• Title/Summary/Keyword: Co-incubation

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Effect of Incubation Time after Cooling on the Meiotic Spindle and Chromosomes of Mouse Oocytes (냉각 후 배양시간이 생쥐 난자의 방추체와 염색체에 미치는 영향)

  • Yu I.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.283-289
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    • 2004
  • This study was conducted to determine the effects of incubation time after cooling on mouse meiotic spindle and chromosome alignment and the optimal incubation time for their restoration. Oocytes at the metaphase II were obtained from superovulated mice. Control oocytes were held at 37$^{\circ}C$ during the experiment. Oocytes were rapidly cooled to $0^{\circ}C$, held for 30 minutes, warmed and incubated at 37$^{\circ}C$ for 5, 15, 30, 60 and 120 minutes, respectively. The morphological features of spindle and chromosomes in oocytes were evaluated by immunofluorescent staining. Meiotic spindle of control oocytes exhibited a normal-looking bipolar configuration(barrel-shaped) and highly fluorescent microtubles. The chromosomes were clustered in a discrete bundles at metaphase plate. Disassembly of meiotic spindle and chromosome dispersion were occurred immediately after chilling of oocyte. Fluorescence intensity index(FIS), normal chromosomes aligned and normal spindle configuration were compared according to incubation time at 37$^{\circ}C$. Restoration of a barrel-shaped spindle and normal chromosome alignment was occurring after 5 minutes incubation at 37$^{\circ}C$, improved as a incubation time increased, and decreased gradually after 120 minutes incubation(P<0.05). The optimal incubation time for restoration of meiotic spindle and chromosomes in cooled oocytes was 60 minutes.

Effects of Defaunation on Fermentation Characteristics, Degradation of Ryegrass Hay and Methane Production by Rumen Microbes In Vitro When Incubated with Plant Oils

  • Qin, Wei-Ze;Li, Cheng-Yun;Choi, Seong-Ho;Jugder, Shinekhuu;Kim, Hyun-Ju;Lee, Sang-Suk;Song, Man-Kang
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.34 no.3
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    • pp.193-201
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    • 2014
  • This study was conducted to examine the effects of defaunation (removal of live protozoa) on fermentation characteristics, degradation of ryegrass hay and $CH_4$ (methane) production by rumen microbes when incubated with plant oils (SO, sunflower oil and LO, linseed oil) in vitro. Sodium lauryl sulfate (0.000375 g/ml) as a defaunation reagent was added into the culture solution and incubated anaerobically up to 24 h at $39^{\circ}C$. pH from defaunation was increased for all treatments from 6 h incubation times (p<0.01-0.001) compared with those from fauantion. Concentration of ammonia-N from defaunation is higher than that from faunation at 3 h (p<0.001), 12 h (p<0.05) and 24 h (p<0.001) incubation times. Defaunation decreased (p<0.01-0.001) total volatile fatty acid concentration at all incubation times. Molar proportions of $C_2$ (acetate, p<0.05-0.001) and butyrate (p<0.01-0.001) were also decreased by defaunation at all incubation times. Molar proportion of $C_3$ (propionate), however, was increased by defaunation at all incubation times (p<0.001). Thus the rate of $C_2$ to $C_3$ was decreased by defaunation at all incubation times (p<0.001). Defaunation decreased ED (effective degradability) of dry matter (p<0.001) and ED of neutral detergent fiber (p<0.001) of ryegrass hay. Defaunation decreased total gas, $CH_4$ production, $CH_4$ % in total gas and $CH_4/CO_2$ at all incubation times (p<0.001). Oil supplementation decreased total gas (p<0.05-0.001), $CH_4$ production (p<0.001) and $CH_4$ % in total gas (p<0.001) compared with control at all incubation times. The result of this study showed that defaunation combined with oil supplementation may cause an alteration of microbial communities and further medicate the fermentation pattern, resulting in both reduction of degradation of ryegrass hay and $CH_4$ production. No difference, however, was observed in all the examinations between SO and LO.

Rumen fermentation, methane production, and microbial composition following in vitro evaluation of red ginseng byproduct as a protein source

  • Hamid, Muhammad Mahboob Ali;Moon, Joonbeom;Yoo, Daekyum;Kim, Hanbeen;Lee, Yoo Kyung;Song, Jaeyong;Seo, Jakyeom
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.801-811
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    • 2020
  • The main objective of this in vitro study was to evaluate red ginseng byproduct (RGP) as a protein resource and its effects on rumen fermentation characteristics, microflora, CO2, and CH4 production in ruminants. Four treatments for in vitro fermentation using buffered rumen fluid over a 48 h incubation period were used: 1, RGP; 2, corn gluten feed (CGF); 3, wheat gluten (WG); and 4, corn germ meal. In vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), in vitro crude protein digestibility (IVCPD), volatile fatty acids, pH, and ammonia nitrogen (NH3-N) were estimated after 48 h incubation. Gas production was investigated after 3, 6, 12, 24, 36 and 48 h. The CO2 and CH4 were evaluated after 12, 24, 36, and 48 h. A significant difference in total gas production and CO2 emissions was observed (p < 0.01) at all incubation times. CH4 production in RGP were higher (p < 0.05) than that in other treatments but a higher CH4 portion in the total gas production was observed in WG (p < 0.05) at 48 h incubation. The IVDMD, IVNDFD, and IVCPD of RGP was lower than those of other conventional ingredients (p < 0.01). The RGP had the lowest NH3-N value among the treatments (p < 0.01). The RGP also had the lowest total VFA concentration (p < 0.01), but presented the highest acetate proportion and acetate to propionate ratio among the treatments (both, p < 0.01). The abundance of Prevotella ruminicola was higher in RGP than in WG (p < 0.01), whereas RGP has lower methanogenic archaea (p < 0.01). In conclusion, based on the nutritive value, IVDMD, low NH3-N, and decreased methanogenic archaea, RGP inclusion as a protein source in ruminant diets can be an option in replacing conventional feed sources.

Influence of Transition-Metal Cofactors on the Reductive Dechlorination of Polychlorinated Biphenyls (PCBs)

  • Kwon, O-Seob;Kim, Young-Jin;Cho, Kyung-Je;Lee, Jin-Ae;Kim, Young-Eui;Hwang, In-Young;Kwon, Jae-Hyun
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.189-195
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    • 2003
  • To enhance the reductive dechlorination of polychlorinated biphenyls (PCBs) under anaerobic conditions, we examined the adjunctive effects of cobalt (Co) and nickel (Ni), which are the central metals of transition-metal cofactors of coenzyme F$\_$430/ and vitamin B$\_$12/, respectively, on the dechlorination of Aroclor 1248. After 32 weeks of incubation, the average numbers of chlorines per biphenyl in culture vials supplemented with 0.2, 0.5, and 1.0 mM of Co reduced from 3.88 to 3.39, 2.92, and 3.28, respectively. However, the numbers of chlorine after supplementing with Ni decreased from 3.88 to 3.43, regardless of the Ni concentrations. The observed congener distribution patterns of all vials with different conditions were similar to the pattern produced by the dechlorination process of H' after 21 weeks of incubation, and these patterns were unchanged up to week 32, except for vials supplemented with 0.5 and 1.0 mM of Co. In vials containing 0.5 mM of Co, meta-rich congeners, such as 25/ 25-,24/25-, and 25/23-chlorobiphenyls (CBPs), which were found as accumulated products of dechlorination in other conditions, were further dechlorinated, and 25/2-, 24/2-, and 2/2-CBPs were concomitantly increased after 32 weeks of incubation. In this case, the congener distribution was similar to the dechlorination pattern of process M. From these results, we suggested that the enrichment of cultures with Co might stimulate the growth of specific populations of meta-dechlorinators, and that populations might promote a change in the dechlorination process from H' to M, which is known to be less effective on the dechlorination of the more highly chlorinated congeners of PCBs.

Effects of various Pretreatments on the Nucleation of CVD Tungsten (전처리가 CVD 텅스텐의 핵 생성에 미치는 영향)

  • Kim, Eui-Song;Lee, Chong-Mu;Lee, Jong-Gil
    • Korean Journal of Materials Research
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    • v.2 no.6
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    • pp.443-451
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    • 1992
  • Effects of various pretreatments on the nucleation of CVD-W deposited on the reactively sputter-deposited TiN was investigated. Incubation period of nucleation and deposition rate decreased by the pretreatment of Ar rf-sputter etching for the depth below 300k, but they increased for the etchig depth over 200A. The preteatment of Ar ion implantation decreased the incubation period of nucleation, but increased deposition rate. Also Si$H_4$flushing pretreatment decreased the incubation period of nucleation slightly due to the absorption of Si by TiN surface.

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Antitumor Activity of the Novel Human Cytokine AIMP1 in an in vivo Tumor Model

  • Lee, Yeon-Sook;Han, Jung Min;Kang, Taehee;Park, Young In;Kim, Hwan Mook;Kim, Sunghoon
    • Molecules and Cells
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    • v.21 no.2
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    • pp.213-217
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    • 2006
  • Although AIMP1 (previously known as p43) is one of three auxiliary proteins bound to a macromolecular aminoacyl tRNA complex, it is also secreted as a cytokine controlling both angiogenesis and immune responses. Here we show that systemically administered purified recombinant human AIMP1 had anti-tumor activity in mouse xenograft models. In Meth A-bearing Balb/c mice, tumor volume increased about 28 fold in the vehicle treatment group, while an increase of about 16.7 fold was observed in the AIMP1-treated group. We also evaluated the anti-tumor activity of AIMP1 in combination with a sub-clinical dose of the cytotoxic anti-tumor drug, paclitaxel. The growth of NUGC-3 human stomach cancer cells was suppressed by 84% and 94% by the combinations of 5 mg/kg paclitaxel + 25 mg/kg AIMP1 (p = 0.03), and 5 mg/kg paclitaxel + 50 mg/kg AIMP1 (p = 0.02), respectively, while 5 mg/kg paclitaxel alone suppressed growth by only 54% (p = 0.02). A similar cooperative effect of AIMP1 and paclitaxel was observed in a lung cancer xenograft model. These results suggest that AIMP1 may be useful as a novel anti-tumor agent.

Curcumin-Induced Autophagy Augments Its Antitumor Effect against A172 Human Glioblastoma Cells

  • Lee, Jong-Eun;Yoon, Sung Sik;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.27 no.5
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    • pp.484-491
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    • 2019
  • Glioblastoma is the most aggressive common brain tumor in adults. Curcumin, from Curcuma longa, is an effective antitumor agent. Although the same proteins control both autophagy and cell death, the molecular connections between them are complicated and autophagy may promote or inhibit cell death. We investigated whether curcumin affects autophagy, which regulates curcumin-mediated tumor cell death in A172 human glioblastoma cells. When A172 cells were incubated with $10{\mu}M$ curcumin, autophagy increased in a time-dependent manner. Curcumin-induced cell death was reduced by co-incubation with the autophagy inhibitors 3-methyladenine (3-MA), hydroxychloroquine (HCQ), and LY294002. Curcumin-induced cell death was also inhibited by co-incubation with rapamycin, an autophagy inducer. When cells were incubated under serum-deprived medium, LC3-II amount was increased but the basal level of cell viability was reduced, leading to the inhibition of curcumin-induced cell death. Cell death was decreased by inhibiting curcumin-induced autophagy using small interference RNA (siRNA) of Atg5 or Beclin1. Therefore, curcumin-mediated tumor cell death is promoted by curcumin-induced autophagy, but not by an increase in the basal level of autophagy in rapamycin-treated or serum-deprived conditions. This suggests that the antitumor effects of curcumin are influenced differently by curcumin-induced autophagy and the prerequisite basal level of autophagy in cancer cells.

Variation Patterns in Concentration of Inorganic Nitrogen from Liquid Grass Fertilizer during Aerobic Incubation (항온 호기 배양 조건에서 잔디 예초물 액비로부터 무기화된 질소의 농도 변화)

  • Lee, Tae-Kyu;Park, Ji-Suk;Lee, Min-Jin;Kim, Jong-Sung;Ro, Hee-Myong;Kim, Sang-Jun;Jeon, Seung-Woo;Seo, Sang-Gug;Kim, Kil-Yong;Lee, Geon-Hyoung;Jeong, Byung-Gon
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1120-1125
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    • 2012
  • To assess fertilizer value of an quasi-aerobically fermented liquid clipped-grass fertilizer, aerobic incubation experiment using two texturally contrasting loam (L) and sandy loam (SL) soils was conducted for 60 days to investigate temporal variations in N mineralization pattern of the liquid fertilizer applied. To do so, the quasi-aerobically fermented liquid clipped-grass fertilizer was prepared, applied to each soil at a rate of 200 kg-N $ha^{-1}$ and aerobically $25^{\circ}C$ in the dark. During incubation, soil water content was adjusted to field moisture capacity (-33 kPa of soil matric potential) by adding distilled water as necessary to maintain their initial weights. At desired time of incubation (0, 1, 5, 10, 20, 40, and 60 days after incubation), soil was sampled and analyzed for inorganic nitrogen ($NH_4{^+}$-N and $NO_3{^-}$-N) concentrations, pH, EC, total carbon contents and total nitrogen contents. Concentrations of $NH_4{^+}$-N began to decrease right after incubation for L soils, and 10 days after incubation for SL soils, while those of $NO_3{^-}$-N began to increase onset of $NH_4{^+}$-N disappearance. The results of this study showed that quasi-aerobically fermented liquid clipped-grass fertilizer could serve as an alternative to chemical N fertilizer.

Fermented Soymilk Alleviates Lipid Accumulation by Inhibition of SREBP-1 and Activation of NRF-2 in the Hepatocellular Steatosis Model

  • Ahn, Sang Bong;Wu, Wen Hao;Lee, Jong Hun;Jun, Dae Won;Kim, Jihyun;Kim, Riji;Lee, Tae-bok;Jun, Jin Hyun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.236-245
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    • 2018
  • Ingredients of soy and fermented soy products have been widely utilized as food supplements for health-enhancing properties. The aim of this study was to evaluate the effects of fermented soymilk (FSM) and soymilk (SM) on free fatty acid-induced lipogenesis in the hepatocellular steatosis model. HepG2 cells were incubated with palmitic acid (PA) for 24 h to induce lipogenesis and accumulation of intracellular lipid contents. The PA-treated cells were co-incubated with FSM, SM, genistein, and estrogen, respectively. Lipid accumulation in the PA-treated HpG2 cells was significantly decreased by co-incubation with FSM. Treatment of HepG2 cells with PA combined with genistein or estrogen significantly increased the expression of SREBP-1. However, FSM co-incubation significantly attenuated SREBP-1 expression in the PA-treated HepG2 cells; in addition, expression of NRF-2 and phosphorylation of ERK were significantly increased in the PA and FSM co-incubated cells. PA-induced ROS production was significantly reduced by FSM and SM. Our results suggested that the bioactive components of FSM could protect hepatocytes against the lipid accumulation and ROS production induced by free fatty acids. These effects may be mediated by the inhibition of SREBP-1 and the activation of NRF-2 via the ERK pathway in HepG2 cells.