• 한국수정란이식학회지
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• 제33권3호
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• pp.107-117
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• 2018

The purpose of this study is to expression pattern of melanogenesis associate genes on cultured melanocyte layer cells in Korean Brindle Cattle(Dark, Brindle and Yellow) were analyzed to evaluate the effects of sex hormones on the control of melanogenesis pathways. Korean Brindle Cattle(Dark, Brindle and Yellow) melanocyte in the skin cells was collected. after the addition of estrogen and testosterone, the culture was analyzed for expression of cell activity and melanin genes for 72 hours. For the analysis of estrogen in different coat color other than the melanogenesis-related genes it is increasingly yellow showed low expression. in particular, the cells of the brindle coat color is low active and expression of genes. However, the testosterone was low, the expression of cell activity inhibiting MMP-2. the expression of melanin genes actually showed a tendency to increase gradually, which is testosterone compared with the estrogen to be considered that affect the skin cell layer brindle coat color. In this study, stimulation with estrogen triggered the inhibition of MC1R of the melanocyte in brindle coat color, but testosterone is induced MC1R in melanocyte. Therefore, considered the eumelanin or phaeomelanin activation are controlled caused by differential expression of sex hormones on melanocyte in Korean Brindle Cattle.

• The Plant Pathology Journal
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• 제20권4호
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• pp.258-263
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• 2004

Ripe fruit of pepper (Capsicum annuum) showed resistance to Colletotrichum gloeoporioides, but unripe fruit was susceptible. We previously isolated the PepTLP gene that induced in both unripe and ripe fruit by fungal infection and wound, and only in ripe fruit by jasmonic acid (JA) treatment. To examine further regulation of PepTLP, the action of specific agonist and antagonists of known signaling effector on the .PepTLP expression by fungal infection, wound, and JA was investigated. A similar dephosphorylation event negatively activated all the PepTLP expression in the ripe fruit by fungal infection, wound, and JA. The induction of PepTLP expression by wound is differentially regulated via phosphorylation and dephosphorylation step during pre- and post-ripening, respectively. In addition, the induction of PepTLP expression in the ripe fruit by wound and JA is differentially regulated via dephosphorylation and phosphorylation step, respectively. Only both wound and JA treatment has synergistic effect on the PepTLP expression in the unripe fruit. Both SA and JA treatments on the unripe fruit, and both wound or JA and SA on the ripe fruit could not do any effect on the expression of PepTLP. These results suggest that the induction of PepTLP expression is differentially regulated via complex regulatory system against fungal infection, wound, and JA treatment during pre- and post-ripening of pepper fruit.

• The Plant Pathology Journal
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• 제36권5호
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• pp.450-458
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• 2020

Pepper and tomato plants infected with two Clavibacter species, C. capsici and C. michiganensis have shown different patterns of disease development depending on their virulence. Here, we investigated how pepper and tomato plants respond to infection by the high-virulent or low-virulent Clavibacter strains. For this, we chose two strains of each Clavibacter species to show different virulence level in the host plants. Although low-virulent strains showed less disease symptoms, they grew almost the same level as the high-virulent strains in both plants. To further examine the response of host plants to Clavibacter infection, we analyzed the expression patterns of plant defense-related genes in the leaves inoculated with different strains of C. capsici and C. michiganensis. Pepper plants infected with high-virulent C. capsici strain highly induced the expression of CaPR1, CaDEF, CaPR4b, CaPR10, and CaLOX1 at 5 days after inoculation (dai), but their expression was much less in low-virulent Clavibacter infection. Expression of CaSAR8.2 was induced at 2 dai, regardless of virulence level. Expression of GluA, Pin2, and PR2 in tomato plants infected with high-virulent C. michiganensis were much higher at 5 dai, compared with mock or low-virulent strain. Expression of PR1a, Osmotin-like, Chitinase, and Chitinase class 2 was increased, regardless of virulence level. Expression of LoxA gene was not affected by Clavibacter inoculation. These results suggested that Clavibacter infection promotes induction of certain defense-related genes in host plants and that differential expression of those genes by low-virulent Clavibacter infection might be affected by their endophytic lifestyle in plants.

• Fisheries and Aquatic Sciences
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• 제18권1호
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• pp.73-80
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• 2015

To examine the interrelationship between transgenic insertion patterns and transgene expression profiles in established transgenic fish lines, four stable transgenic marine medaka Oryzias dancena germlines harboring ${\beta}$-actin regulator-driven RFP reporter constructs were selected. The established transgenic strains were characterized with regard to their transgenic genotypes (insertion pattern, concatemer formation, and transgene copy number based on genomic Southern blot hybridization and qPCR assay) and expression characteristics at the mRNA (qRT-PCR), protein (western blot), and phenotypic (fluorescent appearance) levels. From comparative examinations, it was found that transgenic expression at both the transcription and translation levels could be significantly downregulated in transgenic strains, potentially through methylation-mediated transgene silencing that was particularly associated with the formation of a long tail-to-head tandem concatemer in the chromosomal integration site(s). When this occurred, an inverse relationship between the transgene copy number and fluorescence intensity was observed in the resultant transgenic fish. However, with the other transgenic genotype, transgenic individuals with an identical Southern blot hybridization pattern, containing a tandem concatemer(s), had very different expression levels (highly robust vs. low expression strengths), which was possibly related to the differential epigenetic modifications and/or degrees of methylation. The concatemer-dependent downregulation of transgene activity could be induced in transgenic fish, but the overall pattern was strain-specific. Our data suggest that neither a low (or single) transgene copy number nor tandem transgene concatemerization is indicative of strong or silenced transgene expression in transgenic fish carrying a ubiquitous transgene. Hence, a sufficient number of transgenic lineages, with different genotypes, should be considered to ensure the establishment of the best-performance transgenic line(s) for practical applications.

• Toxicological Research
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• 제20권3호
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• pp.233-239
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• 2004

The present study was designed to determine the impact of mercury on endotoxin-induced inflammatory cytokine expression and corresponding signal transduction in mouse kidney. Male BALB/c mice were exposed continuously to 0, 0.3, 1.5, 7.5, or 37.5 ppm of mercury in drink-ing water for 14 days and at the end of the treatment period, lipopolysaccharide (LPS, 0.5 mg/kg) was injected intraperitoneally 2 h prior to euthanasia. The doses of mercury and LPS did not cause hepatotoxicity or renal toxicity as indicated by unaltered plasma alanine aminotransferase and aspartate aminotransferase levels, and terminal UTP nucleotide end-labeling assay from kidney, respectively. Mercury decreased kidney glutathione (GSH) and with LPS, it additively decreased GSH. Mercury activated p38 mitogen-activated protein kinase (MAPK) and additively increased LPS-induced p38 MAPK phosphorylation. In contrast, mercury inhibited LPS-induced activation of extra-cellular signal-regulated kinase (ERK) but had no effect alone. Mercury increased the gene expression of tumor necrosis factor $\alpha$ (TN F$\alpha$) and potentiated LPS-induced TNF$\alpha$ expression. Mercury did not affect LPS-induced interleukin-1$\beta$ (IL-1$\beta$) expression but decreased LPS-induced IL-6 expression. These results suggest that low levels of mercury might augment LPS-induced TNF$\alpha$ expression by altering GSH and p38 MAPK. Mercury modulates LPS-induced p38 and ERK activation, and downstream TNF$\alpha$ and IL-6 expression in kidney, respectively.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7201-7205
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• 2014

Chemokine and chemokine receptor expression by tumor cells contributes to tumor growth and angiogenesis and thus these factors may be considered as tumor markers. Here we aimed to characterize cells directly extracted from glioma, meningioma, and secondary brain tumors as well as non-tumoral cells in vitro. Cells were isolated from brain tissues using 0.2% collagenase and characterized by flow cytometry. Expression of SDF-1, CXCR4, CXCR7, RANTES, CCR5, MCP-1 and IP-10 was defined using flow cytometry and qRT-PCR methods. Brain tissue isolated cells were observed as spindle-shaped cell populations. No significant differences were observed for expression of SDF-1, CXCR4, CXCR7, RANTES, CCR5, and IP-10 transcripts. However, the expression of CXCR4 was approximately 13-fold and 110-fold higher than its counterpart, CXCR7, in meningioma and glioma cells, respectively. CXCR7 was not detectable in secondary tumors but CXCR4 was expressed. In non tumoral cells, CXCR7 had 1.3-fold higher mRNA expression than CXCR4. Flow cytometry analyses of RANTES, MCP-1, IP-10, CCR5 and CXCR4 expression showed no significant difference between low and high grade gliomas. Differential expression of CXCR4 and CXCR7 in brain tumors derived cells compared to non-tumoral samples may have crucial impacts on therapeutic interventions targeting the SDF-1/CXCR4/CXCR7 axis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제38권2호
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• pp.78-84
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• 2012

Objectives: Odontogenic keratocysts (OKCs) have a tendency to recur and possess an aggressive nature. the aim of the present study was to evaluate cytokeratin (CK) expression patterns as a method for the differentiation between dentigerous cysts (DCs) and OKCs, as their histomorphologic appearance are often indistinguishable. Materials and Methods: Formalin-fixed, paraffin-embedded tissue sections of 43 OKCs and 38 DCs were immunohistochemically analyzed with i-solution in a quantitative manner in order to evaluate the immunoreactivity of CK 10, 16 and 17. Results: CK 10 expression was evident in 79.1% of OKCs but found in only 18.4% of DCs (P<0.05), and CK 10 expression was observed to occur more frequently in OKCs (mean 25.45%) than in DCs (2.19%) (P<0.05). The expression of CK 16 was evident in 79.1% of OKCs but found in only 7.9% of the DCs (P<0.05) and CK 16 expression was observed to occur more frequently in OKCs (mean 4.33%) than in the DCs (0.61%) (P<0.05). The expression of CK 17 was evident in 88.4% of OKCs but seen in only 15.7% of the DCs (P<0.05) and CK 17 expression was observed to occur more frequently in OKCs (mean 31.11%) than in the DCs (2.37%) (P<0.05). Conclusion: The immunohistochemical detection of CK 10, 16 and 17 can be utilized as a valuable biomarker for use in distinguishing between OKCs and DCs, which have clinically significant differential diagnoses.

• Archives of Pharmacal Research
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• 제26권12호
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• pp.1079-1086
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• 2003

The aim of this study was to investigate the effect of trauma on cytochrome P450 (CYP) gene expression and to determine the role of Kupffer cells in trauma-induced alteration of CYP isozymes. Rats underwent closed femur fracture (FFx) with associated soft-tissue injury under anesthesia. To deplete Kupffer cells in vivo, gadolinium chloride ($GdCl_3$) was intravenously injected at 7.5 mg/kg body wt., 1 and 2 days prior to FFx surgery. At 72 h of FFx, liver tissues were isolated to determine the mRNA and protein expression of CYP isozymes and NADPH-P450 reductase by reverse transcription-polymerase chain reaction and Western immunoblotting, respectively. In addition, the mRNA levels of tumor necrosis factor alpha (TNF-$\alpha$), inducible nitric oxide synthase (iNOS) and heme oxygenase-1 (HO-1) were evaluated. FFx increased the mRNA level of CYP1A1; an increase that was not prevented by $GdCl_3$. There were no significant differences in the mRNA expression of CYP1A2, 2B1 and 2E1 among any of the experimental groups. The protein levels of CYP2B1 and 2E1 were significantly decreased by FFx; a decrease that was not prevented by $GdCl_3$ treatment. The gene expression of NADPH-P450 reductase was unchanged by FFx. FFx significantly increased the expression of TNF-$\alpha$ mRNA; an increase that was attenuated by $GdCl_3$. The mRNA expression of HO-1 was increased by FFx, but not by $GdCl_3$ . Our findings suggest that FFx differentially regulates the expression of CYP isozyme through Kupffer cell-independent mechanisms.

• 대한의생명과학회지
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• 제16권2호
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• pp.105-112
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• 2010

Methylprednisolone (MPD) is a synthetic glucocorticoid drug used in treatment of many neurological diseases and neurotraumas, including spinal cord injuries. Little is known of the mechanism of MPD in neuronal cells, particularly the genetic expression aspect. DD-PCR was used in identification of genes expressed during MPD treatment of PC12 cells. We have isolated 3 predicted up- or down-regulated genes, which are differentially expressed in neurons by MPD. One of these genes, USP16 (ubiquitin specific protease 16), is the deubiquitinating enzyme that is up-regulated by MPD in neurons. In order to observe the effect of MPD on USP16 gene expression, PC12 cells were treated under several experimental conditions, including endoplasmic reticulum stress drugs. We have isolated the total RNAs in PC12 cells and detected USP16 and ER related genes by RT-PCR. Because its expression pattern is similar to expression of ER chaperons, USP16 gene expression is strongly associated with unfolded protein response. A meaningful negative effect on each tissue treated by methylprednisolone is not shown in vivo. USP16 gene expression is suppressed by LY294002 (phosphatidylinositol 3-kinase inhibitor), which suggests that USP16 gene expression is regulated by the phosphatidylinositol 3-kinase pathway.

• 대한두경부종양학회지
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• 제21권1호
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• pp.15-20
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• 2005

Objectives: The hepatocyte growth factor(HGF)/c-Met pathway may play various roles in the carcinogenesis of various organs. Although HGF/c-Met signalling pathway has been shown to demonstrate various cellular responses including mitogenic, proliferative, morphogenic and angiogenic activities, the study on their expression related to clinicopathological parameters in thyroid tumor is relatively rare. So we want to find out the clinical significance of the c-Met in thyroid tumor. Materials and Methods: We assess the mRNA and protein expression of the c-Met genes by means of RT-PCR method and the immunohistochemical stain in 100 cases of thyroid tumors(50 papillary carcinomas, 10 follicular carcinomas, 20 follicular adenomas, 20 nodular hyperplasia). Results: By RT-PCR, c-Met mRNA was detected in 43(86%) in papillary carcinoma, 4(40%) in follicular carcinoma, 4(20%) in follicular adenoma and 2(10%) in nodular hyperplasia cases. By immunohistochemistry, c-Met protein expression was detected in 44(88%), 2(20%), 3(15%) and 1(5%). Expression of the c-Met mRNA and protein expression was significantly highly recognized in papillary carcinoma. The c-Met protein overexpression was significantly correlated with the grade of the differentiation. Conclusion: These results suggest that c-Met expression may be associated with thyroid papillary cancer progression. The differential expression of c-Met protein and mRNA suggests that these molecules may be a reliable diagnostic marker in thyroid papillary cancer.