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Effects of intrauterine growth restriction during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses

  • Zi, Yang;Ma, Chi;He, Shan;Yang, Huan;Zhang, Min;Gao, Feng;Liu, Yingchun
    • Animal Bioscience
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    • v.35 no.7
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    • pp.989-998
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    • 2022
  • Objective: This study investigated the effects of intrauterine growth restriction (IUGR) during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses. Methods: Eighteen time-mated Mongolian ewes with singleton fetuses were allocated to three groups at d 90 of pregnancy: restricted group 1 (RG1, 0.18 MJ ME/body weight [BW]0.75/d, n = 6), restricted group 2 (RG2, 0.33 MJ ME/BW0.75/d, n = 6) and control group (CG, ad libitum, 0.67 MJ ME/BW0.75/d, n = 6). Fetuses were recovered at slaughter on d 140. Results: The G0/G1 phase cell number in fetal thymus of the RG1 group was increased but the proliferation index and the expression of proliferating cell nuclear antigen (PCNA) were reduced compared with the CG group (p<0.05). Fetuses in the RG1 group exhibited decreased growth hormone receptor (GHR), insulin-like growth factor 2 receptor (IGF-2R), and their mRNA expressions (p<0.05). For the RG2 fetuses, there were no differences in the proliferation index and PCNA expression (p>0.05), but growth hormone (GH) and the mRNA expression of GHR were lower than those of the CG group (p<0.05). The thymic mRNA expressions of cyclin-dependent protein kinases (CDKs including CDK1, CDK2, and CDK4), CCNE, E2-factors (E2F1, E2F2, and E2F5) were reduced in the RG1 and RG2 groups (p<0.05), and decreased mRNA expressions of E2F4, CCNA, CCNB, and CCND were occurred in the RG1 fetuses (p<0.05). The decreased E-cadherin (E-cad) as a marker for epithelial-mesenchymal transition (EMT) was found in the RG1 and RG2 groups (p<0.05), but the OB-cadherin which is a marker for activated fibroblasts was increased in fetal thymus of the RG1 group (p<0.05). Conclusion: These results indicate that weakened GH/IGF signaling system repressed the cell cycle progression in G0/G1 phase in IUGR fetal thymus, but the switch from reduced E-cad to increased OB-cadherin suggests that transdifferentiation process of EMT associated with fibrogenesis was strengthened. The impaired cell growth, retarded proliferation and modified differentiation were responsible for impaired maturation of IUGR fetal thymus.

The Analysis of Dissociation Properties According to Gas Hydrate Saturation and Depressurization Rate (가스하이드레이트 포화율 및 감압률에 따른 해리특성 분석)

  • An, Seung-Hee;Chon, Bo-Hyun
    • Journal of the Korean Institute of Gas
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    • v.19 no.3
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    • pp.54-59
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    • 2015
  • The gas hydrate of 10 trillion tons are buried under continental slope in the world(permafrost : 2%, marine continental slope: 98%), but technology for the the commercial gas recovery has not developed yet. There are normally four representative recovery methods: depressurization method, thermal stimulation method, inhibition injection method, and displacement method. This study focuses on change of dissociation time and gas production according to gas hydrate saturation rate and depressurization rate. It was found that the correlation between depressrization rate and dissociation time was like as $Y=0.0004X^2-0.499X+176.86$. It was also found that the bigger depressurization rate is, the better production is(methane gas is produced over 46.2% at depressurization rate 50% compared with 40%). However, on the contrary to this, it is presumed that gas production is decreased at 60% due to gas hydrate reformation.

DdeI Polymorphism in Coding Region of Goat POU1F1 Gene and Its Association with Production Traits

  • Lan, X.Y.;Pan, C.Y.;Chen, H.;Lei, C.Z.;Hua, L.S.;Yang, X.B.;Qiu, G.Y.;Zhang, R.F.;Lun, Y.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.9
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    • pp.1342-1348
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    • 2007
  • POU1F1 is a positive regulator for GH, PRL and TSH${\beta}$ and its mutations associate with production traits in ruminant animals. We described a DdeI PCR-RFLP method for detecting a silent allele in the goat POU1F1 gene: TCT (241Ser)>TCG (241Ser). Frequencies of $D_1$ allele varied from 0.600 to 1.000 in Chinese 801 goats. Significant associations of DdeI polymorphism with production traits were found in milk yield (*p<0.05), litter size (*p<0.05) and one-year-old weight (*p<0.05) between different genotypes. Individuals with genotype $D_1D_1$ had a superior performances when compared to those with genotype $D_1D_2$ (*p<0.05). Hence, the POU1F1 gene was suggested to the potential candidate gene for superior milk performance, reproduction trait and weight trait. Genotype $D_1D_1$, characterized by a DdeI PCR-RFLP detection, was recommended to geneticists and breeders as a molecular marker for better performance in the goat industry.

Cotreatment with Growth Hormone in Controlled Ovarian Hyperstimulation for IVF in Women with Limited Ovarian Reserve (체외수정시술을 위한 성선자극호르몬 과배란유도에 Limited Ovarian Reserve를 갖는 환자에서 성장호르몬의 사용)

  • Kim, Sun-Haeng;Chang, Ki-Hoon;Ku, Pyoung-Sahm
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.3
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    • pp.241-245
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    • 1994
  • Despite increasing success rate of IVF, poor response to ovarian stimulation remains a problem. So, attempts to improve ovarian responses, for example, by using combined gonadotropin-releasing hormone analogue(GnRH-a) and human menopausal gonadotropin(hMG) have shown limited success. It is reported that response of granulosa cells in vitro to FSH is stimulated by co-incubation with IGF-l, and IGF-l production can be increased by growth hormone. This suggest that combination regimen of G.H. and hMG may augment follicle recruitment. In fifteen patients who had previous history of poor ovarian response to gonadotropin stimulation after pituitary suppression with mid -luteal GnRH-a, the effectiveness of cotreatment with G.H. in IVF program was evaluated using a combination regimen of G.R. and hMG at Korea University Hospital IVF Clinic. Ovarian responses to gonadotropin stimulation in control and GH-treated cycles assessed by total dose and duration of hMG treatment, follicular development and peak $E_2$ level, number of eggs retrieved, and fertilization rates were also assessed. In each group, serum and follicular fluid IGF-1 concentrations on day of egg collection were measured by RIA after acidification and extraction by reveresed phase chromatography. Patients receiving G.H. required fewer days and ampules of gonadotropins, developed more oocytes, and more embryos transferred. But, the differences were not statistically significant, except the duration of hMG treatment. Our data showed a significantly higher concentration of IGF-l in the serum, not in the follicular fluid, of patients treated with G.H. compared with control group. These data suggest that growth hormone treatment does not improve the ovarian response in women with limited ovarian reserve to gonadotropin stimulation for IVF.

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Isolation and characterization of a novel gossypol-degrading bacteria Bacillus subtilis strain Rumen Bacillus Subtilis

  • Zhang, Yunhua;Zhang, Zhengyou;Dai, Li;Liu, Ying;Cheng, Maoji;Chen, Lijuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.63-70
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    • 2018
  • Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

Isolation of Mannanase-producing Bacteria, Bacillus subtilis WL-6 and WL-11, and Cloning and Characterization of Mannanase (Bacillus subtilis 분리균 2주 유래 mannanases의 특성 비교)

  • Yoon, Ki-Hong
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1113-1120
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    • 2016
  • Two bacterial strains producing extracellular man nanase were isolated from doenjang, a traditionally fermented soybean paste in Korea. The isolates, WL-6 and WL-11, were identified as Bacillus subtiis on the basis of their 16S rRNA gene sequences, morphological, and biochemical properties. Two genes encoding the mannanase of both B. subtilis WL-6 and B. subtilis WL-11 were each cloned into Escherichia coli, and their nucleotide sequences were determined. Both mannanase genes consisted of 1,086 nucleotides, encoding polypeptides of 362 amino acid residues. The deduced amino acid sequences of the two WL-6 and WL-11 mannanases, designated Man6 and Man11, respectively, differed from each other by eight amino acid residues, and they were highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The 26 amino acid stretch in the N-terminus of Man6 and Man11 was a predicted signal peptide. Both Man6 and Man11 were localized at the level of 94–95% in an intracellular fraction of recombinant E. coli cells. The enzymes hydrolyzed both locust bean gum and mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, forming mannobiose and mannotriose as predominant products. The optimal reaction conditions were 55°C and pH 6.0 for Man6, and 60°C and pH 5.5 for Man11. Man11 was more stable than Man6 at high temperatures.

Immobilization of $\beta-galactosidase$ with High Transgalactosylation Activity from Bacillus sp. A4442 Mutant (변이주 Bacillus sp. A4442가 생산하는 갈락토스 전이활성이 높은 $\beta-galactosidase$의 고정화)

  • Kim, Min-Hong;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.333-337
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    • 1996
  • For continuous production of galactooligosaccharides(GOS), $\beta-galactosidase$ with h1gh transgalactosylation activity from Bacillus sp. A 4442 was Immobilized onto $Diaion^{TM}$ HPA 75(styrene-divinylbenzene resin). The parameters influencing enzyme immobilization were scrutinized in order to maximize immobilization yield while minimizing enzyme inactivation. The optimum conditions turned out to be: Tris buffer concentration 30 mM, pH 8.0, contact time at room temperature 3 hr, and enzyme loading 25 mg protein/g resin. Both the thermal stability and the operational stability of immobilized enzyme were markedly enchanced by the treatment with 0.5% glutaraldehyde as a cross-linker. Under the experimental conditions established, the yield of ${\beta}-galactosidase$ immobilization was 40% or more and the activity of the immobilized enzyme ca. 200 U/g resin. When a packed-bed reactor was employed to continuously convert lactose to GOS, the specific production, which refers to as the amount of commercially valuable GOS produced by a unit amount of immobilized ${\beta}-galactosidase$, was found to be ca. 300 g GOS/g carrier.

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The Estrous Cycle of the Markhoz Goat in Iran

  • Farshad, A.;Akhondzadeh, M.J.;Zamiri, M.J.;Sadeghi, GH.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1411-1415
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    • 2008
  • The purpose of the present study was to determine the onset of breeding season, the occurrence of silent and true heats and the duration of estrus in female Markhoz goats of the Kurdistan province in Iran. Ten, 3 years-old Markhoz does with an average weight of $34.05{\pm}2.62kg$ and with one kidding record, were used. The goats were maintained in an open barn under constant nutritional levels and natural photoperiod. One aproned buck was used twice a day every 12 h to detect estrus from mid August to early January. For the determination of the onset of reproductive activity as well as occurrence of silent heat, blood samples were collected every 10 days, from the beginning of the experiment. After $2^{nd}$ standing heat, blood samples were obtained twice a week in order to assess luteal activity and the length of estrous cycles. In this study, estrous behavior was observed including sniffing, vocal exchange, following courtship, flehman, standing heat and mounting. The results of the progesterone assay indicated that in this goat silent heats occur mostly in the early breeding season. The first standing heat was observed in mid-October which was considered as the onset of the breeding season. Duration of the estrous cycle and estrus was recorded as being $20.93{\pm}1.56days$ and $38.86{\pm}15.19h$, respectively. The correlation coefficient between length of daylight and occurrence of estrus was negative (r = -0.470) but not significant (p>0.05). The data showed that there was no significant effect of body weight on estrous cycles (first, second and third) and estrous periods (first, second, third and fourth). Progesterone levels were not significantly different in the first, second and third estrous cycles at days 0, 4, 10, 12, 14 and 19. The results of progesterone assay during the estrous cycle indicate that follicular and luteal phases last 4-5 and 14-15 days, respectively and the concentration of serum progesterone in these phases was $0.88{\pm}0.08$ and $7.44{\pm}0.26ng/ml$, respectively. The study concluded that Markhoz does could be considered as a breed with a short breeding season and an optimal estrous activity in autumn.

Somatic Embryogenesis in Immature Zygotic Embryo Cultures of Korean Soybean (Glycine max L.) Culitivars and Effect of 2,4-Dichlorophenoxyacetic Acid on Somatic Embryo Morphology (한국 품종 대두(Glycine max L) 미숙배로부터 체세포배발생과 배지의 2,4-Dichlorophenoxyacetic Acid 농도가 체세포배의 형태에 미치는 영향)

  • Pil S Choi;Yoong Y. SOH;Duck Y.Choi;ang R. LIU
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.7-14
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    • 1994
  • Immature zygotic embryos of five Korean soybean cultivars cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) produced somatic embryos without forming an intervening callus. The highest frequency (up to 90%) of somatic embryo formation was obtained when zygotic embryos were cultlued on medium containing 1 to 2 mga 2, 0-D in four cultivars. However the frequency was highly variable to the cultivars. Transversely sliced primary somatic embryo halves were also capable of forming secondary embryos at frequencies of up to 70% when cultured on medium containing 0.1 to 1 mg/L 2,4-D. Somatic embryos formed on zygotic embryos cultured on medium containing 0.1 to 0.2 mg/L 2,4-D had two cotyledons more frequently than one horn-type cotyledon and those on medium containing 0.5 to 4mg/L 2,4-LD had a horm-type cotyledon at a prominently higher freequency. However somatic embryos on medium containing 10mg/L or higher concentrations of 2,4-D were usually shunted at the globular stage even after transfer to medium containing lower concentrations of 2,4-D or other growth regulators. non somatic embryos with one or two cotyledons or a hem-type cotyledon were transferred to medium containing $GA_3$, those with two cotyledons converted to plantlets at a higher frequency (25%) than the others.

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Serum levels of free insulin-like growth factor-I and clinical value in healthy children (한국 소아 및 청소년에서 혈중 유리 insulin-like growth factor-I 농도치와 임상적 의의)

  • Chung, Young Hee;Chung, Woo Yeong
    • Clinical and Experimental Pediatrics
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    • v.51 no.1
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    • pp.47-53
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    • 2008
  • Purpose : The serum levels of total insulin-like growth factor (IGF)-I and IGF binding protein (IGFBP)-3 reflect endogenous growth hormone (GH) secretion in healthy children. Free form of IGF-I which is suggested to have more potent biological action than complex form of IGF-I. The aim of this study is to investigate the serum levels of free IGF-I and its clinical value in healthy children. Methods : Serum levels of total IGF-I and IGFBP-3 were determined in 494 healthy children (248 boys and 246 girls) by RIA and IRMA. Serum level of free IGF-I was determined in 206 healthy children (103 boys and 103 girls) by IRMA. Results : The free IGF-I level increased with age in both sex. The free IGF-I level increased continuously between 7 and 15 years of age in boys, but decrement was noted after 14 years of age in girls. Serum total IGF-I level also increased with age in similar pattern of that of free IGF-I. There were no significant differences of mean values of the ratio of free IGF-I/total IGF-I in relation to age in both sex. And there were significant correlations between the level of free IGF-I and total IGF-I and the ratio of total IGF-I/IGFBP-3, respectively. Conclusion : In healthy children, serum free IGF-I increased with age in both sex and high free IGF-I level may play an important role in pubertal growth spurt. Our results suggest that the increased serum free IGF-I level in puberty may reflect changes in total IGF-I rather than IGFBP-3. But free IGF-I does not have more clinical value than total IGF-I because of no significant differences of mean values of the ratio of free IGF-I/total IGF-I in relation to age.