Ham, Young-An;Choi, Hyun-Jin;Kim, Soo-Hyun;Chung, Mi-Ja;Ham, Seung-Shi
Journal of the Korean Society of Food Science and Nutrition
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v.38
no.1
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pp.25-31
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2009
This study was carried out to investigate the mutagenic, antimutagenic, cytotoxicity and antitumor effects of Adenophora triphylla (AT). AT was extracted with 70% ethanol and then further fractionated to hexane, chloroform, ethyl acetate, butanol and water. Antimutagenic, cytotoxicity and antitumor effects of AT extracts were measured by using Ames test, SRB method, and the tumor growth inhibition test. AT extracts did not show any mutagenicity in the Ames test; however, 70% ethanol extracts and its fractions had strong antimutagenic effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The ethyl acetate fraction of AT (200 ${\mu}g$/plate) showed approximately 66.5% inhibitory effect on the mutagenesis induced by 4NQO against TA98 strain, whereas 83.3% and 75.1% inhibitions were observed on the mutagenesis induced by MNNG and 4NQO against TA100 strain. In anticancer effects, the cytotoxicity of AT extract and its fractions against cancer cell lines including human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human breast adenocarcinoma (MCF-7), human gastric carcinoma (AGS), human lung carcinoma (A549) and transformed primary human embryo kidney (293) were investigated. The treatment of 1 mg/mL AT ethyl acetate faction had the highest cytotoxicity of 79.9%, 74.9%, 66.0%, 71.0% and 74.3% against HeLa, Hep3B, MCF-7, AGS and A549 cells, respectively. In contrast, the extract and its fractions showed only $3{\sim}36%$ cytotoxicity for a normal human kidney cell line (293). In vivo anti-cancer effect of Adenophora triphylla extract was tested using Balb/c mice transplanted sarcoma-180 cells. Adenophora triphylla ethyl acetate fraction showed the highest inhibition rate of 37.2% at the 50 mg/kg concentration.
Background: Fascin is an actin-bundling protein that induces membrane protrusions and it increases cell motility in various transformed cells. Esophageal cancer is one of the most lethal malignancies, and it exhibits extensive local invasion or frequent regional lymph node metastasis even after curative surgery. We investigate the expression of fascin by performing immunohistochemistry to evaluate the clinical characteristics and prognostic significance of its expression in esophageal cancer patients. Material and Method: Immunochemistry for fascin was performed on 76 tumor samples from 76 patients who underwent esophageal cancer operations. The expression levels of fascin in the 76 esophageal cancer tissues were compared with those in the corresponding normal esophageal epithelium. The fascin-positive samples were defined as those showing more than 75% of fascin-positive cells. Result: Overall, a fascin positive expression was detected in 39 (51.3%) out of the total 76 cases. The tumors with positive fascin expression tended to more frequently show a higher stage (p=0.030), and a higher T-factor (p=0.031). The prognosis of the fascin negative group was significantly better than that of the fascin positive group (p=0.004). Multivariate analysis revealed that lymphovascular invasion and the fascin expression were independent prognostic factors. Conclusion: Fascin was expressed in 513% of the esophageal cancer tissues and a positive expression of fascin was associated with more advanced tumor progression and recurrence. Our study suggests that the fascin expression may be an independent prognostic factor for an unfavorable clinical course few those patients suffering with esophageal cancer.
It has been known that ras signaling transduction leads to cell proliferation and migration including various adaptor molecules. Dynamin protein has been implicated in the formation of nascent vesicles in both the endocytic and secretory pathways. Dynamin was classified into three isoforms: dynamin I is only expressed in neuronal tissue, dynamin II is expressed ubiquitously in all tissue but that of dynamin III is confined to testis. We have reported in previous study that Grb2, binding to ras, was associated with dynamin II in NIH3T3 cells. Therefore we have tried to identify the relative expression of dynamin II according to overexpressed ras protein in ras oncogene transfected cells (NIH3T3 (ras)). For the detection of differential expression of dynamin II, we have used immunofluorescent staining and western blot methods in NIH3T3 and NIH3T3 (ras) cells. Next we have described the morphological differences between NIH3T3 and NIH3T3 (ras) cells using SEM and TEM. From these experiments dynamin II was highly expressed in NIH3T3 (ras) cells. NIH3T3 cells was transformed to more spindle shape with many cell process by transfection of ras oncogene. Moreover dynamin II was more concentrated in endocytotic membrane of the NIH3T3 (ras) cells compared to that of NIH3T3 cells. The present results suggested that dynamin II may involve the intermediate messenger in Ras signaling transduction pathway.
Responsiveness of muscarinic and alpha adrenoceptor activation on endothelial cells was studied in isolated canine renal artery rings. Ach (10-100 nM), dose dependently, relaxes endothelial intact rings precontracted with phenylephrine ($IC_{50}$ of Ach was 34.5 nM). Selective mechanical destruction of the endothelium transformed the activity of this substance from vasodilatation to vasoconstriction. Acetylcholine induced relaxations could be selectively inhibited competitively by atropine, but could not be inhibited by cyclooxygenase inhibitor. Methylene blue, however, an inhibitor of soluble guanylate cyclase activity, inhibited Ach as well as sodium nitroprusside (SNP) induced relaxation. Relaxation produced by prostacyclin was not modified by methylene blue. On the other hand, alpha adrenoceptor agonist did not relax but contract canine renal artery rings possessing an intact intima precontracted with U-46619. Clonidine, however, selective alpha-2 adrenergic agonist, is more susceptible than phenylepherine, selective alpha-1 adrenergic agonist, to the inhibitory effect of contraction. These results suggest that in canine renal artery rings, 1) muscarinic receptor is responsible for releasing endothelium dependent relaxation factor (EDRF). 2) alpha-1 and alpha-2 adrenergic receptors are present in canine renal artery. 3) relaxation via EDRF is antagonized by methylene blue, providing further evidence that EDRF acts through a cGMP mechanism.
Today using Internet environment is considered absolutely essential for establishing corporate marketing strategy. Companies have promoted their products and services through various ways of on-line marketing activities such as providing gifts and points to customers in exchange for participating in events, which is based on customers' membership data. Since companies can use these membership data to enhance their marketing efforts through various data analysis, appropriate website membership management may play an important role in increasing the effectiveness of on-line marketing campaign. Despite the growing interests in proper membership management, however, there have been difficulties in identifying inappropriate members who can weaken on-line marketing effectiveness. In on-line environment, customers tend to not reveal themselves clearly compared to off-line market. Customers who have malicious intent are able to create duplicate IDs by using others' names illegally or faking login information during joining membership. Since the duplicate members are likely to intercept gifts and points that should be sent to appropriate customers who deserve them, this can result in ineffective marketing efforts. Considering that the number of website members and its related marketing costs are significantly increasing, it is necessary for companies to find efficient ways to screen and exclude unfavorable troublemakers who are duplicate members. With this motivation, this study proposes an approach for managing duplicate membership based on the social network analysis and verifies its effectiveness using membership data gathered from real websites. A social network is a social structure made up of actors called nodes, which are tied by one or more specific types of interdependency. Social networks represent the relationship between the nodes and show the direction and strength of the relationship. Various analytical techniques have been proposed based on the social relationships, such as centrality analysis, structural holes analysis, structural equivalents analysis, and so on. Component analysis, one of the social network analysis techniques, deals with the sub-networks that form meaningful information in the group connection. We propose a method for managing duplicate memberships using component analysis. The procedure is as follows. First step is to identify membership attributes that will be used for analyzing relationship patterns among memberships. Membership attributes include ID, telephone number, address, posting time, IP address, and so on. Second step is to compose social matrices based on the identified membership attributes and aggregate the values of each social matrix into a combined social matrix. The combined social matrix represents how strong pairs of nodes are connected together. When a pair of nodes is strongly connected, we expect that those nodes are likely to be duplicate memberships. The combined social matrix is transformed into a binary matrix with '0' or '1' of cell values using a relationship criterion that determines whether the membership is duplicate or not. Third step is to conduct a component analysis for the combined social matrix in order to identify component nodes and isolated nodes. Fourth, identify the number of real memberships and calculate the reliability of website membership based on the component analysis results. The proposed procedure was applied to three real websites operated by a pharmaceutical company. The empirical results showed that the proposed method was superior to the traditional database approach using simple address comparison. In conclusion, this study is expected to shed some light on how social network analysis can enhance a reliable on-line marketing performance by efficiently and effectively identifying duplicate memberships of websites.
For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.
Black rockfish Sebastes schlegeli is one of scopaenid fish giving birth to yolk-sac larvae and distributed around the coast of Korea and Japan, and is one of main species which is cultured in Korea. The difffrentiation of thyroid gland and the changes of thyroid hormones (THs) concentrations in the whole body during early development of this species were examined and the relationship between thyroid gland and their growth will be presented. Total length (TL) and body weight (BW) of the larva at the parturition were 6.3${\pm}$0.1 mm and 3.6${\pm}$0.1 mg, respectively. The larvae transformed to juvenile after 30th day after parturition. According to Hoshiai(1977), they had gown into stage Fl (TL 13.2${\pm}$0.9 mm, BW 46.5${\pm}$1.5 mg) at 50th day. The thyroid gland of black rockfish was first observed histologically in hatching larvae in mother fish. The larvae just after parturition have 1${\sim}$3 of the thyroid follicles diffrentiated between basibranchial bone and ventral aorta, at the base of the first gill arch. In this time, thyroid follic1e number (TFN),thyroid follicle diameter (TFD) and thyroid cell height (TCH) were 1.6${\pm}$0.8 pieces/inds., 18.1${\pm}$0.6 ${\mu}$m and 4.1${\pm}$0.2 ${\mu}$m, respectively. TFN and TFD at 50th day were increased to 32.5${\pm}$6.9 pieces/inds. and 41.5${\pm}$1.7 ${\mu}$m, respectively. These results indicate that they are related to the growth of black rockfish during early development. However, TCH indicates that the activity of thyroid gland appearedat special day, eg. 5, 20 and 50th day, suggesting that TCH may have some role in the physiological activity. L-thyroxine (T$_4$)concentration decreased sharply to 10 days after parturition, and at 25th day (metamorphosis stage) increased markedly to 3.44${\pm}$0.93ng/g fish. After this time, T$_4$ concentration decreased at 35th day, but then increasedagain to the highest concentration, 5.63${\pm}$0.70ng/g fish. 3,5,3'-triiodo-L-thyronine (T$_3$) concentration declined sharply from just after pafurition (4.96${\pm}$1.90 ng/g fish) to 5th day (0.30${\pm}$0.07 ng/g fish). However T$_3$ concentration increased markedly to 0.95${\pm}$0.11 ng/g fish at 30th day and then did not significantly change until 45th day, increased also sharply to 1.67${\pm}$0.23 ng/g fish at 50th day.
Ethanol is a very valuable material, however, it is also a source of stress, as the accumulation of ethanol in a medium inhibits cell viability and decreases productivity of the target product. Therefore, the ethanol tolerance of yeast, which is closely related to ethanol productivity, is an important factor in industrial ethanol production. In this study, the YDJ1 and PEP5 genes were selected as target genes for elucidating ethanol-tolerant mechanisms by analyzing the expression regulation of these genes. The pA-YDJ1 and pA-PEP5 plasmids containing YDJ1 and PEP5 genes under an ADH1 promoter, respectively, were constructed and transformed into BY4742 (host strain), BY4742△ydj1, and BY4742△pep5 strains. The ethanol tolerance in the BY4742△ydj1/ pA-YDJ1 and BY4742△pep5/pA-PEP5 transformants was restored by overexpression of the YDJ1 and PEP5 genes to the host strain level. The YDJ1 and PEP5 genes were also introduced into the double gene disruptant (BY4742△ydj1△pep5) to investigate the expression regulation of the YDJ1 and PEP5 genes. The simultaneous overexpression of the YDJ1 and PEP5 genes restored ethanol tolerance to the 90% level of the BY4742 strain under 8% ethanol stress. The YDJ1 gene induced more overexpression of the PEP5 gene in the BY4742△ydj1 △pep5/pA-YDJ1, pA-PEP5 strain, suggesting that the YDJ1 gene partially regulates the expression of the PEP5 gene as an upstream regulator.
Surface data generally represent continuous distribution of geographical or social phenomena of a region in urban analysis. Instances include distribution of temperature, population of region, and various distributions related to human activities. When spatial data are given in the form of surface, surface comparison is required as a way of comprehending the surface change or the relationship between two surfaces. As for previous approaches of surface comparison, there are visualization, quantitative methods and qualitative method. All those approaches, however, show the difference between two surfaces in a limited way. Especially, they are not able to distinguish spatial difference between two surfaces. To overcome such problem, this paper proposes a method of comparing two surfaces in terms of their spatial structure. Main concept of the method comes from earth moving problem and the method is named minimum surface transformation, here. When a surface is transformed into another, total surface volume moved in the process of transformation should be the minimum. Both quantitative and spatial differences between two surfaces are evaluted by total surface volume moved and the distribution of moved surface volume of each cell respectively. The method is applied to hypothetical and actual data. From the former, it is understood that the method explains how two surfaces are quantitatively and spatially different. The result of the latter shows that moved total surface volume decreases as time goes by which fits the actual situation that population change rate gets smaller. Concerning the other measure of surface difference, the distribution of $X_{ij}$ describes detailed flow of surface volume than that of simply subtracting surface volume by indicating to what direction the population change occurs.
Kim, Young-Kyoon;Kim, Seung-Joon;Park, Yong-Keun;Kim, Seok-Chan;Kim, Kwan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak;Kim, Sang-Ho
Tuberculosis and Respiratory Diseases
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v.49
no.6
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pp.691-702
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2000
Background : Acute lung injury (ALI) is a commonly encountered respiratory disease and its prognosis is poor when the treatment is not provided promptly and properly. However no specific pharmacologic treatment is currently available for ALI, although recently several supportive drugs have been under scrutiny. We studied anti-inflammatory effects of pentoxifylline (PF), a methylated xanthine, and ONO-5046, a synthetic neutrophil elastase inhibitor on lipopolysaccharide (LPS)-induced ALI in vitro. Methods : To establish an in vitro model of LPS-induced ALI, primary rat alveolar macrophages and peripheral neutrophils in various ratios (1:0, 5:1, 1:1, 1:5, 0:1) were co-cultured with transformed rat alveolar epithelial cells (L2 cell line) or vascular endothelial cells (IP2-E4 cell line) under LPS stimulation. Each experiment was divided into five groups-control, LPS, LPS+PF, LPS+ONO, and LPS+PF+ONO. We compared LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils in various ratios, and the resultant cytotoxicity on L2 cells or IP2-E4 cells between groups. In addition we also compared the productions of tumor necrosis factor (TNF)-$\alpha$ interleukin (IL)-$1{\beta}$, monocyte chemotactic protein(MCP)-1, IL-6, and IL-10 as well as mRNA expressions of TNF-$\alpha$ inducible nitric oxide synthetase(iNOS), and MCP-1 from LPS-stimulated primary rat alveolar macrophages between groups. Results : (1) PF and ONO-5046 in each or both showed a trend to suppress LPS-induced superoxide anion productions from primary rat alveolar macrophages and peripheral neutrophils regardless of their ratio, except for the LPS+PF+ONO group with the 1:5 ratio, although statistical significance was limited to a few selected experimental conditions. (2) PF and ONO-5046 in each or both showed a trend to prevent IP2-E4 cells from LPS-induced cytotoxicity by primary rat alveolar macrophages and peripheral neutrophils regardless their ratio, although statistical significance was limited to a few selected experimental conditions. the effects of PF and/or ONO-5046 on LPS-induced L2 cell cytotoxicity varied according to experimental conditions. (3) PF showed a trend to inhibit LPS-induced productions of INF-$\alpha$ MCP-1, and IL-10 from primary rat alveolar macrophages. ONO-5046 alone didnot affect the LPS-induced productions of proinflammatory cytokines from primary rat alveolar macrophages but the combination of PF and ONO-5046 showed a trend to suppress LPS-induced productions of INF-$\alpha$ and IL-10 PF and ONO-5046 in each or both showed a trend to increase LPS-induced IL-$\beta$ and IL-6 productions from primary rat alveolar macrophages. (4) PF and ONO-5046 in each or both showed a trend to attenuate LPS-induced mRNA expressions of TNF-$\alpha$ and MCP-1 from primary rat alveolar macrophages but at the same time showed a trend increase iNOS mRNA expression. Conclusion : These results suggest that PF and ONO-5046 may play a role in attenuating inflammation in LPS-induced ALI and that further study is needed to use these drugs as a new supportive therapeutic strategy for ALI.
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