Indole 3-carbinol (I3C), important component of cruciferous vegetables and its major acid-catalyzed metabolite, 3,3'-diindolylmethane (DIM) have been suggested to have an inhibitory effect on the tumor growth and metastasis. This study investigated the effect of DIM on the adhesion, migration and invasion of highly invasive PC3 and DU145 human prostate cancer cell lines. Cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 3.0 g/L glucose, 3.7 g/L sodium bicarbonate and 10% fetal bovine and were incubated in a humidified incubator at $37^{\circ}C$ and 5% $CO_2$. DIM reduced the adhesion of PC3 and DU145 cells in a dose dependent manner. The pretreatment of PC3 cells with DIM reduced the adhesion dose dependantly, but inhibition was less effective than the treatment with DIM during the adhesion assay. The migration and invasion of PC3 and DU145 cells were reduced by DIM dose dependantly, and the inhibition of DIM was less effective in the DU145 cells than in the PC3 cells. The pretreatment of PC3 cells with DIM for 24 hr before the assay reduced invasion of PC3 cells by 37%. These results suggest that DIM inhibits adhesion, migration and invasion of the PC3 and DU145 cells and may be an effective antimetastatic therapy in addition to traditional chemotherapy.
Mineral trioxide aggregate (MTA) would influence healing of periapical tissues by modulating the production of growth factors and cytokines from PDL fibroblasts, however, the studies are insufficient. Therefore, the purpose of this study was to monitor the expression of transforming growth factor-beta1 $(TGF-\beta1)$, fibroblast growth factor-2 (FGF-2), and interleukin-6 (IL-6) from PDL fibroblasts in the presence of MTA. The human PDL fibroblasts were seeded onto the set MTA or IRM at a level of $1\times10^5$ cells per unit well, and further incubated for 6, 12, 24, and 48 hours. The levels of $TGF-\beta1$, FGF-2 and IL-6 from the supernatant were measured by enzyme-linked immunosorbent assay (ELISA) The data were analyzed using one-way ANOVA. The level of $TGF-\beta1$ was down-reg ulated when the cells were grown in the presence of MTA except at 6 hours. The levels of FGF-2 release were significantly suppressed when PDL fibroblasts were grown in the presence of MTA or IRM at all time intervals (p < 0.05). The expressions of IL-6 from MTA treated co)Is were comparable to those of untreated control cells throughout the observation periods. We presume that this material inhibits the stimulatory function of growth factors on granulation tissue formation and in turn, it promotes the healing process modulated by other bone-remodeling cells.
Experiments were conducted to determine the effects of beta-mercaptoethanol(${\beta}-ME$) supplements to the maturation medium on in vitro fertilization(IVF) and intracellular glutathione(GSH) concentration. Bovine cumulus-intact oocytes were matured in TCM-199 medium containing FBS, hormonal supplements, and ${\beta}-ME$(0, 25 and $50\;{\mu}M$) for 12h and 24 h. After culture, cumulus-free matured oocytes were co-incubated with frozen-thawed spermatozoa for 24h. Maturation rate increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant differences among treatment groups. Also, increases(p<0.05) in intracellular GSH concentration before and after fertilization were observed in $50\;{\mu}M\;{\beta}-ME$ supplements to the maturation medium. Male pronuclear formations after IVF was increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant difference among treatment groups. In conclusion, supplementing ${\beta}-ME$ into the maturation medium increased maturation rates, fertilization rates, and intracellular GSH concentrations.
Rice wine cake (RWC) is the solid waste obtained after rice wine fermentation. For the mass production of the spores of yeast Saccharomyces from RWC, the optimum pretreatment condition of RWC, the optimum composition of culture medium, and the optimum culture condition were examined. For sporulation, yeast cells were grown in the pre sporulation medium (PSM), transferred into sporulation medium (SM) containing 1 % potassium acetate, and incubated in a rotary shaking incubator at $25^{\circ}C$ for 4 days. The supernatant of the mixture of RWC and water was used as the presporulation medium (PSM). The optimum temperature and time for the pre-incubation of the mixture of RWC and water (1:2) to obtain maximum sporulation yield were $V^{\circ}C$ and 24 hr, respectively, and optimum culture time in PSM was 48 hr. Using these optimum conditions, the asci number obtained was 0.72$ 1.06${\times}$10^{8}$$m\ell$. The addition of wheat coat koji into SM increased the final number of asci to beTEX>$10^{8}$ $m\ell$. Spores were formed in the SM with the initial pH of 7-11, but no spores were formed in the SM with the initial pH of 5. To save the time and effort to pretreat the RWC, 2% and 0.5% RWC without any pretreatment were directly added into PSM containing 1 % brown sugar and SM, respectively, and the maximum asci number of $1.27${\times}$10^{8}$ /$m\ell$ was obtained.
Kim, Kyung-Hwan;Kim, Hea-Young;Ahn, Young-Soo;Lee, Woo-Choo;Hong, Sa-Suk
The Korean Journal of Pharmacology
/
v.20
no.2
/
pp.49-57
/
1984
The exocrine pancreatic secretion is controlled mainly by gastrointestinal hormones as well as cholinergic nerves. The adrenergic influence on exocrine pancreas is thought not to he important and the evidences supporting this contention are still contradictory. In an effort to elucidate the adrenergic influence on the exocrine pancreas, we have determined the amylase release from pancreatic slices of rats treated with adrenergic drugs. The albino rats of either sex, weighing $60{\sim}80\;g$, were decapitated and the uncinate pancreata were isolated and incubated in screw top vials containing 2 ml krebs-Ringer bicarbonate buffer solution gassed with 95% $O_2$ and 5% $CO_2$. These vials were shaken continuously in a waterbath maintained at $37^{circ}C$, and enzyme release was stimulated with acetylcholine$(10^{-5}M)$. For chronic treatment methoxamine$(an\;{\alpha}-adrenergic\;agonist,\;5\;mg/kg)$, isoproterenol (a\;{\beta}-adrenergic\;agonist,\;10\;mg/kg) and reserpine (0.5 mg/kg) along with cholecystokinin octapeptide$(CCK-op,\;2{\mu}g/kg)$ were given i.p. in rats daily for 3, 5, 7, 9 or 12 days. For acute experiment these drugs were added directly to the incubation medium in a concentration of $10^{-5}M$ except CCK-OP $(10^{-9}M)$. The results are summarized as follows. 1) The addition of methoxamine, isoproterenol or reserpine to the incubation medium containing pancreatic slices augmented the release of amylase induced by acetylcholine and among them the effect of isoproterenol was most prominent. 2) Chronic treatment of methoxamine or reserpine caused enhancement of acetylcholine response in amylase release from pancreatic slice throughout the experimental period, but the amylase release was less than that of control by 12 days isoproterenol treatment. 3) In the pancreatic slices obtained from 12 days treatment of CCK-OP, the amylae release responding to acetylcholine was enhanced. By these finding it is suggested that methoxamine, isoproterenol and reserpine had marked influence on the exocrine pancreatic functions in rats and that these effects are due to their inherent actions rather than sympathetic nerve or adrenergic receptor function.
Objectives: The object of this study was to observe the in vitro antibacterial effects of Chungdae-tang aqueous extracts, traditionally used for treating various gynecological diseases including vaginitis in Korea against Gardnerella vaginalis, and combination effects of Chungdae-tang extracts with Clindamycin were also monitored in this study. Methods: Antibacterial activities against Gardnerella vaginalis of Chungdae-tang aqueous extracts were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at MIC and MIC${\times}$2 levels. The combination effects of Chungdae-tang aqueous extracts with Clindamycin were observed by Checkerboard microtiter assay, and the effects of bacterial growth curve treated with or Chungdae-tang aqueous extracts MIC+Clindamycin MIC, 1/2MIC and 1/4MIC, respectively. In the present study, Gardnerella vaginalis were incubated under $37^{\circ}C$, 10% $CO_2$; and bacterial growth curves were calculated at 24, 48, 72, 96 and 120hrs after incubations. Results: MIC of Chungdae-tang aqueous extracts against Gardnerella vaginalis were detected as $3.906{\pm}2.344$(0.782~6.250) mg/$m\ell$, respectively. MIC of Clindamycin was detected as $0.010{\pm}0.006$(0.004~0.016) ${\mu}g/m\ell$ at same conditions. In addition, Clindamycin and Chungdae-tang aqueous extracts also showed marked dosage-dependent inhibition of bacterial growth, and more dramatical inhibitions were detected in Clindamycin+Chungdae-tang aqueous extracts MIC treatment as compared with each of single Clindamycin MIC and Chungdae-tang aqueous extracts MIC treatments, respectively. In addition, quite similar inhibitory effects on bacterial growth were detected in Clindamycin 1/4 MIC+Chungdae-tang aqueous extracts MIC treatment as compared with single Clindamycin MIC treatment in the present study. FIC index in combination of Chungdae-tang and Clindamycin were detected as $0.775{\pm}0.285$ (0.500~1.250) at Checkerboard microtiter assay. Conclusions: The results obtained in this study suggest that Chungdae-tang aqueous extracts showed antibacterial effects against Gardnerella vaginalis, and it also showed dosage-dependent inhibitory effects on the bacterial growth. In addition, combination treatment of Chungdae-tang aqueous extract with Clindamycin showed more potent inhibitory effects on the growth of Gardnerella vaginalis with FIC index $0.775{\pm}0.285$(0.500~1.250), respectively. It means, the combination of Chungdae-tang aqueous extract with Clindamycin is partially synergistic effects. It, therefore, is expected that effective dosages of Clindamycin will be reduced to 1/4 or over 1/4 levels as combination with Chungdae-tang extracts, respectively.
Park, Byung-Chan;Kim, Yong-Ha;Kim, Tae-Gon;LeeYoun-Jung, Jun-Ho;Sik-Young, Kim;Choi, Sik-Young
Archives of Plastic Surgery
/
v.37
no.4
/
pp.340-345
/
2010
Purpose: Recently, bioceramics have become popular as a substitute graft material for reconstruction of bony defect after trauma or tumor surgery. Among the bioceramic materials, hydroxyapatite (HA) is favored due to its biocompatibility. HA scaffold is composed of the interconnected reticular framework, macropores and micropores. Macropores play an important role in cell migration, nutrients supply and vascular ingrowth. On the other hand, a number of micropores less than $10{\mu}m$ form an irregular surface on HA scaffolds, which prevents the osteoblast from adhering and proliferating on the surface of HA scaffold. Methods: In this study, three different groups were designed for comparison. In the first group (group A), conventional method was used, in which HA pellet was applied without surface pretreatment. The second group (group B) was given a HA pellet that has been coated with crystalline HA solution prior to application. In the third group (group C), the same method was used as the second group, where the pretreated HA pellet was heated ($1250^{\circ}C$, 1 hour) before application. Osteoblast-like cells ($2{\times}10^4$/mL) were scattered onto every pellet, then they were incubated in 5% $CO_2$ incubator at $37^{\circ}C$ for twelve days. During the first three days, osteoblast cells were counted using the hemocytometer daily. ALP activity was measured on the 3, 6, 9 and 12 culture days using the spectrophotometer. Results: Under SEM, group A showed a surface with numerous micropores, and group B revealed more rough crystal surface. Group C revealed a fused crystal appearance and flattened smooth surface. In proliferation and ALP activity of osteoblast cells, group C showed better results compared to group B. Group A which lacks pretreatment of the surface showed less osteoblast proliferation and ALP activity than group C, but showed better results than group B. Conclusion: We found that crystallized HA with heat treatment method enhances the osteoblasts proliferation and differentiation on the surface of HA pellets.
The objective of this research was to examine the effect of ammonium thiosulfate(ATS) on urease activity and on biological and chemical properties of flooded paddy soil especially having high organic matter content by comparing with the effect of sodium thiosulfate(STS). The results obtained are summarized as follows. 1. The hydrolysis of urea was inhibited at 3 and 5 days after treatment of thiosulfate(ATS and STS) +glucose and thiosulfate only, respectively. The inhibitory effect of ATS on urea hydrolysis was slightly lower than that of STS in glusoce-added soils, but when the glucose was not added, the effects of ATS and STS were not different significantly. 2. The soil pH and Eh was lowered by 0.3~0.5 units and 30~120 mV, respectively, when incubated flooded soil with ATS and glucose at $25^{\circ}C$. 3. Soil respiration rate in/flooded soil was increased by 10~70% with the treatment of ATS during the 20 day experimental period. 4. The contents of acetic and butyric acid in thiosulfate treatment soil was below $10{\mu}g/g$, which was lower than that($220.3{\mu}g/g$) of critical growth inhibition of rice.
This study was to observe the changes of blastogenic responses of splenic Iymphocytes to T-cell mitogens, N. fcwleri Iysate and concanaualin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fcwleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with $7{\times}10^4$ trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000g for 60 minutes, and filtered through $0.2{\mu\textrm{m}}$ filter membrane. The supernatant, N. fcwleri Iysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred ${\mi}l$ of $10^6$ splenocytes in RPMI 1640 containing 0% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, $100{\mu}g/ml$ of N. fowleri Iysate or $4{\mu}g/ml$ of con. A, and the plates were incubated for 42 hours at $37^{\circ}C$ in 5% $CO_2$ incubator. Cultures were pulsed with of $methyl-(^3H)-thymidine$ 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fewleri Iysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri Iysate during the acute course of experimental PAM in mice.
The present study was undertaken to assess the role of cytokines in the activation of peritoneal macrophages from Toxoplasma-infected mice. Peritoneal macrophages from Toxoplasma-infected mice (10 cysts of Beverley strain/mouse) were harvested 8 weeks after infection, and incubated with the mitogen-induced lymphokine, recombinant mouse $interferon-{\gamma}(IFN-{\gamma})$, recombinant mouse tumor necrosis $factor-{\alpha}{\;}(TNF-{\alpha})$ alone or in combination with 4$IFN-{\gamma}(IFN-{\gamma}/TNF-{\alpha})$ for 24hr at 37^{\circ}C$, 5% $CO_2$. Macrophage activation was measured by the amount of $H_20_2{\;}and{\;}N0_2^{-}$ production, and antiToxoplasma activities of macrophages. $IFN-{\gamma}{\;}or{\;}IFN-{\gamma}/TNF-{\alpha}-treated$ macrophages from Toxoplasma-infected mice revealed significantly higher $H_20_2$ production than resident macrophages from Toxoplasma-infected mice. The production of $N0_2^{-}{\;}by{\;}TNF-{\alpha}-,{\;}IFN-{\gamma}-{\;}or{\;}IFN-{\gamma}/TNF-{\alpha}-treated$ macrophages from Toxoplasma-infected mice were significantly higher than that by resident macrophages, whereas lymphokine-treated group produced similar amount as that produced by resident macrophages. Anti-Toxoplasma activities of cytokinetreated macrophages from Toxoplasma-infected mice were Significantly higher than those of resident macrophages. $IFN-{\gamma}-treated$ macrophages were significantly increased production of $H_20_2{\;}and{\;}N0_2^{-}$, and anti-Toxoplasma activities of macrophages between normal and Toxoplasma-infected mice, whereas the other cytokine-treated groups were not significant differences between them. These data suggested that IFN-{\gamma}was the only one of cytokines capable of significantly activating the peritoneal macrophages from Toxoplasmainfected mice.
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