• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.80-80
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• 1993

(목적) 본 연구에서 사용된 바이러스는 HIV-1 nef유전자가 일부 삭제되고 대신 Chloramphenicol acetyltransferase(CAT)가 pSVCAT recombinant 바이러스다. 이러한 recombinant 바이러스를 사용하는 이유는 첫째, CAT activity가 매우 민감하므로 바이러스의 복제억제 정도를 정확하게 측정 할 수 있고 둘째, simian immunodeficiency virus(SIV)의 경우 nef 유전가 in vivo에서는 바이러스의 복제에 필수적이므로 HIV가 SIV와 유사한 것으로 미루어 본 연구에서 사용되는 recombinant SVCAT 바이러스가 안전한 것으로 고려되기 때문이다. (방법) 특히 화합물이 HIV-1의 복제에 얼마나 영향이 있는가는 1) 어느정 도의 virus inoculm을 넣었는지 2) 사용하는 cell line 3) 사용한 cell line의 infection kinetics 4) 실험의 지속기간 5) 테스트하는 assay의 sensitivity에 의존한다. 따라서 $10^{5}$ cell의 H9과 sup T1을 24 well plate에 넣고 sup T1 cell line의 경우 3일 후 항 화합물에 의한 syncytia 형성 및 CAT activity의 억제정도를 현재 AIDS drug으로 쓰이고 있는 Zidovudine을 control로 비교 관찰하였다. H9 cell line의 경우 3일 간격으로 media의 3/4을 fresh media로 바꾸어 주고 9일 후 CAT assay를 하였다. 이러한 assay에서 activity를 보이는 화합물을 reverse transcriptase와 P24 ELISA assay를 재확인하였다.다.

• Journal of Veterinary Clinics
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• v.40 no.4
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• pp.298-302
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• 2023

A 7-year-old neutered male, domestic shorthair cat presented anorexia and lethargy. The complete blood cell count revealed severe non-regenerative anemia, lymphocytic leukocytosis, neutropenia, and thrombocytopenia. On the peripheral blood smear examination, medium to large lymphoblastic cells with moderate amounts of basophilic cytoplasm were observed in up to 70% of peripheral leukocytes. Feline leukemia and immunodeficiency viruses were not detected using a commercial diagnostic kit. While splenomegaly and blunt margins of the caudoventral liver were observed in abdominal radiography, changes in the intra-abdominal lymph nodes were not remarkable. Ultimately, flow cytometric immunophenotyping from the peripheral blood revealed a negative for B-cell markers (CD21-/CD79a-) and T-cell markers (CD3-/CD4-/CD5-/CD8-). Based on the hematological examination and the immunophenotyping assay, the cat was diagnosed with non-B, non-T acute lymphoblastic leukemia. Here, we report a rare case of non-B, non-T acute lymphoblastic leukemia to raise awareness and provide information on clinical symptoms and laboratory test and immunophenotyping analysis results.

• Journal of Veterinary Clinics
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• v.35 no.6
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• pp.273-275
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• 2018

A 6-month-old male cat was presented for investigation of depression, loss of appetite, dehydration, pale conjunctival mucous membrane, weight loss, fast heart and respiratory rates, nasal discharge and cough. Nasal swabs collected from the studied cat. As the results of bacterial culture with nasal swabs, it was suspected with Mycoplasma spp. Also, Mycoplasma species was detected by the PCR reaction with Mycoplasma genus primers. At species PCR assay, the specimens evaluated for the presence of M. felis, M. arginini, M. gateae, and Acholeplasma laidlawii and the result was visualization of bands from 238 bp in agarose gel 1.5% showing M. felis amplicons in samples. In conclusion, we detected M. felis in a cat with respiratory disease. PCR was able to detect successfully M. felis infection in cats.

• Journal of Korean Ophthalmic Optics Society
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• v.20 no.1
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• pp.75-81
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• 2015

Purpose: The present study was conducted to investigate whether the changes in structure and activity of antioxidative enzymes, superoxide dismutase(SOD) and catalase(CAT) present in the eyes appeared when they were repeatedly exposed to UV-A, and reveal the correlation of these changes. Methods: Each enzyme solution was prepared from the standardized SOD and CAT, and repeatedly exposed to UV-A of 365 min under the condition of 30 minutes, 1 hour and 2 hours a day over 1, 2, 3, 4 and 5 days. Structural denaturation of SOD and CAT induced by repeat UV-A irradiation was confirmed by the electrophoretic analysis, and their enzyme activity was determined by the colorimetric assay using the proper assay kit. Results: SOD exposed repeatedly to UV-A showed the polymerization pattern through the electrophoretic analysis when it was repeatedly exposed under the condition of at least 1 hour a day however, the change of its activity was found to be less than 12%. On the other hand, CAT repeatedly exposed to UV-A showed reduced size of the electrophoretic band which indicated a structure denaturation and its activity was significantly decreased. In the case of that the repeat exposure time was longer, CAT activity was completely lost even though some enzyme band was shown in the electrphoretic analysis. Conclusions: From these results, it was revealed that the degree and pattern in structural denaturation of antioxidative enzymes differently appeared according to the type of enzyme, and the degree of structural denaturation was not always consistent with the reduction in enzyme activity.

• Korean Journal of Microbiology
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• v.27 no.2
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• pp.85-91
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• 1989

The effect of SV40 and murine cytomegalovirus (MCMV) enhancers on the general and tissue-specific gene expression was investigated. Recombinant plasmids containing these transcriptional engancers downstream of a structural gene for chloramphenicol acetyl transferase (CAT) were constructed. The transient expression of CAT gene from these plasmids was measured in monkey (CV1PD) and HeLa cells. Both SV40 and MCMV engancers activated the expression of CAT gene by more than 20 and 150 folds, respectively, compared with engancerless plasmids. When the SV40 promoter, upstream of CAT gene, was replaced with 2.2 kbp of promoter regulatory region of bovine growth hormone (bGH) gene, there was no expression of CAT even in the presence of enhancers, reflecting the tissue-specific expression of bGH genes. However, when the bGH regulatory region was shortened to 230 bp, the expression level increased dramatically in the presence of SV40 enhancers. In contrast, the expression from the shortened promoter was only marginally activated by the stronger MCMV enhancer.

• Archives of Pharmacal Research
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• v.27 no.9
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• pp.968-972
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• 2004

A 1, 2-diglyceride-based multi-step colorimetric assay to measure the pancreatic lipase activ-ity was applied for the determination of the kinetic profiles of the lipase inhibition with a slight modification and the validity verification. With this assay method, our study revealed that platy-codin D, one of major constituents of Platycodi Radix, inhibits the pancreatic lipase activity in a competitive type, with the value of $K_1$ being 0.18${\pm}$0.02 mM. In addition, PO has affected the val-ues of $K_{m}$, app/ and $K_{cat}$/$K_{m}$ in a dose-dependent manner. The results shed a meaningful light on how PO mediates lipid metabolism in the intestinal tracts. On the other hand, since the revised assay is sensitive, rapid, and does not affect the accuracy to the kinetic properties, it is applica-ble not only to evaluation of the kinetic properties of the pancreatic lipase, but also to high-throughput screening of pancreatic lipase activity.

• Archives of Pharmacal Research
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• v.27 no.10
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• pp.1048-1052
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• 2004

A 1, 2-diglyceride-based multi-step colorimetric assay to measure the pancreatic lipase activity was applied for the determination of the kinetic profiles of the lipase inhibition with a slight modification and the validity verification. With this assay method, our study revealed that platycodin D, one of major constituents of Platycodi Radix, inhibits the pancreatic lipase activity in a competitive type, with the value of $K_I$ being 0.18${\pm}$0.02 mM. In addition, PD has affected the values of $K_{m,app}\;and\;K_{cat}/K_m$ in a dose- dependent manner. The results shed a meaningful light on how PD mediates lipid metabolism in the intestinal tracts. On the other hand, since the revised assay is sensitive, rapid, and does not affect the accuracy to the kinetic properties, it is applicable not only to evaluation of the kinetic properties of the pancreatic lipase, but also to highthroughput screening of pancreatic lipase activity.

• Korean Journal of Veterinary Service
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• v.45 no.4
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• pp.305-316
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• 2022

Bordetella (B.) bronchiseptica, Mycoplasma (M.) felis, and Chlamydia (C.) felis are considered as main bacterial pathogens of feline upper respiratory tract disease (URTD). In this study, a new triplex quantitative real-time polymerase chain reaction (tqPCR) assay was developed for the rapid and differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with the detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 1%. Based on the diagnostic results of the assay using 94 clinical samples obtained from cats with URTD signs, prevalence of B. bronchiseptica, M. felis, or C. felis was 10.6%, 36.2%, or 6.4%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported monoplex qPCR assays. The dual infection rates for B. bronchiseptica and M. felis or M. felis and C. felis was 2.1% or 3.2%, respectively. These results indicated that M. felis has been widely spread, and its co-infection with B. bronchiseptica or M. felis has been frequently occurred in Korean cat population. The developed tqPCR assay will serve as a promising tool for etiological and epidemiological studies of these three bacterial pathogens and the prevalence data obtained in this study will contribute to expanding knowledge about the epidemiology of feline URTD in Korea.

• Natural Product Sciences
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• v.6 no.1
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• pp.36-39
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• 2000

Acorenone, a diterpene isolated from Acorus gramineus, showed strong resistance inhibitory activity against multi-drug resistant microorganisms such as Staphylococcus aureus SA2, which has resistance to 10 usual antibiotics including chloramphenicol (Cm). At the level of $5\;{\mu}g/ml$ when combined with $50\;{\mu}g/ml$ of Cm. Bacterial resistance to Cm is due to the presence in resistant bacteria of an enzyme, chloramphenicol acetyltransferase (CAT), which catalyses the acetyl-CoA dependent acetylation of the antibiotic at C-3 hydroxyl group. To elucidate the mechanism of resistant inhibitory effect, the acorenone which had the strongest resistant inhibitory activity, was investigated on the CAT assay. As the result, the combination of Cm and acorenone showed the strongest inhibitory activity on CAT as noncompetitive and dose dependent manner.

• YAKHAK HOEJI
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• v.48 no.1
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• pp.93-98
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• 2004

The purpose of this study was to evaluate the effect of pharbitidis seed extract (PE) on antioxidant enzymes. The cytotoxicities of PE were measured by 3- (4,5-dimethlthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; The change of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activity assay were measured. The SOD activities by PE-treated groups were lower than control group's one. In the co-treated with hydrogen peroxide ($H_2O$$_2$) group, SOD activity was higher than $H_2O$$_2$ treated group's activity In the case of GPx, GPx activities were increased in both PE-treated and co-treated with $H_2O$$_2$ group. In the case of CAT $H_2O$$_2$ treated group's activityies were very increased. The CAT activities by PE-treated groups were lower than control group's one, but the activity of co-treated group with H $_2$O$_2$ was higher than that of control group's one. These results suggest that PE has antioxidant activity.