• Title/Summary/Keyword: GTP

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Inhibitory Effect of Caffeine on Carbachol-Induced Nonselective Cationic Current in Guinea-Pig Gastric Myocytes

  • Kim, Sung-Joon;Min, Kyung-Wan;Kim, Young-Chul;Lee, Sang-Jin;So, In-Suk;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.3
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    • pp.353-359
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    • 1998
  • In gastrointestinal smooth muscle, muscarinic stimulation by carbachol (CCh) activates nonselective cation channel current ($I_{CCh}$) which is facilitated by intracellular [$Ca^{2+}$] increase. Caffeine is widely used in experiments to mobilize $Ca^{2+}$ from intracellular stores. This study shows a strong inhibitory effect of caffeine on $I_{CCh}$ in guinea-pig gastric myocyte. In this study, the underlying mechanism of the inhibitory effect of caffeine was investigated. $I_{CCh}$ was completely suppressed by the addition of caffeine (10 mM) to the superfusing solution. Inhibition of $I_{CCh}$ by caffeine was not related to the intracellular cAMP accumulation which was expected from the phosphodiesterase-inhibiting effect of caffeine. The blockade of $InsP_3-induced$ $Ca^{2+}$ release by heparin had no significant effects on the activation of $I_{CCh}$. When the same cationic current had been induced by intracellular dialysis of $GTP[{\gamma}S]$ in order to bypass the muscarinic receptor, the inhibitory effect of caffeine was significantly attenuated. The results of this study indicate that both intracellular signalling pathways for $I_{CCh}$, proximal and distal to G-protein activation, are suppressed by caffeine. A major inhibition was observed at the proximal level.

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Relationship of the Signal Transduction-mediated Proteins and Enzymes to Contractility and Plasticity in Skeletal Muscles (골격근의 수축과 가소성에 대한 신호전달-매개 단백질 및 관련 효소의 상관성)

  • Kim, Jung-Hwan
    • The Journal of Korean Physical Therapy
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    • v.19 no.4
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    • pp.1-14
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    • 2007
  • Background: It is generally accepted that skeletal muscle contraction is triggered by nerve impulse and intracellular $Ca^{2+}\;([Ca^{2+}]_i)$ released from intracellular $Ca^{2+}$ stores such as sarcoplasmic reticulum (SR). Specifically, this process, called excitation-contraction (E-C) coupling, takes place at intracellular junctions between the plasma membrane, the transverse (T) tubule L-type $Ca^{2+}$ channel (dihydropyridine-sensitive L-rype $Ca^{2+}$ channel, DHPR, also called tetrads), and the SR $Ca^{2+}$ release channel (ryanodine-sensitive $Ca^{2+}$ release channel, RyR, also called feet) of internal $Ca^{2+}$ stores in skeletal muscle cells. Furthermore, it has been reported that the $Ca^{2+-}$ dependent and -independent contraction determine the expression of skeletal muscle genes, thus providing a mechanism for tightly coupling the extent of muscle contraction to regulation of muscle plasticity-related excitation-transcription (E-T) coupling. Purpose: Expression and activity of plasticity-associated enzymes in gastrocnemius muscle strips have not been well studied, however. Methods: Therefore, in this study the expression and phosphorylation of E-C and E-T coupling-related mediators such as protein kinases, ROS(reactive oxygen species)- and apoptosis-related substances, and others in gastrocnemius muscles from rats was examined. Results: I found that expression and activity of MAPKs (mitogen-activated protein kinases, ERK1/2, p38MAPK, and SAPK/JNK), apoptotic proteins (cleaved caspase-3, cytochrome c, Ref-1, Bad), small GTP-binding proteins (RhoA and Cdc42), actin-binding protein (cofilin), PKC (protein kinase C) and $Ca^{2+}$ channel (transient receptor potential channel 6, TRPC6) was observed in rat gastrocnemius muscle strips. Conclusion: These results suggest that MAPKs, ROS- and apoptosis-related enzymes, cytoskeleton-regulated proteins, and $Ca^{2+}$ channel may in part functionally import in E-C and E-T coupling from rat skeletal muscles.

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Effects of scalp treatment using combinational massage technique on human physiology

  • Oh, Gang-Su;Kim, Sung-Nam
    • Journal of Fashion Business
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    • v.12 no.3
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    • pp.87-98
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    • 2008
  • The purposes of this study are to measure physiological reactions of human body according to the scalp treatment, a popularized service in beauty care industry and propose efficient ways of scalp treatment. To meet the goals, total 30 applicants without any medical history(5 males and 5 females in 20's, 30's and 40's respectively) were informed on the purpose of experiment hereof and were investigated and received a 30-minute scalp treatment, which combines standardized scalp treatment massage technique proposed by KAT and ITF with another massage technique operated in the beauty salon run by the author of this paper. 5ml of blood samples were taken from each subject before and after the scalp treatment respectively and the blood sample was divided into 3 different tubes for analysis: 1) 2 ml for blood cell analysis, 2) 2ml for enzyme activity measurement, 3) 1ml for hormone level reading. In order to determine effects of scalp treatment on ALP, GOT, GPT, ${\gamma}-GTP$, WBC, RBC, Hb, Hct, Platelet, MCV, MCH and MCHC, all collected data were used for measuring respective levels of these blood substances by means of enzyme reaction measurement, enzyme activity measurement and automated hematology analyzer. Then, all measured data were analyzed through paired t-test using SPSS WIN 11.5. As a result, the scalp treatment is associated with improving hepatic function, facilitating blood circulation and helping blood coagulation and hemostasis in a effective way. Therefore, it would be necessary to conduct further studies on this subject related to anemia in the future.

The Dietary Intake and Nutritional Status of Fatty Liver Patients (지방간 환자의 식품섭취 및 영양상태에 관한 조사-남성을 중심으로-)

  • 이석화;기춘석;장유경
    • Korean Journal of Community Nutrition
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    • v.3 no.4
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    • pp.622-629
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    • 1998
  • This study was performed to investigate the effect of dietary intakes and nutritional status for fatty liver male patients with fatty livers. General characteristics, nutritional knowledge, and dietary intakes were obtained from fatty liver male patients aged 20-60yr living in Seoul and Kyunggi province by individual interview and blood samples. The Results were as follows : the mean age of the subjects was 39yrs. Old and over half the subjects were overweight. Although nutritional knowledge, recognition, and accuracy scores were good, these did not affect the subject's health. The average alcohol intake of subjects was 79.8g/day and the drinking duration was 5-40yrs(average : 17yrs). The subjects drank alcohol over 4tiles/week and preferred Soju. Most side dishes taken with alcohol drinking by the subjects were foods from animal sources. Half of the subjects also drank alcohol the next day for breakfast as a hangover-chaser. The reasons for drinking were due to social relationships and by habit. Vitamin D and folate were lower than the RDA for Koreans, and other nutrient amounts were higher than the RDA. The amounts of aspartate amino-transferase(AST), Alanine amino-transferase(ALT), ${\gamma}$-glutamyl transpeptidase(${\gamma}$-GTP), Triglyceride(TG) for the subjects were higher. In conclusion, fatty liver patients drank excessively and frequently. It seems that heavy alcohol drinking is more prone to overweight or obese people and contributes to fatty livers. Therefore, the subjects need to be taken effectively educated about alcohol-induced liver damage and the importance of a balanced diet for on healthy liver.

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Effects of Fly Maggot Extracts on the Liver and Plasma Lipid in Rat Fed High-Fat Diets (고지방식이 유도된 흰쥐의 혈액지질 및 간에 관한 파리유충 추출물의 효과)

  • Park, Byung-Sung
    • Journal of the Korean Applied Science and Technology
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    • v.27 no.3
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    • pp.290-299
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    • 2010
  • The bioactive effects of ethanol extracts from fly maggot (ME) on reduction of plasma lipids levels in rats fed high-fat diets (Expt. Ⅰ), and on liver function recovery of hepatotoxicity rats by intraperitoneal injection of carbon tetrachloride ($CCl_4$) or by orally administration of alcohol (Expt. II) were investigated. In expt. I, twenty seven, male rat SDS(sprague dawley strain) were randomly assigned to three treated groups, including normal control group, HF (group with high fat diets which have no extracts) and HFE (HF plus orally administered doses of ME extract at 5.0 mg/100g of body weight). In expt. II, forty five, male rats (SDS) were randomly assigned to each of the five groups: T1 (control), T2 (intraperitoneal injection of $CCl_4$), T3 (intraperitoneal injection of $CCl_4$ after orally administered with ME), T4 (orally administered with combination of ME and alcohol), T5 (orally administration of ME after orally administered with alcohol). There were significant decreases in plasma (TAG), (TC), (LDL-C) in the HFE group with orally administered doses of ME at 5.0 mg/100g of body weight, respectively, however, the (HDL-C) were significantly increased in HFE group as compared to HF group with high fat diets which have no extracts (p<0.05). The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), ${\gamma}$-glutamyl transferse(${\gamma}$-GTP) and bilirubin were highest in T2 or T3, and high in order T4 or T5, and lowest in T1 except for bilirubin which has same with T4, T5 (p<0.05). The high recovery of liver damage by $CCl_4$ from the light microscopic appearance was observed in rats (T3) with extracts, and also high in T4 than T5 by orally administrated with alcohol. In conclusion, the ethanol extracts from fly maggot may have a bioactive effects to prevent for human lipids disorder and alcoholic disease.

Identification of phospholipase Cβ downstream effect on transient receptor potential canonical 1/4, transient receptor potential canonical 1/5 channels

  • Ko, Juyeon;Myeong, Jongyun;Kwak, Misun;Jeon, Ju-Hong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.5
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    • pp.357-366
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    • 2019
  • $G{\alpha}_q$-coupled receptor stimulation was implied in the activation process of transient receptor potential canonical (TRPC)1/4 and TRPC1/5 heterotetrameric channels. The inactivation occurs due to phosphatidylinositol 4,5-biphosphate ($PI(4,5)P_2$) depletion. When $PI(4,5)P_2$ depletion was induced by muscarinic stimulation or inositol polyphosphate 5-phosphatase (Inp54p), however, the inactivation by muscarinic stimulation was greater compared to that by Inp54p. The aim of this study was to investigate the complete inactivation mechanism of the heteromeric channels upon $G{\alpha}_q$-phospholipase $C{\beta}$ ($G{\alpha}_q-PLC{\beta}$) activation. We evaluated the activity of heteromeric channels with electrophysiological recording in HEK293 cells expressing TRPC channels. TRPC1/4 and TRPC1/5 heteromers undergo further inhibition in $PLC{\beta}$ activation and calcium/protein kinase C (PKC) signaling. Nevertheless, the key factors differ. For TRPC1/4, the inactivation process was facilitated by $Ca^{2+}$ release from the endoplasmic reticulum, and for TRPC1/5, activation of PKC was concerned mostly. We conclude that the subsequent increase in cytoplasmic $Ca^{2+}$ due to $Ca^{2+}$ release from the endoplasmic reticulum and activation of PKC resulted in a second phase of channel inhibition following $PI(4,5)P_2$ depletion.

Englerin A-sensing charged residues for transient receptor potential canonical 5 channel activation

  • Jeong, SeungJoo;Ko, Juyeon;Kim, Minji;Park, Ki Chul;Park, Eunice Yon June;Kim, Jinsung;Baik, Youngjoo;Wie, Jinhong;Cho, Art E.;Jeon, Ju-hong;So, Insuk
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.3
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    • pp.191-201
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    • 2019
  • The transient receptor potential canonical (TRPC) 5 channel, known as a nonselective cation channel, has a crucial role in calcium influx. TRPC5 has been reported to be activated by muscarinic receptor activation and extracellular pH change and inhibited by the protein kinase C pathway. Recent studies have also suggested that TRPC5 is extracellularly activated by englerin A (EA), but the mechanism remains unclear. The purpose of this study is to identify the EA-interaction sites in TRPC5 and thereby clarify the mechanism of TRPC5 activation. TRPC5 channels are over-expressed in human embryonic kidney (HEK293) cells. TRPC5 mutants were generated by site-directed mutagenesis. The whole-cell patch-clamp configuration was used to record TRPC5 currents. Western analysis was also performed to observe the expression of TRPC5 mutants. To identify the EA-interaction site in TRPC5, we first generated pore mutants. When screening the mutants with EA, we observed the EA-induced current increases of TRPC5 abolished in K554N, H594N, and E598Q mutants. The current increases of other mutants were reduced in different levels. We also examined the functional intactness of the mutants that had no effect by EA with TRPC5 agonists, such as carbachol or $GTP{\gamma}S$. Our results suggest that the three residues, Lys-554, His-594, and Glu-598, in TRPC5 might be responsible for direct interaction with EA, inducing the channel activation. We also suggest that although other pore residues are not critical, they could partly contribute to the EA-induced channel activation.

Engineering of Biosynthesis Pathway and NADPH Supply for Improved L-5-Methyltetrahydrofolate Production by Lactococcus lactis

  • Lu, Chuanchuan;Liu, Yanfeng;Li, Jianghua;Liu, Long;Du, Guocheng
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.154-162
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    • 2021
  • L-5-methyltetrahydrofolate (5-MTHF) is one of the biological active forms of folate, which is widely used as a nutraceutical. However, low yield and serious pollution associated with the chemical synthesis of 5-MTHF hampers its sustainable supply. In this study, 5-MTHF production was improved by engineering the 5-MTHF biosynthesis pathway and NADPH supply in Lactococcus lactis for developing a green and sustainable biosynthesis approach. Specifically, overexpressing the key rate-limiting enzyme methylenetetrahydrofolate reductase led to intracellular 5-MTHF accumulation, reaching 18 ㎍/l. Next, 5-MTHF synthesis was further enhanced by combinatorial overexpression of 5-MTHF synthesis pathway enzymes with methylenetetrahydrofolate reductase, resulting in 1.7-fold enhancement. The folate supply pathway was strengthened by expressing folE encoding GTP cyclohydrolase I, which increased 5-MTHF production 2.4-fold to 72 ㎍/l. Furthermore, glucose-6-phosphate dehydrogenase was overexpressed to improve the redox cofactor NADPH supply for 5-MTHF biosynthesis, which led to a 60% increase in intracellular NADPH and a 35% increase in 5-MTHF production (97 ㎍/l). To reduce formation of the by-product 5-formyltetrahydrofolate, overexpression of 5-formyltetrahydrofolate cyclo-ligase converted 5-formyltetrahydrofolate to 5,10-methyltetrahydrofolate, which enhanced the 5-MTHF titer to 132 ㎍/l. Finally, combinatorial addition of folate precursors to the fermentation medium boosted 5-MTHF production, reaching 300 ㎍/l. To the best of our knowledge, this titer is the highest achieved by L. lactis. This study lays the foundation for further engineering of L. lactis for efficient 5-MTHF biosynthesis.

The Small GTPase CsRAC1 Is Important for Fungal Development and Pepper Anthracnose in Colletotrichum scovillei

  • Lee, Noh-Hyun;Fu, Teng;Shin, Jong-Hwan;Song, Yong-Won;Jang, Dong-Cheol;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.607-618
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    • 2021
  • The pepper anthracnose fungus, Colletotrichum scovillei, causes severe losses of pepper fruit production in the tropical and temperate zones. RAC1 is a highly conserved small GTP-binding protein in the Rho GT-Pase family. This protein has been demonstrated to play a role in fungal development, and pathogenicity in several plant pathogenic fungi. However, the functional roles of RAC1 are not characterized in C. scovillei causing anthracnose on pepper fruits. Here, we generated a deletion mutant (𝜟Csrac1) via homologous recombination to investigate the functional roles of CsRAC1. The 𝜟Csrac1 showed pleiotropic defects in fungal growth and developments, including vegetative growth, conidiogenesis, conidial germination and appressorium formation, compared to wild-type. Although 𝜟Csrac1 was able to develop appressoria, it failed to differentiate appressorium pegs. However, 𝜟Csrac1 still caused anthracnose disease with significantly reduced rate on wounded pepper fruits. Further analyses revealed that 𝜟Csrac1 was defective in tolerance to oxidative stress and suppression of host-defense genes. Taken together, our results suggest that CsRAC1 plays essential roles in fungal development and pathogenicity in C. scovilleipepper fruit pathosystem.

Physicochemical and Antioxidant Properties of Sponge Cake made using Barley Sprout and Green Tea (새싹보리와 녹차 스펀지케이크의 이화학적 및 항산화적 품질 특성)

  • Kim, Eunkyung
    • Journal of the Korean Society of Food Culture
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    • v.37 no.1
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    • pp.90-98
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    • 2022
  • This study investigated the physicochemical and antioxidant properties of sponge cakes made using different amounts (2 and 4%) of barley sprout powder (BSP) and green tea powder (GTP), respectively. The results showed that the baking loss rate of GS2 (2% green tea) and GS4 (4% green tea) was 12.39% and 11.96%, respectively in the green tea addition group, which was higher than that of the barley sprout group, but significantly lower than that of the control group at 13.34% (p<0.05). The specific volumes of the sponge cake containing barley sprout and green tea 2% and 4% were between 2.84-2.95 mL/g, which was significantly higher compared to the control group at 2.69 mL/g (p<0.05). The sugar content was significantly higher in the control group and the barley sprout addition group at 2.30°Brix (p<0.05). As for the volume index, the control group with the lowest value at 13.43 showed a significant difference compared to the addition groups. The volume index significantly decreased as the addition amount increased, measuring 14.07 in BS4 (4% BSP) compared to 14.87 of BS2 (2% BSP) in the barley sprout group (p<0.05). In cross-sectional photography, the color became darker than that of the control group as the quantum of additives increased. In terms of the DPPH and ABTS radical scavenging activity, the total phenol content, and total flavonoid content, the groups with the addition of barley sprout and green tea showed higher antioxidant activity than the control group (p<0.05).