• Title/Summary/Keyword: beta type

Search Result 2,080, Processing Time 0.028 seconds

Secretory Proteins from Goat Oocytes Matured in Culture

  • Malakar, Dhruba;Majumdar, A.C.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.15 no.3
    • /
    • pp.340-345
    • /
    • 2002
  • In this experiment, oocytes were collected from goat ovaries available in slaughterhouse by follicle puncture method. Morphologically culturable type of oocytes which having compact, multilayered cumulus granulosa cell complex and evenly granulated cytoplasm, was separated under a stereozoom microscope. Oocytes were washed thoroughly in maturation medium containing TCM-199, $1{\mu}g/ml$ estradiol-$17{\beta}$, 0.5 ${\mu}g/ml$ FSH, $100{\mu}g/ml$ LH, 3 mg/ml BSA and 10% estrus goat serum. Washed oocytes were cultured into maturation medium on granulosa cell monolayer. Culture plate was then kept into $CO_2$ incubator at $38{\pm}1^{\circ}C$, maximum humidity and 5% $CO_2$ for 18 h. After maturation the oocytes were washed thoroughly with maturation medium containing polyvinyl alcohol (PVA) without serum and BSA and further cultured for 12 h for secretory proteins of oocytes. PVA medium was collected, pooled and concentrated by 5000 cut off centrisart. Secretory proteins were separated on 12.5% SDS-PAGE. A total number of 3.41 oocytes per ovary were obtained and 2.17 culturable oocytes per ovary were cultured into maturation medium. After 18 h of maturation, 4,567 oocytes (1.82 oocytes per ovary) were further cultured into serum and BSA free PVA medium for its secretory proteins. Four secretory proteins of oocytes with approximately molecular weight of 45, 55, 65 and 95 kDa were obtained on SDS-PAGE in silver staining and three proteins with approximately molecular weight of 45, 55 and 65 kDa in Coomassie brilliant blue staining. In conclusion, four secretory proteins with approximately molecular weight of 45, 55, 65 and 95 kDa was obtained from in vitro cultured oocytes of goats.

Enhancement of Clavulanic Acid Production by Expressing Regulatory Genes in gap Gene Deletion Mutant of Streptomyces clavuligerus NRRL3585

  • Jnawali, Hum Nath;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
    • /
    • v.20 no.1
    • /
    • pp.146-152
    • /
    • 2010
  • Streptomyces clavuligerus NRRL3585 produces a clinically important $\beta$-lactamase inhibitor, clavulanic acid (CA). In order to increase the production of CA, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene (gap) was deleted in S. clavuligerus NRRL3585 to overcome the limited glyceraldehyde-3-phosphate pool; the replicative and integrative expressions of ccaR (specific regulator of the CA biosynthetic operon) and claR (Lys-type transcriptional activator) genes were transformed together into a deletion mutant to improve clavulanic acid production. We constructed two recombinant plasmids to enhance the production of CA in the gap1 deletion mutant of S. clavuligerus NRRL3585: pHN11 was constructed for overexpression of ccaR-claR, whereas pHN12 was constructed for their chromosomal integration. Both pHN11 and pHN12 transformants enhanced the production of CA by 2.59-fold and 5.85-fold, respectively, compared with the gap1 deletion mutant. For further enhancement of CA, we fed the pHN11 and pHN12 transformants ornithine and glycerol. Compared with the gap1 deletion mutant, ornithine increased CA production by 3.24- and 6.51-fold in the pHN11 and pHN12 transformants, respectively, glycerol increased CA by 2.96- and 6.21-fold, respectively, and ornithine and glycerol together increased CA by 3.72- and 7.02-fold, respectively.

Production of Transgenic Granulosa Cells after Retrovirus Vector Injection into Follicle in Mouse

  • Ju, Jin-Young;Chi, Hee-Jun;Koo, Jung-Jin;Kim, Teoan;Lee, Hoon-Taek;Chung, Kil-Saeng
    • Proceedings of the KSAR Conference
    • /
    • 2001.03a
    • /
    • pp.62-62
    • /
    • 2001
  • Recently, production of transgenic animal by nuclear transfer has been known as a useful method. The production of cloned offspring derived from nuclear transfer depends upon a variety of factors such as species, donor cells type and cell cycle, and source of recipient ova. Therefore, we attempted a different transgenic methods using follicular granulosa cells (GCs). In general, ovulated GCs undergoes lutenization and transformation in vitro which might defective effects on developmental potential. In order to avoid the GCs transformation in vitro culture system, we introduced a direct injection of retrovirus into the follicles and then collected them mechanically from ovaries of 6-8 week-old ICR mice. Retrovirus vector constructed with pLN $\beta$ EGFP was injected into the follicles. The follicles are cultured in $\alpha$ -MEM supplemented with human FSH, LH and ITS in Costar Transwell dish for 4 days. Survival rate of virus injected follicles was 52.1% (12/23) and expression rate of EGPP gene was 33.3% (4/12). In this study, we found GCs performed transgenesis in our culture system. In addition, the GCs in follicle may be developed in vivo like environment rather than in vitro environment. Thus, the use of GCs as donor cells may be useful in the nuclear transfer for cloning of genetic modification. Therefore, these results suggest that follicular GCs can be transfected by viral vector during folliculogenesis in vitro.

  • PDF

Endogenous catalase delays high-fat diet-induced liver injury in mice

  • Piao, Lingjuan;Choi, Jiyeon;Kwon, Guideock;Ha, Hunjoo
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.21 no.3
    • /
    • pp.317-325
    • /
    • 2017
  • Non-alcoholic fatty liver disease (NAFLD) has become the most prevalent liver disease in parallel with worldwide epidemic of obesity. Reactive oxygen species (ROS) contributes to the development and progression of NAFLD. Peroxisomes play an important role in fatty acid oxidation and ROS homeostasis, and catalase is an antioxidant exclusively expressed in peroxisome. The present study examined the role of endogenous catalase in early stage of NAFLD. 8-week-old male catalase knock-out (CKO) and age-matched C57BL/6J wild type (WT) mice were fed either a normal diet (ND: 18% of total calories from fat) or a high fat diet (HFD: 60% of total calories from fat) for 2 weeks. CKO mice gained body weight faster than WT mice at early period of HFD feeding. Plasma triglyceride and ALT, fasting plasma insulin, as well as liver lipid accumulation, inflammation (F4/80 staining), and oxidative stress (8-oxo-dG staining and nitrotyrosine level) were significantly increased in CKO but not in WT mice at 2 weeks of HFD feeding. While phosphorylation of Akt (Ser473) and $PGC1{\alpha}$ mRNA expression were decreased in both CKO and WT mice at HFD feeding, $GSK3{\beta}$ phosphorylation and Cox4-il mRNA expression in the liver were decreased only in CKO-HF mice. Taken together, the present data demonstrated that endogenous catalase exerted beneficial effects in protecting liver injury including lipid accumulation and inflammation through maintaining liver redox balance from the early stage of HFD-induced metabolic stress.

Dietary Habits and Lifestyle Factors in Relation to Sa-Sang Constitution

  • Kim Jeongseon;Kang Hye-Jung;Kim Ee-Hwa
    • Nutritional Sciences
    • /
    • v.8 no.2
    • /
    • pp.125-132
    • /
    • 2005
  • According to Sa-Sang constitution, people can be classified into 4 types, Tae-Yang-In, Tae-Eum-In, So-Yang-In, and So-Eum-In. 1he purpose of this study was to examine any significance between types of Sa-Sang constitution and dietary factors. Physical and constitutional examinations as well as a questionnaire survey were conducted on a group of 483 college students. Among the group of 483 college students, only 428 subjects were determined to have identical constitution through both the questionnaire survey and Korean traditional medical doctors' inquiries. Of 428 subjects, $29.7\%$ were determined to be Tae-Eum-Ins; $35.5\%$ were identified as So-Yang-Ins; $34.8\%$ were classified as So-Eum-Ins. Tae-Eum-Ins showed a statistically significant difference in BMI and higher smoking rate in comparison with groups of So-Yang-Ins and So-Eum-Ins. So-Eum-Ins had a tendency to take vegetables and fruits more often but they had lower preferences for balanced diets. Tae-Eum-Ins showed higher propensity to eat a visible fat protein. In terms of degrees of doneness of meat, So-Yang-Ins showed a higher tendency to prefer well-broiled meat Tae-Eum-Ins also tended to have significantly higher iron, potassium, sodium, vitamin $B_1$, niacin, $\beta$-carotene and vitamin E in their diet So-Eum-Ins had significantly lower intakes of protein, phosphorous, and folate. Considering the fact that most chronic degenerative diseases could be developed by any lifestyle factors, it is necessary to conduct educational programs about lifestyles including dietary habits for maintaining good health; On the basis of the results of this study, it is expected that the scientific, objective and accurate diet information depending upon individual's type of constitution will be provided.

Liripois Tuber contributes to the chemotaxis of eosinophils and secretion of cytokines in A549 human epithelial cells (맥문동(麥門冬)이 천식유발 cytokine 분비와 호산구 chemotaxis에 미치는 영향)

  • Jung, Hae-Jun;Jung, Gee-Jae;Jung, Sung-Ki;Rhee, Hyung-Koo
    • The Journal of Korean Oriental Chronic Disease
    • /
    • v.10 no.1
    • /
    • pp.1-20
    • /
    • 2005
  • Background and Objective: The importance of the presence of eosinophils in the airways of patients with fetal asthma has long been recognized, but the mechanism by which these cells are recruited and retained in the lung are only now being elucidated. Eotaxin is a potent and specific eosinophil chemoattractant that is mobilized in the respiratory epithelium after allergic stimulation. Material and Methods: We used water extracts of Liripois Tuber and pulmonary epithelial cell lines A549(human typeII-like epithelial cells) and human eosinophils. We estimated cytotoxic effects of Liripois Tuber via MTS assay, and estimated the effects of Liripois Tuber on chemokines from prestimulated A549 cells by sandwich ELISA and RT-PCR. We conducted chemotaxis assay on prestimulated eosinophils treated with Liripois Tuber. Result: In this study we demonstrated that $TNF-{\alpha}$, IL-4 and $IL-1{\beta}$ induced the accumulation of chemokines mRNA in the pulmonary epithelial cell lines A549 in dose-dependent manner. Chemokines were inhibited by Liripois Tuber in dose-dependent manner. The eosinophil migration was inhibited at high concentration of Liripois Tuber. Conculusion: These findings indicate that the supression of the expression of chemokines can be accomplished by Liripois Tuber treatment, raising the possibility that Liripois Tuber might be of therapeutic value in diseases such as asthma.

  • PDF

Affecting Factors in Job-seeking Stress among Health Care Related Students (일 지역 보건의료계열 대학생의 취업스트레스 영향요인)

  • Bang, Miran;Sim, Sunsook
    • The Journal of the Korea Contents Association
    • /
    • v.18 no.10
    • /
    • pp.257-266
    • /
    • 2018
  • The purpose of this study is to investigate the relationship between self-esteem, appearance satisfaction, interpersonal relationship, and job-seeking stress in health care related students and to investigate the factors influencing college students' job-seeking stress. A self-report type survey was conducted targeting a total of 184 of college students in a city from June 11 to 15, 2018 were enrolled. The results are as followed; The strongest predictor of job-seeking stress was self-esteem (${\beta}=-.407$, p<.001). Self-esteem, employment impact of appearance, academic grade accounted for 18.5% of job-seeking stress based on multiple regression analysis. In order to reduce the job-seeking stress of health care related students, it is necessary to develop a personalized employment program for each major field and individual that can improve the self-esteem of the individual.

New metabolites from the biotransformation of ginsenoside Rb1 by Paecilomyces bainier sp.229 and activities in inducing osteogenic differentiation by Wnt/β-catenin signaling activation

  • Zhou, Wei;Huang, Hai;Zhu, Haiyan;Zhou, Pei;Shi, Xunlong
    • Journal of Ginseng Research
    • /
    • v.42 no.2
    • /
    • pp.199-207
    • /
    • 2018
  • Background: Ginseng is a well-known traditional Chinese medicine that has been widely used in a range of therapeutic and healthcare applications in East Asian countries. Microbial transformation is regarded as an effective and useful technology in modification of nature products for finding new chemical derivatives with potent bioactivities. In this study, three minor derivatives of ginsenoside compound K were isolated and the inducing effects in the Wingless-type MMTV integration site (Wnt) signaling pathway were also investigated. Methods: New compounds were purified from scale-up fermentation of ginsenoside Rb1 by Paecilomyces bainier sp. 229 through repeated silica gel column chromatography and high pressure liquid chromatography. Their structures were determined based on spectral data and X-ray diffraction. The inductive activities of these compounds on the Wnt signaling pathway were conducted on MC3T3-E1 cells by quantitative real-time polymerase chain reaction analysis. Results: The structures of a known 3-keto derivative and two new dehydrogenated metabolites were elucidated. The crystal structure of the 3-keto derivative was reported for the first time and its conformation was compared with that of ginsenoside compound K. The inductive effects of these compounds on osteogenic differentiation by activating the Wnt/b-catenin signaling pathway were explained for the first time. Conclusion: This study may provide a new insight into the metabolic pathway of ginsenoside by microbial transformation. In addition, the results might provide a reasonable explanation for the activity of ginseng in treating osteoporosis and supply good monomer ginsenoside resources for nutraceutical or pharmaceutical development.

Wollastonite from and Its Dissolution Behaviors (수산 지역의 규회석과 그 용해 거동)

  • 김수진;현성필;이성근
    • Journal of the Mineralogical Society of Korea
    • /
    • v.9 no.1
    • /
    • pp.1-6
    • /
    • 1996
  • Woolastonite from Susan occurs as intercalations in limestone beds of Lower Paleozoic Joseon Supergroup. It is a thermal metamorphic product of impure limestone. Electron microprobe analysis shows that it is considerably pure wollastonite. It has triclinic cell with a=7.932$\AA$, b=7.328$\AA$, c=7.069$\AA$, $\alpha$=89.995$^{\circ}$, $\beta$=$95.255^{\circ}$, and $ \Upsilon=103.367^{\circ}$.Dissolution behaviors of wollastonite have been studied conducting three different dissolution experiments; two different reactions with HC1 (one batch and one re-initialization experiment) and one traction with distilled water. In the batch type powder wollastonite-HCl reaction, pH of solution rapidly increases in the early stage and then its rate of increase slows down to reach plateau resulting in parabolic relationship with time. It is represented by the early rapid rise and fall in pH giving a sharp pH-edge and succeeding slow rise in the re-initialization experiment. The early rapid rise in pH is due to the rapid sorption of H- in solution to oxygens on the reactive surface of wollastonite and the fall in pH means that all reactive surface sites are occupied by H- ions and no more H- adsorption occurs. The slow rise in pH following the pH- edge is due to the dissolution of wollastonite as evidenced by the correlation of pH variation and cation concentration. Dissolution of powder wollastonite in HCl shows linear trend with time. Si is dissolved predominantly over Ca at a constant rate. Ca is dissolved predominantly in the very early stage. Dissolution rate of coarse-grained wollastonite fragments in distilled water is parabolic with times howing a rapid reaction in the early stage and a slow reaction in the advanced stage. The Ca/Si ratio in solution is high in the case of coarse-grained wollastonite fragment as compared with powder wollastonite. The coarse-grained wollastonite fragment-water (acid) reaction resulted in the solution with an elevated constant pH value (alkaline) giving an important significance on the environmental view point.

  • PDF

Fabrication of ACtA/$SiC_w$ composite by squeeze casting (I) (용탕 단조법에 의한 AC4A/Si$C_w$복합재료 제조에 관한 연구 (I))

  • Moon, Kyung-Cheol;Lee, Jun-Hee
    • Korean Journal of Materials Research
    • /
    • v.2 no.6
    • /
    • pp.461-467
    • /
    • 1992
  • A fabrication process for SiC whisker preform reinforced AC4A Al composites is being developed. The Al alloy used as the matrix in this study is AC4A. SiC whisker preform made by Tokai Carbon Co. Ltd. Shizuoka, Japan were used. These consisted of $\beta$-type single crystals 0.1 ~ 10${\mu}$m in diameter and 20~10${\mu}$m in length. The most adequate fabrication condition was that whisker preform was preheated up to 750~80$0^{\circ}C$, set into a mould preheated to ~40$0^{\circ}C$, molten Al alloy heated to ~80$0^{\circ}C$ and applied pressure 75MPa. And Si$C_w$reinforced AC4A composite was advanced above twice than AC4AI/M. Also it was not large effect by pressure at Si$C_w$ 20v/o.

  • PDF