• Journal of Microbiology
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• 제45권4호
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• pp.326-332
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• 2007

In order to understand the functional role of CPRl in Saccharomyces cerevisiae KNU5377 with regard to its multi-tolerance characteristics against high temperatures, inorganic acids, and oxidative stress conditions, whole cellular proteins were analyzed via liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). This procedure was followed by two-dimensional (2-D) gel electrophoresis. Under menadione stress conditions, the 23 upregulated proteins were clearly identified only in the wild- type strain of KNU5377. Among the proteins, Sodl1p Tsa1p, Ahp1, Cpr1p, Cpr3, Ssb2p, and Hsp12p were identified as components of antioxidant systems or protein-folding related systems. The CPR1 protein could not be completely detected in the $cpr1{\Delta}$ mutant of KNU5377 and the other upregulated proteins in the wild-type strain evidenced a clear correlation with the results of immunoblot analysis. Moreover, a reduction in growth patterns (about 50%) could be observed in the $cpr1{\Delta}$ mutant, as compared with that of the wild-type strain under mild MD stress conditions. These results indicate that the upregulation of CPR1 may contribute to tolerance against MD as an inducer of oxidative stress.

• 대한수의학회지
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• 제34권3호
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• pp.517-527
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• 1994

An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

• Journal of Microbiology
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• 제44권5호
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• pp.492-501
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• 2006

In this study, we attempted to characterize the physiological response to oxidative stress by heat shock in Saccharomyces cerevisiae KNU5377 (KNU5377) that ferments at a temperature of $40^{\circ}C$. The KNU5377 strain evidenced a very similar growth rate at $40^{\circ}C$ as was recorded under normal conditions. Unlike the laboratory strains of S. cerevisiae, the cell viability of KNU5377 was affected slightly under 2 hours of heat stress conditions at $43^{\circ}C$. KNU5377 evidenced a time-dependent increase in hydroperoxide levels, carbonyl contents, and malondialdehyde (MDA), which increased in the expression of a variety of cell rescue proteins containing Hsp104p, Ssap, Hsp30p, Sod1p, catalase, glutathione reductase, G6PDH, thioredoxin, thioredoxin peroxidase (Tsa1p), Adhp, Aldp, trehalose and glycogen at high temperature. Pma1/2p, Hsp90p and $H^+$-ATPase expression levels were reduced as the result of exposure to heat shock. With regard to cellular fatty acid composition, levels of unsaturated fatty acids (USFAs) were increased significantly at high temperatures ($43^{\circ}C$), and this was particularly true of oleic acid (C18:1). The results of this study indicated that oxidative stress as the result of heat shock may induce a more profound stimulation of trehalose, antioxidant enzymes, and heat shock proteins, as well as an increase in the USFAs ratios. This might contribute to cellular protective functions for the maintenance of cellular homeostasis, and may also contribute to membrane fluidity.

• 미생물학회지
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• 제48권2호
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• pp.102-108
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• 2012

2003년부터 2007년까지 서울지역 집단식중독 환자에 대하여 RT-PCR을 이용하여 노로바이러스를 검출하였으며, 총 4,685건 중 노로바이러스가 383건(8.2%)이 검출되었다. 식중독 발생 사례는 2003년부터 2007년까지 57사례가 발생하였다. 계절별 유행양상은 겨울철에 가장 많이 검출되었다. 섭취장소별 유행양상은 학교급식 중 위탁경영에 의한 발생이 32%로 나타났으며, 일반음식점에 의한 발생이 29%로 나타났다. 본 연구는 서울지역 노로바이러스성 식중독 관리를 위한 기초자료로 활용될 수 있을 것으로 기대된다.

• 식품산업과 영양
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• 제11권3호
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• pp.6-11
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• 2006

바이러스성 식중독은 장염을 일으키는 원인 병원체 중 노로바이러스에 의해 흔히 발생하며 이외에도 아스트로바이러스나 로타바이러스에 의한 집단 설사 사례가 국내에서 보고된 바 있다. 노로바이러스는 식중독과 관련하여 특히 오염된 식수와 굴 등 어패류의 생식을 통한 감염 사례가 많이 보고되어 있으나 사람 간 전파도 흔히 일어나는 전염력이 매우 높은 바이러스이다. 국내에서는 1999년 이후 보고가 되고 있으며 최근 집단 급식과 관련된 대형 식중독 사례들이 보고되면서 학교급식이 사회적인 이슈로 대두되고 있다. 2000년 이후 질병관리본부는 바이러스성 설사의 국내 발생현황을 파악하기 위하여 전국의 17개 시도보건환경 연구원과 노로바이러스를 포함한 4종의 바이러스성 장염원인 병원체에 대한 전국적인 실험실 감시체계를 운영한 결과 바이러스성 병원체가 확인된 사례의 약 18%에서 노로바이러스가 검출되었고, 집단설사 사례에서는 대부분 노로바이러스가 원인병원체로 확인되었다. 또한 노로바이러스의 조기 검출을 위해 질병관리본부는 2004년 중 노로바이러스 유전자 검출 kit를 자체적으로 제작하여 이를 전국의 시도 보건환경연구원을 연계한 감시체계에서 적극 활용함으로써 노로바이러스 집단설사사례의 조기 검출이 가능하게 되었고 지역내 노로바이러스 검출율을 높이는데 기여하였다. 국립보건연구원은 2003년과 2006년에 발생한 대규모 노로바이러스 식중독 사례 이외에도 산발적으로 지속적으로 발생하는 사례들을 조기에 탐지하고 국내에서 검출되는 설사바이러스 유전형 분포양상과 새로운 유전자형이나 변이주를 조기에 검출하고자 전국적인 노로바이러스 실험실 감시망을 강화하여 운영하고 있으며, 집단설사 발생시 각 사례의 연관성을 신속하게 분석할 수 있는 실시간 분자역학적 유전자 분석체계를 단계적으로 도입하고 있다. 실험실 감시체계 운영과 함께 집단 식중독 유발 병원체의 효율적인 관리를 위해 질병관리본부는 노로바이러스를 포함한 설사 유발 병원체를 신고대상 병원체로 지정(2006.06.12)하여 병원체 검출시 보고하도록 하고 관련 지침을 마련하였다. 노로바이러스가 지정전염병 병원체로 추가로 지정됨에 따라 집단 사례 및 실험실 감시사업을 통해 검출되는 병원체에 대한 보고가 강화되고 전파 방지와 2차 감염 사례 감소에도 기여할 수 있을 것으로 사료되며 전국의 실험실 감시망을 연결하는 국가 차원의 노로바이러스 실시간 분자역학적 분석체계 도입을 통해 노로바이러스 2차 감염을 줄이고 대규모 집단발병 및 유행의 조기 차단 효과를 가져올 수 있을 것이다.

• Journal of Ginseng Research
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• 제35권4호
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• pp.429-435
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• 2011

Korean red ginseng has been studied various biological activities such as immune, anti-oxidative, anti-microbial, and anticancer activities but antiviral mechanism needs further studies. In this study, we aimed to examine the antiviral effects of Korea red ginseng extract and ginsenosides on norovirus surrogate, including murine norovirus (MNV) and feline calicivirus (FCV). We evaluated the pre-, co-, and post-treatment effects of Korean red ginseng (KRG), ginsenosides $Rb_1$ and $Rg_1$. To measure the antiviral effect and cytotoxicity of KRG extract, and ginsenosides $Rb_1$ and $Rg_1$, we treated Crandell-Reese Feline Kidney for FCV or RAW264.7 cells for MNV with concentrations of 0, 5, 6.7, 10, 20 ug/mL total saponin. There was cytotoxic effect in the highest concentration 20 ug/mL of KRG extract so this concentration was excluded in this study. The FCV titer was significantly reduced to 0.23-0.83 $log_{10}$ 50% tissue culture infectious dose ($TCID_{50}$)/mL in groups pre-treated with red ginseng extract or ginsenosides. The titer of MNV was significantly reduced to 0.37-1.48 $log_{10}$ $TCID_{50}$/mL in groups pre-treated with red ginseng extract or ginsenosides. However, there was no observed antiviral effect in groups co-treated or post-treated with KRG and its constituents. Our data suggest that KRG extract has an antiviral effect against norovirus surrogates. The antiviral mechanisms of KRG and ginsenosides should be addressed in future studies.

• 한국동물위생학회지
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• 제13권2호
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• pp.162-183
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• 1990

From 1988 to 1989, 8 strains of canine parvovirus-2 (CPV-2) were isolated from the fecal specimens from the dogs that were clinically diagnosed as canine parvoviral enteritis in the veterinary hospitals located in the regions of Taejon and Chungbuk province. Studios on biological and physicochemical properties for the isolates were carried out. The results obtained by experiments are summarized as follows. 1. Among 62 fecal samples collected from the dogs with enteric diseases, 24 (38.7%) showed the haemagglutinating activity against porcine erythrocyte, ranging from 16 to 16, 384 of HA titers. 2. When 8 fecal specimens with high HA titer over 1, 000 were inocultated into CRFX cells, intranuclear inclusion bodies were obseverd in all of eight specimens, of w)lick three specimens showed cytoplasmic inclusions concurrently with the intranuclear inclusion bodies. 3. In study on species-specificity of haemagglutinating activity of the isolates, TJ-89-1 and TJ-89-2, it was found that the isolates revealed the highest haemagglutinating activity with porcine erythrocytes, showing the relatively lower haemagglutination titers with the erythrocytes from cat and rabbit. None of erythrocytes from other animals reacted with the isolates. 4. By the cross-haemagglutination inhibition test of the Isolates with reference viruses and sera, the Isolates were evidently identified as the strains of canine parvovirus-2. 5. In Physicochemical property test, it was evident that the isolates were stable in, lipid solvent, pH and heat treatment at $56^{\circ}C$ for 30 min. and contain DNA genome. 6. When seven puppies were inoculated intraorally with the isolate at HA titer of 8, 192, all of the puppies showed the symptoms of anorexia, vomiting, diarrhea and died at the 5th to 10th days post inoculation(pi). The fecal samples from all of the puppies revealed significantly high HA titers afterward the 5th days pi. Body temperature and the number of total leucocytes were slightly increased at the early stage of infection. but extremely decreased at the stage of collapse. HI titers of the sera started to increase at the 2nd to 3rd days pi reaching 512 to 1, 024 at the 4th to 5th day pi.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.207-217
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• 2007

The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.

• 한국동물위생학회지
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• 제32권1호
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• pp.11-18
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• 2009

Porcine epidemic diarrhea virus (PEDV), an enveloped single stranded RNA virus in the family Coronaviridae, causes acute viral enteric disease in piglets. Recently outbreaks of porcine epidemic diarrhea (PED) have been rare in Europe but frequent in Asia. In Korea, the increase of PED prevalence is showing specially in postweaning pigs. The purpose of this study was to investigate nucleotide sequence of nucleocapsid protein gene of PEDV field isolates from postweaning pigs in Korea and get more information about the viruses. A total of 15 postweaing pigs clinically suspected of PEDV infection by severe watery diarrhea and dehydration were used in this study. Viral RNA was extracted from small intestines and stools of the pigs. The N gene was amplified by nested RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. Three PEDVs were isolated from the suspected postweaning pigs. The N gene of three PEDV field isolates consisted of 483 nucleotides. These PEDV field isolates showed nucleotide sequence homology range from 99.6% to 95% with Chinese strains, from 99.8% to 95.2% with Korean strains, from 97.3% to 95.7% with Japanese strains and from 96.5% to 95.7% with Belgium and British strains. The encoded pritein shared range from 98.8% to 95.6% with Chinese strains, from 99.4% to 95% with Korean strains, from 97.5% to 96.3% with Japanese strains, from 95.6% to 95% with Belgium and British strains. By phylogenetic tree analysis based on nucleotide sequence, three PEDV field isolates were clustered into two groups which were Chinese isolate groups and other Korean isolate groups. These results indicated that some of PEDV field isolates prevailing in Korean postweaning pigs may be associated with those of Chinese strains and other Korean strains.

• 미생물학회지
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• 제47권4호
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• pp.323-327
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• 2011

2009년부터 2010년까지 서울지역에서 산발적으로 발생한 급성위장관염 환자에 대하여 효소면역측정법(ELISA)을 이용하여 로타바이러스를 검출하였다. 총 1,916건 중 로타바이러스는 354건(18.4%)이 검출되었다. RT-PCR을 이용하여 유전자형 분석을 실시한 결과 P6G[4] (35%), P8G[3] (28%), P8G[1] (24%), P4G[2] (10%), P8G[9] (3%)로 나타났다. 서울지역 지하수 70지점을 대상으로 효소면역측정법을 이용하여 로타바이러스를 조사한 결과 2 지점에서 양성으로 검출되었으며, RT-PCR을 이용하여 유전자형 분석을 실시한 결과 P8G[3]로 나타났다. 이러한 연구결과는 서울지역 로타바이러스 질환의 예방을 위한 기초자료로 활용될 것으로 기대된다.