• Title/Summary/Keyword: germ activity

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Migration Activity of Chicken Gonadal Primordial Germ Cells (gPGCs) and Post-transfer Localization of LacZ-transfected gPGCs in the Embryonic Gonads

  • Jeong, D.K.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1227-1231
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    • 2002
  • A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.

Phytosterols content of Keunnunjami germ and its antioxidative effects in adult rats (큰눈자미 배아의 식물성 스테롤 함량 및 성숙기 흰쥐에서 항산화 효과)

  • Liang, Jie;Ma, Jing Wen;Chung, Soo Im;Kang, Mi Young
    • Journal of Nutrition and Health
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    • v.53 no.2
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    • pp.99-110
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    • 2020
  • Purpose: The rice germ fraction is a better source of protein, lipid, and fiber than the rice endosperm. Furthermore, the rice germ is rich in bioactive phytochemicals such as γ-aminobutyric acid, tocopherols, tocotrienols, phytic acid, and so on. In this study, the phytosterol content and antioxidant activity of Keunnunjami germ (KG) or normal rice germ supplement were investigated in healthy adult rats. Methods: In vitro, quantitative assessment of phytosterols, including β-sitosterol, campesterol, cycloartenol, and stigmasterol, was performed. Comparative antioxidant activities of 2 rice germs were measured based on DPPH radical scavenging activity, reducing power, and ABTS radical scavenging capacity. In vivo, male Spraque-Dawley rats (30-weeks-old) were randomly assigned a diet of normal control (NC, AIN-93M diet), AIN-93M diet supplemented with normal rice germ 3% (NG3), or AIN-93M diet supplemented with KG 3% (KG3) and fed for 8 weeks. Results: KG contained significantly higher campesterol and stigmasterol contents and antioxidant activity than normal rice germ. The KG3 group exhibited significantly lower body weight gain as well as inguinal and total white adipose tissue weights. There were no significant differences in plasma glucose, insulin, C-peptide, or homeostasis model assessment of insulin resistance level among the 3 groups. The plasma tumor necrosis factor-α concentration was significantly lower while leptin, advanced oxidation protein products, and interleukin-6 showed downward trends in the KG3 group. In addition, the superoxide dismutase level of the KG3 group was significantly higher compared to the NC and NG3 groups. Conclusion: This study indicates that KG can be considered as a valuable source of phytosterol components. Lastly, KG has strong antioxidant properties and may have potential to ameliorate elevation of proinflammatory cytokine production with age.

Relationship between Germ Tube Formation, Adherence to Human Buccal Epithelial Cells and Virulence of Candida albicans (Candida albicans의 상피세포에 대한 부착능과 병원성과의 상관관계에 관한 연구)

  • Koh, Choon-Myung
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.407-415
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    • 1986
  • This study investigated whether a correlation exists between environmental physical and biochemical factors and adherence of Candida albicans to human buccal epithelial cells by using normal and UV-irradiated strains. The results were as follows: 1. The percentage of germ tube forming activities of normal Candida albicans was 91.5% and UV-irradiated Candida albicans was 15.0%. The $LD_{50}$ of normal strains in mice were $1.0{\times}10\;cells/ml$, but could not be observed in the UV-irradiated strains even with $1.0{\times}10\;cells/ml$. It demonstrated that the virulence is decreased in the UV-irradiated strain. 2. The adherence of normal Candida albicans to human buccal epithelial cells($166{\pm}29{\sim}207{\pm}17\;cells$/100 epithelial cells) was significantly greater than UV-irradiated Candida albicans($99{\pm}21{\sim}131{\pm}25\;cells$/100 epithelial cells). 3. Candida albicans cultured at $37^{\circ}C$ adhered to buccal epithelial cells($166{\pm}16{\sim}207{\pm}17\;cells$/100 epithelial cells) in greater numbers than cultured at $25^{\circ}C$($80{\pm}15{\sim}143{\pm}22\;cells$/100 epithelial cells). 4. On comparison of the adherence of viable and nonviable(heat-killed) Candida albicans to human buccal epithelial cells, the nonviable Candida albicans demonstrated poorer adherence than viable Candida albicans. 5. Adherence in vitro of Candida albicans to human epithelial cells appeared to be effected by the pH. The adherence ability was maximum increased at pH 7.0($187{\pm}22\;cells$/100 epithelial cells) other than experimental pH. 6. The adherence was proportional to the incubation time and the Candida cell concentration in the suspension. 7. A strong correlation was shown between germ tube forming activity and increased adherence of Candida albicans to human epithelial cells, indicating that germ tube forming activity were responsible for candidal virulence.

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Identification and Characterization of Rodent Germ Cells-Specific Hyaluronidases

  • Kim, Ekyune;Chang, Kyu-Tae
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.155-161
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    • 2012
  • Germ cell-specific hyaluronidases such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5) are in part responsible for dispersal of the cumulus cell mass, which is a critical step in establishing fertilization in mammals. In this study, we identified two testis-hyaluronidases, SPAM1 and Hyal5, in hamster and rat. These two genes were expressed specifically in the testis. At the protein level, hamster SPAM1 and Hyal5 display 78.7% and 75.4% identity with mouse SPAM1 and Hyal5. Further, the activity of the enzymes with respect to cumulus cell dispersion did not differ, although we observed that the enzymatic activity differed in pH range. These studies suggest that different sperm hyaluronidases are capable of dispersing the cumulus cell mass despite differences in enzyme activity.

A Mutation of cdc-25.1 Causes Defects in Germ Cells But Not in Somatic Tissues in C. elegans

  • Kim, Jiyoung;Lee, Ah-Reum;Kawasaki, Ichiro;Strome, Susan;Shim, Yhong-Hee
    • Molecules and Cells
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    • v.28 no.1
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    • pp.43-48
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    • 2009
  • By screening C. elegans mutants for severe defects in germline proliferation, we isolated a new loss-of-function allele of cdc-25.1, bn115. bn115 and another previously identified loss-of-function allele nr2036 do not exhibit noticeable cell division defects in the somatic tissues but have reduced numbers of germ cells and are sterile, indicating that cdc-25.1 functions predominantly in the germ line during postembryonic development, and that cdc-25.1 activity is probably not required in somatic lineages during larval development. We analyzed cell division of germ cells and somatic tissues in bn115 homozygotes with germline-specific anti-PGL-1 immunofluorescence and GFP transgenes that express in intestinal cells, in distal tip cells, and in gonadal sheath cells, respectively. We also analyzed the expression pattern of cdc-25.1 with conventional and quantitative RT-PCR. In the presence of three other family members of cdc-25.1 in C. elegans, defects are observed only in the germ line but not in the somatic tissues in cdc-25.1 single mutants, and cdc-25.1 is expressed predominantly, if not exclusively, in the germ line during postembryonic stages. Our findings indicate that the function of cdc-25.1 is unique in the germ line but likely redundant with other members in the soma.

Depletion Effects of Chick Germinal Crescent's Primordial Germ Cells by Heat Activated Busulfan Injection (닭 생식반월의 Busulfan 가온 주입방법에 의한 원시생식세포 제거 효과)

  • Jeong, Dong-Kee
    • Development and Reproduction
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    • v.11 no.3
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    • pp.219-226
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    • 2007
  • This study was conducted to identify optimistic primordial germ cells'(PGCs) migration activity using heat activated busulfan treatment for the increasing germline chimerism. Donar PGCs viability tests of important conditions for useful germ line chimerism indicated approximately $70{\sim}80%$ viability was time dependent. Transplantation experiments of PGCs into recipient embryos after busulfun treatment, showed the treatment group having 23.5% viability. By comparison, the control group showed 4.8% viability. The 96 hour treatment group and the 118 hour treatment group of the cultured PGCs showed high migration activity. Generally, the transplantation method would consider morphological and physiological characteristics before transplantation. In the present study, the effect of busulfan on migration activity showed viability highest at 53.4% after 48-hour incubation time. However, a previous study showed the best condition for transplantation time to be prior to the 48-hour incubation period, when the chicken embryo does not yet have a developed blood vessel system. In conclusion, an important condition for the production of a transgenic chicken is that most donor PGCs migrate into the recipient embryo without any inhibitory factors. The present results suggest, perhaps by using this modified method of transplantation, it can produce a more efficient chimeric germ line, transgenic chicken.

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Infertility of Transgenic Mice Experssing Human Growth Hormone Gene (사람 성장호르몬 유전자를 발현하는 형질전환생쥐의 불임성)

  • 한용만;강만종;이철상;유대열;이경광
    • Korean Journal of Animal Reproduction
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    • v.16 no.3
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    • pp.225-230
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    • 1992
  • Many transgenic mice expressing human growth hormone gene were infertile. To investigate the infertility of these transfenic mice, it was looked into the estrus cycle and sexual behaviour and also tested through in vitro fertilization whether the germ cells of these mice normal or not. The infertile female transgenic mice were mated to the fertile males of ICR strain, but in almost all of them the vaginal plugs were not detected and their estrus cycles by vaginal smear were almost irregular which kept up estrus or diestrus stage. Many male transgenic mice did not have the ability of sexual behaviour. Therefore the viability of germ cells in infertile male transgenic mice was investigated by in vitro fertilization, but the sperm were normally fertilized with the eggs and the transgene of parent was passed on to the progeny. These results consequently suggest that the infertility of transgenic mice experssing human growth hormone gene may be due to the physiological activity of human growth hormone, not germ cells.

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Antioxidative Activity of Barley Polyphenol Extract (BPE) Separated from Pearling By-products (도정부산물로부터 분리한 보리 폴리페놀 추출물의 항산화 효과)

  • Seog, Ho-Moon;Seo, Mi-Sook;Kim, Heung-Man;Ahn, Myung-Soo;Lee, Young-Tack
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.889-892
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    • 2002
  • Barley bran and germ fractions were collected during pearling process. Barley polyphenol extract (BPE) was separated from the pearling fractions, and its antioxidative activity was determined through linoleic acid model system and lipid autoxidation model using corn oil as a substrate. At 0.02% addition level, thiobarbituric acid (TBA) value of BPE from bran fraction III appeared to be similar to that of ${\alpha}-tocopherol$, and bran fraction I and germ extract exhibited similar antioxidative activities to that of BHT. Peroxide value was measured to estimate antioxidative activity of BPE upon lipid autoxidation. As BPE concentration increased, higher antioxidative activity was observed compared to ${\alpha}-tocopherol$ and BHT until 10-15 day storage, whereas no effect was shown after 20-day storage. Relative antioxidant effectiveness (RAE) of $0.02{\sim}0.1%$ BPE from bran fraction I and germ were $128{\sim}135$ and $126{\sim}133$, respectively, and appeared to be higher than that of BHT (126), suggesting that these BPE fractions could be used as natural antioxidants.

Lipoxygenase and Off-flavor Development in Some Frozen Foods (일부냉동식품에서의 Lipoxygenase와 이취발생관계)

  • Lee, Young-Chun
    • Korean Journal of Food Science and Technology
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    • v.13 no.1
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    • pp.53-56
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    • 1981
  • Several tests were conducted to study lipoxygenase activity and off-flavor developement in frozen sweet corn. Fresh corn contained about 60% of total lipoxygenase activity in the germ section. When non-blanched frozen sweet corn was stored at $-10^{\circ}F$, it developed off-flavor and most significant changes in the flavor profile of off-flavored sweet corn was $4{\sim}5$ times higher hexanal peaks. The high hexanal peaks observed in the sterilized sweet corn with added lipoxygenase, alone and in combination with other enzymes, suggested the fact that high hexanal peaks in off-flavored sweet corn could be due to an oxidative reaction of lionleic acid (and other unsaturated fatty acids) catalyzed by lipoxygenase. Based on lipoxygenase activity and linoleic acid content in sweet corn, this reaction occur most heavily in the germ section of sweet corn. There was a significant relationship between flavor score of frozen stored corn-on-the-cob and hexanal peak in the germ section of corn-on-the-cob. This result indicated that hexanal peak could be used as an objective index of off-flavor development in frozen sweet corn.

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Anti-Oomycete Activity and Pepper Root Colonization of Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 against Phytophthora capsici

  • Elena, Volynchikova;Ki Deok, Kim
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.123-135
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    • 2023
  • Previously, Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 from a sequential screening procedure were proven to effectively control Phytophthora blight caused by Phytophthora capsici. In this study, we further investigated the anti-oomycete activities of these strains against mycelial growth, zoospore germination, and germ tube elongation of P. capsici. We also investigated root colonization ability of the bacterial strains in square dishes, including cell motility (swimming and swarming motilities) and biofilm formation. Both strains significantly inhibited mycelial growth in liquid and solid V8 juice media and M9 minimal media, zoospore germination, and germ tube elongation compared with Bacillus vallismortis EXTN-1 (positive biocontrol strain), Sphingomonas aquatilis KU408 (negative biocontrol strain), and MgSO4 solution (untreated control). In diluted (nutrient-deficient) V8 juice broth, the tested strain populations were maintained at >108 cells/ml, simultaneously providing mycelial inhibitory activity. Additionally, these strains colonized pepper roots at a 106 cells/ml concentration for 7 days. The root colonization of the strains was supported by strong swimming and swarming activities, biofilm formation, and chemotactic activity towards exudate components (amino acids, organic acids, and sugars) of pepper roots. Collectively, these results suggest that strains YJR13 and YJR92 can effectively suppress Phytophthora blight of pepper through direct anti-oomycete activities against mycelial growth, zoospore germination and germ tube elongation. Bacterial colonization of pepper roots may be mediated by cell motility and biofilm formation together with chemotaxis to root exudates.