• Title/Summary/Keyword: G Proteins

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Optimization of PS-7 Production Process by Azotobacter indicus var. myxogenes L3 Using the Control of Carbon Source Composition (탄소원 조성 조절을 이용한 Azotobacter indicus var. myxogenes L3로부터 PS-7 생산 최적화)

  • Ra, Chae-Hun;Kim, Ki-Myong;Hoe, Pil-Woo;Lee, Sung-Jae;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.61-66
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    • 2008
  • The proteins in whey are separated and used as food additives. The remains (mainly lactose) are spray-dried to produce sweet whey powder, which is widely used as an additive for animal feed. Sweet whey powder is also used as a carbon source for the production of valuable products such as polysaccharides. Glucose, fructose, galactose, and sucrose as asupplemental carbon source were evaluated for the production of PS-7 from Azotobacter indicus var. myxogenes L3 grown on whey based MSM media. Productions of PS-7 with 2% (w/v) fructose and sucrose were 2.05 and 2.31g/L, respectively. The highest production of PS-7 was 2.82g/L when 2% (w/v) glucose was used as the carbon source. Galactose showed low production of PS-7 among the carbon sources tested. The effects of various carbon sources addition to whey based MSM medium showed that glucose could be the best candidate for the enhancement of PS-7 production using whey based MSM medium. To evaluate the effect of glucose addition to whey based media on PS-7 production, fermentations with whey and glucose mixture (whey 1, 2, 3%; whey 1% + glucose 1%, whey 1% + glucose 2% and glucose 2%, w/v) were carried out. Significant enhancement of PS-7 production with addition of 1% (w/v) and 2% (w/v) glucose in 1% (w/v) whey media was observed. The PS-7 concentration of 2% glucose added whey lactose based medium was higher than that of 1% glucose addition, however, the product yield $Y_{p/s}$ was higher in 1% glucose added whey lactose based MSM medium. Therefore, the optimal condition for the PS-7 production from the Azotobacter indicus var.myxogenes L3, was 1% glucose addition to 1% whey lactose MSM medium.

Protective effects of quality certified traditional Doenjang in Korea on TNF-α-induced vascular inflammation in human umbilical vein endothelial cells (혈관내피세포에서 TNF-α 자극에 의해 유도되는 혈관염증에 대한 전통식품 품질인증 된장의 효능 평가)

  • Kim, Eun-Ju;Jang, Yeon-Jeong;Kim, So-Young;Choi, Hye-Sun;Park, Shin-Young
    • Food Science and Preservation
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    • v.23 no.3
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    • pp.378-386
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    • 2016
  • Anti-atherogenic effects in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-stimulated human umbilical vein endothelial cells (HUVEC) are involved in the suppression of oxidative stress, cell adhesion molecules, and pro-inflammatory factors. This study investigated the vascular inflammation inhibitory activity of traditional Doenjang plays a key role in the pathogenesis and progression of atherosclerosis. The protective effects of Korean Deonjang was investigated on the expression of cell adhesion molecules (CAMs) in tumor necrosis factor (TNF)-${\alpha}$-induced human umbilical vascular endothelial cells (HUVECs). Deonjang extracts (20, 50, $100{\mu}g/mL$) decreased the expression of 20 ng/mL TNF-${\alpha}$-induced vascular cell adhesion molecule (VCAM)-1 intracellular adhesion molecule (ICAM)-1 proteins, and their corresponding mRNA levels. Nitric oxides (NO) produced by endothlial nitric oxides synthase (eNOS) dilated blood vessels, which had protective effects against platelet and leukocyte adhesion. While TNF-${\alpha}$-induced suppressed the production of nitric oxide in HUVECs, Doenjang restored NO production in HUVECs. In addition, Deonjang reduced the TNF-${\alpha}$-induced expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 mRNA levels. These results suggested that Doenjang can inhibited the production of cell adhesion molecules and inflammatory mediators, which could be a potential candidate for preventing atherosclerosis.

($P16^{ink4}$ Methylation in Squamous Cell Carcinoma of the Oral Cavity. (구강 편평세포암종에서 $P16^{ink4}$ 유전자의 Methylation에 대한 연구)

  • Kang, Gin-Won;Kim, Kyung-Wook;Lyu, Jin-Woo;Kim, Chang-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.2
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    • pp.164-173
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    • 2000
  • The p16 protein is a cyclin dependent kinase inhibitor that inhibits cell cycle progression from $G_1$ phase to S phase in cell cycle. Many p16 gene mutations have been noted in many cancer-cell lines and in some primary cancers, and alterations of p16 gene function by DNA methylation have been noticed in various kinds of cancer tissues and cell-lines. There have been a large body of literature has accumulated indicating that abnormal patterns of DNA methylation (both hypomethylation and hypermethylation) occur in a wide variety of human neoplasma and that these aberrations of DNA methylation may play an important epigenetic role in the development and progression of neoplasia. DNA methylation is a part of the inheritable epigenetic system that influences expression or silencing of genes necessary for normal differentiation and proliferation. Gene activity may be silenced by methylation of up steream regulatory regions. Reactivation is associated with demethylation. Although evidence or a high incidence of p16 alterations in a variety of cell lines and primary tumors has been reported, that has been contested by other investigators. The precise mechanisms by which abnormal methylation might contribute to carcinogenesis are still not fully elucidated, but conceivably could involve the modulation of oncogene and other important regulatory gene expression, in addition to creating areas of genetic instability, thus predisposing to mutational events causing neoplasia. There have been many variable results of studies of head and neck squamous cell carcinoma(HNSCC). This investigation was studied on 13 primary HNSCC for p16 gene status by protein expression in immunohistochemistry, and DNA genetic/epigenetic analyzed to determine the incidence, the mechanisms, and the potential biological significance of its Inactivation. As methylation detection method of p16 gene, the methylation specific PCR(MSP) is sensitive and specific for methylation of any block of CpG sites in a CpG islands using bisulfite-modified DNA. The genomic DNA is modified by treatment with sodium bisulfate, which converts all unmethylated cytosines to uracil(thymidine). The primers designed for MSP were chosen for regions containing frequent cytosines (to distinguish unmodified from modified DNA), and CpG pairs near the 5' end of the primers (to provide maximal discrimination in the PCR between methylated and unmethylated DNA). The two strands of DNA are no longer complementary after bisulfite treatment, primers can be designed for either modified strand. In this study, 13 paraffin embedded block tissues were used, so the fragment of DNA to be amplified was intentionally small, to allow the assessment of methylation pattern in a limited region and to facilitate the application of this technique to samlples. In this 13 primary HNSCC tissues, there was no methylation of p16 promoter gene (detected by MSP and automatic sequencing). The p16 protein-specific immunohistochemical staining was performed on 13 paraffin embedded primary HNSCC tissue samples. Twelve cases among the 13 showed altered expression of p16 proteins (negative expression). In this study, The author suggested that low expression of p16 protein may play an important role in human HNSCC, and this study suggested that many kinds of genetic mechanisms including DNA methylation may play the role in carcinogenesis.

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The Modulation of Radiosensitivity by Combined Treatment of Selective COX-2 Inhibitor, NS 398 and EGF Receptor Blocker AG 1478 in HeLa Cell Line (선택적 COX-2 억제제 NS 398과 EGF 수용체 차단제 AG 1478의 복합투여가 HeLa 세포주의 방사선 감수성에 미치는 영향)

  • Youn Seon Min;Oh Young Kee;Kim Joo Heon;Park Mi Ja;Seong In Ock;Kang Kimun;Chai Gyuyong
    • Radiation Oncology Journal
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    • v.23 no.1
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    • pp.51-60
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    • 2005
  • Purpose : Selective inhibition of multiple molecular targets may improve the antitumor activity of radiation. Two specific inhibitors of selective cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) were combined with radiation on the HeLa cell line. To investigate cooperative mechanism with selective COX-2 inhibitor and EGFR blocker, in vitro experiments were done. Materials and Methods : Antitumor effect was obtained by growth inhibition and apoptosis analysis by annexin V-Flous method. Radiation modulation effects were determined by the clonogenic cell survival assay. Surviving fractions at 2 Gy ($SF_2$) and dose enhancement ratio at a surviving fraction of 0.25 were evaluated. To investigate the mechanism of the modulation of radiosensitivity, the cell cycle analyses were done by flow cytometry. The bcl-2 and bax expressions were analyzed by western blot. Results : A cooperative effect were observed on the apoptosis of the HeLa ceil line when combination of the two drugs, AG 1478 and NS 398 with radiation at the lowest doses, apoptosis of $22.70\%$ compare with combination of the one drug with radiation, apoptosis of $8.49\%$. In cell cycle analysis, accumulation of cell on $G_0/G_l$ phase and decrement of S phase fraction was observed from 24 hours to 72 hours after treatment with radiation, AG 1478 and NS 398. The combination of NS 398 and AG 1478 enhanced radiosensitivity on a concentration-dependent manner in HeLa cells with dose enhancement ratios of 3.00 and $SF_2$ of 0.12 but the combination of one drug with radiation was not enhanced radlosensitivity with dose enhancement ratios of 1.12 and SF2 of 0.68 (p=0.005). The expression levels of bcl-2 and bax were reduced when combined with AG 1478 and NS 398. Conclusion : Our results indicate that the selective COX-2 inhibitor and EGFR blocker combined with radiation have potential additive or cooperative effects on radiation treatment and may act through various mechanisms including direct inhibition of tumor cell proliferation, suppression of tumor cell cycle progression and inhibition of anti-apoptotic proteins.

Anti-inflammatory Effect of Ethanol Extract from Sargassum fulvellum on Lipopolysaccharide Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀조직에 대한 참모자반 (Sargassum fulvellum) 에탄올 추출물의 항염증 효과)

  • Jeong, Da-Hyun;Kim, Koth-Bong-Woo-Ri;Kim, Min-Ji;Kang, Bo-Kyeong;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Ahn, Na-Kyung;Choi, Yeon-Uk;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1158-1165
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    • 2014
  • This study investigated the anti-inflammatory effects of Sargassum fulvellum ethanol extract (SFEE) on the lipopolysaccharide (LPS)-induced inflammatory response. SFEE remarkably suppressed production of NO and pro-inflammatory cytokines (IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ at 50 and $100{\mu}g/mL$. There were no cytotoxic effects on proliferation of macrophages treated with SFEE compared to the control. SFEE reduced expression of iNOS and COX-2 proteins in a dose-dependent manner. The formation of edema in mouse ears was reduced at the highest dose tested compared to the control. Moreover, in the acute toxicity test, no mortality occurred in mice administered 5,000 mg/kg body weight of SFEE over the 2-week observation period. These results suggest that SFEE may have significant effects on inflammatory factors and be a potential anti-inflammatory therapeutic material.

Bioactive Characterization of Bacillus thuriniensis subsp. kurstaki CAB133 Isolated from Domestic Soil (국내 토양으로 분리된 Bacillus thuriniensis subsp. kurstaki CAB133균주의 생물학적 특성)

  • Choi, Su-Yeon;Cho, Min-Su;Kim, Tae-Hwan;Kim, Jin-Su;Pack, Seung-Kyung;Youn, Young-Nam;Hong, Soon-Sung;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.47 no.2
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    • pp.175-184
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    • 2008
  • To screen highly active Bacillus thuringiensis isolates against Spodoptera litura (Lepidoptera, Noctuidae), 46 B. thuringiensis was isolated from 115 samples obtained from several crop soils. Especially, B. thuringiensis subsp. kurstaki CAB133 and CAB162 isolates showed 100% mortality against S. litura. $LD_{50}$ values of CAB 133, CAB162 and HD-1 strains of B. thuringiensis subsp. kurstaki were 0.089, 3.144 and $0.513{\mu}g/ml$ against 2nd larva of S. litura, respectively. The weight of 3rd larva of S. litura which were fed crystal inclusion protein $(1.267{\mu}g/ml)$ with B. thuringiensis subsp. kurstaki CAB133 was about 30 times lass than control group. CAB133 and CAB 162 strains of B. thuringiensis subsp. kurstaki which were taken a highly toxity against S. litura were analyzed by SDS-PAGE, and estimated the molecular weight of the Cry proteins. Their serological identification by H serotypes were showed B. thuringiensis subsp. kurstaki (3abc) type.

Effects of Autoclave and Microwave Treatments on the Antigenicity of Gliadin in Medium Wheat Dough (가압가열 및 microwave에 의한 중력분 반죽 gliadin의 항원성 변화)

  • Kwak, Ji-Hee;Kim, Koth-Bong-Woo-Ri;Lee, Chung-Jo;Kim, Min-Ji;Kim, Dong-Hyun;SunWoo, Chan;Jung, Seul-A;Kim, Hyun-Jee;Choi, Jung-Su;Kim, Seong-Won;Ahn, Dong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.44 no.1
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    • pp.55-60
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    • 2012
  • The aim of this study was to determine the optimal physical treatment to reduce the antigenicity of gliadin in wheat dough. Medium wheat dough was treated with an autoclave (5, 10, 30, and 50 min at $121^{\circ}C$, 1 atm), a microwave (1, 5, and 10 min) or both (10, 30, and 50 min/5, 10 min). The proteins in the dough extracts were analyzed by SDSPAGE and the binding ability of anti-gliadin IgG to gliadin was examined by ci-ELISA and immunoblotting. Results showed that the ability of anti-gliadin IgG to bind to gliadin in wheat dough treated with an autoclave alone or in combination with a microwave was decreased. Especially, it declined to ~77% after autoclaving for 30 min and 35% after both autoclaving for 50 min and microwaving for 5 min. In addition, the intensity of gliadin bands in SDS-PAGE were weakened and anti-gliadin IgG did not recognize gliadin in immunoblotting. However, microwaving alone did not affect the antigenicity of gliadin in wheat dough. These results indicate that autoclaving may affect the reduction of the antigenicity of gliadin in medium wheat dough. Moreover, autoclaving in combination with microwaving is more effective for reducing the antigenicity of wheat dough.

Inhibition of Pancreatic Lipase Activity and Adipocyte Differentiation in 3T3-L1 Cells Treated with Purple Corn Husk and Cob Extracts (자색옥수수 포엽과 속대 추출물의 리파아제 저해활성 및 3T3-L1 지방전구세포에서의 지방분화 억제효과)

  • Lee, Ki Yeon;Hong, Soo Young;Kim, Tae Hee;Kim, Jai Eun;Park, A-Reum;Noh, Hee Sun;Kim, Si Chang;Park, Jong Yeol;Ahn, Mun Seob;Jeong, Won Jin;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.131-139
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    • 2018
  • Our review begins with the maize hybrid for grain, called 'Seakso 1,' which was developed in 2008 by the Gangwon Agricultural Research and Extension Services in Korea, and subsequently registered in 2011. In this study, we aimed to investigate the lipid metabolic enzyme activity and inhibitory effect on the adipocyte differentiation, in 3T3-L1 cells of the identified Seakso 1 corn husk and cob extracts (EHCS). We investigated the pancreatic lipase inhibitory effect and anti-adipogenic effect of EHCS.The lipid accumulation and adipocyte differentiation were measured by the procedure of Oil Red O staining, Real-time PCR and the Western blot analysis. The pancreatic lipase inhibitory activity of EHCS was measured at higher levels than those of the positive control (orlistat) at 100, 500, and $1,000{\mu}g/mL$. In particular, EHCS was noted as being significantly inhibited and including a measured adipocyte differentiation and lipid accumulation, when treated during the adipocyte differentiation process in 3T3-L1 cells. Based on the Oil Red O staining, EHCS inhibited lipid accumulation at 19.19%, 33.30% at $1000{\mu}g/mL$, $2000{\mu}g/mL$, respectively. The real-time PCR and Western blot analysis showed that EHCS significantly decreased in the mRNA expression and protein level of obesity-related factors, such as peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). This study potentially suggests that the Saekso 1 corn husk and cob extracts may improve lipid metabolism and reduce lipid accumulation.

EFFECT OF ENAMEL MATRIX DERIVATIVE (EMD, $EMDOGAIN^{(R)}$) ON THE DIFFERENTIATION OF CULTURED HUMAN PERIODONTAL LIGAMENT CELLS AND MESENCHYMAL STEM CELLS (배양된 사람 치주인대세포와 골수유래간엽줄기세포의 분화에 미치는 법랑기질유도체 (Enamel Matrix Derivative, EMD)의 영향)

  • Park, Sang-Gyu;Jue, Seong-Suk;Kwon, Yong-Dae;Choi, Byung-Joon;Kim, Young-Ran;Lee, Baek-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.4
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    • pp.281-286
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    • 2009
  • Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

Expression of CsRCI2s by NaCl stress reduces water and sodium ion permeation through CsPIP2;1 in Camelina sativa L.

  • Kim, Hyun-Sung;Lim, Hyun-Gyu;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.194-194
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    • 2017
  • Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

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