In this study, we investigated the effects of Undaria pinnatifida (UP), Petalonia binghamiae (PB) and Punctaria latifolia (PL) extracts on the inhibition of proliferation and apoptosis in human gastric and breast cancer cells. AGS, MDA-MB-231 and SK-BR-3 cells were treated with 0, 50, 100, and 200 μg/ml concentrations of the extracts to determine their anti-proliferative effects, using the MTT assay. The UP, PB and PL extracts inhibited proliferation of AGS, MDA-MB-231 and SK-BR-3 cells in a dose-dependent manner, and the PL extract was found to be the most effective. DAPI staining was also performed to determine changes in the cell nucleus. Further, the AGS, MDA-MB-231 and SK-BR-3 cells were treated with 0, 50, 100, and 200 μg/ml of only the PL extract. DAPI staining showed increased chromatin condensation, which is indicative of apoptosis, in the 200 μg/ml group. The expression of the Bax, Bcl-2, and PARP proteins in AGS, MDA-MB-231 and SK-BR-3 cells treated with the PL extract was also determined by western blot analysis. The expression of Bax (a pro-apoptotic protein) and cleaved-PARP was increased, whereas the expression of Bcl-2 (an anti-apoptotic protein) was decreased compared with the control. These findings indicate that the PL extract may have potential as an alternative anticancer drug and nutraceutical.
This experiment was carried out to evaluate the effect of mouse early embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells(BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined. For a comparative study of in vi패 and in vitro development, the fresh blastocyst which developed in vivo for 120 hours after hCG injection was collected from the uterus, and their numbers of nuclei were also counted. The higher developmental rates of blastocyst formation was a, pp.ared from 91% to 97% when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal eptithelial cells. The number of nuclei in the embryos cultured for 72 hours under each conditions was significantly reduced it than blastocyst in vitro conditions. The number of nuclei in embryos cultured in TCM 199, Ham's F-10 and Medicult IVF medium were counted 68.1$\pm$6.00, 67.3$\pm$4.49, 66.4$\pm$5.64, and 94.3$\pm$8.61, 92.5$\pm$7.60, 92.1$\pm$6.10 with BOEC and 93.3$\pm$5.80, 92.9$\pm$6.53, 92.3$\pm$7.35 with POEC coculture, respectively. These numbers were lowered than 107.2$\pm$7.43 in vivo conditions. In conclusions, the coculture between the mouse early embryos, and oviductal epithelial cells of BOEC and POEC give to improve the developmental and hatching rates of blastocyst but in vivo culture systems for the growth of nuclei were ineligible than in vitro conditions.
Purpose: We investigated whether a relationship exists between tumor control dose 50 ($TCD_{50}$) or tumor growth delay (TGD) and radiation induced apoptosis (RIA) in syngeneic murine tumors. Also we investigated the biological markers that can predict radiosensitivity in murine tumor system through analysis of relationship between $TCD_{50}$, TGD, RIA and constitutive expression levels of the genetic products regulating RIA. Materials and Methods: Syngeneic murine tumors such as ovarian adenocarcinoma, mammary carcinoma, squamous cell carcinoma, fibrosarcoma, hepatocarcinoma were used In this study. C3H/HeJ mice were bred and maintained in our specific pathogen free mouse colony and were $8{\sim}12$ weeks old when used for the experiments. The tumors, growing in the right hind legs of mice, were analyzed for $TCD_{50}$, TGD, and RIA at 8 mm in diameter. The tumors were also analyzed for the constitutive expression levels of $p53,\;p21^{WAF1/CIP1},\;BAX,\;Bcl-2,\;Bcl-X_L,\;Bcl-X_S$, and p34. Correlation analysis was peformed whether the level of RIA were correlated with $TCD_{50}$ or TGD, and the constitutive expression levels of genetic products regulating RIA were correlated with $TCD_{50}$, TGD, RIA. Results: The level of RIA showed a significant positive correlation (R=0.922, p=0.026) with TGD, and showed a trend to correlation (R=-0.848), marginally significant correlation with $TCD_{50}$ (p=0.070). It indicates that tumors that respond to radiation with high percentage of apoptosis were more radiosensitive. The constitutive expression levels of $p21^{WAF1/CIP1}$ and 34 showed a significant correlation either with $TCD_{50}$ (R=0.893, p=0.041 and R=0.904, p=0.035) or with TGD (R=-0.922, p=0.026 and R=-0.890 p=0.043). The tumors with high constitutive expression levels of $p21^{WAF1/CIP1}$ or p34 were less radiosensitive than those with low expression. Conclusion: Radiosensitivity may be predicted with the level of RIA in murine tumors. The constitutive expression levels of $p21^{WAF1/CIP1}$ or p34 can be used as biological markers which predict the radiosensitivity.
To investigate the toxicological effects of ${\beta}$-glucan, we performed basic studying on ${\beta}$-glucan in Sprague-Dawley (SD) rats. Standard endpoints in this study included mortality, clinical observations, changes of body weights, analysis on food consumption, ophthalmoscopic examination, hematologic examination, serum biochemistry, analysis of organ weights, gross anatomic pathology and histopathology. No clinical signs and mortality were observed in animals treated with beta-glucan throughout the experimental period. The average body weight of each treatment groups showed similar levels at end of experiment. There were no treatment-related changes in mortality, body weights changes, food consumption, ophthalmoscopic examination. Although there were statistically significant differences between the control and treated groups in some relative and absolute organ weights, and hematological and biochemical analysis, the data were in biologically normal ranges and did not show a dose-dependent manner. In the morphological change, hepatic tissue were not showed ballooning degeneration and irregular arrangement of hepatic cell in ${\beta}$-glucan treatment groups with control group. Also, organs weights (liver, heart, kidney and stomach) and hematological indices (WBC, RBC, Hb, Hct and Platelet) did not show statistically significant differences among the experimental groups. In summary of these results, there were no changes in mortality, mean body weight, clinical signs, food consumption. There were no changes in ophthalmological examination, hematology, blood chemistry, necropsy and histopathology. In conclusion, although further investigation of glucan should be performed in the functions registered in many ancient literatures, ${\beta}$-glucan is physiologically safe and may have potential as candidate food for human health.
Tomatoes are flooded differently 0, 5, 10 and 15 ㎝, according to the developing stages such as transplanting stage under the condition of green house. Along with this, they are treated according to the time condition such as 6, 12, 24, 48 and 120 hours. The results obtained are summarized as follows. As the depth of flooding got deeper and the hours got longer, plant height, number of leaves, shoot and root decreased significantly. Flowering was possible for 24 hours in the flooding of 0 ㎝, for 6 hours in $5{\sim}10$ ㎝, but not possible after 6 hours in 15 ㎝. Without regard to the depth of flooding, adventitious root came into being before or after 48 hours of the treatment. Root activity diminished gradually as hours of treatment went by, but diminished rapidly over the depth of 5 ㎝. Chlorophyll content decreased similarly as in the case of root activity. Diffusion resistance of stomata cell increased as hours of treatment passed and depth increased. Photosynthesis and respiration diminished according as the hours and depth of treatment increased. Respiration diminished a little gradually but photosynthesis weakened greatly as the depth of treatment became greater and after 48 hours of treatment. Diseases occurred remarkably in proportion to the depth of treatment and the increase of hours. The possibility of preventing by means of insecticide treatment showed the same tendency as in the seedling stage. But its effect was not significant. After 120 hours yields could not be expected because tomatoes died without regard to the depth of flooding. Instead of the depth, numbers of fruits per plant decrease of individuals or variation of average weight of a fruit was recognized. Especially average weight increased in accordance with the increase of the depth. There was positive correlation between all the characters, such as plant height, number of leaves, fresh weight, chlorophyll content, root activity and yield traits, but negative correlation between these and epinastic curvature, diffusion resistance and adventitious root.
Tomatoes are flooded differently 0, 5, 10 and 15cm, according to the developing stages such as flowering stage under the condition of greenhouse. Along with this, they are treated according to the time condition such as 6, 12, 24, 48 and 120 hours. The results obtained are summarized as follows. Plant height decreased in the depth of $0{\sim}10cm$ for over 48 hours, in the depth of 15cm for over 24 hours. Number of leaves was the same as in control, and it decreased over. Number of flowers and fruit setting of individuals decreased conspicuously according as the depth and the hours got greater and longer. Adventitious root occurred remarkably in the depth of $0{\sim}10cm$, for over 24 hours and in the depth of 15cm, 12 hours. Epinastic curvature increased greatly as the depth and the hours got greater and longer. Diffusion resistance of stomata cell increased as the depth and the hours got greater and longer. Diseases occurred conspicuously as the hours of flooding got longer rather than as the depth greater. The preventing of diseases caused by insecticide was observed, but it was not greater than in the seedling and transplanting stage. Fertilization was effective in the case of increasing the weight of shoot. Number of fruits per plant did not decrease in the depth of 0cm up to 24 hours, but decreased on the deeper level of flooding and increased as the hours got longer. Moreover with the exception of 120 hours per respective depth of the treatment, average weight of a fruit got greater as the depth and the hours got greater and longer. In the case of epinastic curvature and diffusion resistance, there was negative correlation between all the other investigated characters and positive correlation between weight of a fruits and average weight of a fruit.
In order to elucidate characteristics of water quality, investigation of monthly dynamics of environmental factors and algal populations at major four stations of the mid and lower part in Taechong Reservoir was performed from June 1998 to June 1999. Water temperature, DO and pH were ranged 5.3~27.7$^{\circ}C$, 6.2~13.8 mgO$_2$/1 and 6.4~9.5, respectively. Those were varying as the season changes. Among inorganic nitrogen nutrients, NH$_4$was ranged from 5.5% to 7.2% of NO$_3$and NO$_3$was almost same through the seasons except summer in which it was low. SRP and SRSi were increased in summer when the blue-green algae became dominant. Those were decreased as the cell density of diatom increased when the water temperature dropped. Therefore SRSi was considered to be another important nutrient factor contributing to the increment of biomass of freshwater algae as well as SRP. Average chi-$\alpha$ concentration ranged from s to 12 $\mu$g/1 and in the lower part or the reservoir, the lowest was found. Moreover, there were remarkable increment in summer when TN/TP ratio were decreasing from relationships between TN/TP ratio and chi-$\alpha$ concentration. Annual mean ratio of TN/TP ratio was relatively high as the value was 110, which was relatively high, and it showed that P is the dominant factor in the algal growth. The dynamics of phytoplankton were simply dominated by a few species seasonally. In summer, blue -green algae such as Anabaena, Microcystis and Oscillatoria were dominant and algal bloom of blue -green began from early summer, sustained to late autumn. The average standing crops of A. spiroides v. crassa, M. aeruginosa and O. limosa were ranged 0.3~2.0$\times$10$^4$cells/ml, 6.4$\times$10$^2$~1.0$\times$10$^4$cells/ml and 4.6$\times$10$^3$~1.6$\times$10$^4$cells/ml, respectively. In winter, diatom Stephanodiscus was considered to be an important species whose average standing crops of Stephunodiscus was 4.9$\times$10$^2$cells/ml from November to April of the next year and the highest was 1.3$\times$10$^3$cells/ml in January.
Purpose : To investigate the effects of angiotensin II inhibition on the epithelial to mesenchymal transition (EMT) in the developing kidney, we tested the expression of EMT markers and nestin in angiotensin converting enzyme (ACE) inhibitor-treated kidneys. Methods : Newborn rat pups were treated with enalapril (30 mg/kg/d) or a vehicle for 7 days. Immunohistochemistry for the expression of ${\alpha}$-smooth muscle actin (SMA), E-cadherin, vimentin, and nestin were performed. The number of positively-stained cells was determined under 100 magnification in 10 random fields. Results : In the enalapril-treated group, ${\alpha}SMA-positive$ cells were strongly expressed in the dilated tubular epithelial cells. The number of ${\alpha}SMA-positive$ cells in the enalapril-treated group increased in both the renal cortex and medulla, compared to the control group (P<0.05). The expression of E-cadherin-positive cells was dramatically reduced in the cortical and medullary tubular epithelial cells in the enalapril-treated group (P<0.05). The number of vimentin- and nestin-positive cells in the cortex was not different in comparisons between the two groups; however, their expression increased in the medullary tubulointerstitial cells in the enalapril-treated group (P<0.05). Conclusion : Our results show that ACE inhibition in the developing kidney increases the renal EMT by up-regulating ${\alpha}SMA$ and down-regulating E-cadherin. Enalapril treatment was associated with increased expression of vimentin and nestin in the renal medulla, suggesting that renal medullary changes during the EMT might be more prominent, and ACE inhibition might differentially modulate the expression of EMT markers in the developing rat kidney.
This study was conducted to investigate the hormonal changes in cultured medium during in vitro culture of bovine oviduct epithelial cells (BOEC) supplemented with interleukin (IL)-4 of 0.001, 0.01, 0.1 or 1 ng/ml. BOEC were collected from the oviduct and washed 3 times with 1% antibiotic-mycotic-DMEM medium and cultured at $39^{\circ}C$, 5% $CO_2$, 95% air for 24$\sim$120 hrs. The cultured media were analyzed hormonal changes with hormonal analyzing kit (progesterone (P4), estradiol (E2) : Perkin Elmer, USA) and Transforming growth factor (TGF)-$\beta$ with Eliza kit (Promega, USA). The production of P4 in 0.001 IL-4 was increased as the culture time increased. P4 production was significantly higher in the medium cultured for 120 hrs than 24 hrs (P<0.05). P4 production in 0.01 ng/ml group was similar to that of 0.001 ng/ml. The production of E2 in 0.001 and 0.01 ng/ml groups were increased to 72 hrs like P4 production and showed significantly different between the culture periods (P<0.05). After the culture for 96 hrs, P4 and E2 production were increased to 96 hrs, but decreased at 120 hrs. The production of TGF-$\beta$ showed no changes according to culture period or supplementation of IL-4. In conclusion, the supplementation of IL-4 can increase the production of P4 and E2 and might have important role for the successful pregnancy in bovine.
Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.
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