Proceedings of the Korean Society of Applied Pharmacology
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2007.11a
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pp.79-92
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2007
Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.
Rice seed maturation and germination involve drastic changes in water and nutrient transport, in which tonoplast aquaporins may play an important role. In the present study, gene expression profiles of 10 tonoplast intrinsic proteins (TIP) from rice were investigated by RT-PCR during seed development and germination. OsTIP3;1 and OsTIP3;2 were specifically expressed in mature seeds. Their transcript level rapidly decreased after onset of seed germination and gene expression was induced by ABA treatment. In contrast, expression of OsTIP2;1 and OsTIP4;3 was not seed specific as transcripts were found in vegetative tissues as well. Their respective transcript levels decreased at an early stage of seed development, whereas they increased at a later stage of seed germination and elongation of embryonic roots and shoots. When seed germination was inhibited by various stress conditions and ABA, expression of OsTIP2;1 and OsTIP4;3 was completely suppressed. In contrast, the expression level of OsTIP2;2 rapidly increased after seed imbibition and the transcript level was maintained under conditions inhibiting seed germination. These results implicate that tissue specific and developmental transcriptional regulation of OsTIPs in rice seeds depends on their specific function. In addition, OsTIPs can be discriminated by different potential phosphorylation and methylation sites in their protein structures. OsTIP3;1 and OsTIP3;2 possess unique phosphorylation signatures at their N-terminal domain, loop B and loop E, respectively. OsTIP2;1 and OsTIP4;3 have a potential methylation site at their Nterminal domain. This suggests that activity of specific tonoplast aquaporins may be regulated by post-translational modification as well as by transcriptional control.
Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.
We describe here a case of malignant mixed osteogenic tumor of the mammary gland with alveolar carcinomatous appreance. A firm, 2 to 2.5cm (in diameter) mass under the 5th nipple, showing the structure of extraosseous osteogenic sarcoma, was removed from the left 5th mammary gland of 12-year-old female dog. When investigated under the microscope, the osteoid material undergoing mineralization was surrounded by numerous scattered osteoblasts and a few osteoclastic cells throughout the osteoid tumorous stroma. The osteoid lesions were continuous with hypercellular myoepithelial cells of a very immature character with several mitotic figures. In addition, there were also carcinomatous tubules and alveoli, with invading cells into peripheral stroma, surrounded by myoepithelial cells in the mammary gland. In these lesions, emanating cords of tumor cells appear to be continuous with the myoepithelial cell layer of a duct. The presence of all these cell types suggests the existence of a common malignant origin, the stem cell being differentiated into epithelial carcinomatous and mesenchymal sarcomatous chondral and osteogenic tissues.
Purpose: Human Na+/I- symporter (hNIS) is known to be expressed in many tissues other than thyroid gland. The breast cancer cells are one of them and the possibility of radioiodine therapy in treatment of the breast cancer may be suggested. We investigated the expression rate of hNIS and the relationship between the expression of hNIS and the finding of 99mTc-MIBI scintimammographv in the breast cancer Materials and Methods: Surgically proved 56 patients with breast cancer were the subjects of this study The expression of hNIS were evaluated by immunohistochemistry and the results were compared to the findings of 99m7c-MIBI scintimammography. Results: Overall expression rate of hNIS was 41.1% in 56 patients. According to the pathologic diagnosis, it was 42.9% in 49 patients with invasive ductal carcinoma and 28.6% in the 7 patients with ductal carcinoma in situ. The expression rate of hNIS in the 41 cases with a focal increased uptake at he breast lesion on 99m7c-MIBI sointimammogram was 31.7%. That in the 15 cases without any abnormal uptake on the scan was significantly higher(65.7%, p<0.05). Conclusion: The expression rate of hNIS in the patients with breast cancer was not so high. The rate was higher in the patients with no increased uptake at the breast lesion on 99m7c-MIBI scintimammography.
An, Byung Ho;Yeon, Joon Ho;Kim, Soo Young;Choi, Sung Wook
The Korean Journal of Nuclear Medicine Technology
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v.19
no.1
/
pp.3-11
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2015
Purpose It is hard to obtain high quality images of knee and T.M joint because of a lot of soft tissues in the knee and T.M joint area. Most conventional system for high resolution scintigraphy was used by 4 mm aperture pinhole collimator. Performance comparison of high-resolution pinhole SPECT for Micro deluxe phantom using conventional system. the aim of this study is to evaluate performance of each aperture according to the diameter size and the usefulness of 24-hour delayed bone scintigraphy. Materials and Methods In this study 6 mm, 8 mm diameter pinhole collimators were mounted on Siemens E.CAM systems. In order to evaluate performance evaluation of each aperture and Micro Deluxe phantom was used for performance comparison of conventional SPECT system, Projection data were obtained with 9 degree increment per 30 second. Transverse images were reconstructed using dedicated OSEM algorithm with recovery of detector blurring. $^{99m}Tc-HDP$ source was used for 24-hour delayed bone scintigraphy. Results The knee joint images obtained with 24-hour delay were improved more than those obtained with 3-hour delay in our study. The 6 mm and 8 mm pinhole collimators FWHM have improved by 28% SNR and Uniformity have improved by 35%, Contrast has improved by 7% in 24-hour delayed knee joint image. While in 24-hour delayed T.M joint image of the 6 mm and 8 mm pinhole collimators FWHM have decreased by 60% SNR has decreased by 20% and Uniformity has decreased by 25%, Contrast has decreased significantly. Conclusion Pinhole collimators with 6 mm and 8 mm diameter could offer a superior performance for 24-hour delayed bone scintigraphy. The use of 24-hour delayed image provides additional benefits for pinhole scintigraphy of knee joint. Therefore, we expect that it is useful for precise diagnosis of knee joint and it is applicable to others joint imaging.
Kim Sang-Soo;Kim Kyung-Hee;Park Hyo-Jin;Hur Eun-hye;Rhim Hyangshuk
Journal of Life Science
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v.15
no.6
s.73
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pp.961-967
/
2005
Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.
Journal of The Korean Society of Grassland and Forage Science
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v.32
no.1
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pp.39-48
/
2012
This study was conducted to investigate effects of different levels of seleniferous whole crop barley (WCB) supplementation on performance, and blood characteristics as physiological responses in growing pigs. A total of 20 cross-bred pigs ((Landrace ${\times}$ Yorkshire) ${\times}$ Duroc) were divided into 4 treatments of 5 pigs each and experimental period lasted for 6 weeks. They were fed diets containing 0.1 (non-seleniferous WCB as controls), 0.2, 0.4, and 0.6 mg/kg levels of selenium (Se) by supplementing seleniferous WCB, and non-seleniferous or seleniferous WCB was formulated to 5% level in total ration. The diets were isonitrogenous (18% crude protein) and isocaloric (3,500 kcal/kg digestible energy) across treatments. Increasing levels of seleniferous WCB supplements did not affect feed intake and BW gain, and blood total protein concentration was (p<0.05) significantly higher for 0.2 mg/kg Se treatments than for controls. On d 14, blood albumin concentration was higher (p<0.05) for seleniferous WCB supplemented groups than for control group. Contrarily, blood glucose concentration was tended to be higher for controls than for seleniferous WCB groups. Blood total lipid concentration was significantly (p<0.05) lowered with increasing levels of seleniferous WCB. Serum glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase did not have any difference among treatments. It was tended that blood total cholesterol and triglyceride were lowered with increasing levels of seleniferous WCB. Blood Se concentration was significantly (p<0.05) increased with increasing levels of seleniferous WCB. The results indicate that Se present in seleniferous WCB had favorable effects on blood characteristics and blood Se increased by supplementing seleniferous WCB implies not only a good intestinal absorption of Se present in WCB but also the possibility of Se transfer into tissues.
In this study, we evaluated the effect of Capsosiphon fulvescens extract (CFE) and its active compound, pheophorbide A (PhA), on diabetic kidney failure. Diabetes mellitus (DM) was induced by a single intraperitoneal injection of streptozotocin (STZ; 40 mg/kg body weight (BW)). After a week, the rats were orally administered CFE (4 and 20 mg/kg BW) or PhA (0.2 mg/kg BW) once a day for 9 weeks. After scarification, renal tissue samples were collected for biochemical and histochemical analyses. Our study showed that the treatment with CFE and PhA significantly decreased lipid peroxidation level and the activities of glutathione peroxidase and glutathione-S-transferase (p<0.05), but it increased glutathione level and the activities of glutathione reductase, superoxide dismutase, and catalase in the renal tissues (p<0.05). The CFE- and PhA-treated rats with DM showed improved histochemical appearance and decreased abnormal glycogen accumulation. Therefore, we suggest that PhA-containing CFE could exert renal protective effects against STZ-induced oxidative stress.
This study was carried out to investigate the effects of uniconazole treatment on the growth and flowering of potted Chrysanthemum indicum L. for high quality pot plant production. Uniconazole was drenched at 0.05, 0.01, or 0.15 mg a.i./pot at 14 days after planting (DAP) of rooted cuttings. Simultaneously the short-day treatment (SDT) and pinching were adapted. The same amount of uniconazole (0.05 mg a.i./pot) was spilt drenched at once, twice, and three times, respectively, at 1 week interval. Uniconazole markedly reduced plant height, branch length, and stem diameter. Plant height was reduced linearly with increasing uniconazole concentration at 0.05, 0.01, or 0.15 mg a.i./pot up-to 41.6%, 52.5%, and 58.5%, respectively. In 0.05 mg a.i./pot, the number of branches greatly increased and plant height of 22.6 cm was adequate for pot plant. However, higher concentrations (0.10, 0.15 mg a.i.) were not suitable for production of high quality pot plant (17.0, 14.8 cm, respectively). Pinching and SDT decreased the number of days to visible bud, while uniconazole treatments delayed days to visible bud by 5-9 days compared with pinching and SDT. Number of visible buds was highest at 0.05 mg a.i./pot uniconazole treatment. However, flower diameter was decreased by uniconazole treatment, resulting in compact form. Number of stomata was increased by uniconazole treatment. The length of vascular tissues of uniconazole-treated plants ($11.2{\mu}m$) was smaller than that of non-treated plants ($15.0{\mu}m$, and the size of xylem vessel was also decreased. Uniconazole treatment at 0.05 mg a.i./pot at 14 DAP with pinching and SDT were recommended for pot plant production of C. indicum L.
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