• 한국식품위생안전성학회지
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• 제39권3호
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• pp.231-238
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• 2024

국내에서 V. parahaemolyticus로 인한 식중독 사고가 지속적으로 보고되고 있으며, 최근 국내 수산물 판매량 및 수산물 양식에 사용되는 항생제 판매량은 증가하는 추세이다. 따라서 본 연구는 국내에 유통되는 수산물에서 분리한 V. parahaemolyticus의 분포, 항생제 감수성, 유전적 특성 및 유전학적 통계를 조사하였다. 79건의 유통 수산물로부터 47건(59.5%)에서 V. parahaemolyticus가 분리되었다. 항생제 내성 양상의 경우, 총 47균주의 분리 균주에서는 ampicillin에 2균주(4.3%)가 내성을 보였으며, 이외 균주는 모든 항생제에 대해 감수성을 보였다. 항생제 내성 유전자의 경우, 모든 균주(100%)로부터 bla_CAR family gene, tet(35), catC가 확인되었으며, 1균주(2.1%)에서는 fos가 확인되었다. 병원성 유전자 여부의 경우, 모든 분리 균주에서 tdh, trh 유전자는 확인되지 않았으나, T3SS1은 모든 균주(100%), T3SS2는 1균주(2.1%)에서 확인되었다. MLST의 경우, 17균주로부터 15가지의 ST가 확인되었으며, ST 658가 3균주, 이외 14가지 ST는 1균주씩 확인되었다. 확인된 ST는 대부분 중국, 태국 등의 환경 분리주로 확인되었으며, ST 396, ST 3042는 중국 임상 분리주로부터 확인되었다. 이로써, 최근 국내에 수산물과 관련한 식중독, 유통량, 항생제 판매량 등의 추세에 따른 위험성에 V. parahaemolyticus에 대한 지속적인 연구가 필요할 것으로 사료되며, 본 연구는 그에 대한 도움이 될 것이라 사료된다.

• Journal of Ginseng Research
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• 제39권3호
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• pp.213-220
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• 2015

Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.

• 약학회지
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• 제51권1호
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• pp.75-82
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• 2007

Reactive oxygen species (ROS) are produced in the metabolic process of oxygen in cells. The superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in cells systemize the antioxidant enzymes to control the oxidative stress. Genistein is one of the isoflavonoids, and its role in controlling cellular oxidative stress is presently the active issue at question. In this study; we analyzed genistein-induced survival rates of the CHO-K1 cells, activities of antioxidant enzymes, ROS levels, and expression levels of antioxidant enzyme genes in order to investigate the effect of genistein on cellular ROS production and antioxidative systems in CHO-K1 cells. As results, the survival rate of cells was decreased as the dose of genistein increases (12.5${\sim}$200 ${\mu}$M). Genistein increased cellular ROS levels, while it reduced total SOD activities and the expression of CuZnSOD. In conclusion, we suggest that genistein may induce oxidative stress via down-regulation of SOD.

• 미생물학회지
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• 제21권2호
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• pp.61-70
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• 1983

Synchronized cat kidney cells chronically infected with feline leukemia virus (FeLV) were used to study virus production, the synthesis of group specific antigen (gag) and envelope (env) proteins, the expression of env protein on the cell surface during the cell cycle, and the stability of viral RNA. As detecting method, we developed the radioimmunoassay (RIA) system using beta-emission of $^{131}I$ and demonstrated the validity of this system by comparison with routine RIA system using gamma-emission of $^{125}I$. The produced virus was analysed by developed RIA interval was determined by measuring reverse transcriptase activity. The results show that infected cells produce the complete virus particle containing products of gag, env and pol genes of FeLV, and maximum virus production occurs during mitosis of synchronized cells. Labeling of the cell surface of synchronized cells with $^{131}I$ shows that the amount of $gp70^{env}$ on the cell surface parallels cellular gorwth. Therefore, the cell cycle-dependent release of virus is not petition RIA of synchronized cells with $^{131}I$ labeled viral proteins synthesis during the cell cycle. The rate of synthesis of gag protein shows three peaks, corresponding to the $G_1,\;late\;S\;and\;late\;G_2$ phases of cell cycle. But the rate of synthesis of env protein dose not change, suggesting that in these cells the synthesis of these two gene products in controlled seperately. In Actionomycin D treated cells, the synthesis of viral proteins decreased sharply from 8 hours after treatment, and the late S and $G_2$ peaks of gag protein synthesis were disappeared. This shows the stability of viral RNA for about 6 hours in the absence of continuing viral RNA synthesis.

• Journal of Animal Science and Technology
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• 제58권11호
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• pp.39.1-39.12
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• 2016

Background: The current study investigates the anti-stress effects of clove (Eugenia caryophyllus) extracts (0, 200, 400, and 600 mg/kg) on serum antioxidant biomarkers, immune response, immunological organ growth index, and expression levels of acute phase proteins (APPs); ovotransferrin (OVT), ceruloplasmin (CP), ceruloplasmin (AGP), C-reactive protein (CRP), and serum amyloid-A (SAA) mRNA in the immunological organs of 63-d-old male black-meated Silkie fowls subjected to 21 d chronic heat stress at $35{\pm}2^{\circ}C$. Results: The results demonstrated that clove extract supplementation in the diet of Silkie fowls subjected to elevated temperature (ET) improve growth performance, immune responses, and suppressed the activities of glutathion peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), and thioredoxin reductase (TXNRD); reduced serum malonaldehyde (MDA) and glutathione (GSH) concentrations when compared with fowls raised under thermoneutral condition (TC). Upon chronic heat stress and supplementation of clove extracts, the Silkie fowls showed a linear increase in GSH-Px, SOD, CAT, and TXNRD activities (P = 0.01) compared with fowls fed diets without clove extract. ET decreased (P < 0.05) the growth index of the liver, spleen, bursa of Fabricius and thymus. However, the growth index of the liver, spleen, bursa of Fabricius and thymus increased significantly (P < 0.05) which corresponded to an increase in clove supplemented levels. The expression of OVT, CP, AGP, CRP, and SAA mRNA in the liver, spleen, bursa of Fabricius and thymus were elevated (P < 0.01) by ET compared with those maintained at TC. Nevertheless, clove mitigates heat stress-induced overexpression of OVT, CP, AGP, CRP and SAA mRNA in the immune organs of fowls fed 400 mg clove/kg compared to other groups. Conclusions: The results showed that clove extracts supplementation decreased oxidative stress in the heat-stressed black-meated fowls by alleviating negative effects of heat stress via improvement in growth performance, antioxidant defense mechanisms, immunity, and regulate the expression of acute phase genes in the liver and immunological organs.

• 한국임상수의학회지
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• 제23권4권
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• pp.405-410
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• 2006

피부사상균증이 있는 개와 고양이에서 분리한 9주의 Microsporum canis와 5주의 Microsporum gypseum에서 ribosomal DNA를 추출하여 internal transcribed spacer 1 (ITS1) gene을 PCA로 증폭한 후 sequencing을 실시하여, 각 사상균의 계통학적 관계를 조사하였다. M canis 분리주 9주의 ITS1 gene의 nucleotide sequence는 100% 일치하였으며 M gypseum 분리주 5주의 nucleotide sequence도 100% 일치하였다. M canis 분리주 9주의 계통분석 결과 미국, 일본, 호주 및 유럽에서 분리된 M canis와 같은 cluster에 속하였으며 다른 Microsporum spp와는 유전적으로 다른 cluster를 형성하였다. 그러나 M canis와 M distortum, M equinum, M ferrugineum은 유전적으로 매우 가까운 위치에 있었다. M gypseum 분리주는 M canis와는 다른 cluster를 형성하였다. ITS1 gene의 분자생물학적 분석은 Microsporum spp를 확인하고 그들의 유전학적 관계를 이해하는 유용한 정보를 제공하는 것으로 생각된다.

• Molecules and Cells
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• 제19권1호
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• pp.97-103
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• 2005

The role of residue C323 in catalysis by human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) was examined by substituting Arg, Gly, Leu, Met, or Tyr at C323 by cassette mutagenesis using synthetic human GDH isozyme genes. As a result, the $K_m$ of the enzyme for NADH and ${\alpha}-ketoglutarate$ increased up to 1.6-fold and 1.1-fold, respectively. It seems likely that C323 is not responsible for substrate-binding or coenzyme-binding. The efficiency ($k_{cat}/K_m$) of the mutant enzymes was only 11-14% of that of the wild-type isozymes, mainly due to a decrease in $k_{cat}$ values. There was a linear relationship between incorporation of [$^{14}C$]p-chloromercuribenzoic acid and loss of enzyme activity that extrapolated to a stoichiometry of one mol of [$^{14}C$] incorporated per mol of monomer for wild type hGDHs. No incorporation of [$^{14}C$]p-chloromercuribenzoic acid was observed with the C323 mutants. ADP and GTP had no effect on the binding of p-chloromercuribenzoic acid, suggesting that C323 is not directly involved in allosteric regulation. There were no differences between the two hGDH isozymes in sensitivities to mutagenesis at C323. Our results suggest that C323 plays an important role in catalysis by human GDH isozymes.

• 미생물학회지
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• 제33권2호
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• pp.92-96
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• 1997

Pseudomonas sp. strain DJ77로부터 클로닝한 catechol 분해와 관련된 phnDEFG 유전자들이 존재하는 pHENX7에서 phnF 유전자의 염기서열을 밝혔다. Extradiol dioxygenase 유전자인 phnE와, 2-hydroxymuconic semialdehyde dehydrogenase를 생산하는 phnG 유전자 사이에 존재하는 유전자 phnF는 432 bps로 된 하나의 open reading frame(ORF)으로 존재하였고, 여기서 유추한 아미노산은 143개로 분자량 13,859 dalton의 polypeptide를 만들어 내고 있다. phnF 유전자는 Sphingomonas sp. strain HV3 catE 유전자 부위와 sphingomonas yanoikuyae B1의 xylE와 xylG 사이에 존재하는 ORF 부위의 염기서열과 각각 99%, 68.6%의 상동성을 가지고 있었다. 또한 PhnF 단백질의 아미노산서열은 citrobacter freundii DSM30040의 orfY 부위의 아미노산서열과 62.3%의 상동성이 있었다.

• Biomolecules & Therapeutics
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• 제31권1호
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• pp.82-88
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• 2023

Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni²⁺-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with K_d values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a k_cat value of 0.038 ± 0.002 min^-1 and a K_m value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a k_cat value of 0.135 ± 0.007 min^-1 and a K_m value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

• 한국동물위생학회지
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• 제45권4호
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• pp.305-316
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• 2022

Bordetella (B.) bronchiseptica, Mycoplasma (M.) felis, and Chlamydia (C.) felis are considered as main bacterial pathogens of feline upper respiratory tract disease (URTD). In this study, a new triplex quantitative real-time polymerase chain reaction (tqPCR) assay was developed for the rapid and differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with the detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 1%. Based on the diagnostic results of the assay using 94 clinical samples obtained from cats with URTD signs, prevalence of B. bronchiseptica, M. felis, or C. felis was 10.6%, 36.2%, or 6.4%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported monoplex qPCR assays. The dual infection rates for B. bronchiseptica and M. felis or M. felis and C. felis was 2.1% or 3.2%, respectively. These results indicated that M. felis has been widely spread, and its co-infection with B. bronchiseptica or M. felis has been frequently occurred in Korean cat population. The developed tqPCR assay will serve as a promising tool for etiological and epidemiological studies of these three bacterial pathogens and the prevalence data obtained in this study will contribute to expanding knowledge about the epidemiology of feline URTD in Korea.