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Ginsenoside $R_e$ Increases Fertile and Asthenozoospermic Infertile Human Sperm Motility by Induction of Nitric Oxide Synthase

  • Zhang Hong;Zhou Qing-Ming;Li Xiao-Da;Xie Yi;Duan Xin;Min Feng-Ling;Liu Bing;Yuan Zhi-Gang
    • Archives of Pharmacal Research
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    • v.29 no.2
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    • pp.145-151
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    • 2006
  • We investigated the effects of Ginsenoside $R_e$ on human sperm motility in fertile and asthenozoospermic infertile individuals in vitro and the mechanism by which the Ginsenosides play their roles. The semen samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Spermatozoa were separated by Percoll and incubated with 0, 1, 10 or $100\;{\mu}M$ of Ginsenoside $R_e$. Total sperm motility and progressive motility were measured by computer-aided sperm analyzer (CASA). Nitric oxide synthase (NOS) activity was determined by the $^{3}H$-arginine to $^{3}H$-citrulline conversion assay, and the NOS protein was examined by the Western blot analysis. The production of sperm nitric oxide (NO) was detected using the Griess reaction. The results showed that Ginsenoside $R_e$ significantly enhanced both fertile and infertile sperm motility, NOS activity and NO production in a concentration-dependent manner. Sodium nitroprusside (SNP, 100 nM), a NO donor, mimicked the effects of Ginsenoside $R_e$. And pretreatment with a NOS inhibitor $N^{w}$-Nitro-L-arginine methyl ester (L-NAME, $100\;{\mu}M$) or a NO scavenger N-Acetyl-L-cysteine (LNAC, 1 mM) completely blocked the effects of Ginsenoside $R_e$. Data suggested that Ginsenoside $R_e$ is beneficial to sperm motility, and that induction of NOS to increase NO production may be involved in this benefit.

Effects of dry period length on milk production and physiological responses of heat-stressed dairy cows during the transition period

  • Dong-Hyun Lim;Da Jin Sol Jung;Kwang-Seok Ki;Dong-Hyeon Kim;Manhye Han;Younghoon Kim
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.197-208
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    • 2023
  • The objective of this study was to investigate the effects of a traditional dry period (60 d) versus a no dry period (0 d) on the milk production, physiological response, and metabolic status of dairy cows exposed to heat stress during the transition period. Holstein dairy cows (n = 15) with similar expected calving dates were randomly assigned to two different dry period lengths: (1) no dry period (n = 7) and (2) a traditional dry period of 60 days (n = 8). All cows were studied from 8 weeks before expected calving to 10 weeks after calving and experienced heat stress during the transition period. The results showed that cows with no dry period decreased their milk yield in subsequent lactation, but compensated for the loss of milk yield accounted for by additional milk yield before calving. The energy balance at postpartum was improved in cows with no dry period compared to cows with a traditional dry period. There were no significant differences in the physiological response and blood metabolites at postpartum between the dry period lengths of dairy cows exposed to heat stress during the transition period. Taken together, our results showed that omitting the dry period improved the milk production and metabolic status of dairy cows exposed to heat stress during the transition period.

Anti-Inflammatory and Antioxidant Effect of Astaxanthin Derived from Microalgae (미세조류 유래 astaxanthin의 항염증 및 항산화 효과)

  • Kwak, Tae-Won;Cha, Ji-Young;Lee, Chul-Won;Kim, Young-Min;Yoo, Byung-Hong;Kim, Sung-Gu;Kim, Jong-Myoung;Park, Seong-Ha;An, Won-Gun
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1377-1384
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    • 2011
  • Astaxanthin (ATX) is a red-orange carotenoid pigment that occurs naturally in a wide variety of living organisms. In this study we investigated the inhibitory effects of ATX on the induction of inducible nitric oxide synthase (iNOS), nitric oxide (NO), proinflammatory cytokines, nuclear factor-kappa B(NF-${\kappa}B$) and reactive oxygen species (ROS) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In addition, we tested the superoxide radical scavenging activity of ATX by scavenging assay. iNOS and NF-${\kappa}B$ expressions were determined by immunoblot analysis. Interleukin (IL)-6 and tumour necrosis factor-${\alpha}$ (TNF-${\alpha}$) were assayed by ELISA. NO production was monitored by measuring the amount of nitrite. ROS was examined by using the 2', 7'-Dichlorodihydrofluorescin diacetate (DCFH-DA) method. At a concentration of 100 ${\mu}M$, ATX inhibited the expression level of LPS-induced NF-${\kappa}B$, as well as the production of LPS-induced NO and proinflammatory cytokines (IL-6 and TNF-${\alpha}$), by suppressing iNOS expression. In particular, the maximal inhibition rate of IL-6 and TNF-${\alpha}$ production by ATX (100 ${\mu}M$) was 65.2----- and 21.2-----, respectively. In addition, ATX inhibited the LPS-induced transcriptional activity of NF-${\kappa}B$, and this was associated with suppressing the translocations of NF-${\kappa}B$ from the cytosol to the nucleus. Moreover, at various concentrations (25-100 ${\mu}M$), ATX inhibited the intracellular level of ROS. At a concentration of 5 mg/ml, the superoxide radical scavenging activity of ATX was 1.33 times higher than ${\alpha}$-tocopherol of the same concentration. These results showed that ATX inhibited the expression of iNOS and the production of NO and proinflammatory cytokines resulting from ROS production and NF-${\kappa}B$ activation in macrophages. Furthermore, ATX was found to be more effective in superoxide radical scavenging activities compared to ${\alpha}$-tocopherol. These findings are expected to strengthen the position of ATX as anti-inflammatory medicine and antioxidant.

Structure and properties of silk sericin obtained from different silkworm varieties

  • Chung, Da Eun;Lee, Ji Hye;Kweon, HaeYong;Lee, Kwang-Gill;Um, In Chul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.30 no.2
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    • pp.81-85
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    • 2015
  • Recently, many researchers have studied silk sericin because of its high water retention, good wound healing, good cyto-compatibility, and blood-glucose- and cholesterol-lowering effects. Although sericin film can be used in wound dressing and cosmetic packs, its poor mechanical properties have prevented its use in industrial fields. In the present study, sericin was obtained from different silkworm varieties, and the effect of silkworm variety on the structure and properties of sericin was examined. Except for a small difference in serine content, no significant difference in sericin was noted among the silkworm varieties. In addition, silkworm variety almost had no effect on solution viscosity, implying that it does not influence the molecular weight of sericin. Mechanical properties of sericin film were strongly affected by silkworm variety. Wonwon 126 showed the best mechanical properties, while N74 and Geumgwangju displayed the worst properties.

The Characteristics of the Flow and Combustion in a Turbulent Non-Premixed Flat Flame (난류 비예혼합 평면화염의 유동과 연소 특성)

  • Kwark, Ji-Hyun;Jung, Yong-Ki;Jun, Chung-Hwan;Chang, Young-June
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.27 no.4
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    • pp.447-457
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    • 2003
  • An experiment in a turbulent non-premixed flat flame was carried out in order to investigate the effect of swirl number on the flow and combustion characteristics. First. stream lines and velocity distribution in the flow field were obtained using PIV method. In contrast with the axial flow without swirl, highly swirled air induced stream lines along the burner tile. and backward flow was caused by recirculation in the center zone of the flow field. In the combustion. the flame with swirled air also became flat and stable along the burner tile with increment of the swirl number. Flame structure by measuring OH and CH radicals intensity and by calculating Damkohler number(Da) and turbulence Reynolds number(Re$_{T}$) was examined. It appeared to be comprised in the wrinkled laminar-flame regime. Backward flow by recirculation of the burned gas decreased the flame temperature and emissions concentrations as NO and CO. Consequently, the stable flat flame with low NO concentration was achieved.d.

Screening and Purification of an Antimicrobial Peptide from the Gill of the Manila Clam Ruditapes philippinarum (바지락(Ruditapes philippinarum) 아가미로부터 항균 펩타이드의 탐색 및 정제)

  • Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.2
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    • pp.137-145
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    • 2016
  • This study screened the biological activity of an acidified gill extract of the Manila clam Ruditapes philippinarum including antimicrobial, hemolytic, membrane permeabilization, and DNA-binding activity, and purified the antimicrobial material. The acidified gill extract showed potent antimicrobial activity against Bacillus subtilis and Escherichia coli without significant hemolytic activity, but showed no membrane permeabilization or DNA-binding ability. An antimicrobial material was purified from the acidified gill extract using C18 reversed-phase and cation-exchange high-performance liquid chromatography (HPLC). Treatment of the purified material with trypsin completely abolished all of the antibacterial activity against Bacillus subtilis, suggesting that the purified material is a proteinaceous antibiotic. The molecular weight of the purified material was 2571.9 Da, but no primary structural information was obtained due to N-terminal blocking. A future study should confirm the primary structure. Our results suggest that the Manila clam gill contains proteinaceous antibiotics that have a role in first-line defense. This information could be used to better understand the Manila clam innate immune system.

Overexpression of Adenosine Deaminase in Escherichia coli

  • Jo, Young-Bae;Baik, Hyung-Suk;Bae, Kyung-Mi;Jun, Hong-Ki
    • Journal of Life Science
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    • v.9 no.2
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    • pp.62-66
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    • 1999
  • To overexpress E. coli ADA in host strain E. coli M15, the expression plasmid pQEADD was constructed. To analyze the expression characteristics of ADA, a time course of the expression was first examined. The protein was not detected in no inducted in no induction samples. After the addition of IPTG, a band corresponding to the expected size of ADA was appeared. Its molecular weight was about 36,000 dalton. Maximum expression level was revealed when the cell cultured for 3∼4hrs after induction. This result indicated that the efficient expression of add can be achieved by induction at early logarithmic phase. The effect of different IPTG concentration on the degree of ADA expression was investigated. The expression levels of add were not largely affected by IPTG concentration. Location of overexpression ADA was checked out. A protein band corresponding to the ADA was seen in only crude extract B(insoluble protein). This result suggests that ADA is in E. coli M15. The molecular weight of ADA estimated by SDS-PAGE was approximately 36,000 Da.

Purification and Characterization of a Keratinase from a Feather-Degrading Fungus, Aspergillus flavus Strain K-03

  • Kim, Jeong-Dong
    • Mycobiology
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    • v.35 no.4
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    • pp.219-225
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    • 2007
  • A keratinolytic enzyme secreted by Aspergillus flavus K-03 cultured in feather meal basal medium (FMBM) containing 2% (w/v) chicken feather was purified and characterized. Keratinolytic enzyme secretion was the maximal at day 16 of the incubation period at pH 8 and $28^{\circ}C$. No relationship was detected between enzyme yield and increase of fungal biomass. The fraction obtained at 80% ammonium sulfate saturation showed 2.39-fold purification and was further purified by gel filtration in Sephadex G-100 followed by ion exchange chromatography on DEAE-Sephadex A-50, yielding an active protein peak showing 11.53-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymograms indicated that the purified keratinase is a monomeric enzyme with 31 kDa molecular weight. The extracellular keratinase of A. flavus was active in a board range of pH ($7{\sim}10$) and temperature ($30^{\circ}C{\sim}70^{\circ}C$) profiles with the optimal for keratinase activity at pH 8 and $45^{\circ}C$. The keratinase activity was totally inhibited by protease inhibitors such as phenylmethylsulfonyl fluoride (PMSF), iodoacetic acid, and ethylenediaminetetraacetate (EDTA) while no reduction of activity by the addition of dithiothreitol (DTT) was observed. N-terminal amino acid sequences were up to 80% homologous with the fungal subtilisins produced by Fusarium culmorum. Therefore, on the basis of these characteristics, the keratinase of A. flavus K-03 is determined to be subtilisins-like.

Anti-Inflammatory Activity of Ethanolic Extract of Sargassum micracanthum

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Ahn, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1691-1698
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    • 2013
  • The anti-inflammatory effects of Sargassum micracanthum ethanol extract (SMEE) was investigated using LPS-induced inflammatory response in this study. As a result, there was no cytotoxicity in the macrophage proliferation treated with SMEE compared with the control. SMEE inhibited production of nitric oxide and cytokines (IL-6, TNF-${\alpha}$, and IL-$1{\beta}$) in a dose-dependent manner. In addition, the expression of inducible nitric oxide synthase and cyclooxygenase 2 were suppressed via inhibition of nuclear factor ${\kappa}B$ p65 expression by SMEE treatment. The formation of edema in the mouse ear was reduced at the highest dose tested compared with that in the control, and reduction of ear thickness was observed in histological analysis. Moreover, in an acute toxicity test, no mortalities occurred in mice administered 5,000 mg/kg body weight of SMEE over a 2-week observation period. These results suggest that SMEE may have significant effects on inflammatory mediators and be a potential anti-inflammatory therapeutic material.

Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. -II. Purification of Trypsin from the Hepatopancreas of Penaeus orientalis-

  • Oh Eun-Sil;Kim Doo-Sang;Jung Kyoo-Jin;Pyeun Jae-Hyeung;Heu Min-Soo;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.1 no.2
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    • pp.209-215
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    • 1998
  • Trypsin-like enzyme was purified from shrimp hepatopancreas through Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Superdex 75 gel chromatography. Purity of trypsin-like enzyme was increased 69-fold with $44\%$ yield. The enzyme consisted of a single polypeptide chain with a molecular weight (M.W.) of 32 kDa judged by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was completely inactivated by serine enzyme inhibitors such as soybean trypsin inhibitor (SBTI), tosyl-L­lysine chloromethyl ketone (TLCK), and leupeptin. However, the enzyme was not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) which is a chymotrypsin specific inhibitor. The enzyme had no activity against benzoyl-tyrosine ethyl ester (BTEE) which is a chymotrypsin specific substrate. The enzyme showed high activity on the carboxyl terminal of Phe, Tyr. Glu, Arg, and Asp. However. no activity was detected against the carboxyl terminal of Pro, Trp, Cys, Gly, Val, and Ala.

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