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Identification of Higenamine nad its Metabolites in Rat by Gas Chromatography/Mass Spectrometry

  • Ryu, Jae-chun;Song, Yun-Seon;Kim, Myung-Soo;Cho, Jung-Hyuck;Yunchoi, Hye-Sook
    • Archives of Pharmacal Research
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    • v.16 no.3
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    • pp.213-218
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    • 1993
  • ($\pm$)-Higenamine is known as a cardiotonic principle of aconite root (root of Aconitum spp., Ranunculaceae). A simple and sensitive detection method for higenamine was developed by using gas chromatography-mass spectrometry (GC/MS). The recovery of higenamine after extraction and concentration with XAD-2 resin column was around 95% from rat biological fluids such as bile, plasma and urine. The limits of detection of higenamine in these biological fluids were approximately 0.1 ng/ml each. It has well been suggested that tetrahydroisoquinolines possessing catechol moiety such as higenamine should be subjected to the catechol-O-methyl transferase (COMT) activity in vivo. We detected two major peaks of presumed metabolites of higenamine in the total ion chromatogram obtained from the rat urine sample after the oral adminstration of ($\pm$)-higenamine. The scan mass spectrum of one of the metabolties coincided with those obtained from coclaurine $(C_6$-O-methyl higenamine) and those of the other metabolite are suggestive of isococlaurine $(C_7$-O-methyl higenamine).

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Microbial Control of Forest Insect Pests (II) (산림해충의 미생물적 방제 2)

  • 이응래;황계성
    • Korean Journal of Microbiology
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    • v.9 no.2
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    • pp.69-73
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    • 1971
  • On June in 1970 the authors discovered a pathogenecity, cytoplasmic polyhedrosis virus, of the Smithia virus in the larvae of Liparis dispay L. appeared on quercus forest in Chung-Neung district and had carried out a experiment to detect the pathogenecity of Smithia virus through the inoculation of it into the larvase, such as Liparis dispay L. Hyphantrea cunea DRURY, and Dendrolinus spectabilis BUTLER. The results obtained were as follows ; 1) Death rate of L.dispay and D.spectabilis treated by 10$^{6}$ /ml cytoplasmic polyhedrosis virus of Smithis virus were 88.0% and 85.5% respectively, when the larvaes of these insects are big enough. But there were none of pathogenecity in case of Hyphantrea cunea DRURY. 20 Dead larvae caused by the injection of Smithia virus had begum to find out about on 10 days after inoculation. Miximum death rate of L. didpay and D. spectabilis appeared on 20-25days nad on 25-30days, respectively, after the incoulation. 3) In the cytoplasm of Mid-gut cylindrical cells of both of these insects, polyhedrosis, such s hexagonal (0.5-2.0-6.0 micron) were found out and in these insects, polyhedrosis, such as hexaginal (0.5-2.0-6.0 mivton) were found out and in case of D.spectabilis were a few polyhedrosis, such as tetragonal, trianglar polyhedrosis. 4) Diluted concentration of `0$^{6}$ /ml cytoplasmic polyhedrosis virus of Smithia virus were spread out in the field conditions. The corrected mortality was confirmed as about 87.8%.

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Cellular and Biochemical Alterations in L6 Myoblast Cells Induced by 6-Aminonicotinamide

  • Jang, Min-Young;Kim, Sun-Jung;Shin, Sook;Park, In-Kook
    • Animal cells and systems
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    • v.11 no.1
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    • pp.17-22
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    • 2007
  • The effects of antimetabolite 6-AN (6-amino-nicotinamide) on viability and morphology of L6 myoblast cells have been investigated. 6-AN ($100{\mu}M$) induced a time-dependent decrease in cell viability with respect to the untreated control cells. Following 6-AN administration the viability rate started to decline sharply, reaching about 23% of the untreated control cells at 48 h. Inverted phase-contrast microscopy revealed that 6-AN caused characteristic morphological changes such as irregularly elongated and stellate shape of cells, round-shaped nucleus, cytoplasmic vacuolization, irregular cell arrangements and formation of large spaces among cell clusters. The concentrations of ATP and $NAD^{+}$ in the 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. In contrast, the concentration of AMP was significantly increased by the 6-AN treatment. Activities of catalase, superoxide dismutase and glutathione peroxidase in 6-AN treated cells were significantly higher (p < 0.01) than those of the untreated control cells. The activities of glyceraldehyde-3-phosphate dehydrogenase in 6-AN treated cells were significantly lower (p < 0.01) than those of the untreated control cells. The results suggest that 6-AN caused marked reduction of cell viability and alterations of some important metabolites and enzymes.

Protective effects of 5-aminolevulinic acid on heat stress in bovine mammary epithelial cells

  • Islam, Md Aminul;Noguchi, Yoko;Taniguchi, Shin;Yonekura, Shinichi
    • Animal Bioscience
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    • v.34 no.6
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    • pp.1006-1013
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    • 2021
  • Objective: Cells have increased susceptibility to activation of apoptosis when suffering heat stress (HS). An effective supplementation strategy to mimic heat-induced apoptosis of bovine mammary epithelial cells (MECs) is necessary to maintain optimal milk production. This study aimed to investigate possible protective effects of the anti-apoptotic activity of 5-aminolevulinic acid (5-ALA) against HS-induced damage of bovine MECs. Methods: Bovine MECs were pretreated with or without 5-ALA at concentrations of 10, 100, and 500 µM for 24 h followed by HS (42.5℃ for 24 h and 48 h). Cell viability was measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Real-time quantitative polymerase chain reaction and Western blotting were used to explore the regulation of genes associated with apoptosis, oxidative stress, and endoplasmic reticulum (ER) stress genes. Results: We found that 5-ALA induces cytoprotection via inhibition of apoptosis markers after HS-induced damage. Pretreatment of bovine MECs with 5-ALA resulted in dramatic upregulation of mRNA for nuclear factor erythroid-derived 2-like factor 2, heme oxygenase-1, and NAD(P)H quinone oxidoreductase 1, all of which are antioxidant stress genes. Moreover, 5-ALA pretreatment significantly suppressed HS-induced ER stress-associated markers, glucose-regulated protein 78, and C/EBP homologous protein expression levels. Conclusion: 5-ALA can ameliorate the ER stress in heat stressed bovine MEC via enhancing the expression of antioxidant gene.

Functionalized Poplar Powder as a Support Material for Immobilization of Enoate Reductase and a Cofactor Regeneration System

  • Li, Han;Cui, Xiumei;Zheng, Liangyu
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.607-616
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    • 2019
  • In this study, functionalized poplar powder (FPP) was used as a support material for the immobilization of enoate reductase (ER) and glucose-6-phosphate dehydrogenase (GDH) by covalent binding. Under optimal conditions, the immobilization efficiency of ER-FPP and GDH-FPP was 95.1% and 84.7%, and the activity recovery of ER and GDH was 47.5% and 37.8%, respectively. Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) analysis indicated that FPP was a suitable carrier for enzyme immobilization. ER-FPP and GDH-FPP exhibit excellent thermal stabilities and superior reusability. Especially, ER-FPP and GDH-FPP enable the continuous conversion of 4-(4-Methoxyphenyl)-3-buten-2-one with $NAD^+$ recycling. While the immobilization strategies established here were simple and inexpensive, they exploited a new method for the immobilization and application of ER and its cofactor recycling system.

Oral administration of ginseng berry concentrate improves lactate metabolism and increases endurance performance in mice

  • Eun-Ju Jin;Shibo Wei;Yunju Jo;Thanh T. Nguyen;Moongi Ji;Man-Jeong Paik;Jee-Heon Jeong;Se Jin Im;Dongryeol Ryu
    • BMB Reports
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    • v.56 no.6
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    • pp.353-358
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    • 2023
  • In the present study, to determine the efficacy of oral supplementation of ginseng berry extracts in augmenting exercise performance and exercise-associated metabolism, male mice were given orally 200 and 400 mg/kg of body weight (BW) of GBC for nine weeks. Although there are no differences in pre-exercise blood lactate levels among (1) the control group that received neither exercise nor GBC, (2) the group that performed only twice-weekly endurance exercise, and (3) and (4) the groups that combined twice-weekly endurance exercise with either 200 or 400 mg/kg GBC, statistically significant reductions in post-exercise blood lactate levels were observed in the groups that combined twice-weekly endurance exercise with oral administration of either 200 or 400 mg/kg GBC. Histological analysis showed no muscle hypertrophy, but transcriptome analysis revealed changes in gene sets related to lactate metabolism and mitochondrial function. GBC intake increased nicotinamide adenine dinucleotide levels in the gastrocnemius, possibly enhancing the mitochondrial electron transport system and lactate metabolism. Further molecular mechanisms are needed to confirm this hypothesis.

4-Hydroxynonenal Promotes Growth and Angiogenesis of Breast Cancer Cells through HIF-1α Stabilization

  • Li, Yao-Ping;Tian, Fu-Guo;Shi, Peng-Cheng;Guo, Ling-Yun;Wu, Hai-Ming;Chen, Run-Qi;Xue, Jin-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10151-10156
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    • 2015
  • 4-Hydroxynonenal (4-HNE) is a stable end product of lipid peroxidation, which has been shown to play an important role in cell signal transduction, while increasing cell growth and differentiation. 4-HNE could inhibit phosphatase and tensin homolog (PTEN) activity in hepatocytes and increased levels have been found in human invasive breast cancer. Here we report that 4-HNE increased the cell growth of breast cancer cells as revealed by colony formation assay. Moreover, vascular endothelial growth factor (VEGF) expression was elevated, while protein levels of hypoxia inducible factor 1 alpha (HIF-$1{\alpha}$) were up-regulated. Sirtuin-3 (SIRT3), a major mitochondria NAD+-dependent deacetylase, is reported to destabilize HIF-$1{\alpha}$. Here, 4-HNE could inhibit the deacetylase activity of SIRT3 by thiol-specific modification. We further demonstrated that the regulation by 4-HNE of levels of HIF-$1{\alpha}$ and VEGF depends on SIRT3. Consistent with this, 4-HNE could not increase the cell growth in SIRT3 knockdown breast cancer cells. Additionally, 4-HNE promoted angiogenesis and invasion of breast cancer cells in a SIRT3-dependent manner. In conclusion, we propose that 4-HNE promotes growth, invasion and angiogenesis of breast cancer cells through the SIRT3-HIF-$1{\alpha}$-VEGF axis.

Duodenal-Jejunal Bypass Surgery Stimulates the Expressions of Hepatic Sirtuin1 and 3 and Hypothalamic Sirtuin1

  • Ha, Eunyoung;Kang, Jong Yeon;Park, Kyung Sik;Seo, Youn Kyoung;Ha, Tae Kyung
    • Journal of Obesity & Metabolic Syndrome
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    • v.27 no.4
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    • pp.248-253
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    • 2018
  • Background: Sirtuins mediate metabolic responses to nutrient availability and slow aging and accompanying decline in health. This study was designed to assess the expressions of sirtuin1 (SIRT1) and sirtuin3 (SIRT3) in the liver and hypothalamus after duodenal-jejunal bypass (DJB) surgery in rats. Methods: A total of 38 rats were randomly assigned to either sham group (n=8) or DJB group (n=30). DJB group was again divided into three groups according to the elapsed time after surgery (10 weeks, DJB10; 16 week, DJB16; 28 week, DJB28). The mRNA and protein expressions of SIRT1 and SIRT3 in the liver and hypothalamus were measured by reverse transcription polymerase chain reaction, Western blot, and immunohistochemistry analyses. $NAD^+/NADH$ ratio was also measured. Results: We found increased mRNA and protein expression levels of SIRT1 in the liver of DJB16 and DJB28 groups compared with those of sham group. The mRNA and protein expressions of SIRT3 in the liver of DJB group increased proportionally to the elapsed time after DJB surgery. The mRNA expression levels of SIRT1 in the hypothalamus increased in DJB16 and DJB28 groups and protein expression levels of SIRT1 in the hypothalamus increased in DJB10, DBJ16, and DJB28 groups compared with sham group. We observed that mRNA and protein levels of SIRT3 in the hypothalamus of DJB group were not changed. Conclusion: This study proves that DJB increases SIRT1 and SIRT3 expressions in the liver and SIRT1 expression in the hypothalamus. These results suggest the possibility of sirtuins being involved in bypass surgery-induced metabolic changes.

Characteristics of Metallic Elements Concentration of Fine Particles(PM10, PM2.5) at Busan in 2004 (2004년 부산지역 미세먼지(PM10, PM2.5) 중의 금속 농도 특성)

  • Jeon Byung-Il;Hwang Yong-Sik;Lee Hyeok-Woo;Yang Ah-Reum;Kim Hyun-Jung;Seol Jae-Hwan;Kang Young-Jin;Kim Taek-Hoon;Jang Hyun-Seok
    • Journal of the Korean earth science society
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    • v.26 no.6
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    • pp.573-583
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    • 2005
  • [ $PM_{10}\;and\;PM_{2.5}$ ] aerosols were collected at Busan from March, 2004 to December, 2004, and the concentrations of some metal elements were chemically analyzed to study their characteristics. The mean concentration of $PM_{10}$ was $58.2{\mu}g/m^3$ with a range of 8.3 to $161.1{\mu}g/m^3$. The mean concentration of $PM_{2.5}$ was $29.3{\mu}g/m^3$ with a range of 2.8 to $65.3\mu}g/m^3$. The mean mass concentrations of Asian dust and non Asian dust in $PM_{10}$ were $121.5\mu}g/m^3$ and $56.0{\mu}g/,^3$ respectively. The mean values of crustal enrichment factors for six elements (Cd, Cr, Cu, Ni, Pb and Zn) were all higher than 10, possibly suggesting the influence of anthropogenic sources. The crustal enrichment factors of some heavy metal elements in non-Asian dust (NAD) were higher than those in Asian dust (AD), possibly due to anthropogenic emissions transported from industries around this area by westerly wind. The soil contribution ratios for $PM_{10}$ and $PM_{2.5}$ were $15.2\%$ and $17.5\%$ on the whole. and those of AD/NAD for $PM_{10}$ and $PM_{2.5}$ were 1.9 and 2.1, respectively.

Synergistic Efficacy of Concurrent Treatment with Cilostazol and Probucol on the Suppression of Reactive Oxygen Species and Inflammatory Markers in Cultured Human Coronary Artery Endothelial Cells

  • Park, So-Youn;Lee, Jeong-Hyun;Shin, Hwa-Kyoung;Kim, Chi-Dae;Lee, Won-Suk;Rhim, Byung-Yong;Shin, Yung-Woo;Hong, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.4
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    • pp.165-170
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    • 2008
  • In the present study, we aimed to identify the synergistic effects of concurrent treatment of low concentrations of cilostazol and probucol to inhibit the oxidative stress with suppression of inflammatory markers in the cultured human coronary artery endothelial cells (HCAECs). Combination of cilostazol (0.3${\sim}3{\mu}$M) with probucol (0.03${\sim}0.3{\mu}$M) significantly suppressed TNF-${\alpha}$-stimulated NAD(P)H-dependent superoxide, lipopolysaccharide (LPS)-induced intracellular reactive oxygen species (ROS) production and TNF-${\alpha}$ release in comparison with probucol or cilostazol alone. The combination of cilostazol (0.3${\sim}3{\mu}$M) with probucol (0.1${\sim}0.3{\mu}$M) inhibited the expression of vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemoattractant protein-1 (MCP-1) more significantly than did the monotherapy with either probucol or cilostazol. In line with these results, combination therapy significantly suppressed monocyte adhesion to endothelial cells. Taken together, it is suggested that the synergistic effectiveness of the combination therapy with cilostazol and probucol may provide a beneficial therapeutic window in preventing atherosclerosis and protecting from cerebral ischemic injury.