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.
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 summarizes research on the purification, characterization, and antifungal activity of a chitinase enzyme from Streptomyces venezuelae P10. Key findings include:
1) The chitinase was purified using ammonium sulfate precipitation, chitin affinity chromatography, and DEAE-cellulose anion exchange chromatography, yielding a purified enzyme with a molecular weight of 66 kDa.
2) The purified chitinase demonstrated optimal activity at 35°C and pH 6-8 and showed antifungal activity against phytopathogens such as Aspergillus niger and Alternaria alternate.
3) Thin layer chromatography analysis identified the chitin
This document summarizes a study examining the toxicity and antioxidant activity of two extracts from the brown seaweed Fucus vesiculosus. Both extracts were found to lack relevant toxic effects in acute and 4-week toxicity tests in rats. The extracts exhibited antioxidant activity in non-cellular systems by reducing oxidative stress and scavenging free radicals. They also showed antioxidant effects in activated macrophages and in the plasma and erythrocytes of rats treated with the extracts for 4 weeks, indicating their compounds may be absorbed and act as antioxidants in vivo. The findings support the view that daily consumption of one extract could benefit humans by reducing oxidative stress.
Application of Capillary Electrophoresis in Follow-up of Fermentation and Cel...François de l'Escaille
This document describes how capillary electrophoresis can be used to monitor various analytes during fermentation and cell culture processes. It discusses methods for analyzing carbohydrates, organic acids, cations, and proteins using a Beckman Coulter PA 800 plus instrument. Various capillary electrophoresis kits are used to separate and detect analytes like glucose, lactic acid, ammonium, and IgG, allowing monitoring of nutrients, metabolites, and therapeutic proteins over time. Results are presented showing analysis of samples from an mAb production process run in different media. Capillary electrophoresis is concluded to be a useful tool for process monitoring with minimal sample preparation.
Evaluation of Metabolic Stability of Kinsenoside, an Antidiabetic Candidate, ...Cây thuốc Việt
This study evaluated the metabolic stability of kinsenoside, a potential antidiabetic drug candidate, in rat and human liver microsomes. Kinsenoside was found to be stable in buffer and showed little non-enzymatic degradation over 8 hours. When incubated with liver microsomes, 74-78% of kinsenoside remained after 2 hours, indicating good metabolic stability. These results suggest kinsenoside would exert its pharmacological effects as the parent compound rather than as metabolites, and provide useful information for further pharmacokinetic studies.
This document discusses strategies for testing nanomaterials. It begins by introducing Maria Dusinska from the Norwegian Institute for Air Research as the author. It then discusses several key points:
- The importance of characterizing nanomaterials' physicochemical properties which influence biological interactions.
- Recommendations for adapting common toxicity assays like the cytokinesis-blocked micronucleus assay to accurately assess nanomaterials, since they can interfere with assay components.
- Results from the NanoTEST consortium evaluating various nanomaterials in different assays, finding some caused genotoxic effects depending on cell type and dispersion protocol.
- The need to represent in vivo exposure conditions like serum content to understand uptake and effects of nan
This document summarizes a study examining the absorption, bioavailability, and metabolism of oral and intravenous resveratrol in human volunteers. The key findings were:
- Absorption of a 25 mg oral dose was at least 70%, with peak plasma levels of resveratrol and metabolites of 491 ng/ml reached after 1 hour. However, only trace amounts of unchanged resveratrol (<5 ng/ml) were detected in plasma.
- Most of the oral dose was recovered in urine. Metabolism via sulfate conjugation, glucuronic acid conjugation, and hydrogenation of the aliphatic double bond was identified.
- Sulfate conjugation in the intestine/
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 summarizes research on the purification, characterization, and antifungal activity of a chitinase enzyme from Streptomyces venezuelae P10. Key findings include:
1) The chitinase was purified using ammonium sulfate precipitation, chitin affinity chromatography, and DEAE-cellulose anion exchange chromatography, yielding a purified enzyme with a molecular weight of 66 kDa.
2) The purified chitinase demonstrated optimal activity at 35°C and pH 6-8 and showed antifungal activity against phytopathogens such as Aspergillus niger and Alternaria alternate.
3) Thin layer chromatography analysis identified the chitin
This document summarizes a study examining the toxicity and antioxidant activity of two extracts from the brown seaweed Fucus vesiculosus. Both extracts were found to lack relevant toxic effects in acute and 4-week toxicity tests in rats. The extracts exhibited antioxidant activity in non-cellular systems by reducing oxidative stress and scavenging free radicals. They also showed antioxidant effects in activated macrophages and in the plasma and erythrocytes of rats treated with the extracts for 4 weeks, indicating their compounds may be absorbed and act as antioxidants in vivo. The findings support the view that daily consumption of one extract could benefit humans by reducing oxidative stress.
Application of Capillary Electrophoresis in Follow-up of Fermentation and Cel...François de l'Escaille
This document describes how capillary electrophoresis can be used to monitor various analytes during fermentation and cell culture processes. It discusses methods for analyzing carbohydrates, organic acids, cations, and proteins using a Beckman Coulter PA 800 plus instrument. Various capillary electrophoresis kits are used to separate and detect analytes like glucose, lactic acid, ammonium, and IgG, allowing monitoring of nutrients, metabolites, and therapeutic proteins over time. Results are presented showing analysis of samples from an mAb production process run in different media. Capillary electrophoresis is concluded to be a useful tool for process monitoring with minimal sample preparation.
Evaluation of Metabolic Stability of Kinsenoside, an Antidiabetic Candidate, ...Cây thuốc Việt
This study evaluated the metabolic stability of kinsenoside, a potential antidiabetic drug candidate, in rat and human liver microsomes. Kinsenoside was found to be stable in buffer and showed little non-enzymatic degradation over 8 hours. When incubated with liver microsomes, 74-78% of kinsenoside remained after 2 hours, indicating good metabolic stability. These results suggest kinsenoside would exert its pharmacological effects as the parent compound rather than as metabolites, and provide useful information for further pharmacokinetic studies.
This document discusses strategies for testing nanomaterials. It begins by introducing Maria Dusinska from the Norwegian Institute for Air Research as the author. It then discusses several key points:
- The importance of characterizing nanomaterials' physicochemical properties which influence biological interactions.
- Recommendations for adapting common toxicity assays like the cytokinesis-blocked micronucleus assay to accurately assess nanomaterials, since they can interfere with assay components.
- Results from the NanoTEST consortium evaluating various nanomaterials in different assays, finding some caused genotoxic effects depending on cell type and dispersion protocol.
- The need to represent in vivo exposure conditions like serum content to understand uptake and effects of nan
This document summarizes a study examining the absorption, bioavailability, and metabolism of oral and intravenous resveratrol in human volunteers. The key findings were:
- Absorption of a 25 mg oral dose was at least 70%, with peak plasma levels of resveratrol and metabolites of 491 ng/ml reached after 1 hour. However, only trace amounts of unchanged resveratrol (<5 ng/ml) were detected in plasma.
- Most of the oral dose was recovered in urine. Metabolism via sulfate conjugation, glucuronic acid conjugation, and hydrogenation of the aliphatic double bond was identified.
- Sulfate conjugation in the intestine/
This document discusses using an aqueous two-phase system (ATPS) and membrane separation for integrated clarification and purification of monoclonal antibodies from cell culture broth. ATPS uses the natural separation of aqueous solutions into two immiscible phases to extract monoclonal antibodies into one phase while leaving impurities in the other. A new membrane-based separator is presented that can selectively separate the antibody-containing phase through modification with surfactants. Testing of various ATPS compositions identified one that extracted over 90% of antibodies from model systems. This optimized ATPS coupled with membrane separation achieved 78% antibody recovery from actual cell culture broth while significantly reducing DNA and host cell protein impurities.
In-vitro antioxidant and GC-MS analysis ethanolic extract of poly herbal drugSkyfox Publishing Group
Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against
infections and degenerative diseases. Current research is now directed towards natural antioxidants originated from plants due to safe
therapeutics. Poly herbal drugs is used in Indian traditional medicine for a wide range of various ailments. To understand the mechanism
of pharmacological actions, antioxidant properties of the Poly herbal drugs extract were tested using standard in vitro models. The
ethanolic extract of Poly herbal drugs exhibited strong scavenging effect on superoxide, nitric oxide radical and reducing power radical
scavenging assay. The free radical scavenging effect of Poly herbal drugs extract was comparable with that of the reference antioxidants.
The data obtained in the present study suggests that the extract of Poly herbal drugs have potent Invitro antioxidant and Anti Diabetic
activity against free radicals, prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage.
Bio assay guided isolation identification active constituents of walnut leav...Debanjan Chatterjee
This Presentation revolves round about the isolation and the characterization of the active molecule megastimene which has a power full activity as anti hyperglycemic ..and they have furthur processed it for formulation
This summary provides the key points from the document in 3 sentences:
This study explored how sulforaphane affects calcium levels and cell viability in renal tubular cells. The results showed that sulforaphane increased calcium levels in the cells in a concentration-dependent manner by promoting calcium entry from outside the cell rather than calcium release from internal stores. Sulforaphane also decreased cell viability in a concentration-dependent manner, which was independent of changes in calcium levels and may have involved the induction of apoptosis.
The kidneys filter blood every 5 minutes, exposing glomerular cells to constant stimuli. The document discusses communication between glomerular cells, specifically podocytes, which is essential for proper glomerular function. Studies show podocytes resemble neurons and possess glutamatergic vesicles containing Rab3A. Blocking NMDA receptors in vitro and in vivo impairs podocytes and increases albumin permeability, suggesting glutamate signaling regulates podocyte homeostasis.
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
Sample Final Report_ Extraction of CYP microsomesEric Sudar
This document describes a proposed method to extract microsomes containing cytochrome P450 (CYP) enzymes from human liver homogenate using super-paramagnetic microspheres coated with anti-CYP2D6 antibodies. Current extraction methods require expensive centrifugation equipment not available in most forensic labs. The method aims to overcome viscosity issues when recovering the magnetic beads from liver homogenate. Bovine serum albumin is coupled to beads to determine the amount of protein coupled to the bead surface, as a proxy for quantifying antibody binding. The recovered beads will be analyzed by liquid chromatography-mass spectrometry and flow cytometry to assess anti-CYP2D6 IgG binding to CYP2D6.
This study developed a new fluorescence-based assay to quantify endosome fusion in living cells. The assay uses BODIPY-labeled avidin, which exhibits a 10-fold increase in fluorescence upon binding to biotin. BHK fibroblasts were pulse-labeled with BODIPY-avidin and a red fluorescent marker. After specified chase times, a second cohort of endosomes was pulse-labeled with biotin-conjugated probes. Fusion was detected by increased BODIPY fluorescence in individual endosomes, measured by ratio imaging. Applying this assay, the study found that over 90% of avidin-labeled endosomes fused within 10 minutes, with fusion decreasing at longer chase times, indicating endosome
This document describes a study that integrated multiple genotoxicity endpoints, including the Pig-a assay, into a standard 28-day rodent toxicity study with urethane. Male rats were treated with urethane at doses of 25, 100, and 250 mg/kg/day or a vehicle control for 29 days. Additional endpoints measured were micronucleated reticulocytes and polychromatic erythrocytes, chromosome aberrations, and DNA damage via comet assay in blood and various tissues. Ethyl methanesulfonate was used as a positive control. Statistically significant increases were observed for urethane in Pig-a mutant reticulocytes and red blood cells
A visual chip-based coimmunoprecipitation technique for analysis of protein–p...Qing Chen
This document describes a visual chip-based coimmunoprecipitation (vChip-coIP) technique for analyzing protein-protein interactions. Key points:
1. The technique combines advantages of antibody microarrays, traditional coIP, and silver enhancement detection. Antibodies are spotted onto slides to capture interacting protein pairs from cell lysates.
2. Interactions are detected using a biotinylated antibody, colloidal gold-labeled streptavidin, and silver enhancement. This makes interaction signals visible without further processing.
3. The technique is shown to be simple, cost-effective, and efficient for comprehensive study of protein-protein interactions using small amounts of crude cell lysate.
This document describes a new method called multiple products monitoring (MpM) for quantifying target peptides using liquid chromatography-mass spectrometry (LC-MS). MpM monitors multiple product ions from the MS2 spectra of target peptides, rather than selecting a single product ion as in selected/multiple reaction monitoring. It uses a scoring system based on the intensities of matching product ions between the target and reference MS2 spectra. The authors demonstrate that MpM improves sensitivity and selectivity for peptide quantification compared to conventional approaches.
Tracer techniques involve using labeled compounds, such as radioactive isotopes, to trace intermediates and steps in biosynthetic pathways in plants. Labeled precursors are introduced into plants and their movement and incorporation into products is tracked over time. This allows investigation of precursor-product relationships, localization of compounds, and elucidation of biosynthetic sequences. Tracer techniques have various applications, including studying the biosynthesis of secondary metabolites, photosynthesis, and mineral uptake in plants.
CYP2A6_HPLC_PK_2015 New Simple Method for Coumarin in Liver Cytochrome of RatsWael Ebied
This document describes the development and validation of a new liquid chromatography method for determining coumarin and its 7-hydroxy metabolite as a marker of cytochrome P450 2A6 activity in rats. The method uses a C18 column and isocratic mobile phase to separate coumarin, 7-hydroxycoumarin, and an internal standard. The method was validated and showed good linearity, precision, and accuracy. This validated method was then applied to study the effect of resveratrol, sulforaphane, and thymoquinone on hepatic CYP2A6 activity in spontaneously hypertensive rats.
This document describes a study that screened a plant extract library to identify inhibitors of signal transduction pathways mediated by the cholecystokinin receptor CCK1. Researchers developed a beta-lactamase gene reporter assay to detect plant extracts that inhibit CCK1 receptor signaling. They performed high-throughput screening of over 7,000 plant extracts and identified hits using the reporter assay. Researchers then isolated the active compounds from hits and characterized their mechanism of action, identifying a novel selective inhibitor of Gαq/11-coupled receptors.
The document summarizes the biochemical and biophysical characterization of AnAEst, a novel SGNH hydrolase. Key points:
1) AnAEst is characterized as an arylesterase that hydrolyzes small chain fatty acid aryl esters, with optimal activity at pH 7.5 and temperatures of 25-45°C.
2) Active site residues serine 17, arginine 54, and leucine 86 were selected for mutagenesis studies. Mutants showed altered substrate specificity and kinetic parameters compared to the wild-type.
3) Biophysical analysis found that AnAEst is most stable at pH 5.5 and temperatures of 25-45°C, with half
Moringa is a plantfood of high nutritional value, ecologically and economically beneficial and readily available in the countries hardest hit by the food crisis. http://miracletrees.org/ http://moringatrees.org/
Histopathological effects of nanosilver (Ag-NPs) in liver after dermal exposu...Nanomedicine Journal (NMJ)
Objective(s):
With the advent of nanotechnology, significant progress has been made in the area of nanoscale materials such as nanosilver (Ag-Nps). These nanoparticles have a wide range of applications and been used for antimicrobial purposes for more than a century. However, little
attention has been paid to the toxicity of nanosilver wound dressing. This study was designed to investigate the possible histopathological toxicity of Ag-NPs in liver of mice during wound healing.
Materials and Methods:
A group of 50 female BALB/c mice of about 8 weeks were randomly divided into two groups: Ag-NPs and control groups (n=25). After creating similar wound on the backs of all animals, the wound bed was treated in Ag-NPs group, with a volume of 50 microliters of the nanosilver solution (10ppm) ,and in control group, with the same amount of distilled water. The experiment lasted for 14 days. Histopathaological samplings of liver were conducted on days 2, 7 and 14 of the experiment.
Results:
Histopathological studies demonstrated time-dependent changes in mice liver treated with Ag-NPs compared to control group. Some changes include dilation in central venous, hyperemia, cell swelling, increase of Kupffer and inflammatory cells.
Conclusion:
This study suggests that use of nanosilver for wound healing may cause a mild toxicity, as indicated by time-dependent toxic responses in liver tissue. However, this issue will have to be considered more extensively in further studies
Journal of natural products volume 64 issue, take -- triterpene saponins from...MỐc MOn
This document summarizes research on triterpene saponins isolated from Vietnamese ginseng (Panax vietnamensis) and their ability to protect liver cells. Two new dammarane-type saponins were isolated and identified, along with nine known saponins and one known sapogenin. One of the known saponins, majonoside R2, showed strong protective effects against chemically-induced liver cell death in mice. This demonstrates that the hepatoprotective properties of Vietnamese ginseng are due to dammarane saponins containing an ocotillol-type side chain, characteristic of this plant species.
Four home remedy tips for hemorrhoids are provided: 1) Increase fiber intake slowly to soften stools; 2) Avoid straining on the toilet and limit time sitting; 3) Drink water to bulk up and soften stools; 4) Avoid prolonged sitting and take breaks to change positions. The document recommends these natural remedies to help alleviate hemorrhoid symptoms.
This document discusses metabolic activation of natural products at the Dalian Institute of Chemical Physics, Chinese Academy of Sciences. It provides examples of their research on the metabolic activation of rutaecarpine, evodiamine, pyrrolizidine alkaloids, and strychnine. For each compound, it identifies the cytochrome P450 enzymes involved in bioactivation and describes efforts to characterize reactive metabolites through trapping and protein adduct formation. The research aims to better understand metabolism-mediated toxicity of natural products.
[1] Molecular inflammation is proposed as the underlying mechanism of the aging process. [2] It is driven by the accumulation of oxidative damage and redox imbalance over time, which activates NF-kB and leads to a pro-inflammatory state. [3] Calorie restriction is able to modulate this process and blunt the age-related increase in inflammatory markers.
This document discusses using an aqueous two-phase system (ATPS) and membrane separation for integrated clarification and purification of monoclonal antibodies from cell culture broth. ATPS uses the natural separation of aqueous solutions into two immiscible phases to extract monoclonal antibodies into one phase while leaving impurities in the other. A new membrane-based separator is presented that can selectively separate the antibody-containing phase through modification with surfactants. Testing of various ATPS compositions identified one that extracted over 90% of antibodies from model systems. This optimized ATPS coupled with membrane separation achieved 78% antibody recovery from actual cell culture broth while significantly reducing DNA and host cell protein impurities.
In-vitro antioxidant and GC-MS analysis ethanolic extract of poly herbal drugSkyfox Publishing Group
Antioxidants play an important role in inhibiting and scavenging free radicals, thus providing protection to human against
infections and degenerative diseases. Current research is now directed towards natural antioxidants originated from plants due to safe
therapeutics. Poly herbal drugs is used in Indian traditional medicine for a wide range of various ailments. To understand the mechanism
of pharmacological actions, antioxidant properties of the Poly herbal drugs extract were tested using standard in vitro models. The
ethanolic extract of Poly herbal drugs exhibited strong scavenging effect on superoxide, nitric oxide radical and reducing power radical
scavenging assay. The free radical scavenging effect of Poly herbal drugs extract was comparable with that of the reference antioxidants.
The data obtained in the present study suggests that the extract of Poly herbal drugs have potent Invitro antioxidant and Anti Diabetic
activity against free radicals, prevent oxidative damage to major biomolecules and afford significant protection against oxidative damage.
Bio assay guided isolation identification active constituents of walnut leav...Debanjan Chatterjee
This Presentation revolves round about the isolation and the characterization of the active molecule megastimene which has a power full activity as anti hyperglycemic ..and they have furthur processed it for formulation
This summary provides the key points from the document in 3 sentences:
This study explored how sulforaphane affects calcium levels and cell viability in renal tubular cells. The results showed that sulforaphane increased calcium levels in the cells in a concentration-dependent manner by promoting calcium entry from outside the cell rather than calcium release from internal stores. Sulforaphane also decreased cell viability in a concentration-dependent manner, which was independent of changes in calcium levels and may have involved the induction of apoptosis.
The kidneys filter blood every 5 minutes, exposing glomerular cells to constant stimuli. The document discusses communication between glomerular cells, specifically podocytes, which is essential for proper glomerular function. Studies show podocytes resemble neurons and possess glutamatergic vesicles containing Rab3A. Blocking NMDA receptors in vitro and in vivo impairs podocytes and increases albumin permeability, suggesting glutamate signaling regulates podocyte homeostasis.
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
Sample Final Report_ Extraction of CYP microsomesEric Sudar
This document describes a proposed method to extract microsomes containing cytochrome P450 (CYP) enzymes from human liver homogenate using super-paramagnetic microspheres coated with anti-CYP2D6 antibodies. Current extraction methods require expensive centrifugation equipment not available in most forensic labs. The method aims to overcome viscosity issues when recovering the magnetic beads from liver homogenate. Bovine serum albumin is coupled to beads to determine the amount of protein coupled to the bead surface, as a proxy for quantifying antibody binding. The recovered beads will be analyzed by liquid chromatography-mass spectrometry and flow cytometry to assess anti-CYP2D6 IgG binding to CYP2D6.
This study developed a new fluorescence-based assay to quantify endosome fusion in living cells. The assay uses BODIPY-labeled avidin, which exhibits a 10-fold increase in fluorescence upon binding to biotin. BHK fibroblasts were pulse-labeled with BODIPY-avidin and a red fluorescent marker. After specified chase times, a second cohort of endosomes was pulse-labeled with biotin-conjugated probes. Fusion was detected by increased BODIPY fluorescence in individual endosomes, measured by ratio imaging. Applying this assay, the study found that over 90% of avidin-labeled endosomes fused within 10 minutes, with fusion decreasing at longer chase times, indicating endosome
This document describes a study that integrated multiple genotoxicity endpoints, including the Pig-a assay, into a standard 28-day rodent toxicity study with urethane. Male rats were treated with urethane at doses of 25, 100, and 250 mg/kg/day or a vehicle control for 29 days. Additional endpoints measured were micronucleated reticulocytes and polychromatic erythrocytes, chromosome aberrations, and DNA damage via comet assay in blood and various tissues. Ethyl methanesulfonate was used as a positive control. Statistically significant increases were observed for urethane in Pig-a mutant reticulocytes and red blood cells
A visual chip-based coimmunoprecipitation technique for analysis of protein–p...Qing Chen
This document describes a visual chip-based coimmunoprecipitation (vChip-coIP) technique for analyzing protein-protein interactions. Key points:
1. The technique combines advantages of antibody microarrays, traditional coIP, and silver enhancement detection. Antibodies are spotted onto slides to capture interacting protein pairs from cell lysates.
2. Interactions are detected using a biotinylated antibody, colloidal gold-labeled streptavidin, and silver enhancement. This makes interaction signals visible without further processing.
3. The technique is shown to be simple, cost-effective, and efficient for comprehensive study of protein-protein interactions using small amounts of crude cell lysate.
This document describes a new method called multiple products monitoring (MpM) for quantifying target peptides using liquid chromatography-mass spectrometry (LC-MS). MpM monitors multiple product ions from the MS2 spectra of target peptides, rather than selecting a single product ion as in selected/multiple reaction monitoring. It uses a scoring system based on the intensities of matching product ions between the target and reference MS2 spectra. The authors demonstrate that MpM improves sensitivity and selectivity for peptide quantification compared to conventional approaches.
Tracer techniques involve using labeled compounds, such as radioactive isotopes, to trace intermediates and steps in biosynthetic pathways in plants. Labeled precursors are introduced into plants and their movement and incorporation into products is tracked over time. This allows investigation of precursor-product relationships, localization of compounds, and elucidation of biosynthetic sequences. Tracer techniques have various applications, including studying the biosynthesis of secondary metabolites, photosynthesis, and mineral uptake in plants.
CYP2A6_HPLC_PK_2015 New Simple Method for Coumarin in Liver Cytochrome of RatsWael Ebied
This document describes the development and validation of a new liquid chromatography method for determining coumarin and its 7-hydroxy metabolite as a marker of cytochrome P450 2A6 activity in rats. The method uses a C18 column and isocratic mobile phase to separate coumarin, 7-hydroxycoumarin, and an internal standard. The method was validated and showed good linearity, precision, and accuracy. This validated method was then applied to study the effect of resveratrol, sulforaphane, and thymoquinone on hepatic CYP2A6 activity in spontaneously hypertensive rats.
This document describes a study that screened a plant extract library to identify inhibitors of signal transduction pathways mediated by the cholecystokinin receptor CCK1. Researchers developed a beta-lactamase gene reporter assay to detect plant extracts that inhibit CCK1 receptor signaling. They performed high-throughput screening of over 7,000 plant extracts and identified hits using the reporter assay. Researchers then isolated the active compounds from hits and characterized their mechanism of action, identifying a novel selective inhibitor of Gαq/11-coupled receptors.
The document summarizes the biochemical and biophysical characterization of AnAEst, a novel SGNH hydrolase. Key points:
1) AnAEst is characterized as an arylesterase that hydrolyzes small chain fatty acid aryl esters, with optimal activity at pH 7.5 and temperatures of 25-45°C.
2) Active site residues serine 17, arginine 54, and leucine 86 were selected for mutagenesis studies. Mutants showed altered substrate specificity and kinetic parameters compared to the wild-type.
3) Biophysical analysis found that AnAEst is most stable at pH 5.5 and temperatures of 25-45°C, with half
Moringa is a plantfood of high nutritional value, ecologically and economically beneficial and readily available in the countries hardest hit by the food crisis. http://miracletrees.org/ http://moringatrees.org/
Histopathological effects of nanosilver (Ag-NPs) in liver after dermal exposu...Nanomedicine Journal (NMJ)
Objective(s):
With the advent of nanotechnology, significant progress has been made in the area of nanoscale materials such as nanosilver (Ag-Nps). These nanoparticles have a wide range of applications and been used for antimicrobial purposes for more than a century. However, little
attention has been paid to the toxicity of nanosilver wound dressing. This study was designed to investigate the possible histopathological toxicity of Ag-NPs in liver of mice during wound healing.
Materials and Methods:
A group of 50 female BALB/c mice of about 8 weeks were randomly divided into two groups: Ag-NPs and control groups (n=25). After creating similar wound on the backs of all animals, the wound bed was treated in Ag-NPs group, with a volume of 50 microliters of the nanosilver solution (10ppm) ,and in control group, with the same amount of distilled water. The experiment lasted for 14 days. Histopathaological samplings of liver were conducted on days 2, 7 and 14 of the experiment.
Results:
Histopathological studies demonstrated time-dependent changes in mice liver treated with Ag-NPs compared to control group. Some changes include dilation in central venous, hyperemia, cell swelling, increase of Kupffer and inflammatory cells.
Conclusion:
This study suggests that use of nanosilver for wound healing may cause a mild toxicity, as indicated by time-dependent toxic responses in liver tissue. However, this issue will have to be considered more extensively in further studies
Journal of natural products volume 64 issue, take -- triterpene saponins from...MỐc MOn
This document summarizes research on triterpene saponins isolated from Vietnamese ginseng (Panax vietnamensis) and their ability to protect liver cells. Two new dammarane-type saponins were isolated and identified, along with nine known saponins and one known sapogenin. One of the known saponins, majonoside R2, showed strong protective effects against chemically-induced liver cell death in mice. This demonstrates that the hepatoprotective properties of Vietnamese ginseng are due to dammarane saponins containing an ocotillol-type side chain, characteristic of this plant species.
Four home remedy tips for hemorrhoids are provided: 1) Increase fiber intake slowly to soften stools; 2) Avoid straining on the toilet and limit time sitting; 3) Drink water to bulk up and soften stools; 4) Avoid prolonged sitting and take breaks to change positions. The document recommends these natural remedies to help alleviate hemorrhoid symptoms.
This document discusses metabolic activation of natural products at the Dalian Institute of Chemical Physics, Chinese Academy of Sciences. It provides examples of their research on the metabolic activation of rutaecarpine, evodiamine, pyrrolizidine alkaloids, and strychnine. For each compound, it identifies the cytochrome P450 enzymes involved in bioactivation and describes efforts to characterize reactive metabolites through trapping and protein adduct formation. The research aims to better understand metabolism-mediated toxicity of natural products.
[1] Molecular inflammation is proposed as the underlying mechanism of the aging process. [2] It is driven by the accumulation of oxidative damage and redox imbalance over time, which activates NF-kB and leads to a pro-inflammatory state. [3] Calorie restriction is able to modulate this process and blunt the age-related increase in inflammatory markers.
Four home remedy tips for hemorrhoids are provided: 1) Increase fiber intake slowly to soften stools; 2) Avoid straining during defecation and limit toilet time; 3) Drink water to keep bowels loose and soften stools; 4) Avoid prolonged sitting and take breaks to change positions. More information on hemorrhoid treatment can be found at the provided URL.
Four home remedy tips for hemorrhoids are provided: 1) Increase fiber intake slowly to soften stools; 2) Avoid straining during defecation and limit toilet time; 3) Drink water to keep bowels loose and soften stools; 4) Avoid prolonged sitting and take breaks to change positions. More information on hemorrhoid treatment can be found at the provided URL.
Four home remedy tips for hemorrhoids are provided: 1) Increase fiber intake slowly to soften stools; 2) Avoid straining on the toilet and limit time sitting; 3) Drink water to bulk up and soften stools; 4) Avoid prolonged sitting and take breaks to change positions. The document recommends these natural remedies to help alleviate hemorrhoid symptoms.
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.
Four home remedy tips for hemorrhoids are outlined. They include: increasing fiber intake to soften stools; avoiding straining during bowel movements and limiting time spent sitting on the toilet; drinking water to keep bowels loose; and avoiding prolonged sitting by taking breaks to change positions. The document provides advice on lifestyle and dietary changes that can help alleviate hemorrhoid symptoms.
NCTR is a research center within the FDA that conducts toxicology research to support risk assessment and regulatory decision making. It has unique facilities including primate and BSL-3 laboratories. NCTR research focuses on emerging issues like nanotechnology, nutrition, and critical path initiatives. Bioimaging technologies like microPET and MRI are used to study effects of substances like ketamine anesthesia on brain development in primates. Studies found ketamine increased neuronal cell death markers in the frontal cortex of developing monkeys.
1) Human intestinal bacteria can convert naturally-occurring plant compounds called phytoestrogens into estrogenic compounds. 2) Studies found lower rates of breast and prostate cancer in Japan and Finland where diets include soybeans and rye bread containing phytoestrogens. 3) These phytoestrogens structurally resemble the human hormone estradiol and can have estrogenic or anti-estrogenic effects depending on blood concentrations.
This document discusses research on intestinal microflora being conducted in Korea. It describes several research groups and projects investigating the roles of intestinal microflora in drug metabolism, food digestion, and human health. The goals are to better understand how microflora influence toxicity and to characterize differences in Korean microflora related to factors like diet. There is also a focus on developing diagnostic tools and establishing a Korean intestinal microflora database. The research aims to improve safety evaluation of drug metabolites and enable personalized medicine based on individual microfloral profiles.
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2009 약물대사기반 심포지엄-정호상(3)
1. Evaluation of Drug Toxicity
by Metabolom Profiling
Ho-Sang Jeong, Ph. D
Div. of Pharmacological Research, NIFDS
2. From Genes To Metabolites
GENOME
protein-gene
interactions
PROTEOME
protein-protein
interactions
METABOLISM
Bio-chemical
reactions
Citrate Cycle
3. Metabolomics: A Multidisciplinary Study
Urine
Serum
tissue
NMR spectra processed PCA
Raw NMR spectra
control STOCSY
Discovery of biomarkers disease or
Identification of metabolic pathway toxicity
4. Effort of NIFDS for Metabolomic Study
Year Budget (KWon) Contents
Metabolomics를 이용한내분비장애추정물질의 안전성 예측 방법 연구
2006 32,000 (metabolic study of putative endocrine disruptor)
식욕억제제의 약물동태 및 대사 연구
(Pharmacokinetic study of anorexic drug using metabolomics)
Metabolomics를 이용한 간독성 평가방법 개발 (Liver toxicity study using metabolomics)
NSAIDs 약물의 약물이상반응에 대한 안전성 예측 연구 (1)
(ADR study of NSAIDs using metabolomics)
2007 46,000 대사체 생체지표를 이용한 내분비계 장애작용 예측 연구
(metabolic study of putative endocrine disruptor)
And etc
Metabolomics를 이용한 신독성 평가방법 개발 (Kidney toxicity study using metabolomics)
NSAIDs 약물의 약물이상반응에 대한 안전성 예측 연구 (2)
(ADR study of NSAIDs using metabolomics)
2008 41,500
And etc
간독성평가를 위한 시험관내시험계에서의 전사체대사체 발현 분석 연구
(In vitro liver toxicity study using integrated omics technologies)
2009 37,000 NSAIDs 약물의 약물이상반응에 대한 안전성 예측 연구 (3)
(ADR study of NSAIDs using metabolomics)
And etc.
Total 156,500
8. Treatment and sample collection: Liver Toxicity
CCl4 1 ml/kg, p.o
Acetaminophen (AAP) 2 g/kg, p.o for 2 days
D-Galactosamine (GalN) 0.8 g/kg, i.p
Plasma clinical chemistry (blood)
Histopathology (liver)
Urinal NMR (urine)
0 1 2
Day
9. Study Design: Kidney Toxicity
Nephrotoxicity
Clinical Chemistry
HgCl2
Kidney weight Kidney Damage
Cis-platin
SD Histopathology
Gentamicin
p-Aminophenol
Metabolomics
- Urine Nephrotoxicity biomarker
Administration HgCl2 0.1 or 0.75 mg/kg, ip
Cis-platin (CP) 5 mg/kg for 2 days or 20 mg/kg for single, ip
Gentamicin (GEN) 5 or 160 mg/kg for 2 days, ip
p-Aminophenol (PAP) 10 or 200 mg/kg, ip
11. Biomaterial collection for PAP treatment
Plasma clinical chemistry (blood)
Histopathology (kidney)
Kidney wt
Urinal clinical chemistry (urine)
Urinal NMR (urine)
Treatment
0 1 2 3
Days
12. Import techniques and procedures in
metabonomics
NMR
Pattern recognition methods
Supervised techniques (PCA)
Unsupervised techniques (PLS-DA)
13. Histopathology: Liver
Control, Day 2 CCl4 1 ml/kg, Day 1 CCl4 1 ml/kg, Day 2
A B C
AAP 2 g/kg, Day 2 GalN 0.8 g/kg, Day 1 10
F
D E 8
**
Liver injury score
**
6
**
**
4 **
2
0
1
2
2
1
2
1
2
2
1
2
ay
ay
ay
ay
ay
ay
ay
ay
ay
ay
D
D
D
D
D
D
D
D
D
D
N
N
on
on
on
P
on
on
l4
l4
A
C
C
al
al
C
C
C
C
C
A
C
C
G
G
CCl4 AAP GalN
15. Multivariate statistical data analysis
SIMCA-P ver.11 (Umetrics, Umeå, Sweden)
Partial Least Squares (PLS) discriminant analysis
Very importance variables (VIP) were also utilized to
select putative markers for hepatotoxicity induced by
CCl4, AAP, and GalN.
16. Global profiling : Liver Toxicity
■ Day 0
● Day 1
♦ Day 2
CCl4 AAP GalN
A B C
17. Global profiling of hepatotoxicants
GalN Day1
Class 1
GalN Day5
Class 2
CCl4 Day 1
Class 9
CCl4 Day 11
Class 2
AAP Day 16
Class 2
Control (Day 0)
Class 19
23. C
C on
on _C
_C C
C l4
0.0
2.5
5.0
7.5
10.0
l4 _D
0
1
2
3
_D C ay
C ay C C
C C on l4_ 1
**
on l4_ 1 _ C Da
C y
*
_ C Da
C y l4 1
l4 1 _D
_D C ay
C ay C Cl4 2
2 on _
C Cl4
on _D _ A Da *
A y
**
_A ay P 2
A 2 _D
P
_D A ay
C A
A ay
C A on P_ 1
on P_ 1 _ G Da
y
*
Taurine
_ G Da al
y N 1
al 1 _D
N G ay
_D a
C
2-Oxoglutarate
G ay
C a on lN_ 1
_ G Da
**
on lN_ 1
y
**
al
_ G Da N 1
al y _D
N 1 G
_D ay
al
G ay N 2
al _D
N 2 ay
_D
ay 2
2
C
C on
on _C
_C C
C l4
0
1
2
3
4
5
l4 _D
0
1
2
3
4
_D C ay
C ay C C
C C on l4_ 1
on l4_ 1 _ C Da
**
_ C Da C y
C y l4 1
l4 1 _D
_D C ay
C ay C Cl4 2
C Cl4 2 on _
on _
*
_ A Da
A y
*
_ A Da
A y P 2
P 2 _D
_D A ay
A ay C A
C A on P_ 1
Citrate
on P_ 1 _G D
_G D ay
ay al
N 1
Succinate
al
N 1 _
_D G Da
G ay C a y
C a on lN_ 1
on lN_ 1 _G D
_G D ay
ay al
al N 1
N 1 _D
_ G ay
G Da
Determination of endogenous metabolites
al
al y N 2
N 2 _D
_D ay
ay 2
2
24. C C
on on
_C _C
C C
l4
0.0
0.1
0.2
0.3
l4
0
5
10
15
_D _D 20
C ay C
C 1 C
ay
C l C l4 1
on 4_
D
on _
**
_C ay _ C Da
C 1 C y
l4 l4 1
_D _D
C ay C ay
C 2 C C 2
C l4 on l4
_D
on _D
_A ay _A ay
A 2 A 2
P P
_D _D
A ay A ay
A C A
C 1 on P_ 1
Acetate
on P_
D _ G Da
_G ay al y
al 1 N 1
N _D
_D G
Betaine
ay
G ay C al
N 1
C al
N 1 on _
on _D _ G Da
_G al y
ay N 1
al
N 1 _D
_D G ay
al 2
G ay N
al
N 2 _D
_D ay
ay 2
2
C
on
C _C
on
_C C
l4
0
2
4
6
8
10
C _D
l4 C
_
0
1
2
3
4
5
ay
C C
C Da
C C y on l4_ 1
on l4 1 _ C Da
_C _D C y
l4 1
C ay _D
l4
_ 1 C ay
C Da C Cl4 2
C Cl y on _
on 4_ 2 _ A Da
_A D A y2
A ay P
P _D
_ 2 A a
A Da C A y
C on P_ 1
o n AP y 1
Lactate
_ G Da
_G _D al y
1
Allantoin
al ay N
N _D
_ 1 G a
G Da
C a C alN y 1
on lN y 1 on _
_G _D _ G Da
al y
al ay N 1
N _D
_ 1 a
G
Determination of endogenous metabolites
G Da
al y al y 2
N
N _d
*
_D 2
ay
**
ay
2
2
25. C
C on
on _C
_C C
C l4
0.0
0.1
0.2
0.3
l4 _D 0.4
0.00
0.05
0.10
0.15
0.20
_D C ay
C ay C C
C C on l4_ 1
on l4_ 1 _ C Da
*
_ C Da C y
**
C y l4 1
l4 1 _D
_D C ay
C ay C Cl4 2
C Cl4 2 on _
on _ _ A Da
_ A Da A y
A y P 2
P 2 _D
_D
A ay
A ay C A
C A on P_ 1
**
on P_ 1 _ G Da
_ G Da y
al y al
N 1
N 1
Phenylacetate
_D
_D G ay
Hippurate
G ay C a
C a on lN_ 1
on lN_ 1 _ G Da
*
_ G Da y
al y al
N 1
N 1 _D
_D G ay
G ay al
al N 2
N 2 _D
_D ay
ay ** 2
2
C
on
C _C
on
C
_C l4
0.0
0.2
0.4
0.6
C _D
l4
0
1
2
3
C ay
_D C 1
C ay C l4
C C 1
on _D
*
on l4_ _C ay
_ C Da C 1
y l4
C 1 _D
l4
_D C ay
C C 2
C
ay C l4
C l4 2 on _D
on _D _A ay
**
_A ay A 2
A P
P 2 _D
_D
A ay
A ay C A 1
C A 1 on P
on P_ _D
_G ay
_ G Da
y al
N 1
al
N 1 _D
_D
G a
G
Benzoate
ay al y 1
C al 1 C N
1-Methylnicotinamide
o n N_ on _
*
_ G Da
**
_ G Da
y al y 1
al
N 1 N
_D _D
G G a
Determination of endogenous metabolites
al ay
N al y 2
N
_4 2 _D
D
*
ay ay
2 2
26. Fingerprint of endogenous metabolites
Standard deviations from mean compound concentration
-3.0 -2.5 -2.0 -1.5 -1.0 -0.5 0.5 1.0 1.5 2.0 2.5 3.0
Compounds
Controls
Samples
Hepatotoxicant
27. Results and conclusion
□ Current study provides that urinary 1H NMR spectral based
metabolomics is a new approach to determine the change in
endogenous metabolites in animals exposed to hepatotoxicants of
CCl4, AAP, and GalN. NMR spectra showed pattern recognition
using PLS-DA through distinct separation of clustering in
hepatotoxicants-treated urine samples.
This pattern recognition may be used to screen hepatotoxic new
drug candidates in early preclinical studies.
We proposed 12 putative biomarkers to predict hepatotoxicity
induced by chemicals using NMR targeted analysis.
□ Classical clinical chemistry and histopathology provided
validation for this study and this database of biomarker patterns
may be useful for further study.
□ The suggested biomarkers should be explained how to involve in
mechanism of hepatic injury. The further study with other
hepatotoxicants needs to identify the endogenous biomarkers.
28. Histopathology : Kidney-HgCl2
Control HgCl2_0.1 mg/kg_D3 HgCl2_0.1 mg/kg_D6
a b c
d e
×100
HgCl2_0.75 mg/kg_D3 HgCl2_0.75 mg/kg_D6