• Parasites, Hosts and Diseases
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• v.54 no.2
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• pp.239-241
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• 2016

Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

• The Journal of Internal Korean Medicine
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• v.28 no.1
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• pp.166-175
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• 2007

Objectives : This study was designed to evaluate the neuroprotective effect of Cirsium japonicum and Silibinin on lipopolysaccharide-induced inflammation in BV2 microglial cells. Methods : We studied on the neuroprotective effect of lipopolysaccharide-induced inflammation using MTS assay, western blot, and nitric oxide detection on mouse BV2 microglial cells. Results : Cirsium japonicum dose-dependently (50${\mu}g/ml$${\sim}$$250{\mu}g/ml$) inhibited nitrite production and iNOS expression in lipopolysaccharide-induced BV2 microglia and also significantly reduced lipopolysaccharide-induced COX-2 activation in western blot. Silibinin dose-dependently (10${\mu}M$${\sim}$$100{\mu}M$) inhibited nitrite production and iNOS expression in lipopolysaccharide-induced BV2 microglial cells. Silibinin also significantly reduced lipopolysaccharide-induced COX-2 activation in western blot. Conclusion : These effects of neuroprotection related to anti-inflammation suggest that Cirsium japonicum and Silibininmay be useful candidates for the development of a drug for related neurodegenerative diseases.

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.9-15
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• 1998

To investigate the involvement of protein kinase C(PKC) isoenzyme in the testes which control spermatogenesis and hormone secretion, we examined cellular distribution of four types of PKC $\alpha$, ${\beta}I$, ${\delta}$ and ${\theta}$ in the horse testes using PKC antisera by western blot analysis and immunohistochemistry. By the western blot analysis, PKC $\alpha$ and ${\beta}I$ were detected at 82KD, while PKC ${\delta}$ and ${\theta}$ were detected at 80KD in the testes of both juvenile and adult horses. In juvenile horse, PKC $\alpha$, ${\delta}$ and ${\theta}$ except ${\beta}I$ were not detected in the cells of the testes, whereas PKC ${\beta}I$ was immunoreacted with only in spermatocytes. In adult, PKC $\alpha$, ${\beta}I$, ${\delta}$ and ${\theta}$isoenzymes were localized in interstitial cells of the testes. In the seminiferous tubules, PKC ${\beta}I$ is localized in spermatocyte, spermatid and spermatozoa, while PKC ${\delta}$ is localized only in spermatids. We suggest that this is a first report to localize PKC in the testes of horse and PKC isoenzymes are upregulated in the cells of horse testes depending on ages. These findings also suggest that certain PKC isoenzyme plays an important role in the signal transduction of spermatogenic cells and interstitial cells in horse testes.

• Korean Circulation Journal
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• v.48 no.12
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• pp.1135-1144
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• 2018

Background and Objectives: Mitochondria play a key role in the pathophysiology of heart failure and mitochondrial permeability transition pore (MPTP) play a critical role in cell death and a critical target for cardioprotection. The aim of this study was to evaluate the protective effects of cyclosporine A (CsA), one of MPTP blockers, and morphological changes of mitochondria and MPTP related proteins in monocrotaline (MCT) induced pulmonary arterial hypertension (PAH). Methods: Eight weeks old Sprague-Dawley rats were randomized to control, MCT (60 mg/kg) and MCT plus CsA (10 mg/kg/day) treatment groups. Four weeks later, right ventricular hypertrophy (RVH) and morphological changes of right ventricle (RV) were done. Western blot and reverse transcription polymerase chain reaction (RT-PCR) for MPTP related protein were performed. Results: In electron microscopy, CsA treatment prevented MCT-induced mitochondrial disruption of RV. RVH was significantly increased in MCT group compared to that of the controls but RVH was more increased with CsA treatment. Thickened medial wall thickness of pulmonary arteriole in PAH was not changed after CsA treatment. In western blot, caspase-3 was significantly increased in MCT group, and was attenuated in CsA treatment. There were no significant differences in voltage-dependent anion channel, adenine nucleotide translocator 1 and cyclophilin D expression in western blot and RT-PCR between the 3 groups. Conclusions: CsA reduces MCT induced RV mitochondrial damage. Although, MPTP blocking does not reverse pulmonary pathology, it may reduce RV dysfunction in PAH. The results suggest that it could serve as an adjunctive therapy to PAH treatment.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.95-98
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• 2000

An immunochemical method has been develooped as the most sensitive tool for studying the expression of quinoproteins containing pyrroloquinoline qinone(PQQ) in E. coli. The PQQ was conjugated to bovine serum albumin (BSA), and the conjugant was purified by using a $KwikSep^{TM}$ dextran desalting column chromatography. The PQQ-BSA conjugant was immunized to rabbits, and the IgG fractions of the antisera were purified. The most sensitive antibody against PQQ-BSA conjugant recognized some nanogram quantity of the antigen on the blot, but had little cross reactivity with BSA. Using this batch of the antibody, all the immunochemical assays of quinoproteins in E. coli were preformed. Some six different PQQ-specfic spots were detected by Western blot analysis of the soluble proteins in E. coli were performed. Some six different PQQ-specific spots were detected by Western blot analysis of the soluble proteins in E. coli after two-dimensional gel electrophoresis. Their molecular weights on the blot were estimated to be about 100-, 90-, 72-, 58-, 52-, and 50kDa. Their pI values fell in the range from 4.8 to 5.5. These results stronly suggest that quinoproteins are present in E. coli, and that the protein moieties were covalently bound to PQQ.

• Journal of Life Science
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• v.17 no.10
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• pp.1321-1329
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• 2007

Deer antler tissue contains the most rapidly growing bone in the animal kingdom. Thus, it is likely that growing antler tissue is a rich source of local paracrine bone-stimulating factors. Growth factors, at least the insulin-like growth factor (IGF), control the bone-remodelling process. In this study, we tried to isolate and purify IGF-I from fresh antler tissue by the routine isolation and purification of protein. The purification involved ammonium sulfate precipitation, DEAE-Sepharose CL-60 ion-exchange chromatography, CM-Sepharose CL-6B ion-exchange chromatography, and Sephadex G-50 chromatography. Purified fractions from each step were analyzed by high-performance liquid chromatography (HPLC), SDS polyacrylamide gel electrophoresis (SDS-PACE), Dot-blot, and Western-blot methods. Furthermore, the quantification of partially purified IGF-I was calculated by enzyme-linked immunosorbent assays (ELISA) using antibody to human recombinant IGF-1. SDS-PAGE analysis of the final fraction yielded two molecular bands and the signal band was at 12 kDa on the Western-blot film. This purified IGF-I fraction showed a peak at retention time of eight min. The quantity of IGF-I in 20 g deer antler tissue as starting weight was calculated using a standard curve to be 2910 ng/ml, and total IGF-I amount is 0.291 g. The results show that IGF-I, which can be found in deer antler, can be partially purified and quantified by classic protein isolation methods.

• Journal of Life Science
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• v.24 no.2
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• pp.186-190
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• 2014

Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

• Parasites, Hosts and Diseases
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• v.36 no.4
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• pp.269-275
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• 1998

Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7％) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG 1), and 22 kDa （SAG2) bands. With N. caninum antigen, the number of reactive bands was reduced, however a 43 kDa band reacted in three cases in Toxoplasma-positive sera in addition to one in Toxoplasma-negative control sera. All sera of the Toxoplasma-positive group labeled surface membrane of T. gondii, but reacted differently with N. caninum. Fluorescence was detected in surface membrane, subcellular organelles, or both in N. caninum. And one case in the Toxoplasma-negative group also reacted with N. caninum strongly in subcellular organelles. This suggested that the antibody against N. caninum may be present in human sera although the positive rate was very low in this study. The possibility of human infection with N. caninum remains to be evaluated further.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.269-269
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• 1994

GABA shunt에 관여하는 두 가지 효소인, GABA transminase와 Succinic semialdehyde reductase에 대한 단일 클론 항체를 생산하고 이들 항체의 특성을 살펴보았다. 소의 뇌에서 순수 정제된 효소를 동물에 주사한 후 immunodot-blot 분석법에 의하여 일차적으로 항체를 분비하는 hybridoma를 골라낸 후 생산된 단일 클론 항체가 특이적으로 이들 효소와 반응하는 가를 알아보기 위하여 Western blot 분석을 실시하였다. 뇌조직에서 추출한 총단백질을 SDS 전기영동법에 의하여 분리한 후 이들 항체를 처리한 결과, GABA-T에 대한 항체는 특이적으로 분자량이 50kDa에 해당하는 단백질 밴드만을 인지하였고 SSA reductase에 대한 항체의 경우 분자량이 34kDa 크기의 단백질 밴드와 반응하였다. 이들 분자량은 순수 정제된 소뇌의 효소 단백질의 크기에 해당하는 것을 확인하였다. 이들 소뇌의 효소에 대한 항체를 추적물체로 사용하여 다른 포유동물자 조류의 효소와 비교하는 cross-reactivity 연구를 수행하였다. 소, 돼지, 토끼, 쥐 (rat), 개, 고양이 그리고 닭의 뇌를 제거한 후 총단백질을 추출하고 Western blot을 해본 결과 GABA transaminase의 경우 조류를 제외하고 다른 포유동물에서는 같은 분자량의 단일 단백질 밴드를 확인하였고 SSA reductase의 경우 조류를 포함하는 모든 동물에서 같은 분자량의 밴드를 확인하였다 이상의 결과로 미루어 보아 포유류 뇌에 있는 이들 효소들은 변역학적으로 아주 유사하다고 사료된다.

• Biomolecules & Therapeutics
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• v.8 no.2
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• pp.107-112
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• 2000

To investigate the alteration of airway mucin in airway disease patients, immunoassay procedures were employed using monoclonal antibodies HM02 and HM03 (Hybridoma, 18,457-463, 1999). Alteration of mucin release was determined by ELISA and the integrity of mucin was determined by Western blot. In ELISA, it was found that mucin release increased from pneumonia, chronic cough, bronchiectasis, eosinophilic pneumonia, lung cancer and bronchial asthma patients. In Western blot, the increase in immunoreactivity was observed in case of pneumonia, chronic cough, bronchiectasis and bronchial asthma. In bronchial asthma, there was no obvious degradation of mucin while in other diseases, varying degree of mucin degradation was observed. The data from the present study implicate that HMO2 and HM03 are suitable for the immunological analysis of mucin in airway disease patients. The role of increased mucin release and varying degree of mucin degradation on airway diseases should be further investigated in the future.