• Title/Summary/Keyword: strain differentiation

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Physiological Characteristics and Anti-Obesity Effect of Enterococcus faecalis MD366 isolated from Raw Milk (원유에서 분리한 Enterococcus faecalis MD366의 생리적 특성 및 비만 억제 효과)

  • Park, Sun-Young;Cho, Seong-A;Han, Noo-Ri;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.147-156
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    • 2014
  • The aim of this study was to investigate the physiological characteristics and anti-obesity effects of E. faecalis MD366 isolated from raw milk. E. faecalis MD366 inhibited lipase activity ($65.0{\pm}0.9%$) and differentiation of 3T3-L1 adipocytes ($27.4{\pm}1.4%$) at a concentration of $100{\mu}g/mL$ ($10^8CFU/g$ E. faecalis MD366). The optimum growth temperature of E. faecalis MD366 was $37^{\circ}C$. Among 16 tested antibiotics, E. faecalis MD366 demonstrated the highest sensitivity to novobiocin and the highest resistance to neomycin, kanamycin, and vancomycin. The strain also showed high acid phosphatase activity. Moreover, E. faecalis was relatively tolerant to bile juice and acid, and displayed high resistance to Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus (80.4%, 60.2%, and 65.4%, respectively). These results demonstrate that E. faecalis MD366 can be potentially used as a probiotic with anti-obesity effects.

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Antiretroviral Effects of 2',3'-Dideoxycytidine and Recombinant $Interferon-{\alpha}-A$ on the Infection of Anemia-inducing Murine Friend Virus (Anemia-inducing Murine Friend Virus 감염에 대한 2',3'-dideoxycytidine 및 $Interferon-{\alpha}-A$의 항retrovirus효과)

  • Ann, Hyung-Soo;Ahn, Ryoung-Me;Kim, Dong-Seop
    • The Korean Journal of Pharmacology
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    • v.31 no.3
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    • pp.365-375
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    • 1995
  • The anemia-inducing strain of Friend virus (FVA) is a murine retrovirus which stimulates the proliferation of erythroid progenitor cells. The progenitor cells synthesized by FVA-stimulation are unable to proceed with differentiation and accumulate in the spleen resulting in splenomegaly in infected mice. Using FVA-inoculated mice as a model, we have investigated the antiretroviral effects of 2',3'-dideoxycytidine (ddC) and recombinant $interferon-{\alpha}-A\;(rIFN-{\alpha}-A)$ on FVA infection. The extent of the infection was determined by measuring the weights of the spleens. Daily intraperitoneal injection of ddC (100 mg/kg body weight), $rIFN-{\alpha}-A$ (10 KU/mose) and the combination of both drugs to FVA inoculated mice for 18 days resulted in suppression of the growth of spleens by 15.1%, 52.7% and 61.6%, respectively. When ddC was dissolved in drinking water (0.1 mg/ml) and administered to a group of FVA inoculated mice ad libitum, and $rIFN-{\alpha}-A$ (10 KU/mouse) was intraperitoneally injected daily to another group of ddC (0.1 mg/ml) drinking mice for 18days, the growth of spleens was suppressed by 38.4% and 83.2%, respectively. These results indicate that administration of ddC via drinking water is more effective in suppressing FVA infection than the daily injection of ddC, and that the combined effects ddC and $rIFN-{\alpha}-A$ are not synergistic but additive. In order to determine whether ddC treatment alters the characteristic of the progenitor cells with respect to $Ca^{++}$ uptake, $Ca^{++}$ uptake in erythroid cells and the effect of cyclohexyladenosine (CHA) on the $Ca^{++}$ uptake were studied. $Ca^{++}$ uptake in the erythroid progenitor cells was about 20-fold greater than in mouse erythrocytes and the inhibition of $Ca^{++}$ uptake by CHA was the greatest in the progenitor cells from FVA infected mice which were treated with ddC. The inhibition was obviated by theophylline. Results of CHA binding studies showed that the erythroid progenitor cells contain both high and low affinity CHA binding sites, whereas mose erythrocytes contain only the low affinity CHA binding sites.

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Detection and Differentiation of Herpes Simplex Virus 1 and 2, and Varicella-Zoster Virus in Vesicle Fluid, Joint Fluid and Serum using PCR Method (중합효소 연쇄반응에 의한 수포액, 혈액과 관절액에서 단순포진 바이러스 1, 2와 대상포진 바이러스의 검출과 감별)

  • Park, Hae-Kyung;Woo, So-Youn;Kim, Hyun-Jin;Lee, Chung-Hwa
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.191-201
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    • 2000
  • The viruses of Herpes Simplex Virus 1 (HSV-1), Herpes Simplex Virus 2 (HSV-2) and Varicella-Zoster virus (VZV) which belong to the alpha herpes subfamily are important human pathogens. When eruptions were not fully developed from these viral infections, clinical diagnosis was not always easy and required virological confirmation test. The above viruses were reactivated in individuals who were compromised in immune competence for one reason or another. Polymerase chain reaction (PCR) enables rapid and sensitive detection of HSV and VZV DNAs. Its sensitivity was largely influenced by choice of primers. Authors conducted a study to detect of those three viruses in human specimens including vesicle fluid and joint fluid and serum using PCR methods. Primers used for this study were the general primer pair GPHV-RU which was known to amplify within the genes enjoying the highest degree of homology between UL15 of HSV and UL42 of VZV. PCR with primers hybridized pair GPHV-RU amplifies a 396 bp with THP-1 and HSV-2 standard strain DNA and 405 bp with VZV standard strain DNA. Restriction enzyme cleavage with HpaII and DdeI were used to detect and distinguish DNAs of THP-1 and HSV-2 and VZV. The purpose of this study was a rapid and easy detection of VZV and THP-1 or HSV-2 from various clinical specimens (vesicle fluid, serum and joint fluid) by PCR method. Used methods were: HSV PCR with primer 1, 2 and HpaII RE digestion; VZV nested PCR; HSV PCR with primer A, Band BssHII RE digestion. 1) In 33 cases (33/42, 78.6%) VZV was detected single or mixed infection from 42 clinical specimens which included vesicle fluid (5), serum form respiratory infected children (10), serum from immune suppressed adult cancer patients (7) and joint fluid from arthritis patients (20). 2) In 20 cases (20/42, 47.6%) HSV was detected singly or mixed infection and 19 of those cases were HSV-2 and 1 case was THP-1. 3) In 19 cases (19/42, 45.2%) VZV was singly detected which included serum from respiratory infected children (6 cases), joint fluid from arthritis patients (9 cases), vesicle fluid (2 cases) and serum form immunosuppressed cancer patients (2 cases). 4) HSV was singly detected in 6 cases (6/42, 14.3%) which included joint fluid from arthritis patients (5 cases) and serum form respiratory infected children (1 cases). 5) 14 cases of VZV and HSV mixed infection (14/42, 33.3%) were detected. They included vesicle fluid (3 cases), serum form immunosuppressed cancer patients (4 cases), serum from respiratory infected children (2 cases) and joint fluid from arthritis patients (5 cases). 6) HSV-1 and HSV-2 detection and typing by HSV PCR with primer A, Band BssHII RE digestion method was more sensitive and the results were easier to detect than on other method.

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Phylogenetic Analysis of the Genus Phellinus by Comparing the Sequences of Internal Transcribed Spacers and 5.8S Ribosomal DNA (Ribosomal DNA의 Internal Transcribed Spacer(ITS) 부위의 염기서열분석에 의한 Phellinus속의 계통분석에 관한 연구)

  • Chung, Ji-Won;Kim, Gi-Young;Ha, Myung-Gui;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.124-131
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    • 1999
  • This study was carried out to identify the phylogenetic relationship among Phellinus species by comparing the DNA sequences of the 5.8S ribosomal DNA (rDNA) and the internal transcribed spacers (ITSs), ITS1 and ITS2 regions. Two primers from the 3' end of 18S rDNA and the 5' end of 28S rDNA sequences were chosen to amplify the specific ITS regions of Phellinus spp. Phellinus strains used in the study were divided into four clusters by the phylogenetic tree based on the amplified regions of ITS and 5.8S rDNA sequences. The first cluster consist of Phellinus hartigii IMSNU 32041 and Phellinus robustus IMSNU 32068, and the second cluster consists of Phellinus linteus strains and Phellinus weirianus IMSNU 32021. Phellinus laevigatus KCTC 6229, KCTC 6230 and Phellinus igniarius KCTC 6227, KCTC 6228 belong to the third cluster. Finally, Phellinus chrysoloma KCTC 6225 and Phellinus chrysoloma KCTC 6226 are the fourth cluster. In the second cluster the differentiation between Phellinus linteus strains and Phellinus weirianus species were not possible by the comparison of the ITS sequences. These results revealed that Phellinus linteus and Phellinus weirianus cannot be established the concept of species level only by the ITS sequences. Therefore, both physiological and molecular biological methods as well as the sequences of type strains are necessary to classify the strains of these two species accurately. The comparison of the ITS sequences of four Phellinus species indicated that the sequences of the ITS1 generally are more divergent than those of the ITS2. Although the ITS sequences are varied in some species, the conserved regions in both ITS1 and ITS2 are useful tool to differentiate the species. Phellinus linteus and related species have their specific sequences in the ITS1 compared to the other species.

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Distribution and Bacteriological Characteristics of Vibrio vulnificus (Vibrio vulnificus 균의 분포 및 세균학적 특성)

  • CHANG Dong-Suck;SHIN Il-Shik;CHOI Seung-Tae;KIM Young-Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.2
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    • pp.118-126
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    • 1986
  • Vibrio vulnificus is a recently recognized halophilic organism that nay cause serious human infections. Patients infected with V. vulnificus often have a history of exposure to the sea, suggesting that the organism may be common inhabitant of marine environment. The purpose of this experiment is to investigate the distribution and bacteriological characteristics of V. vulnificus. The strain used in this experiment was isolated from sea water and sea products such as common octopus (Octopus variabilis), ark shell (Anadara broughtonii), blue crab (Ericheir japonica), and sea squirt (Synthia roretzi) collected in Pusan area from July to October in 1985. V. vulnificus was frequently isolated in August when temperature of sea water was around $26^{\circ}C$ and rarely isolated in October when temperature of sea water was around $18.5^{\circ}C$. The distinctive biochemical characteristics of V. vulnificus were ONPG hydrolysis positive and fermented lactose and not grown in peptone water contained $8\%$ NaCl. The optical density at 660 nm of the growth of V. vulnificus was reached maximum level after 8 hours of culture at $35^{\circ}C$ in brain heart infusion broth but that of V. vulnificus was little increased at $15^{\circ}C$ for 14 hours. Optimum temperature and pH for the growth of V. vulnificus were around $35^{\circ}C$ and 8.0. The specific growth rate and the generation time of V. vulnificus isolated from the samples were $1.21\;hr^{-1}$, 34 min at $35^{\circ}C$ and $0.61\;hr^{-1}$, 69 min at $25^{\circ}C$, respectively. V. vulnificus did not grow on eosin-methylene-blue agar, salmonella-shigella agar, deoxycholate agar but grew well on Endo agar, xylose-lysine-deoxycholate agar and hektoen enteric agar. On Endo agar, the colonies of V. vulnificus were red and achieved a diameter of 2 to 4 mm as a feature enabling differentiation of V. vulnificus from other Vibrio spp. V. vulnificus grow well on TCBS agar forming green colonies. V. vulnificus refrigerated at $4^{\circ}C$ exhibited a linear decline of its viablity as 1 log cycle in every 16 hours storage, while V. vulnificus freezed at $-18^{\circ}C$ almost became extinct.

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Lymphocyte Subpopulations and Proliferation of T cells, Phagocytic Activity of Leukocytes on Alcoholics (알코올중독자의 백혈구탐식능, 림프구아형 및 증식능)

  • 김용호;서병배;이정녀;김영훈
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.167-174
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    • 1996
  • Alcoholics increased susceptibility to microbial infection that is associated with decreased immunity. but there has been little experimental evidence to support alcoholics-induced increase of microbial infection directly in non-specific immunity. Therefore, we were used the method of phagocytic-plaque including all the stimulating factors for the phagocytosis, subtypes of lymphocytes and T-lymphocyte proliferation. The experimental groups were divided into 3 groups: (1) alcoholics who were hospitalized less than 1 week (newly hospitalized alcoholics), (2) alcoholics who were hospitalized more than 2 weeks (old hospitalized alcoholics), (3) healthy blood donors. We have studied 98 alcoholics and 35 healthy blood donors and control groups. A physician has checked the biological markers and diagnosed the body-condition alcoholics. The immunity and non-specific immunity on the alcoholics were analyzed by using the simultest kit and flow cytometry. Proliferation of the lymphocytes was analyzed by the phytohemmagglutinine mitogen. Phagocytosis and migration properties of leukocytes were identified on the layer formed by Staphylococcus aureus Cowan I strain. Biological markers of alcoholics and control groups, by such as blood glucose, ${\gamma}$-glutamyl transpeptidase and mean corpuscular volumes of red blood cells, were determined by biochemical and hematological methods. Compared with control groups, cluster of differentiation (CD)3+, CD8+ and CD19+ in alcoholic were more decreased except CD4+/CD8+ ratio. Proliferation of the T-lymphocytes, phagocytosis and migration properties of the leukocytes in alcoholics were decreased compared with those of control groups. According to the results observed in our experiment, they can be summerized as follows: 1, Cellular, humoral and non-specific immunities, are markedly decreased in alcoholics than those in control groups. 2. It is inferred that Phagocytic plaque formation is a very useful method to evaluate phagocytosis and migration properties of the alcoholic leukocytes 3. It is thought that the subtypes of lymphocytes, especially CD4+/CD8+ ratio, are essential methods to analyzed the alcoholic immunity. 4. Specific and non-specific immunity on the old hospitalized alcoholics was slightly increased, which depends upon the alcoholic medication.

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Cloning and Transcription Analysis of Sporulation Gene (spo5) in Schizosaccharomyces pombe (Schizosaccharomyces bombe 포자형성 유전자(spo5)의 Cloning 및 전사조절)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.112-118
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    • 2002
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.

The Establishment of Nitrogen Fixation by Cultured Cell-Rhizobium Association Through Tissue Culture Technique in Soybean (대두조직배양세포(大豆組織培養細胞) - Rhizobium에 의(依)한 질소고정력(窒素固定力))

  • Kang, Sang Jae;Park, Woo Churl
    • Current Research on Agriculture and Life Sciences
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    • v.4
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    • pp.27-35
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    • 1986
  • This experiment was carried out to elucidate the factor of nitrogenase formation and to establish the nitrogen fixation system in mixed culture of cultured cells and rhizobia through tissue culture technique using three soybean varieties, Hwangkeum, Namcheon and D 68-0099 as host plants. The results obtained were as follows; The callus was induced in embryo and radicle, but not in hypocotyl. The most favorable callus induction was caused by the individual application of 2,4-D and NAA at the concentration of 2mg/1 and 4mg/1, respectively, but in case of treating both 2,4-D and kinetin, that was done at the concentration of 0.2mg(2,4-D)/0.05mg(kinetin)per liter. The growth of cultured cell was good at the concentration of 2.0mg(2,4-D)/1 and 0.2mg(2,4-D)/0.05mg(kinetin)per liter. When cultured cells were inoculated with R. japonicum 019 and 011, their growthes were considerably inhibited. The addition of single amino acid inhibited the growth of cultured cells. Hwangkeum was inhibited considerably by methionine and leucine. The inhibition of growth by single amino acid can be abolished by the addition of certain amino acids. The differentiation of adventitious root was good at the concentration of 2.0mg 2,4-D and 0.2mg 2,4-D/0.05mg kinetin per liter. Of three host plants tested with 25 R. japonicum strains, Hwangkeum had affinity for 10 strains, Namcheon for 7 strains and D68-0099 for none. The nitrogen fixing abilities of Hwangkeum and Namcheon caused by cultured cell-Rhizobium association were high in strain 019, 007, and in 007 mixed with 119, respectively.

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