• Title/Summary/Keyword: Metabolic stability

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Sample Preparation and Stability of Human Serum and Urine Based on HPLC-DAD for Metabonomics Studies

  • Liu, Yun;Sun, Xiaoming;Di, Duolong;Feng, Yuxiang;Jin, Fengling
    • Bulletin of the Korean Chemical Society
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    • v.33 no.7
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    • pp.2156-2162
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    • 2012
  • Many literatures focus on the biological relevance and the identification of biomarkers for disease activity assessment while less attention has been paid to the development of standard procedures for sample preparation and storage based on liquid chromatography technique. The influencing factors including protein precipitation, storage temperature, storage time, and reconstitution by ultra pure water were analyzed employing HPLC-DAD. The effects were investigated from five participants over three months by principal components analysis (PCA) and the values of percent changes (PC). The samples with protein precipitation might slow the rate of bacterial enzymatic conversion. After protein precipitation, the average PC of urine samples ($0.136{\pm}0.013$, n = 5) is relatively less than that of the serum samples ($0.173{\pm}0.026$, n = 5) for three months. Minimal effects on metabolic profiles of serum and urine (PC < 0.15) are reasonable for metabolomic studies after protein precipitation and storage at $-20^{\circ}C$ for two months.

Anti-tumor activity and mitochondrial stability of disulfiram in HL-60 cells (HL-60세포에서 disulfiram의 항암작용과 미토콘드리아 안정성에 대한 연구)

  • Shin, Hyowon;Han, Yong;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.59 no.4
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    • pp.195-199
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    • 2019
  • Disulfiram (DSF) is a member of the dithiocarbamate family that can bind copper. Recent studies have shown that DSF has anti-cancer activities, but the mechanism has not been clarified. Therefore, it is important to study the action mechanism of DSF to maximize its anticancer effects. A human leukemia cell line, HL-60, was used in this study. HL-60 cells were treated with DSF and the cellular metabolic activity was measured. DSF increased the cell death of HL-60 cells in annexin V-fluorescein isothiocyanate/propidium iodide staining analysis. In addition, DSF decreased the mitochondrial membrane potential (MMP) of the HL-60 cells. The cytotoxicity of DSF on HL-60 cells was observed at 0.4 μM. Interestingly, the reduction of MMP by DSF was recovered by N-acetyl-L-cysteine, an inhibitor of reactive oxygen species (ROS) production. This suggests that the decrease in MMP by DSF is closely related to the production of ROS in HL-60 cells, which indicates the relationship between the apoptosis of HL-60 cells by DSF and the role of the mitochondria. This study provides clinicians and researchers with valuable information regarding the anti-cancer activity of DSF in terms of the action mechanism.

Microbiome-Linked Crosstalk in the Gastrointestinal Exposome towards Host Health and Disease

  • Moon, Yuseok
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.19 no.4
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    • pp.221-228
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    • 2016
  • The gastrointestinal exposome represents the integration of all xenobiotic components and host-derived endogenous components affecting the host health, disease progression and ultimately clinical outcomes during the lifespan. The human gut microbiome as a dynamic exposome of commensalism continuously interacts with other exogenous exposome as well as host sentineling components including the immune and neuroendocrine circuit. The composition and diversity of the microbiome are established on the basis of the luminal environment (physical, chemical and biological exposome) and host surveillance at each part of the gastrointestinal lining. Whereas the chemical exposome derived from nutrients and other xenobiotics can influence the dynamics of microbiome community (the stability, diversity, or resilience), the microbiomes reciprocally alter the bioavailability and activities of the chemical exposome in the mucosa. In particular, xenobiotic metabolites by the gut microbial enzymes can be either beneficial or detrimental to the host health although xenobiotics can alter the composition and diversity of the gut microbiome. The integration of the mucosal crosstalk in the exposome determines the fate of microbiome community and host response to the etiologic factors of disease. Therefore, the network between microbiome and other mucosal exposome would provide new insights into the clinical intervention against the mucosal or systemic disorders via regulation of the gut-associated immunological, metabolic, or neuroendocrine system.

Electrochemical Activation of Nitrate Reduction to Nitrogen by Ochrobactrum sp. G3-1 Using a Noncompartmented Electrochemical Bioreactor

  • Lee, Woo-Jin;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.836-844
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    • 2009
  • A denitrification bacterium was isolated from riverbed soil and identified as Ochrobactrum sp., whose specific enzymes for denitrification metabolism were biochemically assayed or confirmed with specific coding genes. The denitrification activity of strain G3-1 was proportional to glucose/nitrate balance, which was consistent with the theoretical balance (0.5). The modified graphite felt cathode with neutral red, which functions as a solid electron mediator, enhanced the electron transfer from electrode to bacterial cell. The porous carbon anode was coated with a ceramic membrane and cellulose acetate film in order to permit the penetration of water molecules from the catholyte to the outside through anode, which functions as an air anode. A non-compartmented electrochemical bioreactor (NCEB) comprised of a solid electron mediator and an air anode was employed for cultivation of G3-1 cells. The intact G3-1 cells were immobilized in the solid electron mediator, by which denitrification activity was greatly increased at the lower glucose/nitrate balance than the theoretical balance (0.5). Metabolic stability of the intact G3-1 cells immobilized in the solid electron mediator was extended to 20 days, even at a glucose/nitrate balance of 0.1.

Effect of Increasing Amounts of Ammonium Nitrogen Induced by Consecutive Mixture of Poultry Manure and Cattle Slurry on the Microbial Community during Thermophilic Anaerobic Digestion

  • Alsouleman, Khulud
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1993-2005
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    • 2019
  • Thermophilic anaerobic digestion (TAD) is characterized by higher biogas production rates as a result of assumedly faster microbial metabolic conversion rates compared to mesophilic AD. It was hypothesized that the thermophilic microbiome with its lower diversity than the mesophilic one is more susceptible to disturbances introduced by alterations in the operating factors, as an example, the supply of nitrogen-rich feedstock such as poultry manure (PM). Laboratory scaled TAD experiments using cattle slurry and increasing amounts of PM were carried out to investigate the (in-) stability of the process performance caused by the accumulation of ammonium and ammonia with special emphasis on the microbial community structure and its dynamic variation. The results revealed that the moderate PM addition, i.e., 25% (vol/vol based on volatile substances) PM, resulted in a reorganization of the microbial community structure which was still working sufficiently. With 50% PM application, the microbial community was further stepwise re-organized and was able to compensate for the high cytotoxic ammonia contents only for a short time resulting in consequent process disturbance and final process failure. This study demonstrated the ability of the acclimated thermophilic microbial community to tolerate a certain amount of nitrogen-rich substrate.

Potential of Immobilized Whole-Cell Methylocella tundrae as a Biocatalyst for Methanol Production from Methane

  • Mardina, Primata;Li, Jinglin;Patel, Sanjay K.S.;Kim, In-Won;Lee, Jung-Kul;Selvaraj, Chandrabose
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1234-1241
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    • 2016
  • Methanol is a versatile compound that can be biologically synthesized from methane (CH4) by methanotrophs using a low energy-consuming and environment-friendly process. Methylocella tundrae is a type II methanotroph that can utilize CH4 as a carbon and energy source. Methanol is produced in the first step of the metabolic pathway of methanotrophs and is further oxidized into formaldehyde. Several parameters must be optimized to achieve high methanol production. In this study, we optimized the production conditions and process parameters for methanol production. The optimum incubation time, substrate, pH, agitation rate, temperature, phosphate buffer and sodium formate concentration, and cell concentration were determined to be 24 h, 50% CH4, pH 7, 150 rpm, 30℃, 100 mM and 50 mM, and 18 mg/ml, respectively. The optimization of these parameters significantly improved methanol production from 0.66 to 5.18 mM. The use of alginate-encapsulated cells resulted in enhanced methanol production stability and reusability of cells after five cycles of reuse under batch culture conditions.

Hazardous Effect of Dietary Trans Fats on Human Health and Regulations (인체에 미치는 트랜스지방의 위해와 규제현황)

  • Gil, Bog-Im;Rho, Jeong-Hae
    • Korean journal of food and cookery science
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    • v.23 no.6
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    • pp.1015-1024
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    • 2007
  • Trans-fatty acids (TFAs) are defined as the sum of all unsaturated fatty acids that contain one or more non-conjugated double bonds in a trans configuration. Dietary trans- fatty acids originate from commercially hydrogenated oils and from dairy and meat fats. From the perspective of the food industry, partially hydrogenated vegetable oils are attractive because of their long shelf life, stability during deep-frying, and semisolidity, which can be customized to enhance the palatability of baked goods and fried foods. Although no definite differences have been documented so far between the metabolic and health effects of industrial and ruminant TFAs, the intake of industrially produced TFA has declined, and in Europe, the majority of TFAs are of ruminant origin. Due to the scientific evidence associating TFA intake with increased risk of coronary heart disease (CDH), the Korea Food and Drug Administration (KFDA) issued a final rule that requires the amount of trans fat present in foods to be declared on the nutrition label, by December 1, 2007. In addition, many food manufacturers who use partially hydrogenated oils in their products have developed, or are considering ways, to reduce or eliminate trans-fatty acids from certain food products.

Free fatty acid-induced histone acetyltransferase activity accelerates lipid accumulation in HepG2 cells

  • Chung, Sangwon;Hwang, Jin-Taek;Park, Jae Ho;Choi, Hyo-Kyoung
    • Nutrition Research and Practice
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    • v.13 no.3
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    • pp.196-204
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    • 2019
  • BACKGROUND/OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) is a common metabolic disease triggered by epigenetic alterations, including lysine acetylation at histone or non-histone proteins, affecting the stability or transcription of lipogenic genes. Although various natural dietary compounds have anti-lipogenic effects, their effects on the acetylation status and lipid metabolism in the liver have not been thoroughly investigated. MATERIALS/METHODS: Following oleic-palmitic acid (OPA)-induced lipid accumulation in HepG2 cells, the acetylation status of histone and non-histone proteins, HAT activity, and mRNA expression of representative lipogenic genes, including $PPAR{\gamma}$, SREBP-1c, ACLY, and FASN, were evaluated. Furthermore, correlations between lipid accumulation and HAT activity for 22 representative natural food extracts (NExs) were evaluated. RESULTS: Non-histone protein acetylation increased following OPA treatment and the acetylation of histones H3K9, H4K8, and H4K16 was accelerated, accompanied by an increase in HAT activity. OPA-induced increases in the mRNA expression of lipogenic genes were down-regulated by C-646, a p300/CBP-specific inhibitor. Finally, we detected a positive correlation between HAT activity and lipid accumulation (Pearson's correlation coefficient = 0.604) using 22 NExs. CONCLUSIONS: Our results suggest that NExs have novel applications as nutraceutical agents with HAT inhibitor activity for the prevention and treatment of NAFLD.

Effect of Red Ginseng with Processed Sulfur Extracts on Serum Lipids Concentration and Metabolic Variables in Diabetic Rats (홍삼의 법제유황 처리가 당뇨쥐의 혈중지질 및 대사지표물질에 미치는 영향)

  • Han, Hyun-Jung;Kim, Hae-Ja;Chong, Myong-Soo;Cho, Hwa-Eun;Choi, Yun-Hee;Lee, Ki-Nam
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.89-98
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    • 2009
  • Objectives : The purpose of this study was to evaluate effects of processed sulfur with red ginseng on streptozotocin(STZ) induced diabetic rats for expansion of processed sulfur internal application. Methods : We prepared red ginseng with non processed sulfur extracts(GS), red ginseng with processed sulfur I extracts(GPS I) and red ginseng with processed sulfur II extracts(GPS II). In the present study, we examined about contents of crdue saponin, antioxidant activity, $\alpha$-glucosidase inhibitory activity, and effects of STZ induced diabetic rats. Results : Contents of crude saponin increased by processed sulfur, and GPS II was shown highest contents in crude saponin and sulfur compared with another groups. Electron donating ability of GPS II was shown highest activity compared with GS and GPS I, SOD-like activity showed same tendency as electron donating ability at 1 $mg/m\ell$ concentration. Inhibitory activity of $\alpha$-glucosidase was approximately same level in acarbose and GPS II. Blood glucose level of GPS II group was decreased 18.34% compared with DC(diabetes control) group and maintained stability range in glucose level. but GS and GPS I showed high level compared to GPS II. Serum triglycerides concentration also showed lowest level in GPS II. The activity of ALT, AST and ALP was shown high level in diabetic induced groups, and lowest level in GPS II. Creatinine was shown non-significantly difference in each groups and GPS II was shown lowest level in BUN. Conclusions : These results suggested that processed sulfur with red ginseng have improvement effects on diabetes and internal application of processed sulfur with red ginseng have no specific toxicity in liver and kidney.

Efficient Macrocyclization for Cyclicpeptide Using Solid-Phase Reaction

  • Kim, Joong-Hup;Hong, Il-Khee;Kim, Hyo-Jeong;Jeong, Hyeh-Jean;Choi, Moon-Jeong;Yoon, Chang-No;Jeong, Jin-Hyun
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.801-806
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    • 2002
  • Cyclicpeptides are important targets in peptide synthesis because of their interesting biological properties. Constraining highly flexible linear peptides by cyclization is one of the mostly widely used approaches to define the bioactive conformation of peptides. Cyclic peptides often have increased receptor affinity and metabolic stability over their linear counterparts. We carried out virtual screening experiment via docking in order to understand the interaction between HLE-Human Leukocyte Elastase and ligand peptide and to identify the sequence that can be a target in various ligand peptides. We made cyclic peptides as a target base on Metlle-Phe sequence having affinity for ligand and receptor active site docking. There are three ways to cyclize certain sequences of amino acids such as Met-lie-Phe-Gly-Ile. First is head-to-tail cyclization method, linking between N-terminal and C-terminal. Second method utilizes amino acid side chain such as thiol functional group in Cys, making a thioether bond. The last one includes an application of resin-substituted amino acids in solid phase reaction. Among the three methods, solid phase reaction showed the greatest yield. Macrocyclization of Fmoc-Met-Ile-Phe-Gly-Ile-OBn after cleavage of Fmoc protection in solution phase was carried out to give macrocyclic compound 5 in about 7% yield. In the contrast with solution phase reaction, solid phase reaction for macrocyclization of Met-Ile-Phe-Gly-Ile-Asp-Tentagel in normal concentrated condition gave macrocyclic compound 7 in more than 35% yield.