• Journal of Microbiology
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• 제45권3호
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• pp.187-192
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• 2007

REOviruses (Respiratory Enteric Orphan viruses) are ubiquitous, non-enveloped viruses containing 10 segments of double-stranded RNA (dsRNA) as their genome. They are common isolates of the respiratory and gastrointestinal tract of humans but are not associated with severe disease and are therefore considered relatively benign. An intriguing characteristic of reovirus is its innate oncolytic potential, which is linked to the transformed state of the cell. When immortalized cells are transfected in vitro with activated oncogenes such as Ras, Sos, v-erbB, or c-myc, they became susceptible to reovirus infection and subsequent cellular lysis, indicating that oncogene signaling pathways are exploited by reovirus. This observation has led to the use of the virus in clinical trials as an anti-cancer agent against oncogenic tumors. In addition to the exploitation of oncogene signaling, reovirus may further utilize host immune responses to enhance its antitumor activity in vivo due to its innate interferon induction ability. Reovirus is, however, not entirely benign to immunocompromised animal models. Reovirus causes so-called "black feet syndrome" in immunodeficient mice and can also harm neonatal animals. Because cancer patients often undergo immunosuppression due to heavy chemo/radiation-treatments or advanced tumor progression, this pathogenic response may be a hurdle in virus-based anticancer therapies. However, a genetically attenuated reovirus variant derived from persistent reovirus infection of cells in vitro is able to exert potent anti-tumor activity with significantly reduced viral pathogenesis in immunocompromised animals. Importantly, in this instance the attenuated, reovirus maintains its oncolytic potential while significantly reducing viral pathogenesis in vivo.

• 한국식품영양과학회지
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• 제42권10호
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• pp.1567-1575
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• 2013

본 연구에서는 유황오리의 우수성을 확인하기 위하여 유황오리, 일반오리, 닭을 열수추출 하여 유황오리의 대장염 염증에 대한 억제 효과를 알아보고자 하였다. Balb/c 수컷 마우스에 2.5% DSS로 궤양성 대장염을 유도하였고 이에 대한 대장염의 염증 억제효과를 검토하였다. 모든 실험군 중에서 정상군의 대장 길이가 가장 길었고, 대조군의 대장 길이가 가장 짧았다. 유황오리 고농도군, 저농도군의 대장의 길이는 일반오리 고농도군 저농도군의 대장 길이보다 길었으며, 닭고농도군, 저농도군의 대장의 길이는 일반오리 추출물보다 짧았다. 유황오리 저농도군에서 대장 길이가 시료처리군 중에서 가장 길었고, 대장염 증상의 개선 효과를 나타내었다. 대장의 조직학적 관찰에서 대장의 길이의 축소 정도가 증가함에 따라 대장조직의 점막층의 선와부의 손실이 증가되었다. 대장염 염증 초기에 증가하는 염증성 biomarker인 혈청 IL-$1{\beta}$, IL-6, TNF-${\alpha}$, IL-17A, IL-12의 생성은 유황오리 저농도군에서만 대조군보다 유의적으로 감소하였고, 정상군과 비슷한 수준을 보였다(P<0.05). 대장염을 일으킨 대장조직에서의 염증성인자인 IL-6, TNF-${\alpha}$, COX-2, iNOS의 mRNA 발현에서도 유황오리 추출액이 일반오리 추출액보다 더 많이 감소하였고, 유황오리 저농도군에서 가장 많이 감소되었다. 이상의 결과로 DSS로 대장염이 유도된 마우스에서 유황오리 추출액이 일반오리와 닭 추출액보다 더 높은 항염증 효과를 나타내었고 처리 농도에 따른 차이가 있었으며, 유황오리 추출액을 1 mL/kg으로 투여하였을 때 대장의 염증을 억제하는 효과가 가장 우수하였다.

• Molecular & Cellular Toxicology
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• 제5권3호
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• pp.207-215
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• 2009

Despite wide therapeutic use of CpG ODN against infection, allergy and cancer, the safety and toxicity of CpG ODNs were poorly delineated. Thus, we investigated whether optimal dosing of CpG ODN would affect immunotoxicological parameters in NZB/NZW F1 mice. Comparisons were made among control, non-CpG ODN and mouse CpG ODN ($10{\mu}g$)-treated groups for 4 weeks. To gauge the immunotoxicity of CpG ODNs, we measured nonspecific parameters, degree of lupus nephritis, proteinuria, or autoantibody, and cytokine expression in mRNA level of lymphocytes. We found that there were no significant differences among groups in nonspecific immunotoxicological profiles and in evaluation profiles of glomerulonephritis. However, titer of anti-dsDNA and anti-cardiolipin antibodies in mouse CpG ODN group rose three or eight-fold higher than in control group. Collectively, CpG ODN might be clinically less immunotoxic in terms of clinical profiles in lupus-prone NZB/NZW F1 mice, in spite of high autoantibody titer in CpG ODN treated groups.

• The Plant Pathology Journal
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• 제19권1호
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• pp.30-33
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• 2003

The chestnut blight fungus, Cryphonectria parasitica, and its hypovirus aye a useful model system in the study of the mechanisms of hypoviral infection and its consequences, such as a biological control of fungal pathogens. Strains containing the double-stranded (ds) RNA viruses Cryphonectria hypovirus 1 show characteristic symptoms of hypovirulence and display hypovirulence-associated changes, such as reduced pigmentation, sporulation, laccase production, and oxalate accumulation. Interestingly, symptoms caused by hypoviral infection appear to be the result of aberrant expression of a number of specific genes in the hypovirulent strain. Several viral regulated fungal genes are identified as cutinase gene, Lac1, which encodes an extracellular laccase, Crp, which encodes an abundant tissue-specific cell-surface hydrophobin that mediates physical strength, and Mf2/1 and Mf2/2, which encode pheromone genes involved in poor sporulation in the presence of hypo-virus. Since the phenotypic changes in the fungal host are pleiotropic, although coordinated and specific, it has been suggested that the hypovirus disturbs one or several regulatory pathways (Nuss,1996). Accordingly, several studies have shown the implementation of a signal transduction pathway during viral symptom development. Although further studies are required, hypovirulence and its associated symptom development due to the hypoviral regulation of a fungal hetero-trimeric G-protein have been suggested. In addition, recent studies have shown the presence of a novel protein kinase gene cppk1 and its transcriptional upregulation by hypovirus. In this review, the presence of important components in signal transduction pathway, their putative biological function, and viral-specific regulation will be addressed.

• Molecules and Cells
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• 제43권8호
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• pp.671-685
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• 2020

The regulator of calcineurin (RCAN) was first reported as a novel gene called DSCR1, encoded in a region termed the Down syndrome critical region (DSCR) of human chromosome 21. Genome sequence comparisons across species using bioinformatics revealed three members of the RCAN gene family, RCAN1, RCAN2, and RCAN3, present in most jawed vertebrates, with one member observed in most invertebrates and fungi. RCAN is most highly expressed in brain and striated muscles, but expression has been reported in many other tissues, as well, including the heart and kidneys. Expression levels of RCAN homologs are responsive to external stressors such as reactive oxygen species, Ca²⁺, amyloid β, and hormonal changes and upregulated in pathological conditions, including Alzheimer's disease, cardiac hypertrophy, diabetes, and degenerative neuropathy. RCAN binding to calcineurin, a Ca²⁺/calmodulin-dependent phosphatase, inhibits calcineurin activity, thereby regulating different physiological events via dephosphorylation of important substrates. Novel functions of RCANs have recently emerged, indicating involvement in mitochondria homeostasis, RNA binding, circadian rhythms, obesity, and thermogenesis, some of which are calcineurin-independent. These developments suggest that besides significant contributions to DS pathologies and calcineurin regulation, RCAN is an important participant across physiological systems, suggesting it as a favorable therapeutic target.

• Journal of Microbiology and Biotechnology
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• 제30권12호
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• pp.1843-1853
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• 2020

COVID-19, caused by the novel coronavirus SARS-CoV-2, has spread globally and caused serious social and economic problems. The WHO has declared this outbreak a pandemic. Currently, there are no approved vaccines or antiviral drugs that prevent SARS-CoV-2 infection. Drugs already approved for clinical use would be ideal candidates for rapid development as COVID-19 treatments. In this work, we screened 1,473 FDA-approved drugs to identify inhibitors of SARS-CoV-2 infection using cell-based assays. The antiviral activity of each compound was measured based on the immunofluorescent staining of infected cells using anti-dsRNA antibody. Twenty-nine drugs among those tested showed antiviral activity against SARS-CoV-2. We report this new list of inhibitors to quickly provide basic information for consideration in developing potential therapies.

• 한국동물학회지
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• 제27권2호
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• pp.103-116
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• 1984

유전자 구조 및 유전정보 흐름의 차이로 인하여 고등생물의 유전자를 미생물에 직접 cloning하면 원하는 유전자 산물을 얻지 못하는 경우가 많다. 이것을 극복하기 위해서는 화학적인 방법으로 유전자를 합성하든지, 또는 역제효소를 사용하여 고등생물의 mRNA로부터 유전자를 합성하여 cloning하는 방법을 사용한다. 본 연구에서는 oligo(dT)-cellulose column 방법으로 순수분리한 plasmid pBR322의 Pst I site에 cloning하였다. 우선 AMV reverse transcriptase로 primary cDNA를 합성하고, 알칼리를 처리하여 주형 RNA를 제거했다. 이번에는 이 primary cDNA를 주형으로 Klenow enzyme과 reverse transcriptase를 차례로 처리하여 double stranded DNA를 합성하고, 이 때 5' end 근처에 형성되는 hairpin loop을 Sl nuclease로 제거했다. Terminal deoxynucleotidyl transferase를 사용하여, 합성된 dsDNA에는 poly(dC) track을, Pst I endonuclease를 처리한 plasmid DNA에서는 poly(dG) track을 각각 붙인다음 이들을 서로 annealing시키고 E. coli에 transformation시켜서 크기가 큰 plasmid를 갖는 clone을 cracking 방법으로 일처 선별하였다. 이렇게 선별된 clone을 in 냐셔 hybridization 방법으로 조사하여 globin DNA가 들어간 colony를 이차 선별하고 여러 restriction enzyme으로 잘라보아 globin DNA가 cloning된 것을 확인하였다. 토끼 hemoglobin으로 immunize한 rat (Wistar)에서 뽑은 제일차 혈청과 염소에서 뽑은 제이차 혈청의 antibody를 사용한 radioimmunoassay방법으로, cloning된 globin gene이 대장균내에서 발현되는 지의 여부를 살펴 보았는데, 박테리아의 $\\beta$-lactamase와 토끼의 globin이 결합된 chimeric protein이 대장균 내에서 다량 합성되며, 이 단백질은 토끼 hemoglobin의 antigenic determinant를 가지고 있음을 알 수 있었다.

• 해양환경안전학회:학술대회논문집
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• 해양환경안전학회 2007년도 추계학술발표회
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• pp.185-185
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• 2007

20세기에 들어 산업, 군사 및 다양한 목적으로 비인화성 용매인 PCE와 TCE의 사용량이 증대하였다. 주의를 필요로 하는 물질임에도 불구하고 부주의한 사용과 보관으로 인해 토양, 퇴적토, 지하수에 심각하게 오염되었다. High-chlorinated ethenes은 호기성 박테리아의 oxygenation에 의해 분해되지 않는다. PEC및 TCE의 완전한 탈염소화는 혐기성조건에서만 관찰되어지며, 지난 10연년간의 연구에 의해서 탈염소화 혐기성 미생물의 수의 보고는 증가되었다. 혐기성 조건에서 탈염소화 미생물에 의해 PCE와 TCE는 less-chlorinated ethenes 또는 무해한 ethene으로 전환이 가능하다. 본 연구는 lactate를 electron donor로 이용해 PCE에서 ethene까지 완전히 탈염소화하는 혐기성 배양을 수행했다. PCE로 오염된 퇴적토 시료로부터 혐기성 미생물 배양을 성공했다. PCE가 ethene까지 완전히 분해되는 것이 관찰되었다. 추가적으로 혐기성 미생물 배양액에서 1,2-cis-dichloroethene (cis-DCE)와 vinyl chloride (VC)의 축적이 일어남을 관찰하였다. 혐기성 미생물 배양액에서 Dehalococcoides 16S rRNA gene sequences에 특이적으로 반응하는 primer를 이용한 DGGE를 통해 미생물 군집을 분석하였다. 결론적으로, 우리의 연구에서 PCE를 감소시키는 배양액을 배양했으며, 이 배양엑에는 Dehalococcoides sp. 존재하는 것을 확인하였다.

• The Plant Pathology Journal
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• 제16권5호
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• pp.269-279
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• 2000

Alfalfa mosaic alfamoviruses(AIMV) were isolated from infected potato (Solanum tuberosum) and azuki bean (Paseolus angularis) in Korea. Two AIMV isolated from potatoes were named as strain KR (AIMV-KR1 and KR2) and AIMV isolated from azuki bean was named as strain Az (AIMV-Az). Each isolated AIMV strain was characterized by using their host ranges, symptom developments, serological relations and nucleotide sequence analysis of coat protein (CP) gene. Strains KR1, KR2, and Az were readily transmitted to 20 of 22 inoculated plant species including bean, cowpea, tomato, tobacco, and potato. AIMV-KR1 and KR2 produced the typical symptoms like chlorotic or necrotic spots in Chenopodium quinoa and Solanum tuberosum cv. Superior. AIMV-Az caused bright yellow mosaic symptom and leaf malformation in Nicotiana glauca, which were different from the common mosaic symptom caused by AIMV-KR1 and KR2. Electron microscope observation of purified virus showed bacilliform virions containing a single-stranded plus-strand RNAs of 3.6, 2.6, 2.0 and 0.9 kbp in length, respectively, similar in size and appearance to those of Alfamovirus. In SDS-PAGE, the coat protein of the two viruses formed a consistent band that estimated to be about 24kDa. The CP genes of the AIMV strains, KR1, KR2, and Az have been amplified by RT-PCR using the specific primers designed to amplify CP gene from viral RNA-3, cloned and sequenced. Computer aided analysis of the amplified cDNA fragment sequence revealed the presence of a single open reading frame capable of encoding 221 amino acids. The nucleotide and peptide sequence of viral CP gene showed that strain KR1, KR2, and Az shared highest nucleotide sequence identities with AIMV strain 425-M at 97.7%, 98.2%, and 97.2%, respectively. CP gene sequences of two strains were almost identical compared with each other. Altogether, physical, serological, biological and molecular properties of the purified virus.

• Molecules and Cells
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• 제26권5호
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• pp.496-502
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• 2008

Cryparin, encoded as a single copy gene (Crp) of the chestnut blight fungus Cryphonectria parasitica, is the most abundant protein produced by this fungus. However, its accumulation is decreased remarkably in C. parastica strains containing the double-stranded (ds) RNA virus Cryphonectria hypovirus 1. To characterize the transcriptional regulatory element(s) for strong expression and viral regulation, promoter analysis was conducted. Serial deletion of the Crp promoter region resulted in a step-wise decrease in promoter activity, indicating a localized distribution of genetic elements in the cryparin promoter. Promoter analysis indicated two positive and a repressive cis-acting elements. Among them, the promoter region between nt -1,282 and -907 appeared to be necessary for hypoviral-mediated down-regulation. An electrophoretic mobility shift assay (EMSA) on the corresponding promoter region (-1,282/-907) indicated two regions at (-1,257/-1,158) and (-1,107/-1,008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.