• Title/Summary/Keyword: cell homogenates

Search Result 55, Processing Time 0.022 seconds

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 (용혈독소를 생산하는 기수성 비브리오균의 생리.생태적 특성과 수산식품의 위생대책 2. 해수에서 분리된 vibrio mimicus SM-9의 생리적 특성 및 저온내성)

  • 장동석;김신명;박욱연;박미연;김영만
    • Journal of Food Hygiene and Safety
    • /
    • v.12 no.1
    • /
    • pp.9-14
    • /
    • 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.

  • PDF

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

  • Kim, Su Yeon;Nguyen, Linh;Choi, Hyoung T.
    • Korean Journal of Microbiology
    • /
    • v.52 no.3
    • /
    • pp.249-253
    • /
    • 2016
  • The inky cap, Coprinellus congregatus, produces several laccase isozymes during its life cycle: both hyphal tip laccase and sclerotial laccase are involved in the fungal development. When this fungus was transferred to an acid liquid medium (pH 4.0-4.5), a new laccase was synthesized and secreted into the culture supernatant. In order to examine its regulation by external pH, green fluorescent protein gene was ligated at the downstream of the promoters having different lengths. These expression vectors having different promoter lengths were inserted into the fungal transformation vector, pBARGEM7-1. These expression vectors were introduced to the mating type a1 and a2 monokaryons, and the transformants were selected by the phosphinothricin resistance. Transformant a1 (a1TF) and transformant a2 (a2TF) were mated with each other to generate homozygotic dikaryon transformants. All these transformants were grown in neutral liquid medium for 5 days, and then the whole cell homogenates were transferred to the acidic liquid medium (pH 4.1). After 36 h incubation at $25^{\circ}C$, cells were harvested for the analysis of GFP expression. GFP expression was detected in the transformant having full-length promoter (2.0 kb), but other transformants having shorter length promoter (shorter than 1.29 kb) failed to show the fluorescence. Therefore, the acid-responsive element in the laccase promoter should be localized between -2.0 kb ~ -1.29 kb region.

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
    • /
    • v.25 no.5
    • /
    • pp.655-663
    • /
    • 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.

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

  • Lee, Seung-Mook;An, Mi-Ra;Lee, Syng-Ill;Park, Yang-Saeng
    • The Korean Journal of Physiology
    • /
    • v.17 no.1
    • /
    • pp.55-61
    • /
    • 1983
  • Effects of SITS (4-acetamido-4'-isothiocyano-2, 2'-disulfonic stilbene) on a $Na^+$ transport, tissue oxygen consumption and Na-K-ATPase activity were studied in isolated frog skin preparations. $Na^+$ transport was estimated by measuring the short-circuit current(SC) across the skin; oxygen consumption was measured in separated epidermis as well as in intact skin; and Na-K-ATPase was assayed in $24,000{\times}g$ fraction of epidermal homogenates. The SCC across the skin Was rapidly and substantially reduced in the presence of 10 mM SITS in the medium bathing the outside(mucosal) surface of the skin. When the drug was added to the inside(serosal) bathing medium, there was about 20 min delay for inhibition of SCC and the effect was less pronounced. The above effect of SITS was independent of the presence of $Cl^-$ in the bathing medium. The oxygen consumption of the skin tissue was not affected by SITS, but the Na-K-ATPase activity of a subcellular fraction of the skin was significantly inhibited. These results suggest that SITS retards $Na^+$ transport across the frog skin primarily by interfering $Na^+$ entry across the mucosal membrance of the epithelial cell, although an effect on $Na^+$ pump can not be ruled out completely.

  • PDF

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

  • Lee, Bong-Kee;Lee, Sang-Don
    • The Korean Journal of Physiology
    • /
    • v.2 no.1
    • /
    • pp.23-30
    • /
    • 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.

  • PDF