibility of developing tools in which partial protein sequence information obtained from top-down mass spectrometry can rapidly identify strain specific proteins. Combined with the associated molecular weight information, the approach can identify variations in protein sequence that are related to bacterial strain differences. In addition, comparative proteogenomics of closely related bacteria by top-down MSMS allows us to go beyond identification of possible cross-serovar orthologs. Access to intact protein masses, HPLC retention times, and site specific MSMS data provides insight into specific mutations, PTMs, start site errors and signal peptide cleavages that are often poorly annotated in sequence databases. However, intact protein analysis does present its own set of limitations. Data will also be presented illustrating why the current state of intact protein analysis generally yields significantly fewer protein masses than are actually present in the bacteria.

Topic: Rapid and Enhanced Proteolytic Digestion using Electric-Field-oriented Enzyme Reactor

Date: Monday, January 23, 2012

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract: We have created a novel enzyme reactor using electric field-mediated orientation and immobilization of proteolytic enzymes (trypsin/chymotrypsin) on biocompatible PVDF membranes in a continuous flow-through chamber. Using less than 5 minutes, this reactor in various enzyme combinations can produce enhanced rapid digestion for standardized prototypic proteins, hydrophilic proteins and hydrophobic transmembrane proteins when compared to in-solution techniques. With improved digestive efficiency, our reactor improved the overall functional analysis of lipid raft proteomes by identifying more closely functionally linked proteins and elucidated a richer set of biological processes and pathways linked to the proteins than traditional in-solution methods.

Topic: Molecular Imaging of Brain Lipids in Health and Disease

Date: Monday, December 19, 2011

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract:

Lipids are a major component of brain tissue, accounting for almost half of the brain dry weight. They are major building blocks of biological membranes and also play an important role in cell signaling and cell death. Altered levels of lipids in brain tissue are associated with diseases such as Alzheimer’s, Tay-Sachs, and Niemann-Pick disease. MALDI-MS methods have been developed for the in situ analysis of biomolecules, typically peptides and proteins, from tissue. We used MALDI-MS for direct analysis and imaging of lipids in tissue. We have advanced the field of imaging as our work not only maps the distribution of lipids in various areas of the brain it also gives detailed structural information with histological accuracy.  After mapping the content and distribution of lipids in the normal brain, we have demonstrated the changes that occur over time in the lipid make up of the brain in blast induced and mechanical brain injury in animal models.

Speaker: Dr.  Robert Hettich, Senior Staff Scientist, Oak Ridge National Lab

Topic: Development of a MS-based proteogenomic approach for characterization of the functions and metabolic activities of the human gut microbiome

Date: Monday, November 14, 2011

Time: 7:30 pm

Location: Johns Hopkins University School of Medicine (Host: Bob Cotter); dinner at Bertha’s in Fells Point at 5:30 pm

Dinner at Berthas, 734 S. Broadway, Fells Point, MD

Dinner is served family style and includes: Steamed mussels with dipping sauces, button mushrooms with cheddar, mixed salad, greens, Maurice combo, Paela, vegetable ravioli and dessert.

The dinner at Bertha’s restaurant is being sponsored by ThermoFisher and Shimadzu Biotech and is free of charge; however, you MUST sign up to be included. Please call Darlene at 410 955-3022 or email us at dsutton5@jhmi.edu.

Abstract:

R. L. Hettich¹, A. L. Russell¹, N. C. VerBerkmoes¹, M. Shah¹, C. Pan¹, Claire Fraiser-Liggett², J. K. Jansson³

1) Oak Ridge National Laboratory, Oak Ridge, TN, 2) University of Maryland School of Medicine, Baltimore MD,  3) Lawrence Berkeley National Laboratory, Berkeley, CA

The human gastrointestinal tract is a complex ecosystem containing a delicate balance of human and microbial cells involved in an intricate symbiotic relationship.  In general, the microbial constituency helps maintain a healthy environment and aids in the efficient digestion.  However, environmental and/or genetic factors may result in an altered bacterial composition that manifests in a diseased condition, such as Crohn’s disease.  The recent availability of genomic-based molecular technologies such as whole community genome sequencing and whole community proteomics have provided unique capabilities of profiling the compositions and activities of this microbiome without having to cultivate its membership.  We are utilizing a non-targeted, mass spectrometry-based proteomics approach to identify the microbial proteins in fecal samples from human female identical twins. Proteome samples were analyzed with technical duplicates via a multidimensional LC tandem mass spectrometric approach on a hybrid linear ion trap-Orbitrap.  The proteome measurements identified ~ 2000-4000 proteins for each sample and replicate.  Amongst the microbial genomes, Bacteroides thetaiotaomicron, Bifidobacterium longum, Bacteroides fragilis and Bifidobacterium adolescentis were the most highly identified species, as expected since these are known to be among the most abundant bacteria in the human gut.  The majority of the microbial proteins that were identified were classified into COG categories for translation, energy generation, and carbohydrate metabolism.  The latter category is especially interesting in that it reveals one of the synergistic relationships between humans and microbes in this ecosystem.  Surprisingly, a number of innate human immunity proteins were also observed, suggesting a level of human regulation of microbial abundance.  A number of abundant unknown proteins were also identified.  The results of this study demonstrate that it is possible to obtain high quality, extensive protein identifications by integrating metagenomic and metaproteomic information from matched human twin microbiomes.

*  Research support provided by the NIH-HMP Demonstration Program.  Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the U.S. Department of Energy.

Dinner: Please join the speaker and the co-chairs for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm.  Contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) to let him know you will be there for dinner.

Abstract: The ultimate goal of MS-based clinical proteomics is in-depth molecular profiling of human cancers and translation of these findings into biomarkers and assays that physicians can use on their patients. So far, the translation of proteomic findings into clinical tests has been disappointing. Innovative experimental designs coupled with high-resolution and high-accuracy shotgun proteomics for systematic profiling of human cancers may facilitate translation of proteomic findings to clinical tests. This presentation will look at results obtained from a pilot proteomic study that describes a method for cancer biomarker development using shotgun proteomics. Using renal cell carcinoma (RCC) as a model disease, this approach utilizes tissue-directed subtractive proteomics to identify tumor proteins in peripheral blood of a patient diagnosed with non-metastatic RCC. The results indicate that tumor proteins can be detected by MS in the peripheral blood of a patient diagnosed with non-metastatic cancer. To investigate the general applicability of this approach we compared entire findings of our study with previously published investigations focused on profiling RCC using MS-based proteomics. The meta-analysis revealed a subset of 70 common proteins identified in all compared investigations of which 63 (90%) were unambiguously identified in our study. These results validate the utility of described method depicting its potential to effective and systematic molecular profiling of individual human cancers/tumors. Importantly, it has great potential for creating the cancer proteome atlas that will arise from results systematically collected from analyses of individual tumors. It should reveal individual signatures and common molecular profiles resulting in discovery of driver molecules, biomarkers and pathways implicated in a given type of cancer. The atlas should yield a comprehensive, rigorous, publicly accessible proteome database that will improve our ability to diagnose and treat human cancers.

Date: Monday, September 19, 2011

Time: Dinner and Poster viewing, 6:30 pm; Short Presentations by Vendors, 7:15 pm.

Location: Shimadzu Scientific Instrument, Inc. Training Center, 7100 Riverwood Drive, Columbia, MD 21046, (directions).

Dinner: Please contact Shelley Jackson (Shelley.Jackson@nih.hhs.gov) and let him know you will be attending dinner or presenting a poster.

We extend our thanks to Dr. Berk Oktem of MassTech for his service as a co-chair for the past 2 years and welcome incoming co-chair, Dr. Peter Nemes of FDA.

Date: Monday, June 20, 2011

Time: Dinner and Poster viewing, 6:00 pm; Short Presentations by Travel Award Recipients, 7:30 pm.

Location: Shimadzu Scientific Instrument, Inc. Training Center, 7100 Riverwood Drive, Columbia, MD 21046, (directions).

Dinner: Please contact Berk Oktem by June 17 and let him know you will be attending dinner or presenting a poster.

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2011 WBMSDG Young Investigator Travel Award Recipients

Congratulations to the following recipients of the WBMSDG Young Investigator Travel Awards
Christine Jelinek, Johns Hopkins University.
Advisor: Robert Cotter. Abstract title:  Biomarkers for Cardiac Ischemia.

Joseph Strukl, University of Virginia.
Advisor:  Don Hunt.  Abstract title: Characterization of N1 – A histone chaperone involved in establishing the embryonic epigenetic state in Xenopus laevis

Sree Rayavarapu, George Washington University.
Advisor: Kanneboyina Nageraju.  Abstract title: Biomarker Discovery in Duchenne Muscular Dystrophy Using Parallel Nanotechnology Based Affinity Capture Enrichment

Speaker: Dr. Austin Yang
Greenbaum Cancer Center, School of Medicine, University of Maryland,  Baltimore, MD

Date: Monday, May 16, 2011

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center, 7100 Riverwood Drive, Columbia, MD 21046, (directions)

Dinner: Please join co-chairs Berk Oktem, Shelley Jackson and the speaker for dinner at for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm. Contact Berk Otkem (oktem@apmaldi.com) to let him know you will be there for dinner.

Abstract: Alteration of EGFR signaling is responsible for approximately 20 % of all breast cancers.   We will present a newly developed SILAC-based synthesis degradation mass spectrometry approach to determine the consequence of aberrant EGFR signaling in modulating global protein stability during the early development of breast cancer.   A robust shotgun proteomics and deep protein profiling workflow has been developed, which includes the development of several novel MS-based informatics tools to address many issues commonly associated with multiplex SILAC-based quantification and post-translational modifications.  Large-scale protein turnover and phosphoproteomics analyses suggest the change of global protein stability, in particular EGFR, is directly modulated by mTOR/STAT3 signaling pathway.

Speaker: Professor Robert J. Cotter
Middle Atlantic Mass Spectrometry Laboratory, Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD

Date: Monday, April 18, 2011

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center, 7100 Riverwood Drive, Columbia, MD 21046, (directions)

Dinner: Please join co-chairs Berk Oktem, Shelley Jackson and the speaker for dinner at for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm. Contact Berk Otkem (oktem@apmaldi.com) to let him know you will be there for dinner.

Abstract: The epigenetic code consists of methylation of DNA and modifications to histones.  The latter include methylation (mono- to tri-), acetylation, ubiquitylation, SUMOylation and ADP-ribosylation of lysine residues, methylation of arginine, and phosphorylation of serines. Acetylation is by far the most common, as the tail regions are generally hyperacetylated.  Thus, a robust quantitative approach to acetylation has been developed using the tryptic peptides from deuteroacetylated histones, and has been extended to include methylated isoforms.  MALDI time-of-flight mass spectrometry provides a facile means to assess the global changes in histone modifications, while HPLC retention times, MS/MS and high mass resolution can be used to distinguish isomeric forms, including the ability to distinguish trimethylation and acetylation.  Our approaches are used here to monitor the specific changes resulting from inhibition of deacetylases or for several methylase and/or deacetylase deletion mutants, and to study the effects of calorie restriction in yeast.  In addition, using hyperacetylated histones and time-dependent studies, it is possible to determine the specific lysine substrates for the deacetylases Sir2 and Hst2.

Topic: Five Years, Three Boys, and Some Analytical Chemistry

Speaker: Dr. Kevin Schug,  Department of Chemistry and Biochemistry, University of Texas-Arlington

Date: Monday, March 21, 2011

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center, 7100 Riverwood Drive, Columbia, MD 21046, (directions)

Dinner: Please join co-chairs Berk Oktem, Shelley Jackson and the speaker for dinner at for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm. Contact Berk Otkem (oktem@apmaldi.com) to let him know you will be there for dinner.

Abstract: For the past five years, I have been trying to “make it” as an academician in the field of analytical chemistry.  An assistant professor needs many things in order to be successful and obtain the coveted prize known as tenure.  An important component is good projects to work on.  At the outset of my academic career, my lab was focused on development of new methods for the determination and interrogation of noncovalent interactions by electrospray ionization – mass spectrometry.  Raised primarily as a chromatographer (I don’t consider chromatography to be just a fancy inlet for MS, nor do think MS is just a fancy detector for chromatography), our contribution to this area has been the incorporation of flow injection analysis and chromatographic methods to enable multiplexing binding determinations.  We have been fortunate enough to secure funding from NSF to continue developing this work, which I consider to be very fundamental in nature.  In the meantime, since we have several nice toys in the lab and appropriate expertise, we have developed collaborations with local medical schools to perform method development and trace quantitative analysis.  In this more applied regime, we have focused on advancing sample prep and reducing detection limits so that we could track levels of estrogens, metabolites, and various endocrine disruptors in a myriad of biological fluids and tissues.  We are thankful to Eli Lilly and Company for support of some of this work. More recently, we have been trying our hand at the development of an ambient ionization technique, we have termed continuous flow – extractive desorption electrospray ionization (CF-EDESI).  This technique appears to have great promise for analysis of proteins (charge-state manipulation), as well as analytes from non-ESI-friendly solvents.  Over five years, our lab has worked on a myriad of different and interesting projects.  I will present a small compilation of this work in this talk.  I have also been fortunate to have a patient and supportive wife who has provided me with three beautiful boys.  Ben (5), Luke (3), and Wes (4 months) will be your guides through this presentation (but don’t worry, I’ll be doing all the talking).

Speaker: Dr. Sandy Markey, National Institutes of Health, NIMH, Bethesda, MD

Date: Monday, February 28, 2011

Time: 7:30 pm

Location: Shimadzu Scientific Instrument, Inc. Training Center, 7100 Riverwood Drive, Columbia, MD 21046, (directions)

Dinner: Please join co-chairs Berk Oktem, Shelley Jackson and the speaker for dinner at for dinner at the Ram’s Head Tavern at Savage Mill, 8600 Foundry Street, Savage, MD 20763 at 5:30 pm. Contact Berk Otkem (oktem@apmaldi.com) to let him know you will be there for dinner.

Abstract:

Mass spectrometric instrumentation for discovery based proteomics and for targeted analyses has evolved to become useful in research for neuroscience and mental health. There have been significant research efforts directed toward the genomics of mental disorders with clear inheritability using family and twin studies. These have resulted in the identification of multiple genes associated with bipolar disorder and schizophrenia, but aberrant production or metabolism of the implicated gene protein products has not been demonstrated. Our lab has pursued several approaches to research related to these questions that will be presented: (1) label free quantification of peptides to detect proteins and networks that are responsive to dissimilar therapeutic drugs and (2) methods for stable isotope labeled peptide production for absolute quantification.