Purpose : We hypothesized dexrazoxane(DXR) and pentoxifylline(PTX) may prevent myocardial damage in adriamycin(ADR)-induced cardiomyopathic rat model. We also investigated their effects on the myocardial apoptosis and fibrosis in ADR induced cardiomyopathy. Methods : The six-week old female Spregue-Dawley rats were divided into control group(CNT, n=4), ADR group(n=6), ADR+DXR group(DXR, n=5), ADR+PTX group(PTX, n=6), ADR+DXR+PTX group(DXPT, n=5). ADR(5 mg/week, twice) was administrated intravenously to rats except CNT group to induce cardiomyopathy. The PTX(50 mg/kg/day) was administered daily from day-0 to Day-21. The DXR(100 mg/kg) was administered 30 minutes before each ADR injection. On day 21, the rats were sacrificed and the degree of histopathologic changes of hypercontraction band necrosis, cytoplasmic vacuolar change and fibrosis were scored. Immunohistochemical staining for Bcl-2 expression and RT-PCR for $TNF-{\alpha}$ and CTGF were performed. Results : Histopathological scores of myocardial damage were significantly higher in ADR rats than CNT rats(P<0.05), and significantly lower in DXPT rats than ADR rats(P<0.01). Myocardial fibrosis was prevented in both PTX rats and DXPT rats. The expression of Bcl-2 was weaker in ADR rats than that in CNT rats(P<0.05), and stronger in both DXR and DXPT rats than that in ADR rats (P<0.05). $TNF-{\alpha}$ concentration of ADR rats was not different from that of treated groups. Conclusion : DXR prevented myocyte apoptosis with increased Bcl-2 expression, and PTX prevented myocardial fibrosis in ADR induced cardiomyopathic rats. The combination therapy of DXR and PTX showed prevention of cardiomyopathy in ADR induced cardiomyopathy rat model.
Purpose : Human umbilical vein endothelial cells(HUVECs) play an important role in regulating blood flow by releasing vasoactive substances. It has been reported that endothelial impairment and dysfunction might be a primary cause of placental vascular disease, which is manifested clinically as preeclampsia in mother and intrauterine growth restriction in fetus. Furthermore, the frequency of apoptotic changes is increased in umbilical and placental tissues from growth-restricted pregnancies. However, the various mechanisms of umbilical endothelial cell apoptosis have not been broadly proposed. We investigate the effects of amiloride derivatives on apoptotic death of HUVECs and identify their ionic mechanism. Methods : HUVECs were purchased from Clonetics, and cultured on endothelial cell growth medium. MTT assay and flow cytometry were used for assessing cytotoxic effect and confirming the apoptosis. Changes in intracellular ion concentrations were measured with specific fluorescent dyes and fluorescence imaging analysis system. Results : Amiloride derivatives elicited cytotoxic effects on HUVECs with dose-dependent manners and the rank order of potency is HMA($IC_{50}\;11.2{\mu}M$), MIA>EIPA>>amiloride. HMA-induced cytotoxicity is dependent on extra- and intracellular pH, that is, increase extra- and intracellular pH augmented the cytotoxic effects of HMA. HMA dose-dependently reduced intracellular major ions, such as $K^+$ and $Cl^-$. Interestingly, the depletion of intracellular ions induced by HMA was also significantly enhanced at alkaline extracellular pH. Conclusion : Amiloride derivatives induce apoptosis of HUVECs with dose and pH dependent manners. They reduce intracellular $K^+$ and $Cl^-$ concentration, which is also extracellular pH dependent.
This study has been carried out in order to investigate the bacterial quality of fish meat paste products and the characteristics of isolated thermodurics from the products. Twenty samples of crab-flavored fish stick (Kematsal), 23 samples of plate fish meat paste (Panomuk, Kamaboko), 5 samples of fried fish meat paste (Tigimomuk), 2 samples of roasted fish meat paste (Puduromuk, Chikuwa), 20 samples of fish sausage were collected from processing plants and supermarkets in Pusan, Korea during the period from May to October in 1984. The results obtained are as follows. Amont the samples collected from supermarkets, roasted fish meat paste and fried fish meat paste marked hish counts in coliforms and fungi while very low in the samples of crab-flavored fish stick and plate fish meat paste. Salmonella was not detected in all the samples examined and Staphylococcus aureus was detected only in fried fish meat paste, Thermoduric bacteria were detected less than 10$^2$/g in the samples of crab-flavored fish stick and plate fish meat paste, which might come from subsidiary materials such as starch and seasonings. Among the isolated bacteria, distribution of the proteolytics were more than 87% and the lipolytics were less than 20%. Gram positive bacteria was more than 70% in crab-flavored fish stick and plate fish meat paste, 47.3% in fried fish meat paste. And rod in shape was almost more than 90% in all the samples. The most heat resistant bacterium isolated from the samples was identified as a Bacillus licheniformis(named B. licheniformis CR-11). The strain showed strong proteolytic activity and also grew well at above 2$0^{\circ}C$. The growth rate and generation time of CR-11 strain were 0.31 hr$^{-1}$ , 2.24 hr at 2$0^{\circ}C$, 0.64 hr$^{-1}$ , 1.09 hr at 3$0^{\circ}C$ and 0.78 hr$^{-1}$ , 0.89 hr at 35$^{\circ}C$. Heat resistance value of the spores of CR-11 strain suspended in phosphate buffer solution was D$_{85}$$^{\circ}C$=41.9 min, D$_{90}$$^{\circ}C$=27.9 min, D$_{95}$$^{\circ}C$=10.2 min, D$_{100}$$^{\circ}C$=4.3 min (Z=13.8$^{\circ}C$)
An, Na Young;Kim, Ji-Eun;Hwang, DaeYoun;Ryu, Ho Kyung
Journal of Nutrition and Health
/
v.47
no.6
/
pp.394-402
/
2014
Purpose: Dendropanax morifera Leveille (DML) exhibits diverse biological and pharmacological activities, including anti-oxidative effect, anti-cancer activity, hepatoprotection, immunological stimulation, and bone regeneration. As part of the identification for novel functions of DML, we investigated the therapeutic effects of DML on diabetes induced by streptozotocine (STZ) treatment. Methods: First, the four extracts including the water extract of leaf (DLW), the ethanol extract of leaf (DLE), the water extract of stem (DSW), and the ethanol extract of stem (DSE) were collected from the leaf and stem of DML using a hot water and ethanol solvent. Alterations in body weight, glucose concentration, insulin level, and pancreatic islet structure were investigated in diabetic mice after treatment with extracts of DML for 2 weeks. Results: Among four extracts, the highest level of total polyphenols and total flavonoids was detected in DLW, while the lowest level of these was measured in DSE. The radical scavenging activity was also higher in DLW than in the other three extracts at the concentration of $25-100{\mu}g/mL$, although this activity was maintained at a constant level in all groups at the concentration of $500{\mu}g/mL$. Based on the results of anti-oxidant activity, DLW and DLE were selected for examination of anti-diabetic effects in a diabetes model. Body weight was gradually decreased in all STZ treated groups compared with the No treated group. However, four STZ/DML treated groups maintained a high level of body weight during 7-14 days, while the STZ/vehicle treated group showed a gradual decrease of body weight during the same period. Also, a significant decrease or increase in the concentration of glucose and insulin in the blood of the diabetes model was detected in a subset of groups, although the highest increase was detected in the STZ/DLE-200 treated group. In addition, the histological structure of pancreatic islet was significantly recovered after treatment with DLW and DLE. Conclusion: These results suggest that DLW and DLE may contribute to attenuation of clinical symptoms of diabetes as well as prevent the destruction of pancreatic ${\beta}$-cells in STZ-induced diabetes mice.
Kim, Tae Woo;Kim, Kyoung Kon;Kang, Yun Hwan;Kim, Dae Jung;Choe, Myeon
Journal of Nutrition and Health
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v.47
no.4
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pp.221-228
/
2014
Purpose: Citron seed oil (CSO) has been reported to have high antioxidant activity. However, the composition and other biologically activities of CSO have not been reported. In this study, we confirmed the fatty acid composition of CSO, which may be beneficial to vascular disease and obesity. Methods: We investigated the oil composition of CSO using gas chromatography coupled with mass spectrometry (GC-MS) analysis, and cytotoxicity was confirmed by Cell Counting Kit-8 (CCK-8) assay. Nitric oxide (NO) production in human umbilical vein endothelial cells (HUVECs) was measured using Griess reagent, and lipid accumulation and leptin secretion in 3T3-L1 cells were measured by Oil-Red O staining and commercial ELISA kit, respectively. Results: GC-MS analysis indicated that CSO contains several components, including linoleic acid, oleic acid, palmitic acid, stearic acid, linolenic acid, palmitoleic acid, and arachidic acid. In physiological activity analysis, CSO did not induce cytotoxic effects in HUVECs and 3T3-L1 cells. Further, CSO significantly induced nitric oxide and leptin secretion as well as inhibited lipid accumulation. Conclusion: CSO increased NO release, inhibited lipid accumulation, and induced leptin secretion, suggesting it may be useful for the management of vessels and weight gain. Although further studies are required to investigate the safety and mechanism of action of CSO, our results show that the composition and physiological activity of CSO are sufficient for its use as functional edible oil.
Bone resorptiion was predominate in compression site, bone formation in tension site of periodontal ligament during tooth movement. The biologic response at compressiion site was different from tension site. Thus the CGHP immuno-positive nerve fiber will respond differently to mechanical force according to the area( compression or tension site). The purpose of this study was to investigate this response of CGRP immune-positive nerve fiber in the periodontal ligament according to the duration of applied force and the area (compression or tension site) during experimental tooth movement. The experimental animals were 7 week old male rat (approximately 200 gm). The orthodontic force was applied mesially to the right maxillary molar using the Ni-Ti coil spring during 12hours, 1, 3, 7, and 120days. Immunohistochemical staining using antibody against CGRP was performed after sacrifice. The results were as follows. The CGRP immune-positive nerve bundle showed reduced immunoreactivity and nerve fibers reduced in density after application of orthodontic force for 12 hours and 1day. The CGRP immune-positive nerve fibers showed many thin branches at the apical periodontal ligament after application of force for 3 days as compared with normal. The tension site in the apical periodontal ligament showed more branches than the compression site. In 7 day group, the CGRP immune-positive nerve fibers increased in terms of the number and had many thin branches in the apical periodontal ligament. The tension site had more branches than the compression site. In 12 day group, the CGRP immune-positive nerve fibers showed similar distribution to normal control at compression site of apical periodontal ligament, but the fibers at the tension site increased in number. The CGRP immuno-positive nerve fibers showed more increased at tension site than compression site after application of orthodontic force. Therefore it seems to have some relation to the bone remodeling besides the local inflammatory process.
Conventional osseointegrated titanium implants have many limitations; large size, limited location for placement of the implant, severity of the surgery, discomfort of initial healing, difficulty of oral hygiene and uncontrollable force direction. Recently titanium miniscrew and miniplate have been used for an alternative to conventional dental implant. But in relation to miniplate, miniscrew has disadvantages in that more potential inflammation, light orthodontic force application and limited orthodontic application. This study was conducted to evaluate the effectiveness of miniplate by observing the reactions of peri-implant tissues to early orthodontic and orthopedic loading of titanium miniplate. In four adult beagle dogs 10 miniplates were inserted into the alveolar bone using 20 osseointegrated titanium screws. 4 miniplates were placed in two dogs(dogA, B) and 6 miniplates in two dogs(dogC, D). In dogA, B miniplates were loaded with 200gm of force immediately after placement for 15 weeks. In dogc, D, miniplates were loaded with 400gm of force immediately after placement for 8 weeks. Miniplates of dogA were removed, dogA was healed for 4 weeks, and the area which was removed of miniscrew was examined. Following an observation period, the miniplates including miniscrews and the surrounding bone of dogB and dogC, D were removed, respectively. Undecalcified section along the long axis of miniscrews were made and the degree of osseointegration was examined under the light microscope. The results were as follows. 1. In the histologic features there was direct contact between bone and miniscrew in all groups except one, dogC control group. The loaded miniscrew demonstrated only a slight increase of the osseous proximaty when compared with unloaded miniscrew 2. There was no significant difference of the osseointegration of Peri-miniscrew surface between dogB and dogC, D. But dogB showed slightly more increased bone apposition than dogC, D 3. The gingiva overlapping the miniplate and miniscrew showed no inflammatory sign in clinical and histological aspects. 4. The impaled hard and soft tissues at the area which was removed of miniscrews showed good healing without inflammatory reaction. 5. The mobility showed slight increase in un-loaded miniplate but that was insignificant. Based on the results of this study, miniplate(C-tube) can be used as a firm osseous orthodontic and orthopedic anchorage unit immediately after insertion.
The aims of this study were to establish the effect of gender and age on number of fungiform papillae and electrogustometric thresholds of the tongue, and to determine the relation between number of fungiform papillae and electrogustometric threshold. Sixty-two healthy subjects of 20 to 39 years old were enrolled in this study. Examination areas were defined as center, anterior, left lateral, right lateral, and posterior on the dorsal surface of the tongue, which was stained with methylene blue dye and photographed with a scale bar on it. The number of fungiform papillae within $1\; cm^2$ square area of each examination site was counted twice. The counted values were averaged to produce means. Taste thresholds were measured three times on each examination area using an electrogustometer. Means of the threshold values were calculated. 1. The number of fungiform papillae was the largest at the anterior examination area, followed by right lateral, left lateral, posterior and central. Women had more fungiform papillae than men at anterior, right lateral, and center areas. Subjects in twenties showed more fungiform papillae than those in thirties at anterior, right lateral, and left lateral areas. 2. Anterior area of the tongue measured the lowest electrogustometric threshold, followed by left lateral, right lateral, posterior and center. The threshold was higher in men at anterior and right lateral areas. The subjects in thirties had higher threshold only at right lateral area than those in twenties. 3. Electrogustometric threshold was inversely correlated with the number of fungiform papillae at anterior and posterior areas. The results showed that the number of fungiform papillae and taste threshold varied according to tongue locus, and they were influenced by age and gender. The results also suggested that the number of fungiform papillae was related to electrogustometric threshold inversely.
Morphological and cultural characteristics of fungi causing rice sclerotial diseases were examined. Hyphal widths of R. solani and R. oryzae were same and ranged $6.0-12.0\;{\mu}m$ with average $9.0\;{\mu}m$, the widest among those of the sclerotial fungi examined. Hyphal width of R. oryzae sativae ranged $6.0-9.0{\mu}m$ with average $7.4{\mu}m$. Hyphal width of R. cerealis was the narrowest among those of Rhizoctonia species examined, and the same was hyphal width of S. oryzae among those of Sclerotium species. Nuclear staining by HCL-Giemsa method showed that R. solani and R. oryzae had many nuclei within one hyphal cell, S. oryzae one nucleus, and the other sclerotial fungi mostly two nuclei. The nuclear number of R. solani was the largest, which ranged 2-17 with average 6.3. Average size of sclerotia of the sclerotial fungi except S. hydrophilum and S. oryzae produced in lesions ranged 1.0-2.0mm. Average size of sclerotia of S. hydrophilum and S. oryzae was 0.5mm and 0.24mm, respectively. Sclerotia of R. solani and R. oryzae produced in culture were more variable in size and larger than those produced in lesions. However, the sclerotial sizes of the other sclerotial fungi produced in culture were almost the same as those produced in lesions. Sclerotial colors of sclerotial fungi produced in lesions were similar to those produced in culture, but sclerotial shapes of some sclerotial fungi exhibited somewhat difference between the sclerotia produced in lesions and in culture. Optimum temperature for mycelial growth of R. cerealis was $23^{\circ}C$, and that of the other sclerotial fungi ranged from $27\;to\;33^{\circ}C$. Maximum temperature for mycelial growth of some sclerotial fungi was as high as $41^{\circ}C$, while that of R. cerealis was as low as $31^{\circ}C$. Minimum temperature for mycelial growth of R. cerealis was $2^{\circ}C$, and that of the other sclerotial fungi ranged from $6\;to\;10^{\circ}C$.
The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa), group 8 (low-temperature preservation at $0^{\circ}C$ under no additional pressure), group 9 (low-temperature preservation at $-5^{\circ}C$ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 ($0^{\circ}C$/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
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