• Title/Summary/Keyword: H-uptake

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Characteristics of $[^3H]$-Choline Uptake into Synaptosomes from Rat Hippocampus

  • Lee, Se-Eun;Hong, Seong-Gil;Choe, Rim-Sun
    • Animal cells and systems
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    • v.2 no.2
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    • pp.233-238
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    • 1998
  • Certain basic characteristics of choline uptake in nerve terminals were studied with synaptosomes from rat hippocampus. Synaptosomal $[^3H]$-choline uptake was clarified as specific and high affinity by low Km value(2.2 uM), Na+-dependency and high sensitivity to hemicholinium-3, a competitive inhibitor of choline uptake. Choline uptake into synaptosomes was linearlys related to Na+ concentration and membrane potential. Extracellular Ca2+ modulated the choline uptake, but probably not through increase of intracellular $Ca^{2+}$, because this modulation was not affected the by high $K^+$-depolarization. EGTA (2mM) added for $Ca^{2+}$-free condition had a peculiar effect of decreasing choline uptake. These results suggest that Ca2+ may play an important role in regulating the metabolism of acetylcholine in the nerve terminals directly through the increase of acetylcholine release.

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$[^3H]$ Ouabain Binding and Effect of Ouabain on $^{45}Ca^{2+}$-Uptake in Rat Cardiac Myocytes (쥐 심근 세포의 $[^3H]$ Ouabain 결합과 $^{45}Ca^{2+}}$섭취에 미치는 Ouabain의 영향)

  • 이신웅;김영희;진갑덕
    • YAKHAK HOEJI
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    • v.28 no.3
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    • pp.129-138
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    • 1984
  • Specific [$^{3}H$] ouabain binding and $Ca^{2+}$ -uptake were measured to elucidate the role of high affinity [$^{3}H$] ouabain binding site in rat cardiac myocytes which contain 65% of rod cells. High affinity [$^{3}$H] ouabain binding site, which is about 3% of total pump sites, with apparent dissociation constant ($K_{D}$) of $1.1{\times}10^{-7}M$ and maximum binding site concentration (Bmax) of 1.2 pmol/mg protein ($1.754{\times}10^{5}cells$) were identified. At the concentration of $10^{-7}M$ to $10^{-4}M$, ouabain produced concentration dependent increase in $Ca^{2+}$-uptake of myocytes. The effect of ouabain on $Ca^{2+}$-uptake was not effected by membrane depolarization (elevated K+ in incubation medium) or verapamil. These results suggest that in rat ventricular myocytes the ouabain receptor complex to high affinity site may increase Na+ - $Ca^{2+}$ exchange across the sarcolemmal membrane by inhibition of Na+, K+ - ATPase.

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Transport Properties of Aromatic Amino Acids by Amino Acid Transporter TAT1 (아미노산 수송체 TAT1에 의한 방향족 아미노산의 수송특성)

  • 김윤배;김명수;윤정훈;박주철;국중기;정해만;최봉규;정규용;김종근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.775-781
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    • 2002
  • The T-type amino acid transporter 1 (TATI) is a Na$^{+}$-independent amino acid transporter which selectively trans- ports aromatic amino acids subserving the amino acid transport system T. To understand the transport properties of aromatic amino acids by human TAT1 (hTATl ), we have examined the hTATl -mediated aromatic amino acid transports using a Xenopus laeuis oocyte expression system. When expressed in Xenopin laeuis oocytes, hTATl induced L- [$^{14}$ C]tryptophan transport which was not dependent on Na$^{+}$ or Cl$^{[-10]}$ in the medium. Uptake was time-dependent and exhibited a linear dependence on incubation time up to 30 min. The L- ($^{14}$ C)tryptophan uptake was highly inhibited by L-isomers of tryptophan, tyrosine and phenylalanine, whereas other L-amino acids did not inhibit hTATl -mediated L- ($^{14}$ C)tryptophan uptake. The hTATl induced the relatively low-affinity transport of aromatic amino acids such as L- ($^{14}$ C)tryptophan, L- ($^{14}$ C)tyrosine and L- ($^{14}$ C)phenylalanine (Km values: 450~750 $\mu$M), consistent with the properties of classical amino acid transport system T. The L- ($^{14}$ C)tryptophan uptake did not show any remarkable pH dependence within the pH range of 5.5 to 8.5. The time-dependent efflux of L- ($^{14}$ C)tryptophan was detected from the oocytes expressing hTATl, which was not affected by the presence or absence of L-tryptophan in the extracellular medium, indicating that hTATl-mediated transport is due to the facilitated diffusion. Expression of hTATl in Xenopu laevis oocytes induced the transport of tryptophan, tyrosine and phenylalanine, indicating that hTATl is a transporter subserving system T These results suggest that hTATl has essential roles in the absorption of aromatic amino acids from epithelial cells to the blood stream. Hecause hTATl is proposed to be crucial to the efficient absorption of aromatic amino acids from intestine and kidney, its defect such as blue diaper syndrome could be involved in the disruption of aromatic amino acid transport.ort.

Increased Uptake of Cadmium by Surfactants in a Cadmium-Tolerant Yeast (카드뮴 내성효모의 카드뮴축적에 미치는 계면활성제의 영향)

  • 송형의
    • Journal of Environmental Health Sciences
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    • v.22 no.2
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    • pp.104-113
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    • 1996
  • Cadmium uptake by growing and nongrowing (intact) cells of a chdmium-tolerant yeast Hansenula anomala B-7 in the presence of surfactants was studied. In growing cultures the addition of Triton X-100 or Tween 80 increased cadmium uptake by about 30% with no inhibition of cell growth, and in intact cells Triton X-100 increased cadmium accumulation by about 80% compared to surfactant-free controls. Considering balance between increased uptake and pollution, the addition of 0.1% Triton X-100 was preferable. By the mixed addition with defoamer silicone, during growth of cells Tween 80 or Triton X-100 enhanced uptake efficiency of cadmium compared to its single addition, whereas in intact biomass each of surfactants tested had no significant effect on cadmium uptake. The uptake of cadmium was observed to rise sharply to a maximum and then declined with increasing pH, and maximum accumulation of cadmium by growing and intact cells occurred at the pH of 6.0 and 7.0, respectively. A significant increase in cadmium uptake occurred with shaking culture. Cadmium uptake by growing and intact cells was almost completed during the culture time of 72 or 24 hrs, respectively. Scalded cells sorbed much more cadmium-ion than living cells.

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Variations in Nutrients and CO2 Uptake with Growth of Undaria pinnatifida from the South Coast of Korea (미역 (Undaria pinnatifida)의 생장에 따른 영양염과 CO2 흡수율 변화)

  • Shim, Jeong-Hee;Hwang, Jae-Ran;Lee, Jae-Seong;Kim, Jong-Hyun;Kim, Sung-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.6
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    • pp.679-686
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    • 2010
  • To investigate the contribution of macroalgae to biogeochemical nutrients and carbon cycles, we measured the uptake rates of nutrients and $CO_2$ by Undaria pinnatifida using an incubation method in an acrylic chamber. From January to March 2010, U. pinnatifida was sampled at Ilkwang, a well-known area of macroalgae culture in Korea. The initial and final concentrations of nutrients, dissolved oxygen, total alkalinity, and pH of the chamber water were measured, and production/uptake rates were calculated using concentration changes, chamber volume, and incubation time. The production rate of dissolved oxygen by U. pinnatifida (n = 32) was about $5.4{\pm}4.0\;{\mu}mol\;g_{fw}^{-1\;}h^{-1}$. The uptake rate of total dissolved inorganic carbon (TDIC), calculated by total alkalinity and pH, was $7.9{\pm}6.5\;{\mu}mol\;g_{fw}^{-1}\;h^{-1}$. Nutrients uptake averaged $141.7{\pm}119.2$ nmol N $g_{fw}^{-1}\;h^{-1}$ and $15.0{\pm}9.1$ nmol P $g_{fw}^{-1}\;h^{-1}$. A positive linear correlation ($r^2$ = 9.6) existed between the production rate of dissolved oxygen and the uptake rate of total dissolved inorganic carbon, suggesting that these two factors serve as good indicators of U. pinnatifida photosynthesis. The relationships between fresh weight and uptake rates of nutrients and $CO_2$ suggested that younger specimens (<~50 g fresh weight) are much more efficient at nutrients and $CO_2$ uptake than are specimens >50 g. The amount of carbon uptake by the total biomass of U. pinnatifida in Korea during the year of 2008 was about 0.001-0.002% of global ocean carbon uptake. Thus, more research should be focused on macroalgae-based biogeochemical cycles to evaluate the roles and contributions of macroalgae to the global carbon cycle.

Antidiabetic Activities of Extract from Malva verticillata Seed via the Activation of AMP-Activated Protein Kinase

  • Jeong, Yong-Tae;Song, Chi-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.921-929
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    • 2011
  • Stimulation of AMP-activated protein kinase (AMPK) signaling followed by increase of glucose uptake in L6 myotubes were studied with organic solvent extract of Malva verticillata (MV) seeds. Ethanol extract of M. verticillata seeds (MVE) significantly increased the phosphorylation level of AMPK, acetyl-CoA carboxylase (ACC), and glucose uptake in L6 myotube cells. The MVE was fractionated with n-hexane (MVE-H), chloroform (MVE-C), ethylacetate (MVE-E), n-butanol (MVE-B), and water (MVE-W). MVE-H (150 ${\mu}g$/ml) showed the highest phosphorylating activity and increased glucose uptake by 2.3-fold. Oral administration of MVE-H (40 mg/kg) for 4 weeks to type 2 diabetic (db/db) mice reduced non-fasting and fasting blood glucose levels by 17.1% and 23.3%, respectively. Phosphorylation levels of AMPK and ACC in the soleus muscle and liver tissue of db/db mice were significantly increased by the administration of MVE-H. MVE-H was further fractionated using preparative HPLC to identify the AMPK-activating compounds. The NMR and GC-MS analyses revealed that ${\beta}$-sitosterol was a major effective compound in MVE-H. Phosphorylation levels of AMPK and ACC, and glucose uptake were significantly increased by the treatment of MVE-S (${\beta}$-sitosterol) isolated from M. verticillata to L6 cells, and these effects were attenuated by an AMPK inhibitor (Compound C) pretreatment. These results, taken together, demonstrate that increased glucose uptake in L6 myotubes by MVE-H treatment is mainly accomplished through the activation of AMPK. Our finding suggests that the extract isolated from M. verticillata seed would be beneficial for the treatment of metabolic disease including type 2 diabetes and hyperlipidemia.

The Change of Taurine Transport in Variable Stress States through the Inner Blood-Retinal Barrier using In Vitro Model

  • Kang, Young-Sook;Lee, Na-Young;Chung, Yeon-Yee
    • Biomolecules & Therapeutics
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    • v.17 no.2
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    • pp.175-180
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    • 2009
  • Taurine is the most abundant free amino acid in the retina and transported into retina via taurine transporter (TauT) at the inner blood-retinal barrier (iBRB). In the present study, we investigated whether the taurine transport at the iBRB is regulated by oxidative stress or disease-like state in a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB) used as an in vitro model of iBRB. First, [$^3H$]taurine uptake and efflux by TR-iBRB were regulated in the presence of extracellular $Ca^{2+}$. [$^3H$]Taurine uptake was inhibited and efflux was enhanced under $Ca^{2+}$ free condition in the cells. In addition, oxidative stress inducing agents such as tumor necrosis factor-$\alpha$ (TNF-$\alpha$), lipopolysaccharide (LPS), diethyl maleate (DEM) and glutamate increased [$^3H$]taurine uptake and decreased [$^3H$]taurine efflux in TR-iBRB cells. Whereas, 3-morpholinosydnonimine (SIN-1), which is known to NO donor decreased [$^3H$]taurine uptake. Lastly, TR-iBRB cells exposed to high glucose (25 mM) medium and the [$^3H$]taurine uptake was reduced about 20% at the condition. Also, [$^3H$]taurine uptake was decreased by cytochalasin B, which is known to glucose transport inhibitor. In conclusion, taurine transport in TR-iBRB cells is regulated diversely at extracellular $Ca^{2+}$, oxidative stress and hyperglycemic condition. It suggested that taurine would play a role as a retinal protector in diverse disease states.

Effect of pH in Irrigation Water on the Growth, Yield, and Grain Quality of Rice (관개용수 pH가 벼 생육, 수량, 미질에 미치는 영향(I))

  • Choi, Sun-Hwa;Kim, Ho-Il;An, Yeul;Huh, Yoo-Man
    • Proceedings of the Korean Society of Agricultural Engineers Conference
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    • 2003.10a
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    • pp.551-554
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    • 2003
  • This study was carried out to investigate the effects of the pH of irrigation water on the growth, yield, and grain quality of rice. It acquire fundamental knowledges to set up irrigation water quality standards. The pot experiment was conducted with 5 treatments using irrigation waters with various pH values(control, 4, 6, 8, 10) and replicated four times with randomized block design. The results of this study showed that the uptake of N, P, and K, Ripened grain ratio and yield of rice tended to be reduced at the irrigation water of pH 4 and pH 10. P uptake, Ripened grain ratio and yield of rice at pH 4 water were significantly lower than the control. K uptake at pH 10 water was significantly lower than the control. Plant height, SPAD value and protein content of rice were not affected by the pH of irrigation water.

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UPTAKE OF α-AMINOISOBUTYRIC ACID (AlB) BY ROOSTER SPERMATOZOA

  • Fujihara, N.;Koga, O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.3 no.2
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    • pp.91-96
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    • 1990
  • This experiment was designed to determine whether ${\alpha}$-aminoisobutyric acid (AIB) can be used to predict membrane function of spermatozoa by measuring the uptake of AIB by fresh, stored and frozen-thawed rooster spermatozoa. When spermatozoa were stored at low temperature ($0{\sim}3^{\circ}C$) for 24 h. no difference was found in AIB uptake compared with fresh spermatozoa, whereas storage for 48 h resulted in a slight increase in AIB uptake by spermatozoa. On the one hand, the uptake of AIB by frozen-thawed spermatozoa was less than that by fresh spermatozoa. This suggests possibility of a different membrane transport system between spermatozoa preserved at low temperature ($0{\sim}3^{\circ}C$) and those frozen-thawed. Glycerol used as cryoprotectant may modify rooster sperm membrane in a different manner from cold preservation. Ouabaine ($10^{-4}M$) caused a slight decrease in AIB uptake, but caffeine ($10^{-2}M$) did not influence spermatozoal AIB uptake. These results indicate a successful application of AIB to rooster spermatozoa as a mean for measuring sperm membrane function and suggest a possible alteration of membrane transport system in rooster spermatozoa between cold ($0{\sim}3^{\circ}C$) and cryopreservation ($-196^{\circ}C$).

Inhibition of Sarcoplasmic Reticulum $Ca^{2+}$ Uptake by Pyruvate and Fatty Acid in H9c2 Cardiomyocytes: Implications for Diabetic Cardiomyopathy

  • Lee, Eun-Hee;Lee, Hye-Kyung;Kim, Hae-Won;Kim, Young-Hoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.4
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    • pp.195-201
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    • 2005
  • High extracellular glucose concentration was reported to suppress intracellular $Ca^{2+}$ clearing through altered sarcoplasmic reticulum (SR) function. In the present study, we attempted to elucidate the effects of pyruvate and fatty acid on SR function and reveal the mechanistic link with glucose-induced SR dysfunction. For this purpose, SR $Ca^{2+}$-uptake rate was measured in digitonin-permeabilized H9c2 cardiomyocytes cultured in various conditions. Exposure of these cells to 5 mM pyruvate for 2 days induced a significant suppression of SR $Ca^{2+}$-uptake, which was comparable to the effects of high glucose. These effects were accompanied with decreased glucose utilization. However, pyruvate could not further suppress SR $Ca^{2+}$-uptake in cells cultured in high glucose condition. Enhanced entry of pyruvate into mitochondria by dichloroacetate, an activator of pyruvate dehydrogenase complex, also induced suppression of SR $Ca^{2+}$-uptake, indicating that mitochondrial uptake of pyruvate is required in the SR dysfunction induced by pyruvate or glucose. On the other hand, augmentation of fatty acid supply by adding 0.2 to 0.8 mM oleic acid resulted in a dose-dependent suppression of SR $Ca^{2+}$-uptake. However, these effects were attenuated in high glucose-cultured cells, with no significant changes by oleic acid concentrations lower than 0.4 mM. These results demonstrate that (1) increased pyruvate oxidation is the key mechanism in the SR dysfunction observed in high glucose-cultured cardiomyocytes; (2) exogenous fatty acid also suppresses SR $Ca^{2+}$-uptake, presumably through a mechanism shared by glucose.