• Asian-Australasian Journal of Animal Sciences
• /
• 제26권4호
• /
• pp.588-595
• /
• 2013

Mesenchymal stem cells (MSCs) are often known to have a therapeutic potential in the cell-mediated repair for fatal or incurable diseases. In this study, canine umbilical cord MSCs (cUC-MSCs) were isolated from umbilical cord matrix (n = 3) and subjected to proliferative culture for 5 consecutive passages. The cells at each passage were characterized for multipotent MSC properties such as proliferation kinetics, expression patterns of MSC surface markers and self-renewal associated markers, and chondrogenic differentiation. In results, the proliferation of the cells as determined by the cumulative population doubling level was observed at its peak on passage 3 and stopped after passage 5, whereas cell doubling time dramatically increased after passage 4. Expression of MSC surface markers (CD44, CD54, CD61, CD80, CD90 and Flk-1), molecule (HMGA2) and pluripotent markers (sox2, nanog) associated with self-renewal was negatively correlated with the number of passages. However, MSC surface marker (CD105) and pluripotent marker (Oct3/4) decreased with increasing the number of subpassage. cUC-MSCs at passage 1 to 5 underwent chondrogenesis under specific culture conditions, but percentage of chondrogenic differentiation decreased with increasing the number of subpassage. Collectively, the present study suggested that sequential subpassage could affect multipotent properties of cUC-MSCs and needs to be addressed before clinical applications.

• 생약학회지
• /
• 제49권4호
• /
• pp.291-297
• /
• 2018

Coxsackievirus B3 (CVB3) is a very well-known causative agent for viral myocarditis and meningitis in human. However, the effective vaccine and therapeutic drug are not developed yet. CVB3 infection activates host cell AKT signaling. Inhibition of AKT signaling pathway may attenuate CVB3 replication and prevent CVB3-mediate viral myocarditis. In this study, we determined antiviral effect of the selected natural plant extract to develop a therapeutic drug for CVB3 treatment. We screened several chemically extracted natural compounds by using HeLa cell-based cell survival assay. Among them, Linum usitatissimum L. extract was selected for antiviral drug candidate. L. usitatissimum extract significantly decreased CVB3 replication and cell death in CVB3 infected HeLa cells with no cytotoxicity. CVB3 protease 2A induced eIF4G1 cleavage and viral capsid protein VP1 production were dramatically decreased by L. usitatissimum extract treatment. In addition, virus positive and negative strand genome amplification were significantly decreased by 1 mg/ml L. usitatissimum extract treatment. Especially, L. usitatissimum extract was associated with inhibition of AKT signal and maintain mTOR activity. In contrast, Atg12 and LC3 expression were not changed by L. usitatissimum extract treatment. In this study, the potential AKT signal inhibitor, L. usitatissimum extract, was significantly inhibited viral genome replication and protein production by inhibition of AKT signal. These results suggested that L. usitatissimum extract is a novel therapeutic agent for treatment of CVB3-mediated diseases.

• Fisheries and Aquatic Sciences
• /
• 제25권6호
• /
• pp.320-326
• /
• 2022

Most of the emerging diseases that threaten humans are caused by RNA viruses which are extremely mutable during evolution. The fish RNA virus, viral hemorrhagic septicemia virus (VHSV) can infect a broad range of aquatic animal hosts, but the transmissibility of VHSV to mammals has not been thoroughly investigated. Therefore, our study aimed to investigate the potential adverse effects of VHSV in mammals. Briefly, the survival of VHSV was determined using only minimum essential media (MEM-2) and mammalian SNU-1411 and hepa-1c1c7s cells at 15℃ and 37℃. Mice (Mus musculus, 27.3 ± 1.9 g) were intravenously injected with VHSV (2.37E+05 TCID₅₀·mice^-1) in triplicate. Clinical signs and survival rates were examined at 14 days post-challenge, and infection was confirmed in the surviving mice. The 50% tissue culture infective dose (TCID₅₀) and polymerase chain reaction analysis were used to determine viral titers and the infection rate, respectively. The titer of VHSV suspended in MEM-2 at 15℃ was reduced by only one log after 8 days, whereas the virus maintained at 37℃ was inactivated 8 days post-inoculation (dpi). There were no recognizable cytopathic effects in either SNU-1411 or hepa-1c1c7s cells inoculated with VHSV at 15℃ and 37℃. VHSV in those cell lines at 37℃ was rapidly decreased and eventually inactivated at 12 dpi, whereas virus at 15℃ remained at low concentrations without replication. In vivo experiment showed that there were no signs of disease, mortality, or infection in VHSV-infected mice. The results of this study indicate that it is highly unlikely that VHSV can infect mammals including humans.

• Pediatric Infection and Vaccine
• /
• 제17권2호
• /
• pp.130-136
• /
• 2010

목적 : 부산에서 급성호흡기 증상으로 외래 방문한 환자를 대상으로 원인 바이러스 및 역학을 연구하고자 하였다. 방법 : 2007년 1월부터 2008년 12월까지 3개 병원 소아청소년과 외래에 급성 호흡기 증상을 주소로 내원한 990명을 대상으로, 비강 세척법으로 검체를 채취하여 역전사중합효소연쇄반응을 시행하였다. 결과 : 총 검체 990건 중 351건(35.5%)이 검출되었으며, 남녀 비는 1.6대 1이었고, 5세 이하가 93.7%였다. hRV가 가장 흔한 원인으로 연중 202건(57.5%), 그 다음으로는 RSV로 10월부터 3월까지 57건(16.2%), ADV는 연중 37건(10.5%), INF는 12월부터 4월까지 21건(6%), hBoV는 1월부터 8월까지 15건(4.3%), PIV는 4월부터 7월까지 9건(2.6%), hCoV는 1월부터 7월까지 7건(2%), EV는 6월부터 9월에 3건(0.9%) 검출되었다. 결론 : 본 연구는 부산의 소아급성호흡기 질환을 예방하고 치료하는데 도움이 되고자 하며, 앞으로도 원인 바이러스에 대한 지속적인 연구가 필요하다.

• Tuberculosis and Respiratory Diseases
• /
• 제82권4호
• /
• pp.328-334
• /
• 2019

Background: Although the frequency of respiratory viral infection in patients with pulmonary acute respiratory distress syndrome (ARDS) is not uncommon, clinical significance of the condition remains to be further elucidated. The purpose of this study was to compare characteristics and outcomes of patients with pulmonary ARDS infected with influenza and other respiratory viruses. Methods: Clinical data of patients with pulmonary ARDS infected with respiratory viruses January 2014-June 2018 were reviewed. Respiratory viral infection was identified by multiplex reverse transcription-polymerase chain reaction (RT-PCR). Results: Among 126 patients who underwent multiplex RT-PCR, respiratory viral infection was identified in 46% (58/126): 28 patients with influenza and 30 patients with other respiratory viruses. There was no significant difference in baseline and clinical characteristics between patients with influenza and those with other respiratory viruses. The use of extracorporeal membrane oxygenation (ECMO) was more frequent in patients with influenza than in those with other respiratory viruses (32.1% vs 3.3%, p=0.006). Co-bacterial pathogens were more frequently isolated from respiratory samples of patients with pulmonary ARDS infected with influenza virus than those with other respiratory viruses. (53.6% vs 26.7%, p=0.036). There were no significant differences regarding clinical outcomes. In multivariate analysis, acute physiology and chronic health evaluation II was associated with 30-mortality (odds ratio, 1.158; 95% confidence interval, 1.022-1.312; p=0.022). Conclusion: Respiratory viral infection was not uncommon in patients with pulmonary ARDS. Influenza virus was most commonly identified and was associated with more co-bacterial infection and ECMO therapy.

• Childhood Kidney Diseases
• /
• 제19권2호
• /
• pp.176-179
• /
• 2015

Rituximab (RTX), a monoclonal antibody against the B-cell marker CD20, is commonly used as a treatment for antibody-mediated diseases or B-lymphocyte-mediated diseases. Destruction of B cells may reverse the disease course in many conditions; however, patients who are treated with RTX cannot respond appropriately to de novo infection due to lack of B lymphocytes. Here, we report one such case. A 7-year-old renal allograft recipient presented with severe anemia due to parvovirus infection after RTX treatment. The patient had focal segmental glomerulosclerosis and had received cadaveric kidney transplantation 6 months previously. She was treated with high-dose steroid for acute rejection and RTX for Epstein Barr Virus infection 3 months previously. At presentation, her hemoglobin level was 5.4 g/dL and leukocyte and platelet counts were normal. She had microcytic normochromic anemia and high viral load of parvovirus B19(70,578 copies/mL). Intravenous immunoglobulin ($200mg/kg{\cdot}d$) treatment controlled the progression of anemia and parvovirus infection. De novo parvovirus infection during the B lymphocyte-depletion period may have precipitated the severe anemia in this case. Close monitoring of infection is required after RTX therapy.

• Pediatric Infection and Vaccine
• /
• 제7권1호
• /
• pp.83-93
• /
• 2000

목 적 : Urabe AM-9 볼거리 백신주는 무균성 뇌막염의 발생 빈도가 높은 것으로 알려져 있다. 백신 접종 후 무균성 뇌막염을 일으키는 경우 Urabe AM-9 백신주의 hemagglutinin-neuraminidase(HN) 유전자의 염기서열 1081번의 G가 A로 치환되어 335번째 아미노산이 glutamic acid에서 lysine으로 바뀌게 됨에 따라 야생형의 mumps 바이러스와 동일한 작용을 나타내는 것으로 알려져 있다. 이에 국내에서 사용중인 Urabe AM-9 백신주의 안전성 여부를 조사하기 위해 일부 백신주와 야생 분리주의 HN 유전자의 염기서열 분석을 실시하였다. 방 법 : 국내에서 사용되고있는 Urabe AM-9주 백신 2종류와 1998~1999년에 볼거리 환자로부터 분리된 야생 분리주를 이용하여 RT-PCR 방법에 의해 HN 유전자를 증폭시킨 후 염기 및 아미노산 서열 분석을 실시하였다. 결 과 : 백신주와 야생 분리주 모두 HN 343, 1476, 1570번 위치에서의 염기 변이는 없었으나 1081번 위치 염기는 모두 야생주와 같은 Lysine/AAA form으로 나타났으며 변이주인 Glutamic acid/GAA와의 혼합 양상은 관찰되지 않았다. HN 1470번 위치의 염기는 백신주 중 계대 배양시 C에서 A로 치환되었으며, HN 1727번 위치 염기는 모두 A에서 G로 치환된 것으로 나타났다. 결 론 : 국내에서 사용중인 2종류의 Urabe AM-9 백신주와 야생 분리주 모두 1081번 위치의 염기가 야생주와 같은 무균성 뇌막염을 일으킬 생물학적 가능성이 있는 것으로 나타났으며, 국내에서도 상기 백신주를 사용한 후 무균성 뇌막염이 발생하고 있을 개연성이 매우 크다고 사료되며 직접적인 연관성에 대한 보완 연구를 위하여 정확하고 믿을만한 자료에 의한 실제 부작용 실태를 파악하여야 한다.

• 한국동물위생학회지
• /
• 제28권1호
• /
• pp.49-69
• /
• 2005

More than 30 years have elapsed since the first report of the International Committee on Taxonomy of Viruses (ICTV) was published in 1971. Since that publication, the ICTV recognizes about 1,550 virus species, but some 30,000 virus strains and isolates are being tracked by virologists in different fields of biology. The ICTV is the 'international court' of experts that rules on names and relationships of all virus, but only to the level of species. Virus taxonomy is changing rapidly, with changes ranging from the trivial(use of italics for species names) to profound reorganization driven by the explosion of sequence information. The universal system of viral taxonomy now accepts Linnean-like classification at the levels of order, family, subfamily, genus, and species. The suffix '-virales' identifies an order, Families are identified by the suffix '-viridae' subfamilies are identified by the suffix '-virinae', and genera are identified by the suffix '-virus'. The importance of distinguishing subspecies, strains, and isolates in vaccine development, diagnostics, etc. is recognized, but these lower levels are not formally classified by ICTV. This paper mainly introduces taxonomy and classification of animal viruses on the basis of the seventh report of the ICTV edited by Van Regenmortal et al. in 2000.

• BMB Reports
• /
• 제51권7호
• /
• pp.315-316
• /
• 2018

Genome engineering with clustered regularly interspaced short palindromic repeats (CRISPR) system can be used as a tool to correct pathological mutations or modulate gene expression levels associated with pathogenesis of human diseases. Owing to well-established local administration methods including intravitreal and subretinal injection, it is relatively easy to administer therapeutic genome engineering machinery to ocular tissues for treating retinal diseases. In this context, we have investigated the potential of in vivo genome engineering as a therapeutic approach in the form of ribonucleoprotein or CRISPR packaged in viral vectors. Major issues in therapeutic application of genome engineering include specificity and efficacy according to types of CRISPR system. In addition to previous platforms based on ribonucleoprotein and CRISPR-associated protein 9 derived from Campylobacter jejuni, we evaluated the therapeutic effects of a CRISPR RNA-guided endonuclease derived from Lachnospiraceae bacterium ND2006 (LbCpf1) in regulating pathological angiogenesis in an animal model of wet-type age-related macular degeneration. LbCpf1 targeting Vegfa or Hif1a effectively disrupted the expression of genes in ocular tissues, resulting in suppression of choroidal neovascularization. It was also notable that there were no significant off-target effects in vivo.

• Journal of Oral Medicine and Pain
• /
• 제24권2호
• /
• pp.181-187
• /
• 1999

The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of newer micromethodologies and increasing availability of immunological and molecular biological reagents. The outcome of researches in this field has already provided DNA probes and antibodies which can be used for diagnosing various kinds of diseases including inherited ones. This development can be also applied to diagnose diseases in oral and maxillofacial regions. Technological advances have yielded highly sensitive test methodologies so that low analyte concentration and small sample volume are no longer limiting factors. Therefore, saliva can be useful test fluid for an array of analytes. Salivary constituents of diagnostic significance include steroid hormones, antibodies, drugs, and tumor markers. Of the proteins present in saliva, viral-specific immunoglobulins are of the greatest diagnostic interest. The development of conjugates and antigens by recombinant DNA technique and peptide synthesis is necessary for clinical application. Several kits developed for the purpose of blood testing should be modified to permit their application to saliva. The final practical outcome of researches in diagnostic sciences will be various diagnostic agents which can be used for detection of bacteria and viruses, screening and diagnosis of diseases, genetic screening for forensic individual identification. For these purposes, collaboration researches and development between institutions and companies are essential.