• Title/Summary/Keyword: K_{cat}/K_m$

Search Result 396, Processing Time 0.038 seconds

Cellular Pathways in Agonist-induced Gallbladder Muscle Contraction in the Cat (고양이의 담낭근 수축에 있어서 세포내 기전)

  • Rhim, Byung-Yong;Kim, Chi-Dae;Kim, Dong-Heon;Biancani, Piero;Behar, Jose
    • The Korean Journal of Pharmacology
    • /
    • v.32 no.1
    • /
    • pp.67-74
    • /
    • 1996
  • Cholecystokinin octapeptide (CCK-8), acetylcholine (ACh) and KCl caused a dose dependent contraction in muscle cells enzymatically digested from cat gallbladder. Maximal contraction was obtained at concentration of $10^{-9}M$ for CCK-8, $10^{-5}M$ for ACh and 20mM for KCl. CCK-8 induced contraction was unaffected in calcium free physiological salt solution (PSS) and was completely blocked by strontium substitution for calcium (p<0.001). In contrast, KCl evoked contraction was blocked in calcium free PSS (p<0.01) but was unaffected by strontium replacement of calcium. The contraction elicited by ACh was only slightly reduced in calcium free PSS (p<0.05) and was unaltered by strontium. Muscle cells permeabilized with saponin contracted in response to inositol 1,4.5-trisphosphate $(IP_3)$ and CCK-8. The contraction was blocked by the calmodulin antagonist CGS 9343B (p<0.001), whereas heparin completely blocked the effect of $IP_3$ (p<0.001). The protein kinase C (PKC) antagonist H7 had no effect on either agonist. We conclude that CCK-8 induced gallbladder muscle contraction is mediated by $IP_3$ dependent intracellular calcium release from intracellular stores and a calmodulin dependent pathway; ACh may utilize both extracellular and intracellular calcium. KCl causes muscle contracrion through influx of extracellular calcium and a calmodulin independent machanism.

  • PDF

Application of PIT Tag and Radio Telemetry Research Methods for the Effective Management of Reptiles in Korea National Parks (국립공원 내 파충류 자원의 효율적인 관리를 위한 PIT (passive integrated transponder) tag과 무선추적방법(radio telemetry)의 적용)

  • Lee, Jung-Hyun;Lee, Heon-Joo;Ra, Nam-Yong;Kim, Ja-Kyeong;Eom, Jun-Ho;Park, Dae-Sik
    • Korean Journal of Environmental Biology
    • /
    • v.27 no.2
    • /
    • pp.146-154
    • /
    • 2009
  • To suggest effective ways of the management of reptiles in Korea national parks, we applied PIT (passive integrated transponder) tag and radio telemetry research methods in the monitoring of reptiles in Odaesan and Woraksan national parks. We implanted PIT tags into 137 snakes (six species) captured in both the national parks between March 2006 and October 2008 and also radio-tracked two Korean rat snakes (Elaphe schrenckii anomala) from September 2007 to November 2008 in the Woraksan national park. Of total 137 snakes which we had inserted the PIT tag, 17 snakes (12.40%) were recaptured. Based on the PIT tag data of the recapture, we successfully obtained the annual growth rate of Korean cat snakes (Elaphe dione) and Red-tongue pit-viper (Gloydius ussuriensis). Home range of the Korean rat snakes based on the data of radiotracking was estimated as 389,600$m^2$ (MCP: Minimum convex polygon) and 471,800$m^2$ (Kernel 95%) for males and 162,500$m^2$ and 208,700$m^2$ for females. These results suggest that if we apply PIT tag and radio telemetry research methods to manage reptiles in Korea national parks, it could greatly increase our understanding about their basic ecology and as the result, it could allow us to develop better management and conservation ways of reptiles in Korea national parks.

Gene Cloning, Expression, and Characterization of a $\beta$-Agarase, AgaB34, from Agarivorans albus YKW-34

  • Fu, Xiao Ting;Pan, Cheol-Ho;Lin, Hong;Kim, Sang-Moo
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.3
    • /
    • pp.257-264
    • /
    • 2009
  • A $\beta$-agarase gene, agaB34, was functionally cloned from the genomic DNA of a marine bacterium, Agarivorans albus YKW-34. The open reading frame of agaB34 consisted of 1,362 bp encoding 453 amino acids. The deduced amino acid sequence, consisting of a typical N-terminal signal peptide followed by a catalytic domain of glycoside hydrolase family 16 (GH-16) and a carbohydrate-binding module (CBM), showed 37-86% identity to those of agarases belonging to family GH-16. The recombinant enzyme (rAgaB34) with a molecular mass of 49 kDa was produced extracellularly using Escherichia coli $DH5{\alpha}$ as a host. The purified rAgaB34 was a $\beta$-agarase yielding neoagarotetraose (NA4) as the main product. It acted on neoagarohexaose to produce NA4 and neoagarobiose, but it could not further degrade NA4. The maximal activity of rAgaB34 was observed at $30^{\circ}C$ and pH 7.0. It was stable over pH 5.0-9.0 and at temperatures up to $50^{\circ}C$. Its specific activity and $k_{cat}/K_m$ value for agarose were 242 U/mg and $1.7{\times}10^6/sM$, respectively. The activity of rAgaB34 was not affected by metal ions commonly existing in seawater. It was resistant to chelating reagents (EDTA, EGTA), reducing reagents (DTT, $\beta$-mercaptoethanol), and denaturing reagents (SDS and urea). The E. coli cell harboring the pUC18-derived agarase expression vector was able to efficiently excrete agarase into the culture medium. Hence, this expression system might be used to express secretory proteins.

Protective Effect of PineXol® on Hydrogen Peroxide-induced Apoptosis on SK-N-MC Cells and Focal Ischemia Rodent Models (파인엑솔이 과산화수소로 유도된 SK-N-MC 세포와 뇌졸중 백서 모델에서의 보호효과)

  • Hong, Soon-O;Han, Kyung-Hoon;Lee, Seung-Hee;Kim, Doh-Hee;Song, Kwan-Young;Han, Sung-Hee
    • The Korean Journal of Food And Nutrition
    • /
    • v.29 no.6
    • /
    • pp.923-929
    • /
    • 2016
  • The purpose of this study was to evaluate the protective effect of $PineXol^{(R)}$ on $H_2O_2$-induced cell death in SK-N-MC cells, and in early stage focal ischemia rodent model. SK-N-MC cells were pre-treated with $200{\mu}M$ $H_2O_2$ or various concentrations of $PineXol^{(R)}$ (10, 30, and 50 pg/mL) for 24 h, and then exposed to $H_2O_2$ for 3 h. Cell death was assessed by the CCK-8 assay, reactive oxygen species (ROS) assay, and lactate and dehydrogenase (LDH) release assay. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) expressions were also analyzed by western blotting. Focal ischemia rodent model was used as the in vivo model, and different concentrations of $PineXol^{(R)}$ (1, 10, and 100 mg/kg) were administered. One week after administration, reduction of infarct volume was analyzed by TTC staining. Cell viability of $H_2O_2$-treated SK-N-MC cells significantly increased by pre-treatment of $PineXol^{(R)}$ (p<0.05). $PineXol^{(R)}$ pre-treatment also induced significant decrease of ROS and LDH expressions. However, $PineXol^{(R)}$ did not affect the infarct volume. These results suggest that $PineXol^{(R)}$ has significant neuroprotective effect in vitro, but statistical significance was not confirmed in the in vivo focal ischemia model.

Local Application of NK1 Receptor Antagonists and Pulpal Blood Flow in Cat

  • Kim, Young-Kyung;Chu, Wan-Sik;Lee, Ho-Jeong;Ahn, Dong-Kuk;Yoo, Hyun-Mi;Kim, Sung-Kyo
    • Restorative Dentistry and Endodontics
    • /
    • v.29 no.3
    • /
    • pp.239-248
    • /
    • 2004
  • The purpose of this study was to investigate the influence of NK1 receptor antagonists on the pulpal blood flow (PBF) when applied iontophoretically through the dentinal cavity of the teeth in order to understand whether iontophoretically applied NK1 receptor antagonists can control the pulpal inflammation. Eleven cats were anesthetized with alpha-chloralose and urethane, and substance P (SP) was administered to the dental pulp through the catheterized lingual artery in doses that caused PBF change without the influence of systemic blood pressure. NK1 receptor antagonists were applied iontophoretically to the prepared dentinal cavity of ipsilateral canine teeth of the drug administration, and PBF was monitored. Data were analyzed statistically with paired t-test. PBF increase after iontophoretic application of the NK1 receptor antagonists followed by the intra-arterial administration of SP was significantly less than PBF increase after iontophoretic application of the 0.9% saline followed by the intra-arterial administration of SP as a control (p < 0.05). Iontophoretic application of the NK1 receptor antagonists (0.2~3.4 mM) following the intra-arterial administration of SP resulted in less increase of PBF than the iontophoretic application of the 0.9% saline following the intra-arterial administration of SP as a control (p < 0.05). Therefore. the results of the present study provide evidences that the iontophoretic application is an effective method to deliver drugs to the dental pulp. and that iontophoretically applied NK1 receptor antagonists block SP-induced vasodilation effectively. The above results show the possibility that the iontophoretical application of NK1 receptor antagonists can control the neurogenic inflammation in the dental pulp.

Effect of FC-GT Supplement on Body Fat and Lipid Metabolism in Rats

  • Woo, Myoung-Nam;Jeon, Seon-Min;Kim, Hye-Jin;Yeo, Ji-Young;Ha, Tae-Youl;Choi, Myung-Sook
    • Nutritional Sciences
    • /
    • v.9 no.4
    • /
    • pp.248-258
    • /
    • 2006
  • The objective of this study was to investigate the effects of supplementation of an antiobese functional formula (FC-GT) on body weight and lipid metabolism in rats fed a high-fat diet. Three groups of male Sprague-Dawley rats were fed different diets for 6 weeks: normal control (NC), high-fat (HF), and high-fat supplemented with powdered antiobese functional formula (FC-GT) (5% wt/wt) groups. Although body weight was not significantly different among the groups, relative weights of epididymal and perirenal white adipose tissues were significantly lower in the FC-GT group than in the HF group. FC-GT supplementation significantly lowered the plasma total cholesterol and triglyceride concentrations, whereas it elevated the ratio of HDL-C/total-C and improved the atherogenic index. Hepatic cholesterol and triglyceride concentrations were significantly lowered in the FC-GT group compared to the HF group. The accumulation of hepatic lipid droplets and the epididymal white adipocyte size of the FC-GT group were diminished compared to the HF group. Hepatic HMG-CoA reductase activity was significantly lower in the FC-GT group than in the HF group. Plasma GPT activity was significantly lowered in the FC-GT group compared to the HF group. Additionally, fecal weight was significantly increased in the FC-GT group than in the HF group. In addition, contents of fecal triglyceride and cholesterol were significantly higher in the FC-GT group compared to the other groups. The antioxidant activities of hepatic SOD, CAT, and GR were significantly increased in the FC-GT group compared to the HF group. Hepatic mARS and plasma mARS levels were significantly lowered in the FC-GT group compared to the NC group. Accordingly, we conclude that supplementation of FC-GT improves plasma and hepatic lipid levels in high-fat fed rats.

Single-stranded DNA Enhances the Rate of Product Release During Nucleotide Hydrolysis Reaction by T7 DNA Helicase

  • Kim, Dong-Eun;Jeong, Yong-Joo
    • Bulletin of the Korean Chemical Society
    • /
    • v.27 no.10
    • /
    • pp.1618-1622
    • /
    • 2006
  • Bacteriophage T7 gp4A' is a ring-shaped hexameric DNA helicase that catalyzes duplex DNA unwinding using dTTP hydrolysis as an energy source. To investigate the effect of single-stranded DNA (ssDNA) on the kinetic pathway of dTTP hydrolysis by the T7 DNA helicase complexed with ssDNA, we have first determined optimal concentration of long circular M13 single-stranded DNA and pre-incubation time in the absence of $Mg^{2+}$ which is necessary for the helicase-ssDNA complex formation. Steady state dTTP hydrolysis in the absence of $Mg^{2+}$ by the helicase-ssDNA complex provided $k_{cat}$ of $8.5\;{\times}\;10^{-3}\;sec^{-1}$. Pre-steady state kinetics of the dTTP hydrolysis by the pre-assembled hexameric helicase was monitored by using the rapid chemical quench-flow technique both in the presence and absence of M13 ssDNA. Pre-steady state dTTP hydrolysis showed distinct burst kinetics in both cases, indicating that product release step is slower than dTTP hydrolysis step. Pre-steady state burst rates were similar both in the presence and absence of ssDNA, while steady state dTTP hydrolysis rate in the presence of ssDNA was much faster than in the absence of ssDNA. These results suggest that single-stranded DNA stimulates dTTP hydrolysis reaction by T7 helicase by enhancing the rate of product release step.

The Electrophysiological Characteristics of Medullospinal Tract Cells in Cat Ventrolateral Medulla

  • Lee, Woo-Yong;Kim, Sang-Jung;Kim, Jun
    • The Korean Journal of Physiology
    • /
    • v.25 no.2
    • /
    • pp.211-221
    • /
    • 1991
  • Although the existence of nerve cells which determine the activity of sympathetic nervous system in ventrolateral medulla is advocated recently, there are wide varieties on the location and function of them according to authors. Present study aimed to identify and characterize the medullospinal tract cells in rostral and caudal medulla of cats .which branch to the lateral horn of the upper thoracic spinal cord. Cats were anesthetized with ${\alpha}-chloralose$. The upper thoracic spinal cord and floor of the IVth ventricle were exposed. Medullospinal tract cells in rostral and caudal medulla were identified by anti-dromic stimulation of the intermediolateral nucleus in the upper thoracic cord and then the location and physiological characteristics of these cells were studied. A total of seventy cells in medulla had constant latency and responded to high frequency stimulation to thoracic cord. Among them fifty-six cells were identified as medullospinal tract cells either by collision with spontaneous activities or activities evoked by sciatic nerve stimulation(27/56), or by determining the refractory period (29/56). Thirty-one of these cells branched to the contralateral thoracic spinal cord, twenty-one cells to the ipsilateral side and remaining four cells branched to both sides. The conduction velocity of cells branching to the contralateral side was $29{\pm}2.9\;m/sec$ and that of cells to the ipsilateral side was $39.1{\pm}6.0\;m/sec$. When medulla was devided into two by a horizontal plane at 3 mm rostral to the obex, fifty-one among seventy cells were in the rostral medulla and nineteen were in the caudal medulla. The conduction velocities of these two groups were $21.6{\pm}1.0\;and\;33.3{\pm}3.9\;m/sec$, respectively. In this study, we confirmed the existence of two groups of medullospinal tract cells in rostral and caudal ventrolateral medulla, which branch to the lateral horn of thoracic cord and these cells have relatively few spontaneous activities and rapid conduction velocity, so we concluded that these cells are different from the previously known sympatho-related cells in ventrolateral medulla.

  • PDF

Changes in Antioxidant Enzyme Activity and Physiological Responses to Cadmium and Tributyltin Exposure in the Ark Shell, Scapharca Broughtonii

  • An, Myung-In;An, Kwang-Wook;Choi, Cheol-Young
    • Molecular & Cellular Toxicology
    • /
    • v.5 no.4
    • /
    • pp.273-282
    • /
    • 2009
  • Cadmium (Cd) and tributyltin (TBT) are common contaminants of marine and freshwater ecosystems, and can induce the formation of reactive oxygen species (ROS). These ROS can, in turn, cause oxidative stress. In the present study, we investigated time-related effects of Cd (0.05 and 0.1 ppm) and TBT (5 and 10 ppb) treatment on antioxidant enzyme activity, i.e., the activity of superoxide dismutase (SOD) and catalase (CAT) in the gills and digestive glands of the ark shell, Scapharca broughtonii. In addition, hydrogen peroxide ($H_2O_2$) concentrations, lysozyme activity, and glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) levels were measured in the hemolymph. We found that Cd and TBT treatment significantly increased antioxidant enzyme mRNA expression and activity in the digestive glands and gills in a time-dependent manner. In response to the Cd and TBT treatments, antioxidant enzymes mRNA expression and activity increased up to day 5 in the digestive glands and then decreased by day 7. In the gills, antioxidant enzymes mRNA expression and activity increased up to day 3 and then decreased by day 5. Likewise, $H_2O_2$ concentrations significantly increased up to day 5 and then decreased by day 7. Finally, lysozyme activity decreased during the experimental period, whereas GOT and GPT levels were significantly increased in a time-dependent manner. These results suggest that antioxidant enzymes play an important role in decreasing ROS levels and oxidative stress in ark shells exposed to Cd and TBT.

Molecular Cloning and mRNA Expression of the Hanwoo CAT/enhancer-binding Protein α(C/EBPα) Gene (한우 CCAAT/enhancer-binding protein α(C/EBPα) 유전자의 동정과 mRNA의 발현)

  • Jeoung, Y.H.;Lee, S.M.;Park, H.Y.;Yoon, D.H.;Moon, S.J.;Chung, E.R.;Kang, M.J.
    • Journal of Animal Science and Technology
    • /
    • v.46 no.6
    • /
    • pp.909-916
    • /
    • 2004
  • CCAAT/enhancer binding proteins(C/EBP) are a group of transcription factors expressed during preadipocyte differentiation. In the C/EBPs, C/EBPa plays an important role in lipid deposition and adipocyte differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo CIEBP$\alpha$ The Hanwoo C/EBP$\alpha$DNA includes a 1059 bp open reading frame encoding a protein of 353 amino acids. The CIEBPa amino acid sequences of the Hanwoo show strong conservation with the corresponding sequences reported in other species. The distribution of C/EBP$\alpha$ mRNA in various tissues of Hanwoo aged 12 months were investigated using Northern blotting analysis. The highest expression was detected in adipose tissue and more lower expression was detected in colon and lung. We also identified expression of C/EBPa mRNA in Hanwoo sirloin and adipose tissue aged 12, 26, and 30 months by real-time RT-PCR. The higest expression were detected at 26 months in the sirloin and at 12 and 26 months in the adipose tissue.