• Journal of Microbiology and Biotechnology
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• v.24 no.8
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• pp.1044-1050
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• 2014

Since there is no consensus about the most reliable assays to detect invasive aspergillosis from samples obtained by minimally invasive or noninvasive methods, we compared the efficacy of an enzyme-linked immunosorbent assay (ELISA) for galactomannan (GM) detection and quantitative real-time PCR assay (qRT-PCR) for the diagnosis of invasive pulmonary aspergillosis. Neutropenic, male Sprague-Dawley rats (specific pathogen free; 8 weeks old; weight, $200{\pm}20g$) were immunosuppressed with cyclophosphamide and infected with Aspergillus fumigatus intratracheally. Tissue and whole blood samples were harvested on days 1, 3, 5, and 7 post-infection and examined with GM ELISA and qRT-PCR. The A. fumigatus DNA detection sequence was detected in the following number of samples from 12 immunosuppressed, infected rats examined on the scheduled days: day 1 (0/12), day 3 (0/12), day 5 (6/12), and day 7 (8/12) post-infection. The sensitivity and specificity of the qRT-PCR assay was 29.2% and 100%, respectively. Receiver operating characteristic curve (ROC) analysis indicated a Ct (cycle threshold) cut-off value of 15.35, and the area under the curve (AUC) was 0.627. The GM assay detected antigen in sera obtained on day 1 (5/12), day 3 (9/12), day 5 (12/12), and day 7 (12/12) post-infection, and thus had a sensitivity of 79.2% and a specificity of 100%. The ROC of the GM assay indicated that the optimal Ct cut-off value was 1.40 (AUC, 0.919). The GM assay was more sensitive than the qRT-PCR assay in diagnosing invasive pulmonary aspergillosis in rats.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.1041-1050
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• 2009

Equine respiratory disease is a common cause of poor performance and training interruptions. The higher incidence rate of infectious upper respiratory disease (IURD) in thoroughbred racehorses at the Seoul Race Park coincided with the frequent stabling season, shorter stabling periods, and younger ages in this study. Incidence rates were also correlated with significantly lower proportions of cells expressing MHC class II-, CD2 antigen-, $CD4^+$- or $CD8^+$-T lymphocyte-, and B lymphocyte in IURD patients compared with healthy control groups in the summer and fall and in 2-and-3-year-old groups. The data suggested that movement and new environments may have resulted in immunosuppression and inappropriate responses to respiratory pathogens in IURD patients. The IURD incidence decreased with age, perhaps by the acquisition of immunity, and study results suggested that immunologic protection was associated with IURD, particularly in young thoroughbred racehorses. Streptococci isolates were identified in 11 of 72 IURD horses, and 3 of these isolates were identified as Streptococcus. equi subsp. equi. S. equi subsp. zooepidemicus was isolated from 2 of 23 IURD horses in the spring (8.7%), 5 of 23 in the summer (21.7%), and 1 of 6 in winter (16.7%). S. equi subsp. zooepidemicus (5%) was also identified in 3 of 61 isolates from clinically normal horses. Racetracks should implement anti-IURD protective measures by assessing the capacity of equine immunologic protection at the Park and by limiting the introduction of specific respiratory pathogens (such as S. equi subsp. equi) by preventing the access of infected but subclinical horses with a specified respiratory pathogen-free certification system prior to Park entry.

• Korean Journal of Poultry Science
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• v.37 no.2
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• pp.159-165
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• 2010

Field strains of Ornithobacterium rhinotracheale (OR) were tested on their virulence in specific pathogen free (SPF) embryonated chicken eggs and 3-week-old broilers. When infected with three different OR isolates (OR-161, OR-240 and OR-295) through yolk sac infection route, all strains appeared to be highly pathogenic with responsible mortality 66% and 100% within 12 days post infection (DPI). To test the virulence of OR in the commercial broilers, 3 week-old broilers were grouped depends on the inoculation route of OR isolate (OR-295) through five different infection routes; group 1 (IT: intratracheal), group 2 (IM: intramuscular), group 3 (IV: intravenous), group 4 (aerosol) and group 5 [Mixed: NDV (LaSota)+OR aerosol]. Within 5 to 7 days after inoculation, only broilers given NDV+OR were slightly depressed and coughing, and had mild facial redness. Grossly, foamy and yellow-white yogurt like exudate in the air sacs, predominantly in the abdominal air sacs was present. In histology, infiltration of the air sac epithelium and lamina propria by macrophage and polymorphonuclear granulocytes was seen with cell debris and inflammatory cells, correlated with the presence of OR antigen, as demonstrated by immunohistochemistry. Field strains of OR were able to induce high mortality in the embryonated chicken eggs, whereas broilers were less susceptible to OR infection. Interestingly, in the absence of NDV infection, the four groups of OR single infection only different route showed minimal and temporary microscopic air sac lesions. Thus, Newcastle disease virus (LaSota strain) showed triggering effects on the OR infection in chickens.

• Korean Journal of Veterinary Research
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• v.42 no.2
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• pp.241-251
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• 2002

Modified-live (ML) infectious laryngotracheitis (ILT) vaccines have been widely used as a preventive measure in Korea since the first outbreak of ITL. Recently, it has been observed that chickens vaccinated with the commercially available ML ILT vaccine have sometimes exhibited adverse clinical signs. In this study, we evaluated the quality of the vaccines by comparing titer of each vaccine batch and testing the stability of ILT virus (ILTV) in vaccine diluents and compared the safety and efficacy of vaccines in specific pathogen free (SPF) chickens. The ratio of maximum titer to minimum titer of vaccine produced by most manufacturers was 2 to 15. However, 2 out of 11 manufacturers produced vaccines of which the ratio was 74 to 478. Most vaccines examined were maintained vaccine titers suitable for national regulations within expiry date. However, some vaccines did not keep the titer required for the national regulations. In the test for stability of ILTV in various diluents, ILTV was highly stable in lactose-phosphate-glutamine-gelatin solution, sucrose-phophate-glutamine-albumin solution and some vaccine diluents produced by manufacturers. The safety of ML ILT vaccines was assessed in 10-day-old SPF chicks. Mortality in SPF chicks inoculated intratracheally with one dose of vaccine varied depending on vaccines and some vaccines produced 50-85% mortality. Seven-week-old SPF chickens were vaccinated intraocularly with ML ILT vaccines and then challenged intratracheally with ILT challenge virus 14 days after vaccination. The protection rate was assessed by clinical signs and reisolation of the ILT challenge virus from tracheas taken at day 4 after challenge. There were slight respiratory reactions in some vaccinated chickens after vaccination but these reactions disappeared within 5 days after vaccination. No further clinical signs and death were observed. Protection rate determined by clinical signs and mortality was 100% in all vaccinated groups. However, the challenge virus was isolated from all tracheas of chickens vaccinated with vaccine B or control groups. The challenge virus was also isolated in the trachea of one in five chickens vaccinated with either vaccine F or K, but not in tracheas of chickens vaccinated with other vaccines. In the present study, the stability of vaccine diluents, pathogenicity and protection rate based on reisolation test of the challenge virus were different depending on vaccines produced by eleven manufacturers.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.359-374
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• 2000

Cyclosporine A(CsA) is a widely used immunosuppressant for transplant patients and is also used for the treatment of a wide variety of systemic diseases with immunologic disorders. However, its use is frequently limited because of complications such as nephrotoxicity or gingival hyperplasia. Although several hypotheses have been postulated for CsA-induced gingival hyperplasia, i.e. various cytokine effects of inflammatory cells, existence of plaque or CsA itself, but its pathogenesis is still unclear. For experimental chronic CsA toxicity, salt depletion has been shown to increased susceptibility of rodents to the effects of CsA, and this maneuver facilitates production of arteriolopathy and interstitial fibrosis in kidney that mimic the changes found in human. The purpose of this study was to evaluate pathogenesis of CsA-induced gingival hyperplasia by comparing changes between CsA administration groups of normal standard diet and those of low salt diet group. Specific pathogen-free, 20 to 25 days old(120 to 150 g), male Fisher-344 rats(KIST, Korea), 120 to 150g of body weight, were assigned to four groups of six animals each after one week of adaptation period for powder food. Group 1 received olive oil($300{\mu}l/g\;of\;diet$) with normal standard diet(0.4% of sodium)(NSD). Group 2 received CsA(Cypol-N, Jonggundang, Korea; $300{\mu}g/g\;of\;diet$) with normal standard diet(NSD+CsA). Group 3 received same amount of olive oil with low salt diet(0.05 % of sodium, Teklad Premier, U.S.A.)(LSD). Group 4 received same dose of CsA with low salt diet(LSD+CsA). Rats were pair fed and were sacrificed after six weeks. Renal histologic lesions associated with CsA, consisted of cortical interstitial fibrosis, tubular atrophy and hyalinization of arterioles and the impairment of renal function including increase of serum creatinine and decrease of glomerular filtration rate was more severe in low salt diet group. These were proved as the results of activated of renin-angiotensin system in the kidney by low salt condition. Meanwhile the degree of gingival hyperplasia at incisor and molar tooth was less severe in low salt diet group compared with normal sodium diet group. Hyperplastic gingiva showed mild epithelial hyperplasia and expanded underlyng stroma which consisted of matrix increasement, capillary proliferation and dilatation. While the number and the activation of fibroblasts were increased, inflammatory cells were rare in the stroma. The immunohistochemistry for TGF-${\beta}_1$ in the kidney and gingiva revealed stronger positive in LSD+CsA in kidney but in gingiva of NSD+CsA. These results suggested followings; Gingival hyperplasia can be developed without inflammatory cells infiltration and seemed not induced by CsA by itself. The major role for gingival hyperplasia by CsA would be the secondary effect of TGF-${\beta}$, which maybe upregulated by CsA administration. Low salt diet can attenuate this hyperplasia perhaps by decreasing the activation of $TGF-{\beta}$.

• Journal of Oriental Neuropsychiatry
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• v.20 no.2
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• pp.111-120
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• 2009

Objectives : This study was performed to investigate the effects of Needle Electrode Electrical Stimulation (NEES) on ischemia-induced cerebrovascular accidents. After obstruction and reperfusion of ${\ast}{\ast}$ arteries in white mice, the amounts of necrosis and inflammation related substances IL-6, Caspase-3, and PARP, C-fos were measured in neurons of the hippocampus. The following results were obtained. Methods : This study used 21 male specific pathogen free (SPF) SD (Sprague Dawley) rats, 8 weeks of age and approximately 300 g in weight, that were given at least 1 week to adapt to the lab environment Each exposed artery was completely occluded with non-absorbent suture thread and kept in that state for 5 minutes. The sutures were then removed to allow reperfusion of blood. Test group is control group for comparison with the common carotid artery occlusion models, a GI group that underwent common carotid artery occlusion, and a needle electrode electrical stimulation (NEES) group that underwent NEES after artery occlusion. The GI and NEES groups were given 12, 24, or 48 hours of reperfusion before NEES. NEES device (PG6, ITO, Japan, 9V, current, 2Hz) was used to stimulate the right and left acupoint ST36 of the SD rats for 30 minutes while they were sedated with 3% isoflurane. An immunohistochemistry test was done on the forebrains of the GI induced rats. All the data collected from this study was symbolized and analyzed using a statistics processing program (SPSS 12.0K/PC). The level of significance was set at ${\alpha}$=0.05 and a T-TEST analysis was used to find out the effects of treatment on each of the groups: the normal group, the CVA induced group, and the treatment after CVA induction group. Results : Both PARP and C-fos immuno-reactive cells, related to apoptosis, were greater in the GI groups than the NEES group. Caspase and IL-6 immuno-reactive cells, related to inflammation, were greater in the GI and NEES groups than the control group. Conclusions : This research was conducted to study the effects of NEES on CVA due to ischemia. Occlusion and reperfusion was performed on the common carotid arteries of white rats, after which amounts of substances related to neuron necrosis and inflammation - PARP, IL-6, Caspase-3, and C-fos - were measured in the Hippocampus

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.6
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• pp.1034-1041
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• 2010

The oriental medicine Jangwonhwan, which is a boiled extract of 12 medicinal herbs/mushroom, has been prescribed for patients with cognitive dysfunction and it is originally from the Korean medical text, DonguiBogam(amnesia chapter). Recently, a modified recipe of Jangwonhwan (LMK02-Jangwonhwan) consisting of seven medicinal plants/mushroom, was shown to reduce ${\beta}$-amyloid deposition in the brain of Tg-APPswe/PS1dE9 mouse model of Alzheimer disease. The toxicity of LMK02 was investigated in SD rats by oral repeated adminstration for 4 weeks and we tried to determine test does for 13 weeks repeated study. Quality control of tablet form of LMK02 was established by estimating indicative components, Ginsenoside Rg3 of Red Ginseng and Decursin of Angelicagigas Nakai. The toxicity of LMK02 was investigated in 6 weeks old specific pathogen free (SPF) Sprageu-Dawley rats by oral administration. Each test group were consist of 5 male and 5 female and they received doses of 500, 1,000 and 2,000 mg/kg/day of test substance for 4 weeks. The clinical signs, death rate, body weight, food consumption, ophthalmic examination, urinalysis, hematological and serum biochemistry, organ weight and pathological changes were examined and compared with those of control group. Urinalysis : We observed increase of PRO(p<0.01), SG(p<0.01) in female rats of 1,000 mg/kg/day and 2,000 mg/kg/day(p<0.01). Also, we observed increase of pH and KET in female rats of 1,000 mg/kg/day(p<0.05) and of 2,000 mg/kg/day(p<0.01). WBC in female rats in 1,000 mg/kg/day and 2,000 mg/kg/day were on increase. Hematological test : We observed increase of MCV in male rats of 250 mg/kg/day. (p<0.05) Serum biochemistry test : We found increase of CHO in female rats of 2,000 mg/kg/day(p<0.05). During the experimental period, there were no animals dead or moribund. There were no treatment related changes of general symptom, food and water consumption, organ weight and autopsy According to the results of 4-week repeated dose range finding study, the highest dose was established as 1000 mg/kg for 13-week repeated dose toxicity study and we determined to put 2 more groups by common ratio two.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.117-129
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• 2000

This study was carried out to examine proliferative activity and expression of c-myc oncoprotein and p2lras in normal and preneoplastic rat livers induced by an in vivo mid-term chemical carcinogenesis assay. Sixty, six-week-old male specific pathogen free Sprague-Dawley male rats were randomly divided into five groups. Group I was received a single intraperitoneal(IP) dose(200mg/kg) of diethylnitrosamine(DEN). Group 2(10 rats) was operated partial hepatectomy(PH) and Group 3 was received IP(200mg/kg) DEN, fed two weeks later with 500ppm of phenobarbital(PB). Group 4 was received IP(200mg/kg) DEN, fed two weeks later 500ppm(PB) and PH at week 3 after the onset of experiment. While group 5(20 rats) was not treated and used as a control group. All the rats were sacrificed at age 14 weeks except 10 rats from group 5 were sacrificed at the onset of experiment. Livers of all rats were examined for 5-bromo-2'-deoxyuridine(BrdU) incoporation, proliferating cell nuclear antigen(PCNA), silver-binding nucleolar organizer regions(AgNORs) counts per nucleus and expression of c-myc oncoprotein and p21ras. Both the number and area of the preneoplastic lesions were significantly(p<0.01) compared to other groups. A significant(p<0.01) increase in immunoreactive cells were detected in preneoplastic hepatocytes in Groups 3 and 4 by PCNA and BrdU immunohistochemical stain. The number of the positive cells were significantly(p<0.05) lower in normal 14-week-old rats than those of 6-week-old rats. The results showed that proliferative activity of the hepatocytes was increased by treatment with DEN, PH and PB. Meanwhile, AgNORs counts per nucleus were significantly(p<0.05) increased in the preneoplastic hepatocytes of rats in both groups 3 and 4. The expression of c-myc oncoprotein and p21ras were more readily localized within the hepatic preneoplastic lesions such as hyperplastic nodules. Especially, group 4 showed significantly (p<0.05) overexpressed levels compared to groups 1 and 3. These findings suggest that PCNA, BrdU and AgNORs are significantly increased and c-myc oncoprotein and p21ras are significantly overexpressed in hepatic preneoplastic lesions induced by mid-term carcinogenesis. So these parameters can be an effective markers for hepatic prencoplastic lesions.

• Journal of Applied Biological Chemistry
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• v.59 no.4
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• pp.351-356
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• 2016

Bacterial blotch caused by Pseudomonas tolaasii is one of the major diseases of oyster mushroom, Pleurotus ostreatus. Application of bacteriophages is a very useful tool to decrease the density of pathogens and it has been successful to making disease-free cultivation area, known as phage therapy. Effect of phages on pathogen sterilization is very limited to the specific host strains. Minor variations of the host strains may cause changes in phage sensitivity. The phage-resistant strains of P. tolaasii were isolated and their pathogenic characters were investigated to improve the effectiveness of phage therapy. In the phylogenetic analysis, both phage-resistant strains and the corresponding host strains were identical based on the sequence comparison of 16S rRNA genes. The pathogenic characters, such as hemolytic activity and brown blotch formation, were measured on the phage-resistant strains and no correlation between phage-resistance and pathogenic characters was observed. Nevertheless, pathogenic characters were sometimes changed in the phage-resistant strains depending on the host strains. In order to make the phage therapy successful, the bacteriophages having a wide host range should be isolated.

• Korean Journal of Agricultural Science
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• v.38 no.4
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• pp.681-687
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• 2011

Bioorgan transgenic cloned mini pig has a problem of growth retardation in uterus during end of pregnancy so that survival rate is very low in newborn piglet. In order to support their life after birth, cesarean section of fetus with sufficient growth in uterus was tested in this study. First of all, fetus growth measured using a ultrasound scanner during pregnancy in transgenic mini pig, comparing normal pig. After 113 days for delivering, fetus was removed out of uterus. Fetus growth for normal pig was 1.8 cm at 4weeks and 14.4 cm at end of pregnancy (15 weeks). At 113 days, fetus growth was $15.9{\pm}4$ cm in ultrasound scanner and real growth measurement from fetus removal out of uterus was $16.0{\pm}2$ cm. It is very a similar result between measurement of ultrasound scanner and real measurement. Therefore, using ultrasound scanner for measuring fetus growth will be useful to predict fetus growth in uterus.