• Title/Summary/Keyword: $\beta$-actin

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Immunomodulatory effects of chlorogenic acid and ethyl acetate fraction from Lonicera japonica on cytokine gene expression profiles in spleen and thymus (Chlorogenic acid 및 인동등 ethyl acetate 분획의 비장 및 흉선 세포에서의 유전자 발현 분석을 통한 면역조절효과)

  • Ha, Tae-Kwang;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.1-10
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    • 2011
  • Objective : Lonicera japonica contains anti complementary polysaccharides and polyphenolic compound. Among these polyphenolic substances, chlorogenic acid is the major active component of this plant. However, the immunological mechanisms for these activities, have not been elucidated, nor the active components. To clarify immunomodulatory effects of those we examined the relationship between the activity of CD8+ T cell-mediated lysis and the frequency of cytokine profiles in spleen, thymus (especially IFN-${\gamma}$, IL-4, GM-CSF etc.) expressing CD8+ T cells activated by IL-2. Methods : To study immunomodulatory effects ethyl acetate fraction from Lonicera japonica, chlorogenic acid on cytokine gene expression from spleen, thymus cells, RT-PCR was performed after quantitative normalization for each gene by a densitometry using ${\beta}$-actin gene expression. A modified standard $^{51}Cr$-release assay was used to measure cytotoxic activities of cytotoxic T cells. Spleen, thymus cells from NOD mice were stained with CD3, CD4, CD44, CD69 in staining buffer and analyzed by two color flow cytometry. Results : We showed that ethyl acetate fraction from Lonicera japonica in combination with IL-2 resulted in a significant enhancement of PCR products for IFN-${\gamma}$, IL-4, IL-10, GM-CSF, IL-6 and cytotoxtic CD8+ T cell proportion in spleen and thymus T cells in NOD mice. This suggests that IFN-${\gamma}$, IL-6 like IL-4 may be acting as a regulatory rather than proinflammatory cytokine. Conclusions : In conclusion, based on the results of the present study which showed that ethyl acetate fraction from Lonicera japonica and chlorogenic acid upregulating cytokine gene expression in spleen and thymus, we are tempted to speculate that some of the therapeutic efficacies such as anti-diabetic activity of Lonicera japonica are due to the immunomodulatory its ethylacetate fraction and chlorogenic acid.

Retrovirus Vector-Mediated Inductional Expression of the Human Lactadherin Gene in Mouse Mammary Epithelial Cells (Mouse Mammary Epithelial Cell에서 Retrovirus Vector를 이용한 Human Lactadherin 유전자의 유도적 발현)

  • 권모선;구본철;정병현;염행철;박창식;김태완
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.15-23
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    • 2003
  • Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of tissue specific and hormonal inducible mouse whey acidic protein (WAP) promote., the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAP promoter was accomplished in the presence of insulin, hydrocortisone and prolactin. Compared to the control (cells cultured with insulin alone), however we observed that the WAP promoter was leaky. These data indicate that luther studies are needed in finding an appropriate promoter other than WAP promoter because of its leakiness.

First Report of Colletotrichum fioriniae Causing Anthracnose on Fruit of Pumpkin (Cucurbita moschata) in Korea (Colletotrichum fioriniae에 의한 호박 과실 탄저병의 발생 보고)

  • Kim, Joon-Young;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.26 no.3
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    • pp.190-193
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    • 2020
  • In 2014, Anthracnose symptoms were observed on fruit of pumpkin in Gangneung, Gangwon in Korea. The typical symptoms on fruit surfaces were circular lesions, brown, shriveled with salmon-colored spore masses on the spots. A fungal isolate was obtained from symptomatic on fruit and cultured on potato dextrose agar at 25℃. The color of the upper surface of the colony varied from white to gray with occurrence of salmon-colored conidial masses and was red to violet on the underside. The causal fungus of anthracnose isolated from the diseased fruits was identified as Colletotrichum fioriniae based on mycological characteristics and sequences of the internal transcribed spacer, glyceraldehyde-3-phosphate dehydrogenase, chitin synthase-1 gene, histone H3, actin gene, beta-tubulin gene region. Pathogenicity tests of the isolate were performed on pumpkin fruit by both wound and non-wound inoculation with conidial suspension (1×106 conidia/ml) and incubated at 25℃. Lesions were observed on both wounded and non-wounded fruit of pumpkin after 7 days inoculation in only C. fioriniae. The pathogen was re-isolated from the lesions of inoculated fruits. To our knowledge, this is the first report of anthracnose on fruit of pumpkin caused by C. fioriniae in Korea.

Proteomic Analysis of Bovine Muscle Satellite Cells during Myogenic Differentiation

  • Rajesh, Ramanna Valmiki;Jang, Eun-Jeong;Choi, In-Ho;Heo, Kang-Nyeong;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1288-1302
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    • 2011
  • The aim of this study was to analyze the proteome expression of bovine satellite cells from longissimus dorsi (LD), deep pectoral (DP) and semitendinosus (ST) muscle depots during in vitro myogenic differentiation. Proteomic profiling by twodimensional gel electrophoresis and mass spectrometry of differentiating satellite cells revealed a total of 38 proteins that were differentially regulated among the three depots. Among differentially regulated proteins, metabolic proteins like lactate dehydrogenase (LDH), malate dehydrogenase (MDH) were found to be up regulated in ST, while alpha-enolase (NNE) in LD and DP depot satellite cells were down regulated. Also, our analysis found that there was a prominent up regulation of cytoskeletal proteins like actin, actincapping protein and transgelin along with chaperone proteins like heat shock protein beta 1 (HSPB 1) and T-complex protein 1 (TCP-1). Among other up regulated proteins, LIM domain containing protein, annexin 2 and Rho GDP-dissociation inhibitor 1 (Rho GDI) are observed, which were already proven to be involved in the myogeneis. More interestingly, satellite cells from ST depot were found to have a higher myotube formation rate than the cells from the other two depots. Taken together, our results demonstrated that, proteins involved in glucose metabolism, cytoskeletal modeling and protein folding plays a key role in the myogenic differentiation of bovine satellite cells.

Toxic Effects of Fungicide Tebuconazole on the Early Development of African Clawed Frog, Xenopus laevis (진균제 농약 tebuconazole이 Xenopus laevis의 초기 배 발생에 미치는 독성 영향)

  • Hwang, Yong-Gi;Lee, Mi-Ju;Lee, You-Hwa;Cheong, Seon-Woo;Yoon, Chun-Sik
    • Journal of Environmental Science International
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    • v.19 no.8
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    • pp.1001-1012
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    • 2010
  • We investigated the toxic effects of tebuconazole on development in the African clawed frog, Xenopus laevis. To test the toxic effects, frog embryo teratogenesis assays using Xenopus were performed. Embryos were exposed to various concentrations of tebuconazole($0-100\;{\mu}M$). $LC_{100}$ for tebuconazole was $100\;{\mu}M$, and the $LC_{50}$ determined by probit analysis was $82.35\;{\mu}M$. The exposure to tebuconazole concentrations ${\geq}40\;{\mu}M$ resulted in 11 different types of severe external malformations including gut dysplasia. Histological examinations revealed various dysplasia in the eye, heart, liver, intestine, somatic muscle, and in the pronephric ducts. The tissue-specific toxic effects were investigated with an animal cap assay. Blood cells are generally induced at a high frequency by the combination of mSCF and activin A, however, the induction of blood cells was strongly inhibited by the addition of tebuconazole. Electron micrographs of tested embryos showed many of multivesicular bodies and dysplasia of photo-receptive cell, however, the somatic muscle degeneration was not severe. The gene expression of cultivated animal cap explants was investigated by reverse transcriptase-polymerase chain reaction and revealed that expression of the blood-specific marker, $\beta$ globin II and muscle-specific marker, muscle actin were more strongly inhibited than the neural-specific marker, XEn2.

Anti-fibrotic effects of L-2-oxothiazolidine-4-carboxylic acid via modulation of nuclear factor erythroid 2-related factor 2 in rats

  • Kim, In-Hee;Kim, Dae-Ghon;Hao, Peipei;Wang, Yunpeng;Kim, Seong-Hun;Kim, Sang-Wook;Lee, Seung-Ok;Lee, Soo-Teik
    • BMB Reports
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    • v.45 no.6
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    • pp.348-353
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    • 2012
  • L-2-Oxothiazolidine-4-carboxylic acid (OTC) is a cysteine prodrug that maintains glutathione in tissues. The present study was designed to investigate anti-fibrotic and anti-oxidative effects of OTC via modulation of nuclear factor erythroid 2-related factor 2 (Nrf2) in an in vivo thioacetamide (TAA)-induced hepatic fibrosis model. Treatment with OTC (80 or 160 mg/kg) improved serum liver function parameters and significantly ameliorated liver fibrosis. The OTC treatment groups exhibited significantly lower expression of ${\alpha}$-smooth muscle actin, transforming growth factor-${\beta}1$, and collagen ${\alpha}1$ mRNA than that in the TAA model group. Furthermore, the OTC treatment groups showed a significant decrease in hepatic malondialdehyde level compared to that in the TAA model group. Nrf2 and heme oxygenase-1 expression increased significantly in the OTC treatment groups compared with that in the TAA model group. Taken together, these results suggest that OTC restores the anti-oxidative system by upregulating Nrf2; thus, ameliorating liver injury and a fibrotic reaction.

Effect of Dietary Conjugated Linoleic Acid (CLA) on Abdominal Fat Deposition in Yellow-feather Broiler Chickens and Its Possible Mechanism

  • Zhou, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1760-1765
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    • 2008
  • A total of 60 one-day-old Yellow-feather broiler chickens were allotted into treatment and control groups. The treatment group was fed with the diet supplemented with 3% conjugated linoleic acid (CLA) for 48 d, while control group was fed with the diet supplemented with 3% rapeseed oil. Chickens were slaughtered in each group at the age of 49 d, and the blood and the abdominal adipose tissue were sampled. Serum cLeptin and serum cAdiponectin were measured by ELISA. The total RNA was extracted from adipose tissue to measure the abundance of the chicken growth hormone receptor (cGHR), insulin-like growth factor 1 (cIGF-1), insulin-like growth factor I receptor (cIGF-IR), peroxisome proliferator-activated receptor gamma ($cPPAR{\gamma}$), cAdiponectin and cAdipoIR mRNA by RT-PCR using ${\beta}$-actin as an internal standard. Results showed that the CLA decreased the abdominal fat index by 20.93% (p<0.05). The level of serum cLeptin but not serum cAdiponectin was significantly increased by CLA treatment (p<0.05). CLA down-regulated the relative abundance of cGH-R mRNA and $cPPAR{\gamma}$ mRNA in abdominal adipose tissue by 24.74% (p<0.05) and 66.52% (p<0.01) respectively. However, no differences were found between CLA treatment group and control group (p>0.05) in the relative abundance of cIGF-1, cIGF-IR, cAdiponectin, and cAdipoIR mRNA in abdominal adipose tissue. The data suggested that CLA inhibited abdominal fat deposition in broiler chicken may be determined by decreasing the GHR available for GH, and by inhibiting the differentiation of preadipocytes via down-regulation of $PPAR{\gamma}$, but independent of IGF and (or) GH-IGF pathway or adiponectin action.

Development of a New Duplex Real-Time Polymerase Chain Reaction Assay for Detection of Dicer in G. gallus

  • Ji, Xiaolin;Wang, Qi;Gao, Yulong;Wang, Yongqiang;Qin, Liting;Qi, Xiaole;Gao, Honglei;Wang, Xiaomei
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.630-636
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    • 2013
  • Recently, there has been a growing body of evidence showing that cellular microRNAs (miRNAs) are involved in virus-host interactions. Numerous studies have focused on analyses of the expression profiles of cellular miRNAs, but the expression patterns of Dicer, which is responsible for the generation of miRNAs, have only rarely been explored in Gallus gallus. We developed a duplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay for the relative quantification of the mRNAs of Dicer and ${\beta}$-actin in G. gallus. To apply this method, the expression of Dicer in avian cells after infection with avian leukosis virus subgroup J (ALV-J) was detected using our established duplex real-time RT-PCR. The duplex real-time RT-PCR assay is sufficiently sensitive, specific, accurate, reproducible, and cost-effective for the detection of Dicer in G. gallus. Furthermore, this study, for the first time, demonstrated that ALV-J can induce differential expression of Dicer mRNA in the ALV-J-infected cells.

STUDY ON THE EXPRESSION OF mRNA OF TUMOR NECROSIS FACTOR-α AND INTERLEUKIN-6 IN THE CELL LINES OF SQUAMOUS CELL CARCINOMA (구강 편평상피세포암종 세포주에서 Tumor Necrosis Factor-α와 Interleukin-6의 mRNA 발현에 관한 연구)

  • Ahn, Jin-Su;Kim, Kyung-Wook;Lee, Jae-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.6
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    • pp.535-542
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    • 2001
  • The purpose of this study was to examine the mRNA levels of TNF-${\alpha}$ and IL-6 in the cell lines of normal oral keratocyte and oral squamous cell carcinoma. Total RNA was extracted from these cell lines, observed under UV light, developed by radiographic films of PCR products via reverse transcriptase polymerase chain reaction(RT-PCR) amplication, and measured with densitometer. Each mRNA level of these cell lines divided by ${\beta}$-actin mRNA level was compared to that of normal control group. The results were as follows: 1. Higher mRNA expression of TNF-${\alpha}$ than IL-6 in the normal oral epithelial cell line. 2. In general, expression of mRNA of IL-6 appeared 3-4 times more in tumor cell lines than in control group. 3. mRNA expression of TNF-${\alpha}$ showed variable expression in tumor cell lines, unlike normal cell line. 4. There are no special connections between differentiation of oral cancer cell lines and mRNA expression of TNF-${\alpha}$ and IL-6. From the above results, expression of mRNA of IL-6 in the cell lines of squamous cell carcinoma used in this study has higher than the normal oral epithelial cell line, but there are no relationship between the differentiation of oral cancer cell lines and the expression of mRNA of TNF-${\alpha}$ and IL-6.

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Prognostic Relevance of Human Telomerase Reverse Transcriptase (hTERT) Expression in Patients with Gall Bladder Disease and Carcinoma

  • Deblakshmi, Raj Kumari;Deka, Manab;Saikia, Anjan Kumar;Sharma, Bir Kumar;Singh, Nidhi;Das, NN;Bose, Sujoy
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2923-2928
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    • 2015
  • Background: Gallbladder carcinoma (GBC) has been stated as an Indian disease, with the highest number of cases being reported from certain districts of northeast India, which has an ethnically distinct population. Unfortunately there are no scientific reports on the underlying molecular mechanisms associated with the pathogenesis of the disease from this region. Aim: The present study evaluated the role of differential expression of human telomerase reverse transcriptase (hTERT) in the development of gall bladder anomalies. Materials and Methods: Blood and tissue samples were collected from patients undergoing routine surgical resection for clinically proven cases of gallbladder disease {cholelithiasis (CL, n=50), cholecystitis (CS, n=40) and GBC (n=30) along with adjacent histopathologically proved non-neoplastic controls (n=15)} with informed consent. Whole blood was also collected from age and sex matched healthy controls (n=25) for comparative analysis. Differential hTERT mRNA expression was evaluated by semi-quantitative rt-PCR and real-time PCR based analysis using ${\beta}$-actin as an internal control. Evaluation of differential hTERT protein expression was studied by Western blot analysis and immunoflourescence. Statistical analysis for differential expression and co-relation was performed by SPSSv13.0 software. Results: Gallbladder anomalies were mostly prevalent in females. The hTERT mRNA and protein expression increased gradiently from normal