Day 2 :
Keynote Forum
Elliot R. Bernstein
Colorado State University, USA
Keynote: Euv laser mass spectrometry and photoelectron spectroscopy of mass selected neutral clusters and molecules
Time : 9:00 - 9:30 AM
Biography:
Bernstein received his Ph.D. degree from Caltech and was a post doctoral fellow at the University of Chicago. He has been at CSU since 1975 where he has studied molecular crystal vibrational and electronic excitons and phase transitions, cryogenic liquids, and gas phase clusters with a central focus on intermolecular interactions. Recently his research has focused on chemical reactions of neutral and ionic clusters. This latter research area has involved catalytic and photocatalytic cluster systems, solute/solvent systems, the reactions of ionized molecules and clusters, and initial steps in the release of stored energy molecular species.
Abstract:
A long standing set of goals for studies of systems of inhomogeneous, neutral clusters (e.g., MmXn or (molecule)p) has been to mass sort and select them individually for determination of physical and chemical properties of each neutral cluster by spectroscopic techniques. We have constructed appropriate instrumentation to achieve these important goals employing photoelectron spectroscopy (PES), driven by both visible (for MmXn -) and EUV (for MmXn0) radiation. Our 26.5 eV/photon EUV laser can ionize any neutral cluster or molecule (EUV PES) that can be identified and isolated. The algorithm includes the following steps: 1. generation of cluster negative ions in a laser ablation supersonic source with the addition, as required, of low energy electrons from a Y2O3 disk; 2. separation of these anionic clusters in a reflectron time of flight mass spectrometer (RTOFMS); 3. selection and slowing of specific, chosen clusters in a mass gate/momentum deceleration stage; 4. threshold photo-detachment of the sorted and selected negative ion clusters with a tunable VIS/UV laser to generate neutral, isolated clusters; and 5. EUV PES of these neutral clusters. Such studies generate vibrational and structural information on the ground states of the neutral clusters (through VIS/UV PES), and information on the ion states of the clusters (through EUV PES). The presentation will include PES results on various metal oxides, sulfides, and other cluster systems and molecules.
Keynote Forum
Brigitte Simons
SCIEX, Canada
Keynote: Quantitation of Lipids in High Throughput; the shotgun lipidomics profiling of NAFLD and steatohepatitis by ion mobility-MS data acquisition techniques
Time : 9:30 - 10:00 AM
Biography:
Brigitte Simons is a market development specialist at SCIEX, specializing in metabolomics using accurate mass time-of-flight mass spectrometry solutions. Prior to working at SCIEX, Brigitte received her Ph.D. in Chemical Biology at the University of Ottawa. She then completed two research post-doctoral fellowships at the Centre for Biologics Research at Health Canada and the National Institute of Heart Lung and Blood in Bethesda MD. Brigitte also spearheads an academic partnerships program for our North American business – which listeners can browse sciex.com for more information or contact Brigitte for more information.
Abstract:
This presentation will address technologies that directly address clinical research experimentation of lipids in tissue extracts, keeping sampling and data processing throughput in mind. The human lipidome contains >100,000 different molecular species found within a small mass range; consequently, isobaric overlap makes unambiguous identification and quantitation of lipid species difficult. Herein, methods that utilize high sensitivity MS/MS techniques using a high sensitivity triple quadrupole linked LIT mass spectrometer to isolate lipid classes for identification of molecular composition and quantitation have been investigated. Furthermore, QqQ platforms can now be coupled to differential ion mobility (DMS) devices and have shown to resolve phospholipid sub-classes, triglycerides, and strikingly, the sphingomyelins (SM) were resolved from PC molecular species. The latter observation is significant considering these two classes cannot be resolved using triple quadrupole strategies alone, and their masses overlap significantly. Alterations in hepatic phospholipid composition, especially the amount of phosphatidylcholine (PC) and the ratio of PC to phosphatidylanolamine (PE) might contribute to the development non-alcoholic fatty liver disease (NAFLD). Aim was to compare PC/PE ratio in patients with NAFLD or chronic Hepatitis C (CHC) to healthy controls. This was a cross-sectional study. Liver samples were obtained from healthy controls (HC), patients with simple steatosis (SS), and steatohepatitis (NASH). Lipids were extracted from liver tissue and the total PC and PE lipid detected was quantified by tandem mass spectrometry using multiple precursor ion scanning [normalized by internal standards]. Hepatic PC/PE ratio was lower in all 3 patient groups, mainly due to a reduction in PC, which was significant in SS and NASH compared to controls. This study confirms previous animal data on PC/PE in NAFLD and represents a high throughput lipidomics platform’s utility to clinical outputs.
Keynote Forum
Magnus S. Magnusson
University of Iceland, Iceland
Keynote: Similarity of Hierarchical Structured Clustering in Human and Neuronal interactions and on DNA: Structural and Functional analogies.
Time : 10:00 - 10:30 AM
Biography:
Magnus S Magnusson, Research Professor. PhD in 1983, University of Copenhagen. Author of the T-pattern model and the corresponding detection algorithms in THEME. He has focused on real-time organization of behavior, co-directed DNA analysis, numerous papers and invited talks at numerous conferences and universities in Europe, USA and Japan. Deputy Director 1983-1988, Anthropology Laboratory, Museum of Natural History, Paris. Repeatedly invited temporary Professor at the University of Paris. Since 1991, Founder and Director of the Human Behavior Laboratory, University of Iceland. Since 1995, he is in collaboration between 24 universities based on “Magnusson’s analytical model” initiated at the Sorbonne, Paris
Abstract:
Hierarchical structured clustering (HSC) seems characteristic of the structure of the universe balancing a small number of forces, some pulling others pushing apart, the self-similar fractal distribution of matter in the universe thus reflecting HST rather than just dispersion or clumping. HSC also characterizes a proposed pattern type, called T-pattern, detected in the temporal organization of many kinds of verbal and non-verbal human, animal and neuronal behavior and interactions and is also characteristic of the structure of DNA. Functional analogies seem to exist between the occurrence of T-patterns in “cell city” and in human cities. Structural self-similarity over many levels of biological organization suggests the possibility of a unified (mathematical, bioinformatics and system biological) approach. The T-pattern is described as a repeated hierarchical and self-similar tree structure based on a single non-terminal relation, called a critical interval (CI) relationship. The instances of a T-pattern may be seen as repeated statistical pseudo-fractal objects characterized by statistically significant translation symmetry. THEMEtm (by M.S. Magnusson, ©PatternVision Ltd) is special purpose T-pattern detection and analysis PC software using a special CI detection algorithm combined with an evolution algorithm, presented here together with results from the analysis of behavior and interactions. From the relatively slow time scale of human and animal interactions to the much faster interactions within populations of neurons in living rat brains. Analogies are discussed between T-pattern structure and functions in the „cities of proteins“ (cell city) and human cities, especially regarding specialization and the particular case of religious behavior.
- Track 1: Applications of Mass Spectrometry Track 3: New Approaches in Mass Spectrometry Track 6: Mass spectrometry Imaging Track 8: Ionization Techniques
Chair
Elliot R Bernstein
Colorado State University, USA
Co-Chair
Ioana NUTA
Univ. Grenoble Alpes, SIMAP, France
Session Introduction
Yujun Shi
University of Calgary, Canada
Title: Development of Laser Ionization Methods and Its Application in the Investigation of Chemical Vapor Deposition Chemistry
Biography:
Yujun Shi obtained her PhD in Chemistry from the University of Western Ontraio (now Western University) in Canada. She did postdoctoal work at the National Research Council (NRC) of Canada in Ottawa on an NSERC Visiting Fellowship. She is currently an Associate Professor in the Department of Chemistry at the University of Calgary. She has published more than 50 papers in referred journals and has been serving as an editorial board member for Canadian Journal of Chemistry (Associate Editor) and Frontiers in Physical Chemistry Chemical Physics.
Abstract:
Laser ionization mass spectrometry is a powerful diagnostic tool for a variety of applications including biomedical research, drug delivery, atmospheric chemistry, and chemical vapor deposition chemistry. Our laboratory has developed several complimentary ionization methods, including the non-resonant single photon ionization (SPI) with a vacuum ultraviolet (VUV) laser radiation of 118 nm (10.5 eV) and the laser induced electron ionization (LIEI) for the diagnosis of the decomposition chemistry of precursor molecules in the process of hot wire chemical vapor deposition (HWCVD). In this work, results from our investigation on the decomposition of orgaosilicon molecules on heated tungsten and tantalum wires will be reviewed. A common decomposition pathway of forming methyl radicals has been demonstrated for methy-substituted silane molecules and methyl-substituted four-membered-ring silacyclobutane compounds. The formation mechanism and the activation energy for this pathway will be presented in this work. For the four-membered-ring organosilicon precursors, additional decomposition channels were also found from the ring-opening reactions to form alkene and silenes. The investigation of the decomposition chemistry of organosilicon molecules on heated metal wires provides helpful and important insights in understanding the secondary gas-phase reactions in HWCVD processes and also the surface reactions on the metal catalysts.
Andrew Ewing
University of Gothenburg and Chalmers University of Technology, Sweden
Title: Mass Spectrometry Imaging in Flies, Cells, and Vesicles
Biography:
Andrew Ewing received a PhD from Indiana University. After 25 years at Penn State University, he is now Professor at Chalmers University and University of Gothenburg, Sweden. His 300 publications have been cited over 15000 times with an H-index of 69. He has recently received the Norblad-Ekstrand Medal of the Swedish Chemical Society (2014), and the Pittsburgh Conference Award in Analytical Chemistry (2015). He is an Honorary Professor at Nanjing and Beijing Universities of Science and Technology. He is a member of the Royal Swedish Academy of Sciences (from 2012) and the Gothenburg Academy of Arts and Sciences (2013).
Abstract:
We have been developing mass spectrometry imaging methods to study the process of neurocommunication at the system and cellular level. We focus on PC12 cells as a model of exocytosis and the fly model (Drosophila melanogaster) providing a unique system to examine neurotransmitter release and drug dependence mechanisms in a small, but complete system. Mass spectrometry imaging with ion beams allows spatial resolution of a few micrometers down to 40 nanometers in favorable cases. We have been using secondary ion mass spectrometry (SIMS) with a unique 40-kV argon cluster ion source and the NanoSIMS to measure the lipids across the fly brain and catecholamine in nanometer vesicles, respectively. Here, we have focused on the effect of the drug, methylphenidate, on lipid composition in the brain and find that it varies in a way that might affect learning and memory. We have also used NanoSIMS to measure transmitter in subregions of nanometer vesicles. Combined with other new methods to measure the content of the interior of vesicles, we have begun to investigate the details and implications of open and closed exocytosis on regulation of how the brain works.
Libong Danielle
Paris South University, France
Title: Identification of lipid biomarkers of atopic skin by normal phase chromatography/high resolution mass spectrometry
Biography:
Danielle Libong has completed her PhD at the age of 28 years from Ecole Polytechnique of Palaiseau (France). She is senior lecturer in the Interdisciplinary Academic Unit Lip(Sys)² Lipids (Analytical and Biological Systems) of the School of Pharmacy of Paris-South University. Lip(Sys)² is a research team in analytical chemistry with an important experience in lipid analysis by chromatographic, hyphenated methods and optical spectroscopy. She has published more than 25 papers in reputed journals.
Abstract:
The strategy used to determine the lipid biomarkers involve lipidomics analysis of normal and atopic skin. In the case of skin called atopic, permeability of the epidermis or the stratum corneum is a problem. Stratum Corneum has been described as a type of “brick and mortar”, in which the corneocytes are bricks. Extracellular spaces are filled with lipids that are divided into three main classes: ceramides, fatty acids and cholesterol. Ceramides constitute a major class of lipids and are composed of a sphingoid and a fatty acid moiety. They play a major role in skin permeability. Their organization is essential to maintenance of the barrier function. Lipidomics offers a unique opportunity to analyze the complex role of lipids in cellular processes. The first step focuses on the development of a method for separation and detection of lipid classes molecular species using (Normal Phase) LC/ MS in high resolution. Different ionization modes have been tested between APPI and APCI. The main goal is to collect information from all detected peaks, the profile of the sample provides the relative distribution of species and the molecular mass of each. Statistical comparison of the profiles obtained for several samples of both populations reveal the characteristic signals of over / under expressed signals in each group. In the end we characterize and identify biomarker by MS/MS.
Ioana NUTA
Univ. Grenoble Alpes, SIMAP, France
Title: Mass spectrometry with Knudsen cell applied to the investigation of organometallic precursors vapors
Biography:
Ioana NUTA received her Ph.D from Orleans’s University (France) in 2005 working on fluoride molten salts baths at high temperature by NMR spectroscopy in CNRS-CEMHTI Laboratory. She then completed a post-doctoral fellowship at ENSI Caen (2006) in CNRS-LCS Laboratory and at BRUKER (Germany) in department of NMR probe devellopement for catalytic studies. Since 2007, she joined CNRS- SIMAP Laboratory as reasercher in “Thermodynamic and Process Optimization” team where she has in charge the themodynamic studies on gaseous phase using Knudsen effusion Mass Spectrometery. Current studies focus on thermodynamics of organometallics, molten salts and oxides.
Abstract:
A special Knudsen cell reactor coupled to a mass spectrometer, was specifically designed for the study of organometallic precursors. This reactor is built as tandem cells: an evaporation cell and a cracking cell. This reactor tries to simulate the conditions found in the bubbler of the ALD system by the evaporation cell and in the hot reaction zone of ALD by the cracking cell. The first stage reactor - an evaporation cell - provides an input saturated vapor flow operating from room temperature to 333 K. The second stage cell, named cracking cell, operated from 333 to 723K in the present study. During experiments, the effusion orifice is externally opened for direct mass spectrometric measurements of saturated vapour pressures. The device has been tested using the well-known mercury system. The thermal cracking of the gaseous precursor pentakis dimethylamino tantalum (PDMAT), generally adopted in the ALD/CVD TaN deposition processes, has been studied in the temperature range from 343 to 723K. Experiments showed the apparition of many gaseous species when cracking temperature increased and in particular the dimethylamine, HNC2H6 (g), corresponding to the saturated organic branches of PDMAT. Decomposition products of the HNC2H6 branch were observed at relatively high temperature, namely above 633K. This gas phase study - as for the saturated preceding one- shows the presence of oxygen containing molecules in PDMAT cracked vapor. Thus it allows explaining the systematic presence of oxygen contamination in the deposited TaN films observed in ALD/CVD industrial processes.
Stefanie Maedler
Ontario Ministry of the Environment and Climate Change, Canada
Title: Ultra-trace level speciated isotope dilution measurement of Cr(VI) using ion chromatography tandem mass spectrometry in environmental waters
Biography:
Stefanie Maedler received her Ph.D. in analytical chemistry in 2011 from the Swiss Federal Institute of Technology in Zurich, Switzerland (ETHZ). After 2 postdoctoral fellowships at York University and University of Toronto, Canada, Dr. Maedler joined the Ontario Ministry of Environment and Climate Change as a Development Scientist. She has 10 years of expertise in the use of mass spectrometry (MS) for biological and environmental applications and is currently focusing on the speciation of inorganic analytes and understanding their toxicological effects on fish through MS-based metabolomics approaches.
Abstract:
The reliable analysis of highly toxic hexavalent chromium, Cr(VI), at ultra-trace levels remains challenging, given its easy conversion to non-toxic trivalent chromium. The new approach demonstrates a novel analytical method to quantify Cr(VI) at low ng/L concentration levels in environmental water samples by using speciated isotope dilution (SID) analysis and double-spiking with Cr(III) and Cr(VI) enriched for different isotopes. Ion chromatography tandem mass spectrometry (IC-MS/MS) was used for the analysis of Cr(VI) as HCrO4- → CrO3-. While following a classical linear multipoint calibration curve a method detection limit (MDL) of 7 ng/L Cr(VI) was achieved, the modified SID-MS method adapted from U.S. EPA 6800 allowed for the quantification of Cr(VI) with an MDL of 2 ng/L and provided results corrected for Cr(VI) loss occurred after sample collection. The adapted SID-MS approach proved to yield more accurate and precise results than the multipoint calibration method, allowed for compensation of Cr(VI) reduction during sample transportation and storage while eliminating the need for frequent external calibration. The SID approach permitted continuous sample analysis for several days without the need for recalibration. This new developed IC-MS/MS method represents an alternative to the routinely used inductively-coupled plasma (ICP) instrumentation, IC-ICP-MS, and offers several advantages over detection with ICP-MS for Cr(VI), such as the absence of polyatomic interferences of 52Cr formed in the ICP (36Ar16O, 40Ar12C, 35Cl16OH, and 37Cl14NH) that require the use of dynamic reaction/collision cells or high-resolution double-focusing sector field instruments.
Nhu Phan
University of Gothenburg, Gothenburg, Sweden
Title: Lipid imaging of invertebrate model systems by secondary ion mass spectrometry
Biography:
Nhu Phan has been working on her PhD at the University of Gothenburg, Sweden. Her research is about developing mass spectrometry imaging methods for biological applications on different biological models, from single cell imaging to invertebrate models particularly C. elegans and Drosophila. One of her main researches is studying the effects of administrated stimulant drugs on the neurochemistry of Drosophila brain. The significant finding in this study is that the stimulant drug induces lipid structure of the brain and that the brain lipids could closely relate to learning and memory.
Abstract:
Drosophila melanogaster and C. elegans (flies and worms, respectively) are common biological model systems, which have relatively simple anatomy and behavior but possess highly conserved molecular and cellular processes compared to humans. We have applied time of flight secondary ion mass spectrometry (ToF-SIMS) to study lipid structural effects of stimulant drug methylphenidate on the fly brain, and to investigate the 3D chemical anatomy of C. elegans. Different distributions of various biomolecules, particularly fatty acids, eye pigment, diacylglycerides, phospholipids have been found across the fly brain. Lipid structures, particularly diacylglycerides (DAG), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositiol (PI), are shown to dramatically alter following the administration of methylphenidate. For C. elegans, the entire worm and worm sections were imaged using 3 dimensional (3D) and 2 dimensional (2D) approaches, respectively. Significant changes in the chemical distribution were observed along the depth of the worm. In addition, correlation between 2D and 3D ion images showed different molecular structures across the worm, possible localization of the nerve ring, the cuticle, and the fluid containing space inside the worm. In addtion, we successfully used tandem MS on ToF-SIMS with a high energy 40 keV Ar4000+ gas cluster primary ion beam (GCIB) to elucidate the structures of molecular lipids in the fly brain and the worm. ToF-SIMS imaging shows great potential to elucidate chemical distributions in small invertebrate systems in relation to endogenous and exogenous effects.
- Track 1: Applications of Mass Spectrometry Track 5: Recent Advances and Development in Mass Spectrometry Track 9: Mass Spectrometry Configurations and Techniques Track 12:NMR Spectroscopy and NMR in biomedicine
Chair
Lianming Wu
GlaxoSmithKline, USA
Co-Chair
Medicharla V. Jagannadham
CSIR-Centre for Cellular and Molecular Biology, India
Session Introduction
Paul Lee
Discovery Technologies, Amgen Inc., USA
Title: High-throughput metabolic stability assays using RapidFire and Accurate-Mass Spectrometry
Biography:
Paul Lee joined Amgen Inc. in 2004. He currently leads a group of scientists and is responsible for developing biochemical and cellular assays, implement high-throughput screens and lead discovery in the early drug discovery process. Prior to joining Amgen, Paul held various positions at Pfizer, Pharmacia, and Glaxo-Wellcome for over 12 years. Paul received his Ph.D. in Pharmacology from University of Hong Kong and completed his postdoctoral training at NIH.
Abstract:
Metabolic stabilities of small molecules, in the presence of liver microsomes and/or hepatocytes, have been frequently assessed during lead optimization in the early drug discovery process. The resultant ranking is a key selection criterion for compound advancement into in vivo studies because a more metabolically stable compound will likely give rise to a longer half-life in animal studies. In vitro metabolic stability studies have been routinely performed using pooled liver microsomes from various species (human, monkey, dog, rat and mouse). The key detection technology of this assay is by monitoring the turnover of the compound in the presence of liver microsomes using liquid chromatography-tandem mass spectrometry (LC-MS). However, the bottleneck of the assay falls on the long separation time in LC. The use of RapidFire, a system consisting of a solid-phase extraction column, automated plate handler, sample injector, and software interface with MS, has significantly improved the throughput. In addition, the use of an Accurate-Mass Quadrupole Time-of-Flight (QTOF) MS system with RapidFire eliminates the need of tuning prior to sample analysis and further streamlines the MS analysis process. In this report, we describe the implement of RapidFire-QTOF system in human and rat metabolic stability assays, resulting in a throughput ten times higher than the traditional LC-MS method without compromising data quality.
Lianming Wu
GlaxoSmithKline, USA
Title: Ambient Mass Spectrometry: New Directions in Pharmaceutical Analysis
Biography:
Lianming Wu obtained a Ph.D. in mass spectrometry (MS) from Purdue University under the guidance of Prof. R. Graham Cooks and followed by Postdoc Research Fellow in MS-based proteomics at Pacific Northwest National Laboratory under the guidance of Dr. Richard D. Smith. Dr. Wu has published forty one peer-reviewed scientific papers and book chapters on the subject of mass spectrometry and gave a number of oral presentations in national/international conferences such as ACS National Meeting and PittCon. He is currently working in Global Spectroscopy, Analytical Sciences & Development, GlaxoSmithKline.
Abstract:
Ambient mass spectrometry (AMS), that allows rapid chemical analysis of untreated samples in the ambient environment, represents novel directions in pharmaceutical analysis. Because AMS is performed by direct sampling/ionization of analytes from native samples, it allows high throughput for the analysis of compounds in matrices using solvent-based desorption methods such as desorption electrospray ionization (DESI) or plasma-based methods such as direct analysis in real time (DART). As one of the most widely studied ambient ionization methods, DESI uses fast-moving solvent droplets to extract analytes from surfaces and propel the resulting secondary microdroplets towards the mass analyzer. The development of AMS takes advantage of the knowledge and experience accumulated in mass spectrometry as well as other areas of chemistry, physics and engineering. Several applications of AMS in supporting pharmaceutical product development are presented: (1) direct analysis of samples in matrix with little to no sample pretreatments; (2) MS-imaging (drug mapping in biological samples and tablet formulation as well as diagnosis of tissues); (3) conformation study of proteins and oligonucleotides; and potentially (4) investigation of salt forms of drugs in tablet or capsule formation. Manipulation of the chemistry accompanying ambient ionization can be used to further optimize chemical analysis. Extensions of the methodology to pharmaceutical analysis (such as direct analysis of biotransformation reaction mixtures in biological matrixes and trace analysis of genotoxic impurities) by reactive DESI via in-situ chemical derivatization (viz. gas-phase ion/molecule reactions) on surface are also covered.
Suneetha Achanti
Department of Pharmaceutical Analysis, Hindu College of Pharmacy, India
Title: PDAS–MSD- LC MS- Prescriptive Drug Analysis Study in Rat Plasma for Multiple Sclerosis Disease by LC-ESI/MS
Biography:
Dr. A. Suneetha, Professor & Head, Department of Pharmaceutical Analysis at Hindu College of Pharmacy, Guntur, A.P since 2001. She has completed her Ph.D at the age of 33 years from Acharya Nagarjuna University, Nagarjuna nagar. She has total 16 years of teaching and research experience. She has published more than 40 research papers in various reputed National and International journals. She served as BOS member of ANU in Pharmacy during 2010 to 2013. She is recognized as Research Supervisor from Acharya Nagarjuna University for guiding PhD scholars and she is a life member of APTI.
Abstract:
Prescribing a single drug and its administration is not sufficient in neuro diseases like multiple sclerosis. Combination therapy is growing enormously to decrease the number of medications for a single disease or their associated diseases. In clinical research estimation of concomitant drugs plays a key role to study the drug-drug interactions. The research in the current article has undertaken to provide an accurate method for evaluate the pharmacokinetic parameters of Carbamazepine (CBZ), Duloxetine (DLX), Tamsulosin (TSL) and Teriflunomide (TFM) using Doxofylline (DXF) and Ibuprofen (IBP) as internal standards (IS) in rat plasma by LC- MS when used as combination therapy. The developed bioanalytical method has been validated according to ICH guidelines. The obtained LODs and LOQs of all the drugs were adequate and may useful to perform the pharmacokinetic study in rat plasma. Based on the results, we can conclude that the present method is suitable for quantification of multiple analytes simultaneously without any interference and matrix effects. The concomitant drug analysis along with the target analyte is more advantageous than single compound analysis and also useful in drug interaction and toxicology studies. This method can also be useful in estimating the plasma samples of patients who administer these drugs.
Sermin Tetik
University of Marmara, Turkey
Title: Mass Spectrometry Technology for Biomarker Discovery
Biography:
Dr. Sermin Tetik is completed her PhD degree at the 2003 years at University of Marmara (Istanbul-Turkey). She have been working an academician at the same University, Faculty of Pharmacy in Istanbul-Turkey as associated professor since five years and project director of her research team focusing on thrombosis -hemostasis area. She has published more than 30 papers and above 50 international abstract in reputed journals and conferences, serving as an editorial board member of repute.
Abstract:
Human tissues and organs have affected depending on pathological condition of diseases. Proteins are basic functioning molecules of cells accordingly to modification intensities. Protein modification can be differences in cellular interaction, localization, activity, protein concentration, and co-/posttranslational. Protein biomarker discovery has covered some subtitles or points such as differantially expressed proteins for disease specific protein isomers and abnormal protein activity. Proteomic analysis and conclusions have allowed the potential protein biomarker discovery and thus diagnosis of disease and prognosis can be possible. MS-based proteomic has correlated between protein modification and a certain disease platform to accurately and confidently measure the proteins in sera or plasma which are low-abundance concentration. MS-based quantitative platform use for tissue samples or several body fluids of protein biomarker discovery. Similar to whole tissue samples human fluids such as gastric fluid, urine, blood have also been evaluated in the last decade studies. Intensity of modifications of proteins as post translational can also be indicative of a disease or its progression. Activity-based protein platform changes have allowed for proteome researchs of enzymatic and protein-drug interaction events.
Matthew B. O’Rourke
University of Technology Sydney, Australia
Title: Improving reproducibility and resolution in Imaging Mass Spectrometry (IMS) through the application of low cost manual sample preparation techniques
Biography:
Matthew O’Rourke currently has his PhD in submission after 2 years of candidature and is a resident at the university of Technology in Sydney Australia. He has published several papers in the field of Imaging mass sectrometry and tissue preservation and is an expert in MALDI applications. He also holds a patent in Australia for the development of proprietary MALDI based technology for the identification of microorganisms.
Abstract:
Matrix assisted laser desorption ionisation imaging mass spectrometry (MALDI IMS) is a technique that is currently seeing widespread use in both research and diagnostics for the spatial analysis of a range of biomolecules. It is currently predominantly used for the diagnostic analysis of cancer biomarkers in tissue microarrays. This requires automated sample preparation methods that need to be highly reproducible. Automated preparation methods rely on the spraying of dissolved matrix that can cause delocalisation of molecules, limiting potential image resolution to that of the smallest possible droplet size (usually >20 µm). The instruments are also expensive to purchase and require regular maintenance and servicing by qualified engineers. We therefore describe an alternate sample preparation method that is both low cost and highly reproducible, utilising sublimation and vapour recrystallisation. This eliminates the issue of delocalisation of molecules and allows for a potential resolution of ~1 µm, thereby enabling highly reproducible, ultra-high resolution images when the mass spectrometry instrumentation that can acquire at sub- µm resolution becomes available.
Medicharla V. Jagannadham
CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
Title: Chemical modification of peptides followed by mass spectrometry: Implications in proteomics
Biography:
M.V. Jagannadham is working as a senior principal scientist and project leader at the Centre for Cellular and Molecular Biology. He published 45 research papers in internationally reputed scientific journals. He trained several students, conducted meetings and workshops in proteomics. He received “Bharat Jyothi” award from India International Friendship Society, New Delhi in 2014 and “Eminent Mass Spectrometrist” award from the Indian Society for Mass spectrometry (ISMAS) in 2015. His current research Interests are proteomics, particularly in improving the de novo sequencing efficiency of peptides using MS techniques, structural and functional studied of outer membrane vesicles of Gram-negative bacteria.
Abstract:
Determination of the sequence of the peptides, obtained from the proteolysis digestion of a protein, using mass spectrometry, is a crucial requirement in proteomics. Sequence of the tryptic peptides is usually obtained by database search or by de novo sequencing. The poor spectral quality and signal to noise ratio interferes in the analysis. Different types of algorithms (MASCOT, SEQUEST and others) are used for interpretation of the mass spectral data. When the database is not available, de novo sequencing is the only way to determine the sequence. The de novo sequence can be obtained by employing different methods of fragmentation (CID, HCD, ETD and ECD), getting information on their sequence and combining this information. Even with MS instruments with high mass accuracy and speed of analysis, the reproducibility of the mass spectrometry-based proteomics is being questioned from time to time. Acetylation of peptides improved the spectral quality, exhibited by an increase in b ion intensities in the MS/MS spectra, improves the efficiency of de novo sequence and helped in validating the database search results. It is a simple reaction, which can be carried out on complex protein digests as is required in proteomics. The identification of proteins from an Antarctic bacterium Pseudomonas syringae Lz4W and other species using this strategy will be discussed.
Adrijana Leonardi
Jozef Stefan Institute/Department of molecular and biomedical sciences, Slovenia
Title: Snake venomics and antivenomics of the nose-horned viper, Vipera ammodytes ammodytes
Biography:
Adrijana Leonardi is a researcher at the Department of molecular and biomedical sciences at the Jozef Stefan Institute. She is a biochemist with a deep research interest in toxicology, protein structure and structure–function relationships. She has been studying hemostatically active components of snake venoms and other medically interesting compounds from animal venoms. Recently she started collaborations with research groups studying the interactions of nanoparticles with plasma proteins. She is mentoring graduate and undergraduate students. Her work was published in 36 papers in reputed journals.
Abstract:
Snake venoms are a deep well of natural compounds with high specificity for various biological targets, from which we draw new pharmacological leads. The nose-horned viper, Vipera a. ammodytes, is the most venomous European snake inhabiting a large part of the south-eastern Europe and Asia Minor. Hemorrhage and coagulopathies are the most pronounced effects of envenomation in humans. We used snake venomics approach, combining proteomics and transcriptomics, to survey the protein composition of the venom with the aim of discovering new pharmacologically active substances. Proteins were separated by two-dimensional polyacrylamide gel electrophoresis into 208 spots and identified by mass spectrometry using the venom gland cDNA library sequence data. In an activity-guided fractionation, hemostatically active components, effecting blood coagulation and platelet aggregation, were detected and isolated: procoagulants metalloproteases - factor IX, X and prothrombin activators, anticoagulant fibrinogenolytic serine proteinases with kallikrein-like and angiotensin I-degrading activity, C-type lectin-like proteins inhibiting vWF-dependant platelet aggregation, dimeric disintegrins inhibiting vWF-, collagen- and ADP-dependant platelet aggregation and phospholipases A2 with anticoagulant and platelet anti-aggregation activities. The only adequate and efficient treatment of snakebite envenoming is serotherapy. The term “antivenomics” describes proteomic procedure of identification of those polypeptides in the venom which possess epitopes that are weakly or not at all recognized by the standard homologous antivenom. Using antivenomics approach we aimed to deepen and broaden our efforts towards formulation of the optimal antigenic composition of the sample for immunization of animals to prepare highly effective antiserum for immunotherapy after envenomation with the nose-horned viper venom.
Athula B. Attygalle
Center for Mass Spectrometry, Stevens Institute of Technology, USA
Title: Charge-site location and structure elucidation of gaseous ions
Biography:
Attygalle is internationally recognized as a researcher and an educator of mass spectrometry and chemical ecology. He has extensive experience in conducting hands-on laboratory workshops and lectures on mass spectrometry. He has conducted mass spectrometry workshops in Malaysia, Pakistan, Brazil, Costa Rica, the Dominican Republic, Switzerland, Saudi Arabia, Germany, and India. Dr. Attygalle has published nearly 200 peer-reviewed research articles. Research reported in some of his joint publications has been singled out for coverage on numerous occasions by the media including the New York Times. Before joining Stevens, he served as the Director of the mass spectrometry facility of Cornell University from 1988 to 2001.
Abstract:
The location of the charge site is an important prerequisite to interpret fragmentation spectra of ions. Although the challenge appears seemingly simple, in reality it is not at all a trivial problem. One often assumes that the charge location can be easily predicted by knowing the gas-phase acidity or basicity of various groups present in a polyfunctional molecule. However, in reality this a very challenging problem because generalizations valid for solution-based chemistry cannot be extended directly to rationalize gas-phase phenomena. For example, there is sufficient experimental evidence, particularly from ion mobility mass spectrometry, to demonstrate that deprotonated p-hydroxybenzoic acid could exist in gas phase as mixture of carboxylate and phenoxide forms. Analogously, protonated aniline exists as mixture of nitrogen- or ring-protonated forms. Sometimes, the incipient proton that imparts the charge to a molecule is mobile. For example, methyl dihydrocinnamate initially protonates on the carbonyl group. Then the charge ambulates to the phenyl ring before finally migrating to the methoxy oxygen atom to afford a subsequent loss of methanol. Analogoulsy, The charge-imparting proton migrates to the ring before a molecule of water is eliminated from protonated benzoic caid. Using many similar examples, the methods, which include ion-molecule reactions, H/D-exchange studies, ion-mobility spectroscopy, and isotope-labeling experiments, available to elucidate intricate ion-fragmentation pathways will be presented and discussed.