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The Effect of Sonicated Extracts of Treponema Denticola and Treponema Lecithinolyticum on the Cytokine Secretion and Matrix Metalloproteinase Activation of Gingival Fibroblast (Treponema denticola와 Treponema lecithinolyticum의 분쇄액이 치은섬유아세포의 Cytokine 분비 및 Matrix metalloproteinase 활성에 미치는 영향)

  • Suh, Hye-Yuhn;Choi, Bong-Kyu;Choi, Seong-Ho;Cho, Kyoo-Sung;Kim, Chong-Kwan;Chai, Jung-Kiu
    • Journal of Periodontal and Implant Science
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    • v.29 no.4
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    • pp.979-995
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    • 1999
  • This study was investigated to observe the effect of Treponema denticola cell sonicates(TDC) and Treponema lecithinolyticum cell sonicates(TLC) on cytokine secretion and matix metalloproteinase-2(MMP-2) activation of cultured human gingival fibroblast. Several experiments were performed including $IL-1{\beta}$, IL-6 ELISA for the effect on the $IL-1{\beta}$, IL-6 secretion of human gingival fibroblast. Also gelatinase zymography and gelatin dissolubility test for the activation of MMP-2 secreted by gingival fibroblast. The results were as follows. 1. The effect of TDC and TLC on IL-6 secretion of human gingival fibroblast showed statistically significant increase of IL-6 secretion in the TDC and TLC treated group compared to no treatment group(p<0.05) . 2. The amount of $IL-1{\beta}$ secretion was below the lower limit and there was no difference in the $IL-1{\beta}$ secretion of gingival fibroblast between TDC, TLC treated group and no treatment group. 3. The active form of pro MMP-2 with 72 kDa molecular weight was activated in both TDC and TLC treated group and clear band was appeared at 62kDa site on the zymography. 4. Gelatin dissolubility of MMP-2 secreted by gingival fibroblast was higher in TDC and TLC treated group compared to no treatment group(p<0.05). 5. In the TDC treated group, serine protease of T. denticola affect gelatin dissolubility. But in the TLC treated group gelatin was degraded by only MMP secreted by gingival fibroblast. Regarding to the above results, TDC and TLC have an effect on the IL-6 secretion increase of human gingival fibroblast and appears to activate pro MMP-2 which degrades collagen.

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Growth and Survival Rates of Flat Oyster, Ostrea denselamellosa, by Condition of Larval Cultivation (벗굴 (Ostrea denselamellosa) 유생의 생육조건에 따른 성장과 생존)

  • Yang, Moon-Ho;Oh, Bong-Se;Han, Chang-Hee
    • The Korean Journal of Malacology
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    • v.19 no.2
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    • pp.133-142
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    • 2003
  • For the effective seedling production of flat oyster, Ostrea denselamellosa, dietary value of live food, densities, water temperature and salinity on growth and survival rate of the larvae were examined. In rearing larvae by feeding them phytoplankton diets, the optimal survival rate and growth rate of larvae were found using a mixed phytoplankton diet which was mixed with Isochrysis galbana, Chaetoceros calcitrans and Chlorella sp. The highest growth and survival rates of the larvae were 208.4% and 38.8% with the phytoplankton diet. In growth and survival rates of larvae with various rearing densities, the highest survival and growth rates were 228.1% and 29.0% at the density of 2 individuals/ml. In observing rearing experiments of the flat oyster larvae under various temperature conditions, average growth rates of the larvae in respect to shell length were 202.2%, 240.4%, 250.6% and 121.3% in natural water temperatures (18-22$^{\circ}C$), 24$^{\circ}C$, 28$^{\circ}C$ and 32 $^{\circ}C$, respectively. And average survival rates of the larvae were 16.0%, 32.0%, 13.0% and 0% in natural water temperatures (18-22$^{\circ}C$), 24$^{\circ}C$, 28$^{\circ}C$ and 32$^{\circ}C$, respectively. In rearing at various salinities, the highest growth rates of the larvae in shell length was 240.0% at 30.0 psu and the highest survival rate was 31.0% at 25 psu.

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Effects of Gonadotropin Releasing Hormone on Steroidogenesis and Apoptosis of Human Granulosa-Lutein Cells (생식샘자극호르몬분비호르몬이 사람 과립-황체화 세포의 스테로이드 생성과 세포자연사에 미치는 영향)

  • Lee, Hyo-Jin;Yang, Hyun-Won
    • Development and Reproduction
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    • v.13 no.4
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    • pp.353-362
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    • 2009
  • GnRH and its receptor are known to express locally in the ovary and to regulate the ovarian function by affecting on granulosa and lutein cells. It has been reported that GnRH directly causes apoptosis in the granulosa and lutein cells of the ovary. However, whether the apoptosis of the cells by GnRH is recovered by FSH as an anti-apoptotic factor is not yet known. In this study, we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ after treatment with 5, 50, and 100 ng/$m\ell$ GnRH and 1 IU/ml FSH in the granulosa-lutein cells that are obtained during oocyte-retrieval for IVF-ET. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in the cells treated with 100 ng/$m\ell$ GnRH. In addition, we found that FSH suppresses the apoptosis of the cells induced by GnRH. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by GnRH treatment, whereas $E_2$ production was not changed. We also demonstrated that FSH inhibits the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present results suggest that GnRH agonist using in ovarian hyperstimulation protocol might induce the dysfunction of the ovary, but its function could be recovered by FSH. These results also will be expected to use as the basic data to elucidate the physiological role of GnRH and to develop new ovarian hyperstimulation protocols for IVF-ET.

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Role of Ghrelin in the Control of Reproductive Endocrine Function (포유류 생식 내분비 기능 조절에서 Ghrelin의 역할)

  • Lee, Sung-Ho
    • Development and Reproduction
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    • v.13 no.4
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    • pp.207-215
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    • 2009
  • Numerous factors can affect the activities of hypothalamus-pituitary-gonad (HPG) hormonal axis, resulting in alteration of reproductive capacity or status such as onset of puberty and menopause. Soon after the finding of leptin, a multifunctional hormone secreted from adipocytes, a close relationship between reproduction and body energy balance have been manifested. Ghrelin, another multifunctional hormone from gastrointestinal tract, is an endogenous ligand of growth hormone secretagogue receptor (GHSR), and is thought to be a counterpart of leptin in the regulation of energy homeostasis. As expected, ghrelin can also modulate the reproductive capacity through the modulation of activities of HPG axis. This paper summarizes the current knowledge on the discovery, gene structures, tissue distribution and roles of ghrelin and GHSRs in mammalian reproduction in particular modulation of reproductive hormone secretion in HPG axis. Like POMC gene expression in pituitary gland, preproghrelin gene can generate a complex repertoire of transcripts which further undergo alternative splicing and posttranslational modifications. Concerning the roles of preproghrelin gene products in the control of body physiology except energy homeostasis, limited knowledge is available so far. Several lines of evidence, however, show the interplay of ghrelin between metabolism and reproduction. In rat and human, the distribution of ghrelin receptor GHSRs (GHSR1a and GHSR1b) has been confirmed not only in the hypothalamus and pituitary which were originally postulated as target of ghrelin but also in the testis and ovary. Expression of the preproghrelin gene in the brain and gonads was also verified, suggesting the local role (s) of ghrelin in HPG axis. Ghrelin might play a negative modulator in the secretions of hypothalamic GnRH, pituitary gonadotropins and gonadal steroids though the action on pituitary is still questionable. Recent studies suggest the involvement of ghrelin in regulation of puberty onset and possibly of menopause entry. It is now evident that ghrelin is a crucial hormomal component in 'brain-gut' axis, and is a strong candidate links between metabolism and reproduction. Opposite to that for leptin, ghrelin signaling is likely representing the 'hunger' state of body energy balance and is necessary to avoid the energy investment into reproduction which has not a top priority in maintaining homeostasis. Further researches are needed to gain a deep insight into the more precise action mechanism and role of ghrelin in reproduction, and to guarantee the successful biomedical applications.

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Embryonic and Larva Development of Slime Flounder, Microstomus achne (찰가자미(Microstomus achne)의 난발생 및 자치어 형태 발달)

  • Byun, Soon-Gyu;Lee, Sung-Hun;Hwang, Jae-Ho;Han, Kyeong-Ho;Kang, Kyeong-Wan;Kim, Jin-Do;Kim, Yi-Cheong;Lee, Bae-Ik
    • Development and Reproduction
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    • v.13 no.4
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    • pp.281-289
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    • 2009
  • The slime flounder Microstomus achne were caught at Geomun Island, Yosu-si, Jeollanamdo from January to March in 2006. The fertilized eggs were observed for morphological development of egg, embryo and larva. Eggs were colorless transparent, separative pelagic, absent of oil globule, and the diameter was 1.64${\pm}$0.03 mm (n=50). The eggs were hatched at 168 hours 40 minutes after fertilization in the range of $9.8\sim13.0^{\circ}C$ (mean $11.4{\pm}1.6^{\circ}C$). Total length of newly hatched larva was 4.05${\pm}$0.18 mm (n=20). The larva had developed membranous fin showing waterdrop-shaped structure, and their mouth and anus were not open. The myotomes were 14~15+33~34=47~49. The egg yolks were 1.64${\pm}$0.12 mm in major axis, and 1.23${\pm}$0.19 mm in minor axis. At 12 days after hatching, the total length was 7.32${\pm}$0.42 mm(n=20). The egg yolk was completely absorbed and transferred to post larval stage. Star-shaped melanophores and branch-shaped xanthophores in the edge of membranous fin were more densed. Chrysanthemum-shaped melanophores in the notochord were densed and formed 4~5 melanophore bands. At 90~93 days after hatching, morphological features of the larva, 19.91${\pm}$1.63 mm TL(n=20), were transferred to juvenile stage showing similar features with those of the adult fish.

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Hatching Rate of Eggs, and Growth of Larvae and Juveniles from Selected Olive Flounder, Paralichthys olivaceus (선발 육종된 넙치, Paralichthys olivaceus의 부화율 및 자치어 성장)

  • Min, Byung-Hwa;Lee, Jeong-Ho;Noh, Jae-Koo;Kim, Hyun-Chul;Park, Choul-Ji;Choi, Sang-Jun;Myeong, Jeong-In
    • Development and Reproduction
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    • v.13 no.4
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    • pp.239-247
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    • 2009
  • Hatching rate, larval deformation and growth rate of selected olive flounder (Paralichthys olivaceus) for rapid growth were compared to those of the unselected fish. Fish were spawned on the same day and cultured under the similar conditions. The selected fish had a significantly higher eggs hatching rate, and lower larval deformation. The selected fish grew significantly faster, and at the end of the experiment (8 weeks after hatching) averaged 50.49${\pm}$2.67 mm in total length, 16.30${\pm}$0.08 mm in body height, and 1.036${\pm}$0.118 g in weight, compared to 40.55${\pm}$3.13 mm, 13.50${\pm}$0.96 mm, and 0.557${\pm}$0.073 g for unselected fish, respectively. The selected fish had a significantly higher body shape index, however lower condition factor than the unselected fish. The results of the present study demonstrate that the selected fish of the olive flounder for rapid growth had superior growth and improved body shape.

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Effects of Cortisol on the Steroidogenesis and the Apoptosis of Human Granulosa-Lutein Cells (Cortisol이 사람 과립-황체화 세포의 스테로이드 생성과 세포자연사에 미치는 영향)

  • Kim, Jin-Hee;Yang, Hyun-Won
    • Development and Reproduction
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    • v.13 no.4
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    • pp.329-339
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    • 2009
  • Cortisol is present in high concentration in the ovary and its receptor is expressed in the ovarian cells. Moreover, cortisol is known to have a role in steroid synthesis and cell metabolism in human granulosa and lutein cells. However, little is known of the role of cortisol presenting in high concentration in the follicles after LH surge on the granulosa-lutein cells. Therefore, the this study we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ in the granulosa-lutein cells that are obtained during oocyte-retrieval after treatment with 5, 50, and $500{\mu}g/m\ell$ cortisol and 1 IU/$m\ell$ FSH. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in 50 and $500{\mu}g/m\ell$ cortisol treated cells. We found, however, that FSH did not suppress the apoptosis of the cells induced by cortisol. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by cortisol treatment, whereas $E_2$ was not changed. We also demonstrated that FSH did not inhibit the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present study suggests that cortisol of high concentration could cause the apoptosis of human granulosa-lutein cells by suppressing the production of $P_4$. However, we need more studies to elucidate the mechanism by which cortisol induces apoptosis in human granulosa-lutein cells in view of the fact that our results are inconsistent with previous reported data.

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Effect of Stem Cell-Derived Conditioned Medium on the In Vitro Maturation and Embryonic Development of Parthenogenetic Embryos in Pigs (Stem Cell-Derived Conditioned Medium 첨가가 돼지난자의 체외성숙 및 단위발생란의 초기배 발육에 미치는 영향)

  • Kwon, Dae-Jin;Hwang, In-Sul;Kwak, Tae-Uk;Oh, Keon Bong;Ock, Sun-A;Chung, Hak-Jae;Im, Gi-Sun;Hwang, Seongsoo
    • Reproductive and Developmental Biology
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    • v.39 no.3
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    • pp.89-95
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    • 2015
  • The addition of growth factors and cytokines to in vitro culture (IVC) media could affect embryo development and the quality of the resulting blastocysts. The present study was performed to investigate the effect of porcine induced pluripotent stem cell (piPSC)-culture conditioned medium (CM) on the in vitro maturation (IVM) and development of parthenogentic embryos (parthenotes) in pigs. Cumulus-oocyte complexes (COCs) or activated oocytes were cultured in IVM or IVC medium supplemented with 0 (control), 25, or 50% of stem cell medium (SM) or CM, respectively. The maturation rate of CM-25% group was significantly improved when compared with control group (p<0.05), but that was not different among SM or CM groups. Blastocyst formation rate was significantly higher in CM-25% group (29.2%) than that of control (20.7%), SM-50% (19.6%) and CM-50% (23.66%, p<0.05). Cell number and the apoptotic cell index in blastocysts was significantly lower in SM-25% than in CM-25% group (p<0.05). The embryo quality related genes, OCT4, KLF4, TERT and ZFP42, were significantly increased in CM-25% group compared with control (p<0.05). In conclusion, the addition of 25% of CM to IVM and IVC medium positively influences not only the developmental potential also quality of parthenotes in pig.

Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Effects of Gestation Length and Birth Weight on Survival Rate in Cloned Korean Native Calves (복제 송아지의 임신 기간과 생시체중이 출생 후 생존율에 미치는 영향)

  • Yang, Byoung-Chul;Im, Gi-Sun;Kim, Dong-Hoon;Ko, Yeoung-Gyu;Hwang, Seong-Soo;Nho, Whan-Gook;Kim, Myung-Jick;Yang, Boh-Suck;Lee, Sang-Jin;Seong, Hwan-Hoo
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.51-58
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    • 2008
  • This study was performed to investigate the relation between birth weight and survivability on the production of cloned Hanwoo calves. The 580 cloned embryos were transferred into the 293 recipients. The pregnancy rate of the cloned embryos was 72.3% at 50 days after embryo transfer, and then the rate was dramatically decreased. The mean gestation lengths were 287 days in both clone (range of$279{\sim}295$ days) and artificial insemination (AI, range of $255{\sim}293$ days) calves, respectively. The mean birth weight of cloned calves (30.3kg) was significantly higher compared to that of AI calves (23.7kg) (p<0.05). Among the cloned calves, the birth weight was not different in both normal delivery (n=17, 29.9kg) and caesarean section (n=14, 32.3kg). The weight, however, was significantly higher in the clones (n=18, 32.8kg) dead within 175 days than that of the clones (n=11, 28.3kg) alive more than 175 days after birth (p<0.05). Interestingly, all cloned calves weighed <15kg (n=5) or >35kg (n=9) at birth have been dead within 175 days from the date of birth. The causes of death in the cloned calves were premature birth (n=2, 10.0%), abnormal function of lung and liver (n=2, 10.0%), abnormal function of lung (n=4, 20.0%), malformation (n=4, 20.0%), unknown (n=4, 20.0%), and sudden death syndrome (n=4, 20.0%), respectively. Our findings suggest that normal birth weight is one of the most important factors to survive more than 6 months in cloned calves.