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Isolation and Identification of Histamine Degrading Bacteria from Kwamegi (과메기에서 histamine 분해능을 나타내는 세균의 분리 동정)

  • Kim Min-Woo;Kim Young-Man
    • Journal of Life Science
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    • v.16 no.1
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    • pp.120-125
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    • 2006
  • To isolate and identify histamine degrading bacteria from Kwamegi, bacteria were screened with restriction media containing histamine. Ten strains were selected through morphological and biochemical identification procedure followed by comparison with DNA sequence of 16 rRNA gene. And also, these strains were confirmed by the histamine degrading assay such as turbidity and enzymatic assay. The results of identification are as followings : Ewingella americana B791, Arthrobacter sp. R45S, Halomonas marisflava, Psychrobacter sp. 9B-7, Bacillus sp. LMC 21002, Psychrohacter cibarius BC-220, Bacillus megaterium KL-197 were identified showing homology of $99\%,\;95\%,\;98\%,\;99\%,\;99\%,\;99\%\;and\;98\%$, respectively. Three strains remain unidentified. Arthrobacter sp. R45S, H. marisflava, Bacillus sp. LMG 21002, B. megaterium KL-197 showed histamine degrading activity, whereas, Psychrobacter sp. 9B-7 only showed weak activity. Three unidentified strains also have histamine degrading activity. In contrast, E. american B791 and p. cibarius JG-220 did not show any significant activity of histamine degradation. The strains isolated from this study showed relatively fast growth rate and histamine degrading rate as compared to those from salted mackerel.

Inductive Effects of Ginseng Saponins on the Rat LDH A-gene and the Synthetic rate of Hepatocyte DNA in Regenerating Rat Liver Cells

  • Yoo, Kye-Jin;Lee, Kwang-Youl;Lee, Seung-Ki
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.58-64
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    • 1990
  • The effects of ginseng saponins, G-Rbl and G-Rc on the rat liver LDH A-gene transcnptional activity was investigated during pro-replicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl of G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 ma/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5·hours after partial hepatectomy. Dose dependent effect of G-Rbl and G-Rc (1-25 mg/100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal in- creases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc However, when the administration doses of G-Kbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rb 1 and G·Rc have stimulatory effect at the lower concentration (1 mg/100g B.W) and inhibitory effect at the higher concentration (20 moi loos 5.W) on the LDH A-gene transcription during regeneration of rat liver, Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA synthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G·Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 ug and 100 ug/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen enhancer activity for the hepatocyte proliferation during rat liver regeneration period. Keywords Inductive effects of ginsenosides, G-Rb, -Rc, and -Re, rat LDH A-gene transcription, the sin thetic rate of proteins and DNA in regeneration rat liver.

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Detection of a Microsporidium, Nosema ceranae, from Field Population of the Bumblebee, Bombus terrestris, via Quantitative Real-Time PCR (서양뒤영벌 야외개체군에서 Real-Time PCR을 이용한 Nosema ceranae의 검출)

  • Lee, Dae-Weon
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.270-274
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    • 2013
  • The bumblebee, Bombus terrestris, has played an important role as one of the alternative pollinators since the outbreak of honeybee collapse disorder. Recently, pathogens and parasites such as viruses, bacteria and mites, which affect the life span and fecundity of their host, have been discovered in B. terristris. In order to detect the microsporidian pathogen, Nosema spp. in the field populations of B. terristris, we collected adults and isolated their genomic DNA for diagnostic PCR. The PCR primers specific for Nosema spp. were newly designed and applied to gene amplification for cloning. Only small subunit ribosomal RNA (SSU rRNA) gene of N. ceranae was successfully amplified among examined genes and sequenced, which indicates that N. ceranae mainly infects the examined field population of B. terristris. To detect of SSU rRNA gene, two regions of SSU rRNA gene were selected by primary PCR analysis and further analyzed in quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis demonstrated that SSU rRNA of N. ceranae was detected at concentration as low as $0.85ng/{\mu}l$ genomic DNA. This result suggests that the detection via qRT-PCR can be applied for the rapid and sensitive diagnosis of N. ceranae infection in the field population as well as risk assessment of B. terristris.

NELL2 gene as regulator of cell cycle in neuron differentiation (신경세포 분화에서 세포주기 조절인자로서의 NELL2 유전자의 역할)

  • Joung, Mi Rim;Oh, Yeon Mi;Park, Woo Saeng;Park, Sang Kyu
    • Clinical and Experimental Pediatrics
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    • v.49 no.10
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    • pp.1100-1105
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    • 2006
  • Purpose : Because NELL2 expression is strictly restricted only in neurons in developing and post-differentiated neural tissues, it is thought to be involved in the neuronal differentiation during development and in the maintenance of neuronal physiology in the post-differentiated neurons. In this study, we examined whether NELL2 is involved in the regulation of cell cycle and apoptosis in the hippocampal neuroprogenitor HiB5 cells. Methods : Effects of NELL2 on the cultured HiB5 cell numbers, DNA fragmentation, and proteins involved in the regulation of the cell cycle were measured. Results : NELL2 induced a decrease in cell numbers and an increase in G1 phase arrest. Moreover, transfection of NELL2 resulted in an increase of DNA fragmentation that shows an evidence of apoptosis. Contents of proteins involved in the regulation of cell cycle were also changed by transfection of NELL2 expression vectors. Conclusion : This study suggests that NELL2 plays an important role in the regulation of cell cycle and apoptosis of neurons.

Evolutionary Relationship of Liobagrus mediadiposalis (Teleostei: Amblycipitidae) Populations in Korea Inferred from Cytochrome b DNA Sequences (한국 고유종인 자가사리(Liobagrus mediadiposalis) 지역개체군의 분자진화적 유연관계)

  • Kim, Maeng Jin;Han, Song-Hun;Yang, Hye-Young;Jo, Mi-Ran;Chung, Sang-Chul;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.18 no.4
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    • pp.329-338
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    • 2006
  • Phylogenetic relationships and DNA polymorphism among local populations of the Korean native catfish species, Liobagrus mediadiposalis, have been investigated based on mitochondrial cytochrome b DNA sequences. As a result, three genetically distinct groups of local populations were recognized based on the phylogenetic tree constructed. The first group was called "Nakdong-river group" that included the local populations from Geumho-river, Gyeongho-river, and Deokcheon-river; the second one was "Geum-river group"; the third one was represented as "Seomjin-river group" that included the samples from Seomjin-river, Dongjin-river and Geokum-do. The phylogeny also implied that the ancestral group of L. mediadiposalis have first evolved to Nakdong-river group, and later two local populations (Geum-river and Seomjinriver group) were diverged from the other lineage. DNA polymorphisms we observed were 4.4~4.7% between Geum-river group and Seomjin-river group, 5.1~5.5% between Seomjinriver group and Nakdong-river group, and 5.5~5.7% between Nakdong-river group and Geumriver group. These results indicated the long period of geographic isolation due to the river system in Korea caused such high degrees of DNA polymorphisms between local populations of L. mediadiposalis.

Serological and Histological Changes after Interferon Alfa Therapy in Children with Chronic Hepatitis B (소아 만성 B형 간염의 Interferon Alfa 치료 후 혈청학적, 조직학적 소견의 변화)

  • Ko, Jae-Sung;Chung, Ju-Young;Jang, Ja-Joon;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.3 no.1
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    • pp.56-62
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    • 2000
  • Purpose: The aim of this study was to evaluate the efficacy and histologic changes of interferon-alfa therapy on chronic hepatitis B virus infection in children. Patients and Methods: Thirty five children aged 3~16 years who were seropositive for HBV DNA, HBsAg and HBeAg were enrolled. Interferon-alfa 2a ($3.4\;MU/m^2$) were given for 6 months. Serologic markers of viral replication was evaluated 1 year after therapy. Post treatment liver biopsy was performed in 18 patients who showed serologic response. Results: Serum HBeAg and viral DNA became negative in 22 (63%) of treated children at 12 months after therapy. Serum aminotransferase levels normalized in all of the responders and HBsAg became negative in one responder. Horizontal transmission, serum aminotransferase levels more than twice normal, and active inflammation on liver biopsy were predictive factors for response to interferon therapy. Periportal piecemeal necrosis, lobular activity, portal inflammation, fibrosis, and total histologic activity index were reduced in responders. Conclusion: In children with chronic hepatitis B, interferon alfa promotes loss of viral replication and improves aminotransferase. Serologic response is associated with improvement in hepatic histology.

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Increased Apoptotic Efficacy of Decitabine in Combination with an NF-kappaB Inhibitor in Human Gastric Cancer AGS Cells (핵산합성 억제제인 decitabine과 NF-κB 활성 저해제인 PDTC의 병용 처리에 의한 인체 위암세포사멸 효과 증진)

  • Choe, Won Kyung;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1268-1276
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    • 2018
  • The cytidine analog decitabine (DEC) acts as a nucleic acid synthesis inhibitor, whereas ammonium pyrrolidine dithiocarbamate (PDTC) is an inhibitor of nuclear factor-${\kappa}B$. The aim of this study was to investigate the possible synergistic inhibitory effect of these two inhibitors on proliferation of human gastric cancer AGS cells. The inhibitory effect of PDTC on AGS cell proliferation was significantly increased by DEC in a concentration-dependent manner, and this inhibition was associated with cell cycle arrest at the G2/M phase and the induction of apoptosis. This induction of apoptosis by the co-treatment with PDTC and DEC was related to the induction of DNA damage, as assessed by H2AX phosphorylation. Further studies demonstrated that co-treatment with PDTC and DEC induced the disruption of mitochondrial membrane potential, increased the generation of intracellular reactive oxygen species (ROS) and the expression of pro-apoptotic Bax, and down-regulated the expression of anti-apoptotic Bcl-2, ultimately resulting in the release of cytochrome c from the mitochondria into the cytoplasm. Co-treatment with PDTC and DEC also activated caspase-8 and caspase-9, which are representative caspases of the extrinsic and intrinsic apoptosis pathways. Co-treatment also activated caspase-3, which was accompanied by proteolytic degradation of poly (ADP-ribose) polymerase. Taken together, these data clearly indicated that co-treatment with PDTC and DEC suppressed the proliferation of AGS cells by increasing DNA damage and activating the ROS-mediated extrinsic and intrinsic apoptosis pathways.

Taxonomic Review of Pristiophorus japonicus Complex (Pristiophoridae, Chondrichthyes) in the Northwest Pacific (북서태평양 톱상어(톱상어과, 연골어강)의 분류학적 재검토)

  • Myoung, Se Hun;Kim, Jin-Koo;Song, Choon Bok;Kai, Yoshiaki
    • Journal of Marine Life Science
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    • v.1 no.1
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    • pp.8-17
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    • 2016
  • The family Pristiophoridae belonging to the order Pristiophoriformes consists of about 8 species and 2 genera in the world. Among them, only one species (Pristiophorus japonicus) is distributed in northwestern Pacific. The family is characterized by having an elongated and flattened rostrum with laterally projecting a low of sharp teeth. In order to clarify variability of local populations of P. japonicus, we investigated 16 individuals collected from four locations (Korea, n=6; Miyagi, n=1; Kochi Pref., n=1; Okinawa, n=8) for morphological and molecular analyses. Morphological analysis showed that there may exist three morphotypes, of which A morphotype (Korea, Miyagi, Kochi Pref.) has a short snout (26.8% of TL), being most similar to syntypes of P. japonicus. On the other hand, B morphotype (Okinawa) has a long (31.7% of TL) and wider snout (snout width at nostrils 5.2 times pre-oral length) and, C morphotype (Okinawa) has a long (31.7% of TL) and narrower snout (6.3 times). Molecular analysis using mitochondrial DNA cytochrome b DNA sequences (386 base pairs) showed that the K2P distance between A and C morphotypes were 2.1~2.7%, but those within A morphotype were 0.0~0.5%. We identified A morphotype as P. japonicus, and designated B and C morphotypes as Pristiophorus sp. 1 and Pristiophorus sp. 2, respectively. Further studies are required to elucidate the taxonomic status of two morphotypes of the sawshark in Okinawa.

Effect of Glutathione S-Transferase Polymorphisms on the Antioxidant System (Glutathione S-Transferase 유전적 다형성이 항산화 체계에 미치는 영향)

  • Jeon, Gyeong-Im;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.6
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    • pp.708-719
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    • 2007
  • Glutathione S-transferase genotypes GSTT1, GSTM1 and GSTP1 were characterized in 104 healthy male and female subjects and compared with parameters of oxidative stress at the level of DNA and lipids, with antioxidant enzymes, and with plasma antioxidants in smokers and non.smokers. Of the 104 subjects studied, 57.4% were GSTT1 present and 47.6% were GSTM1 present. The GSTP1 polymorphisms a and b were represented as follows: a/a, 75.5%; a/b, 21.6%; b/b type, 2.9%. The GSTT1 null genotype was associated with decreased glutathione in erythrocytes and elevated lymphocytes DNA damage. GST-Px was higher in GSTT1 null compared with GSTT1 present type. The homozygous GSTP1 genotype was not associated with any antioxidant status or DNA damage. The difference in plasma ${\alpha}$-carotene and erythrocytes GSH-Px and GST activities between smokers and non-smokers was detected in the GSTT1 null genotype. Plasma ${\gamma}$-tocopherol and ${\beta}$-carotene decreased significantly in smokers having GSTM1 null genotype. When GSTT1 and GSTM1 were combined, plasma lycopene and erythrocyte GST were reduced in smokers in both null types of these genes. As for GSTP1 genotype, plasma ${\alpha}$-carotene and erythrocytes GSH-Px decreased significantly in smokers with GSTP1 b/b, while erythrocytes GSH-Px activities decreased in smokers with GSTP1 a/b. The different ${\beta}$-carotene level between smokers and non-smokers was seen with both GSTP1 a/a and a/b genotype. It seems that polymorphisms in the phase II metabolizing enzyme glutathione S-transferase may be important determinants of commonly measured biomarkers.

UNDERSTANDING OF EPIGENETICS AND DNA METHYLATION (후생유전학 (Epigenetics)과 DNA methylation의 이해)

  • Oh, Jung-Hwan;Kwon, Young-Dae;Yoon, Byung-Wook;Choi, Byung-Jun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.3
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    • pp.302-309
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    • 2008
  • Epigenetic is usually referring to heritable traits that do not involve changes to the underlying DNA sequence. DNA methylation is known to serve as cellular memory. and is one of the most important mechanism of epigenetic. DNA methylation is a covalent modification in which the target molecules for methylation in mammalian DNA are cytosine bases in CpG dinucleotides. The 5' position of cytosine is methylated in a reaction catalyzed by DNA methyltransferases; DNMTl, DNMT3a, and DNMT3b. There are two different regions in the context of DNA methylation: CpG poor regions and CpG islands. The intergenic and the intronic region is considered to be CpG poor, and CpG islands are discrete CpG-rich regions which are often found in promoter regions. Normally, CpG poor regions are usually methylated whereas CpG islands are generally hypomethylated. DNA methylation is involved in various biological processes such as tissue-specific gene expression, genomic imprinting, and X chromosome inactivation. In general. cancer cells are characterized by global genomic hypomethylation and focal hypermethylation of CpG islands, which are generally unmethylated in normal cells. Gene silencing by CpG hypermethylation at the promotors of tumor suppressor genes is probably the most common mechanism of tumor suppressor inactivation in cancer.