Eum, Soo Jin;Han, Seung Kyu;Chun, Kyung Wook;Kim, Woo Kyung
Archives of Plastic Surgery
/
v.36
no.1
/
pp.19-23
/
2009
Purpose: Platelet transplantation is a novel therapeutic strategy for acceleration of wound healing. When applying platelets, efficacy of adding thrombin to stimulate growth factor release from platelets has already been proved. However, no quantitative data of the thrombin treatment has been reported. The purpose of this study was to determine the optimal thrombin concentration to maximize growth factor release of platelets. In particular, this study was designed to quantify levels of platelet derived growth factor(PDGF)-BB, which is a major growth factor contained in the platelets, in vitro. Methods: Fresh platelets were obtained from a blood bank. They were suspended in DMEM/F - 12 and incubated with thrombin of various concentrations. The concentrations of thrombin tested were 0, 6.25, 12.5, 25, 50, 100, 200, and 400 IU/ml. After 30 minutes, 1, 3, 5, and 7 days, the levels of PDGF - BB were measured using enzyme linked immunosorbent assay. Platelets from four donors were included in this study. Each sample was tested in triplicate and the mean value was used as a data for each sample. Results: The addition of thrombin increased the level of PDGF - BB. Increases in storage time of platelets resulted in decreased levels of PDGF - BB. Higher levels of PDGF were detected in consort with increased thrombin concentrations. However, there was no significant difference between samples of 200 and 400 IU/ml concentrations. Conclusion: The results indicate that adding thrombin accelerates the release of growth factors from platelets and the optimal thrombin concentration to maximize this function is 200 IU/ml.
An experiment was conducted to establish a large scale production method of anti-serum against chicken IgM and to profile the developmental changes of serum IgM levels during the feeding period(from hatching to 7 weeks of age) in broiler chicks. Blood samples were taken from Hubbard chicken at the age of hatching, three days of age, and weekly thereafter till to 7 weeks of age. The pure IgM was isolated from ammonium sulfate treated chicken serum by both sephadex G-200 and sepharose CL-6B chromatography. The breaking-through peak containing IgM appeared from the fraction 26 to 28. These fractions consisted mainly of IgM when tested by anti-chicken IgM(Nordic, Netherlands). Immunized with the heavy chain of this purified IgM, the rabbit immune sera(anti-chicken IgM) were formed a reaction only with the purified chicken IgM. The quantitative assay of serum IgM were carried by RID method. The optimal time for diffusion was 14 hours and the coefficient of determination($R^{2}$) for regression equation of standard curve was 0.992. It was observed that IgM concentrations were the highest at hatching(3.23 mg /mL), after that decreased gradually. From 2 to 5 weeks of age the levels unchanged(2.0 ~ 2.3mg /mL), and gradually decreased to 7 weeks of age(1.3 mg /mL).
Kim, Dae-Sung;Jeon, Byoung-Kook;Mun, Yeun-Ja;Kim, Yeong-Mok;Lee, Young-Eun;Woo, Won-Hong
Journal of Physiology & Pathology in Korean Medicine
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v.25
no.3
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pp.425-430
/
2011
This study evaluated the effect of extraction conditions of yam (Dioscorea aimadoimo) on antioxidant, moisturizing, collagenase activity, proliferation, and migration. Yam has been recognized as a healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-mutagenic activities, as well as its ability to decrease blood glucose and cholesterol levels. Electron donating ability of high temperature ethanol extract of Dioscorea aimadoimo (HDA) had shown 70.6% at 400 mg/ml, and low temperature ethanol extract of Dioscorea aimadoimo (LDA) had shown 40% at 400 mg/ml. SOD-like activities of LDA and HDA were 23% and 34% at 400 mg/ml respectively. LDA significantly reduced the activity of collagenase in a dose-dependent manner, which was higher than HDA. The water contents in LDA-treated skin and HDA-treated skin were increased by 45.63% and 38.65% than the placebo cream respectively. The cellular proliferation of human dermal fibroblast neonatal (HDFn) was evaluated by MTT and cell migration assay. Compared to control, the cell proliferation was elevated to 109.7% and 114% by the treatment of LDA and HDA respectively at the concentration of 200 mg/ml. In addition, LDA and HDA were induced cell migration in HDFn. Our study suggests that LDA and HDA should be a very useful cosmetic ingredient, as anti-aging and skin moisturizer.
3-Monochloro-1,2-propanediol(3-MCPD) is a toxic compound, often present in different foods containing acid hydrolyzed(AH) protein, like seasonings and savory food products. The purpose of the present study was to investigate the effects of 3-MCPD on male fertility, sperm and testosterone secretion. In vivo male fertility test was performed for observing the adverse effects of 3-MCPD on the function of male reproductive system and pregnancy outcome. 0.01, 0.05, 0.25, 1 and 5 mg/kg b.w. of 3-MCPD was given daily by gavage to groups of 15 adult male SD rats for 4 weeks. At the end of pre-treatment period, males were mated overnight with normal females. Following morning, males demonstrating successful induction of pregnancy were sacrificed on that day to assess sperm parameters and histopathology of reproductive organs. The resulting pregnant females were sacrificed on day 20 of gestation to evaluate pregnancy outcome. As a result, four-week paternal administration with 3-MCPD resulted in adverse effects on male fertility and pregnancy outcome without remarkable histopathological changes in testes and epididymides; sperm motility, copulation index and fertility index were markedly decreased in the treated group and numbers of live fetuses showed steep dose-response curves. Also, spermatogenesis was investigated in this experiment. However, no effect was observed on production of sperm in testes treated with 3-MCPD for 4 weeks. Hormone assay was performed for observing the effects of 3-MCPD on testosterone and luteinizing hormone (LH) in blood and testes of male SD rats and cultured primary Leydig cell. In result, significant changes of related hormones did not observed by treatment of 3-MCPD. These results indicated that paternal treatment with 3-MCPD induced spermatotoxic effect, which caused an antifertility on male.
We had isolated 8 viruses from 91 specimen collected at southwest Cheju Province during early spring 1998 and 2 viruses from 9 specimen collected at Chung Nam Province during early spring 1999. To perform cross-reactivity among 4 mumps vaccine strains and 10 wild-type mumps viruse isolates, we immunized mice and took antisera against each virus. There were no antibody titer differences by indirect immunofluorescence assay (IFA), but most isolated mumps viruses showed a little cross-reactivities with Jeryl Lynn and Rubini strains. It has shown similar result by haemagglutination-inhibition (HAI) test. These results show that 4 mumps strains used as vaccine have the protection ability against endemic wild-type mumps viruses. Also the SH gene analysis was performed to identify genotypes. Most isolated mumps viruses belonged to genotype D. These results indicate that endemic mumps viruses in Korea are different to ones isolated in Japan and China.
Eye is a highly vascularised organ. There are chances that a foreign substance can enter the systemic circulation through the eye and cause oxidative stress and evoke immune response. Here the eyes of rabbits were exposed, for a period of 7 days, to 5 known ocular irritants: Cetyl pyridinium chloride (CPC), sodium salicylate (SS), imidazole (IMI), acetaminophen (ACT) and nicotinamide (NIC). The eyes were scored according to the draize scoring. Blood collected from the treated rabbit were analyzed for haematological and biochemical parameters. After sacrifice, histological analysis of the eye and analysis of pro-inflammatory biomarkers ($IL-1{\alpha}$, $IL-1{\beta}$, IL-8 and $TNF-{\alpha}$) in the cornea using ELISA was carried out. Spleen was collected and the proliferation capacities of spleenocytes were analyzed. Liver and brain were collected and assessed for oxidative stress. The eye irritation potential of the chemicals was evident from the redness and swelling of the conjunctiva and cornea. Histopathological analysis and ELISA assay showed signs of inflammation in the eye. However, the haematological and biochemical parameters showed no change. Spleenocyte proliferations showed only slight alterations which were not significant. Also oxidative stress in the brain and liver were negligible. In conclusion, chemicals which cause ocular irritation and inflammation did not show any systemic side-effects in the present scenario.
The pathogenesis and pathophysiology of Acanthamoeba infections remain incompletely understood. Phospholipases are known to cleave phospholipids, suggesting their possible involvement in the host cell plasma membrane disruption leading to host cell penetration and lysis. The aims of the present study were to determine phospholipase activities in Acanthamoeba and to determine their roles in the pathogenesis of Acanthamoeba. Using an encephalitis isolate (T1 genotype), a keratitis isolate (T4 genotype), and an environmental isolate (T7 genotype), we demonstrated that Acanthamoeba exhibited phospholipase $A_2$ (PLA$_2$). and phospholipase D (PLD) activities in a spectrophotometry-based assay. Interestingly, the encephalitis isolates of Acanthamoeba exhibited higher phospholipase activities as compared with the keratitis isolates, but the environmental isolates exhibited the highest phospholipase activities. Moreover, Acanthamoeba isolates exhibited higher PLD activities compared with the PLA$_2$. Acanthamoeba exhibited optimal phospholipase activities at $37^{\circ}C$ and at neutral pH indicating their physiological relevance. The functional role of phospholipases was determined by in vitro assays using human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. We observed that a PLD-specific inhibitor, i.e., compound 48/80, partially inhibited Acanthamoeba encephalitis isolate cytotoxicity of the host cells, while PLA$_2$-specific inhibitor, i.e., cytidine 5'-diphosphocholine, had no effect on parasite-mediated HBMEC cytotoxicity. Overall, the T7 exhibited higher phospholipase activities as compared to the T4. In contract, the T7 exhibited minimal binding to, or cytotoxicity of, HBMEC.
To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.
Kim, Byung-Seok;Lim, Hyun-Sul;Lee, Kwan;Min, Young-Sun;Yoon, Young-Sil;Jeong, Hye-Sook
Journal of Preventive Medicine and Public Health
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v.48
no.1
/
pp.53-61
/
2015
Objectives: The seroprevalence of hepatitis E virus (HEV) among high-risk groups overseas is high, but studies in these groups are rare in South Korea. We conducted the present study from April to November 2012 to obtain data on the seroprevalence and associated risk factors for HEV among slaughterhouse workers in South Korea. Methods: Slaughterhouse workers from 80 workplaces nationwide were surveyed in South Korea in 2012. The subjects comprised 1848 cases: 1434 slaughter workers and 414 residual products handlers. By visiting 80 slaughterhouses, which were mixed with 75 of which also performed residual products handling, we conducted a questionnaire survey for risk factors and obtained blood samples in order to determine the seropositivity and seroprevalence of HEV. Anti-HEV IgG and IgM were measured using HEV IgG and IgM enzyme-linked immunospecific assay kits and HEV antigen was measured by reverse transcription polymerase chain reaction (RT-PCR). Results: The seropositivity of anti-HEV IgG was 33.5% (slaughter workers 32.8% and residual products handlers 36.2%), and among the seropositive individuals the seroprevalence of anti-HEV IgM was 0.5% (slaughter workers 0.5%, residual products handlers 0.7%). The response rate of HEV-antigen as measured by RT-PCR was 0.2%. Risk factors significantly related to anti-HEV IgG seropositivity were age, sex, and working duration (slaughter workers only). Conclusions: There were significant risk factors (sex, age, and working duration) for HEV identified in our study. All three positive cases for HEV-antigen by RT-PCR were related to pig slaughter but without statistical significance. To prevent HEV, an educational program and working guidelines may be needed for high risk groups.
Park, Hee-Juhn;Nam, Jung-Hwan;Jung, Hyun-Ju;Lee, Myung-Sun;Lee, Kyung-Tae;Jung, Min-Hwa;Choi, Jong-Won
Korean Journal of Pharmacognosy
/
v.36
no.4
s.143
/
pp.324-331
/
2005
The roots of Rosa rugosa have been used to treat diabetes mellitus in the folkloric society of Korea. To demonstrate the active component for the rat obesity induced by high fat diet for 6 weeks, the phytochemical fractionation and the pharmacological activity test were performed on this crude drug. It was shown that the methanolic extract and its EtOAc fraction inhibited the weight increase of the rat body, abdominal fat pad and hyperlipidemia at 200 mg/kg dose. Further, the triterpenoids, euscaphic acid and tormentic acid, isolated from R. rugosa roots were active at 30 mg/kg in the same assay. The two components shifted serum total-, HDL, and LDL-cholesterol levels toward the values of the unteated group, suggesting that the active compounds has hypolipidemic effects. The rats fad euscaphic acid and tormentic acid also reduced thiobarbituric acid-reactive substance (TBARS) and hydroxyl radical in the rat blood and increased superoxide dismutase activity compared to the control. TBARS values and carbonyl contest of the hepatic protein were reduced by treatment with the two triterpenoids. Antioxidative enzyme (SOD, glutathione peroxidase, and catalase) activities in hepatic were increased by treatment of rats with the triterpenoids, which suggests that triterpenoids inhibited the reduction of hepatic antioxidative activity caused by high fat diet. Taken together, these results support that euscaphic acid and tormentic acid improve a high fat diet-induced hyperlipidemia via the activation of antioxidative mechanism.
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