The present study investigated the effects of follicle stimulating hormone (FSH) and human chorionic gona-dotrophin (hCG) on the nuclear maturation of canine oocytes. Oocytes were recovered from mongrel female ovaries in various reproductive states; follicular, luteal or anestrous stage. Oocytes were cultured in sserum-free tissue culture medium (TCM)-199 supplemented with various concentrations of FSH (Exp. 1: 0, 0.5, 1.0 or 10 IU) or hCG (Exp.2:0, 0.5, 1.0 or 10 IU) or both (Exp. 3:1 IU FSH +1 IU hCG) for 72 hr to determine the effective concentration of these hormones, and to examine their combined effect. After maturation culture, oocytes were denuded in PBS containing 0.1% (w/v) hyaluronidase by gentle pipetting. The denuded oocytes were stained with $1.9\;{\mu}M$. Hoechst 33342 in glycerol and the nuclear state of oocytes was evaluated under UV light. More (p<0.05) oocytes matured to MII stage when follicular stage oocytes were supplemented with 1 IU FSH (6.2%) compared with the control, 0.1 or 10.0 IU FSH (0 to 1.2%). Significantly higher (p<0.05) maturation rate to MII stage was observed in follicular stage oocytes supplemented with 1.0 IU hCG (7.2%) compared with the control or other hCG supplemented groups (0 to 1.5%). However, the combination of FSH and hCG did not improve the nuclear maturation rate of canine oocyte (2.4 %) compared with FSH (6.2%) and hCG alone (7.2%). In conclusion, FSH or hCG alone significantly increased the maturation of canine oocytes to MII stage.
The polysialic acid (polySia) glycotope covalently modifies cell surface glycoconjugates on cells as evolutionarily diverse as microbes and human. The recent chemical identification of polysialylated glycoproteins in the jelly coat and on the cell surface of the sea urchin egg raises important questions about their biosynthesis and possible function. Using CMP-[$^{14}$ C]Neu5Ac as substrate and cell free preparations from eggs and embryos of the sea urchin Stronglylcentrotus nudus, we have identified a membrane associated CMP-Neu5Ac:poly-$\alpha$2, 8 sialosyl sialyltransferase (polyST) that transfers Neu5Ac to an endogenous acceptor. Optimal conditions for the polyST activity were found to be 2$0^{\circ}C$ in 20 mM MOPS buffer (pH 7.0). The polyST activity was increased 2.7 times by the addition of 10 mM $Mg^2$$^{+}$. The membrane-associated polyST also catalyzed the polysialylation of mammalian ganglioside GD3. Given that no structurally similar natural polysialylated gangliosides have been described, nor were observed in the present study, we conclude that a single polyST activity catalyzes sialylation of the endogenous acceptor and the gangliosides. Using an excess of GD3 as an exogenous acceptor, it was established that the expression of the polyST in S. nudus embryos increased rapidly at the mesenchyme blastula stage and reached at maximum at the gastrula stage. The finding that this polyST in the sea urchin embryo is developmentally regulated raises the possibility that it may play a role in the changing cell and tissue interactions that occur during gastrulation and the early stages of spicule formation.n.
The effect of nitric oxide (NO) on antioxidant system and protective mechanism against oxidative stress under UV-B radiation was investigated in leaves of maize (Zea mays L.) seedlings during 3 days growth period. UV-B irradiation caused a decrease of leaf biomass including leaf length, width and weight during growth. Application of NO donor, sodium nitroprusside (SNP), significantly alleviated UV-B stress induced growth suppression. NO donor permitted the survival of more green leaf tissue preventing chlorophyll content reduction and of higher quantum yield for photosystem II than in non-treated controls under UV-B stress, suggesting that NO has protective effect on chloroplast membrane in maize leaves. Flavonoids and anthocyanin, UV-B absorbing compounds, were significantly accumulated in the maize leaves upon UV-B exposure. Moreover, the increase of these compounds was intensified in the NO treated seedlings. UV-B treatment resulted in lipid peroxidation and induced accumulation of hydrogen peroxide ($H_2O_2$) in maize leaves, while NO donor prevented UV-B induced increase in the contents of malondialdehyde (MDA) and $H_2O_2$. These results demonstrate that NO serves as antioxidant agent able to scavenge $H_2O_2$ to protect plant cells from oxidative damage. The activities of two antioxidant enzymes that scavenge reactive oxygen species, catalase (CAT) and ascorbate peroxidase (APX) in maize leaves in the presence of NO donor under UV-B stress were higher than those under UV-B stress alone. Application of 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3- oxide (PTIO), a specific NO scavenger, to the maize leaves arrested NO donor mediated protective effect on leaf growth, photosynthetic pigment and free radical scavenging activity. However, PTIO had little effect on maize leaves under UV-B stress compared with that of UV-B stress alone. $N^{\omega}$-nitro-L-arginine (LNNA), an inhibitor of nitric oxide synthase (NOS), significantly increased $H_2O_2$ and MDA accumulation and decreased antioxidant enzyme activities in maize leaves under UV-B stress. This demonstrates that NOS inhibitor LNNA has opposite effects on oxidative resistance. From these results it is suggested that NO might act as a signal in activating active oxygen scavenging system that protects plants from oxidative stress induced by UV-B radiation and thus confer UV-B tolerance.
Journal of the korean academy of Pediatric Dentistry
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v.24
no.1
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pp.204-219
/
1997
The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.
Journal of the Korean Society of Food Science and Nutrition
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v.43
no.4
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pp.612-617
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2014
Anti-inflammatory effects of turmeric (Curcuma longa L.) rich in polysaccharides, as well as free of curcuminoids and turmerones were investigated in acute and chronic inflammatory models. Activity against the acute phase of inflammation was evaluated in carrageenan-induced paw edema and xylene-induced ear edema models. The results showed that turmeric extract significantly decreased paw edema volume in the first and third hours after carrageenan injection ($P{\leq}0.05$). Turmeric extract at all dose levels also significantly inhibited xylene-induced ear edema formation ($P{\leq}0.05$). Activity against chronic inflammation was also evaluated in cotton pellet-induced granuloma model. Turmeric extract significantly ($P{\leq}0.05$) decreased the weight of granuloma tissue on cotton pellets in a dose-dependent manner when compared to the vehicle control. Thus, the findings of the study suggest that turmeric extract in effective against both acute and chronic inflammation.
Five kinds of sensory cells, called A1-, A2-, B-, C-, and D-type cell, respectively, are observed in the epithelial tissue of suker's infundibulum of Cephalopoda, Octopus minor. The A1-type cells lie side by with the B-type cell in the epithelium of sucker's infundibulum. In the A1-type, the nucleus shapes irregularly and the karyolymph appears dark due to its high electron density. The cytoplasm is filled with many vacuoles of various sizes ($0.04\sim0.4{\mu}m$ in diameter), which move to the apical portion of the cell to be secreted via glycocalyx. The A2-type cells are mainly found at the basal portion of the epithelium. The shape of its nucleus is similar to that in the A1-type cell, and the cytoplasm, filiform or in reticular form, shows high electron density. The B-type cell contains an ovoid nucleus and the cytoplasm where lots of vacuoles which resemble the endoplasmic reticulum and electron-dense round granules of various sizes $(0.25\sim0.6{\mu}m)$ are found. The vacuoles and granules are secreted into the free surface via glycocalyx. The C- and D-type cells in simple or stratified layer are observed at the folded portion of the sucker's epithelium. The C-type cell contains a low electron-dense elliptical nucleus, while the D-type cell has an irregular nucleus where beterochromatin is well developed.
Objectives : We studied Effects of Hagocho(prunella vulgaris L.), Gamgook(chrysanthemum indicum L.) and Galgeun(pueraria Radix) aqua-acupuncture on the hyperlipidemic rat. methods : We investigated lowering lipid effect, oxidative capacity, concentration of TNF-${\alpha}$, IL-6, Leptin and histological consideration in hyperlipidemic rat. Forty male Sprague-Dawley rats weighing about 400g were divided into 5 groups of control, Ⅰ: Hagocho (prunella vulgaris L.)+Gamgook (chrysanthemum indicum L.) and Gokji aqua-acupuncture, Ⅱ: Hagocho(prunella vulgaris L.)+Gamgook (chrysanthemum indicum L.) and Joksamri aqua-acupuncture, Ⅲ: Hagocho(prunella vulgaris L.)+Galgeun(pueraria Radix)and Gokji aqua-acupuncture and Ⅳ: Hagocho (prunella vulgaris L.)+Galgeun(pueraria Radix) and Joksamri aqua-acupuncture. Results : Contents of plasma ${\beta}$-lipoprotein, contents of IL-6, TNF-${\alpha}$ and leptin, Plasma triglyceride and glucose, plasma total cholesterol and LDL-cholesterol, liver total cholesterol, liver triglyceride, plasma and liver TBARS, free fatty acids showed a tendency to decrease in the aqua-acupuncture groups compared to those of control group. The activities of GOT and GPT showed no significantly different in all treatment groups. Values of super oxide dismutase, glutathione peroxidase and catalase activity showed a tendency to increase in the aqua-acupuncture groups. Histological consideration of heart, kidney and liver in the aqua-acupuncture groups showed slight vasodilation and fat accumulation compared to those of normal rat. Conclusions : These results indicated that prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix aqua-acupuncture at gokji(LI11) and Joksamri(ST36) suppressed adipose tissue mass and lipid peroxidation and increased antioxidant system in hyperlipidemic rat.
Objectives : This study was carried out to identified the effects of distilled cultivated wild ginseng pharmacopuncture to the obesity. Methods : Cultivated wild ginseng pharmacopuncture was administered on the points of chung-wan(CV12), $Ch'{\breve{o}}nch'u$(ST25), and Chok-samni(ST36) on lowering lipid and oxidative capacity in biochemical and molecular biological aspects were investigated in obese rats fed with high fat diet. Results : 1. The contents of plasma ${\beta}-lipoprotein$ showed a tendency to decrease in the pharmacopuncture groups compared to the control group. In the pharmacopuncture groups, the values of ST25 and ST36 pharmacopuncture groups showed lower value. 2. The contents of plasma free fatty acids showed a tendency to decrease in pharmacopuncture groups compared to the control group. However, in the pharmacopuncture groups, the values were not significantly different. 3. Plasma triglyceride and glucose showed lower value in the ST25 pharmacopuncture groups compared with the other groups. 4. The activity of AST showed a tendency to decrease in the pharmacopuncture groups. However, the activity of ALT was not significantly different in all the treatment groups. 5. Plasma total cholesterol and LDL-cholesterol showed lower value in the ST25 and ST36 pharmacopuncture groups and HDL-cholesterol showed higher value in the CV12 pharmacopuncture groups than that of the other treatment groups. 6. Liver total cholesterol values didn't show significant difference in all the treatment groups, and triglyceride showed lower value in the pharmacopuncture groups. 7. The contents of plasma TEARS showed lower value in the ST25 pharmacopuncture group and contents of liver TBARS showed a tendency to decrease in the pharmacopuncture groups. However these values didn't show significant difference in the pharmaco puncture groups. 8. Liver super oxide dismutase activity showed higher value in the ST25 and ST36 pharmacopuncture groups, and the value of liver glutathione peroxidase and catalase activity showed a tendency to increase in the pharmacopuncture groups. However, these values showed no significant difference in the pharmacopuncture groups. 9. Expression of apo-B and E mRNA in liver cells was lower in the ST25 pharmacopuncture group than that of the other treatment groups. However, expression of $TNF-{\alpha}$ and leptin mRNA in adipose cell showed no difference among all the treatment groups. 10. ST25 pharmacopuncture group showed a good histological character of liver. It showed similar to that of normal group. However other treatment groups and control group showed slight vasodilation and slight fat accumulation. Conclusion : These results indicate that distilled cultivated wild ginseng pharmacopuncture suppressed adipose tissue mass and lipid peroxidation, and increased antioxidant capacity.
The lipid lowering and antioxidant effects of Gansoo(BL18), Pungji(GB20) and Eumnungcheun(SP9) acupuncture in rats fed high fat diet were analyzed in biochemical and molecular biological aspects. The results obtained from this study are as follows : 1. In the body weight reduction, all acupuncture groups showed a high reduction compared to those of control group and in acupuncture groups, Gansoo(BL18) and Pungji(GB20) acupuncture groups showed a high reduction. 2. The concentration of plasma triglyceride, total cholesterol and LDL-cholesterol with acupuncture groups showed a little decrease and in acupuncture groups, Gansoo(BL18) and Pungji(GB20) groups showed a low values compared to those of other acupuncture groups. However, the tendency of HDL-cholesterol concentration showed no significant different. 3. The concentration of plasma ${\beta}-lipoprotein$ and free fatty acids showed a lowest values in the Gansoo(BL18) and Pungji(GB20) acupuncture groups and the glucose concentration showed to decrease in all treated acupuncture groups. 4. The concentration of liver total cholesterol and triglyceride in Gansoo(BL18) and Pungji(GB20) acupuncture groups showed a lower values than those of control group. 5. In all the acupuncture groups, the plasma glutamic oxaloacetic transaminase(GOT) activity showed a little decrease. In the glutamic pyruvate activity(GPT), Gansoo(BL18) and Pungji(GB20) acupuncture groups showed a lower values than those of control groups. However the values of eumneungcheun acupuncture only group showed no significant difference to those of control group. 6. The plasma and liver Thiobabituric acid reactive substance (TBARS) concentration in Gansoo(BL18) and Pungji(GB20) acupuncture groups were a lower than those of control group. However the values of eumneungcheun acupuncture group showed no significant difference to control group. 7. The superoxide dismutase (SOD) activity in Gansoo(BL18) acupuncture group and Glutathione peroxidase (GSH-Px) activity in Gansoo(BL18) and Pungji(GB20) acupuncture groups showed a high values. The catalase (CAT) activity in all the acupuncture groups showed a higher values than those of control group. 8. In acupuncture groups, DNA expression of Apo-B and Apo-E showed a tendency to decrease, however DNA expression of leptin showed no significant difference in all treatment groups. DNA expression of $TNF-{\alpha}$ showed a increase in acupuncture groups. These results indicate that Gansoo(BL18) and Pungji(GB20) (especially Gansoo(BL18)) acupuncture affect the lipid metabolism and showed possibility of lowering adipose tissue mass and lipid peroxidation.
Journal of Physiology & Pathology in Korean Medicine
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v.27
no.5
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pp.611-616
/
2013
Alcoholic liver disease (ALD) is a major cause of morbidity and mortality around the world. Although much progress has been made in understanding the pathogenesis of ALD, there remains no effective therapy for it. Accumulated evidence indicates that oxidative stress is the main pathological factors in the development of ALD. Ethanol administration causes accumulation of reactive oxygen species (ROS), including superoxide, hydroxyl radical, and hydrogen peroxide. ROS, in turn, cause lipid peroxidation of cellular membranes, and protein and DNA oxidation, which results in hepatocyte injury. In addition to pro-oxidants formation, antioxidants depletion caused by ethanol administration also results in oxidative stress. The objective of this study is to investigate the effects of Shiryung-tang extract on the chronic alcoholic liver injury induced by EtOH. Male Sprague Dawley rats were used in this study. All rats were maintained under standard laboratory conditions ($23{\pm}1^{\circ}C$, 12h light/12h dark cycles). All animals (n=30) were randomly divided into following groups: (1) Normal group, treated with distilled water (n=10); (2) Control group, treated with ethanol (n=10); (3) Sample group, treated with ethanol + pharmacopuncture (n=10). For oral administration of ethanol in Control and Sample group, the ethanol was dissolved in distilled water in concentrations of 25%(v/v). Throughout the experiment of 8 week, the rats were allowed free access to water and standard chow. Sample group were administrated by Shiryung-tang extract daily for 8 weeks. Control group were given normal saline for same weeks. As a results, the oral administration of ethanol for 8 weeks leads to hepatotoxicity. The levels of hepatic marker such as HDL-cholesterol, triglyceride, aspartate aminotransferase and alanine aminotransferase were altered. The ethanol also increased lipid peroxidation and depletion of antioxidant enzyme activities as well as hepatic tissue injury. However, the treatment of Shiryung-tang extract prevented all the alterations induced by ethanol and returned their levels to near normal. These data suggest that Shiryung-tang extract could have a beneficial effect in inhibiting the oxidative damage induced by chronic ethanol administration. Therefore, Shiryung-tang extract can be a candidate to protect against EtOH-induced liver injury.
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