Day 2 :
Keynote Forum
Mikhail Y Golovko
University of North Dakota, USA
Keynote: Tissue lipidomics analysis of eicosanoids using tissue microwave fixation and chiral LC-MS/ MS quantification
Time : 10:00-10:45
Biography:
Mikhail Y Golovko obtained his PhD at Tver State Medical University, Russia. He is currently an Associate Professor in the Department of Biomedical Sciences, University of North Dakota (UND), USA.He joined UND in 2003 post completion of his PhD. His research interests are focused on brain lipid metabolism under neurodegenerative conditions including Alzheimer's disease and stroke.
Abstract:
A number of tissue bioactive lipids, including eicosanoids, are dramatically and instantly increased during tissue removal from the body and handling, thus altering quantification results for their basal levels. In addition to enzymatic synthesis, eicosanoid like isoprostanes are also produced upon oxidative stress. We, and others, have demonstrated that high energy focused microwave irradiation (MW) prevents a rapid, 30-fold increase in brain and 150-fold increase in kidney eicosanoid mass within seconds upon tissue extraction from the body. In addition, MW is also required to prevent postmortem alterations of another bioactive lipid group, endocannabinoid, including 2-arachydonoylglycerol (2-AG) and N-arachidonoylethanolamine. We validated stability of endogenous eicosanoids and endocannabinoids under tissue exposure to MW and the application of MW to measure true basal levels of prostaglandins, isoprostanes, and endocannabinoids. We also demonstrated that chiral chromatography is required to differentiate between eicosanoids and non-enzymatically produced isoprostanes. Our results indicate that MW combined with chiral LC-MS/MS is a safe and required technique to quantify tissue levels of a number of bioactive lipids including eicosanoids.
Break: Networking & Refreshments 10:45-11:05 @ Europa Foyer
Keynote Forum
Eloiza H Tajara
University of Sao Paulo, Brazil
Keynote: Plasma metabolomics for identifying diagnostic and prognostic biomarkers of oral cancer
Time : 11:05-11:50
Biography:
Eloiza H Tajara obtained her Bachelor’s Degree in Biological Sciences (1969) and PhD Degree in Genetics (1980) at the University of São Paulo, Brazil. She did her Postdoctoral studies at the Southwest Biomedical Research Institute (Arizona, USA) and was a Fellow of the Union for International Cancer Control in the Eleanor Roosevelt Institute for Cancer Research (Colorado, USA) in 1994 and in the University of Chicago (USA) in 1997. She is currently an Associate Professor at the Faculty of Medicine of São José do Rio Preto, Sao Paulo, Brazil. She is the Coordinator of GENCAPO (Head and Neck Genome Project), a consortium of research groups from universities and hospitals in the State of São Paulo, in collaboration with researchers from the Federal University of Espírito Santo and the International Agency for Research on Cancer, Lyon, France whose goal is to develop clinical, epidemiological and molecular analysis of head and neck cancer. She has experience in Genetics and Molecular Biology, with emphasis on Cancer. Her scientific papers were cited 3435 times and the H index is 28.
Abstract:
Metabolomics has proved to be an important tool in cancer research, it allows the identification of molecular pathways behind the tumor phenotype and clinically useful markers. In fact, the data have shown the potential of metabolites to classify different types of cancer, both in diagnostic and prognostic levels. In head and neck cancer, several metabolomic data have been published in biofluids and tumor tissues. In the present study, plasma analysis was performed to identify metabolic profiles in oral squamous cell carcinoma (OSCC) patients and controls as well as in patients at different stages of disease. 1H NMR (Proton Nuclear Magnetic Resonance) analysis was performed on plasma samples from 47 OSCC cases and 49 controls to investigate association of metabolite concentrations with clinical, pathological and lifestyle variables. The results showed that clinical and lifestyle variables affect the plasma concentration of metabolites, especially those involved in metabolic pathways related to energy homeostasis. Mass spectrometry analysis of plasma samples was also performed to compare metabolite concentrations of 61 OSCC patients with 61 controls and of different cancer stages to obtain an insight on the dynamics of metabolite changes during oral neoplastic progression. Identification of the metabolites with significantly different concentrations was carried out using the PLS-DA (Partial least squares Discriminant Analysis) method. The 15 top features with higher VIP scores included acylcarnitine, serotonin and phosphatidylcholines more expressed in plasmas from patients. In addition, the data showed a deregulation in
carnitine concentrations and an increased biosynthesis of unsaturated lipids during neoplastic transformation, but more evident in the stage with larger tumor volume. In summary, the OSCC patients exhibited a distinct plasma metabolic profile suggestive of abnormal lipogenesis and energy metabolism, which is apparent in advanced stages of the disease and may contribute to proliferation and inflammation. Such signature, if used in monitoring tests, may contribute to prognosis and treatment of OSCC.
- Metabolomics in Drug Discovery | Cancer Metabolomics | Transcriptomics and Proteomics
Location: Athens
Chair
Philip J Jackson
University of Sheffield, UK
Co-Chair
Eloiza H Tajara
University of São Paulo, Brazil
Session Introduction
William Hidalgo
Industrial University of Santander, Colombia
Title: Transcriptomic and metabolic analysis of the plant hormones signalling pathways in the resistant “Calcutta 4†and susceptible “Williams†banana genotypes during the interaction with the pathogenic fungus Pseudocercospora fijiensis
Time : 12:35-13:05
Biography:
William Hidalgo pursued Master of Science Degree in Biotechnology from National University of Colombia (Colombia). He is a Chemist from University of Nariño (Colombia); Doctor in Natural Science from Friedrich Schiller University Jena (Germany). Since March 2017, he joined the Industrial University of Santander (Bucaramanga, Colombia) as an Assistant Lecturer and Researcher in the Chemistry School. His scientific background is focused on the study of plant chemical defenses especially in the biological system “Musa-Mycosphaerella fijiensis” (causing the Black Sigatoka Disease in Banana plants), biosynthesis of natural products (by using 13C-tracer) and metabolomics studies by using NMR and MS spectrometry techniques. He has participated in several international conferences organized mainly by the International Society of Chemical Ecology and Latin American Association of Chemical Ecology, with a scientific contribution of ten international publications.
Abstract:
Statement of the Problem: Bananas (Musa spp.) represent one of the most important crops throughout the world. Black Sigatoka disease (BSD), caused by the ascomycete fungus Pseudocercospora fijiensis, is still one of the main phytosanitary problems facing the crop.
Methodology: In order to understand the physiological mechanism behind the plant defences in Musa during the interaction with P. fijiensis, a transcriptomic and metabolomic analysis were led with two Musa genotypes differing in their resistance to the BSD development: The susceptible genotype “Williams” and the resistant genotype “Calcutta 4”. RNA-seq with Illumina technology, Nuclear Magnetic Resonance and Mass Spectrometry were the analytical techniques used for sample analysis.
Findings: The results clearly showed a fast and early plant response in the resistant genotype “Calcutta 4”, mainly with the induction of a group of genes and metabolites involved in pathogen recognition, hormonal signal transduction and pathogenesis-related proteins whereas a poor induction of those physiological responses were detected in the susceptible genotype “Williams”.
Conclusion & Significance: Our results support new insights about the role of JA-Et signalling pathways play in the response of
the resistant banana genotype Calcutta 4 during the pathogen attack. Furthermore, it opens the possibility to explore whether
these defence mechanism reported here are similar for other resistant Musa genotypes.
Philip J Jackson
University of Sheffield, UK
Title: Proteomic adaptation in high and low light ecotypes of the marine picocyanobacterium Prochlorococcus marinus, an important global primary producer
Time : 13:05-13:35
Biography:
Philip J Jackson obtained his PhD in Biochemistry from the University of Leeds, UK where his research is centered on the control of ATP synthesis in mitochondria.His subsequent postdoctoral positions, also in Leeds, involved the characterization of potential glycoprotein tumour biomarkers and the structural analysis of several membrane-intrinsic proteins including a proton-translocating ATPase. He then joined an instrumentation manufacturer as a product application specialist in proteomics and lipidomics. In 2010, he returned to Postdoctoral research to work in the Laboratories of Professors Neil Hunter, FRS and Mark Dickman at the University of Sheffield (UK). He specializes in biological mass spectrometry, applying this technology to (1) quantitative proteomics of complexes involved in light harvesting and energy transfer in photosynthesis, (2) analysis of protein-protein interactions within complexes that direct photosystem assembly and (3) structural characterization of the enzyme systems responsible for chlorophyll biosynthesis.
Abstract:
Prochlorococcus marinus, an oceanic phototrophic picocyanobacterium, is probably the most abundant organism on earth with an estimated population of 2.9±0.1 x 1027 cells. Consequently it is an important global primary producer responsible for the fixation of 4.0 x 106 tonnes of carbon per annum. In terms of habitat, Prochlorococcus has been recovered from diverse locations at depths down to 200 m. Given the wide distribution of Prochlorococcus within the euphotic zone, genetically distinct ecotypes have evolved in response to the level of sunlight penetration and nutrient availability. The four examples used for this study were: (1) MED4 (Mediterranean 5 m), (2) NATL2A (N. Atlantic 10 m), (3) SS120 (Sargasso 120 m) and (4) MIT9313 (Gulf Stream 135 m). Cells were grown under very low intensity illumination, similar to that experienced by the deep water ecotypes. After isolating thylakoid membranes, proteins were extracted, digested with endoproteinase Lys-C and trypsin, then analyzed by nanoLC-MS/MS. Identified proteins were quantified by the label-free iBAQ method which was validated by the expected PSI:PSII ratio of 3-4 for Synechocystis as determined by spectroscopy. The Prochlorococcus ecotypes showed markedly different PSI:PSII ratios to Synechocystis: near to 1:1 in MED4 and MIT9313, and 0.5 in NATL2A and SS120. Therefore there appears not to be a simple relationship between PSI:PSII ratio and illumination. On the other hand, amounts of high light inducible proteins (HLIPs), associated with photoprotection, and the relative amounts of light dependent and independent POR enzymes, occurring in the chlorophyll biosynthesis pathway did reflect the expected habitat illumination levels in the Prochlorococcus ecotypes and Synechocystis. As the Prochlorococcus ecotypes were grown under the same low light intensity, the expression patterns observed in this study appear to be an inherent feature of ecotypic adaptation to light intensity within their specific habitats.
Break: Lunch Break 13:35-14:35 @ La Place AB
Masahiro Onuma
Trisguide CO.,LTD, Japan
Title: Electronic water can reduce oxidative stress in cancer and diabetes patients for 3 weeks drinking
Time : 14:35-15:05
Biography:
Masahiro Onuma pursued his Bachelors in Biochemistry from Tokyo University of Agriculture and Technology. He is the President of TriSGuide Ltd. He has expertise in oxidative disease prevention to use non-medical product based on GSK’s experience of allopurinol which is the strongest anti-oxidant in this world.
Abstract:
Oxidative stress means a state where there is an imbalance between the oxidizing action and the reducing action due to reactive oxygen species (ROS) in a living body, resulting in the oxidizing action becoming dominant. Oxidative stress arises as the balance between production and removal is disrupted through excessive production of ROS and impairment of the antioxidant system. Oxidative stress has been reported to be involved in the onset and progress of various diseases. Characteristics of Type 2 diabetes are insulin secretion failure and insulin resistance, but it seems that oxidative stress is greatly involved in insulin secretion failure. In the insulin secretion-inducing β cells of Langerhans islets in the pancreas, the amount of superoxide dismutase (SOD), which is representative of the ROS elimination system, is small and resistance to oxidative stress is considered to be weak. Regarding cancer, it is well known that chronic inflammatory conditions increase the risk of carcinogenesis. Cells
such as neutrophils and macrophages are activated in the inflammation area leading to increase in production of active oxygen and nitric oxide. These free radicals cause DNA mutation and cell proliferation thereby promoting cancer development. When chronic inflammation is present, cancer develops more easily. Electronic water, which was developed to generate electron in water, was consumed for three weeks, after meals, between meals and before sleeping 6 times a day, and according to the test subjects' possible time periods. The amount of drinking water was 750-1000 mL, and BAP and d-ROMs checks for all cases were carried out at 4:30 pm. The results of cancer patients and diabetes patients were seen as attached. As a result, the d-ROMs value in the degree of oxidative stress has reduced, and the BAP value, which is an indicator of plasma antioxidant capacity, has improved
significantly.
Halina Malina
Axanton Technology GmbH, Switzerland
Title: Elimination the chemically modified proteins from the cell membranes prevents the cell degeneration
Time : 15:05-15:35
Biography:
Halina Malina pursued her PhD in antibiotic biosynthesis working at the Medical School of Lodz (Poland). She is a Chemical Engineer from Polytechnic of Lodz, Lodz. Since 1984, she worked at the Institute of Chemistry of Natural Substances (CNRS), with collaboration with Pasteur Institute of Paris (France). In 1990 she worked as project leader in CNRS University in Zurich. She discovered the IDO in the eye and the role of xanthurenic acid in the cataract development. She followed the research on xanthurenic acid-induced cell pathology at universities in Lausanne, and Bern and ETH. Her research on the chemical mechanism of the diseases with aging met a strong opposition of the academia healing the transgenic mice. All support for research was rejected in 2004. She continued on her own and established the technology curing the infection and the aging-associated pathology.
Abstract:
Statement of the Problem: Most young people are healthy, and with aging, they develop the degenerative diseases. In the last thirty years, it was thought that the disorders in aging are genetic and cannot be prevented. We present here the proof of concept that the aging-associated diseases are not genetic, but develops because of the chemically modified proteins. The discovery leads to effective prevention of the cell degeneration in aging and infection permitting to keep people healthy longer and improving the quality of life.
Methodology & Theoretical Orientation: The peptides corresponding to the intrinsically disordered sequences were synthesized in vitro and incubated with xanthurenic acid to obtain the covalently modified polymers observed on the SDSPAGE (patents). The polymers were injected into rabbits. The IgG's were isolated and tested in the primary cell cultures, in vivo in mice (outsourcing) and human, directly used by the investigator for her dermatitis and the metabolic disorder.
Findings: In a primary cell culture Xan led to the cell death caused by the covalently modified proteins. The regulatory sequences of the proteins, called intrinsically disordered sequences (IDSeqs) are preferentially modified. Any cell system cannot remove the chemically modified and polymerized IDSeqs. They alter the cell membranes leading to the caspases activation and cell degeneration.The author established a new technology targeting the chemically modified proteins. In mice, the molecule establishing membranes (MEMS) healed the Staphylococcus aureus and restored immunity in cyclophosphamide-treated mice using MEMS A-144 at five micrograms, weekly, on the mouth mucosa cured skin infection and cardiovascular disorder.
Conclusion & Significance: The chemically modified proteins must be eliminated to restore the cell homeostasis. MEMS stops the upstream cause of the pathology in infection or aging.
Vivek Kamath
Heal The World, India
Title: Case study of insulin dependent diabetes: Healing without medicines
Time : 15:35-16:05
Biography:
Vivek Kamath is the founder of heal the world organization is a Reiki Master, Mexican Healer, Melchizedek Healer, Crystal Healer, and Past Life Regression Therapy Expert. He has healed many diabetic patients (Type1, Type2, Type 3/1.5/LADA) without any medicines. He has also healed Cancer (stage 4) blood pressure (both high and low blood pressure), heart disease (removed the heart blockages), removed kidney stones, ovarian cysts, fibrosis of the breast, fatty liver, lungs disease, cured sinusitis, severe joint pain, lumbar L5 spinal disk pain, Sciatica pain, neck pain, constipation, rheumatoid arthritis, glaucoma, migraines, headaches, insomnia, stomach related problem, IBS, diabetic gum problems, skin problems( dry skin, eczema) and chronic nasal allergies, nasal blockages without any medicines. Some of the above treatments have been completed within a week to maximum 1-month duration.
Abstract:
As most of us are aware Type 2 diabetes can be controlled and cured completely with the diet, workouts (yoga), effective stress management and other healing methods. Testimonial of one of my patient aged 70 years, male suffering from diabetes from more than 12 years and took 3 insulin doses per day. He has completed Reiki Master Degree through some Reiki Master in India. He came to be with the problem that he is not able to feel the energy while doing his self healing. Moreover, he was suffering major health problems like (Insulin dependent diabetes for nearly 12 years, insomnia, constipation, stomach heaviness, etc.). I gave him Reiki Level 1 attunement a month back. On 25th July he has taken his random glucose reading. The random glucose level was 575 mg/dl. On July 27th I have given him Usui Reiki Level 2, 3 Attunement, Karuna Reiki Attunement and Mexican Healing attunement (Guru Poornima Day). Immediately after the attunement we checked his random glucose level it came down to 383 mg/Dl. After 1 Mexican Healing: On 28th July evening, I conducted Mexican healing training class to my patient along with others. I took his random glucose level before giving him Mexican healing and it was 344 mg/Dl. After 1 Mexican healing his glucose came down to 260 mg/Dl. All the people who attended the training class were astonished with the power of Mexican healing. To summarize in 3 days, using attunement (Reiki and Mexican attunement) and with 1 Mexican healing, his glucose level came down from 575 mg/dL to 260 mg/DL. What is the percentage in drop down? 54.8%. Is possible to bring this much of level down in 3 days without taking insulin? Please note that there is no change in his food/diet or workouts. Also note the difference between the readings after attunement and after healing. We are all made up of energy and it flows though the energy centers (chakra). We can’t necessarily see energy, but we can feel the energy. When our energy centers becomes weak, diseases related to corresponding energy centers will pop up. For Diabetes, there is energy center called “Solar Plexus” (Also called as Manipura Chakra). All diabetic patients will have low energy in their solar plexus energy. If you energize this energy center you will see the drastic reduction in your glucose level. You do not need to go for a pancreas transplant or tablets/insulin to treat your diabetes. It is all in the energy. Learn from your Reiki master or healers to energize all your weaker organs or energy centers.
Break: Networking & Refreshments 16:05-16:25 @ Europa Foyer