This document summarizes an investigation into how gut microflora affects endogenous metabolic pathways using mass spectrometry-based metabolomics. Pseudo germ-free rats were used as a model by administering antibiotics to suppress gut microflora. Urine metabolites were then analyzed and compared between control and pseudo germ-free rats using UPLC-QTOF-MS. Validation steps included repeatability testing of QC samples and a test mixture to ensure method reliability. Multivariate analysis of metabolic profiles identified differences between control and pseudo germ-free groups, suggesting gut microflora influences certain metabolic pathways.
This document discusses the evaluation of drug toxicity through metabolomic profiling. It describes how metabolomics can be used to detect liver and kidney toxicity induced by various compounds. The study designs involve administering hepatotoxic or nephrotoxic drugs to animals and collecting blood, urine, and tissue samples for analysis. Nuclear magnetic resonance spectroscopy is used to analyze metabolic profiles in biofluids and multivariate statistical analysis is applied to identify biomarkers of toxicity. The document outlines several studies conducted by the National Institute of Food and Drug Safety Evaluation investigating drug toxicity using metabolomics approaches.
Extraction And Characterization Of Protease From The Viscera Of Skipjack Tuna...jeni_anggrek10
The document summarizes research on extracting and characterizing protease from the viscera of skipjack tuna fish. Protease was extracted from the intestines, stomach, pancreas, and liver of skipjack tuna using potassium phosphate solution and precipitated using either cold acetone or ammonium sulfate. The protease showed highest activity when precipitated with a 1:2 ratio of extract to acetone. The optimal temperature and pH for the acetone-precipitated protease were 50°C and 8, respectively. Tests on temperature and pH stability as well as effects of additives like NaCl, CaCl2, and EDTA were also conducted to characterize the protease.
This document summarizes a study that used SWATH-based mass spectrometry to compare the proteomes of the BCG-Korea strain and BCG-Pasteur strain of Mycobacterium bovis. Twenty of the most abundant proteins identified in the BCG proteome were selected for quantification between the two strains using SWATH. Thirteen of the twenty proteins showed significant changes in expression levels between the BCG-Korea and BCG-Pasteur strains, indicating that genomic differences between the strains affect protein expression levels. The SWATH method provided a straightforward, reliable approach for comparative proteomic analysis of the two BCG strains.
IOSR Journal of Pharmacy (IOSRPHR), www.iosrphr.org, call for paper, research...iosrphr_editor
This document summarizes a study that tested the effects of different concentrations of anti-glucosyltransferase (anti-GTF) antibody on the growth of Streptococcus sobrinus bacteria and the enzymatic activity of purified GTF-Ib enzyme isolated from the bacteria. The study found that various concentrations of inhibitors like sodium fluoride, chlorohexidine, and ZAK mouthrinse, as well as the anti-GTF antibody, were able to inhibit the enzymatic activity of GTF-Ib, with the exception of EDTA. Specifically, the anti-GTF antibody at a concentration of 1.5x10-3 mM inhibited 87.33% of GTF-Ib enzymatic
This document describes analyzing the minor element composition of yeast extract using quantitative spectrochemical methods. The results showed yeast extract contains various minor elements including aluminum, barium, copper, iron, magnesium, manganese, strontium, cadmium, cobalt, chromium, gallium, molybdenum, nickel, lead, tin, titanium, vanadium, and zinc. Providing this detailed elemental analysis of yeast extract facilitates identifying which specific elements are required for growth of microorganisms in chemically defined media.
This document summarizes the development of a new high-throughput method for single nucleotide polymorphism (SNP) genotyping using flow cytometric analysis of fluorescent microspheres. Key points:
- A multiplexed assay was developed that uses PCR-amplified DNA containing the SNP of interest, a capture probe with a unique "ZipCode" sequence, and fluorescent dyes to label extension products on the microspheres for analysis by flow cytometry.
- The assay was tested on 20 SNPs from 633 patients, showing 99.3% agreement with established gel-based genotyping methods. Signal was improved 5-10x using an indirect fluorescent labeling method.
- A new allele-specific primer extension assay was
NOAEL Cancer Therapy is a novel anti-cancer therapy to overcome conventional chemotherapy by controlling toxicity, based on restless & repeated ‘below-NOAEL‘ doses of Polytaxel for complete regression of tumor without severe adverse effects Polytaxel: the first ‘pain-free’ anti-cancer drug developed for NOAEL Cancer Therapy, based on the platform technology of polyphosphazene conjugation
NOAEL Cancer Therapy’s first xenograft test result on ‘hard-to-treat’ pancreatic cancer model indicates possibility that pancreatic cancer can be completely treated with below-NOAEL doses
This document discusses the evaluation of drug toxicity through metabolomic profiling. It describes how metabolomics can be used to detect liver and kidney toxicity induced by various compounds. The study designs involve administering hepatotoxic or nephrotoxic drugs to animals and collecting blood, urine, and tissue samples for analysis. Nuclear magnetic resonance spectroscopy is used to analyze metabolic profiles in biofluids and multivariate statistical analysis is applied to identify biomarkers of toxicity. The document outlines several studies conducted by the National Institute of Food and Drug Safety Evaluation investigating drug toxicity using metabolomics approaches.
Extraction And Characterization Of Protease From The Viscera Of Skipjack Tuna...jeni_anggrek10
The document summarizes research on extracting and characterizing protease from the viscera of skipjack tuna fish. Protease was extracted from the intestines, stomach, pancreas, and liver of skipjack tuna using potassium phosphate solution and precipitated using either cold acetone or ammonium sulfate. The protease showed highest activity when precipitated with a 1:2 ratio of extract to acetone. The optimal temperature and pH for the acetone-precipitated protease were 50°C and 8, respectively. Tests on temperature and pH stability as well as effects of additives like NaCl, CaCl2, and EDTA were also conducted to characterize the protease.
This document summarizes a study that used SWATH-based mass spectrometry to compare the proteomes of the BCG-Korea strain and BCG-Pasteur strain of Mycobacterium bovis. Twenty of the most abundant proteins identified in the BCG proteome were selected for quantification between the two strains using SWATH. Thirteen of the twenty proteins showed significant changes in expression levels between the BCG-Korea and BCG-Pasteur strains, indicating that genomic differences between the strains affect protein expression levels. The SWATH method provided a straightforward, reliable approach for comparative proteomic analysis of the two BCG strains.
IOSR Journal of Pharmacy (IOSRPHR), www.iosrphr.org, call for paper, research...iosrphr_editor
This document summarizes a study that tested the effects of different concentrations of anti-glucosyltransferase (anti-GTF) antibody on the growth of Streptococcus sobrinus bacteria and the enzymatic activity of purified GTF-Ib enzyme isolated from the bacteria. The study found that various concentrations of inhibitors like sodium fluoride, chlorohexidine, and ZAK mouthrinse, as well as the anti-GTF antibody, were able to inhibit the enzymatic activity of GTF-Ib, with the exception of EDTA. Specifically, the anti-GTF antibody at a concentration of 1.5x10-3 mM inhibited 87.33% of GTF-Ib enzymatic
This document describes analyzing the minor element composition of yeast extract using quantitative spectrochemical methods. The results showed yeast extract contains various minor elements including aluminum, barium, copper, iron, magnesium, manganese, strontium, cadmium, cobalt, chromium, gallium, molybdenum, nickel, lead, tin, titanium, vanadium, and zinc. Providing this detailed elemental analysis of yeast extract facilitates identifying which specific elements are required for growth of microorganisms in chemically defined media.
This document summarizes the development of a new high-throughput method for single nucleotide polymorphism (SNP) genotyping using flow cytometric analysis of fluorescent microspheres. Key points:
- A multiplexed assay was developed that uses PCR-amplified DNA containing the SNP of interest, a capture probe with a unique "ZipCode" sequence, and fluorescent dyes to label extension products on the microspheres for analysis by flow cytometry.
- The assay was tested on 20 SNPs from 633 patients, showing 99.3% agreement with established gel-based genotyping methods. Signal was improved 5-10x using an indirect fluorescent labeling method.
- A new allele-specific primer extension assay was
NOAEL Cancer Therapy is a novel anti-cancer therapy to overcome conventional chemotherapy by controlling toxicity, based on restless & repeated ‘below-NOAEL‘ doses of Polytaxel for complete regression of tumor without severe adverse effects Polytaxel: the first ‘pain-free’ anti-cancer drug developed for NOAEL Cancer Therapy, based on the platform technology of polyphosphazene conjugation
NOAEL Cancer Therapy’s first xenograft test result on ‘hard-to-treat’ pancreatic cancer model indicates possibility that pancreatic cancer can be completely treated with below-NOAEL doses
Publication - Alternative Surfactants for Improved Efficiency of In Situ Tryp...Nathan Marshall
This document summarizes a study investigating the use of alternative surfactants to improve the efficiency of in situ tryptic proteolysis of fingermarks. The study tested a range of non-ionic and anionic surfactants individually and in combination, including MEGA-8, OcGlu, OcThio, DDM, and RapiGest SF. Data demonstrated that higher percentages of MEGA-8 as well as combinations of detergents resulted in more peptide peaks detected from fingermarks. RapiGest SF, normally used for solution digestions, was also shown to improve in situ proteolysis. Similar results were observed on rat brain tissue sections. Overall, the study indicates that surfactants can enhance the efficiency
The document summarizes an in vitro high content screening platform for assessing drug-induced hepatotoxicity through multiple mechanisms. The platform utilizes HepG2 human hepatocellular carcinoma cells in assays to measure cell proliferation, apoptosis, phospholipidosis, steatosis, cell cycle effects, and inflammatory cytokine secretion. Assays are performed in a multiplexed format using automated fluorescent microscopy and xMAP technology. The platform provides quantitative data on toxicity mechanisms and allows ranking of compounds based on their hepatotoxic potential to help guide drug development.
Nutrition Reseach - Enhanced brown adipose tissue differentiation in PTP1B kn...Aroldo Trejo
- The study investigated the effects of knocking out PTP1B on brown adipose tissue (BAT) differentiation in rats. Cells from PTP1B knockout rats were treated with human PTP1B that was either wild type (WT), knockout (KO), or catalytically impaired (D/A).
- After 8 days of differentiation, Oil Red O staining showed that the KO and D/A treatments displayed greater differentiation (152% and 154% respectively) compared to WT (136%), although the differences were not statistically significant.
- Western blot analysis confirmed presence of PTP1B in WT and D/A cells but not in KO cells, supporting the role of PTP1B in inhibiting
Evgeny nikolaev proteomics of body liquids as a source for potential methods ...igorod
The document discusses using mass spectrometry to analyze body fluids like urine, saliva, and exhaled breath condensate for medical diagnostics and biomarker discovery. It describes creating databases of accurate mass tags and retention times from mass spec analyses of peptides and proteins in body fluids to enable fast identification. Biomarkers found for diseases like COPD, pneumonia and changes after lung transplantation are mentioned.
This document discusses the potential for in vivo imaging as a powerful alternative to traditional animal studies in ecotoxicology. It notes that in vivo studies are required under REACH regulations to evaluate chemical risks but have ethical and economic drawbacks due to their use of large animal numbers. In vivo imaging can help address these issues by providing quantitative, precise, and predictive data while reducing experimental artifacts and animal usage through longitudinal studies. The document then provides several examples of how different in vivo imaging modalities such as MRI, optical imaging, PET, CT, and ultrasound have been applied to study inflammation, cell metabolism, bone development, and embryo toxicity in ecotoxicology. It argues that in vivo imaging can help achieve the goals of the 3Rs and
This document summarizes research into inhibiting the virulence of gram-positive bacteria by inhibiting the enzyme sortase A. The researchers used protein docking software to predict small molecule inhibitors from a library of compounds synthesized via the Ugi 4-component reaction. Solubility modeling was used to select Ugi products likely to precipitate out of solution and be purified easily. Selected inhibitors were tested in a colorimetric assay to measure their ability to inhibit sortase A and decrease bacterial virulence. Future work will optimize reactions, docking, and assays to identify potent synthetic sortase A inhibitors.
Dr. Jesús Jiménez Barbero - Simposio Internacional 'Química: respuestas para ...Fundación Ramón Areces
Los días 8 y 9 de octubre de 2014, la Fundación Ramón Areces acogió el Simposio Internacional 'Química: respuestas para una vida mejor', organizado en colaboración con la Fundación General CSIC. Su finalidad fue ofrecer a los participantes una visión atractiva de la química moderna que sirva de base al desarrollo de nuevas respuestas para una sociedad en rápida evolución.
Effects of extract of Drymaria cordata on isolated rat liver MMPT poreGloria Okenze
This study investigated the effects of extract of Drymaria cordata, a medicinal plant, on the mitochondrial membrane permeability transition pore (MMPT pore) using isolated rat liver mitochondria. The objectives were to determine if the extract could induce opening of the MMPT pore. Fractions of the methanol extract were tested for their effects on mitochondrial swelling, ATPase activity, and lipid peroxidation in the presence and absence of calcium. The chloroform fraction was found to be the most potent at inducing MMPT pore opening and inhibiting lipid peroxidation. This suggests the chloroform fraction contains compounds that could modulate apoptosis by affecting the mitochondrial membrane and MMPT pore. Further studies are needed to identify active compounds and fully characterize
dual-event machine learning models to accelerate drug discoverySean Ekins
This document describes using dual-event machine learning models to accelerate drug discovery for tuberculosis (TB). Bayesian machine learning models were generated using molecular fingerprints and descriptors from large TB screening datasets. The models were able to retrospectively and prospectively predict active compounds from both internal and external datasets with good enrichment. A prospective virtual screen of an antimalarial library identified a hit with good efficacy and selectivity in vitro and in vivo. Overall, the dual-event models integrating phenotypic screening data with cytotoxicity were able to identify several novel TB drug leads and represent an approach to accelerate drug discovery.
General conclusions
- Current methods used by the industry to evaluate protein quality are not capable of detecting existing differences among SBM
- The composition and the protein quality of SBM vary with the origin of the bean
- Different matrixes should be used for SBM of different origins, NIR technology might help
- Proteases might improve the uniformity and nutritive value of SBM batches
This study aimed to develop a non-destructive method of measuring lactate concentration in atherosclerotic plaques using near-infrared spectroscopy. Human carotid plaques were maintained ex vivo under physiologic conditions for extended measurements. Measurements of pH, temperature, PO2 and PCO2 over time validated plaque viability. Near-infrared spectroscopy measurements of lactate correlated well with destructive assays, suggesting feasibility of non-destructively assessing plaque activity and vulnerability.
This study aimed to develop a non-destructive method of measuring lactate concentration in atherosclerotic plaques using near-infrared spectroscopy. Human carotid plaques were maintained ex vivo under physiologic conditions for extended measurements. Measurements of pH, temperature, PO2 and PCO2 over time validated plaque viability. Near-infrared spectroscopy measurements of lactate correlated well with destructive assays, suggesting feasibility of non-destructively assessing plaque activity and vulnerability.
The document describes progress made in calibrating a 3 French near infrared spectroscopy fiber optic catheter to non-invasively measure pH and lactate levels in atherosclerotic plaque. Researchers were able to maintain excised human carotid plaques under near physiological conditions for extended periods to collect measurements. Preliminary results show the near infrared measurement of tissue lactate in living plaques correlates well with destructive standard measurements, and this approach may allow direct translation of calibration equations to future in vivo validation studies.
Focus on aggregation: types, causes, characterization, and impactKBI Biopharma
This document discusses protein aggregation, including types and sizes of aggregates, mechanisms of aggregation, and challenges in characterization. It summarizes sedimentation velocity and field-flow fractionation as useful analytical methods to cross-check size exclusion chromatography for characterizing aggregates. Sedimentation velocity can resolve multiple aggregate peaks but requires expertise, while field-flow fractionation has less surface interaction but high method development. No single method detects all aggregate types.
This document summarizes a study that investigated the effects of zinc oxide nanoparticles on the differentiation of human mesenchymal stem cells into osteoblasts. The study exposed stem cells to 30 and 60 μg/ml of 30 nm zinc oxide nanoparticles and found that exposure to 30 μg/ml significantly increased the expression of osteogenic marker genes and mineralization compared to the control and 60 μg/ml exposure group. The results indicate that zinc oxide nanoparticles can enhance stem cell differentiation into osteoblasts in a dose-dependent manner.
Tracel 2011 3rd international symposium on trace elements & healthOBenavente
This poster presents research on the potential of carnosic acid and other compounds as radiation countermeasures. Researchers evaluated the genoprotective effects of carnosic acid and other antioxidants against damage from ionizing radiation using the cytokinesis-block micronucleus test on human lymphocytes before and after gamma radiation exposure. Carnosic acid and other compounds showed significant genoprotection, reducing the frequency of micronuclei by up to 50%. Researchers also found that carnosic acid increased cell survival in prostate and melanoma cell lines when administered before X-ray exposure, indicating radioprotective effects. The results suggest carnosic acid may help protect against the damaging effects of radiation exposure and warrants further study as
Objective(s):
This study aimed to address the gold nanoparticle(GNP)-dose and exposure duration effect on the kidney function of rats: in vivo.
Materials and Methods:
A total of 32 healthy male Wistar rats were used in this study. Animals were randomly divided into groups, three GNP-treated groups and control group. Group 1, 2 and 3 received. /5 cc of solution containing 5, 10,100 ppm Au via IP injection for 7 successive days, respectively. The control group was treated with 0.5% normal saline. Several biochemical parameters such as BUN (blood urea nitrogen), creatine and uric acid were evaluated at various time points (7 and 14 days). After 14 days, the tissue of kidney was collected and investigated.
Results:
There was no significant difference between the control and the intervention group regarding the amount of creatine-BUN and uric acid. The amount of creatine-BUN and uric acid showed increase in all the groups [except group1 (creatine) and group 2 (uric acid)] in the 7 and 14 days after intervention compared to the control group, but this difference was not significant. Results of histopatological tissue kidney showed: in group 1 and 3, complete destruction of the proximal tubules and distal cortical, in group 2, almost complete destruction of proximal tubules and distal.
Conclusions:
The induced histological alterations might be an indication of injured renal tubules due to GNPs toxicity that become unable to deal with the accumulated residues resulting from metabolic and structural disturbances caused by these NPs
This document discusses using metabolomics to analyze the food metabolome - all metabolites derived from digestion and metabolism of food components. It notes the complexity of nutritional exposures from foods, which contain nutrients, non-nutrients from natural and man-made sources, and contaminants. Individual metabolic capacity is also influenced by genetics, microbiota, age, and other factors. The food metabolome is defined and applications are described, including discovering new food bioactives, understanding diet-health relationships, and personalized nutrition. Challenges in analyzing the large and diverse food metabolome are also outlined. Initial studies using targeted and untargeted metabolomics are discovering new biomarkers of food intake from various foods. Larger cohort studies and controlled interventions are needed
The document discusses the human gut and how it has evolved dynamically over time. It highlights several key factors that have influenced gut evolution, including shifts to exogenous food processing, the production and role of gut microbes, and cultural practices around food preparation. The gut microbiomes of other primates are only partially relevant to understanding human health, as the human gut and diet have undergone significant changes over the past 40,000 years through cultural innovations.
Metabolism is essential for eliminating drugs and other foreign substances from the body. Phase I metabolism makes compounds more polar through reactions like oxidation and hydrolysis. Phase II metabolism further increases polarity through conjugation. The liver is the primary site of metabolism, with cytochrome P450 enzymes performing many Phase I reactions. Metabolism inactivates most compounds but can also activate prodrugs. Factors like induction, inhibition, disease, age, sex and genetics can impact an individual's drug metabolism. Proper metabolism and excretion of compounds and their metabolites terminates their biological activity.
Publication - Alternative Surfactants for Improved Efficiency of In Situ Tryp...Nathan Marshall
This document summarizes a study investigating the use of alternative surfactants to improve the efficiency of in situ tryptic proteolysis of fingermarks. The study tested a range of non-ionic and anionic surfactants individually and in combination, including MEGA-8, OcGlu, OcThio, DDM, and RapiGest SF. Data demonstrated that higher percentages of MEGA-8 as well as combinations of detergents resulted in more peptide peaks detected from fingermarks. RapiGest SF, normally used for solution digestions, was also shown to improve in situ proteolysis. Similar results were observed on rat brain tissue sections. Overall, the study indicates that surfactants can enhance the efficiency
The document summarizes an in vitro high content screening platform for assessing drug-induced hepatotoxicity through multiple mechanisms. The platform utilizes HepG2 human hepatocellular carcinoma cells in assays to measure cell proliferation, apoptosis, phospholipidosis, steatosis, cell cycle effects, and inflammatory cytokine secretion. Assays are performed in a multiplexed format using automated fluorescent microscopy and xMAP technology. The platform provides quantitative data on toxicity mechanisms and allows ranking of compounds based on their hepatotoxic potential to help guide drug development.
Nutrition Reseach - Enhanced brown adipose tissue differentiation in PTP1B kn...Aroldo Trejo
- The study investigated the effects of knocking out PTP1B on brown adipose tissue (BAT) differentiation in rats. Cells from PTP1B knockout rats were treated with human PTP1B that was either wild type (WT), knockout (KO), or catalytically impaired (D/A).
- After 8 days of differentiation, Oil Red O staining showed that the KO and D/A treatments displayed greater differentiation (152% and 154% respectively) compared to WT (136%), although the differences were not statistically significant.
- Western blot analysis confirmed presence of PTP1B in WT and D/A cells but not in KO cells, supporting the role of PTP1B in inhibiting
Evgeny nikolaev proteomics of body liquids as a source for potential methods ...igorod
The document discusses using mass spectrometry to analyze body fluids like urine, saliva, and exhaled breath condensate for medical diagnostics and biomarker discovery. It describes creating databases of accurate mass tags and retention times from mass spec analyses of peptides and proteins in body fluids to enable fast identification. Biomarkers found for diseases like COPD, pneumonia and changes after lung transplantation are mentioned.
This document discusses the potential for in vivo imaging as a powerful alternative to traditional animal studies in ecotoxicology. It notes that in vivo studies are required under REACH regulations to evaluate chemical risks but have ethical and economic drawbacks due to their use of large animal numbers. In vivo imaging can help address these issues by providing quantitative, precise, and predictive data while reducing experimental artifacts and animal usage through longitudinal studies. The document then provides several examples of how different in vivo imaging modalities such as MRI, optical imaging, PET, CT, and ultrasound have been applied to study inflammation, cell metabolism, bone development, and embryo toxicity in ecotoxicology. It argues that in vivo imaging can help achieve the goals of the 3Rs and
This document summarizes research into inhibiting the virulence of gram-positive bacteria by inhibiting the enzyme sortase A. The researchers used protein docking software to predict small molecule inhibitors from a library of compounds synthesized via the Ugi 4-component reaction. Solubility modeling was used to select Ugi products likely to precipitate out of solution and be purified easily. Selected inhibitors were tested in a colorimetric assay to measure their ability to inhibit sortase A and decrease bacterial virulence. Future work will optimize reactions, docking, and assays to identify potent synthetic sortase A inhibitors.
Dr. Jesús Jiménez Barbero - Simposio Internacional 'Química: respuestas para ...Fundación Ramón Areces
Los días 8 y 9 de octubre de 2014, la Fundación Ramón Areces acogió el Simposio Internacional 'Química: respuestas para una vida mejor', organizado en colaboración con la Fundación General CSIC. Su finalidad fue ofrecer a los participantes una visión atractiva de la química moderna que sirva de base al desarrollo de nuevas respuestas para una sociedad en rápida evolución.
Effects of extract of Drymaria cordata on isolated rat liver MMPT poreGloria Okenze
This study investigated the effects of extract of Drymaria cordata, a medicinal plant, on the mitochondrial membrane permeability transition pore (MMPT pore) using isolated rat liver mitochondria. The objectives were to determine if the extract could induce opening of the MMPT pore. Fractions of the methanol extract were tested for their effects on mitochondrial swelling, ATPase activity, and lipid peroxidation in the presence and absence of calcium. The chloroform fraction was found to be the most potent at inducing MMPT pore opening and inhibiting lipid peroxidation. This suggests the chloroform fraction contains compounds that could modulate apoptosis by affecting the mitochondrial membrane and MMPT pore. Further studies are needed to identify active compounds and fully characterize
dual-event machine learning models to accelerate drug discoverySean Ekins
This document describes using dual-event machine learning models to accelerate drug discovery for tuberculosis (TB). Bayesian machine learning models were generated using molecular fingerprints and descriptors from large TB screening datasets. The models were able to retrospectively and prospectively predict active compounds from both internal and external datasets with good enrichment. A prospective virtual screen of an antimalarial library identified a hit with good efficacy and selectivity in vitro and in vivo. Overall, the dual-event models integrating phenotypic screening data with cytotoxicity were able to identify several novel TB drug leads and represent an approach to accelerate drug discovery.
General conclusions
- Current methods used by the industry to evaluate protein quality are not capable of detecting existing differences among SBM
- The composition and the protein quality of SBM vary with the origin of the bean
- Different matrixes should be used for SBM of different origins, NIR technology might help
- Proteases might improve the uniformity and nutritive value of SBM batches
This study aimed to develop a non-destructive method of measuring lactate concentration in atherosclerotic plaques using near-infrared spectroscopy. Human carotid plaques were maintained ex vivo under physiologic conditions for extended measurements. Measurements of pH, temperature, PO2 and PCO2 over time validated plaque viability. Near-infrared spectroscopy measurements of lactate correlated well with destructive assays, suggesting feasibility of non-destructively assessing plaque activity and vulnerability.
This study aimed to develop a non-destructive method of measuring lactate concentration in atherosclerotic plaques using near-infrared spectroscopy. Human carotid plaques were maintained ex vivo under physiologic conditions for extended measurements. Measurements of pH, temperature, PO2 and PCO2 over time validated plaque viability. Near-infrared spectroscopy measurements of lactate correlated well with destructive assays, suggesting feasibility of non-destructively assessing plaque activity and vulnerability.
The document describes progress made in calibrating a 3 French near infrared spectroscopy fiber optic catheter to non-invasively measure pH and lactate levels in atherosclerotic plaque. Researchers were able to maintain excised human carotid plaques under near physiological conditions for extended periods to collect measurements. Preliminary results show the near infrared measurement of tissue lactate in living plaques correlates well with destructive standard measurements, and this approach may allow direct translation of calibration equations to future in vivo validation studies.
Focus on aggregation: types, causes, characterization, and impactKBI Biopharma
This document discusses protein aggregation, including types and sizes of aggregates, mechanisms of aggregation, and challenges in characterization. It summarizes sedimentation velocity and field-flow fractionation as useful analytical methods to cross-check size exclusion chromatography for characterizing aggregates. Sedimentation velocity can resolve multiple aggregate peaks but requires expertise, while field-flow fractionation has less surface interaction but high method development. No single method detects all aggregate types.
This document summarizes a study that investigated the effects of zinc oxide nanoparticles on the differentiation of human mesenchymal stem cells into osteoblasts. The study exposed stem cells to 30 and 60 μg/ml of 30 nm zinc oxide nanoparticles and found that exposure to 30 μg/ml significantly increased the expression of osteogenic marker genes and mineralization compared to the control and 60 μg/ml exposure group. The results indicate that zinc oxide nanoparticles can enhance stem cell differentiation into osteoblasts in a dose-dependent manner.
Tracel 2011 3rd international symposium on trace elements & healthOBenavente
This poster presents research on the potential of carnosic acid and other compounds as radiation countermeasures. Researchers evaluated the genoprotective effects of carnosic acid and other antioxidants against damage from ionizing radiation using the cytokinesis-block micronucleus test on human lymphocytes before and after gamma radiation exposure. Carnosic acid and other compounds showed significant genoprotection, reducing the frequency of micronuclei by up to 50%. Researchers also found that carnosic acid increased cell survival in prostate and melanoma cell lines when administered before X-ray exposure, indicating radioprotective effects. The results suggest carnosic acid may help protect against the damaging effects of radiation exposure and warrants further study as
Objective(s):
This study aimed to address the gold nanoparticle(GNP)-dose and exposure duration effect on the kidney function of rats: in vivo.
Materials and Methods:
A total of 32 healthy male Wistar rats were used in this study. Animals were randomly divided into groups, three GNP-treated groups and control group. Group 1, 2 and 3 received. /5 cc of solution containing 5, 10,100 ppm Au via IP injection for 7 successive days, respectively. The control group was treated with 0.5% normal saline. Several biochemical parameters such as BUN (blood urea nitrogen), creatine and uric acid were evaluated at various time points (7 and 14 days). After 14 days, the tissue of kidney was collected and investigated.
Results:
There was no significant difference between the control and the intervention group regarding the amount of creatine-BUN and uric acid. The amount of creatine-BUN and uric acid showed increase in all the groups [except group1 (creatine) and group 2 (uric acid)] in the 7 and 14 days after intervention compared to the control group, but this difference was not significant. Results of histopatological tissue kidney showed: in group 1 and 3, complete destruction of the proximal tubules and distal cortical, in group 2, almost complete destruction of proximal tubules and distal.
Conclusions:
The induced histological alterations might be an indication of injured renal tubules due to GNPs toxicity that become unable to deal with the accumulated residues resulting from metabolic and structural disturbances caused by these NPs
This document discusses using metabolomics to analyze the food metabolome - all metabolites derived from digestion and metabolism of food components. It notes the complexity of nutritional exposures from foods, which contain nutrients, non-nutrients from natural and man-made sources, and contaminants. Individual metabolic capacity is also influenced by genetics, microbiota, age, and other factors. The food metabolome is defined and applications are described, including discovering new food bioactives, understanding diet-health relationships, and personalized nutrition. Challenges in analyzing the large and diverse food metabolome are also outlined. Initial studies using targeted and untargeted metabolomics are discovering new biomarkers of food intake from various foods. Larger cohort studies and controlled interventions are needed
The document discusses the human gut and how it has evolved dynamically over time. It highlights several key factors that have influenced gut evolution, including shifts to exogenous food processing, the production and role of gut microbes, and cultural practices around food preparation. The gut microbiomes of other primates are only partially relevant to understanding human health, as the human gut and diet have undergone significant changes over the past 40,000 years through cultural innovations.
Metabolism is essential for eliminating drugs and other foreign substances from the body. Phase I metabolism makes compounds more polar through reactions like oxidation and hydrolysis. Phase II metabolism further increases polarity through conjugation. The liver is the primary site of metabolism, with cytochrome P450 enzymes performing many Phase I reactions. Metabolism inactivates most compounds but can also activate prodrugs. Factors like induction, inhibition, disease, age, sex and genetics can impact an individual's drug metabolism. Proper metabolism and excretion of compounds and their metabolites terminates their biological activity.
Metabolites have various functions, including fuel, structure, signaling, stimulatory and inhibitory effects on enzymes, catalytic activity of their own (usually as a cofactor to an enzyme), defense, and interactions with other organisms (e.g. pigments, odorants, and pheromones).
Metabolome refers to the complete set of chemical compounds involved in an organism's metabolism (such as metabolic intermediates, hormones and other signaling molecules, and secondary metabolites)
Metabolomics is the scientific study of chemical processes involving metabolites. Metabolomics is a relatively new member to the ‘-omics’ family of systems biology technologies.
The document discusses the gut microbiota and its importance. It notes that the gut contains trillions of microbes, weighing up to 5 lbs, with thousands of bacterial species. The major phyla include Bacteroidetes, Firmicutes, Actinobacteria, and Proteobacteria. The gut microbiota plays key roles in digesting nutrients, preventing pathogenic growth, and supporting metabolic and immune system functions. Dysbiosis of the gut microbiota is associated with obesity, stress, anxiety, depression, autism and other conditions. Future research on the gut microbiota is needed to better understand its development and impact on health and disease.
Metabolomics aims to quantify all metabolites in a cellular system. The challenges are chemical complexity and heterogeneity of metabolites, dynamic range of measurements, and throughput. Metabolites can be analyzed using spectroscopy and mass spectrometry coupled with gas or liquid chromatography. NMR provides information on metabolites directly from biofluids with little sample preparation. GC-MS and LC-MS are commonly used, with LC-MS measuring a broader range of primary and secondary metabolites. Data integration and identification of specific metabolites remain challenges.
This document summarizes research on ginsenosides and their metabolites. It notes that ginsenosides have poor oral absorption but are transformed by gut bacteria into active metabolites that show better absorption. One metabolite, M1, has been found to inhibit tumor cell proliferation, invasion, and adhesion in vitro. Studies in humans and rats have demonstrated M1's inhibitory effects on tumor metastasis after oral administration. The document also discusses the metabolism and pharmacokinetics of M1.
This study analyzed the stability and aggregation behavior of six types of silver nanoparticles (AgNPs) with different surface coatings in ultrapure water, cell culture medium, and cell culture medium supplemented with bovine serum albumin (BSA). Dynamic light scattering, electrophoretic light scattering, and transmission electron microscopy showed that AgNPs aggregated and formed micrometric clusters in cell culture medium due to its higher ionic strength. Addition of BSA prevented aggregation by forming a protein corona around the AgNPs. Zeta potential measurements also indicated BSA incorporation stabilized the AgNP surface charge. Therefore, BSA provides steric stabilization of AgNPs in biologically relevant conditions and more research is needed to understand nanoparticle behavior in real
This document summarizes Adriana Zardini Buzatto's background and research experience in lipidomics and metabolomics. It outlines her educational background, including undergraduate and graduate degrees from universities in Brazil and Canada. It also provides an overview of several of her current and past research projects applying lipidomics techniques to study conditions like cystic fibrosis, spinal cord injury, and Parkinson's disease. These projects involve analyzing small biological samples using nanoLC-MS for lipid profiling to better understand disease mechanisms and identify potential biomarkers.
This presentation contains recommendations and requirements for the design of bioanalytical testing used in comparibility studies for biosimilar drug development using rituximab as an example
Application of Physiologically-based Kinetic Models in Exposure ModelingIES / IAQM
This document summarizes a land condition symposium presentation on applying physiologically-based kinetic models in land contamination exposure modelling. The presentation introduced key terms like bioaccessibility and bioavailability, and described a study that used physiologically-based pharmacokinetic (PBPK) models to improve understanding of actual human exposure to contaminants like arsenic, cadmium, chromium, nickel, and lead in soil. Soil, produce, blood, and urine samples were collected and analyzed from allotment users. PBPK models were produced and evaluated against literature data. The models can estimate uptake fractions and relative
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1. Investigation on the endogenous metabolic
change by intestinal gut microflora using
mass spectrometry based metabolomics
한국과학기술연구원
생체분자기능연구 센터
정병화
Relative concentrations of bacteria at various locations
within the gut.
Arq Bras Endocrinol Metab. 2009
2. The “OMICS” Cascade
What can happen
What appears to be happening
What makes it happen
What has happened and is happening
(the most predictive of phenotype)
Phenotype
3. Metabolite
(molecular weight:<1000Da)
Because
1) they are the end products of cellular
regulatory processes.
2) their levels can be regarded as the ultimate
response of biological systems to genetic or
environmental changes.
4. Class Genomics Proteomics Metabolomics
Subject Gene Protein Metabolite
mw of
>100,000 5,000 – 200,000 100 – 1,000
subject
Systemic DNA 2D-gel Hyphenated tech.
Technique peptide mass
sequencing fingerprinting NMR, MASS
Separation of protein Idetification and
Research
Gene mapping Identification of Quantitation
Area
protein function of metabolite
5. Major technologies for Metabolomics
MS and NMR:
MS NMR
Identify and quantify metabolite Does not rely on the separation
after separation by GC, HPLC, CE of the analytes
so on (all kind of metabolite can be
measured simultaneously)
Very sensitive and selective Has significant limitations of
sensitivity ( 10 mg/compound on
column)
Identification of metabolite
according to its fragmentation
pattern
Wikipedia
7. Gut is one of the most metabolically active
tissues in the body
Alteration of diet
Immune system
Drug metabolism Gut
Microbiome
Antibiotic use
Allergy obesity
Formation
of gall stone
8. ? What endogenous metabolic pathway is
affected by gut microflora ?
Preparation of pseudo germ free rat
Metabolomic approach with urinary metabolites
9. Oral administration of
water, twice a day for 6
days Control Pseudo-germ free p-value
AST (IU/ℓ) 95.7 ± 18.1 114.4 ± 19.7 0.17
Control ALT (IU/ℓ) 26.6 ± 5.0 33.6 ± 10.4 0.19
ALP (IU/ℓ) 126.9 ± 25.2 153.0 ± 25.1 0.15
CPK (IU/ℓ) 77.3 ± 15.2 108.3 ± 50.0 0.20
(mg/㎗) 0.1 ± 0.0 0.1 ± 0.0 0.92
Plasma, T-BIL
Urine GLU
(mg/㎗) 144.9 ± 27.6 154.7 ± 38.8 0.65
(mg/㎗) 100.5 ± 6.4 103.8 ± 8.5 0.50
CHO
Pseudogerm-free
(mg/㎗) 42.8 ± 11.9 42.5 ± 13.6 0.98
TG
After 2 day PRO (g/㎗) 5.6 ± 0.1 5.6 ± 0.5 0.88
withdraw of drug ALB (g/㎗) 3.0 ± 0.1 2.9 ± 0.2 0.64
A/G 1.1 ± 0.0 1.1 ± 0.1 0.56
Antibiotic treatment
(200 mg/kg for each (mg/㎗) 18.9 ± 4.2 19.5 ± 6.2 0.85
BUN
antibiotics, twice a day
for 6 days) (mg/㎗) 0.4 ± 0.0 0.4 ± 0.0 0.21
CRE
• neomycin sulfate
• Streptomycin
• Bacitracin
10. Retention time
Positive ionization
UPLC/TOF/MS
Retention time Negative ionization
MS2 pattern
m/z
comparison of metabolic profiling
Identification of metabolites
13. Validation of analysis
QC sample Test mix
(repeatability) (repeatability, accuracy)
RT stability, peak shape, RT stability, peak shape,
detector response detector response, mass accuracy
Nat. Protocol. 2010
14. Method Validation Workflow
Preparation of QC samples and Test mix
Test mix : commercially available standards
QC samples : pooled equal aliquots of each sample
UPLC-QTOF-MS
Test mix Test mix Test mix
Nine QC QC Ten analytical samples QC Ten analytical samples QC ……… QC
Conditioning
Run order sequence
15. Summary of validation
XIC for selected ions from test mix and QC samples;
RT, peak shape, intensity and mass accuracy
No trend with time of injection observed
Process data with MarkerLynx;
Peak finding, alignment, first nine QC runs removed
Tight QC clustering prerequisite
Examine peak list table for QCs
Calculate CV% of ion intensity
Check number of ions with CV<15%, <20%, <30%
A percentage of ~70% of features with CV<30% denotes an acceptable d
ata set
25. Oxysterol and bile acids analysis by GC-MS/MS
The receptors closely related with liver toxicity
LXR (Liver X receptor),
FXR (farnesoid X receptor)
PXR (Pregnane X receptor)
The ligand metabolite for LXR, FXR, PXR: cholesterol oxysterols and bile acids
26. Metabolic pathway & related enzyme
H Cholesterol
H H
HO
Cholesterol
Cholesterol 7α-hydroxylase Cholesterol 25-hydroxylase
Cholesterol 24-hydroxylase
OH HO
OH
LXR ligand:
H H H H H Oxysterols
H H H H H H H H H H
HO HO OH HO OH HO HO
22 (R)- hydroxy cholesterol 7keto cholesterol 7hydroxy cholesterol 24(S)- hydroxy cholesterol 25-hydroxy cholesterol
CYP7B1, oxysterol 7α-hydroxylase
3β-hydroxy-∆5-C27-steroid oxidoreductase
H
Steroid H H
CYP39A1, oxysterol 7α-hydroxylase
O OH
7hydroxy-4-cholesten-3-one
Sterol 12α-hydroxylase
∆4-3-oxosteroid 5β-reductase
∆4-3-oxosteroid 5β-reductase
3α-hydroxysteroid dehydrogenase
3α-hydroxysteroid dehydrogenase
OH
O
PXR, FXR ligand: OH
OH Bile acids O
H OH
H
H H H H
H
HO OH HO OH
H
H H Chenodeoxycholic acid
7α-hydroxysteroid dehydrogenase,
Cholic acid HO OH
H 7β-hydroxysteroid dehydrogenase
7α-dehydroxylase 5-cholestane-3,7,12-triol 6β-hydroxylase
7α-dehydroxylase
O O O
OH OH OH
OH OH
OH O O
H H H
7β-dehydroxylase
H H
H H H H H H
HO OH H H H H
HO H HO OH
H OH H HO HO OH
OH
Deoxycholic acid -Muricholic acid -Muricholic acid Lithocholic acid Ursodeoxycholic acid
27. Instrumental Conditions
Analytical Instruments Oven Temp. Profile 320ºC 2 min
6890 Series Gas Chromatography
5975 Series Mass Selective Detector
1ºC/min
GC Parameter
290ºC 2 min
Column: Ultra-I (25m x 0.2mm x 0.33μm)
Injection volume: 2.0 μL
20ºC/min
Inlet mode: Split
Split Ratio 10:1 240ºC
Inlet Temp: 280ºC
Mode: Constant Flow
Flow Rate: 0.9 mL/min, He
MS Parameter
MS Source: 230ºC
MS Quad. 150ºC
Auxiliary temperature: 300°C
Solvent delay: 5 min
Acq. Mode: SIM
28. Sample preparation method
Urine 1ml
Add 10 μl of internal standard mixture
(10 μg/ ml of 5α-cholestane and d4-cholic acid)
Enzyme hyrolysis
Add 1.5 ml of sodium acetate butter (0.2 M, pH 5.2)
Add 50 μl of β-glucuronidase/aryl sulfatase
Stand at 550C at 3 hrs
Liquid-liquid extraction
Add 5 ml of t-butylmethylether
Shaking for 10 mins
Centrifuge 2500 rpm for 5 mins
Organic layer
Evaporate
Dry at P2O5/KOH
Residue
Derivatization
Add 40 μl of MSTFA/NH4I/dithioerythritol mixture GC-MS chromatogram
Stand at 600C at 30 mins
GC-MSD
Anal Biochem Submitted
29. Urinary concentration (ng/mg of creatinine) of oxysterols and bile acids in before and
after treatment of neomycin and streptomycin treated rats.
Compounds Before treatment After treatment
Mean± SD Mean± SD
63.40±36.47
7α-OH-cholesterol 53.56±22.18
120.5±67.83 166.7±140.4
7α-OH-4-cholesten-3one
13.76±2.94 18.70±9.73
7keto- cholesterol
7386±2360 5661±1606
22(R)-OH- cholesterol
ND ND
24S-OH- cholesterol
106.7±34.92 70.62±33.82*
25-OH- cholesterol
4008±1592 2866±1271*
Cholesterol
46.27±18.30 29.84±14.35*
Deoxycholic acid (DCA)
118.1±30.75 139.6±63.87
α-Muricholic acid (α-MCA)
94.96±20.28 158.9±111.3*
Chenodeoxycholic acid (CDCA)
55.10±17.04 78.58±53.64
Cholic acid (CA)
67.37±12.00 75.01±45.29
Ursodeoxycholic acid (UDCA)
81.06±38.24 56.43±24.92
Lithocholic acid (LCA)
53.91±13.58 74.43±43.61
β-Muricholic acid (α-MCA)
47.58±10.16 77.73±51.49*
5β-cholestane 3α,7α,12α-triol
*: ≤ 0.05 between before treatment and after treatment; ND: not detected
30. H 25-hydroxylase
Cytochrome p450scc H H 24-hydroxylase
HO
Cholesterol
OH HO
OH
H
Cholesterol 7α-hydroxylase H H
H H H H H H HH
HO OH HO HO
H H H H
HO HO OH 7hydroxy cholesterol 24(S)- hydroxy cholesterol 25-hydroxy cholesterol
22 (R)- hydroxy cholesterol 7keto cholesterol
CYP39A1, oxysterol 7α-
hydroxylase
3β-hydroxy-∆5-C27-steroid CYP7B1, oxysterol 7α-
oxidoreductase hydroxylase
Steroid
H
H H
O OH
7hydroxy-4-cholesten-3-one
CYP8B1,sterol 12α-hydroxylase ∆4-3-oxosteroid 5β-reductase
∆4-3-oxosteroid 5β-reductase
3α-hydroxysteroid dehydrogenase
OH 3α-hydroxysteroid dehydrogenase OH
O O
OH
H H
H H OH
H H
HO OH HO OH
H H
Cholic acid Chenodeoxycholic acid
H H 7α-hydroxysteroid
HO OH dehydrogenase,
H 6β-hydroxylase
7α-dehydroxylase 5-cholestane-3,7,12-triol 7α-dehydroxylase
O O 7β-hydroxysteroid
O
OH OH OH dehydrogenase OH
OH
OH O O
H H
H
H H
H H H H
H H
HO
HO
H
OH HO OH H H 7β-dehydroxylase H H
H OH H HO HO OH
OH
Deoxycholic acid -Muricholic acid -Muricholic acid Lithocholic acid Ursodeoxycholic acid
31. 0.014 1.8 0.025
Cholesterol 7-hydroxylase 25-hydroxylase
0.0012 Cytochrome p450sce
0.012 1.6
22(R)-OH-cholesterol/cholesterol
7-OH-cholesterol/cholesterol
25-OH-cholesterol/cholesterol
0.0010 1.4 0.008 0.020
0.006
7keto-cholesterol/cholesterol
0.010
1.2
0.008 0.0008 0.015
1.0
0.006 0.0006 0.8
0.010
0.6
0.004 0.0004
0.4 0.005
0.002
0.0002 0.2
0.000 0.0 0.000
Normal control Pseudo germ free 0.0000 Normal control Pseudo germ free Normal control Pseudo germ free
Treated groups Normal control Pseudo germ free
Treated groups Cholesterol
5-cholestane-3,7,12-triol/7-OH-4-cholesten-3-one
Treated groups
Chenodeoxycholic acid/7-OH-4-cholesten-3-one
7-OH--4-chotesten-3-one/7-OH-cholesterol
4 2.5
7-OH-4-cholesten-3-one/25-OH-cholesterol
3-OH-5-C27-steroid oxidoreductase 1.0 4
CYP7B1, oxysterol 7-hydroxylase 4-3-oxosteroid 5-reductase
3-hydroxysteroid dehydrogease
3
2.0 0.05 0.8
3
1.5 0.6
2
2
1.0 0.4
1
1
0.5 0.2
0 0.0 0.0 0
Normal control Pseudo germ free Normal control Pseudo germ free Normal control Pseudo germ free Normal control Pseudo germ free
Treated groups Treated groups Treated groups Treated groups
500 0.5 35 100
6-hydroxylase 6-hydroxylase
-muricholic acid/chenodeoxycholic acid
7-dehydroxylase
-muricholic acid/chenodeoxycholic acid
Colic acid/5-cholestane-3,7,12-triol
0.013 30 0.011 0.032
80
Deoxycholic acid/cholic acid
400 0.4
25
300 0.3 60
20
15 40
200 0.2
10
100 0.1 20
5
0 0.0 0 0
Normal control Pseudo germ free Normal control Pseudo germ free Normal control Pseudo germ free Normal control Pseudo germ free
Treated groups Treated groups Treated groups Treated groups
3.0 50
Ursodeoxycholic acid/chenodeoxycholic acid
7-hydroxysteroid dehydrogenase 7-dehydroxylase
Lithocholic acid/chenodeoxycholic acid
2.5
40
0.024
2.0
30
1.5
20
1.0
10
0.5
0.0 0
Normal control Pseudo germ free Normal control Pseudo germ free
Treated groups Treated groups
32. 160 **
Cholic acid/chenodeoxycholic acid
140
120
100
80
60
40
20
0
Normal control Pseudo germ free
Treated groups
Figure 4.
33. CONCLUSION
1) In the non targeted metabolic profiling, 20 metabolites were significantly
related to the activities of gut microbiota.
2) They are in Aromatic amino acid, Isoflavonoid metabolism & phase II
metabolism (glucuronide conjugation).
3) In the target approach on the bile acid and oxysterol, . Increase of hydroxylase
and significant decrease of 7α-dehydroxylase were observed. The urinary
concentration ratio of primary bile acids (cholic acid and chenodeoxycholic
acid), marker for hepatotoxicity, increased in pseudo germ-free condition.
4) Those findings indicated that the gut microbiota could play a significant role in
the bile acid homeostasis and liver toxicity could be happen in the absent of
gut microbiota.
5) Therefore this study provided clear clue that the gut microbiota play important
role in normal life directly and indirectly.
34. Acknowledgement
Dr. Oh Seong Kwon
Dr. Bong Chul Chung
Dr. Soo Hyun Lee
Ji Hye Ahn
Salil Bhowmik Kumar
Supported by
KFDA
KIST
MEST