• Title/Summary/Keyword: post-incubation

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Ultraviolet Radiation-Induced Apoptosis is Inversely Correlated with the Expression Level of Poly(ADP-ribose) Polymerase

  • Oh, Kyu Seon;Lee, Dong Wook;Chang, Jeong Hyun;Moon, Yong Suk;Um, Kyung ll
    • Animal cells and systems
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    • v.5 no.1
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    • pp.77-83
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    • 2001
  • The present study was conducted to elucidate whether the expression level of poly(ADP-ribose) polymerase (PARP) is related to the ultraviolet radiation (UV)-induced apoptosis. After treatment of the mammalian cell lines HeLa S3 and Chinese hamster ovary (CHO) with 50 J/m2 UV, induction of apoptosis was determined by several means during 24 h post-incubation. Incidence of apoptosis was much lower in CHO than HeLa S3 cells based on the percentage of apoptotic cells in terms of morphological changes in nucleus or direct counting of viable cells and qualitative or quantitative DNA fragmentation. Interestingly, when the expression level of PARP was measured by western blotting, the amounts of PARP that was retained at each time point inversely correlated with the incidences of apoptosis in these cells. Concomitant with generation of the 85 kDa fragment, 116 kDa PARP disappeared in HeLa S3 within 6 h after UV treatment, whereas a fair amounts of 116 kDa band was still retained in CHO cells at 36 h post-incubation. This inverse relationship was also observed in the adaptive response system, in which cells weve treated with a high dose of UV after pretreatment with a low dose. As expected, typical adaptive responses appeared in CHO cells but not in HeLa cells, showing greater cell viability and lesser DNA fragmentation. During the adaptive response in CHO cells, PARP was expressed at much higher level compared to the single, high dose-treated cells. Interestingly, even though PARP was induced at 6 h post-incubation In both cell types, its expression was more prominent in CHO cells. Thus, our data indicate that the retained level of intact PARP against UV damage inversely correlates with incidence of apoptosis in mammalian cells, and also suggest that a machinery to protect the PARP degradation against UV damage exists in CHO but not in HeLa S3 cells.

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Effect of Korean Red Ginseng Extract on Cell Death Responses in Peroxynitrite-Treated Keratinocytes

  • Kim, Hyoung-Do;Ha, Se-Eun;Kang, Jea-Ran;Park, Jong-Kun
    • Journal of Ginseng Research
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    • v.34 no.3
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    • pp.205-211
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    • 2010
  • Korean red ginseng (KRG) has been used worldwide as a traditional medicine for the treatment of various diseases, including cancer. In this study, we determined the effect of KRG on the responses of HaCaT cells to peroxynitrite ($ONOO^-$). Cells has been used worldwide as a traditional medicine for the treatment of various diseases, including cancer. In this study, we determined the effect of KRG on the responses of HaCaT cells to peroxynitrite ($ONOO^-$). Cells treated with $ONOO^-$ (2 mM) prior to incubation with control medium for 12 hours displayed reduced viability, as determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (viability about 48% of that of non-treated control cells). When KRG was added to the post-incubation medium, the negative effects of $ONOO^-$ on cell viability were significantly reduced. Reverse transcription-polymerase chain reaction analysis indicated that KRG alone did not significantly alter p53 or "growth arrest and DNA damage" (GADD)45 mRNA levels. However, the addition of KRG to the post-incubation medium significantly and dose-dependently reduced levels of p53 and GADD45 mRNA in $ONOO^-$-treated cells. Western blot analyses revealed that incubation with KRG decreased p53 and GADD45 protein levels in $ONOO^-$-treated cells, relative to those in cells incubated with control medium. Collectively, these results suggest that Korean red ginseng extract protects cells against $ONOO^-$-induced genotoxicity by increasing cell viability through modulating the expression of p53 signaling intermediates.

Effects of Incubation and Thawing Temperature on Frozen-thawed Stallion Epididymal Spermatozoa (말의 정소상체 정자의 동결 후 해동 온도 및 Incubation의 효과)

  • Kim, Keun-Jung;Lee, Kyung-Bon;Lee, Ji-Hye;Kim, Eun-Young;Han, Kil-Woo;Park, Kang-Sun;Kim, Min-Kyu
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.297-302
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    • 2013
  • Cryopreservation of epididymal spermatozoa offers a potential tool for rescuing genetic material from males of genetically elite populations. Castration, catastrophic injury, sudden death or any other event that makes semen collection or mating impossible may prematurely terminate a stallion reproduction. Stallion epididymal spermatozoa vary widely in the loss of progressive motility, acrosomal integrity, and viability during freezing and thawing. The objective of this work was to investigate the effect of (1) freezing package types on cryopreservation efficiency, (2) thawing temperatures (37, 56 or $70^{\circ}C$) on Computer Assisted Sperm Analysis (CASA) parameters and (3) post-thawing incubation time (0, 1, 2 or 4h) on castrated stallion epididymis. Post-thawed sperm motility ranged between 59.69% and 64.28% ($56^{\circ}C$ and $37^{\circ}C$) in various thawing temperatures. When stallion epididymis sperm was frozen, straw was better than freezing tube on VCL (Velocity of Curvilinear Line) and VAP (Velocity of Average Path) parameter. Higher percentage of motility was observed at $37^{\circ}C$ thawing temperature even though no significant difference was observed among various temperatures. The motility, VCL, ALH (Amplitude of Lateral Head displacement), VAP, BCF (Beat-Cross Frequency) and STR (Straightness index) parameter of post-thawed sperm were significantly decreased by increasing the incubation time for all thawing temperatures. The present study showed that type of freezing package (Straw vs. Freezing tube) was not significantly different on cryopreservation efficiency. Furthermore, stallion epididymal spermatozoa frozen-thawed at $37^{\circ}C$ for 1 min resulted the highest proportion of motility and velocity movement. In addition, motility and viability of frozen-thawed stallion epididymal spermatozoa were also decreased by incubation.

Effect of Fodder Tree Species with Condensed Tannin Contents on In vitro Methane Production

  • Vazquez, Ernestina Gutierrez;Medina, Leonardo Hernandez;Benavides, Liliana Marquez;Caratachea, Aureliano Juarez;Razo, Guillermo Salas;Burgos, Armin Javier Ayala;Rodriguez, Ruy Ortiz
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.1
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    • pp.73-79
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    • 2016
  • The objective was to evaluate the effect of fodder tree species (FTS) with condensed tannin contents: Cordia elaeagnoides, Platymiscium lasiocarpum, Vitex mollis, and Haematoxylon brasiletto, on in vitro methane ($CH_4$) production at 24 h post incubation. The analysis was performed using the in vitro gas production technique, with three levels of inclusion/species: 600, 800, and 1,000 mg and with 4 replicates/species/level of inclusion. The substrate was incubated at $39^{\circ}C$, and the gas and $CH_4$ production were recorded at 4, 8, 12, and 24 h post incubation. The data collected was analyzed through Pearson correlation, polinomial regression and fixed effects models. There were negative correlations between FTS-total gas volume (r = -0.40; p<0.001); FTS-volume of $CH_4$ produced (r = -0.40; p<0.001) and between the inclusion level-volume of $CH_4$ produced (r = -0.20; p<0.001). As well as a positive correlation between hours post incubation-total gas volume (r = 0.42; p<0.001) and between hours post incubation-volume of $CH_4$ produced (r = 0.48; p<0.001). The FTS: C. elaeagnoides, V. mollis, and H. brasiletto have potential, in the three inclusion levels analyzed, to reduce $CH_4$ emission on in vitro trials (>32.7%), taking into account the total $CH_4$ production at 24 h of the forage used as reference (Avena sativa). It's suggested that C. elaeagnoides-according to its crude protein, neutral detergent fiber, and condensed tannins content- is the best alternative within the FTS analyzed, for feeding ruminants and for the control of $CH_4$ emissions during the dry season.

The In Vitro and In Vivo Effect of Lipoxygenase Pathway Inhibitors Nordihydroguaiaretic Acid and Its Derivative Tetra-O-methyl Nordihydroguaiaretic Acid against Brucella abortus 544

  • Reyes, Alisha Wehdnesday Bernardo;Kim, Heejin;Huy, Tran Xuan Ngoc;Nguyen, Trang Thi;Min, Wongi;Lee, Dongho;Hur, Jin;Lee, John Hwa;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.32 no.9
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    • pp.1126-1133
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    • 2022
  • This study investigated the contribution of lipoxygenase (LOX) inhibitors, nordihydroguaiaretic acid (NDGA), tetra-O-methyl nordihydroguaiaretic acid (M4N) and zileuton (ZIL), and thromboxane A2 (TXA2) inhibitor 4,5-diphenylimidazole (DPI) in the proliferation of Brucella abortus infection. None of the compounds affected the uptake of Brucella into the macrophages. We determined the effect of neutralizing leukotriene B4 (LTB4) receptor and showed that the uptake of the bacteria was inhibited at 30 min post-infection. M4N treatment attenuated intracellular survival of Brucella at 2 h post-incubation but it was not observed in the succeeding time points. DPI treatment showed reduced survival of Brucella at 24 h post-incubation while blocking LTB4 receptor was observed to have a lower intracellular growth at 48 h post-incubation suggesting different action of the inhibitors in the course of the survival of Brucella within the cells. Reduced proliferation of the bacteria in the spleens of mice was observed in animals treated with ZIL or DPI. Increased serum cytokine level of TNF-α and MCP-1 was observed in mice treated with M4N or ZIL while a lower IFN-γ level in ZIL-treated mice and a higher IL-12 serum level in DPI-treated mice were observed at 7 d post-infection. At 14 d post-infection, ZIL-treated mice displayed reduced serum level of IL-12 and IL-10. Overall, inhibition of 5-LOX or TXA2 or a combination therapy promises a potential alternative therapy against B. abortus infection. Furthermore, strong ligands for LTB4 receptor could also be a good candidate for the control of Brucella infection.

Study on Motility and Acrosome Morphology of Fresh and Deep-frozen Korean Native Goat Spermatozoa (액상 및 동결보존된 한국재래산양 정자의 운동성 및 첨체형태에 관한 연구)

  • 황덕수;양문한;이규승;박창식
    • Korean Journal of Animal Reproduction
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    • v.13 no.1
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    • pp.18-25
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    • 1989
  • This study was carried out to investigate the general semen characteristics of the Korean native goat and the effect of temperature, incubation time, dilution rate, freezing rate and glycerol concentration on motility and NAR (normal apical ridge) acrosome of fresh and frozen Korean native goat spermatozoa. 1. Average semen volume per ejaculate, motility, concentration and pH of fresh Korean native goat spermatozoa were 0.19${\pm}$0.09 ml, 94.5${\pm}$0.47%, 26.17${\times}$108${\pm}$1.68/ml and 6.63${\pm}$0.18, respectively. 2. Motility and NAR acrosome of fresh spermatozoa during incubation were higher at 22$^{\circ}C$ than at 5$^{\circ}C$ or 37$^{\circ}C$(P<.01). 3. Motility and NAR acrosome of spermatozoa diluted 1:4 during incubation were higher at 22$^{\circ}C$ than at 5$^{\circ}C$ or 37$^{\circ}C$(P<.01). 4. Motility and NAR acrosome of spermatozoa during incubation were higher for samples diluted 1:1, 1:2, or 1:4 than for samples diluted 1:6(P<.01). 5. Motility and NAR acrosome of post-thaw spermatozoa were higher at freezing rate of 12$^{\circ}C$/min than at freezing rate of 1$^{\circ}C$/min or 24$^{\circ}C$/min when glycerol concentration was 9%(P<.01).

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A research for self-supporting management of business incubation center in region (지역 내 창업보육센터의 발전 방향에 대한 소고(小考))

  • Moon, Hyung-Joo;Choi, Kyo-Won;Hong, Ki-Hak;Lee, Gi-Sung;Son, Chang-Kyun
    • 한국벤처창업학회:학술대회논문집
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    • 2007.04a
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    • pp.313-328
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    • 2007
  • In this paper we investigated the organization of regional business incubation center itself and that of companies m center and scrutinized the problems and obstacles to take self-supporting administration for both of center and companies in it. We classified the companies operating inside the center into four types according to the formation of organizations analyzed them, and proposed some solutions to enhance their operating abilities to obtain elf-supporting administration.

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A research for self-supporting management of business incubation center in region (지역 내 창업보육센터의 발전 방향에 대한 소고(小考))

  • Moon, Hyung-Joo;Choi, Kyo-Won;Hong, Ki-Hak;Lee, Gi-Sung;Son, Chang-Kyun
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.2 no.1
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    • pp.1-14
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    • 2007
  • In this paper we investigated the organization of regional business incubation center itself and that of companies in center and scrutinized the problems and obstacles to take self-supporting administration for both of center and companies in it. We classified the companies operating inside the center into four types according to the formation of organizations analyzed them, and proposed some solutions to enhance their operating abilities to obtain elf- supporting administration.

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Effects of Incubation Temperatures on Hatching Period and Growth in Korea Reeves' Turtle(Mauremys Reevesii) (한국산 남생이의 부화와 성장에 부화 온도의 영향)

  • Jeong-Rae Rho
    • Korean Journal of Environment and Ecology
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    • v.37 no.3
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    • pp.192-197
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    • 2023
  • This study was conducted to determine whether the difference in egg hatching temperature of Korea Reeves' turtle (Mauremys reevesii) at the artificial nursery of Seoul National Grand Park affects the incubation period and growth. A total of 201 eggs were incubated at 26 ℃ (n=89), 28 ℃ (n=75) and 32 ℃ (n=37). The incubation period of eggs showed significant differences according to the hatching temperature. In this study, the higher the hatching temperature, the higher the hatching rate. The incubation period of the eggs hatched at 26 ℃, 28 ℃ and 32 ℃ was 66.1 (±4.0, n = 52) days, 65.3 (±3.3, n = 44) days and 58.8 (±7.7, n = 31) days, respectively. Eggs incubated at 32 ℃ (83.8%) had a higher hatching success than those at 26 ℃ (58.4%) and 28 ℃ (58.7%). The body mass of 14-day-old hatchlings incubated at 32 ℃ was greater than those incubated at 26 ℃ and 28 ℃. However, there was no significant difference in the mean body mass of 180 and 270-day-old turtles hatched at these different temperatures. This study showed that the hatching temperature significantly affected the incubation period and body mass in the early life of the Korea Reeves' turtle (M. reevesii).

Freeze-dried bovine amniotic membrane as a cell delivery scaffold in a porcine model of radiation-induced chronic wounds

  • Oh, Daemyung;Son, Daegu;Kim, Jinhee;Kwon, Sun-Young
    • Archives of Plastic Surgery
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    • v.48 no.4
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    • pp.448-456
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    • 2021
  • Background Locoregional stem cell delivery is very important for increasing the efficiency of cell therapy. Amnisite BA (Amnisite) is a freeze-dried amniotic membrane harvested from bovine placenta. The objective of this study was to investigate the retention of cells of the stromal vascular fraction (SVF) on Amnisite and to determine the effects of cell-loaded Amnisite in a porcine radiation-induced chronic wound model. Methods Initially, experiments were conducted to find the most suitable hydration and incubation conditions for the attachment of SVF cells extracted from pig fat to Amnisite. Before seeding, SVFs were labeled with PKH67. The SVF cell-loaded Amnisite (group S), Amnisite only (group A), and polyurethane foam (group C) were applied to treat radiation-induced chronic wounds in a porcine model. Biopsy was performed at 10, 14, and 21 days post-operation for histological analysis. Results Retaining the SVF on Amnisite required 30 minutes for hydration and 1 hour for incubation. A PKH67 fluorescence study showed that Amnisite successfully delivered the SVF to the wounds. In histological analysis, group S showed increased re-epithelialization and revascularization with decreased inflammation at 10 days post-operation. Conclusions SVFs had acceptable adherence on hydrated Amnisite, with successful cell delivery to a radiation-induced chronic wound model.