• Title/Summary/Keyword: NCX

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Changes of Cytosolic $Ca^{2+}$ under Metabolic Inhibition in Isolated Rat Ventricular Myocytes

  • Kang, Sung-Hyun;Kim, Na-Ri;Joo, Hyun;Youm, Jae-Boum;Park, Won-Sun;Warda, Mohamed;Kim, Hyung-Kyu;Von Cuong, Dang;Kim, Tae-Ho;Kim, Eui-Yong;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.5
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    • pp.291-298
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    • 2005
  • To characterize cytosolic $Ca^{2+}$ fluctuations under metabolic inhibition, rat ventricular myocytes were exposed to $200{\mu}M$ 2,4-dinitrophenol (DNP), and mitochondrial $Ca^{2+}$, mitochondrial membrane potential (${\Delta}{\Psi}m$), and cytosolic $Ca^{2+}$ were measured, using Rhod-2 AM, TMRE, and Fluo-4 AM fluorescent dyes, respectively, by Laser Scanning Confocal Microscopy (LSCM). Furthermore, the role of sarcolemmal $Na^+$/$Ca^{2+}$ exchange (NCX) in cytosolic $Ca^{2+}$ efflux was studied in KB-R7943 and $Na^+$-free normal Tyrode's solution (143 mM LiCl ). When DNP was applied to cells loaded with Fluo-4 AM, Fluo-4 AM fluorescence intensity initially increased by $70{\pm}10$% within $70{\pm}10$ s, and later by $400{\pm}200$% at $850{\pm}45$ s. Fluorescence intensity of both Rhod-2 AM and TMRE were initially decreased by DNP, coincident with the initial increase of Fluo-4 AM fluorescence intensity. When sarcoplasmic reticulum (SR) $Ca^{2+}$ was depleted by $1{\mu}M thapsigargin plus $10{\mu}M ryanodine, the initial increase of Fluo-4 AM fluorescence intensity was unaffected, however, the subsequent progressive increase was abolished. KB-R7943 delayed both the first and the second phases of cytosolic $Ca^{2+}$ overload, while $Na^+$-free solution accelerated the second. The above results suggest that: 1) the initial rise in cytosolic $Ca^{2+}$ under DNP results from mitochondrial depolarization; 2) the secondary increase is caused by progressive $Ca^{2+}$ release from SR; 3) NCX plays an important role in transient cytosolic $Ca^{2+}$ shifts under metabolic inhibition with DNP.

Effects of Web-Based Nutrition Counseling on Nutrient Intake and Blood Glucose in Type II Diabetic Patients (제 2형 당뇨병 환자의 영양섭취 및 혈당에 웹기반 영양상담이 미치는 영향)

  • Park, Sun-Young;Han, Ji-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.9
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    • pp.1398-1406
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    • 2005
  • The purpose of this study was to investigate effects of web-based nutrition counseling on nutrient intake and blood glucose in type ll diabetic Patients. Forty type II diabetic patients, twenty one of them were diabetic patients without complication (Ncx-DM) and nineteen of them were diabetic patients with complication (Cx-DM), participated in a web-based nutrition counseling program. At the first nutrition counseling, the patients were counselled through interview and then follow up nutrition counseling was accomplished four times during eight weeks through tile web-based internet program. Various markers of disease risk including anthropometric indices, nutrient intake and blood glucose were measured before and after the nutrition counseling. After the nutrition counseling, body mass index and waist circumference decreased in both group but did not change significantly. Fasting blood glucose significantly decreased from 153.9 mg/dL to 139.0 mg/dL (p<0.05) in NCX-DM and from 178.2 mg/dL to 128.5 mg/dL (p<0.01) in Cx-DM after the nutrition counseling. Glycosylated hemoglobin level decreased from $9.3\%$ to $8,7\%$ in Ncx-DM and significantly decreased from $9.7\%$ to $7.8\%$ (p<0.01) in Cx-DM after the nutrition counseling. In addition, total cholesterol and LDL-cholesterol level significantly decreased in both group (P<0,05) after the nutrition counseling. Energy intake decreased significantly in Ncx-DM (P<0.05) and Cx-DM (p<0.01). Although the nutrient intake did not change significantly, the nutrient intake was improved after the nutrition counseling. Therefore, this study shows that the web-based nutrition counseling is effective in improving energy and nutrient intake and influences positively in blood glucose and serum lipids of the patients. In addition, these results indicate that the internet presents us with potential as a new medium for nutrition counseling in informationized society.

$Na^+/Ca^{2+}$ exchanger (NCX)-2, a temporal factor in regulation of synaptic plasticity and cognition

  • Shin, Hee-Sup
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.21-21
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    • 2003
  • The role of a postsynaptic rise of [C $a^{2+}$]$_{i}$ in the induction of LTP and LTD has been well established. Both the levels and the duration of elevated [C $a^{2+}$]$_{i}$ are important in synaptic plasticity. LTP and LTD could be selectively induced according to intracellular $Ca^{2+}$ concentration. Although the specificity of $Ca^{2+}$ signaling can be achieved not only by amplitude but also by the frequency and duration of the calcium transient, the effects of changing amplitudes of $Ca^{2+}$ transients on synaptic plasticity have been extensively documented, but not so the effects of temporal changes.changes..

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Xylanase supplementation in energy-deficient corn-based diets: impact on broiler growth, nutrient digestibility, chyme viscosity and carcass proximates

  • Bernadette Gerpacio Sta. Cruz;Jun Seon Hong;Myunghwan Yu;Elijah Ogola Oketch;Hyeonho Yun;Dinesh D. Jayasena;Jung-Min Heo
    • Animal Bioscience
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    • v.37 no.7
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    • pp.1246-1254
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    • 2024
  • Objective: The goal of the current study was to investigate the impact of various concentrations of xylanase in energy-deficient corn-based diets on the growth performance, carcass characteristics, nutrient digestibility, and digesta viscosity in broilers from 7 to 35 days of age. Methods: A total of 280 seven-day-old Ross 308 broilers were randomly allocated to one of the five dietary treatments following a completely randomized design with 8 replicates and 7 birds per cage. The treatments were: i) positive control (PC, without xylanase); ii) NC-1 (80 kcal/kg ME reduced from PC); iii) NC-2 (100 kcal/kg ME reduced from PC); iv) NCX-1 (NC-1 + 2,000 U/kg xylanase); and v) NCX-2 (NC-2 + 3,000 U/kg xylanase). Body weight, weight gain, feed intake, and feed conversion ratio were determined weekly to evaluate growth performance. One bird per pen was sacrificed for ileal digesta collection to determine the viscosity and digestibility of energy, dry matter, crude protein on days 24 and 35, however breast and leg meat samples were obtained for proximate analysis (moisture, crude protein, fat, and ash) on day 35. Results: Birds fed diets supplemented with xylanase regardless of the amount had higher (p<0.05) body weights, daily gains, and improved feed efficiency compared to NC diets all throughout the experimental period. Feed intake was not affected (p>0.05) by the addition of xylanase. Moreover, lowered (p<0.05) viscosity of the ileal digesta were observed upon xylanase inclusion in the diets compared to the birds fed NC diets on day 24. Ileal nutrient digestibility and meat proximate composition were not affected (p>0.05) by xylanase. Conclusion: The present study indicated that the xylanase at 2,000 U/kg and 3,000 U/kg levels compensates for the 80 kcal/kg and 100 kcal/kg dietary energy levels, respectively, without having adverse effects on the growth performance, carcass characteristics, nutrient digestibility, and digesta viscosity of broilers.

Calumenin Interacts with SERCA2 in Rat Cardiac Sarcoplasmic Reticulum

  • Sahoo, Sanjaya Kumar;Kim, Do Han
    • Molecules and Cells
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    • v.26 no.3
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    • pp.265-269
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    • 2008
  • Calumenin, a multiple EF-hand $Ca^{2+}$ binding protein is located in the SR of mammalian heart, but the functional role of the protein in the heart is unknown. In the present study, an adenovirus gene transfer system was employed for neonatal rat heart to examine the effects of calumenin over-expression (Calu-OE) on $Ca^{2+}$ transients. Calu-OE (8 folds) did not alter the expression levels of DHPR, RyR2, NCX, SERCA2, CSQ and PLN. However, Calu-OE affected several parameters of $Ca^{2+}$ transients. Among them, prolongation of time to 50% baseline ($T_{50}$) was the most outstanding change in electrically-evoked $Ca^{2+}$ transients. The higher $T_{50}$ was due to an inhibition of SERCA2-mediated $Ca^{2+}$ uptake into SR, as tested by oxalate-supported $Ca^{2+}$ uptake. Furthermore, co-IP study showed a direct interaction between calumenin and SERCA2. Taken together, calumenin in the cardiac SR may play an important role in the regulation of $Ca^{2+}$ uptake during the EC coupling process.

Cardiac hypertrophy and abnormal $Ca^{2+}$ handling in transgenic mice overexpressing jnnctate

  • Hong, Chang-Soo;Cho, Myeong-Chan;Kwak, Yong-Geun;Chane, Soo-Wan;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.52-52
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    • 2003
  • Junctate is a newly identified integral ER/SR membrane $Ca^{2+}$ binding protein, which is an alternative splicing form of the same gene generating aspartyl $\square$-hydroxylase and junctin. To elucidate the functional role of junctate in heart, transgenic (TG) mice overexpressing mouse cardiac junctate-1 under the control of mouse $\square$$^{~}$ myosin heavy chain promoter were generated. Overexpression of junctate in mouse heart resulted in cardiac hypertrophy, increased fibrosis, bradycardia, arrhythmias and impaired contractility. Overexpression of junctate also led to down-regulation of SERCA2, calsequestrin, calreticulin and RyR, but to up-regulation of NCX and PMCA. The SR $Ca^{2+}$ content decreased and the L-type $Ca^{2+}$ current density and the action potential durations increased in TG cardiomyocytes, which could be the cause for the bradycardia in TG heart. The present work has provided an important example of pathogenesis leading to cardiac hypertrophy and arrhythmia, which was caused by impaired $Ca^{2+}$ handling by overexpression of junctate in heart.n heart.

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Heterogeneity of the SR-dependent Inward $Na^+-Ca^{2+}$ Exchange Current in the Heavily $Ca^{2+}-buffered$ Rat Ventricular Myocytes

  • Yoon, Kyung-Bong;Ahn, Sung-Wan;Ko, Chang-Mann
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.2
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    • pp.101-110
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    • 2004
  • Voltage-sensitive release mechanism was pharmacologically dissected from the $Ca^{2+}-induced\;Ca^{2+}\;release$ in the SR $Ca^{2+}$ release in the rat ventricular myocytes patch-clamped in a whole-cell mode. SR $Ca^{2+}$ release process was monitored by using forward-mode $Na^+-Ca^{2+}$ exchange after restriction of the interactions between $Ca^{2+}$ from SR and $Na^+-Ca^{2+}$ exchange within micro-domains with heavy cytosolic $Ca^{2+}$ buffering with 10 mM BAPTA. During stimulation every 10 s with a pulse roughly mimicking action potential, the initial outward current gradually turned into a huge inward current of $-12.9{\pm}0.5\;pA/pF$. From the inward current, two different inward $I_{NCX}s$ were identified. One was $10\;{\mu}M$ ryanodine-sensitive, constituting $14.2{\pm}2.3%$. It was completely blocked by $CdCl_2$ (0.1 mM and 0.5 mM) and by $Na^+-depletion$. The other was identified by 5 mM $NiCl_2$ after suppression of $I_{CaL}$ and ryanodine receptor, constituting $14.8{\pm}1.6%$. This latter was blocked by either 10 mM caffeine-induced SR $Ca^{2+}-depletion$ or 1 mM tetracaine. IV-relationships illustrated that the latter was activated until the peak in $30{\sim}35\;mV$ lower voltages than the former. Overall, it was concluded that the SR $Ca^{2+}$ release process in the rat ventricular myocytes is mediated by the voltage-sensitive release mechanism in addition to the $Ca^{2+}-induced-Ca^{2+}\;release$.

Distinct Cellular Calcium Metabolism in Radiation-sensitive RKO Human Colorectal Cancer Cells

  • Kim, Yun Tai;Jo, Soo Shin;Park, Young Jun;Lee, Myung Za;Suh, Chang Kook
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.6
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    • pp.509-516
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    • 2014
  • Radiation therapy for variety of human solid tumors utilizes mechanism of cell death after DNA damage caused by radiation. In response to DNA damage, cytochrome c was released from mitochondria by activation of pro-apoptotic Bcl-2 family proteins, and then elicits massive $Ca^{2+}$ release from the ER that lead to cell death. It was also suggested that irradiation may cause the deregulation of $Ca^{2+}$ homeostasis and trigger programmed cell death and regulate death specific enzymes. Thus, in this study, we investigated how cellular $Ca^{2+}$ metabolism in RKO cells, in comparison to radiation-resistant A549 cells, was altered by gamma (${\gamma}$)-irradiation. In irradiated RKO cells, $Ca^{2+}$ influx via activation of NCX reverse mode was enhanced and a decline of $[Ca^{2+}]_i$ via forward mode was accelerated. The amount of $Ca^{2+}$ released from the ER in RKO cells by the activation of $IP_3$ receptor was also enhanced by irradiation. An increase in $[Ca^{2+}]_i$ via SOCI was enhanced in irradiated RKO cells, while that in A549 cells was depressed. These results suggest that ${\gamma}$-irradiation elicits enhancement of cellular $Ca^{2+}$ metabolism in radiation-sensitive RKO cells yielding programmed cell death.

Physiological Responses of Rice Seedlings to Butachlor (Butachlor에 대한 벼 유묘의 생리적 반응)

  • Tsai, Wen-Fu
    • Korean Journal of Weed Science
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    • v.15 no.4
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    • pp.247-253
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    • 1995
  • The herbicide butachlor [N-(butoxymethyl)-2-chloro-N-(2,6-di-methylphenyl) acetamide] is widely used by farmers as a tool for weed management of transplanted rice(Oryza sativa L.) in Taiwan. The herbicide did not stop germination of rice and weed seeds, but strongly inhibited the subsequent growth of young shoots and roots. The inhibition was also strong on established seedlings. However, they could recover to normal growth after the herbicide effect disappeared. Butachlor greatly decreased the endogenous indole-3-acetic acid (IAA) but increased the endogenous abscisic acid (ABA) contents of rice seedlings. Addition of lAA into growth medium (Hoagland's solution) partly relieved growth inhibition. Pretreatment of both gibberellic acid ($GA_3$) and IAA 24 hours before butachlor treatment almost completely alleviated the butachlor-interfere with GA and/or IAA metabolism or their action resulting in the growth inhibition of rice. Butachlor was readily absorbed by rice roots. During 24 hours of uptake experiment, 32% of the applied herbicide was absorbed. Pretreatment of the herbicide for 2 days did ncx affect the absorption. Of the absorbed herbicide, 80% remained in roots, only 20% transported into shoots, and more than 50% was metabolized to water soluble substances. Thin-layer chromatographic (TLC) analysis indicated that the Rf value of the most abundant metabolite was butachlor-glutathione conjugate. Rice, barnyardgrass (Echinochloa crus-galli (L.) Beauv.), and monochoria (Monochoria vaginalis Presl) seedlings contained relatively high level of non-protein thiols, while the glutathione S-transferase (GST) activity was found highest in rice, barnyardgrass the next, monochoria the lowest. The difference in GST activity among these species might be related to their sensitivity to butachlor.

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Intestinal segment and vitamin D3 concentration affect gene expression levels of calcium and phosphorus transporters in broiler chickens

  • Jincheng Han;Lihua Wu;Xianliang Lv;Mengyuan Liu;Yan Zhang;Lei He;Junfang Hao;Li Xi;Hongxia Qu;Chuanxin Shi;Zhiqiang Li;Zhixiang Wang;Fei Tang;Yingying Qiao
    • Journal of Animal Science and Technology
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    • v.65 no.2
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    • pp.336-350
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    • 2023
  • Two experiments were conducted in this research. Experiment 1 investigated the spatial expression characteristics of calcium (Ca) and phosphorus (P) transporters in the duodenum, jejunum, and ileum of 21-day-old broilers provided with adequate nutrient feed. Experiment 2 evaluated the effects of dietary vitamin D3 (VD3) concentration (0, 125, 250, 500, 1,000, and 2,000 IU/kg) on growth performance, bone development, and gene expression levels of intestinal Ca and P transporters in 1-21-day-old broilers provided with the negative control diet without supplemental VD3. Results in experiment 1 showed that the mRNA levels of calcium-binding protein 28-kDa (CaBP-D28k), sodium-calcium exchanger 1 (NCX1), plasma membrane calcium ATPase 1b (PMCA1b), and IIb sodium-phosphate cotransporter (NaPi-IIb) were the highest in the broiler duodenum. By contrast, the mRNA levels of inorganic phosphate transporter 1 (PiT-1) and 2 (PiT-2) were the highest in the ileum. Results in experiment 2 showed that adding 125 IU/kg VD3 increased body weight gain (BWG), feed intake (FI), bone weight, and percentage and weight of Ca and P in the tibia and femur of 1-21-day-old broilers compared with the negative control diet (p < 0.05). The rise in dietary VD3 levels from 125 to 1,000 IU/kg further increased the BWG, FI, and weights of the bone, ash, Ca, and P (p < 0.05). No difference in growth rate and leg bone quality was noted in the broilers provided with 1,000 and 2,000 IU/kg VD3 (p > 0.05). Supplementation with 125-2,000 IU/kg VD3 increased the mRNA abundances of intestinal Ca and P transporters to varying degrees. The mRNA level of CaBP-D28k increased by 536, 1,161, and 28 folds in the duodenum, jejunum, and ileum, respectively, after adding 1,000 IU/kg VD3. The mRNA levels of other Ca and P transporters (PMCA1b, NCX1, NaPi-IIb, PiT-1, and PiT-2) increased by 0.57-1.74 folds by adding 1,000-2,000 IU/kg VD3. These data suggest that intestinal Ca and P transporters are mainly expressed in the duodenum of broilers. Moreover, the addition of VD3 stimulates the two mineral transporter transcription in broiler intestines.