• 제목/요약/키워드: CAT assay

검색결과 112건 처리시간 0.03초

Expression of the VP2 protein of feline panleukopenia virus in insect cells and use thereof in a hemagglutination inhibition assay

  • Yang, Dong-Kun;Park, Yeseul;Park, Yu-Ri;Yoo, Jae Young;An, Sungjun;Park, Jungwon;Hyun, Bang-Hun
    • 대한수의학회지
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    • 제61권2호
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    • pp.19.1-19.7
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    • 2021
  • Feline panleukopenia virus (FPV) causes leukopenia and severe hemorrhagic diarrhea, killing 50% of naturally infected cats. Although intact FPV can serve as an antigen in the hemagglutination inhibition (HI) test, an accidental laboratory-mediated infection is concern. A non-infectious diagnostic reagent is required for the HI test. Here, we expressed the viral protein 2 (VP2) gene of the FPV strain currently prevalent in South Korea in a baculovirus expression system; VP2 protein was identified by an indirect immunofluorescence assay, electron microscopy (EM), Western blotting (WB), and a hemagglutination assay (HA). EM showed that the recombinant VP2 protein self-assembled to form virus-like particles. WB revealed that the recombinant VP2 was 65 kDa in size. The HA activity of the recombinant VP2 protein was very high at 1:215. A total of 143 cat serum samples were tested using FPV (HI-FPV test) and the recombinant VP2 protein (HI-VP2 test) as HI antigens. The sensitivity, specificity, and accuracy of the HI-VP2 test were 99.3%, 88.9%, and 99.3%, respectively, compared to the HI-FPV test. The HI-VP2 and HI-FPV results correlated significantly (r = 0.978). Thus, recombinant VP2 can substitute for intact FPV as the serological diagnostic reagent of the HI test for FPV.

고양이 말초혈액 다형핵백혈구의 유주활성에 있어서 계난백유래물질의 면역증강 효과 (Immunostimulating Effect of Chickent Egg White Derivatives on Chemotactic Activity of Feline Peripheral Blood Polymorphonuclear Cells)

  • 양만표;이재권
    • 한국임상수의학회지
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    • 제17권1호
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    • pp.21-27
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    • 2000
  • 고양이 말초혈액 다형핵백혈구 (PMN)의 유주활성에 있어서 계난백유래물질 (EWD)의 면역증강 효과를 검토하였다. PMN의 유주활성은 EWD 그 자체 및 EWD로 배양한 PMN의 배양상층액에서는 활성이 존재하지 않았다. 그러나 EWD로 배양한 MNC 배양상층액에서는 PMN에 대한 유주활성이 증가되었다. 본 활성은 checkerboard assay를 통해 진성 유주활성으로 밝혀졌다. 이러한 고양이 PMN 유주활성은 human recombinant interleukin-8(hr IL-8)에 의해서도 증가되었다. IL-8의 또 다른 특성인 세포 형태변형 실험에서도 EWD로 배양한 MNC의 배양상층액과 hr IL-8 모두에서 높은 수준의 세포 형태변형효과를 나타내었다. 이와 같이 EWD로 배양한 MNC의 배양상층액과 hr IL-8 모두에서 증가된 PMN의 유주활성 및 세포변형효과는 anti-human IL-8mAb에 의해 농도의존적으로 억제되었다. 이상의 결과로부터 EWD는 고양이 PMN의 유주활성에 있어서 면역증강 효과가 있으며, 이것은 EWD에의해 활성화된 MNC에서 분비되는 IL-8양 유주성 인자에 의해 PMN의 유주활성 증가 및 세포 형태 변형이 일어나는 것으로 사료되었다.

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고양이 말초혈액 다형핵백혈구의 유주성에 있어서 인삼 사포닌의 효과 (Effect of Ginseng Saponins on Chemotaxis of Feline Peripheral Blood Polymorphonuclear Cells)

  • 양만표;박세형;윤영원
    • 한국임상수의학회지
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    • 제15권1호
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    • pp.104-109
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    • 1998
  • 고양이 말초혈액 다형핵백혈구(PMNC)의 유주성에 있어서 인삼 saponins(ginseng total ssponin(GTS), ginseng PT saponin(GfT) 및 ginseng PD saponin(GPD))의 면역증강 효과를 검토하였다. PMNC에 대한 유주성을 hoyden chamber 변법으로 측정한 결과, GTS, GPT, GPD는 PMNC에 대해 직접적인 유주활성을 나타내지 않았다. 또한 인삼 saponins을 첨가하여 배양한 PMNC 배양상충액의 경우게는 미약한 유주활성을 나타내었다. 그러나 GPf및 GPD를 첨가하여 배양한 말초혈액 단핵구세포(MNC)배양상충액에서는 PMNC의 유주활성이 현저하게 증가하였다. PMNC에 대해 유주활성이 인정된 GPI)로 배양한 MNC 배양상충액을 이용하여 checkerboard assay를 실시한 결과 배양상충액의 농도차에 의존하는 진의 유주활성임을 알 수 있었다. 이상의 결과로부터 인삼 saponine 중 GPT 및 GPD가 고양 이 말초혈액 다형핵백혈구의 유주성을 중강시키는 효과가 있으며, 이것은 이들 saponins에 의해 활성화된 단핵구세포에서 분비되는 유주성인자에 의해 다형핵백혈구의 유주활성이 증 강되는 것으로 사료되었다.

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스트레스 유형에 따른 고양이 혈중 Cortisol농도의 변화 (Change of Blood Cortisol Concentration in Domestic Cats by Various Stress Types)

  • 나기정
    • 한국임상수의학회지
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    • 제15권1호
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    • pp.1-7
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    • 1998
  • The present study was performed to evaluate circadian rhythm of blood nortisol concentration and change of blood cortisol concentration by various stress type in domestic cats. The ELISA kit for cortisol measurement was evaluated on the effective assay range, day to day variation, within-run variation and accuracy. The results were 1-1,000 ng/ ml, 0.7-5.9%, 0.9-4.5% and 98%, respectively. The circadian rhythm of serum cortisol concentration is presented in cats. Change of serum cortisol concentration was also examined with several stressors, stimulation with hamster as a prey, beep electronic sounds, vehicle transportation and unfamiliar environment. Unfamiliar environment showed most remarkable change in serum cortisol level.

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광노화의 탄력소의 변화

  • 이규석
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 1996년도 4차 심포지움(Skin Biology Efficacy)
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    • pp.10-14
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    • 1996
  • 최근 오존층의 파괴로 인한 지구 자외선량의 증가로 피부암 및 광노화의 발생가능성이 증가되고있다. 광노화의 주요한 현상중의 하나는 탄력섬유상 물질이 축적되는 일광탄력 섬유증이다. 자외선은 탄력소 유전자의 전사과정증가를 유도하며 결국 비정상적인 탄력소의 증가를 유발한다는 보고가 있다. 본 연구에서는 피부 섬유아세포 및 각질형성세포를 배양하여 자외선이 탄력소 유전자발현에 미치는 영향 및 전사조절인자에 미치는 영향을 알아보고자 하였다. 피부섬유아세포에 UVB를 30mJ-200J/m 조사하여 nothern blotting한 결과 UV양이 증가함에 따라 탄력소 전사물이 증가되었고, 1001/$m^2$ 에서 최대의 증가를 보였다. 탄력소 promoter와 CAT을 접합시킨 pEP62 vector을 섬유아세포에 형질전환 시키고 UV에 의한 Promoter 활성을 본 결과 UV조사량이 증가할수록 활성이 증가되었고 200J/$m^2$ 에서 대조군에 비해 5배의 활성 증가를 보였다. 이 system에 광노화 억제물질로 알려진 retinoid를 5 $\times$ $10^{-6}$M처리하였을 때 UV에 의한 탄력소 promoter활성을 약 1/3로 감소시켰다. 각질 형성세포에서 UVB에 의한 transcription factor의 활성을 mobility shift assay에 의해 조사한 결과 AP-1 및 NFkB가 활성화됨을 볼 수 있었다.

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UVB 조사에 의한 육미지황탕의 광노화 피부 억제에 관한 실험적 연구 (Experimental Studies on the Inhibitory Effects of Yukmijiwhang-tang on Photoaging Skin Induced by UVB Irradiation)

  • 전혜숙;이창현;안홍석
    • 동의생리병리학회지
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    • 제28권5호
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    • pp.520-529
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    • 2014
  • The purpose of this study is to investigate the protective effects of Yukmijiwhang-tang(YM) water extracts against the UVB irradiation on the human keratinocyte HaCaT cells. We observed the effects of YM on the oxidative stress, gene expression of pro-inflammatory cytokine such as TNF-${\alpha}$ and IL-$1{\beta}$, and matrix metalloproteinase-9 in UVB-irradiated HaCaT cells. On the effects of oxidative stress and antioxidant function on the treatment with YM, The activity of xanthine oxidase(XO) was significantly decreased by treatment of YM in all the concentrations(p<0.01). The activity of superoxide dismutase(SOD) and catalase(CAT) was significantly increased by treatment of YM in a dose dependent manner(p<0.05 and p<0.01). DPPH radical was erased by treatment of YM under dose of $500{\mu}g/m{\ell}$ concentration. Treatment of HaCaT cells with YM had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(p<0.05, p<0.01, p<0.001). Gelatin zymography assay showed that YM downregulated the MMP-9 activity in UVB-irradiated HaCaT cells. RT-PCR analysis revealed that YM suppressed the expression of IL-$1{\beta}$ and MMP-9 however, it has no effects on the expression of TNF-${\alpha}$ and MMP-3. Our study suggests that Yukmijiwhang-tang exert protective actions on the UVB-irradiated HaCaT cells largely by anti-oxidative and anti-inflammatory processes.

제주도에(濟州道) 있어서 Toxoplasma 항체분포(抗體分布)에 관(關)한 연구(硏究) 1. 돼지, 고양이 및 식육취급자(食肉取扱者)에 있어서의 Toxoplasma 항체분포(抗體分布)에 대하여 (On the distribution of Toxoplasma antibodies in Cheju-Do 1. Distribution of Toxoplasma antibodies in swine, cats and butchers)

  • 김승호;김영주
    • 대한수의학회지
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    • 제29권3호
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    • pp.333-342
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    • 1989
  • Effect of raising types and environmental conditions on the infection of Toxoplasma in the swine, the cat and the man were studied in Cheju Island from Sept. 1987 to Aug. 1988. Blood samples were taken from 214 conventionally raised swine in 6 villages and 506 swine raised in swine specialized farms, 122 cats raised under free moving or restraned conditions in 8 locations, 113 butchers, and 210 villagers. Toxoplasma antibody values of the blood sera were determined using the enzymelinked immunosorbent assay (ELISA). The eating type of viscera was also investigated by using questionaires 1. When ELISA method was used, the percentage of Toxoplasma infected swine among the conventionally raised and of those raised in swine specialized farms were 60.7% and 21. 390, respectively. The respective mean of antibody values (${\pm}SD$) were 0.589 (${\pm}0.310$) and 0.385 (${\pm}0.237$) and differed very significantly (p<0.01). A significant difference was also found in antibody values among 6 villages (p<0.05). 2. The mean infection percentage of toxoplasma in the cat was 38.2%. the infection percentage for cats raised under free-moving and re~;trained condition were 37.0% and 38.2% respectively. The respective antibody values(${\pm}SD$) for toxoplasma were 0.600(${\pm}0.614$) and 0.637 (0.645), and did not difference significantly. 3. The infection percentage of toxoplasma in villagers and butchers were 26.2 and 38.3% respectively. The respective antibody values (SD) for toxoplasma were 0.429(${\pm}0.195$) and 0.341 (${\pm}0.236$), and differed very significantly (p<0.01). There were also highly significant differences Pyo-sun and other village (p<0.01). 4. Analysis of the questionaires showed that 26.0% of 392 villages eated liver and some villagers eated other viscera.

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Germination and Biochemical Changes in Accelerated Aged and Osmoprimed Pinus thunbergii Seeds

  • Kim, Du-Hyun;Han, Sim-Hee;Lee, Jae-Cheon
    • 한국산림과학회지
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    • 제99권2호
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    • pp.244-250
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    • 2010
  • The aim of this study was to investigate relationship among seed viability and enzymes activities involved in scavenging reactive oxygen species (ROS), especially, superoxide dismutase (SOD), glutathione reductase (GR), and catalase (CAT). In other respects, osmopriming has been demonstrated to reinvigorate aged seeds. Various viabilities of seeds that were ranged from 80 to 100% of germination rate could be produced using osmopriming and accelerated aging treatments. Priming treatment of Pinus thunbergii seeds for 3 days at $15^{\circ}C$ with a polyethylene glycol solution at -1.2 MPa improved their subsequent germination at $25^{\circ}C$. Accelerated aging (3, 6, 9, and 12 days at $41^{\circ}C$ and 100% relative humidity) decreased seed germination percentage depending on aging treatment duration. Electrolyte conductivities of seeds were measured as assay of membrane integrity. The conductivity from electrolyte leakage of P. thunbergii seed was also correlated with seed germinability. Conductivity for control seeds that had 95% of germination percentage was 3.48 ${\mu}S\;g^{-1}$, but jumped as doubled (7.98 ${\mu}S\;g^{-1}$) in 12-day-aged seed that had 80% of germination percentage. Our results demonstrate that aging of P. thunbergii seeds is associated with changes in the electrolyte leakage, lipid peroxidation, and antioxidant defense system. Priming of aged seeds progressively restored the initial germinative ability and resulted in a marked decrease in the levels of MDA and conductivity of seed leachate. These effects of priming were also well recovered of GR and CAT activities in aged seed. The improved seed quality by priming treatment appears at least partly attributable to reduced lipid peroxidation, resulting from enhanced antioxidative enzyme activities that are suggesting the antioxidant defense systems play a key role in seed vigor.

A fluorogenic method for measuring enteropeptidase activity: spectral shift in the emission of GD4K-conjugated 7-amino-4-methylcoumarin

  • Choi, Mal-Gi;Lee, Eung-Yeong;Chung, Hye-Shin;Jang, Sei-Heon;Lee, Chang-Woo
    • BMB Reports
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    • 제44권7호
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    • pp.458-461
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    • 2011
  • Enteropeptidase is a serine protease secreted by the pancreas and converts inactive trypsinogen to active trypsin. Enteropeptidase cleaves the C-terminal end of the substrate recognition sequence Asp-Asp-Asp-Asp-Lys ($D_4K$). The assay for enteropeptidase has utilized $GD_4K$-conjugated 2-naphthylamine ($GD_4K$-NA) as a fluorogenic probe over the last 30 years. However, no other $D_4K$-conjugated fluorogenic substrates of enteropeptidase have been reported. Furthermore, naphthalene is known as carcinogenic to humans. In this study, we used shift in the emission spectrum of $GD_4K$-conjugated 7-amino-4-methylcoumarin ($GD_4K$-AMC) as a fluorogenic method to measure enteropeptidase activity. The kinetic analysis revealed that enteropeptidase has a $K_M$ of 0.025 mM and a $k_{cat}$ of 65 $sec^{-1}$ for $GD_4K$-AMC, whereas it has a $K_M$ of 0.5 to 0.6 mM and a $k_{cat}$ of 25 $sec^{-1}$ for $GD_4K$-NA. The optimum pH of $GD_4K$-AMC hydrolysis was pH 8.0. Our data indicate that $GD_4K$-AMC is more suitable as a substrate for enteropeptidase than $GD_4K$-NA.

Efficacy of Ronidazole for Treatment of Cats Experimentally Infected with a Korean Isolate of Tritrichomonas foetus

  • Lim, Sun;Park, Sang-Ik;Ahn, Kyu-Sung;Oh, Dae-Sung;Shin, Sung-Shik
    • Parasites, Hosts and Diseases
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    • 제50권2호
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    • pp.161-164
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    • 2012
  • To evaluate the efficacy of ronidazole for treatment of Tritrichomonas foetus infection, 6 Tritrichomonas-free kittens were experimentally infected with a Korean isolate of T. foetus. The experimental infection was confirmed by direct microscopy, culture, and single-tube nested PCR, and all cats demonstrated trophozoites of T. foetus by day 20 post-infection in the feces. From day 30 after the experimentally induced infection, 3 cats were treated with ronidazole (50 mg/kg twice a day for 14 days) and 3 other cats received placebo. Feces from each cat were tested for the presence of T. foetus by direct smear and culture of rectal swab samples using modified Diamond's medium once a week for 4 weeks. To confirm the culture results, the presence of T. foetus rRNA gene was determined by single-tube nested PCR assay. All 3 cats in the treatment group receiving ronidazole showed negative results for T. foetus infection during 2 weeks of treatment and 4 weeks follow-up by all detection methods used in this study. In contrast, rectal swab samples from cats in the control group were positive for T. foetus continuously throughout the study. The present study indicates that ronidazole is also effective to treat cats infected experimentally with a Korean isolate of T. foetus at a dose of 50 mg/kg twice a day for 14 days.