• Title/Summary/Keyword: Metabolites Analysis

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Changes in the Myocardial Antioxidant Enzyme System by Post-Ischemic Reperfusion During Corontory Artery Bypass Operations (관상동맥우회술시 심근허혈후 재관류에 의한 활성산소 방어효소계의 변화)

  • 김응중;김기봉
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.850-860
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    • 1996
  • Oxygen free radicals and their metabolites have been implicated as possible causes of reperrusion injury In animal models. Their role in the clinical setting is still controversial. The aim of this study was to evaluate the degree of tissue damage, oxidative stress. and changes in the antioxidant enzyme system in patients undergoing cor nary artery bypass graft operations(CABG) with myocardial protection by cold blood cardioplegia. In patients undergoing CABG(n:10). the levels of lactate dehydrogenate(LDH), creatine phosphokinase MB fraction(CK-MB), and malondialdehyde(M DA) were measured In the coronary sinus effluent before aortic cross clamping and 20 minutes after reperfusion. At the same time, the myocardial tissue activities of superoxide dismutase(SOD). catalase(CAT), glutathione peroxiddse(GSHPX), glutathione reductase (GSSGRd), and glucose 6-phosphate dehydrogenate(GfPDH ) were determined in the right atrial auricle excised before aortic cross clamping and in the left atrial auricle excised 20 minutes after reperfuslon. The levels of increased significantly after reperrusion(p< U.05). There were no significant changes in CAT and CfPDH levels. Western blot analysis was performed to study the induction of antioxidant enzyme and demonstrated increased amount of Cu,Zn-SOD.

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32P-postlabeling Analysis of 7H-Dibenzo [c,g] carbazole and Dibenz [a,j] acridine DNA Adduct in Mice (7H-Dibenzo [c,g] carbazole과 Dibenz[a,j] acridine에 의한 DNA adduct의 32P-postlabeling 분석)

  • Roh, JH;Moon, YH;Warshawsk, D.;Talaska, G.
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.3 no.1
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    • pp.14-21
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    • 1993
  • N-Heterocyclic aromatics (NHA) are widely occurring environmental pollutants formed during the pyrolysis of nitrogen-containing organic chemicals. NAH are found in significant amounts in tobacco condensates, synthetic fuels, polluted river sediment, and effluents from the heating of coal. Following topical application 7H-dibenzo[c, g]carbazole (DBC) induces cancer in liver as well as skin, indicating that dermal exposure can lead to systemic effect. DBC and dibenz[a,j]acridine (DBA) are examples of NHA. The potency of many carcinogenic compounds is related, at least in part, to the efficiency of their biological activation. We undertook studies to determine which initial metabolites lead to the formation of high levels of carcinogen-DNA adducts in vivo. DBC and DBA's, DBA, trans-DBA-1,2-dihydrodiol (DBA-1,2-DHD), trans-DBA-3,4-dihydrodiol (DBA-3,4-DHD), and trans-DBA-5,6-dihydrodiol (DBA-5,6-DHD), were applied to the skin of mice. There were six adducts that were related to DBC application. These addusts were seen in the target organ, liver at high levels, but at very low levels in non-target organs, skin, lung and kidney. In skin, DBA produced two distinct adducts. The same two adducts were seen when DBA-3,4-DHD was applied. In addition the total adduct level elicited by DBA-3,4-DHD higher than that of parent compound. Two adducts were seen when DBA-5,6-DHD was applied, but these were very different from adducts seen with DBA. These results suggested that activation of DBA to DNA-binding compounds in skin includes initial formation of DBA-3,4-DHD.

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Effect of Panax Ginseng Saponin on Metabolism and Ion Transport in Human Erythrocytes (인삼이 적혈구세포의 해당과정 및 막 투과도에 미치는 영향)

  • Kang, Bok-Soon;Han, Kyung-Hee
    • The Korean Journal of Physiology
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    • v.17 no.2
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    • pp.125-133
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    • 1983
  • Red cell glycolytic intermediates, metabolites and metabolic ratios were studied. Glycolytic intermediates were measured in neutralized perchloric acid extracts of red cell suspensions after 3 hr incubation at $37^{\circ}C$ in the presence and absence of saponin. Adenosine triphosphate(ATP), adenosine diphosphate(ADP), pyruvate and lactate were measured by enzymatic procedures involving stoichiometric oxidation or reduction of a pyridine nucleotide. Glucose was determined using glucose oxidase after zinc hydroxide extraction. The redox state was calculated from the lactate dehydrogenase equilibrium. Adenosine triphosphatase activity(ATPase) was measured by determining the amount of phosphate released from ATP by washed erythrocyte membranes(ghost) during 20 min. incubation. Both total hydrolysis and the amount of hydrolysis that occured in the presence of ouabain were measured. The second measurement yields Mg-ATPase and represents nonspecific ATPase activity of the membranes. The difference between total and Mg-ATPase activity can be attributed to Na-K-ATPase. For the measurement of sodium fluxes, human erythrocytes were preincubated in $^{22}Na$ for 3 hr at $37^{\circ}C$, washed and suspended in a tracer-free medium. The amount of $^{22}Na$ transported out of cells at any time was determined by analysis of supernatant samples taken at various time after addition of the labeled cells to isotope-free medium. The cells and medium were separated and the radioactivity appearing in the medium was measured. From the total radioactivity in the suspension and the radioactivity appearing in the medium at known time, the rate constant for sodium release was computed. The results are summarized as follows: 1) ATP and ATP/ADP were found to increase at every concentration of saponin tested whereas ADP declined at every cone. of saponin. The increase in pyruvate and lactate were observed at every cone, of saponin and thus $NAD^+/NADH$ computed from pyruvate/lactate also increased. Glucose utilization was stimulated by saponin. 2) $Na^+-K^+-ATPase$ activities showed a biphasic response to saponin, first increasing in lower concentration and then decreasing in higher concentration of saponin. 3) The efflux of sodium was significantly increased by saponin in the range of 5 to 10 mg%. The stimulatory effect of saponin on the rate constants for active(ouabain-sensitive) sodium efflux was inhibited by addition of ouabain.

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Antimicrobial Effects and Antioxidative Activity of Baek-bu-ja(Aconiti koreani Rhizoma) by Extraction Solvent Ratio (추출용매 비에 따른 백부자(Aconiti koreani Rhizoma) 추출물의 항균효과 및 항산화효과)

  • Yoon, So-Jung;Kim, Jeung-Hoan;Lee, Kyoung-Hwan;Kwon, Hyo-Jung;Chun, Sung-Sook;Cho, Young-Je
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.258-262
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    • 2005
  • For the purpose of developing natural antioxidant, the antioxidative activity and antimicrobial of phenolics isolated from Baek-bu-ja (Aconiti koreani Rhizoma) were determined. Optimum extracting condition for phenolics were water extracts. HPLC analysis showed that the four major phenolic metabolites were rosemarinic, protocatechuic, caffeic and chlorogenic acids. The water extracts of Baekbuja did not have antimicrobial activity against H. pylori; however, the ethanol extracts revealed higher antimicrobial activity. Electron donation ability on DPPH of Baekbuja ethanol extract was 20% higher than other ethanol extracts. The 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid radical decolorization (ABTS) and antioxidant protection factor (PF) were determined with extracts from Aconiti koreani Rhizoma. 94% inhibition and 1.14 PF were shown on ABTS and antioxidant protection factor with 60% ethanol extracts. Also, TBARS (thiobarbituric acid reactive substances) showed $0.19\;{\mu}M$ in the control and $0.07\;{\mu}M$ in the 80% ethanol extracts. The result suggests that Baekbuja extract may be useful as potential sources of anti Helicobacter pylori, antioxidant.

Optimization of Culture Condition for the Hydrocinnamic Acid Production from Bacillus subtilis IJ-31 (Bacillus subtilis IJ-31에서 Hydrocinnamic Acid 생산을 위한 최적배양조건)

  • Joo, Gil-Jae;Kim, Young-Mog;Lee, Oh-Seuk;Kim, Joung-Woong;Kim, Won-Chan;Song, Kyung-Sik;Yoon, Sung-Joon;Kim, Jin-Ho;Rhee, In-Koo
    • Applied Biological Chemistry
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    • v.48 no.3
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    • pp.207-211
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    • 2005
  • The metabolites released from cultures of rhizosphere bacteria can inhibit plant growth. Bacillus subtilis IJ-31 inhibited plant growth by the production of hydrocinnamic acid (HCA). The production of HCA by plant-growth inhibiting rhizobacterium B. subtilis IJ-31 was optimized. $90.5\;{\mu}g/ml$ of HCA was obtained under the condition of 1% rice bran as carbon source, 0.5% tryptone as nitrogen source, 0.1% $ZnCl_2$ as metal source at $37^{\circ}C$ for 60 h (pH 7.0). The optimal condition for the HCA production by B. subtilis IJ-31 in the jar fermenter was established using response surface methodology (RSM) of statistical analysis system(SAS) program. The production of HCA by B. subtilis IJ-31 in the jar fermenter culture reached $102.99\;{\mu}g/ml$ when 2.24% soil extracts was added and agitation speed was 290 rpm under the same condition. And the experimental value of HCA production is $102.5\;{\mu}g/ml$ in the same culture condition. The production of HCA by B. subtilis IJ-31 is higher as 12% than that from the flask culture.

Antifouling Activity towards Mussel by Small-Molecule Compounds from a Strain of Vibrio alginolyticus Bacterium Associated with Sea Anemone Haliplanella sp.

  • Wang, Xiang;Huang, Yanqiu;Sheng, Yanqing;Su, Pei;Qiu, Yan;Ke, Caihuan;Feng, Danqing
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.460-470
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    • 2017
  • Mussels are major fouling organisms causing serious technical and economic problems. In this study, antifouling activity towards mussel was found in three compounds isolated from a marine bacterium associated with the sea anemone Haliplanella sp. This bacterial strain, called PE2, was identified as Vibrio alginolyticus using morphology, biochemical tests, and phylogenetic analysis based on sequences of 16S rRNA and four housekeeping genes (rpoD, gyrB, rctB, and toxR). Three small-molecule compounds (indole, 3-formylindole, and cyclo (Pro-Leu)) were purified from the ethyl acetate extract of V. alginolyticus PE2 using column chromatography techniques. They all significantly inhibited byssal thread production of the green mussel Perna viridis, with $EC_{50}$ values of $24.45{\mu}g/ml$ for indole, $50.07{\mu}g/ml$ for 3-formylindole, and $49.24{\mu}g/ml$ for cyclo (Pro-Leu). Previous research on the antifouling activity of metabolites from marine bacteria towards mussels is scarce. Indole, 3-formylindole and cyclo (Pro-Leu) also exhibited antifouling activity against settlement of the barnacle Balanus albicostatus ($EC_{50}$ values of 8.84, 0.43, and $11.35{\mu}g/ml$, respectively) and the marine bacterium Pseudomonas sp. ($EC_{50}$ values of 42.68, 69.68, and $39.05{\mu}g/ml$, respectively). These results suggested that the three compounds are potentially useful for environmentally friendly mussel control and/or the development of new antifouling additives that are effective against several biofoulers.

Elucidation of the Biosynthetic Pathway of Vitamin B Groups and Potential Secondary Metabolite Gene Clusters Via Genome Analysis of a Marine Bacterium Pseudoruegeria sp. M32A2M

  • Cho, Sang-Hyeok;Lee, Eunju;Ko, So-Ra;Jin, Sangrak;Song, Yoseb;Ahn, Chi-Yong;Oh, Hee-Mock;Cho, Byung-Kwan;Cho, Suhyung
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.505-514
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    • 2020
  • The symbiotic nature of the relationship between algae and marine bacteria is well-studied among the complex microbial interactions. The mutual profit between algae and bacteria occurs via nutrient and vitamin exchange. It is necessary to analyze the genome sequence of a bacterium to predict its symbiotic relationships. In this study, the genome of a marine bacterium, Pseudoruegeria sp. M32A2M, isolated from the south-eastern isles (GeoJe-Do) of South Korea, was sequenced and analyzed. A draft genome (91 scaffolds) of 5.5 Mb with a DNA G+C content of 62.4% was obtained. In total, 5,101 features were identified from gene annotation, and 4,927 genes were assigned to functional proteins. We also identified transcription core proteins, RNA polymerase subunits, and sigma factors. In addition, full flagella-related gene clusters involving the flagellar body, motor, regulator, and other accessory compartments were detected even though the genus Pseudoruegeria is known to comprise non-motile bacteria. Examination of annotated KEGG pathways revealed that Pseudoruegeria sp. M32A2M has the metabolic pathways for all seven vitamin Bs, including thiamin (vitamin B1), biotin (vitamin B7), and cobalamin (vitamin B12), which are necessary for symbiosis with vitamin B auxotroph algae. We also identified gene clusters for seven secondary metabolites including ectoine, homoserine lactone, beta-lactone, terpene, lasso peptide, bacteriocin, and non-ribosomal proteins.

Simultaneous Determination of Urinary Phytoestrogens and Estrogens by Gas Chromatography/Mass spectrometry (GC/MS에 의한 뇨 중 Phytoestrogen과 Estrogen의 동시 분석)

  • Yang, Yoon Jung;Lee, Seon Hwa;Chung, Bong Chul
    • Analytical Science and Technology
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    • v.11 no.5
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    • pp.374-385
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    • 1998
  • Phytoestrogens are biologically active compounds derived from plants foods. It had been suggested that phytoestrogens, by inhibiting aromatase in peripheral and/or cancer cells and lowering estrogen levels, may play a protective role as antipromotional compounds during growth of estrogen-dependent cancers. Therefore, simultaneous analysis of estrogens and phytoestrogens is necessary to elucidate the possible involvement of phytoestrogens in estrogen metabolism. In this view, we developed a simple and reproducible procedure to quantitatively determine estrogen and phytoestrogen metabolites. The proposed method consisted of solid phase extraction using preconditioned Serdolit AD-2 resin, enzyme hydrolysis with ${\beta}$-glucuronidase/arylsulfatase from Helix pomatia, liquid-liquid extraction and TMS-ether derivatization. And the final determination was carried out by gas chromatography/mass spectrometry (GC/MS) in selected ion monitoring mode (SIM). The precision and accuracy of this method was evaluated through within-a-day and day-to-day test. Recovery range and detection limit were 71.96~105.66%, 2~4 ng/mL, respectively. Using this method, 17 estrogen and 5 phytoestrogen compositions in urine of normal subjects were analyzed. It was found that amounts and relative distribution of urinary phytoestrigens and estrogens showed different pattern in male and female subjects.

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Characterization of Multifunctional Bacillus sp. GH1-13 (복합기능성 Bacillus sp. GH1-13 균주의 특징)

  • Kim, Sang Yoon;Sang, Mee Kyung;Weon, Hang-Yeon;Jeon, Young-Ah;Ryoo, Jae Hwan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.20 no.3
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    • pp.189-196
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    • 2016
  • Several microorganisms in particular Bacillus subtilis group have been isolated from diverse places such as soils and the gastrointestinal tract of ruminants etc., and used as biocontrol agent against various plant pathogens and utilized as plant growth promoting agents. Among them, Bacillus is well known as one of the most useful bacteria for biocontrol and plant growth promotion. Bacterium GH1-13 was isolated from a reclaimed paddy field in Wando Island and identified as Bacillus velezensis using phylogenetic analysis on the basis of 16S rRNA and gyrB gene. It was confirmed that GH1-13 produced indole acetic acid (IAA) associated with promoted growth of rice root. GH1-13 showed characteristics of antagonization against the main pathogen of rice as well as diverse pathogenic fungi. GH1-13 had biosynthetic genes, bacillomycin, bacilycin, fengycin, iturin, and surfactin which are considered to be associated closely with inhibition of growth of pathogenic fungi and bacteria. This study showed that GH1-13 could be used as a multifunctional agent for biocontrol and growth promotion of crop.

Comparison for immunophysiological responses of Jeju and Thoroughbred horses after exercise

  • Khummuang, Saichit;Lee, Hyo Gun;Joo, Sang Seok;Park, Jeong-Woong;Choi, Jae-Young;Oh, Jin Hyeog;Kim, Kyoung Hwan;Youn, Hyun-Hee;Kim, Myunghoo;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.424-435
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    • 2020
  • Objective: The study was conducted to investigate variations in the immunophysiological responses to exercise-induced stress in Jeju and Thoroughbred horses. Methods: Blood samples were collected from the jugular veins of adult Jeju (n = 5) and Thoroughbred (n = 5) horses before and after 30 min of exercise. The hematological, biochemical, and immunological profiles of the blood samples were analyzed. Blood smears were stained and observed under a microscope. The concentration of cell-free (cf) DNA in the plasma was determined using real time polymerase chain reaction (PCR). Peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells were separated using Polymorphprep, and the expression of various stress-related and chemokine receptor genes was measured using reverse transcriptase (RT) and real-time PCR. Results: After exercise, Jeju and Thoroughbred horses displayed stress responses with significantly increased rectal temperatures, cortisol levels, and muscle catabolism-associated metabolites. Red blood cell indices were significantly higher in Thoroughbred horses than in Jeju horses after exercise. In addition, exercise-induced stress triggered the formation of neutrophil extracellular traps (NETs) and reduced platelet counts in Jeju horses but not in Thoroughbred horses. Heat shock protein 72 and heat shock protein family A (Hsp70) member 6 expression is rapidly modulated in response to exercise-induced stress in the PBMCs of Jeju horses. The expression of CXC chemokine receptor 4 in PBMCs was higher in Thoroughbred horses than in Jeju horses after exercise. Conclusion: In summary, the different immunophysiological responses of Jeju and Thoroughbred horses explain the differences in the physiological and anatomical properties of the two breeds. The physiology of Thoroughbred horses makes them suitable for racing as they are less sensitive to exercise-induced stress compared to that of Jeju horses. This study provides a basis for investigating the link between exercise-induced stresses and the physiological alteration of horses. Hence, our findings show that some of assessed parameters could be used to determine the endurance performance of horses.