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Effects of 17-DMAG Administration on Autophagy Flux in Mouse Skeletal Muscle (17-DMAG이 마우스 골격근에서 autophagy flux에 미치는 영향)

  • Ju, Jeong-sun;Lee, Yoo-Hyun
    • Journal of Life Science
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    • v.26 no.4
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    • pp.387-397
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    • 2016
  • The purpose of this study was to determine if heat shock proteins are involved in autophagy in skeletal muscle. We used the autophagy flux strategy, which is an LC3 II/p62 turnover assay conducted with and without an autophagy inhibitor, to determine whether 17-DMAG (an Hsp90 inhibitor/Hsp72 activator) stimulates autophagy in skeletal muscle. We treated C2C12 cells with 17-DMAG (500 nM) for 24 hr with and without the autophagy inhibitor (Bafilomycin A1, 200 ng/ml), and we injected C57BL/6 mice i.p. with 17-DMAG (10 mg/kg) daily for 7 days with and without colchicine as an autophagy inhibitor (0.4 mg/kg/day, administered on the last 2 days). C2C12 myotubes and tibialis anterior muscles were harvested for analysis of mTOR-dependent autophagy signaling pathway proteins and autophagic marker proteins (p62 and LC3 II) by Western blot analysis. The blots showed that 17-DMAG upregulated hsp72 and decreased Akt protein levels and S6 phosphorylation in C2C12 cells. However, an in vitro autophagic flux assay demonstrated that 17-DMAG did not increase LC3 II and p62 protein concentrations to a greater extent than Bafilomycin A1 treatment alone. Similarly, 17-DMAG increased Hsp72 protein levels and decreased the expression of Akt and the phosphorylation of S6 in mouse skeletal muscle. However, unlike the response seen in C2C12 myotubes, the p62 protein levels were significantly decreased in 17-DMAG-treated mouse skeletal muscle (~50%; p<0.05). The LC3 II protein levels in 17-DMAG-treated mice were increased ~2-fold more when degradation was inhibited by colchicine (p<0.01). This suggests that 17-DMAG stimulates basal autophagy in skeletal muscle but is not found in C2C12 myotubes.

p53 Codon 72 Polymorphism Interactions with Dietary and Tobacco Related Habits and Risk of Stomach Cancer in Mizoram, India

  • Malakar, Mridul;Devi, K. Rekha;Phukan, Rup Kumar;Kaur, Tanvir;Deka, Manab;Puia, Lalhriat;Sailo, Lalrinliana;Lalhmangaihi, T.;Barua, Debajit;Rajguru, Sanjib Kumar;Mahanta, Jagadish;Narain, Kanwar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.717-723
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    • 2014
  • Background: This study was carried out to investigate the interaction of p53 codon 72 polymorphism, dietary and tobacco habits with reference to risk of stomach cancer in Mizoram, India. A total of 105 histologically confirmed stomach cancer cases and 210 age, sex and ethnicity matched healthy population controls were included in this study. Materials and Methods: The p53 codon 72 polymorphism was detected by PCR-RFLP and sequencing. H. pylori infection status was determined by ELISA. Information on various dietary and tobacco related habits was recorded with a standard questionnaire. Results: This study revealed that overall, the Pro/Pro genotype was significantly associated with a higher risk of stomach cancer (OR, 2.54; 95%CI, 1.01-6.40) as compared to the Arg/Arg genotype. In gender stratified analysis, the Pro/Pro genotype showed higher risk (OR, 7.50; 95%CI, 1.20-47.0) than the Arg/Arg genotype among females. Similarly, the Pro/Pro genotype demonstrated higher risk of stomach cancer (OR, 6.30; 95%CI, 1.41-28.2) among older people (>60 years). However, no such associations were observed in males and in individuals <60 years of age. Smoke dried fish and preserved meat (smoke dried/sun dried) consumers were at increased risk of stomach cancer (OR, 4.85; 95%CI, 1.91-12.3 and OR, 4.22; 95%CI, 1.46-12.2 respectively) as compared to non-consumers. Significant gene-environment interactions exist in terms of p53 codon 72 polymorphism and stomach cancer in Mizoram. Tobacco smokers with Pro/Pro and Arg/Pro genotypes were at higher risk of stomach cancer (OR, 16.2; 95%CI, 1.72-153.4 and OR, 9.45; 95%CI, 1.09-81.7 respectively) than the non-smokers Arg/Arg genotype carriers. The combination of tuibur user and Arg/Pro genotype also demonstrated an elevated risk association (OR, 4.76; 95%CI, 1.40-16.21). Conclusions: In conclusion, this study revealed that p53 codon 72 polymorphism and dietary and tobacco habit interactions influence stomach cancer development in Mizoram, India.

Pre-and Post-ishemic Changes of the Constituent Enzymes in Isolated Rabbit's Myocardium (허혈전후 적출 가토 심근내의 구성 효소의 변화)

  • 천수봉;전도환;이재성;김송명
    • Journal of Chest Surgery
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    • v.33 no.2
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    • pp.117-124
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    • 2000
  • Background: Nucleoside transport inhibitor(NTI) Keeps AMP, ADP, ATP levels high in myocytes by inhibiting adenosine cataboilsm so that it may preserve the myocardial contractability during ischemia In this study we investigated the effects of cyclic AMP phosphodiesterase inhibor(C-AMP PDSI) and S-P-nitrobenzyl-6 -thioniosine(NBT; a sort of NIT) on myocadial preservation and changes of constituent enzyme. Material and method: Twenty-six isolated rabbit hearts were perfused with Krebs-Henseleit buffer solution for 20 minutes arrested for 20 minutes and ten reperfused for 30 minutes. The following four groups were prepared and hemodynamic changes coronary effluent lactate dehydrogenase (LDH) a-hydroxybutylic accid(a-HBD) levels and myocardial LDH creatine kinase-MB (CK-MB) adenosine deaminase(ADA) a-HBD levels and myocardial LDH creatine kinase-MB (CK-MB) adenosine deaminase(ADA) a-HBD levels were analysed before and after cardiac arest ; Group I(control) ; the heart was only perfused with K-H ; Group II ; the heart was perfused with K-H including C-AMP PDSI(Amrinone 25mg/L); Group III ; the heart was perfused with K-H including NBT(4.19mg/L) ; Group IV ; the heart was perfused with K-H including C-AMP PDSI + NBT. Result : Left venticular developed pressure(LVDP) at 10 minutes of the equilibrium was significantly higher in group III(72.1$\pm$5.3 mmHg p<0.01) and group III(72$\pm$5.6 mmHg P<0.025) as compared with group I (40.8$\pm$4.7mmHg) and LVDP at 20 minutes of the reperfusion was significantly higher in group II(74$\pm$5.3mmHg p<0.01) and group III(72$\pm$5.6mmHg p<0.025) as compared with group I (44.2$\pm$4.6mmHg). Percentage recovery of LVDP at the reperfusion was the highest in group II(123.3%) Percentage recovery of coronary flow at the equilibrium reperfusion were higher in group II(310%, 270%) group III(230%, 290%) group IV(310%, 280%) as compared with group I (100%) respectively. Myocadial LDH level was significant lower in group IV(33495$\pm$1802 IU/gm p<0.04) as compared with group I(48767$\pm$1421 IU/gm) Myocadial CK-MB level was significant higher in group II(74820$\pm$1421 IU/gm) compared with group I (45450$\pm$1737 IU/gm) Myocadial ADA level was significant higher group IV(1215$\pm$8 IU/gm p<0.05) compared with group I(125$\pm$15 IU/gm) but there was no significant difference between group I and group II ,III, IV in changes of coronary effluent LDH, a-HBD levels. Conclusion: C-AMP PDSI solely appears to have a better effect on myocardial preservation after ischemia than NBT but with no synergistic effect and it could keep CK-MB leve high in myocardial tissues.

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Effects of the Duration of Liquid Feeding on Performance and Nutrient Digestibility in Weaned Pigs

  • Han, Yung-Keun;Thacker, P.A.;Yang, Joo-Sung
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.396-401
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    • 2006
  • A total of 72 castrated, crossbred ($Landrace{\times}Yorkshire$) piglets ($5.7{\pm}0.7kg$ BW) were allotted to one of three treatments including: a dry crumbled feed fed for 40 days; liquid feed provided for 10 days followed by dry crumbled feed for 30 days; and liquid feed provided for 20 days followed by dry crumbled feed for 20 days. Liquid feed was produced fresh each day by mixing water with dry crumbled feed at a ratio of 3:1. Pigs fed liquid feed for 10 days had a higher weight gain during the first 10 days (+16.8%; p = 0.01) and over the entire experimental period (+4.9%; p = 0.07) than pigs offered dry feed. Pigs fed liquid feed for 20 days also had a higher weight gain during the period from d 0 to 10 (+12.8%; p = 0.01), from d 10 to 20 (+8.5%; p = 0.06) and from d 0 to 20 (+9.7%; p = 0.01) than pigs offered dry feed. Pigs fed liquid feed for the first 10 days had higher feed intakes from d 0 to 10 (+22.6%; p = 0.01) and from d 0 to 40 (+5.3%; p = 0.02) than pigs offered dry feed. Pigs fed liquid feed for the first 20 days had a higher feed intake from d 0 to 10 (+21.8%; p = 0.01), from d 10 to 20 (+10.6%; p = 0.06), from d 0 to 20 (+14.6%; p = 0.01) and from d 0 to 40 (+6.6%; p = 0.02) than pigs offered dry feed. Feed conversion from d 0 to 40 tended to be poorer for pigs fed liquid feed during the first 20 days (p = 0.08) indicating an increase in feed wastage with liquid feeding. Nutrient digestibility at day 10 was unaffected by dietary treatment. However, at day 30, pigs fed liquid feed for 10 days had higher digestibility of dry matter (p = 0.08), energy (p = 0.10), crude protein (p = 0.03) and neutral detergent fibre (p = 0.05) than pigs fed liquid feed for 20 days. In conclusion, liquid feeding for 10 or 20 days increased the performance of weaned pigs during the specific time period that liquid feeding occurred but there were no carry over effects into subsequent production periods.

Analisis of chief influencing factor on aged man's cerebrovascular accidents - case control study - (노인 중풍 발생에 미치는 요인 분석)

  • Choi, Young-Mi;Ryu, Tae-Sik;Lee, Jong-Sub
    • Journal of agricultural medicine and community health
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    • v.23 no.1
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    • pp.51-64
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    • 1998
  • This study was carried out during 3 years from March 1992 to March 1995, 140 healthy people and 140 C.V.A patients were investigated, at an oriental clinic in Taejon. The data collected was analysed using a SAS. The results are as follows. 1. The Relationships between patients who have undergone one or more abortion procedures and C.V.A occurrence. A statistical significant difference was recorded between the control group and the outpatients group of P<0.05 2. Number of years after initial C.V.A diagnosis before C.V.A symptom onset. within 1 years 52% (2-3 years, 3-4 years, 4-5 years, 5 years and over) 3. Additional diseases occurring after initial stroke 1 disease 50.7% (2, 3, 4, 5 or more) 4. Alcohol Consumption "A statistical significant difference was recorded between the control group and the outpatients group of" P<0.01 5. Previous Treatment Experience for stoke outpatients Oriental Hospital, clinic 64.3% (General Hospital, clinic, a public health center drug store, home remedies, etc) 6. Exercise Frequency Outpatients Regular 28.6%, Irregular 71.4% Control sample Regular 27%, Irregular 72.9% 7. Degree of Appetite "A statistical significant difference was recorded between the control group and the outpatients group of" P<0.01 8. People who include fish in their diet "A statistical difference was recorded between the control group and the outpatients group of" P<0.01.

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METABOLIZABLE ENERGY REQUIREMENTS FOR MAINTENANCE AND GROWTH OF SUCKLING CALVES GIVEN MILK REPLACER

  • Sekine, J.;Oura, R.;Morita, Z.;Morooka, T.;Asahida, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.1 no.2
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    • pp.61-63
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    • 1988
  • A total of 38 energy balance trials were done for calves fed a liquid milk replacer, calf starter and second cut mixed hay. Milk replacer supplied 81% of the total dietary energy. Live weight of calves averaged 54.1 (S.D 6.2) kg and daily gain 0.37 (${\pm}0.23$)kg. The metabolizability of gross energy averaged 0.822. A regression was calculated relating energy retention (ER, $kJ/kg^{0.75}$) to the intake of metabolizable energy (IME, $kJ/kg^{0.75}$): ER = 0.72 (${\pm}0.12$) IME - 330, r = 0.702, P < 0.01, $S.E.{\pm}18.0$. Metabolizable energy for maintenance (MEm) was calculated to be $458kJ/kg^{0.75}$ when ER = 0. The amount of IME over MEm for an individual animal (Meg, $kJ/kg^{0.75}$) was regressed on averaged daily gain (ADG, kg): Meg = 413 (${\pm}91$) ADG + 0.2, r = 0.650, P < 0.01, $S.E.{\pm}21$. The amount of ME requirement for suckling calves was estimated using values obtained above.

A Comparision of the Limits of Stability at Different Sensory Conditions in 20 Years of Age (20대 연령에서 다양한 감각 조건에 따른 안정성 한계의 비교)

  • Kwon, Oh-Yun;Choi, Houng-Sik
    • Journal of Korean Physical Therapy Science
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    • v.3 no.2
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    • pp.963-973
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    • 1996
  • The purpose of this study was to evaluate and compare the limits of stability(LOS) at different sensory conditions in normal 20 years of age. The LOS was measured at stable surface, and unstable surface and the subjects stood with the feet contacted and 4 inches between the feet with the eyes open and the eyes closed. In this study, 20 physical therapy major subjects were evaluated at Wonkwang Public Health Junior College. In this study applied the paired t-test, and Kruskal-Wallis 1-way ANOVA to determine the statistical significance of results at 0.01 level of significance. The results were as follows: 1. The mean of lateral limits of stability was b.67 degree at stable surface with the eyes open and standing with the feet contacted. 2. The mean of anteroposterior limits of stability was 9.78 degree at stable surface with the eyes open and standing with the feet contacted. 3. The mean of lateral limits of stability was 15.10 degree at stable surface with the eyes open and standing with 4 inches between the feet. 4. The mean of anteroposterior limits of stability was 11.72 degree at stable surface with the eyes open and standing with 4 inches between the feet. 5. The anterior-posterior and lateral limits of stability significantly decreased with the eyes closed(p<0.01). 6. The anterior-posterior and lateral limits of stability significantly decreased at unstable surface(p<0.01). 7. There was no significant difference of limits of stability as the height and foot length(p>0.01).

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Effect of Benzalkonium Chloride on Biogas Potential of Pig Slurry (Benzalkonium Chloride가 돈슬러리의 바이오가스 생성에 미치는 영향)

  • Park, H. R.;Choi, H. L.;Suresh, Arumuganainar
    • Journal of the Korea Organic Resources Recycling Association
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    • v.19 no.3
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    • pp.63-72
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    • 2011
  • Benzalkonium chloride is most widely used in S. Korea as a disinfectant in livestock husbandry. Inhibition of biogas potentials were investigated with three different doses of benzalkonium chloride in swine slurry. The system was operated at batch mode. The inhibition rates were 10%, 30~40% and >70% at the dose of 10ppm, 40ppm and 80ppm, respectively assuming it was zero percent in case of no dose. Enzymatic activities were analyzed to determine the enzymatic type which was inhibited by benzalkonium chloride. The acid phosphatase, alkaline phosphatase and protease were shown negatively correlated with biogas potential. Correlation of ${\alpha}$-glucosidase and biogas potentials was observed not high (p<0.01, r=-0.426) while benzalkonium chloride (r=-0.853, p<0.01) and acid phosphatase (p<0.01) with biogas potentials were significantly and negatively correlated. The effect of benzalkonium chloride on Escherichia coli were also evaluated by disc diffusion method. As increase of benzalkonium concentration, inhibition zone of anaerobic bacteria was extended. It revealed that benzalkonium chloride significantly deteriorated biogas potential through inhibition of acetogenic bacteria.

The effect of eleutherococcus senticosus on metabolism-associated protein expression in 3T3-L1 and C2C12 cells

  • Hashimoto, Takeshi;Okada, Yoko;Yamanaka, Atsushi;Ono, Natsuhiko;Uryu, Keisuke;Maru, Isafumi
    • Korean Journal of Exercise Nutrition
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    • v.24 no.3
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    • pp.13-18
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    • 2020
  • [Purpose] In vivo studies have demonstrated the ergogenic benefits of eleutherococcus senticosus (ES) supplementation. ES has been observed to enhance endurance capacity, improve cardiovascular function, and alter metabolic functions (e.g., increased fat utilization); however, the exact mechanisms involved remain unknown. We aimed to determine whether ES could effectively induce fat loss and improve muscle metabolic profiles through increases in lipolysis- and lipid metabolism-associated protein expression in 3T3-L1 adipocytes and C2C12 skeletal muscle cells, respectively, to uncover the direct effects of ES on adipocytes and skeletal muscle cells. [Methods] Different doses of ES extracts (0.2, 0.5, and 1.0 mg/mL) were added to cells (0.2 ES, 0.5 ES, and 1.0 ES, respectively) for 72 h and compared to the vehicle control (control). [Results] The intracellular triacylglycerol (TG) content significantly decreased (p < 0.05 for 0.2 ES, p < 0.01 for 0.5 ES and 1.0 ES) in 3T3-L1 cells. Adipose triglyceride lipase, which is involved in active lipolysis, was significantly higher in the 1.0 ES group than in the control group (p < 0.01) of 3T3-L1 adipocytes. In C2C12 cells, the mitochondrial protein voltage-dependent anion channel (VDAC) was significantly increased in the 1.0 ES group (p < 0.01). Furthermore, we found that 1.0 ES activated both 5' AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) in skeletal muscle cells (p < 0.01). [Conclusion] These findings suggest that ES extracts decreased TG content, presumably by increasing lipase in adipocytes and metabolism-associated protein expression as well as mitochondrial biogenesis in muscle cells. These effects may corroborate previous in vivo findings regarding the ergogenic effects of ES supplementation.