• Archives of Pharmacal Research
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• 제27권10호
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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.

• 생명과학회지
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• 제19권4호
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• pp.442-448
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• 2009

대부분의 열대 식물은 chilling에 민감하게 반응한다. 대표적 열대 식물인 벼 잎에 대한 light-chilling 처리와 이 후의 회복기(post-chilling) 동안 일어나는 반응들을 알아보았다. Chilling 시 벼 잎에서의 광합성 효율($F_v/F_m$)은 대조구보다 50% 감소하였고, 상대적으로 $H_2O_2$ 양은 48% 증가하였다. 항산화 효소들 중 SOD와 GR 활성은 chilling과 post-chilling 시 증가하였다. 특히 SOD isoforms의 경우 CuZn-SOD와 Mn-SOD 가 발현된 반면 Fe-SOD는 발현되지 않았다. CAT 활성은 chilling 시 감소하였으며, 반면에 APX는 크게 증가하였다. Chilling 시 CAT의 isoforms의 변화를 보면, CAT-2와 -3의 활성이 감소한 것과 대조적으로 post-chilling 시 이들 isoforms의 활성은 증가하였다. 이처럼 APX와 CAT 활성은 벼 잎이 chilling stress를 겪게 될 때 상반되는 변화를 보여주었다.

• 공업화학
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• 제2권2호
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• pp.108-118
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• 1991

황화 $CoMo/{\gamma}-Al_2O_3$ 촉매를 사용하여 473~723K의 온도와 $10{\sim}15{\times}10^5Pa$의 압력 그리고 접촉시간 0.0125~0.03 g-cat hr/ml-feed 범위에서 pyridin의 수첨탈질반응과 m-cresol의 수첨탈산소 반응의 상호작용 및 그 속도론에 관하여 연구하였다. Pyridin의 수첨탈질반응과 m-cresol의 소첨탈산소 반응의 상호반응을 저지억제하였으며, pyridin에 의한 m-cresol의 수첨탈산소반응의 억제효과는 m-cresol에 의한 pyridin의 수첨탈질반응의 억제효과보다 더 컸으나 반응성은 m-cresol이 더 높았다. Pyridin의 수첨탈질반응 속도식 및 m-cresol의 수첨탈산소반응 속도식을 LHHW 모델을 이용하여 구한 결과 ${\gamma}_{HDN}=k_{HDN}{\cdot}K_pC_p/(1+K_cC_c+K_pC_p)$, ${\gamma}_{HDO}=k_{HDO}{\cdot}K_cC_c/(1+K_cC_c+K_pC_p)$였다. 각 온도에서 반응속도상수 및 흡착평형상수를 구하여 Arrhenius plot 과 Van't Hoff plot을 행하여 구한 활성화 에너지값은 pyridin과 m-cresol이 각각 16.21 Kcal/mole, 13.83 Kcal/mole이었고, 흡착열은 각각 -6.458 Kcal/mole, -5.045 Kcal/mole이었다.

• 한국동물위생학회지
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• 제38권1호
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• pp.31-36
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• 2015

Two cats were obtained from a cat kennel. Over the previous 7 days, the cats had shown cough, anorexia, depression and nasal discharge. In this study, the consensus PCR was able to detect successfully Mycoplasma species in nasal swab samples of the cats. To identify feline mycoplasma species from the lung tissue of the cats with pneumonia, Mycoplasma species-specific PCR reactions were conducted. As the results, we could identify M. felis by the positive amplified DNAs. On the other hand, we could not detect any positive reactions with the PCR reaction for M. arginini, M. canis, M. edwardii, M. cynos, M. gateae, M. maculosum, M. molared, M. opalescens, M. spumans and Mycoplasma HRC-689. In conclusion, we detected M. felis from the kenneled cats with pneumonia. We suggested that this consensus PCR would be useful and effective for monitoring Mycoplasma species in various kinds of animals including cats. The application of preceding consensus PCR before the species-specific PCRs may be the most recommended strategy for the identification of Mycoplasma spp.

• Applied Microscopy
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• 제22권1호
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• pp.42-54
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• 1992

To understand the structural changes of the myocardial myocytes and endothelial cells in ischemic and reperfused heart, and to elucidate their roles in those conditions, the authors observed cat and rat myocardium ultrastructurally and evaluated them with morphometric techniques. In cat, mild ischemia and moderate degree reperfusion injury was induced by ligation of the anterior interventricular branch of left coronary artery and reperfusion. In rat, severe ischemia and irreversible reperfusion iniury was made using in vitro Langendorff techniques. In normal cat myocytes, the volume densities of cytoplasm, myofibrils, mitochondria, sarcoplasmic reticulum and T tubules were $0.11{\pm}0.013,\;0.51{\pm}0.096,\;0.25{\pm}0.082,\;0.09{\pm}0.008,\;0.02{\pm}0.010$ (Mean${\pm}$S.D.) respectively, and the myofibril/mitochondria ratio was $2.33{\pm}1.379$. The numerical density and average volume of mitochondria were $0.76{\pm}0.210/{\mu}m^3$ and $0.33{\pm}0.057{\mu}m^3$ respectively. In normal cat endothelial cells, the volume densities of cytoplasm, cytoplasmic vesicles, tubular systems (including endoplasmic reticulum and Golgi apparatus) and mitochondria were $0.43{\pm}0.023,\;0.28{\pm}0.007,\;0.22{\pm}0.021,\;0.03{\pm}0.014$ respectively. The mean thickness of endothelial cells was $230{\pm}45.2{\mu}m$. The numerical density and average volume of cytoplasmic vesicles were $508{\pm}55.0/{\mu}m^3,\;578{\pm}104.8nm^3$ respectively. In cat myocytes which received mild ischemic injury, the volume densities of organelles were not changed significantly in ischemic and reperfusion states. In reperfusion group myocytes, the numerical density of mitochondria was decreased significantly and the average volume was increased significantly. In endothelial cells, the volume density of tubular system in ischemic group and the average volume of cytoplasmic vesicles in reperfusion group were increased significantly. In rat myocytes which received severe ischemic injury, the volume density and average volume of mitochondria were increased significantly, and the volume density of sarcoplasmic reticulum and numerical density of mitochondria were decreased significantly in both ischemic and reperfusion groups. In ischemic and reperfused endothelial cells, the volume density and numerical density of cytoplasmic vesicles, the volume density of cytoplasm were decreased significantly. The volume densities of tubular system were increased significantly in both ischemic and reperfused groups. The volume density of mitochondria in ischemic group and the average volume of cytoplasmic vesicles in reperfusion group showed significant increase. The authors, based on the above observations, conclude that the mitochondria of myocytes and the cytoplasmic vesicles of endothelia are the first group of targets in ischemic and reperfusion injury and in this respect, the degree of ischemic insult is not significant. The role of myocyte mitochondria in reperfusion injury may be insignificant, but endothelial cells may contribute actively to reperfusion injury.

• 한국작물학회지
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• 제47권5호
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• pp.341-351
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• 2002

In order to examine the mechanistic basis for differential sensitivities to chilling and subsequent recovery between two rice (Oryza sativa L.) cutivars, a chilling-tolerant japonica type (Ilpumbyeo) and a chilling-susceptible indica type (Taebaekbyeo), changes of physiological responses and antioxidant enzymes were investigated. Both cultivars at 3 leaf stage were exposed at a low temperature of $5^{\circ}C$ for 3 days and subsequently recovered in a growth chamber at a $25^{\circ}C$ for 5 days with 250 mmol $m^{-2}$ $s^{-1}$. Physiological parameters such as leaf fresh weight, relative water content, cellular leakage, lipid peroxidation, and chlorophyll a fluorescence showed that the chilling tolerant cultivar had a high tolerance during chilling. However, the chilling-susceptible cultivar revealed severe chilling damages. The chilling-tolerant cultivar was also faster in recovery than the chilling-susceptible cultivar in all parameters examined. We analyzed the activity and isozyme profiles of four antioxidant enzymes which are: superoxide dismutase (SOD), caltalase (CAT), ascorbate peroxidase (APX), and glutation reductase (GR). We observed that chilling-tolerance was due to a result of the induced or higher antioxidant enzyme system, CAT and APX in leaves and SOD, CAT, APX, and GR in roots. Especially, we observed the most significant differences between the chilling-tolerant cultivar and -susceptible cultivar in CAT and APX activity. Also in isozyme profiles, CAT and APX band intensity in the chilling-tolerant cultivar was distinctively higher than in the chilling-susceptible cultivars during chilling and recovery. Thus, the cold stability of CAT and APX are expected to contribute to a tolerance mechanism of chilling in rice plants. In addition, the antioxidative enzymes activity in roots may be more important than in that of leaves to protect chilling damage on rice plants.

• Bulletin of the Korean Chemical Society
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• 제34권9호
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• pp.2671-2674
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• 2013

To elucidate the role of the evolutionarily conserved Tyr8 residue in rice Phi-class GSTF3, this amino acid was replaced with alanine and phenylalanine by site-directed mutagenesis, respectively. The replacement of Tyr8 with Ala significantly affected the catalytic activity and the kinetic parameters, whereas the substitutions of Tyr8 with Phe had almost no effect. The Y8A mutant resulted in approximately 90-100% decrease of the specific activity. Moreover, the Y8A mutant resulted approximately in 2-fold increase of $K_m$, approximately 60-80% decrease of $k_{cat}$, and approximately 6.5-fold decrease in $k_{cat}/K_m$. From the pH/log $k_{cat}/K_m$ plot, $pK_a$ values of the GSH in the wild-type enzyme-GSH complex, Y8A-GSH complex and Y8F-GSH complex were estimated to be approximately 6.8, 8.5 and 6.9, respectively. From these results, we suggest that the evolutionarily conserved Tyr8 residue in OsGSTF3 seems to influence the structural stability of the active site of OsGSTF3 rather than directly its catalytic activity.

• BMB Reports
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• 제34권5호
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• pp.472-477
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• 2001

Ornithine decarboxylase (ODC, EC 4.1.1.17) is the first and key enzyme in eukaryotic polyamine biosynthesis. The cDNA encoding ornithine decarboxylase from Nicotiana glutinosa was cloned ($GeBank^{TM}$ AF 323910) and expressed in E. coli. Site directed mutagenesis were performed on several highly conserved cysteine residues. Among the mutants, C115A showed significant changes in the kinetic properties. The $K_m$ value of the C115A mutant was $1790\;{\mu}M$, which was 3-fold higher than that of the wild-type ODC. There was a dramatic decrease in the $k_{cat}$, values of the C115A mutant, compared to that of the wild-type ODC, which had a $k_{cat}$ value of $77.75\;s^{-1}$. C115A caused a shift in the optimal pH from 8.0 to 8.4. Considering these results, we suggest that cys-115 is involved in the catalytic activity of N. glutinosa ODC.

• The Korean Journal of Physiology
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• 제14권2호
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• pp.21-28
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• 1980

In order to investigate the effect of cholinergic substance on the electrical and the mechanical activities of the stomach muscle, 10 isolated cat stomachs were studied. At various sites of a stomach muscle preparation, the electrical activity was monopolarly recorded by using capillary electrodes containing chlorided silver wires, and the isometric contractile activity was recorded simultaneously at the terminal portion of the antrum in Krebs solution$(36^{\circ}C)$ which was aerated with a gas mixture consisting of 95% $O_2$ and 5% $CO_2$. The recording of these activities were performed before (control period) and after acetylcholine$(10^{-5}M)$ and atropine $(10^{-6}M)$ administrations serially. Following results were obtained: 1) The mean frequency of the slow wave was $4.36{\pm}0.22\;cycles/min$ at all the various sites of the cat stomach. The slow wave was propagated caudad in sequence and its velosity of propagation increased as the slow wave approached the pylorus in normal Krebs solution. 2) After acetylcholine administration, the frequency of the slow wave increased transiently and the increase of slow wave frequency was followed by the isometric contraction of antral muscle in association with the second potential which succeeded the slow wave. 3) By atropine administration, the stimulatory effect of acetylcholine on the antral muscle contraction was abolished completely, and the frequency of the slow wave decreased significantly compared with that of the control period, which tendency was more prominent in the antrum. The above results suggest that the transient increase in the frequency of gastric slow wave by acetylcholine may have some influence upon the contraction mechanism of the cat antral muscle.

• Archives of Pharmacal Research
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• 제10권2호
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• pp.80-87
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• 1987

To get a better insight into the exxistence and the role of a Na-Ca exchange mechanism in smooth muscle, the effect of Na substitution with sucrose on tension development, cellular Ca uptake and $^{45}Ca$ efflux was investigated using isolated cat ileal longitudinal muscle strips. Experimental results were summarized as follows;1) Exposure of the cat ileal longitudinal muscle to Na-free solution induced a contraction, and the magnitude of the contraction increased after incubation of the muscle strips with ouabain ($2{\times10^{-}5}$M) for 1hr. 2) Cellular Ca uptake in Na-free solution increased with an increase in Na content of the Na-loading media, and a linear relationship existed between tissue Na content and cellular Ca uptake for 10 min 3) After tissues were equilibrated in PSS containing $^{45}Ca$ for 2hr, cellular Ca uptake decreased with rising the external Na concentration. 4)Removal of medium Na or inhibition of the Na-K pump decreased the rate of $^{45}Ca$ efflux. These results strongly suggested that Na substitution increases cellular Ca uptake and decreases the rate of $^{45}Ca$ efflux via a Na-Ca exchange mechanism.