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Alpha-linolenic acid enhances maturation and developmental competence via regulation of glutathione, cAMP and fatty acid accumulation during in vitro maturation of porcine oocytes

  • Jeon, Ye-Eun;Hwangbo, Yong;Kim, Sun-Young;Park, Choon-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.357-365
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    • 2020
  • The aim of present study was to investigate regulatory mechanism of alpha-linolenic acid (ALA) during in vitro maturation (IVM) on nuclear and cytoplasmic maturation of porcine oocytes. Basically, immature cumulus-oocyte complexes (COCs) were incubated for 22 h in IVM-I to which hormone was added, and then further incubated for 22 h in IVM-II without hormone. As a result, relative cumulus expansion was increased at 22 h after IVM and it was enhanced by treatment of ALA compared with control group (p < 0.05). During IVM process within 22 h, cAMP level in oocytes was decreased at 6 h (p < 0.05) and it was recovered at 12 h in ALA-treated group, while oocytes in control group recovered cAMP level at 22 h. In cumulus cells, it was reduced in all time point (p < 0.05) and ALA did not affect. Treatment of ALA enhanced metaphase-I (MI) and MII population of oocytes compared with oocytes in control group at 22 and 44 h, respectively (p < 0.05). Intracellular GSH levels in ALA group was increased at 22 and 44 h after IVM (p < 0.05), whereas it was increased in control group at 44 h after IVM (p < 0.05). In particular, the GSH in ALA-treated oocytes during 22 h of IVM was higher than control group at 22 h (p < 0.05). Lipid amount in oocytes from ALA group was higher than control group (p < 0.05). Treatment of ALA did not influence to absorption of glucose from medium. Cleavage and blastocyst formation of ALA-treated oocytes were enhanced compared with control group (p < 0.05). These findings suggest that supplementation of ALA could improve oocyte maturation and development competence through increasing GSH synthesis, lipid storage, and regulation of cAMP accumulation during early 22 h of IVM, and these might be mediated by cumulus expansion.

Effects of Roscovitine on In Vitro Development of Porcine Oocyte Using Brilliant Cresyl Blue

  • Roy, Pantu Kumar;Fang, Xun;Hassan, Bahia MS;Shin, Sang Tae;Cho, Jong Ki
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.111-122
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    • 2017
  • The objective of this experiment was to explore the effects of Roscovitine (Rosco) prior to in vitro maturation (IVM) of immature pig oocyte. Brilliant cresyl blue test has been used to select the good quality of oocyte. Specifically, the effects of Rosco exposure on nuclear and cytoplasmic maturation, diameter, intracellular glutathione (GSH) and reactive oxygen species (ROS), and embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT), and gene expression levels in SCNT embryos have been measured. Cumulus oocyte complexes (COCs) have been exposed in $75{\mu}M$ of Rosco for 22 and 44 h. The COCs that were matured in the IVM for 44 h without Rosco used as control group. Diameter of matured porcine oocytes 44 h culture with Rosco was significantly lower than 22 h culture with Rosco and control groups. GSH was higher in control group than 22 h and 44 h with Rosco but reduction of ROS in 22 h than 44 h with Rosco. In PA, exposure with Rosco 44 h oocytes group has been significantly lower than 22 h and control group in rates of maturation, cleavage and blastocyst formation. Similarly, in SCNT embryos rates of maturation, cleavage and formation of blastocyst have been also significantly lower in 44 h Rosco treated group than other two groups. SCNT embryos treated with Rosco 22 h showed greater expression levels of POU5F1, DPPA2 and NDP52Il mRNA compared with other two groups. Our results demonstrate that Rosco treatment with 22 h prior to IVM improves the development competence of porcine oocyte.

Effects of Photo/dark period and Relative Humidity during Dark Period on Growth and Tipburn Occurrence of Water Dropwort (Oenanthe stolonifera DC.) in a Closed-type Plant Factory (밀폐형 식물공장에서 명/암주기와 암기동안의 상대습도가 미나리 생육과 팁번 발생에 미치는 영향)

  • An, Jae Uk;Joung, Kyoung Hee;Yoon, Hae Suk;Hwang, Yeon Hyeon;Hong, Gwang Pyo
    • Journal of Bio-Environment Control
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    • v.26 no.2
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    • pp.146-150
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    • 2017
  • This research investigated the effect of photo/dark period and relative humidity during dark period on the growth and quality of water dropwort in a closed-type plant factory system. At 30 days after planting, the shoot fresh weight of water dropwort under relative humidity of 60/90%(light/dark) treatment significantly higher than that under relative humidity of 60/60% treatment. The shoot fresh weight of water dropwort increased by extending light period under relative humidity of 60/60% treatment, but 16/8h photo/dark period showed the best shoot fresh weight, followed by 20/4h and 22/2h under relative humidity of 60/90% treatment. In the relative humidity of 60/90% treatment, the tipburn occurrence was reduced under 16/8h photo/dark period condition as 1.4%, whereas it was significantly increased under 20/4h and 22/2h of relatively long light time duration as 15.5% and 30.3%, respectively. In the relative humidity of 60/60% treatment, the tipburn occurrence was 15.5% under 16/8h photo/dark period condition and those under 20/4h and 22/2h photo/dark period condition were higher than 25%. The stem hardness of water dropwort was lowest in relative humidity of 60/90% and 16/8h photo/dark period treatment. The mineral contents of leaves were decreased by extending light period, but the contents of Ca were not different significantly among the treatments except the 60/60% and 22/2h treatment.

Propylene Epoxidation Using Ti-MCM-22 Catalyst (Ti-MCM-22 촉매를 이용한 프로필렌 에폭시화반응)

  • Yang, Seung-Tae;Ban, Han-Ju;Kim, Se-Young;Ahn, Wha-Seung
    • Korean Chemical Engineering Research
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    • v.46 no.4
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    • pp.665-668
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    • 2008
  • Propylene epoxidation by $H_2O_2$ (30% aqueous) as oxidant was studied in a semi-batch reactor using Ti-MCM-22 catalyst: Effects of reaction temperature, pressure, catalyst loading, solvent, and $H_2O_2$ concentration on $H_2O_2$ conversion (limiting reagent) were investigated. Product inhibition by propylene oxide was confirmed. Ti-MCM-22 maintained virtually the same catalytic performance over the 5 repeated cycles.

Neuroprotective Effect of Rice with Phellinus linteus Mycelium in HT22 Cells (상황버섯균사체 쌀의 HT22 신경세포 보호 효과)

  • Kim, Ji Hyun;Chun, Soon Sil
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.7
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    • pp.886-890
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    • 2017
  • In this study, the protective effect of rice with Phellinus linteus mycelium (PLMR) against hydrogen peroxide-induced oxidative stress was assessed in a mouse hippocampal neuronal HT22 cell line through (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) salt (MTS) assay and western blot. MTS assay using HT22 cells showed that PLMR extract did not affect viability at a concentration range from 1 mg/mL to 5 mg/mL. However, at concentrations over 10 mg/mL, PLMR extract resulted in increased cell death. Cell viability of HT22 was significantly reduced by $H_2O_2$ treatment, and reduction of cell viability was efficiently restored by treatment with PLMR extract in a dose-dependent manner from 0.1 to 1 mg/mL. Cells treated with $H_2O_2$ showed increased expression of Bax, a pro-apoptotic protein, which was down-regulated by treatment with PLMR extract. On the other hand, cells treated with $H_2O_2$ resulted in reduced expression of Bcl-2, an anti-apoptotic protein, which was restored by treatment with PLMR extract. In addition, treatment with PLMR extract reduced expression of cleaved caspase 3 and PARP, which were up-regulated by $H_2O_2$ treatment. The results may suggest that treatment with PLMR extract would suppress $H_2O_2$-induced apoptosis of HT22 cells.

Analysis of Technology Convergence of 'Internet of Things' Patents by IPC Code Analysis (IPC 코드 분석에 의한 '사물인터넷(IoT)' 특허의 기술 융복합 분석)

  • Shim, Jae-ruen
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.9 no.3
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    • pp.266-272
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    • 2016
  • In this study, we analysed the technology convergence of Internet of Things(IoT) by International Patent Classification(IPC) code analysis. 163 patent applications in Korea are the subjects of this study. We confirmed that the most representative IPC code combinations between Main and Sub categories are G06Q 50/24-G06Q 50/22(6 cases), H04L 29/02-H04L 12/28(4 cases), G06F 15/16-G06F 3/048(3 cases), G06F 15/16-G06F 9/44(3 cases), and G06Q 50/22-G06Q 50/24(3 cases). The field of Health Care business have been prepared 9 patent applications by technology convergence between 'Health Care(G06Q 50/22)' and 'Patient Record Management(G06Q 50/24)'. Finally we also concluded that the core IPC code are G06F 15/16, G06Q 50/22, G06Q 50/24, and H04L 12/28 by the technology convergence interconnections analysis of IoT patent applications.

Effects of $H_2O_2$ and ascorbic acid on TIMP-2, Type1 collagen, and PDLs22 levels in human periodontal ligament fibroblasts (($H_2O_2$와 ascorbic acid가 사람 치주인대섬유모세포의 TIMP-2, Type 1 collagen, PDLs22 발현에 끼치는 효과)

  • Choi, Yong-Sun;Kim, So-Young;Choi, Seong-Mi;Jang, Hyun-Seon;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.37 no.4
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    • pp.655-669
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    • 2007
  • Reactive oxygen species (ROS) have been implicated in the pathogenesis of various diseases. And vitamin C has shown a protective effect for the tissues. The aim of this study was to evaluate the effects of $H_2O_2$ and ascorbic acid on matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase (TIMP: TIMP-1, TIMP-2), Type 1 collagen, fibronectin, and PDLs22 level in human periodontal ligament fibroblasts (hPDLF) via reverse transcription-polymerase chain reaction (RT-PCR). hPDLF was obtained from a healthy periodontium and cultured in Dulbecco's modified Eagles's medium plus 10% fetal bone serum. The concentration of ascorbic acid in hPDLF was $50{\mu}g/ml$, and that of $H_2O_2$ in hPDLF was 0.03% and 0.00003%. Ascorbic acid only, $H_2O_2$ only and mixture of ascorbic acid and $H_2O_2$ were applied with hPDLF for 1-, 3-, and 30-min. respectively. The gene expression of MMP-1-, TIMP-1-, TIMP-2-, Type 1 collagen-, fibronectin-, and PDLs22-mRNA in hPDLF was analysed via RT-PCR. The results were as follows; 1. hPDLF in response to 30-min. incubation with 0.03% $H_2O_2$ did not show any gene expression. 2. In all the experimental groups, the gene expression of fibronectin mRNA showed the decreased tendency compared to control. 3. In all the experimental groups, the gene expression of TIMP-1 mRNA showed the tendency similar to control. 4. hPDLF in response to 30-min. incubation with 0.03% $H_2O_2$ and ascorbic acid increased mRNA induction for MMP-1. 5. In all the experimental groups, hPDLF increased mRNA induction for PDLs22, collagen type 1, and TIMP-2 compared to control. Within the limited experiments, $H_2O_2$ and ascorbic acid increased mRNA induction for PDLs22, collagen type 1, TIMP-2 in hPDLF. More research will be needed in order to confirm the relative importance of the different roles of ROS and antioxidants in hPDLF from a periodontal regeneration or repair standpoint.

Effect of 0.5 mM Dibutyryl cAMP on Meiotic Maturation during Different Incubation Time and Embryonic Development Following In Vitro Fertilization or Parthenogenetic Activation in Porcine Oocytes

  • Yu, Il-Jeoung
    • Journal of Embryo Transfer
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    • v.26 no.4
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    • pp.251-256
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    • 2011
  • Presently, the effect of 0.5 mM dibutyryl cAMP (dbcAMP)-supplemented maturation medium during different incubation time on meiotic arrest (germinal vesicle) and resumption (metaphase II) of porcine oocytes and embryonic development of porcine oocytes following in vitro fertilization (IVF) or parthenogenetic activation (PA) was determined. Porcine cumulus oocyte complexes (COCs) were cultured in 0.5 mM dbcAMP for 17, 22, 27, or 42 h, and an additional 22 h without 0.5 mM dbcAMP. The nuclear status was examined at each time point. Oocytes cultured from 39~49 h displayed more than 80% meiotic resumption. More than 85 % of meiotic arrest was presented at 17~22 h. Oocytes were cultured for 22 h with 0.5 mM dbcAMP and additional 22 h without dbcAMP to assess developmental potential following IVF or PA. There were no significant differences in blastocyst rates among the dbcAMPIVF, IVF, dbcAMP-PA, and PA groups, although cleavage rate of IVF group was significantly higher than those of dbcAMP-PA, and PA groups. In conclusion, 0.5 mM dbcAMP influenced meiotic maturation of porcine oocytes depending on incubation time of oocyte, although embryonic development was not improved in both IVF and PA.

Isolation of Bacteriocin-producing Lactic Acid Bacteria from Human Intestines and the Characteristics of their Bacteriocins (Bacteriocin을 생산하는 장내 유산균의 분리 및 Bacteriocin 특성조사)

  • 김정환;맹길재;김정상;지근억
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1228-1236
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    • 1997
  • Lactobacillus strains were isolated from volunteer's feces, including from newly-born infants and adults in their 20's, by using differential MRS-BPB plates. Total 56 presumptive Lactobacillus strains were isolated and the bacteriocin productions by the isolates were examined by agar diffusion method. Six bacteriocin-producing strains were confirmed. Among them, two isolates, HU-1 and H22-3, showed the most outstanding antimicrobial activities, which were not affected by pH adjustments or catalase treatments of culture. HU-1 was originated from a two-years old boy and H22-3 was originated from a newly-born infant. HU-1 and H22-3 had the same morphology(short rod) when examined by scanning electron microscope, and the same biochemical traits including growth temperature range, salt tolerance and sugar-fermenting abilities. But the growth-inhibition spectrum and plasmid profiles of HU-1 and H22-3 were different. Both strains inhibited the growth of various Gram (+) microorganisms including Listeria monocytogenes. Micrococcus luteus, and Staphylococcus aureus in addition to many species of lactic acid bacteria, indicating the production of broad-spectrum bacteriocins. Bacteriocins produced by HU-1 and H22-3 were stable up to 90℃, 15 min heat treatments. Their activities were not affected by pepsin or trypsin treatments but destroyed by proteinaseK or pronase treatments.

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