• Title/Summary/Keyword: cold shock protein

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Evaluation of Antifreeze Proteins on Miniature Pig Sperm Viability, DNA Damage, and Acrosome Status during Cryopreservation

  • Kim, Daeyoung
    • Journal of Embryo Transfer
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    • v.31 no.4
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    • pp.355-365
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    • 2016
  • The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

Ginsenosides Rg1 regulate lipid metabolism and temperature adaptation in Caenorhabditis elegans

  • Hao Shi ;Jiamin Zhao ;Yiwen Li ;Junjie Li ;Yunjia Li;Jia Zhang ;Zhantu Qiu ;Chaofeng Wu ;Mengchen Qin ;Chang Liu ;Zhiyun Zeng ;Chao Zhang ;Lei Gao
    • Journal of Ginseng Research
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    • v.47 no.4
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    • pp.524-533
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    • 2023
  • Background: Obesity is a risk factor for aging and many diseases, and the disorder of lipid metabolism makes it prominent. This study aims to investigate the effect of ginsenoside Rg1 on aging, lipid metabolism and stress resistance Methods: Rg1 was administered to Caenorhabditis elegans (C. elegans) cultured in NGM or GNGM. The lifespan, locomotory activity, lipid accumulation, cold and heat stress resistance and related mRNA expression of the worms were examined. Gene knockout mutants were used to clarify the effect on lipid metabolism of Rg1. GFP-binding mutants were used to observe the changes in protein expression Results: We reported that Rg1 reduced lipid accumulation and improved stress resistance in C. elegans. Rg1 significantly reduced the expression of fatty acid synthesis-related genes and lipid metabolism-related genes in C. elegans. However, Rg1 did not affect the fat storage in fat-5/fat-6 double mutant or nhr-49 mutant. Combined with network pharmacology, we clarified the possible pathways and targets of Rg1 in lipid metabolism. In addition, Rg1-treated C. elegans showed a higher expression of anti-oxidative genes and heat shock proteins, which might contribute to stress resistance Conclusion: Rg1 reduced fat accumulation by regulating lipid metabolism via nhr-49 and enhanced stress resistance by its antioxidant effect in C. elegans.

Effect of Suboptimal Temperature Incubation on the Resistance of Lactobacillus acidophilus CT 01 to Storage and Drying (저온배양에 따른 Lactobacillus acidophilus CT 01의 저장 및 건조에 대한 저항성)

  • Yu Keun-Hyung;Kwon Il-Kyoung;Kim Gur-Yoo
    • Food Science of Animal Resources
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    • v.25 no.1
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    • pp.92-97
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    • 2005
  • This study was carried out to determine the storage, cryotolerance, heat and drying resistance, when Lactobacillus acidophilus CT 01 isolated from preweaned piglet feces growing at suboptimal temperature. L. acidophilus CT 01 suboptimal temperature incubated for 48 hours had the slowest growth rate at 22℃ but the highest viable cell number after 36 hours at 22℃, with 1.3×10/sup 9/ CFU/mL. In case of 4 and 20℃ storage, the suboptimal temperature incubated groups had a viability higher than the control (p<0.01). The cryotolerance of suboptimal temperature incubated L. acidophilus CT 01 was a higher than the control (p<0.01). When L. acidophilus CT 01 was heat treated at 60℃ for 15 minutes and 30 minutes, the suboptimal temperature incubated L. acidophilus CT 01 at 22℃ had a viability higher more than the control (p<0.01). L. acidophilus CT 01 incubated suboptimal temperature was inoculated by 30% to the carrier, and dried at 50℃ for 12 hours had the highest viability in the suboptimal temperature incubated L. acidophilus CT 01 at 28℃.

The Effect of Linarin on LPS-Induced Cytokine Production and Nitric Oxide Inhibition in Murine Macrophages Cell Line RAW264.7

  • Kim, Kyung-Jae;Han, Shin-Ha;Sung, Ki-Hyun;Yim, Dong-Sool;Lee, Sook-Kyeon;Lee, Chong-Kil;Ha, Nam-Ju
    • Archives of Pharmacal Research
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    • v.25 no.2
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    • pp.170-177
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    • 2002
  • The herb, Chrysanthemum zawadskii var, latilobum commonly known as Gu-Jul-Cho in Korea, used in traditional medicine to treat pneumonia, bronchitis, cough, common cold, pharyngitis, bladder-related disorders, gastroenteric disorders, and hypertension. Linarin is the main active compound and the biological mechanisms of its activity are unclear. It is believed that effects of this herb may be exerted through the pluripotent effectors of linarin due to its ability to treat a variety of afflictions. In this study, the effects of linarin on the mouse macrophages cell line, RAW 264.7, were investigated. It was found that linarin could activate macrophages by producing cytokines. Monocytes and tissue macrophages produce at least two groups of protein mediators of inflammation, interleukin 1 (IL-1 ) and the tumor necrosis factor (TNF). Recent studies have shown that TNF and IL-1 modulate the inflammatory function of endothelial cells, leukocytes, and fibroblasts. $TNF-{\alpha}$ production by macrophages treated with linarin occured in a dose dependent manner However, IL-1 production was largely unaffected by this natural product. This study demonstrated the ability of linarin to activate macrophages both directly and indirectly. Linarin also affect both cytosine production and nitric oxide inhibition, in addition to the expression of some surface molecules. Nitric oxide (NO), derived from L-argin-ine, is produced by two forms(constitutive and inducible) of nitric oxide synthase (NOS). The NO produced in large amounts by inducible NOS is known to be responsible for the vasodilation and hypotension observed in septic shock. Linarin was found to inhibit NO production in the LPS-activated RAW 264.7 cells. Linarin may be a useful candidate as a new drug for treating endotoxemia and the inflammation accompanied by NO overproduction. The linarin-treated total Iymphocytes exhibited cytotoxicity in a dose dependent manner between $20{\;}{\mu}g/ml{\;}and{\;}40{\;}{\mu}g/ml$. These results suggest that linarin may function through macrophage activation.

Accelerated Growth of Corynebacterium glutamicum by Up-Regulating Stress-Responsive Genes Based on Transcriptome Analysis of a Fast-Doubling Evolved Strain

  • Park, Jihoon;Lee, SuRin;Lee, Min Ju;Park, Kyunghoon;Lee, Seungki;Kim, Jihyun F.;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1420-1429
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    • 2020
  • Corynebacterium glutamicum, an important industrial strain, has a relatively slower reproduction rate. To acquire a growth-boosted C. glutamicum, a descendant strain was isolated from a continuous culture after 600 generations. The isolated descendant C. glutamicum, JH41 strain, was able to double 58% faster (td=1.15 h) than the parental type strain (PT, td=1.82 h). To understand the factors boosting reproduction, the transcriptomes of JH41 and PT strains were compared. The mRNAs involved in respiration and TCA cycle were upregulated. The intracellular ATP of the JH41 strain was 50% greater than the PT strain. The upregulation of NCgl1610 operon (a putative dyp-type heme peroxidase, a putative copper chaperone, and a putative copper importer) that presumed to role in the assembly and redox control of cytochrome c oxidase was found in the JH41 transcriptome. Plasmid-driven expression of the operon enabled the PT strain to double 19% faster (td=1.82 h) than its control (td=2.17 h) with 14% greater activity of cytochrome c oxidase and 27% greater intracellular ATP under the oxidative stress conditions. Upregulations of genes those might enhance translation fitness were also found in the JH41 transcriptome. Plasmid-driven expressions of NCgl0171 (encoding a cold-shock protein) and NCgl2435 (encoding a putative peptidyl-tRNA hydrolase) enabled the PT to double 22% and 32% faster than its control, respectively (empty vector: td=1.93 h, CspA: td=1.58 h, and Pth: td=1.44 h). Based on the results, the factors boosting growth rate in C. gluctamicum were further discussed in the viewpoints of cellular energy state, oxidative stress management, and translation.