• Title/Summary/Keyword: Incubation Effect

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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.

In Vitro Development of Mouse Parthenogenetic Embryos: Effect of Temperature before Oocyte Activation

  • Roh Sangho;Won Cheolhee;Min Byung-Moo
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.117-120
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    • 2005
  • This study was conducted to establish the optimal temperature condition before oocyte activation in B6m F1 mouse. In experiment 1, two embryo culture media (CZB vs KSOM) were evaluated for the development of activated mouse oocytes. Parthenogenetic embryos cultured in KSOM showed better blastocyst development than ones cultured in CZB $(56.2\%\;vs\;81.0\%\;p<0.01)$. Two-hour of pre-incubation before activation significantly reduced the number of hatched blastocysts in KSOM $(22.0\%\;versus\;8.8\%\;p<0.05)$. In experiment 2, recovered oocytes were pre-incubated at different temperature conditions before activation. The experimental groups were divided by 5 as follows. Group A: pre-incubation for 120 min at $37^{\circ}C$, Group B: pre-incubation at $37^{\circ}C$ for 90 min then at $25^{\circ}C$ for 30 min, Group C: pre-incubation at $37^{\circ}C$ for 60 min then at $25^{\circ}C$for 60 min, Group D: pre-incubation at $37^{\circ}C$ for 30 min then at $25^{\circ}C$ for 90 min, and Group E: pre-incubation at $25^{\circ}C$ for 120 min before activation. Group A $(67.6\%)$ and B $(66.7\%)$ showed better development to the blastocyst stage than other groups $(Group\;C:\;50.0\%\;Group \;D:\;49.2\%\;Group\;E:\;33.3\%,\;p<0.05)$. The present study indicates that the temperature before activation affects the development of B6D2 F1 mouse parthenogenetic oocytes and exposure to room temperature should be limited to 30-min when the oocytes are left in HEPES-buffered medium for micromanipulation.

EFFECT OF SUPPLEMENTARY UREA, GLUCOSE AND MINERALS ON THE IN VITRO DEGRADATION OF LOW QUALITY FEEDS

  • Oosting, S.J.;Verdonk, J.M.H.J.;Spinhoven, G.G.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.2 no.4
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    • pp.583-590
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    • 1989
  • Increasing levels of ammonia-N in the rumen fluid used for in vitro incubation were achieved by supplementation of the ration of the donor cows with urea and by addition of urea either with or without glucose to the rumen fluid after collection. The ration of the donor animals consisted of wheat straw (80%) and maize silage (20%). During the second half of the experiment the basal ration was supplemented with a mineral mixture. Wheat straw, Guinea grass and two rice straw varieties were incubated with the various kinds of rumen fluid. Parameters studied were: solubility, apparent organic matter disappearance after 48 hours of incubation ($OMD_{48}$), rate of organic matter degradation from 0 to 24 hours of incubation ($k_1$) and from 24 to 95 hours ($k_2$). The concentration of ammonia-N in the rumen fluid at which 95% of the maximal $OMD_{48}$ and k1 were reached (88.2 and 100.0 mg/l) were independent of the feed. With regard to the $k_2$ the required ammonia-N concentration to reach 95% of the maximal $k_2$ differed per feed. Mineral supplementation increased the OMD48 and $k_1$, but not the solubility and $k_2$. Glucose addition in combination with urea had no beneficial effect compared to urea supplementation alone.

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.

Effect of Soil Water Contents on Urea Hydrolysis and Nitrification in a Newly Reclaimed Tidal Soils

  • Park, Mi-Suk;Kim, Hye-Jin;Chung, Doug-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.48-52
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    • 2011
  • The effect of soil water content on the transformation potential of N compounds derived from hydrolysis of urea applied in a reclaimed tidal soils which was saline-sodic was observed to evaluate nitrification rates of urea. Soil samples were collected from Moonpo series at the newly reclaimed area in Saemanguem. For the transformation potential of N compounds from urea (46% N), newly reclaimed tidal soils (RS) were amended with urea at the rates of 0, 10, and 20 kg $10a^{-1}$. With leachate obtained from the incubated RS in a leaching tube at $25^{\circ}C$, urea hydrolysis and nitrification were measured for a total of 30days. The cumulative amounts of $NO_3{^-}$-N in each of the four soils treated with urea was linear with time of incubation. Results showed that increase in pH occurred with increasing application rate of urea and volumetric water content due to hydrolysis of urea. The total N in the RS was decreased with incubation time, indicating that rates of urea hydrolysis was influenced by soil moisture conditions. Also, the cumulative amount of nitrate in RS gradually increased with increase in time of incubation.

AUTOLYSIS IN THE ANAEROBIC FUNGUS Piromyces communis OTS1 : PRESENCE OF CHITINASE AND β-1, 3-GLUCANASE ACTIVITIES

  • Sakurada, M.;Morgavi, D.P.;Ohishi, T.;Onodera, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.3
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    • pp.303-307
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    • 1996
  • The degree of autolysis and presence of cell-wall degrading enzymes in an anaerobic ruminal fungus, Piromyces communis OTSI, grown in liquid medium, was monitored to evaluate the effect of self-digestion on fungal biomass. After a 30 days incubation period fungal dry weight decreased by 45% and the cell wall component chitin decreased by 22%. Chitinase activity detected in the supernatant was mainly of the endotype and peaked at day 6 of the incubation. ${\beta}-1$, 3-glucanase was detected from day 4 and increased throughout the incubation period. Autolysis was a slow process, and under natural conditions it is unlikely that it plays a significant role in the degradation of the spent fungal vegetative stage in the rumen.

Fungal bioconversion of Korean food wastes for the production of animal feed additive enzymes

  • Jeong, Yun-Seung;Jeong, Sang-Won;Jo, A-Ra;Gwon, Sun-U;Han, Seung-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.529-532
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    • 2001
  • Korean food waste, one of the abundantly available but environmentally problematic organic wastes in Korea, was utilized as solid-substrate by fungal strain Aspergillus niger ATcC 6275 for the production of enzymemixture containing amylase, cellulase and xylanase. The enzyme mixture can be used as high value-added animal feed. Solid-state fermentation method yielded a 84-fold enhancement in xylanase activity compared with submerged fermentation method. The effect of incubation period, incubation temperature, pH of medium, moisture content, inoculum size and enrichment of the medium with nitrogen and carbon sources were observed for optimal production of these enzymes The optimal amylase activity of 33.10 U/g, cellulase activity of 24.41 U/g, xylanase activity of 328.84 U/g were obtained at 8 days incubation with 50%(w/w) soy bean flake, with incubation temperature of $25^{\circ}C$, pH of 6.38, optimal moisture content of 55% and with inoculum size of $3.8{\times}10^6$spore/g. Enzyme activities were enhanced when ImM $CaSO_4$, 2% Malt extract and 2% galactose were added as mineral, nitrogen and carbon enrichment respectively.

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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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Effect of Growth Hormone on Vitellogenin Production by Estradiol-17$\beta$ in the Culture of Hepatocytes in the Rainbow Trout Oncorhynchus mykiss

  • Yeo In-Kyu;Mugiya Yasuo
    • Fisheries and Aquatic Sciences
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    • v.1 no.1
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    • pp.19-23
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    • 1998
  • Effects of pituitary and thyroid hormones on estradiol-induced vitellogenin (VTG) induction were electrophoretically examined in primary hepatocyte cultures of rainbow trout. Hepatocytes were precultured for 2 days and then estradiol-17 $\beta$ $(E_2,\;2 \times 10^{-6}M)$>, triiodothyronine $(T_3,\;10^{-8}-10^{-6}M)$, bovine growth hormone (bGH, 10-100 ng/ml), ovine prolactin (oPRL, 100-500 ng/ml), and pituitary extract (PE) of rainbow trout (0.75PE/dish) were added to the incubation medium. The hepatocytes were cultured for 7 more days. The addition of oPRL to the incubation medium was not effective in increasing VTG production at any concentrations. The addition of PE to the incubation medium with $E_2$ was not effective in increasing VTG production. The addition of bGH to the incubation medium with $E_2$ was not effective in increasing the rate of VTG production at concentrations of 10-50 ng/ml. However, a higher concentration of bGH, 100 ng/ml, increased VTG production. The various concentrations of $T_3$ were ineffective in stimulating VTG production. These results suggest that GH could be one of stimulus factors for VTG production in rainbow trout.

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Some Aspects of Laying, Incubation and Hatching in the Great Reed-Warbler

  • Yoo, Jeong-Chil;Park, Yu-Seong
    • 한국생태학회:학술대회논문집
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    • 2002.08a
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    • pp.107-111
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    • 2002
  • During the breeding season of 1998, breeding ecology of the Great Reed-Warbler (Acrocephalus arundinaceus orientalis) was studied at Yangsoo-ri and Yongdam-ri of the Yangpyung-gun, Kyunggi province, Korea. Egg-weight (CV: 6.25) was more variable than either length or breadth, and breadth was the least variable of the measures. Significant variations In overall egg-weight occurred between clutches, and that more of the total variation in egg-weight and shape are due to inter-clutch variation as to intra-clutch variation when the data were pooled. The last egg tends to be larger than the remaining eggs in the clutch of the Great Reed-Warbler, suggest- ing the Great Reed-Warbler may adopt the brood-survival strategy. When method 3 was used, the most common incubation period is 12 days. In the Great Reed-Warbler, the length of the incubation period was related to clutch-size when method 1 (r=0.485, p<0.05) and method 2 (r=0.621, p<0.01) were employed, but not related to egg weight. The average number of days of hatching asynchrony was 2.5, raging 0.5∼2.5. Asynchronous hatching was related to the clutch size (r=0.66, p<0.01). Hatching sequence was closely related to the laying sequence (r=0.93, p<0.001), suggesting Great Reed-Warblers incubate their eggs before clutch completion. The effect of egg weight on hatching asynchrony was found in Great Reed-Warblers (t-test, p<0.01).

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