• 제목/요약/키워드: cell homogenates

검색결과 55건 처리시간 0.022초

용혈독소를 생산하는 기수성 비브리오균의 생리.생태적 특성과 수산식품의 위생대책 2. 해수에서 분리된 vibrio mimicus SM-9의 생리적 특성 및 저온내성 (Physiological and Ecological Characteristis of Hemolytic Vibrios and Development of Sanitary Countermeasure of Raw Fisheries Foods. 2. Physiological and Psychrotrophic Characteristics of Vibrio mimicus SM-9 Isolated from sea Water)

  • 장동석;김신명;박욱연;박미연;김영만
    • 한국식품위생안전성학회지
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    • 제12권1호
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    • pp.9-14
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    • 1997
  • Vibrio mimicus is a closely related species with V. cholerae, and has been reported to be associated with gastrointestinal infections. Although extraintestinal infections of these vibrios have also been reported in Japan and Southeast Asia. But little research papers on V. mimicus was reported in Korea. Therefore, we tried to isolate V. mimicus from the environmental sea water from April to July in Pusan, Korea. Among the isolated strains, we selected the strongest hemolytic strain and then named V. mimicus SM-9. In this paper, we checked the antibiotic susceptibility and psychrotrophic characteristics of the isolated strain. Hemolytic activity of the hemolysin produced by the isolated strain was also measured. V. mimicus was not detected from the sea water samples in April and May, but its detection rate was relatively high in June and July in Pusan, Korea. The bacteriological characteristics of V. mimicus SM-9 were Gram-negative rods, motile, oxidase positive, Voges-Proskauer negative and sucrose negative. In 23 kinds of antibiotics susceptibility test, V. mimicus SM-9 showed susceptibility to the most of antibiotics submitted while it was resistive against lincomycin, oxacillin, rifampin and vancomycin. Hemolytic activity of the hemolysin produced by V. mimicus SM-9 was highest in stationary phase of the growth curve in BHI broth at 37$^{\circ}C$ and its activity was reached 18 HU per $m\ell$ of culture supernatant. For checking the psychrotrophic property of V. mimicus SM-9, the decreasing rate of the strain in phosphate buffer solution and yellowtail flesh homogenate was examined during the storage at 4, 0, -4 and -2$0^{\circ}C$. The decreasing rates of the selected strain stored in phosphate buffer solution were greater than those in fish homogenate. Decreasing rates of V. mimicus SM-9 stored in phosphate buffer solution were not significantly different by the storage temperatures. The viable cell counts of the strain were decreased as 5 log cycles after 120 hours at all the tested temperatures. While decreasing numbers of the strain in fish homogenates were 2*4 log cycles after 120 hours. The decreasing pattern of the strain numbers were very slow after 200 hours at all the stored temperatures.

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Coprinellus congregatus의 laccase 유전자 프로모터의 산성반응인자 분석 (Analysis of an acid-responsive element in a promoter of laccase gene in the inky cap, Coprinellus congregatus)

  • 김수연;;최형태
    • 미생물학회지
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    • 제52권3호
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    • pp.249-253
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    • 2016
  • 먹물버섯의 하나인 Coprinellus congregatus는 생활사 동안 여러 종의 laccase 효소를 생성한다. 균사 끝 효소와 버섯시원체 효소 및 sclerotium (균핵) 효소들은 모두 이 균의 분화와 관련되었다. 이핵체 균사를 산성 액체배지(pH 4.0-4.5)에 접종하면 새로운 laccase가 합성되어 분비된다. 이 laccase 유전자의 프로모터의 어느 부분이 산 충격의 신호에 관련된 단백질이 결합하는가 분석하기 위하여 녹색형광단백질(green fluorescent protein, GFP) 유전자를 laccase 프로모터 2.0 kb 다음에 연결하고, 이를 형질전환 벡터인 pBARGEM7-1에 삽입함으로써 발현벡터를 구축하였다. 이 promoter-GFP 조합의 5'-region부터 차례로 제거한 짧은 길이의 이 발현벡터를 먹물버섯 교배형 a1균과 a2균에 형질전환 방법으로 도입시키고 phosphinothricin 저항성으로 형질전환체들을 선발하였다. 선발된 형질전환체 a1 (a1TF)과 a2 (a2TF)를 서로 교배하여 동형접합(homozygotic) 이핵체 형질전환체를 만들었다. 이들을 산성 액체배지에서 36시간 배양하고 균체를 모아 confocal microscope를 사용하여 형광을 분석하였다. Laccase 유전자의 전체 프로모터(2.0 kb)를 가진 발현벡터(F0-GFP)를 도입한 동형접합 형질전환체에서는 형광을 보였으나, 그 보다 짧은 길이(1.29 kb 이하)의 프로모터를 가진 형질전환체에서는 형광이 나타나지 않았다. 이 결과에 근거하여 먹물버섯의 산 충격에 대한 신호를 받는 부위가 laccase 유전자 프로모터의 -2.0 kb ~ -1.29 kb 사이에 있을 것으로 추정한다.

The Anti-Fibrogenic Effect of a Pharmaceutical Composition of[5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione] (Oltipraz) and Dimethyl-4,4′-dimethoxy-5,6,5′,6′-dimethylene dioxybiphenyl-2,2′-dicarboxylate (DDB)

  • Kang, Keon-Wook;Kim, Yoon-Gyoon;Kim, Choon-Won;Kim, Sang-Geon
    • Archives of Pharmacal Research
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    • 제25권5호
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    • pp.655-663
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    • 2002
  • Liver fibrosis is a prepathological state wherein damaged liver tissues in chronic liver diseases, such as hepatitis, are not repaired to normal tissues, but converted to fibrous tissue. 5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione (oltipraz), a cancer chemopreventive agent, is effective against a wide variety of chemical carcinogens. Recently, we reported that oltipraz inhibits liver fibrogenesis (Kang et al., 2002). In the present study, the effects of oltipraz in combination with dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylene dioxybiphenyl-2,2'-dicarboxylate (DDb) on dimethylnitrosamine (DMN)-induced liver fibrogenesis were assessed in rats. Oltipraz (30 mg/kg body weight, po, 3 times per week for 4 weeks) was found to inhibit the increases in plasma ALT, AST and bilirubin by DMN, whereas DDB (30 mg/kg body weight, po, 3 times per week for 4 weeks) attenuated the increases in the plasma ALT and bilirubin. The lowered plasma protein and albumin contents in DMN-treated rats were completely restored by oltipraz, but not by DDB. DDB decreases liver cell injury and inflammation through inhibition of nuclear factor-kB. DMN increased the accumulation of liver collagen, as indicated by the increase in the 4-hydroxyproline content in liver homogenates, which was reduced by treatment with oltipraz, but not by DDB. Given the differential effect between oltipraz and DDB, the potential enhancement of antifibrotic efficacy by the drugs was assessed in the animal model. Despite the minimal effect of DDB on DMN-induced fibrogenesis, DDB (5-25 mg/kg), administered together with oltipraz (25-5 mg/kg), showed an additive protective effect against hepatotoxicity and fibrosis induced by DMN, which was shown by the blood chemistry parameters and histopathological analysis. The adequate composition ratio of oltipraz to DDB was 5:1. These results provide information on the pharmaceutical composition, comprising of oltipraz and DDB as the active components, for the treatment and/or prevention of liver fibrosis and cirrhosis.

개구리 피부의 Sodium 이동, 산소 소모량 및 Na-K-ATPase에 대한 SITS의 영향 (Effects of SITS on Sodium Transport, Oxygen Consumption and Na-K-ATPase of the Frog Skin)

  • 이승묵;안미라;이승일;박양생
    • The Korean Journal of Physiology
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    • 제17권1호
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    • pp.55-61
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    • 1983
  • 적출된 개구리 피부에서 $Na^+$이동, 산소소모량 및 Na-K-ATPase활성도에 대한 SITS(4-acetamido-4'-isothiocyano-2, 2'-disulfonic stilbene)의 영향을 연구하였다. 피부를 통한 능동적$Na^+$이동을 추정하기 위하여 short-circuit current(SCC)를 측정하였으며, 산소소모량은 피부조직 및 분리된 표피조직에서 측정하였으며, Na-K-ATPase활성도는 표피조직의 $24,000{\times}g$분획에서 측정하였다. 피부를 통한 SCC는 10 mM SITS가 피부외측용액에 첨가될 때 급격히 하강하였으며, 내측용액에 첨가될 때는 20분정도 지난후 하강하기 시작하였으나 그 하강정도는 전자에 비해 약했다. SITS에 의한 SCC억제현상은 용액내에 $Cl^-$이 없을때도 나타났다. SITS에 의하여 피부 및 표피조직의 산소소모량은 억제되지 않았으나 표피조직분획내 Na-K-ATPase활성도는 심하게 억제되었다. 이상과 같은 성적은 SITS가 개구리 피부에서 능동적 $Na^+$이동을 강력히 억제함을 나타내는데, 이러한 억제작용은 이 약물이 주로 상피세포의 외측막에 작용하여 나타나는 것으로 사료되지만 $Na^+$펌프를 억제할 가능성을 전연 배제할 수는 없다.

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각종 인체 암조직의 당의 산화경로 분석 (Oxidative Pathway of $C^{14}-glucose$ in Various Human Cancer Tissues)

  • 이봉기;이상돈
    • The Korean Journal of Physiology
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    • 제2권1호
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    • pp.23-30
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    • 1968
  • Tissue homogenates of 12 kinds of human cancer tissues were incubated separately in medium containing $C^{14}-1-glucose$ and $C^{14}-6-glucose$ as a substrate in order to observe the oxidative pathway of glucose in the tumor tissues. At the end of 3 hours incubation in the Dubnuff metabolic shaking incubator, respiratory $CO_2$ samples trapped by alkaling which was placed in the center well of incubation flask were analysed for total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate and pyruvate. Calculations were made on the glucose consumption rate and accumulation rates of lactate and pyruvate. Fractionation of oxidative pathway of glucose was carried out by calculating $C^{14}O_2 yields from C-1 and C-6 carbon of glucose. The following results were obtained. 1. In 12 kinds of human cancer, total $CO_2$ production rates were less than $8{\mu}M/gm$ except 2 cases. These lower values impressed that oxidative metabolism in the tumor tissues generally inhibited as compared with that in normal tissues. On the other hand, fractions of $CO_2$ derived from glucose to total $CO_2$ production rates (RSA) were less than 10% in every case. These facts showed that oxidation of glucose into $CO_2$ was remarkably inhibited in the tumor tissues. 2. Factions of glucose disappeared into $CO_2\;(RGD_{CO_2})$, lactate $(RGD_L)$, pyruvate $(RGD_P)$ to glucose consumption rates were as follows. $RGD_{CO_2}$ were less than 2% in cases of in this experiment and $RGD_L$ showed more than 5% except in 2 cases. These facts showed that anaerobic degradation of glucose into 3 carbon compounds was easily proceeded but further degradation into $CO_2$ via the TCA cycle was greatly inhibited resulting in accumulation of lactate. There are large variation in values of $RGD_P$ in different kinds of tumor tissue but relatively higher values in $RGD_{CO_2}$ were obtained in the tumor tissues as compared with those of normal tissues. 3. The oxidative pathway of glucose in tumor tissues were analyzed from the values of RSA which were obtained in $C^{14}-1\;and\;C^{14}-6-glucose$ incubation experiments. It was found that 3% of $CO_2$ derived from glucose were oxidized via the principal EMP-TCA cycle and the remainder were via alternate pathway such as HMP in the liver cancer and values in other cancer tissues were as follows; 4% in the tongue cancer, 6% in the colon cancer, 6% in the lung cancer, 9% in the stomach cancer, 11% in the ovarian cancer, 12% in the neck tumor, 22% in the uterine cancer, 22% in the bladder tumor, 32% in the spindle cell sarcoma and 65% in the brain tumor. These values except later 2 cases showed less than 30% which is the lowest value among the normal tissues. Even in the brain tumor in which showed highest value in the tumor group. It is reasonable to suppose that this fraction was remarkably decreased because values in normal brain tissue was more than 90%. From the above data, it was concluded that in tumor tissues, oxidation of glucose via TCA cycle was greatly inhibited but correlation between degree of inhibited oxidation of glucose via TCA cycle and malignancy of tumor were not clarified in this experiments.

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