• Title/Summary/Keyword: culture-dependent

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Oocyte-sperm Binding Assay (OSBA) Technique for Rapid Q/C of IVF Culture Condition (체외수정용 배양조건의 신속한 Q/C를 위한 정자-난자 결합분석법(OSBA) 개발)

  • 정구민;신영수
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.163-169
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    • 2001
  • OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P < 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

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Trend and Further Research of Rice Quality Evaluation (쌀의 품질평가 현황과 금후 연구방향)

  • Son, Jong-Rok;Kim, Jae-Hyun;Lee, Jung-Il;Youn, Young-Hwan;Kim, Jae-Kyu;Hwang, Hung-Goo;Moon, Hun-Pal
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47
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    • pp.33-54
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    • 2002
  • Rice quality is much dependent on the pre-and post harvest management. There are many parameters which influence rice or cooked rice qualitys such as cultivars, climate, soil, harvest time, drying, milling, storage, safety, nutritive value, taste, marketing, eating, cooking conditions, and each nations' food culture. Thus, vice evaluation might not be carried out by only some parameters. Physicochemical evaluation of rice deals with amy-lose content, gelatinizing property, and its relation with taste. The amylose content of good vice in Korea is defined at 17 to 20%. Other parameters considered are as follows; ratio of protein body-1 per total protein amount in relation to taste, and oleic/linoleic acid ratio in relation to storage safety. The rice higher Mg/K ratio is considered as high quality. The optimum value is over 1.5 to 1.6. It was reported that the contents of oligosaccharide, glutamic acid or its derivatives and its proportionalities have high corelation with the taste of rice. Major aromatic compounds in rice have been known as hexanal, acetone, pentanal, butanal, octanal, and heptanal. Recently, it was found that muco-polysaccharides are solubilized during cooking. Cooked rice surface is coated by the muco-polysaccharide. The muco-polysaccharide aye contributing to the consistency and collecting free amino acids and vitamins. Thus, these parameters might be regarded as important items for quality and taste evaluation of rice. Ingredients of rice related with the taste are not confined to the total rice grain. In the internal kernel, starch is main component but nitrogen and mineral compounds are localized at the external kernel. The ingredients related with taste are contained in 91 to 86% part of the outside kernel. For safety that is considered an important evaluation item of rice quality, each residual tolerance limit for agricultural chemicals must be adopted in our country. During drying, rice quality can decline by the reasons of high drying temperature, overdrying, and rapid drying. These result in cracked grain or decolored kernel. Intrinsic enzymes react partially during the rice storage. Because of these enzymes, starch, lipid, or protein can be slowly degraded, resulting in the decline of appearance quality, occurrence of aging aroma, and increased hardness of cooked rice. Milling conditions concerned with quality are paddy quality, milling method, and milling machines. To produce high quality rice, head rice must contain over three fourths of the normal rice kernels, and broken, damaged, colored, and immature kernels must be eliminated. In addition to milling equipment, color sorter and length grader must be installed for the production of such rice. Head rice was examined using the 45 brand rices circulating in Korea, Japan, America, Australia, and China. It was found that the head rice rate of brand rice in our country was approximately 57.4% and 80-86% in foreign countries. In order to develop a rice quality evaluation system, evaluation of technics must be further developed : more detailed measure of qualities, search for taste-related components, creation and grade classification of quality evaluation factors at each management stage of treatment after harvest, evaluation of rice as food material as well as for rice cooking, and method development for simple evaluation and establishment of equation for palatability. On policy concerns, the following must be conducted : development of price discrimination in conformity to rice cultivar and grade under the basis of quality evaluation method, fixation of head rice branding, and introduction of low temperature circulation.

Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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Characterization of Bacteriocin Produced from Isolated Strain of Bacillus sp. (Bacillus 속 분리주가 생산하는 박테리오신의 특성 조사)

  • Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.202-210
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    • 2017
  • As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

Synthetic Chenodeoxycholic Acid Derivative HS-1200-Induced Apoptosis of Human Oral Squamous Carcinoma Cells (합성 Chenodeoxycholic Acid 유도체 HS-1200이 유도한 사람구강 편평상피암종세포 세포자멸사 연구)

  • Kim, In-Ryoung;Sohn, Hyeon-Jin;Kim, Gyoo-Cheon;Kwak, Hyun-Ho;Park, Bong-Soo;Choi, Won-Chul;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.32 no.3
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    • pp.251-261
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    • 2007
  • Bile acids and synthetic its derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. Previous studies have been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis inducing activity on various cancer cells in vitro. It wasn't discovered those materials have apoptosis induced effects on YD9 human oral squamous carcinoma cells. The present study was done to examine the synthetic bile acid derivatives(HS-1199, HS-1200) induced apoptosis on YD9 cells and such these apoptosis events. We administered them in culture to YD9 cells. Tested YD9 cells showed several lines of apoptotic manifestation such as activation of caspase-3, degradation of DFF, production of poly (ADP-ribose) polymerase(PARP) cleavage(HS-1200 only), DNA degradation(HS-1200 only), nuclear condensation, inhibition of proteasome activity, reduction of mitochondrial membrane potential(HS-1200 only) and the release of cytochrome c and AIF to cytosol. Between two synthetic CDCA derivatives, HS-1200 showed stronger apoptosis-inducing effect than HS-1199. Therefore HS-1200 was demonstrated to have the most efficient antitumor effect. Taken collectively, we demonstrated that a synthetic CDCA derivative HS-1200 induced caspases-dependent apoptosis via mitochondrial pathway in human oral sqauamous carcinoma cells in vitro. Our data therefore provide the possibility that HS-1200 could be considered as a novel therapeutic strategy for human orall squamous carcinoma from its poweful apoptosis-inducing activity.

Protective Effects of Enzymatic Oyster Hydrolysate on Acetaminophen-induced HepG-2 Cell Damage (아세트아미노펜 유도 HepG-2 세포주 손상에 대한 굴 효소 가수분해물의 보호 효과)

  • Park, Si-Hyang;Moon, Sung-Sil;Xie, Cheng-Liang;Choung, Se-Young;Choi, Yeung-Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1166-1173
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    • 2014
  • This study investigated the detoxification effects of enzymatic hydrolysate from oyster on acetaminophen-induced toxicity using HepG-2 cells. Oyster hydrolysate was made with 1% Protamex and 1% Neutrase after treatment with transglutaminase (TGPN) or without (PN). Two types of oyster hydrolysate were added to human-derived HepG-2 hepatocytes damaged by acetaminophen, after which the survival rate of HepG-2 cell was measured. In addition, glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) activities in the culture media were evaluated. The survival rates of HepG-2 cells were $136.2{\pm}1.4%$ at $100{\mu}g/mL$ of TGPN and $179.6{\pm}3.8%$ at $200{\mu}g/mL$ of TGPN. These cell survival rates were higher compared to that of the negative control group ($60.7{\pm}3.2%$) treated only with acetaminophen. GOT activity was $38.3{\pm}0.2$ Karmen/mL in the negative control group, whereas it was $19.9{\pm}0.5$ for TGPN ($200{\mu}g/mL$) and $22.0{\pm}2.4$ Karmen/mL for PN ($200{\mu}g/mL$). GOT and GTP activities were shown to be dependent on TGPN concentration, and significant reduction in activities could be conformed. The detoxification efficacy of TGPN was higher compared to that of PN. These results suggest that oyster hydrolysate has potential as a healthy food or pro-drug for liver protection.

Inhibitory effects of heavy metals on CYP1A expression in eel hepatocyte cultures (뱀장어 배양 간세포에서의 Cytochrome P4501A (CYP1A) 유전자 발현에 대한 중금속들의 억제효과)

  • Kwon, Hyuk-Chu;Maeng, Joon-Ho;Choi, Seong-Hee
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.245-254
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    • 2010
  • Effects of heavy metal ions on the gene expression of cytochrome P4501A (CYP1A) were examined in cultured eel hepatocytes. When the expression of CYP1A mRNA was measured by RT-PCR after incubation of eel hepatocytes with benzo[$\alpha$]pyrene (B[$\alpha$]P) at concentrations of 10-8~10-5 M, the CYP1A expression increased with B[$\alpha$]P treatment in a dose dependent manner, showing significant increase at concentrations more than 10-7 M. When the eel hepatocyte was treated with cadmium (10-6 and 10-5 M), the expression of CYP1A was inhibited and especially at higher concentration (10-5 M). The inhibition of CYP1A expression by cadmium was also observed in cells treated with B[$\alpha$]P. In another study, effects of heavy metal ions on the expression of CYP1A were examined in cultured hepatocytes isolated from eel which was treated previously with B[$\alpha$]P in vivo. Hepatocytes isolated from the liver of eel taken at 48 hours after injection of B[$\alpha$]P (10 mg/kg) were cultured for 2 days with cadmium, copper, lead or zinc (10-6 and 10-5 M). The expression of CYP1A was found to be suppressed by the metal ions compared with the control in which CYP1A was induced with previous treatment of B[$\alpha$]P in vivo. The present results may provide an important basic information for studying the effects of heavy metal ions on CYP1A expression in other species of fish and studying toxicological mechanisms of heavy metal ions in aquatic livings.

Reduction of dissolved hydrogen sulfide and mortality of white leg shrimp, Litopenaeus vannamei by Bacillus spp. microorganisms (Bacillus속 미생물의 용존황화수소 저감효과와 흰다리새우(Litopenaeus vannamei)에의 영향)

  • Choi, Jun-Ho;Lee, Ji-Hoon;Park, Jung-Jin;Lee, Min-Sun;Bae, Jun-Sung;Shin, Dong-Hun;Park, Kwan Ha
    • Journal of fish pathology
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    • v.31 no.1
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    • pp.41-48
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    • 2018
  • The utility of Bacillus spp. organisms for reduction of dissolved hydrogen sulfide ($H_2S$) in white leg shrimp (Litopenaeus vannamei) culture was tested with different combinations of Bacillus spp. microorganisms: combination A (B. subtilis + B. licheniformis); combination B (B. licheniformis + B. amyloliquefaciens); combination C (B. subtilis + B. licheniformis + B. amyloliquefaciens). Of these 3 combinations, C was effective in few hours after addition whereas B needed longer time to be effective. The $H_2S-reducing$ effect of combination C was dependent on the amount of microorganisms added to $H_2S-containing$ test solution. Exposure of white leg shrimp to $H_2S$ at 8 mg/L for 7 days led to survival of 80% and 1 mg/L for 14 days it was 82.5%. The survival rate was 97.5% when combination C was simultaneously added to shrimp tanks during $H_2S$ exposure at 1 mg/L for 14 days. It was demonstrated that combination C microorganisms (B. subtilis + B. licheniformis + B. amyloliquefaciens) can reduce dissolved $H_2S$ concentrations, and this effect can be utilized to protect white leg shrimp from $H_2S$ toxicity.

Preparation and Quality Characteristics of the Fermentation product of Ginseng by Lactic Acid Bacteria (FGL) (유산균을 이용한 발효인삼 제조 및 품질 특성)

  • Park, Soo-Jin;Kim, Dong-Hyun;Paek, Nam-Soo;Kim, Sung-Soo
    • Journal of Ginseng Research
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    • v.30 no.2
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    • pp.88-94
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    • 2006
  • Ginseng as a raw material for production of probiotic ginseng product by lactic acid bacteria (LAB) was evaluated in this study. Either white ginseng (WG) or red ginseng (RG) (1% or 5%, w/v) were directly inoculated with a 24 hold seed culture of twenty seven substrains of four different LAB ($1.0{\times}10^6CFU/ml$); Lactobacillus spp., Streptococcus/Enterococcus spp., Leuconostoc/Lactococcus spp. and Bifidobacterium spp., and incubated at $37^{\circ}C$ for 24 or 48 h. Among 27 kinds of LAB, seven substrains of Lactobacillus (MG208, MG311, MG315, MG501, MG501C, MG505, MG590) and one Bifidobacterium (MG723) were selected based on their dose dependent stimulation of the growth of LAB in the presence of ginseng and changes in pH, acidity and viable cell counts during fermentation were examined. Lactobacillus MG208 specifically was found to show the best growth on 5% RG and reached nearly $14.0{\times}10^8CFU/ml$ after 48 h of fermentation and produced the titratable acidity as $0.84{\pm}0.02%$, whereas the pH was significantly lowered from $6.80{\pm}0.01\;to\;3.42{\pm}0.02$. These results indicated that ginseng can be an appropriate material to prepare the fermentation product by several strains of LAB. Therefore we should further check whether probiotic ginseng product may have synergistic health benefits of both probiotics and ginseng to serve for vegetarians and lactose-allergic consumers.

Moderating Effect of Technology Development Activities Among Entrepreneurial Orientation, the Capability of Technology Innovation and Commercialization Performance: Focused on ICT Technology New Ventures (기술개발활동의 기업가적 지향성, 기술혁신역량과 기술사업화 성과와의 관계에서 조절적 효과 분석: ICT 창업기업을 중심으로)

  • Kim, Chang-Bong;Bae, Keun-Suk
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.16 no.5
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    • pp.31-47
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    • 2021
  • The purpose of this study is to demonstrate the moderating effect of technology development activities in the relationship between independent variables such as entrepreneurial orientation and technology innovation capabilities and dependent variables. As a result of analyzing the causal relationship between research variables, it was found that the higher the innovation and initiative among the sub-factors of entrepreneurial orientation, the more positive the technical commercialization performance and product completion. Among the sub-factors of entrepreneurial orientation, risk-taking was found to have a significant effect only on product completion. It was found that the higher the technology commercialization capability and technology convergence capability, the higher the technology commercialization performance, the technology commercialization performance. As a result of analyzing the moderating effect of technology development activities, it was found that technology development management ability, a sub-factor of technology development activities, controls the influence relationship between innovation and risk sensitivity and technology performance. In addition, it was found that the involvement in technology development planning controls the influence relationship between technology convergence capability and technology performance among sub-factors of technology innovation capability. Based on the above analysis results, this study made three suggestions as follows. First, the achievements of technology commercialization to achieve the superiority of corporate competition depend on progressive innovation and risk-taking based on entrepreneurial orientation. It is necessary to find a way to build entrepreneurial orientation from within the organization. Second, due to the nature of the ICT industry, which has a fast pace of technological development and changes in market acceptance, technology commercialization performance will be positive when the capabilities, technology, knowledge, and resources that can quickly lead to product production can be organically linked. Finally, corporate CEOs need to further promote innovation and risk-taking through phased and continuous research activities for technology development. In addition, it is necessary to establish a corporate culture that tolerates various strategies and failures so that understanding of technology convergence can lead to technological performance.