Barley yellow mosaic virus (BaYMV) is transmitted by a root-inhabiting Polymyxa graminis (P. graminis) and thus the disease is called "soil-borne". In this study, the presence of P. graminis was confirmed by PCR test based on specific sequences of P. graminis type I (P. graminis f. sp. temperata) Internal Transcribed Spacer (ITS). P. graminis was detected in the BaYMV infected soil and root of barley plants. The monitoring of P. graminis was conducted in March 2015 in 8 korean provinces. It was detected in the soil of all collected regions. This is the first report on a P. graminis a survey of Korea.
This bio-acoustic study was aimed at classifying the different porcine wasting diseases through sound analysis with emphasis given to differences in the acoustic footprints of coughs in porcine circo virus type 2 (PCV2), porcine reproductive and respiratory syndrome (PRRS) virus and Mycoplasma hyopneumoniae (MH) - infected pigs from a normal cough. A total of 36 pigs (Yorkshire${\times}$Landrace${\times}$Duroc) with average weight ranging between 25-30 kg were studied, and blood samples of the suspected infected pigs were collected and subjected to serological analysis to determine PCV2, PRRS and MH. Sounds emitted by coughing pigs were recorded individually for 30 minutes depending on cough attacks by a digital camcorder placed within a meter distance from the animal. Recorded signals were digitalized in a PC using the Cool Edit Program, classified through labeling method, and analyzed by one-way analysis of variance and discriminant analysis. Input features after classification showed that normal cough had the highest pitch level compared to other infectious diseases (p<0.002) but not statistically different from PRRS and MH. PCV2 differed statistically (p<0.002) from the normal cough and PRRS but not from MH. MH had the highest intensity and all coughs differed statistically from each other (p<0.0001). PCV2 was statistically different from others (p<0.0001) in formants 1, 2, 3 and 4. There was no statistical difference in duration between different porcine diseases and the normal cough (p>0.6863). Mechanisms of cough sound creation in the airway could be used to explain these observed acoustic differences and these findings indicated that the existence of acoustically different cough patterns depend on causes or the animals' respiratory system conditions. Conclusively, differences in the status of lungs results in different cough sounds. Finally, this study could be useful in supporting an early detection method based on the on-line cough counter algorithm for the initial diagnosis of sick animals in breeding farms.
Kim, Jae-Hyun;Kuem, Wan-Soo;Lee, Sin-Ho;Kim, Jeong-Soo;Jeon, Yong-Ho;Jung, Suk-Hun;Chung, Youl-Young;Park, Yong-Hack
Journal of the Korean Society of Tobacco Science
/
v.28
no.2
/
pp.100-110
/
2006
A mutant of Potato vims Y (PVY) was occurred in PVY resistance flue-cured tobacco breeding line KF0402 $(TC1146{\times}KF117)$ showing vein necrosis at Suwon in Korea. This isolate, PVY-SWM, was differentiated from other PVY based on biological properties and nucleotide sequence analyses of coat protein gene. PVY-SWM caused typical symptoms on 21 indicator plants as compared to the PVY-TOJC37. Remarkably, the PVY-SWM induced distinctly different symptom of systemic vein necrosis on tobacco cultivars V.SCR, PBD6, TN86, TN90, Virgin A Mutant (VAM), Wislica, NC744, KB108 and KB111, which were reported to have the recessive potyvirus resistance gene va. In RT-PCR assays with specific primers for detection of PVY, a single band of about 800bp in length was produced. The amplified DNA was cloned and the nucleotide sequence was determined. The coat protein gene of PVY-SWM showed 88.4%-99.0% and 92.5%-98.5% identities to the 12 different PVY isolates of Genbank Database at the nucleotide and amino acidi respectively. Multiple alignments as well as cluster dendrograms of PVY-SWM isolate revealed close phylogenetic relationship to the $PVY^{NTN}$ subgroup.
The Journal of Korean Association of Computer Education
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v.13
no.6
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pp.91-98
/
2010
NAC(Network Access Control) has been developed as a solution for the security of end-point user, to be a target computer of worm attack which does not use security patch of OS and install Anti-Virus, which spreads the viruses in the Intra-net. Currently the NAC products in market have a sufficient technology of pre-connect, but insufficient one of post-connect which detects the threats after the connect through regular authentication. Therefore NAC users have been suffered from Zero-day attacks and malware infection. In this paper, to solve the problems in the post-connect step we generate the normal behavior profiles using the traffic information of each host, host information through agent, information of open port and network configuration modification through network scanner addition to authentication of host and inspection of policy violation used before. Based on these we propose the system to detect the hosts infected malware.
In this paper, a in situ hybridization(ISH) has been used to investigate the yield of viral RNA expression from each organ tissues. It is studied to establish a rapidly, specific diagnostic method detecting rabbit haemorrhagic disease virus(RHDV) RNA in 10% formalin-fixed, paraffin-em-bedded tissues of naturally RHDV-infected rabbits using oligonucleotide probe to be made by RHDV total sequences. Biotin was used as the oligonucleotide probe marker. in situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method. All ISH procedure of RHDV were completed to Mi-croProbe$^{TM}$ capillary action system within 1-2 hours. In this report, RHDV was distributed widely in the cytoplasm of liver cell and the cortex of kidney but lung tissue and medulla of kidney were showed to positive reaction at locally. Although not entirely free of technical limitations, nucleic acid identification holds advantages over other diagnostic tests, including exquisite sensitivity, specificity, interchangeability and speed. It is expected that, in the immediate future viral nucleic acid detection will be a prominent part of the methods used in histopathology.
Porcine cytomegalovirus (PCMV) is a betaherpesvirus which causes reproductive failure in breeding sows and generalized infection in newborn piglets. It has worldwide distribution including Korea. Serological survey on this virus has been reported in 76.3% of pigs, but virological survey and epidemiological analysis on PCMV distribution have been reported in only a few papers in Korea. In this study, we investigated the virological prevalence and infection status of PCMV on a farm level in selected swine farms with respiratory diseases. A total of 1,938 blood samples taken from groups of pigs of different ages were collected from 31 farms distributed nationwide in 2006 and 2007 and tested by PCR to detect the presence of PCMV. Virological prevalence at farm level and pig level were 96.8% and 17.5%, respectively, suggesting that PCMV has endemically infected Korean pig herds. The prevalence at farm level in gilts, sows and suckling piglet groups were 16.7%, 36.7% and 56.7%, indicating that vertical infections frequently occurred in conception or newborn stage. Thereafter, detection rates of PCMV were slightly increased in pig groups aged 40 and 70 days (70.0% and 73.3%), and then gradually decreased as they aged - 33.3% in 100, 26.7% in 130 and 16.7% in 160 day old pig groups. The prevalence at pig level has similar patterns to that at farm level. With the passage of time, the variation of infection patterns of PCMV was investigated in four PCMV-positive farms. Three blood samples were collected at intervals of 6 months in each farm, and examined for presence of PCMV using PCR. The results revealed that once PCMV was introduced to the pig farms, it continuously circulated between and within groups of sows and piglets in those farms. Taken together, it can be concluded that PCMV has endemically infected Korean pig farms and has the potential risk for emerging pathogen in combination with the known endemic pathogens including porcine reproductive, respiratory syndrome virus and porcine circovirus type 2. Therefore, more research is needed on diagnosis, epidemiology and control strategy for PCMV on the field.
This study was conducted to assess the predictive value of p-glycoprotein (p-gp) and p53 immunoexpression in human papillomavirus (HPV) infected cases of cervical dysplasia. Expression of both p-gp and p53 proteins was detected in cervical smears from 177 squamous intraepithelial lesions (SIL) cases along with 183 "atypical squamous cells of unknown significance" (ASCUS) and 150 normal cases. HPV 16 and 18 infection was detected by polymerase chain reaction using type-specific primers for HPV sub-types. There were no significant detectable p53 and p-gp expression in the normal cervix smears (p>0.05). In the ASCUS group 10 cases were positive for both p53 and p-gp immunoreactivity. In cervical dysplasia cases, p53 was positive in 86 (48.58%) while p-gp was positive in 93 (52.54%) and the two markers showed a highly significant correlation (r=0.92, p<0.001). Expression of p53 and p-gp was associated with grade of SIL (p<0.001). A positive correlation between the presence of HPV and expression of proteins p53 and p-gp in smears of patients with cervical lesions was also noted (p<0.001). Thus, p53 and p-gp immunostaining in cervical smears may act as an auxiliary biomarker for detection of HPV-associated cervical lesions. Additionally, a significant positive correlation between ascending grades of SIL and labeling indices of markers suggests that p53 and p-gp can be used as an adjunct to cytomorphological interpretation of conventional cervical Pap smears.
Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.
Three cDNA fragments located within NS5 region of HCV were synthesized by RT using viral RNA extracted from blood sample of Korean patient as a template. The cDNAs were amplified by PCR, cloned into the T-vector, and the nucleotide sequences were determined. Comparative analysis of the nucleotide and amino acid sequence of NS5 cDNAs showed that it is closely related with HCV type 1b. The cloned NS5 cDNA showed 91-94% homology at the nucleotide sequence level and 96-98% homology at the amino acid sequence level with several strains of the HCV type 1b. The NS5 cDNAs were subcloned into E. coli expression vectors to construct pRSETA5-1, pTHAN5-1, pRSETC5-2, pRSETBB1, pRESTCB1 and pRSETB-H3. Expression of the NS5 proteins was achieved by inducing the promoter with isopropyl-thio-${\beta}$-D-galactoside (IPTG) and confirmed by SDS-polyacrylamide gel electrophoresis. The NS5 proteins were immunoreactive against sera from Korean hepatitis C patients in Western blot analysis. Among the recombinant NS5 proteins, pRSETAS-1 plasmid derived protein, coded from aa2022 to aa2521 of HCV polyprotein, showed the strongest immunoreactivity against sera from Korean hepatitis C patients in immunoblot analysis. These results suggest that NS5 proteins would be useful as an antigen for detection of antibody against HCV in the blood samples.
Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.
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