• Title, Summary, Keyword: Acetate Transport

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Production of Leptin in E. coli and Its Effect on Glucose and Acetate Transport and Expression of Uncoupling Protein-2 Gene in Adipose Tissues of Korean Cattle (Hanwoo)

  • Kim, K.S.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1062-1068
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    • 2004
  • Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

Retardation of Drug Transport through Pig Ear Skin by Liposome-Hydrogel

  • Bae, Soo-Kyoung;Kim, Jin-Chul;Kim, Jong-Duk
    • Proceedings of the Korean Biophysical Society Conference
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    • pp.21-21
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    • 1997
  • Transport of drug entrapped in a liposome-hydrogel formulation was significantly retarded in an in vitro topical application. Liposomes containing hydrocortisone acetate, a hydrophobic antiinflammatory agent, were prepared by the precipitation method, and the liposomal suspension was mixed with hydrogel into a semisolid gel-type ointment.(omitted)

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Effects of Lead on the Ultrastructure ana the Electron Transport System of Mitochondria of Mouse Kidney (납(Pb)이 생쥐 신장세포에 미토콘드리아 미세구조 및 전자전달계에 미치는 영향)

  • Lim, Seung-Sub;Yoo, Chang-Kyu;Choe, Rim-Soon
    • Applied Microscopy
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    • v.17 no.2
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    • pp.55-71
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    • 1987
  • To investigate the effects of lead on the electron transport system and ultrastructure of mouse kidney mitochondria, various lead acetate concentrations were treated in vitro and respiration rate, enzyme activities were measured. Ultrastructural changes at state IV respiration were also observed. To compare with in vivo experiments, mouse were injected intraperitoneally of 100 mg lead acetate per kg body weight and state IV respiration rate and enzyme activities were measured. Ultrastructure of renal proximal tubular cells were also observed. In in vitro treatement, decreased state IV respiration, decreased enzyme activities, ruptured membranes and inhibition of condensed to orthodox transformation were observed. In in vivo treatment, decreased state IV respiration and decreased enzyme activities were observed after 24 hrs of i.p. injection. Cytochrome c oxidase activity showed twice the inhibition compared to NADH-CoQ reductase activity at 24 hrs. Continuous decreased state IV respiration was observed after 48 and 72 hrs of injection, however, the enzyme activities were increased to control level. Lead-protein complex which probably inhibits the toxic effects of lead appeared. To conclude, dominant effect of lead on the electron transport system appeared at cytochrome c oxidase activity, and the increased enzyme activities may be a result of appearance of lead-protein complex.

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Gas Separation of Pyrolyzed Polymeric Membranes: Effect of Polymer Precursor and Pyrolysis Conditions

  • Jung, Chul-Ho;Kim, Gun-Wook;Han, Sang-Hoon;Lee, Young-Moo
    • Macromolecular research
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    • v.15 no.6
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    • pp.565-574
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    • 2007
  • In this study, five representative, commercially available polymers, Ultem 1000 polyetherimide, Kapton polyimide, phenolic resin, polyacrylonitrile and cellulose acetate, were used to prepare pyrolyzed polymer membranes coated on a porous {\alpha}-alumina$ tube via inert pyrolysis for gas separation. Pyrolysis conditions (i.e., final temperature and thermal dwell time) of each polymer were determined using a thermogravimetric method coupled with real-time mass spectroscopy. The surface area and pore size distribution of the pyrolyzed materials derived from the polymers were estimated from the nitrogen adsorption/desorption isotherms. Pyrolyzed membranes from polymer precursors exhibited type I sorption behavior except cellulose acetate (type IV). The gas permeation of the carbon/{\alpha}-alumina$ tubular membranes was characterized using four gases: helium, carbon dioxide, oxygen and nitrogen. The polyetherimide, polyimide, and phenolic resin pyrolyzed polymer membranes showed typical molecular sieving gas permeation behavior, while membranes from polyacrylonitrile and cellulose acetate exhibited intermediate behavior between Knudsen diffusion and molecular sieving. Pyrolyzed membranes with molecular sieving behavior (e.g., polyetherimide, polyimide, and phenolic resin) had a $CO_2/N_2$ selectivity of greater than 15; however, the membranes from polyacrylonitrile and cellulose acetate with intermediate gas transport behavior had a selectivity slightly greater than unity due to their large pore size.

The Transport of a Hepatoprotective Agent, Isopropryl 2-(1-3-dithiethane-2-ylidene)-2[N-(4-methyl-thiautole-2-yl) carbamoyl] Acetate (YH439), across Caco-2 Cell Monolayers

  • Park, Hyeon-Woo;Chung, Suk-Jae;Lee, Myung-Gull;Shim, Chang-Koo
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.584-589
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    • 2001
  • Isopropryl 2-(1-3-dithiethane-2-ylidene)-2 [N-(4-methyl-thiazole-2-yl) carbamoyl] acetate (YH439) is currently under phase ll clinical trials by the Yuhan Research Center for use as a hepatoprotective agent. Unfortunately, the oral bioavailbility of YH439, which is sparingly soluble in water (i.e., $0.3{\;}\mu\textrm{g}/ml{\;}or{\;}0.91{$\mu}M$ at room temperature), reportedly, is negligibleregardless of the dose administered to rats in the 10-300 mg/kg range. The bioavailability of the compound increased up to 24%, when administered in the form of a micellar solution ($700{\;}\mu\textrm{g}/ml$or 2.1 mM for YH439) at a dose of 10 mg/kg, suggesting that its limited solubility is associated with its negligible bioavailability. In order to obtain additional informmation concerning the bioavailability of YH439, the mechanism(s) involved in gastrointestinal (Gl) absorption were investigated in the present study. For this purpose, the transport of YH430 across a Caco-2 cell monolayer was measured in a $Transwell^{\circledR}$. A permeability of $4.07{\times}10^{-5}{\;}cm/s$ was obtained for the absorptive (i.e., apical to basolateral direction) transport of $0.42{\mu}M$ YH439, implicating that the in vivo Cl absorption is nearly complete. The absorptive transport exhibited a slight concentration-dependency with an intrinsic clearance ($CL_{i}$) of $0.38{\mu}L/{\textrm{cm}^2}/sec$, which accounted for 28.1% of the total intrinsic clearance (i.e., $CL_i$ plus the intrinsic clearance for the linear component) of the transport. Thus, saturation of the absorption process appears to be a minor factor in limiting the bioavailability of the compound. The apparent permeability of YH439 from the basolateral to the apical direction (i.e., efflux, $6.67{\times}10^{-5}{\;}cm/s$) was comparable to that for absorptive transport, but, interestingly, a more distinct concentration-dependency was observed for this transport. However, the efflux does not appear to influence the bioavailability of the compound, as evidenced by the sufficiently high permeability in the absorption direction. Rather, a reportedly extensive first-pass hepatic metabolism appears to be a principal factor in limiting the bioavailability. In this respect, reducing the first-pass metabolism by some means would lead to a higher bioavailability of the compound. Thus, elevation of the absorption rate of YH439 becomes a necessity. From a practical point of view, increasing the concentration of YH439 in the Cl fluid appears to be a feasible way to increase the absorption rate, because the compound is primarily absorbed via a linear mechanism. In summary, the solubilization of YH439, as previously demonstrated for a micellar solution of the compound, appears to be a practical way to increase the oral bioavailability of YH439.

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Studies on the Transport of Organic Acids in the Rabbit Kindey Slice, with Special Reference to the Role of Various Electrolytes (가토신피질절편(家兎腎皮質切片)에서의 유기산이동(有機酸移動)에 관(關)한 연구(硏究) -특(特)히 전해질(電解質)의 영향(影響)에 대(對)하여-)

  • Chung, Soon-Tong
    • The Korean journal of physiology & pharmacology
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    • v.2 no.1
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    • pp.59-71
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    • 1968
  • The uptake of phenolsulfonphthalein (PSP) and of paraaminohippuric acid (PAH) by cortical slices of the rabbit kidney was investigated while varying the composition of medium. The overall uptake of these substances displayed typical active transport characteristics and was significantly enhanced in presence of acetate. When the phosphate buffer was used the optimal pH was 7.4 for both substances. However, when the tris-buffer was used the optimal pH was 7.4 for PSP and 8.3 for PAH. Removal of $Na^+$ from the medium resulted in a significant reduction in the uptake. Similar results, though lesser in magnitude, were obtained when either $K^+\;or\;Ca^{++}$ was removed from the medium. However, there was no additive effect when $K^+\;and/or\;Ca^{++}$ were additionally removed from the $Na^+-free$ medium. The presence of ${NH_4}^+$ greatly reduced while $Li^+\;and\;Mg^{++}$ moderately reduced the uptake of both substances. However, choline had no effect. In substrate-leached slices, acetate greatly enhance the uptake of organic acids; but this action was not demonstrable in absence of $Na^+,\;K^+\;or\;Ca^{++}$.

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Characteristics of p-Aminohippurate (PAH) Transport in Basolateral Membrane Vesicles of Rabbit Kidney Cortex -Effect of Anion- (신장 피질 기저막 소포에서 p-Aminohippurate 이동의 특성에 대한 연구 -음이온의 효과-)

  • Lee, Sang-Ho;Chung, Jin-Sup;Kim, Yong-Keun
    • The Korean journal of physiology & pharmacology
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    • v.20 no.2
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    • pp.225-235
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    • 1986
  • Effects of anions on p-Aminohippurate (PAH) transport across the basolateral membrane (BLM) were studied. Basolateral membrane vesicles were isolated from rabbit renal cortex by using a self-orienting Percoll-gradient centrifugation and $Mg^{2+}-precipitation$. The characteristics of the membrane vesicles was examined by marker enzyme activity, membrane orientation and transport studies. The Na-K-ATPase activity in the fraction containing BLM vesicles was enriched 9·fold, and the alkaline phosphatase activity in the fraction containing BBM vesicles was increased 9-fold, compared with those of the homogenate. The transport properties of the two membrane preparations were studied by a rapid filtration technique. The uptake of PAH by BLM was sensitive to changes in medium osmolarity and inhibited by probenecid. When the uptake of $50{\mu}M$ PAH in voltage-clamped BLM vesicles was determined in the presence of various anions in the incubation medium, cis inhibitions by $SO_4\;and\;SSO_3$ were observed in the presence of sodium gradient (out>in). Sodium-dependent PAH uptake was inhibited competitively by external $SO_4$ PAH uptake in BLM vesicles loaded with 20 mM acetate and $SO_4\;or\;200\;{\mu}M$ PAH was significantly stimulated as compared with unloaded vesicles. The extent of trans-stmulation of PAH uptake by $SO_4$. was increased with the inside concentration of $SO_4$. This trans-stimulatory effect by $SO_4$, was observed to be additive in the presence of Na gradient and completely inhibited by 2 mM probenecid and 1 mM SITS. These results demonstrate that PAH/anion exchange is present in BLM of renal cortex and in this exchange mechanism inorganic as well as organic anions are involved as substrates.

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Transport Properties of Crosslinked Poly Vinyl Alcohol Membrane in Pervaporation

  • Lee, Chul-Haeng;Hong, Won-Hi
    • Proceedings of the Membrane Society of Korea Conference
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    • pp.92-93
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    • 1996
  • PVA membrane was widely used in the dehydration pervaporation process. PVA membrane showed remakable selectivity towed water and an excellent film-forming polymer, with a good resistance to orgamic solvents but it has poor stability in aqueous mixtures. Generally the PVA is manufactured by the hydrolysis reaction from poly vinyl acetate(PVAc) and so the degree of PVA hydrolysis is a major parameter for properties of PVA membrane such as the crystallinity and polarity.

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The Growth Yield of Desulfovibrio desulfuricans M6 on Different Substrates

  • Park, Doo-Hyun;Shin, Chul-Su;Kim, Byung-Hong;Shin, Pyung-Kyun
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.232-237
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    • 1996
  • Growth yield of Desulfovibrio desulfuricans M6 was measured using different substrates. The cell yield of fermentative growth on pyruvate was 6.22 g cell $mol^{-l}$ pyruvate. Since 1 ATP is available from substrate-level phosphorylation from the oxidation of pyruvate to acetate, $Y_{ATP}$ of the bacterium should be the same as $Y_{pyruvate}$ (6.22 g cell $mol^{-l}$ ATP). The cell yields of the bacterium on different electron donors were measured with sulfate as the electron acceptor. Cell yields on lactate, pyruvate and $H_2$ were 9.39, 13.76 and 8.45 g cell $mol^{-l}$ substrate, respectively. From these figures ATP available from electron-transport phosphorylation (ETP) of the electron donors used was calculated. ATP produced by ETP of each electron donnor were 1.71 from pyruvate, 1.51 from lactate and 1.76 from $H_2$. These values show that electrons from the oxidation of lactate to pyruvate are consumed to reduce sulfate through a reverse electron transport mechanism requiring 0.2 ATP for each pair of electrons. Based on these results, discussions are made on the electron transport mechanism in the bacterium.

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