• Title/Summary/Keyword: viruses

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A Survey on the Workplace Environment and Personal Protective Equipment of Poultry Farmers (양계 농업인의 작업장 환경 및 개인보호구 착용 실태조사)

  • Kim, Insoo;Kim, Kyung-Ran;Lee, Kyung-Suk;Chae, Hye-Seon;Kim, Sungwoo
    • Journal of Environmental Health Sciences
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    • v.40 no.6
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    • pp.454-468
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    • 2014
  • Objectives: This study was conducted to investigate the actual condition of the farm work environment and personal protective equipment as part of the effort to improve livestock work for the safety and health of poultry farmers and provide basic data for establishing plans to improve and develop personal protective equipment. Methods: For this purpose, a questionnaire survey on general information about stables, the poultry work environment, accidents, the wearing of work clothes and personal protective equipment, and the level of awareness related to personal protective equipment was conducted among 148 poultry farmers. Results: As a result, it was found that poultry workplace environment was exposed to such risks as fine dusts; organic dusts; poisonous gases; odorous substances; chicken excrement; contact with chickens, bacteria or viruses; and accidents related to machine operation. Thirteen percent of respondents suffered severe respiratory diseases, and the most frequently injured sites due to accidents were the hands (25.7%), knees (23.8%), arms (17.3%), and head (10.9%). The most frequent type of accident was collisions between the body and obstacles or machinery during movement (36.4%), followed by erroneous machine operation such as feeders and electric shocks (8.5%). Regarding the wearing of work clothes and personal protective equipment, 51.7% of the respondents wore worn-out clothing or everyday clothes, whereas only 32.0% wore work clothes. The percentage of farmers who wore proper protective equipment for the work environment during poultry work was 48.4%. The most frequently used type of protective equipment was boots (38.9%), followed by mask (36.7%), gloves (36.3%), appropriate work clothes (22.6%), quarantine clothes (17.6%), helmets (13.4%), and goggles (12.6%). The rate of wearing goggles was low because they were considered inconvenient and lowered work efficiency. Furthermore, they purchased everyday products available on the market for their personal protective equipment which were not appropriate for maintaining safety in an actual harmful environment and its consequent risks. As a result of the survey of the awareness level related to personal protective equipment, their levels of awareness of accidents and attitude proved to be average or higher, but the practice of wearing protective equipment and the level of knowledge and management of personal protective equipment were lower. Conclusion: This survey found that the wearing status of personal protective equipment among poultry farmers was insufficient even though they were exposed to risks. Most respondents were aware of the necessity of wearing personal protective equipment and of the potential for accidents, but they did not wear proper protective equipment. Their wearing rate was low due to a lack of knowledge about protective equipment, as well as the inconvenience of wearing it. Therefore there is a need to improve and develop specialized personal protective equipment for respiration, hands, and eyes, as well as work clothes that can protect farmers from major harmful matter that is generated in the poultry workplace. Based on the results of this investigation, we will conduct further studies on the required performance and design directions of personal protective equipment while collecting more objective data through field-oriented assessments.

The Suppressive Effects of Integrin Antibodies on the Infection of Hantaan Virus in Fibroblasts (한탄바이러스의 섬유아세포 감염에 대한 Integrin 항체의 억제 효과)

  • Park, Ho-Sun;Kim, Ki-Duk;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.55-66
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    • 1998
  • Pathophysiological mechanism of hemorrhagic fever with renal syndrome (HFRS) is not fully understood. Major clinical findings of HFRS patients are widespread hemorrhage, acute renal failure and shock. Basic lesion is vascular injury with microvascular hemorrhage and relatively little inflammation. According to autopsy findings, renal medulla shows focal hemorrhage, tubular necrosis and interstitial mononuclear infiltrates. The predominant cell type in the renal and pulmonary interstitium is a fibroblast and it participates in the healing process at the injury site by secreting a large amount of extracellular matrix proteins. Cultured human lung fibroblasts and Mongolian gerbil fibroblasts were known to be good host cells for the hantaan virus. It is possible that not only the endothelial cell but also the fibroblast is a target of Hantaan virus and the fibroblast might be involved in the pathogenesis and the healing process in HFRS. Integrins are adhesion molecules, and act as receptors for many extracellular matrix proteins. Recently, there are many reports that cell surface integrins influence on some viral infections or reversely viruses influence on the expression of integrins. The ${\alpha}_5{\beta}_1$ integrin is a major receptor for the fibronectin which is an important extracellular matrix protein secreted by fibroblasts. In this study, the role of ${\alpha}_5{\beta}_1$ integrin in the infection of Hantaan virus was examined by using anti-${\alpha}_5{\beta}_1$, integrin, anti-${\alpha}_5$ integrin and anti-${\beta}_1$, integrin antibodies in chicken embryo fibroblasts (CEF) and Mongolian gerbil fibroblasts(MGF). The treatment of anti-${\alpha}_5{\beta}_1$, integrin antibody in CEF reduced the virion titers 26.8% and the amount of nucleocapsid N protein 32.6% when compared with control CEF. When MGF were treated with anti-${\alpha}_5$, anti-${\beta}_1$ and anti-${\alpha}_5{\beta}_1$ integrin antibodies, virion titers were reduced by 26.5%, 29.4% and 28.7% and the amount of nucleocapsid N protein were reduced by 65.2%, 59.7% and 72.6%. These results suggested that ${\alpha}_5{\beta}_1$ integrin might act as a receptor for the Hantaan virus or blocking of ${\alpha}_5{\beta}_1$ integrin influences on the viral replication in CEF and MGF. It is also possible that the blocking of only one subunit of integrin represents similar results in that of whole molecule.

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Epidemiologic and clinical features in children with acute lower respiratory tract infection caused by human metapneumovirus in 2006-2007 (2006-2007년 소아 급성 하기도 감염증에서 유행한 메타뉴모바이러스의 유행 및 임상 양상)

  • Park, Gwi Ok;Kim, Ji Hyun;Lee, Jae Hee;Lee, Jung Ju;Yun, Sin Weon;Lim, In Seok;Lee, Dong Keun;Choi, Eung Sang;Yoo, Byoung Hoon;Lee, Mi Kyung;Chae, Soo Ahn
    • Clinical and Experimental Pediatrics
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    • v.52 no.3
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    • pp.330-338
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    • 2009
  • Purpose : The causes of acute lower respiratory tract infection (ALRTI) are mostly attributable to viral infection, including respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus A/B (IFV A/B), or adenovirus (ADV). Several Korean studies reported human metapneumovirus (hMPV) as a common pathogen of ALRTI. However, studies on seasonal distribution and clinical differences relative to other viruses are insufficient, prompting us to perform this study. Methods : From November 2006 to October 2007, we tested nasopharyngeal aspiration specimens in children hospitalized with ALRTI with the multiplex reverse transcriptase-polymerase chain reaction to identify 6 kinds of common pathogen (hMPV, RSV, PIV, IFV A/B, and ADV). We analyzed positive rates and clinical features by respiratory chart review. Results : We detected 38 (8.4%) hMPV-positive cases out of 193 (41.8%) virus-positive specimens among 462 patients. HMPV infection prevailed from March to June with incidence peaking in April. HMPV-positive patients were aged 15 years (76.3%), and the ratio of boys to girls was 1.2:1. The median age was 27 months. HMPV primarily caused pneumonia (76.3 %) (P=0.018). Average hospitalization of HMPV-associated ALRTI patients was 5.8 days. In addition, they showed parahilar peribronchial infiltration (100%) on chest X-ray, normal white blood cell count (73.7%), and negative C-reactive protein (86.8 %) (P>0.05). All hMPV-positive patients recovered without complication. Conclusion : HMPV is a common pathogen of ALRTI in Korean children, especially in 1-5 year olds, from March to May. Immunocompetent children diagnosed with hMPV-associated ALRTI may have a good prognosis.

Development of a Duplex RT-PCR Assay for the Simultaneous Detection and Discrimination of Avirulent and Virulent Newcastle Disease Virus (NDV) (뉴캣슬병 바이러스 검출 및 병원성 감별을 위한 Duplex RT-PCR법 개발)

  • Kim, Ji-Ye;Lee, Hyun-Jeong;Jang, Il;Lee, Hee-Soo;Yoon, Seung-Jun;Park, Ji-Sung;Seol, Jae-Goo;Kim, Seung-Han;Hong, Ji-Mu;Wang, Zillian;Liu, Hualei;Choi, Kang-Seuk
    • Korean Journal of Poultry Science
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    • v.44 no.2
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    • pp.93-102
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    • 2017
  • A duplex RT-PCR (dRT-PCR) assay was developed for the simultaneous detection and discrimination of non-virulent and virulent Newcastle disease virus (NDV) in a single PCR tube. Primers targeting the large polymerase protein (L) gene and the fusion protein (F) gene of NDV were designed to detect all NDVs (by common type PCR primers) and virulent NDVs (by pathotype PCR primers), respectively and evaluated experimentally with reference NDV strains and other poultry viral pathogens. PCR products of the expected size of 386 bp were amplified from all NDV samples whereas PCR products of the expected size of 229 bp were amplified from virulent NDV samples alone. Cross reaction was not observed with other avian viral pathogens. The detection limit of NDV by the dRT-PCR was estimated to be $10^3$ 50% egg infectious dose/0.1 mL. In the dRT-PCR using field isolates of NDV, the pathotype PCR primers detected specifically all of virulent field isolates of NDV from Malaysia, Pakistan and China whereas common type PCR primers detected 94.4% (51/54) of field isolates of NDV from China. Three Chinese NDV isolates with false negative result were non-virulent viruses. Our results indicate that the dRT-PCR might provide a rapid and simple tool for rapid simultaneous detection and discrimination of non-virulent and virulent NDVs. Therefore the developed dRT-PCR assay provides a powerful novel means for the rapid diagnosis of Newcastle disease.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Epidemiology of Acute Viral Lower Respiratory Tract Infection in Hospitalized Children in Two Different Areas of Korea (2개 지역에서 소아의 급성 바이러스성 하기도염의 유행성에 관한 역학조사)

  • Moon, Jeong Hee;Suh, Kyoung Jin;Chung, Eun Hee;Shin, Mee Yong;Lee, Ju Suk;Park, Yong Mean;Lee, Kwang Sin;Ahn, Kang Mo;Lee, Nam Yong;Ma, Sang Hyuk;Lee, Sang Il
    • Pediatric Infection and Vaccine
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    • v.9 no.2
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    • pp.193-200
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    • 2002
  • Purpose : This study was performed to investigate the epidemiology of viral acute lower respiratory tract infection(ALRI) in two different areas of Korea. Methods : A total of 796 patients hospitalized for ALRI aged 15 years or less from June 2000 to June 2001 in Samsung Seoul hospital(SSH) and Masan Fatima hospital(MFH) were enrolled. Viral etiologies were confirmed using nasopharyngeal aspirates. We compared etiologic agents, age distribution, clinical manifestations, and seasonal occurrence of viral ALRI between the two hospitals. Results : Virus was isolated in 208 patients(26.1%). The proportion of patients aged under 2 years in SSH was 60.2%, while those in MFH was 90.0%(P<0.05). Respiratory syncytial virus(RSV) was more prevalent in MFH, but adenovirus, influenza virus and parainfluenza virus were more prevalent in SSH(P<0.05). Croup and bronchiolitis occurred more frequently in MFH than in SSH(P<0.05). The most frequent viral pathogens causing bronchiolitis and croup were RSV and parainfluenza virus, respectively, in both hospitals. Adenovirus was the main cause of pneumonia in SSH, in contrast to RSV in MFH. In terms of tracheobronchitis, adenovirus was detected most frequently in SSH, whereas influenza virus-type A was mainly isolated in MFH. Similar pattern of seasonal occurrences of RSV, parainfluenza virus and influenza virus-type A was noted in both hospitals. Adenovirus was isolated sporadically throughout the study periods. Conclusion : Seasonal occurrence and clinical syndromes according to viral pathogens showed similar pattern in two areas. However, distribution of offending viruses was different, although this is mainly related to the different age distribution. An annual nationwide surveillance is necessary to understand the viral epidemiology associated with respiratory illnesses in Korea.

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Adenoviral Lower Respiratory Tract Infections in Children; Serotypes and Clinical Characteristics (소아의 아데노바이러스 하기도 감염증: 혈청형 및 임상상)

  • Shin, Seon Hee;Lee, Hoan Jong;Yoon, Hae Sun
    • Pediatric Infection and Vaccine
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    • v.9 no.2
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    • pp.182-192
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    • 2002
  • Purpose : This study was performed to characterize the epidemiologic and clinical features of acute adenoviral lower respiratory tract infections(LRTIs). Methods : Virological analysis was done from respiratory specimens obtained from patients with LRTIs hospitalized to other hospitals and referred to the Department of Pediatrics, Seoul National University Children's Hospital(SNUCH) from June 1998 to July 2000. Viral diagnosis was made by isolation of viruses employing HEp-2 cell culture and indirect immunofluorescent staining with monoclonal antibodies. Serotypes of adenoviruses were determined by neutralization test using antiserum for types 1, 2, 3, 4, 5, 6, 7 and 11. Medical records of children admitted to the SNUCH were reviewed retrospectively. Results : Adenovirus was isolated from 118(9.0%) of 1,305 children with LRTIs. Serotypes were 3(39.0%), 7(16.9%), 1(11.0%), 2(7.6%), 4(7.6%), 6(5.9%), 11(2.5%), and 5(0.8%) and 10 strains(8.5%) were not neutralized by antisera included in the study. Infections by type 3 and type 7 occurred in outbreaks. Male to female ratio was 1.0:0.9 and mean age was 1.95 years. The clinical diagnoses were pneumonia(83%), acute tracheobronchitis(12%) and bronchiolitis(5%). Associated symptoms, signs and abnormal laboratory findings included cough(100%), sputum(73.5%), fever(54.2%), rale(59.3%), wheezing(34%), anemia(35%) and leukopenia(15.8%). Mortality was in 13.5%. Residual radiologic sequelae was identified in 32.6% of the patients followed. Conclusion : These data confirms that adenovirus may cause severe lower respiratory tract diseases, and infections by type 3 and 7 may occured in outbreaks.

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Etiology and Clinical Manifestation of Acute Gastroenteritis in Children (소아 급성 위장관염에서의 원인과 임상양상)

  • Im, Ik-Jae;Lee, Mee Jeong;Chung, Eun Hee;Yu, Jeesuk;Chang, Young Pyo;Park, Woo Sung;Park, Kwisung;Song, Nak Soo;Baek, Kyung Ah;Cha, Yune Tae
    • Pediatric Infection and Vaccine
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    • v.13 no.2
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    • pp.147-155
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    • 2006
  • Purpose : The purpose of this study is to evaluate epidemiological data of pathogens obtained from stool exams and compare them with the clinical course in pediatric patients with symptoms of acute gastroenteritis. Methods : Subjects were selected from patients presenting with symptoms of acute gastroenteritis who visited the outpatient clinic or who were admitted to the Dankook University Hospital from December of 2004 to December of 2005. Stool exams for 17 pathogens was performed. RT-PCR was used to detect norovirus and enzyme-linked immunoabsorbant assay (ELISA) was used to detect rotavirus, adenovirus and astrovirus in the subjects stool samples. Ten different species of bacteria(Salmonella spp., Shigella spp., Clostridium perfrigens, Campylobacter spp., Escherichia coli, Vibrio spp., Staphylococcus aureus, Bacillus cereus, Yersinia spp., and L. monocytogenes) were each selectively cultivated and enzyme immunoassays(EIA) was used to test for antigens for C. parvum, E. histolytica and G. lamblia. Retrospective chart review was performed for comparisons of clinical manifestations. Results : A total of 215 subjects was selected and of these 89 cases(41.4%) showed positive results for at least one pathogen. Male to female ratio was 1.3:1. Age distribution showed 4 cases less than one month(4.5%), 4 cases from 1~2 months(4.5%), 24 cases from 3~12 months(26.7%), 47 cases form 13~48 months(52.8%), 10 cases greater than 48 months (21.2%). Viruses showed the greatest proportion of cases with 68 subjects(77.5%), of these rotavirus being the most commonly reported in 50 cases. Bacteria was identified in 26 cases (29.2%), of these nontyphoidal salmonella was noted in 10 cases. Protozoa followed with 21 cases(23.6%), of these C. parvum was noted in 11 cases and G. lamblia was noted in 10 cases. Mixed infections with more than two pathogens were seen in 22 cases(24.7%), of these viral infection with accompanying parasitic infection was seen in 12(54.5%) cases. Conclusion : In this study we examined various pathogens known to cause acute gastroenteritis in children. Further studies for various pathogens can provide useful information for management of the acute gastroenteritis.

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Cross-Protection Effectiveness of Cucumber mosaic virus (CMV) Isolates Associated with Satellite RNA for Prevention of CMV Disease in Pepper Plants (Satellite RNA 보유 Cucumber mosaic virus(CMV)의 고추 CMV병에 대한 교차방어 효과)

  • 최장경;성미영;정혜진;홍진성;이상용
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.155-163
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    • 2001
  • Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.

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Construction of a Fluorescently Labeled Infectious R Peptide-Less Moloney MLV Molecular Clone for Analysis of Syncytium (합포체 분석을 위해 R 펩타이드가 결여된 형광 표지 Moloney 마우스레트로바이러스 Molecular Clone 제조)

  • Lee, Yong-Jin;Park, Jin-Woo;Lee, Kyu-Jun;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.246-250
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    • 2009
  • Retroviruses enter host cells by membrane fusion between the viral Env proteins on the virus membrane and a virus receptor on the cellular membrane. The envelope protein of the ecotropic Moloney murine leukemia virus is synthesized as a gp85 precursor and is proteolytically cleaved into an extracellular surface unit (SU) and the transmembrane protein (TM). The cytoplasmic tail (16 amino acid; R peptide) of the TM protein is further cleaved by the viral protease during virion maturation. Unlike the wild type Env protrin bearing the R peptide, R peptide-truncated Envelope induces syncytia in susceptible cells. To understand the mechanism of R peptidetruncated Env in syncytium formation, R peptide-truncated Env expressing full-length molecular clone containing EGFP in PRR (proline rich region) of Env was constructed. This molecular clone induced syncytia in transfected NIH3T3 cells, fluorescence was detected in the cytoplasm and at the plasma membrane, while the nuclei did not stain and appeared black by fluorescence microscopy. Interestingly, virions with truncated envelope produced from transfected NIH3T3 cells induced syncytia in NIH3T3 cells, but fluorescence was not detected in the same infected cells. It is believed that cell-free viruses direct the fusion of neighboring cells without infection. Our data suggests that use of EGFP-tagged envelope for monitoring syncytium is a sensitive and convenient method. We also found that virion incorporated the R peptide-truncated Env is able to induce the formation of syncytia by fusion from without.