The molecular technique for 168 rRNA gene sequences analysis and Western Blot were applied for determination of the Bacillus probiotic strains used in biosubtyl production at IVAC. The results showed that the nucleotide sequence identity of the NT strains was 99.74% in comparison with B. pumilus and the identity of the DL strains was 99.61% in comparison with B. cereus strains from the GenBank/EMBL/DDBJ Database. The result of Western blot analysis showed that, the antigen components of the strains NT and DL were not similar in their antigens, and they were not similar to the B. subtilis ATCC6633.
Bacillus ; Blotting, Western

PURPOSE: To investigate the presence of adaptive response by low dose radiation in murine tumors in relation to radiation induced apoptosis as well as related mechanism. MATERIALS AND METHODS: Syngeneic murine tumors, OCa-I and HCa-I, were given 0.05 Gy pretreatment followed by therapeutic dose of 25 Gy radiation. Induction of apoptosis was analyzed for each treatment group. Regulating molecules of apoptosis, p53, Bcl-2, Bax, Bcl-X, were also analyzed by Western blotting. RESULTS: In 0.05 Gy pretreatment group of OCa-I, 25 Gy-induced apoptosis per 1000 cells was 229, which was estimated at 30% lower level than the expected (p<0.05). In contrast, this reduction in radiation induced apoptosis was not seen in HCa-I. In the expression of apoptosis regulating molecules, p53 increased in both tumors in response to radiation. Bcl-2 and Bax did not show significant change in both tumors however, the expression of Bcl-2 surpassed that of Bax in 0.05 Gy pretreatment group of OCa-I. Bcl-X was not expressed in OCa-I. In HCa-I, Bcl-X showed increased expression even with 0.05 Gy. CONCLUSION: Adaptive response by low dose radiation is shown in one murine tumor, OCa-I, in relation to radiation induced apoptosis. Apoptosis regulating molecules including Bcl-2/Bax and Bcl-X, appear to related. This study shows an evidence that adaptive response is present, but not a generalized phenomenon in vivo.
Apoptosis* ; Blotting, Western ; Radiation Dosage

BACKGROUND: Various kinds of hair products are widely used due to the increase of interest in hair styling. Cosmetic procedures such as permanent waving are very popular today, but the medical studies related to the meaning and restoration pattern of hair damage are mainly based on structural findings. In measuring the degree of hair damage by the moleculobiological methods rather than the structural studies, the findings seem to be highly objective and standardized. OBJECTIVE: The purpose of this study was to identify the patterns of hair damage and restoration through electrophoresis and western blot analysis of hair proteins. METHODS: The three volunteers who we selected as subjects did not have any specific medical illness and had not performed any special cosmetic procedures which could have caused hair damage during the six months before the study. We conducted permanent waving on them. Human hair samples were obtained from the occipital scalp, which were that was not affected by the androgen. We performed extraction and concentration of the whole and partial hair protein, then operated electrophoresis and western blot analysis of the hair protein. RESULTS: In the western blot analysis of whole hair proteins, there was one positive finding on subject A. This may have resulted from the small amount of partial proteins among the whole hair proteins. In the western blot analysis of partial hair proteins, subject A and B showed positive findings. In particular, positive findings were found on the 14th week of the experiment. CONCLUSION: These results show a change in the hair proteins due to hair damage, and ultrastructurally, we found the possibility of prologation of actual hair damage longer than expected.
Blotting, Western ; Electrophoresis ; Hair* ; Humans ; Scalp ; Volunteers

BACKGROUND: Sorbus rufopilosa, a tsema rowan, is a species of the small ornamental trees in the genus Sorbus and the family Rosaceae found in East Asia. The bioactivities of S. rufopilosa have not yet been fully determined. The objective of this study is to evaluate the antioxidant and anticancer effects of ethanol extract of S. rufopilosa (EESR) and to determine the molecular mechanism of its anticancer activity in human colon carcinoma HT29 cells. METHODS: To examine the antioxidant activity of EESR, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity assay was performed. Inhibitory effect of EESR on cancer cell growth and proliferation was determined by water-soluble tetrazolium salt assay. To investigate the mechanism of EESR-mediated cytotoxicity, HT29 cells were treated with various concentrations of EESR and the induction of cell cycle arrest and apoptosis was analyzed by flow cytometry, 4,6-diamidino-2-phenylindole staining, and Western blot analysis. RESULTS: EESR showed significant antioxidant activity and inhibitory effect on HT29 cell growth in a dose-dependent manner. EESR induced cell cycle arrest at G2/M phase in a dose-dependent manner by modulating cyclin B, cyclin-dependent kinase 1 (CDK1), and CDK inhibitor p21 expression. EESR-induced apoptosis was associated with the upregulation of p53, a death receptor Fas, and a pro-apoptotic protein Bax and the activation of caspase 3, 8, and 9, resulting in the degradation of PARP. CONCLUSIONS: EESR possessing antioxidant activity efficiently inhibits proliferation of HT29 cells by inducing both cell cycle arrest and apoptosis. EESR may be a possible candidate for the anticancer drug development.
Adenocarcinoma* ; Apoptosis* ; Blotting, Western ; Caspase 3 ; CDC2 Protein Kinase ; Cell Cycle Checkpoints* ; Cell Cycle* ; Colon* ; Cyclin B ; Ethanol ; Far East ; Flow Cytometry ; HT29 Cells* ; Humans* ; Rosacea ; Rosaceae ; Sorbus* ; Trees ; Up-Regulation

BACKGROUND: Asian dust storms can be transported across eastern Asia. In vitro, Asian dust particle-induced inflammation and enhancement of the allergic reaction have been observed. However, the fibrotic effects of Asian dust particles are not clear. Production of transforming growth factor beta1 (TGF-beta1) and fibronectin were investigated in the bronchial epithelial cells after exposure to Asian dust particulate matter (AD-PM10). METHODS: During Asian dust storm periods, air samples were collected. The bronchial epithelial cells were exposed to AD-PM10 with and without the antioxidant, N-acetyl-L-cysteine (NAC). Then TGF-beta1 and fibronectin were detected by Western blotting. The reactive oxygen species (ROS) was detected by the measurement of dicholorodihydrofluorescin (DCF), using a FACScan, and visualized by a confocal microscopy. RESULTS: The expression of TGF-beta1, fibronectin and ROS was high after being exposed to AD-PM10, compared to the control. NAC attenuated both TGF-beta1 and fibronectin expression in the AD-PM10-exposed the bronchial epithelial cells. CONCLUSION: AD-PM10 may have fibrotic potential in the bronchial epithelial cells and the possible mechanism is AD-PM10-induced intracellular ROS.
Acetylcysteine ; Air Pollutants ; Asian Continental Ancestry Group ; Blotting, Western ; Dust ; Epithelial Cells ; Far East ; Fibronectins ; Humans ; Hypersensitivity ; Inflammation ; Particulate Matter ; Reactive Oxygen Species ; Transforming Growth Factor beta ; Transforming Growth Factor beta1

Angelica koreana is an important medicinal plant for some locals in East Asia including Korea. A few reports have shown the efficacy of its phytochemical constituents. We have isolated and purified one compound falcarindiol (FAL) from the methanolic extract of A. koreana roots. At concentrations from to 1 µM to 25 µM, the FAL isolated from the roots of A. koreana exerted no significant cytotoxicity and down-regulated LPS-stimulated pro-inflammatory cytokine IL-8 in colon epithelial cells, while up-regulating anti-inflammatory cytokine IL-10. In addition, the FAL decreased the expression of LPS-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein by Western blot analysis. Colon epithelial cells play pivotal roles in regulating the colon immune system and thus FAL is expected to be candidate agent as therapeutic potential for the treatment of inflammatory bowel disease (IBD) by modulating LPS-induced inflammation in colon epithelial cells.
Angelica* ; Blotting, Western ; Colon* ; Epithelial Cells* ; Far East ; Immune System ; Inflammation ; Inflammatory Bowel Diseases ; Interleukin-10* ; Interleukin-8* ; Korea ; Methanol ; Nitric Oxide Synthase Type II ; Plants, Medicinal ; Prostaglandin-Endoperoxide Synthases

Shikonin, which derives from Lithospermum erythrorhizon, has been traditionally used against a variety of diseases, including cancer, in Eastern Asia. Here we determined that shikonin inhibits proliferation of gastric cancer cells by inducing apoptosis. Shikonin's biological activity was validated by observing cell viability, caspase 3 activity, reactive oxygen species (ROS) generation, and apoptotic marker expressions in AGS stomach cancer cells. The concentration range of shikonin was 35–250 nM with the incubation time of 6 h. Protein levels of Nrf2 and p53 were evaluated by western blotting and confirmed by real-time PCR. Our results revealed that shikonin induced the generation of ROS as well as caspase 3-dependent apoptosis. c-Jun-N-terminal kinases (JNK) activity was significantly elevated in shikonin-treated cells, thereby linking JNK to apoptosis. Furthermore, our results revealed that shikonin induced p53 expression but repressed Nrf2 expression. Moreover, our results suggested that there may be a co-regulation between p53 and Nrf2, in which transfection with siNrf2 induced the p53 expression. We demonstrated for the first time that shikonin activated cell apoptosis in AGS cells via caspase 3- and JNK-dependent pathways, as well as through the p53-Nrf2 mediated signal pathway. Our study validates in partly the contribution of shikonin as a new therapeutic approaches/ agent for cancer chemotherapy.
Apoptosis ; Blotting, Western ; Caspase 3 ; Cell Death* ; Cell Survival ; Drug Therapy ; Far East ; Humans* ; Lithospermum ; Phosphotransferases ; Reactive Oxygen Species ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Stomach Neoplasms ; Stomach* ; Transfection

PURPOSE: To analyze the involvement of apoptosis regulatory genes p53, p21waf1/cip1, bax and bcl-2 in induction of apoptosis by radiation in murine tumors. MATERIALS AND METHODS: The radiation-sensitive ovarian carcinoma OCa-I, and the radiation-resistant hepatocarcinoma HCa-I were used. Tumors, 8 mm in diameter, were irradiated with 25 Gy and at various times after irradiation, ranging from 1 to 48 h, were analyzed histologically for apoptosis and by western blot for alterations in the expression of these genes. The p53 status of the tumors were determined by the polymerase chain reaction-single strand conformation polymorphism assay. RESULTS: Both tumors were positive for wild-type p53. Radiation induced apoptosis in OCa-I but not in HCa-I. Apoptosis developed rapidly, peaked at 2 h after irradiation and returned to almost the background level at 48 h. In OCa-I radiation upregulated the expression of p53, p21waf1/cip1, and the bcl-2/bax ratio was decreased. In HCa-I radiation increased the expression of both p53 and p21waf1/cip1, although the increase of the latter was small. The bcl-2/bax ratio was greatly increased. In general the observed changes occurred within a few hours after irradiation, and either preceded or coincided with development of apoptosis. CONCLUSIONS: The development of apoptosis required upregulation of both p53 and p21waf1/cip1 as well as a decrease in bcl-2/bax ratio. In contrast, an increase in bcl-2/bax ratio prevented apoptosis in the presence of upregulated p53 and p21waf1/cip1. These findings indentified the involvement of multiple oncogenes in apoptosis regulation in vivo and demonstrate the complexity that may be associated with the use of a single oncogene assessment for predicting the outcome of cancer therapy with cytotoxic agents.
Apoptosis* ; Blotting, Western ; Cytotoxins ; Genes, Regulator ; Oncogenes ; Up-Regulation

BACKGROUND: It has been reported that immunological mechanisms may play an important role in the pathogenesis of vitiligo. Cellular degeneration in vitiligo is not limited to melanocytes but includes keratinocytes and probably whole epidermal melanin units. OBJECTIVE: The purpose of this study was to demonstrate the presence of anti-keratinocyte antibodies and the relationship between the level of antibodies and activity of the disease. METHODS: We analyzed the level of anti-keratinocyte antibodies in 13 patients with non-active, segmental vitiligo and 7 patients with active, non-segmental vitiligo by Western blotting. RESULTS: Using the Western blot method, we demonstrated the occurrence of anti-keratinocyte antibodies with keratinocyte cytoplasm. Anti-keratinocyte antibodies were more reactive in patients with active, non-segmental vitiligo than in those with non-active, segmental vitiligo. CONCLUSION: These results may be a secondary phenomenon to keratinocyte damage with concurrent occurrence of melanocyte destruction. However, the level of these antibodies may represent the acitivity of vitiligo.
Antibodies* ; Blotting, Western* ; Cytoplasm ; Humans* ; Keratinocytes* ; Melanins ; Melanocytes ; Vitiligo*

OBJECTIVE: Survivin is an inhibitor of apoptosis protein(IAP), which inhibits apoptosis through a pathway distinct from the Bcl-2 family members. Overexpression of survivin and Bcl-2 have been commonly reported in human neoplasms. The authors investigate whether there is a synergistic effect on the anti-apoptosis rate of primary brain tumors "in situ" based on the co-expression of survivin and Bcl-2. METHODS: One hundred and two brain tumor patients who had been resected were included in this study. Survivin and Bcl-2 were detected by Western blotting analysis, while apoptosis was examined by DNA fragmentation analysis. An anti-apoptotic rate was assessed in these brain tumor samples based on the expression of survivin and Bcl-2 or co-expression of both. RESULTS: Survivin and Bcl-2 were expressed in 57(55.9%) and 53(52.0%) of 102 brain tumor samples studied respectively, and co-expressed in 31(30.4%). The percentage of astrocytic and meningeal tumors expressing survivin was significantly correlated with histological grades; however, Bcl-2 was not correlated (p=0.106). The anti-apoptotic rate in primary brain tumors with survivin, Bcl-2, and both was detected in 49(86.0%) of 57 samples, 42(79.9%) of 53 samples, and 27(87.1%) of 31 samples, respectively. Their difference in the frequency of anti-apoptosis was not significant. CONCLUSION: Survivin or Bcl-2 is involved in the anti-apoptosis. However, it suggests that co-expression of survivin and Bcl-2, together, have no synergistic effect on the anti-apoptotic properties of the primary brain tumors.
Apoptosis ; Blotting, Western ; Brain Neoplasms* ; DNA Fragmentation ; Humans ; Meningeal Neoplasms