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Sample records for macc1 specific shrna

  1. 下调肝星状细胞 MACC1对胃癌细胞迁移侵袭能力的影响%Effect of down -regulating MACC1 by shRNA on invasion and migration of gastric carci-noma cells

    Institute of Scientific and Technical Information of China (English)

    袁园; 朱正秋

    2017-01-01

    Objective:By in vitro experiment to explore the regulatory effect of MACC1 gene silencing in HSC expressα-SMA,MMP -2,MMP -9 and HGF,and its effects on ability of migration and invasion of co -cultured gastric carcinoma cells.Methods:To construct targeting MACC1 gene eukaryotic expression interference plasmid (MACC1 -shRNA)and non -specific irrelevant interference plasmid (shNC),transient transfect them into HSC with lipofectamine -2000,named low -expression group(shRNA)and negative control group (NC)accordingly,non-transfected HSC treated as a blank control group(Blank -Ctrl).The above three groups of cells were cultured in accordance with routinel,the expression levels of MACC1,α-SMA,HGF,MMP -2 and MMP -9 in human hepatic stellate cells were detected by Western blot and RT -PCR.Three groups of cells were co -cultured with MGC803 gastric carcinoma cells in vitro,the capabilities of migration and invasion were measured by transwell assay.Results:Western -Blot results showed that the expression of MACC1,α-SMA,HGF,MMP -2 and MMP -9 protein in HSC was significantly lower than that in the negative group and blank control group after transfection of MACC1 -shRNA (P 0.05).Conclusion:Silencing MACC1 in hepatic stellate cells, can inhibit the expression of MMP -2,MMP -9 and HGF,and decrease the migration and invasion of gastric cancer cells in co -culture.%目的:通过体外实验探讨肝星状细胞(hepatic stellate cell,HSC)MACC1基因沉默后调节肝星状细胞激活标志物α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、肝细胞生长因子(hepatocyte growth fac-tor,HGF)、基质金属蛋白酶2(matrix metalloproteinase -2,MMP -2)、基质金属蛋白酶9(matrix metalloprotein-ase -9,MMP -9)的表达,以及对共培养胃癌细胞迁移侵袭能力的影响。方法:利用 RNA 干扰技术,构建针对MACC1基因的真核表达干涉质粒(MACC1-shRNA)、非特异性无关

  2. MACC1基因siRNA对肺癌SBC-5细胞增殖和迁移的影响%Influence of MACC1 Gene expression on the proliferation and migration of SBC-5 lung cancer cells

    Institute of Scientific and Technical Information of China (English)

    杨淑慧; 龙敏; 王希; 林芳; 张惠中

    2011-01-01

    目的:探讨MACC1(metastasis-associated in colon cancer l)基因特异性siRNA(small interfering RNA)对肺癌细胞系SBC-5体外增殖和迁移的影响.方法:化学合成MACC1基因特异性siRNA,阳离子脂质体LipofectamineTM 2000转染肺癌细胞系SBC-5细胞,RT-PCR和Western blot方法检测转染后SBC-5细胞中MACC1基因和蛋白的表达情况;MTT法检测细胞的生长增殖状况;划痕试验观察细胞迁移能力的改变.结果:MACC1基因特异性siRNA转染细胞后,SBC-5细胞MACC1的mRNA和蛋白表达水平明显降低,MTT试验和划痕试验观察到转染MACC1-siRNA的SBC-5细胞增殖、迁移能力明显减弱.结论:MACC1基因特异性siRNA可以明显抑制肺癌SBC-5细胞增殖和迁移,MACC1可能成为肺癌治疗的新靶点.%Objective: To investigate the influence of siRNA targeting metastasis - associated in colon cancer Ⅰ ( MACC1 ) gene on proliferation and migration of human lung cancer cell line SBC - 5 in vitro. Methods : The siRNA targeting MACC1 gene and negative control siRNA were chemically synthesized, and were then transiently transfected into human lung cancer cell line SBC - 5 hy LipofectamineTM 2000. Expression of MACC1 mRNA and protein in siRNA transfected cells were examined by RT - PCR and Western blotting analysis. Proliferation and migration of siRNA transfected SBC - 5 cells were examined by MTT and Wound healing assay. Results : Transfection of specific siRNA targeting MACC1 into SBC - 5 cells inhihited MACC1 gene expression and both mRNA and protein expression levels declined markedly. siRNA transfected SBC - 5 cells showed decreased ability of proliferation and migration. Conclusion : The specific siRNA targeting MACC1 gene can significantly inhibit proliferation and migration of SBC - 5 cells, MACC1 may serve as a potential target for gene therapy of lung cancer.

  3. The Generation of Eukaryotic Expression Vectors of shRNA Specific for Stat6

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    1 Introduction Since RNA interference (RNAi) was first discovered in plant and the nematode Caenorhabditis elegans in 1998~([1]), it quickly became a powerful and indispensable tool in the molecular toolkit. Small interfering RNA (siRNA) that binds to specific endogenous mRNA and induce their degradation is now widely employed to inhibit protein expression at post-transcriptional level. Recently, RNAi techniques have employed short hairpin RNA (shRNA) instead of siRNA because shRNA can generate siRNA in cel...

  4. Identification of MACC1 as a novel prognostic marker in hepatocellular carcinoma

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    Lu Canliang

    2011-09-01

    Full Text Available Abstract Background Metastasis-associated in colon cancer-1 (MACC1 is a newly identified gene that plays a role in colon cancer metastasis through upregulation of c-MET proto-oncogene (c-MET. However, the value of MACC1 as a potential biomarker for hepatocellular carcinoma (HCC remains unknown. Methods MACC1 mRNA expression in 128 HCC tissues was examined by quantitative polymerase chain reaction. To show the potential correlation of MACC1 and c-MET, c-MET was also analysed. Results MACC1 was more highly expressed in HCC than in non-HCC tissues (P = 0.009. High MACC1 expression was significantly increased in cases with high alpha fetoprotein (AFP (P = 0.025. A positive correlation was found between MACC1 and c-MET mRNAs (r = 0.235, P = 0.009. Both univariate and multivariate analyses revealed that MACC1 expression was associated with overall survival (OS and disease-free survival (DFS. Moreover, stratified analysis showed that tumour-node-metastasis (TNM stage I patients with high MACC1 levels had shorter OS and DFS than those with low MACC1. Conclusions MACC1 may identify low- and high-risk individuals with HCC and be a valuable indicator for stratifying the prognosis of TNM stage I patients. MACC1 may serve as a novel biomarker for HCC.

  5. MACC1 as a prognostic biomarker for early-stage and AFP-normal hepatocellular carcinoma.

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    Chan Xie

    Full Text Available BACKGROUND: The metastasis-associated in colon cancer 1 gene (MACC1 has been found to be associated with cancer development and progression. The aim of this study was to investigate the prognostic value of MACC1 in early-stage and AFP-normal hepatocellular carcinoma (HCC. METHODS: mRNA and protein levels of MACC1 expression in one normal liver epithelial cells THLE3 and 15 HCC cell lines were examined using reverse transcription-PCR and Western blot. MACC1 expression was also comparatively studied in 6 paired HCC lesions and the adjacent non-cancerous tissue samples. Immunohistochemistry was employed to analyze MACC1 expression in 308 clinicopathologically characterized HCC cases. Statistical analyses were applied to derive association between MACC1 expression scores and clinical staging as well as patient survival. RESULTS: Levels of MACC1 mRNA and protein were higher in HCC cell lines and HCC lesions than in normal liver epithelial cells and the paired adjacent noncancerous tissues. Significant difference in MACC1 expression was found in patients of different TNM stages (P<0.001. Overall survival analysis showed that high MACC1 expression level correlated with lower survival rate (P = 0.001. Importantly, an inverse correlation between MACC1 level and patient survival remained significant in subjects with early-stage HCC or with normal serum AFP level. CONCLUSIONS: MACC1 protein may represent a promising biomarker for predicting the prognosis of HCC, including in early-stage and AFP-normal patients.

  6. 肿瘤转移的一个靶向治疗位点MACC1%A target site for the treatment of tumor meatastasis: MACC1

    Institute of Scientific and Technical Information of China (English)

    贺志云; 白志刚; 张忠涛

    2014-01-01

    Metastasis-associated in colon cancer-1 (MACC1) is a recently discovered gene associated with colon cancer metastasis,there is significant relationship indicated from some studies between MACC1 and different malignant tumors.It may play an important role in the regulation of tumors metastasis.This article reviewed the expression and regulating function of MACC1 in different cancers including colorectal cancer,hepatic cancer,gastric cancer,lung cancer,ovarian cancer,breast cancer,and so on.It may offer clues to find a new target for target treatment of cancer metastasis.%结肠癌转移相关基因-1(MACC1)是新近发现的一个与结肠癌密切相关的基因,研究表明,MACC1与多种恶性肿瘤的转移和复发有密切的相关性,可能在肿瘤转移的共同通路中起着至关重要的调控作用.本研究就MACC1在结肠癌、肝癌、胃癌、肺癌、卵巢癌和乳腺癌等多种恶性肿瘤中的表达与功能调控等方面进行综述,以期为肿瘤转移调控的靶向治疗提供一个新的方向.

  7. Prognostic Value of MACC1 in Digestive System Neoplasms: A Systematic Review and Meta-Analysis.

    Science.gov (United States)

    Wu, Zhenzhen; Zhou, Rui; Su, Yuqi; Sun, Li; Liao, Yulin; Liao, Wangjun

    2015-01-01

    Metastasis associated in colon cancer 1 (MACC1), a newly identified oncogene, has been associated with poor survival of cancer patients by multiple studies. However, the prognostic value of MACC1 in digestive system neoplasms needs systematic evidence to verify. Therefore, we aimed to provide further evidence on this topic by systematic review and meta-analysis. Literature search was conducted in multiple databases and eligible studies analyzing survival data and MACC1 expression were included for meta-analysis. Hazard ratio (HR) for clinical outcome was chosen as an effect measure of interest. According to our inclusion criteria, 18 studies with a total of 2,948 patients were identified. Pooled HRs indicated that high MACC1 expression significantly correlates with poorer OS in patients with digestive system neoplasms (HR = 1.94; 95% CI: 1.49-2.53) as well as poorer relapse-free survival (HR = 1.94, 95% CI: 1.33-2.82). The results of subgroup studies categorized by methodology, anatomic structure, and cancer subtype for pooled OS were all consistent with the overall pooled HR for OS as well. No publication bias was detected according to test of funnel plot asymmetry and Egger's test. In conclusion, high MACC1 expression may serve as a prognostic biomarker to guide individualized management in clinical practice for digestive system neoplasms.

  8. Prognostic Value of MACC1 and c-met Expressions in Non-small Cell Lung Cancer

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    Xingsheng HU

    2012-07-01

    Full Text Available Background and objective It has been proven that metastasis-associated in colon cancer 1 (MACC1 is a new gene that is related to the invasion and metastasis of tumors. MACC1 also regulates c-met expression. The aim of this study is to explore the expressions of MACC1 and hepatocyte growth factor receptor (c-met, and its relationship with invasion, metastasis, and prognosis of non-small cell lung cancer (NSCLC. Methods MACC1 and c-met expressions were detected in 103 cases of NSCLC and 40 cases of neighboring normal lung cancer tissue using immunohistochemistry. Results MACC1 and c-met expressions were significantly higher in lung cancer tissues than that in neighboring normal tissue (P<0.001. MACC1 and c-met expressions were associated with poor differentiation, advanced T stages, lymph node metastasis, and advanced TNM stages (P<0.05 of NSCLC, but not with sex, age, smoking, and histological classification (P>0.05. In addition, a positive correlation between MACC1 and c-met expressions was observed (r=0.403, P<0.001. The result from the Kaplan-Meier survival analysis showed that the five-year survival rate in patients with positive MACC1 and c-met expressions was remarkanly lower than that in patients with negative expressions (P<0.05. The result from the Cox regression analysis showed that MACC1 expression was an independent prognostic factor for NSCLC (P=0.026. Conclusion MACC1 and c-met have an important function in the differentiation, invasion, and metastasis of NSCLC. MACC1 and c-met have poor prognosis in patients with NSCLC. Moreover, MACC1 expression is an independent prognostic factor for NSCLC.

  9. Targeting MACC1 by RNA interference inhibits proliferation and invasion of bladder urothelial carcinoma in T24 cells.

    Science.gov (United States)

    Xu, Song-Tao; Ding, Xiang; Ni, Qing-Feng; Jin, Shao-Ju

    2015-01-01

    The purpose of this article is to research on whether MACC1 can serve as a potential target for gene therapy of human bladder urothelial carcinoma (BUC). In this study, the expression of MACC1 gene was knocked down by RNA interference (RNAi) in the T24 cell (human BUC cell). The transcription level of MACC1 was detected by RT-PCR. Activities of MACC1, caspase-3, caspase-8, Bax and Met (mesenchymal-epithelial transition factor) protein were measured by Western blot. The cell proliferation and apoptosis were detected by MTT and flow cytometry. The cell's invasion ability was performed on Matrigel transwell assay. We also detect MMP2 (metalloproteinase-2) proteins by ELISA. The results showed that the level of MACC1 mRNA and protein was significantly reduced after RNAi. MTT assay showed that the proliferation of T24 cell was decreased due to RNA interference. Apoptosis studies also showed that MACC1 gene interference in T24 loses its anti-apoptotic effects. The expression of apoptosis proteins (Caspase-3, Caspase-8 and Bax) increased significantly due to the MACC1 RNAi. The level of Met protein was down-regulated obviously due to RNAi. Transwell assay showed that invasion abilities of T24 cells were reduced obviously due to MACC1 RNAi. Further studies showed that the secretion of MMP-2 was reduced by RNAi. It can conclude that the ability of proliferation and invasion in T24 cells can be inhibited by RNAi-targeting MACC1. As a result, MACC1 can serve as a potential target for gene therapy of human bladder urothelial carcinoma.

  10. Prognostic significance of metastasis associated in colon cancer 1 (MACC1 expression in patients with gallbladder cancer

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    Lijian Chen

    2014-01-01

    Full Text Available Background: The clinical significance of metastasis associated in colon cancer 1 (MACC1, in human gallbladder cancer, is not yet established. This study was performed to assess the expression of MACC1 in benign and malignant gallbladder lesions, and to assess its clinicopathological significance. Materials and Methods: Tissue samples from resected gallbladder cancer (n = 70 and cholelithiasis (n = 70 were evaluated for MACC1 expression by immunohistochemical staining. Their expression was correlated with different clinicopathological parameters. Results: Cytoplasmic MACC1 expression was significantly higher (58.6% in gallbladder cancer than in chronic cholecystitis (27.1%, P 0.05. The univariate Kaplan-Meier analysis showed that a positive MACC1 expression was associated with decreased overall survival (P < 0.001. The multivariate Cox regression analysis showed that MACC1 expression and the histopathological subtypes were independent risk factors for disease-free survival. Conclusion: The expression of MACC1 might be closely related to carcinogenesis, clinical biological behaviors, and prognosis of gallbladder adenocarcinoma.

  11. SNPs in the coding region of the metastasis-inducing gene MACC1 and clinical outcome in colorectal cancer

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    Schmid Felicitas

    2012-07-01

    Full Text Available Abstract Background Colorectal cancer is one of the main cancers in the Western world. About 90% of the deaths arise from formation of distant metastasis. The expression of the newly identified gene metastasis associated in colon cancer 1 (MACC1 is a prognostic indicator for colon cancer metastasis. Here, we analyzed for the first time the impact of single nucleotide polymorphisms (SNPs in the coding region of MACC1 for clinical outcome of colorectal cancer patients. Additionally, we screened met proto-oncogene (Met, the transcriptional target gene of MACC1, for mutations. Methods We sequenced the coding exons of MACC1 in 154 colorectal tumors (stages I, II and III and the crucial exons of Met in 60 colorectal tumors (stages I, II and III. We analyzed the association of MACC1 polymorphisms with clinical data, including metachronous metastasis, UICC stages, tumor invasion, lymph node metastasis and patients’ survival (n = 154, stages I, II and III. Furthermore, we performed biological assays in order to evaluate the functional impact of MACC1 SNPs on the motility of colorectal cancer cells. Results We genotyped three MACC1 SNPs in the coding region. Thirteen % of the tumors had the genotype cg (rs4721888, L31V, 48% a ct genotype (rs975263, S515L and 84% a gc or cc genotype (rs3735615, R804T. We found no association of these SNPs with clinicopathological parameters or with patients’ survival, when analyzing the entire patients’ cohort. An increased risk for a shorter metastasis-free survival of patients with a ct genotype (rs975263 was observed in younger colon cancer patients with stage I or II (P = 0.041, n = 18. In cell culture, MACC1 SNPs did not affect MACC1-induced cell motility and proliferation. Conclusion In summary, the identification of coding MACC1 SNPs in primary colorectal tumors does not improve the prediction for metastasis formation or for patients’ survival compared to MACC1 expression analysis alone. The ct genotype (rs

  12. Over-expression of Metastasis-associated in Colon Cancer-1 (MACC1)Associates with Better Prognosis of Gastric Cancer Patients

    Institute of Scientific and Technical Information of China (English)

    Shao-hua Ge; Jia-fu Ji; Xiao-jiang Wu; Xiao-hong Wang; Xiao-fang Xing; Lian-hai Zhang; Yu-bing Zhu; Hong Du; Bin Dong; Ying Hu

    2011-01-01

    Objective: The aim of this study was to detect metastasis-associated in colon cancer-1 (MACC1) expression in Chinese gastric cancer and analyze the relationship between MACC1 expression and postoperative survival. Methods: The expression of MACC1 and c-MET protein in a sample of 128 gastric cancer tissues was detected by immunohistochemistry. A retrospective cohort study on the prognosis was carried out and data were collected from medical records. Results: The positive rate of MACC1 protein expression in gastric cancer was 47.66%, higher than that in adjacent noncancerous mucosa (P<0.001). MACC1 protein expression was not related to the clinicopathological variables involved. Kaplan-Meier analysis revealed that the survival of MACC1 positive group tended to be better than that of MACC1 negative group, particularly in patients with stage Ⅲ carcinoma (P=0.032). Cox regression analysis revealed that MACC1 protein over-expression in gastric cancer tended to be a protective factor with hazard ratio of 0.621 (P=0.057). Immunohistochemical analysis showed that the positive rate of c-MET protein expression was much higher in cases with positive MACC1 expression in gastric cancer (P=0.002), but P53 expression was not associated with MACC1 expression. Conclusion: MACC1 over-expression implies better survival and may be an independent prognostic factor for gastric cancer in Chinese patients.

  13. A Small Indel Mutant Mouse Model of Epidermolytic Palmoplantar Keratoderma and Its Application to Mutant-specific shRNA Therapy.

    Science.gov (United States)

    Lyu, Ya-Su; Shi, Pei-Liang; Chen, Xiao-Ling; Tang, Yue-Xiao; Wang, Yan-Fang; Liu, Rong-Rong; Luan, Xiao-Rui; Fang, Yu; Mei, Ru-Huan; Du, Zhen-Fang; Ke, Hai-Ping; Matro, Erik; Li, Ling-En; Lin, Zhao-Yu; Zhao, Jing; Gao, Xiang; Zhang, Xian-Ning

    2016-03-22

    Epidermolytic palmoplantar keratoderma (EPPK) is a relatively common autosomal-dominant skin disorder caused by mutations in the keratin 9 gene (KRT9), with few therapeutic options for the affected so far. Here, we report a knock-in transgenic mouse model that carried a small insertion-deletion (indel) mutant of Krt9, c.434delAinsGGCT (p.Tyr144delinsTrpLeu), corresponding to the human mutation KRT9/c.500delAinsGGCT (p.Tyr167delinsTrpLeu), which resulted in a human EPPK-like phenotype in the weight-stress areas of the fore- and hind-paws of both Krt9(+/mut) and Krt9(mut/mut) mice. The phenotype confirmed that EPPK is a dominant-negative condition, such that mice heterozygotic for the K9-mutant allele (Krt9(+/mut)) showed a clear EPPK-like phenotype. Then, we developed a mutant-specific short hairpin RNA (shRNA) therapy for EPPK mice. Mutant-specific shRNAs were systematically identified in vitro using a luciferase reporter gene assay and delivered into Krt9(+/mut) mice. shRNA-mediated knockdown of mutant protein resulted in almost normal morphology and functions of the skin, whereas the same shRNA had a negligible effect in wild-type K9 mice. Our results suggest that EPPK can be treated by gene therapy, and this has significant implications for future clinical application.

  14. SPON2, a newly identified target gene of MACC1, drives colorectal cancer metastasis in mice and is prognostic for colorectal cancer patient survival.

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    Schmid, F; Wang, Q; Huska, M R; Andrade-Navarro, M A; Lemm, M; Fichtner, I; Dahlmann, M; Kobelt, D; Walther, W; Smith, J; Schlag, P M; Stein, U

    2016-11-17

    MACC1 (metastasis associated in colon cancer 1) is a prognostic biomarker for tumor progression, metastasis and survival of a variety of solid cancers including colorectal cancer (CRC). Here we aimed to identify the MACC1-induced transcriptome and key players mediating the MACC1-induced effects in CRC. We performed microarray analyses using CRC cells ectopically overexpressing MACC1. We identified more than 1300 genes at least twofold differentially expressed, including the gene SPON2 (Spondin 2) as 90-fold upregulated transcriptional target of MACC1. MACC1-dependent SPON2 expression regulation was validated on mRNA and protein levels in MACC1 high (endogenously or ectopically) and low (endogenously or by knockdown) expressing cells. Chromatin immunoprecipitation analysis demonstrated the binding of MACC1 to the gene promoter of SPON2. In cell culture, ectopic SPON2 overexpression induced cell viability, migration, invasion and colony formation in endogenously MACC1 and SPON2 low expressing cells, whereas SPON2 knockdown reduced proliferative, migratory and invasive abilities in CRC cells with high endogenous MACC1 and SPON2 expression. In intrasplenically transplanted NOD/SCID mice, metastasis induction was analyzed with control or SPON2-overexpressing CRC cells. Tumors with SPON2 overexpression induced liver metastasis (vs control animals without any metastases, P=0.0036). In CRC patients, SPON2 expression was determined in primary tumors (stages I-III), and survival time was analyzed by Kaplan-Meier method. CRC patients with high SPON2 expressing primary tumors demonstrated 8 months shorter metastasis-free survival (MFS) compared with patients with low SPON2 levels (P=0.053). Combining high levels of SPON2 and MACC1 improved the identification of high-risk patients with a 20-month shorter MFS vs patients with low biomarker expression. In summary, SPON2 is a transcriptional target of the metastasis gene MACC1. SPON2 induces cell motility in vitro and CRC

  15. Targeting cleavage and polyadenylation specific factor 1 via shRNA inhibits cell proliferation in human ovarian cancer

    Indian Academy of Sciences (India)

    BEIGUANG ZHANG; YING LIU; DONGHUI LIU; LEI YANG

    2017-09-01

    Cleavage and polyadenylation specificity factor 1 (CPSF1), a member of CPSF complex, has been reported to play a keyrole in pre-mRNA 30-end formation, but its possible role in ovarian cancer remains unclear. In the present study, we foundthe mRNA level of CPSF1 was overexpressed in ovarian cancer tissues using Oncomine Cancer Microarray database. Thenthe loss-of-function assays, including CCK-8, colony formation and flow cytometry assays, were performed to determinethe effects of CPSF1 on cell viability, proliferation, cell cycle and apoptosis of human ovarian cancer cell lines (SKOV-3and OVCAR-3). The results indicated that depletion of CPSF1 suppressed cell viability, impaired colony formation ability,induced cell cycle arrest at G0/G1 phase and promoted cell apoptosis in ovarian cancer cells. Furthermore, knockdown ofCPSF1 upregulated the expression of cleaved caspase-3 and PARP and downregulated CDK4/cyclin D1 expression. Thesedata suggested that CPSF1 could promote ovarian cancer cell growth and proliferation in vitro and its depletion might serveas a potential therapeutic target for human ovarian cancer.

  16. Expression of P2X7R in breast cancer tissue and the induction of apoptosis by the gene-specific shRNA in MCF-7 cells

    OpenAIRE

    Tan, Chao; Han, Li; Zou, Lili; Luo, Chunhua; Liu, Aihua; SHENG, XIEJING; XI, DEE

    2015-01-01

    The aim of the present study was to investigate the effects of P2X7R short hairpin (sh)RNA on the proliferation and apoptosis of MCF-7 cells, and to detect the expression of P2X7R in breast cancer and MCF-7 cells. In order to detect the expression of P2X7R in normal breast and breast cancer tissues, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blot analysis and immunohistochemistry were performed. A P2X7-targeted shRNA sequence and a scrambled sequence were ...

  17. Relationship between pathological characteristics of prostate cancer and MACC1, c-Met, Apaf-1 as well as Caspase-9 expression in tumor tissue

    Institute of Scientific and Technical Information of China (English)

    Hang-Yu Ai; Xue-De Qiu

    2016-01-01

    Objective:To study the MACC1, c-Met, Apaf-1 and Caspase-9 expression in prostate cancer tissue and their relationship with the pathological characteristics of tumor.Methods:Prostate cancer and benign prostatic hyperplasia patients who received surgical treatment in our hospital from May 2015 to March 2016 were selected as the research subjects, prostate cancer tissue and benign prostatic hyperplasia tissue were collected during surgery to determine MACC1, c-Met, Apaf-1 and Caspase-9 expression, and serum specimens were collected to determine miR-let7i, -32, -128, -196a and -218 expression levels.Results: mRNA content of MACC1 and c-Met in prostate cancer tissue were significantly higher than those in benign prostatic hyperplasia tissue while mRNA content of Apaf-1 and Caspase-9 were significantly lower than those in benign prostatic hyperplasia tissue, and the higher the Gleason grading and the higher the Whitmore-Prout staging, the higher the mRNA content of MACC1 and c-Met in prostate cancer tissue and the lower the mRNA content of Apaf-1 and Caspase-9; serum miR-32, miR-128 and miR-196a expression levels in prostate cancer patients were significantly higher than those in patients with benign prostatic hyperplasia and negatively correlated with the mRNA content of Apaf-1 and Caspase-9, and the expression levels of miR-let7i and miR-218 were significantly lower than those in patients with benign prostatic hyperplasia and negatively correlated with MACC1 and c-Met.Conclusion: High MACC1 and c-Met expression and low Caspase-9 and Apaf-1 expression are related to the occurrence and progression of prostate cancer, and the MACC1, c-Met, Apaf-1 and Caspase-9 expression in prostate cancer tissue are regulated by miRNAs.

  18. Construction of shRNA lentiviral vector

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    Hong Song

    2010-12-01

    Full Text Available Lentiviruses have been adapted as gene delivery vehicles. This article summarized shRNA lentiviral vector methods generally used in research laboratories. The main procedures of shRNA lentiviral vector include that (1 Target sequences screening and shRNA oligonucleotides designing, (2 insert designed oligonucleotides into lentiviral vectors, (3 using packaging cells to produce shRNA lentivirus, and (4 transducing target cells with shRNA lentivirus.

  19. Energy profile and secondary structure impact shRNA efficacy

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    Zeng Xiao

    2009-07-01

    Full Text Available Abstract Background RNA interference (RNAi is a cellular mechanism in which a short/small double stranded RNA induces the degradation of its sequence specific target mRNA, leading to specific gene silencing. Since its discovery, RNAi has become a powerful biological technique for gene function studies and drug discovery. The very first requirement of applying RNAi is to design functional small interfering RNA (siRNA that can uniquely induce the degradation of the targeted mRNA. It has been shown that many functional synthetic siRNAs share some common characteristics, such as GC content limitation and free energy preferences at both terminals, etc. Results Our three-phase algorithm was developed to design siRNA on a whole-genome scale based on those identified characteristics of functional siRNA. When this algorithm was applied to design short hairpin RNA (shRNA, the validated success rate of shRNAs was over 70%, which was almost double the rate reported for TRC library. This indicates that the designs of siRNA and shRNA may share the same concerns. Further analysis of the shRNA dataset of 444 designs reveals that the high free energy states of the two terminals have the largest positive impact on the shRNA efficacy. Enforcing these energy characteristics of both terminals can further improve the shRNA design success rate to 83.1%. We also found that functional shRNAs have less probability for their 3' terminals to be involved in mRNA secondary structure formation. Conclusion Functional shRNAs prefer high free energy states at both terminals. High free energy states of the two terminals were found to be the largest positive impact factor on shRNA efficacy. In addition, the accessibility of the 3' terminal is another key factor to shRNA efficacy.

  20. Gene Electrotransfer of Plasmid with Tissue Specific Promoter Encoding shRNA against Endoglin Exerts Antitumor Efficacy against Murine TS/A Tumors by Vascular Targeted Effects.

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    Monika Stimac

    Full Text Available Vascular targeted therapies, targeting specific endothelial cell markers, are promising approaches for the treatment of cancer. One of the targets is endoglin, transforming growth factor-β (TGF-β co-receptor, which mediates proliferation, differentiation and migration of endothelial cells forming neovasculature. However, its specific, safe and long-lasting targeting remains the challenge. Therefore, in our study we evaluated the transfection efficacy, vascular targeted effects and therapeutic potential of the plasmid silencing endoglin with the tissue specific promoter, specific for endothelial cells marker endothelin-1 (ET (TS plasmid, in comparison to the plasmid with constitutive promoter (CON plasmid, in vitro and in vivo. Tissue specificity of TS plasmid was demonstrated in vitro on several cell lines, and its antiangiogenic efficacy was demonstrated by reducing tube formation of 2H11 endothelial cells. In vivo, on a murine mammary TS/A tumor model, we demonstrated good antitumor effect of gene electrotransfer (GET of either of both plasmids in treatment of smaller tumors still in avascular phase of growth, as well as on bigger tumors, already well vascularized. In support to the observations on predominantly vascular targeted effects of endoglin, histological analysis has demonstrated an increase in necrosis and a decrease in the number of blood vessels in therapeutic groups. A significant antitumor effect was observed in tumors in avascular and vascular phase of growth, possibly due to both, the antiangiogenic and the vascular disrupting effect. Furthermore, the study indicates on the potential use of TS plasmid in cancer gene therapy since the same efficacy as of CON plasmid was determined.

  1. Pooled shRNA screenings: computational analysis.

    Science.gov (United States)

    Yu, Jiyang; Putcha, Preeti; Califano, Andrea; Silva, Jose M

    2013-01-01

    Genome-wide RNA interference screening has emerged as a powerful tool for functional genomic studies of disease-related phenotypes and the discovery of molecular therapeutic targets for human diseases. Commercial short hairpin RNA (shRNA) libraries are commonly used in this area, and state-of-the-art technologies including microarray and next-generation sequencing have emerged as powerful methods to analyze shRNA-triggered phenotypes. However, computational analysis of this complex data remains challenging due to noise and small sample size from such large-scaled experiments. In this chapter we discuss the pipelines and statistical methods of processing, quality assessment, and post-analysis for both microarray- and sequencing-based screening data.

  2. Construction of lentiviral vector of shRNA specific for rat PAX2 and the silencing effect%大鼠PAX2基因RNA干扰慢病毒载体的构建及沉默效果

    Institute of Scientific and Technical Information of China (English)

    李里; 吴玉斌

    2014-01-01

    Objective To observe the influence of lentiviral vector-mediated RNA interference on expression of rat PAX 2 gene in rat normal renal tubular epithelial cells (NRK52E).Methods The effective RNA interference sequence targeting PAX 2 gene was screened and determined according to previous study.The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into pGCSIL-GFP vector to construct a lentiviral vector which expressed PAX 2 shRNA,subsequently confirmed by PCR and DNA sequencing analysis.293 T cells were co-transfected with the plasmids using liposome transfection methods ,and packaged to produce lentiviral particles.The recombinant lentiviruses were transfected into NRK52 E cells, and the PAX2 mRNA and protein expression were examined by RT-PCR and Western blot .The results were compared with those of the non-transfected and blank vector trans-fected NRK52E cells.Results PCR analysis and DNA sequencing confirmed that the PAX 2 shRNA sequences were successfully inserted in-to the lentiviral vectors.After transfection with PAX2 shRNA,the PAX2 expression in NRK52E cells was significantly inhibited at both mR-NA and protein levels compared with that in non-transfected and empty vector transfected NRK 52E cells.Conclusions The lentiviral RNAi vectors targeting PAX2 gene have been successfully constructed ,and can effectively inhibit the expression of PAX 2 gene in NRK52E cells.It paves a way for PAX2-targeted gene therapy of obstructive nephropathy.%目的:构建PAX2特异性shRNA 干扰慢病毒载体并观察其对大鼠肾小管上皮细胞( NRK52E)中PAX2基因的沉默效果。方法采用PAX2基因 RNAi靶点序列合成靶序列的 Oligo DNA,退火形成双链 DNA,与pGCSIL-GFP载体连接产生 shRNA慢病毒载体,经 PCR筛选阳性克隆进行 DNA 测序鉴定。用脂质体转染法将质粒共转染293T 细胞,将包装产生的慢病毒颗粒感染NRK52E

  3. SNCG shRNA suppressed breast cancer cell xenograft formation and growth in nude mice

    Institute of Scientific and Technical Information of China (English)

    SHEN Pei-hong; FAN Qing-xia; LI Yan-wei; ZHANG Wei; HE Xiao-kai; WANG Zhen; ZHANG Yun-han

    2011-01-01

    Background Overexpression of breast cancer-specific gene 1 (SNCG) is associated with poor prognosis in advanced breast cancer patients. This study aimed to determine the effects of SNCG knockdown in breast cancer cells by using small hairpin RNA (shRNA).Methods Four different SNCG shRNA oligonucleotides were designed and chemically synthesized to construct mammalian expression vectors. These vectors were then stably transfected into a breast cancer MCF-7 cell line to knockdown SNCG expression. After SNCG knockdown was confirmed, the stable cell lines were inoculated into nude mice. SNCG mRNA and protein expressions were analyzed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively in both the stable cell lines and xenografts.Results All four SNCG shRNA constructs significantly reduced SNCG mRNA and protein levels in MCF-7 cells, as compared to the unrelated sequence control shRNA and the liposome control mice (P<0.05). SNCG-knockdown MCF-7cells formed significantly smaller tumor masses than cells expressing the unrelated sequence control or the liposome control mice (P<0.05).Conclusion SNCG shRNA effectively suppressed breast cancer cell formation in vivo and may be a useful clinical strategy to control breast cancer.

  4. RNAi Codex: a portal/database for short-hairpin RNA (shRNA) gene-silencing constructs.

    Science.gov (United States)

    Olson, A; Sheth, N; Lee, J S; Hannon, G; Sachidanandam, R

    2006-01-01

    Use of RNA interference (RNAi) in forward genetic screens is proliferating. Currently, short-interfering RNAs (siRNAs) and short-hairpin RNAs (shRNAs) are being used to silence genes to tease out functional information. It is becoming easier to harness RNAi to silence specific genes, owing to the development of libraries of readymade shRNA and siRNA gene-silencing constructs by using a variety of sources. RNAi Codex, which consists of a database of shRNA related information and an associated website, has been developed as a portal for publicly available shRNA resources and is accessible at http://codex.cshl.org. RNAi Codex currently holds data from the Hannon-Elledge shRNA library and allows the use of biologist-friendly gene names to access information on shRNA constructs that can silence the gene of interest. It is designed to hold user-contributed annotations and publications for each construct, as and when such data become available. We will describe features of RNAi Codex and explain the use of the tool.

  5. Is TNF-a-targeted short hairpin RNA (shRNA) a novel potential therapeutic tool in psoriasis treatment?

    DEFF Research Database (Denmark)

    Stenderup, Karin; Jakobsen, Maria; Rosada, Cecilia

    2008-01-01

      TNF-α is a well known target in psoriasis treatment and biological treatments targeting TNF-a are already clinically used against psoriasis and psoriasis arthritis. Attention is however given to a novel therapeutic tool: RNA interference that controls gene silencing. This study investigates...... the efficiency of targeting TNF-a with specific short hairpin RNA (shRNA) and explores its potential in treating psoriasis. ShRNAs targeting human TNF-α mRNA were generated. Their efficiency in down-regulating TNF-a protein expression was evaluated using a Renilla luciferase screening-assay and a transient co......-transfection assay. The expression cassette encoding the most efficient TNF-a shRNA was inserted into a lentiviral vector, allowing proficient gene delivery. The lentiviral vector is integrated into the host genome and establishes life-long infection and persistent shRNA expression. The lentiviral vector expressing...

  6. Expression of plasmid-based shRNA against the E1 and nsP1 genes effectively silenced Chikungunya virus replication.

    Directory of Open Access Journals (Sweden)

    Shirley Lam

    Full Text Available BACKGROUND: Chikungunya virus (CHIKV is a re-emerging alphavirus that causes chikungunya fever and persistent arthralgia in humans. Currently, there is no effective vaccine or antiviral against CHIKV infection. Therefore, this study evaluates whether RNA interference which targets at viral genomic level may be a novel antiviral strategy to inhibit the medically important CHIKV infection. METHODS: Plasmid-based small hairpin RNA (shRNA was investigated for its efficacy in inhibiting CHIKV replication. Three shRNAs designed against CHIKV Capsid, E1 and nsP1 genes were transfected to establish stable shRNA-expressing cell clones. Following infection of stable shRNA cells clones with CHIKV at M.O.I. 1, viral plaque assay, Western blotting and transmission electron microscopy were performed. The in vivo efficacy of shRNA against CHIKV replication was also evaluated in a suckling murine model of CHIKV infection. RESULTS: Cell clones expressing shRNAs against CHIKV E1 and nsP1 genes displayed significant inhibition of infectious CHIKV production, while shRNA Capsid demonstrated a modest inhibitory effect as compared to scrambled shRNA cell clones and non-transfected cell controls. Western blot analysis of CHIKV E2 protein expression and transmission electron microscopy of shRNA E1 and nsP1 cell clones collectively demonstrated similar inhibitory trends against CHIKV replication. shRNA E1 showed non cell-type specific anti-CHIKV effects and broad-spectrum silencing against different geographical strains of CHIKV. Furthermore, shRNA E1 clones did not exert any inhibition against Dengue virus and Sindbis virus replication, thus indicating the high specificity of shRNA against CHIKV replication. Moreover, no shRNA-resistant CHIKV mutant was generated after 50 passages of CHIKV in the stable cell clones. More importantly, strong and sustained anti-CHIKV protection was conferred in suckling mice pre-treated with shRNA E1. CONCLUSION: Taken together, these

  7. Combination of Endothelial-Monocyte-Activating Polypeptide-II with Temozolomide Suppress Malignant Biological Behaviors of Human Glioblastoma Stem Cells via miR-590-3p/MACC1 Inhibiting PI3K/AKT/mTOR Signal Pathway

    Science.gov (United States)

    Zhou, Wei; Liu, Libo; Xue, Yixue; Zheng, Jian; Liu, Xiaobai; Ma, Jun; Li, Zhen; Liu, Yunhui

    2017-01-01

    This study aims to investigate the effect of Endothelial-Monocyte-Activating Polypeptide-II (EMAP-II) combined with temozolomide (TMZ) upon glioblastoma stem cells (GSCs) and its possible molecular mechanisms. In this study, combination of EMAP-II with TMZ inhibited cell viability, migration and invasion in GSCs, and autophagy inhibitor 3-methyl adenine (3-MA) and chloroquine (CQ) partly reverse the anti-proliferative effect of the combination treatment. Autophagic vacuoles were formed in GSCs after the combination therapy, accompanied with the up-regulation of LC3-II and Beclin-1 as well as the down-regulation of p62/SQSTM1. Further, miR-590-3p was up-regulated and Metastasis-associated in colon cancer 1 (MACC1) was down-regulated by the combination treatment in GSCs; MiR-590-3p overexpression and MACC1 knockdown up-regulated LC3-II and Beclin-1 as well as down-regulated p62/SQSTM1 in GSCs; MACC1 was identified as a direct target of miR-590-3p, mediating the effects of miR-590-3p in the combination treatment. Furthermore, the combination treatment and MACC1 knockdown decreased p-PI3K, p-Akt, p-mTOR, p-S6 and p-4EBP in GSCs; PI3K/Akt agonist insulin-like growth factor-1(IGF-1) partly blocked the effect of the combination treatment. Moreover, in vivo xenograft models, the mice given stable overexpressed miR-590-3p cells and treated with EMAP-II and TMZ had the smallest tumor sizes, besides, miR-590-3p + EMAP-II + TMZ up-regulated the expression level of miR-590-3p, LC3-II and Beclin-1 as well as down-regulated p62/SQSTM1. In conclusion, these results elucidated anovel molecular mechanism of EMAP-II in combination with TMZ suppressed malignant biological behaviors of GSCs via miR-590-3p/MACC1 inhibiting PI3K/AKT/mTOR signaling pathway, and might provide potential therapeutic approaches for human GSCs.

  8. Plasmid-based Survivin shRNA and GRIM-19 carried by attenuated Salmonella suppresses tumor cell growth

    Institute of Scientific and Technical Information of China (English)

    Yan-Bo Liu1; De-Qi Xu; Ling Zhang1; Ya-Xiong Guo; Li-Fang Gao; Xi-Chun Liu; Li-Juan Zhao; Bao-Feng Guo; Li-Jing Zhao; Xue-Jian Zhao

    2012-01-01

    Persistent activation of Survivin and its overexpression contribute to the formation,progression and metastasis of several different tumor types.Therefore,Survivin is an ideal target for RNA interference mediated-growth inhibition.Blockade of Survivin using specific short hairpin RNAs (shRNA) can significantly reduce prostate tumor growth.RNA interference does not fully ablate target gene expression,owing to the idiosyncrasies associated with shRNAs and their targets.To enhance the therapeutic efficacy of Survivin-specific shRNA,we employed a combinatorial expression of Survivin-specific shRNA and gene associated with retinoid-interferon-induced mortality-19 (GRIM-19).Then,the GRIM-19 coding sequences and Survivin-specific shRNAs were used to create a dual expression plasmid vector and were carried by an attenuated strain of Salmonella enteric serovar typhimurium (S.typhimurium) to treat prostate cancer in vitroand in vivo.We found that the co-expressed Survivin-specific shRNA and GRIM-19synergistically and more effectively inhibited prostate tumor proliferation and survival,when compared with treatment with either single agent alone in vitro and in vivo.This study has provided a novel cancer gene therapeutic approach for prostate cancer.

  9. Construction and Characterization of Lentiviral shRNA Expression Vector Targeting Rat CD40 Gene in Dendritic Cells

    Institute of Scientific and Technical Information of China (English)

    SUN Mei; LI Jin-dong; JIANG Rui; JIN Cheng-yan; GAO Nan; LUO Shu-li; WANG Chun-guang; WANG Bin; WANG Rong-you; ZHANG Xing-yi

    2009-01-01

    To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells(DCs), specific siRNA targets with short hairpin frame were designed and synthesized according to the mRNA sequence of rat CD40 gene. DNA oligo was cloned into lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. The verified plasmids were transfected into 293T cells that over-express recombinant CD40 in order to select the most effective siRNA targets. shRNA lentivi-ruses from the selected constructs were propagated and harvested with a virus packaging system, and the virus titers were determined. Western blot and Real-time PCR were performed to determine CD40 expression level in the virus-infected dendritic cells. PCR and sequencing analyses reveal that shRNA plasmids of four targets were successfully constructed. The optimal interfering target was selected, and the virus with a titer of 5×10~7 TU/mL was successfully packaged. CD40 expression in rat DCs was knockdown at both mRNA and protein levels by virus infection. In comparison to that of control groups, CD40 mRNA expression and protein expression were decreased by 60.9% and 61.2%, respectively. We have successfully constructed recombinant lentiviral shRNA expression vector targeting rat CD40 gene that can effectively down-regulate CD40 gene expression at mRNA and protein levels in rat DC.

  10. An RNAi in silico approach to find an optimal shRNA cocktail against HIV-1.

    Science.gov (United States)

    Méndez-Ortega, María C; Restrepo, Silvia; Rodríguez-R, Luis M; Pérez, Iván; Mendoza, Juan C; Martínez, Andrés P; Sierra, Roberto; Rey-Benito, Gloria J

    2010-12-20

    HIV-1 can be inhibited by RNA interference in vitro through the expression of short hairpin RNAs (shRNAs) that target conserved genome sequences. In silico shRNA design for HIV has lacked a detailed study of virus variability constituting a possible breaking point in a clinical setting. We designed shRNAs against HIV-1 considering the variability observed in naïve and drug-resistant isolates available at public databases. A Bioperl-based algorithm was developed to automatically scan multiple sequence alignments of HIV, while evaluating the possibility of identifying dominant and subdominant viral variants that could be used as efficient silencing molecules. Student t-test and Bonferroni Dunn correction test were used to assess statistical significance of our findings. Our in silico approach identified the most common viral variants within highly conserved genome regions, with a calculated free energy of ≥ -6.6 kcal/mol. This is crucial for strand loading to RISC complex and for a predicted silencing efficiency score, which could be used in combination for achieving over 90% silencing. Resistant and naïve isolate variability revealed that the most frequent shRNA per region targets a maximum of 85% of viral sequences. Adding more divergent sequences maintained this percentage. Specific sequence features that have been found to be related with higher silencing efficiency were hardly accomplished in conserved regions, even when lower entropy values correlated with better scores. We identified a conserved region among most HIV-1 genomes, which meets as many sequence features for efficient silencing. HIV-1 variability is an obstacle to achieving absolute silencing using shRNAs designed against a consensus sequence, mainly because there are many functional viral variants. Our shRNA cocktail could be truly effective at silencing dominant and subdominant naïve viral variants. Additionally, resistant isolates might be targeted under specific antiretroviral selective

  11. An RNAi in silico approach to find an optimal shRNA cocktail against HIV-1

    Directory of Open Access Journals (Sweden)

    Mendoza Juan C

    2010-12-01

    Full Text Available Abstract Background HIV-1 can be inhibited by RNA interference in vitro through the expression of short hairpin RNAs (shRNAs that target conserved genome sequences. In silico shRNA design for HIV has lacked a detailed study of virus variability constituting a possible breaking point in a clinical setting. We designed shRNAs against HIV-1 considering the variability observed in naïve and drug-resistant isolates available at public databases. Methods A Bioperl-based algorithm was developed to automatically scan multiple sequence alignments of HIV, while evaluating the possibility of identifying dominant and subdominant viral variants that could be used as efficient silencing molecules. Student t-test and Bonferroni Dunn correction test were used to assess statistical significance of our findings. Results Our in silico approach identified the most common viral variants within highly conserved genome regions, with a calculated free energy of ≥ -6.6 kcal/mol. This is crucial for strand loading to RISC complex and for a predicted silencing efficiency score, which could be used in combination for achieving over 90% silencing. Resistant and naïve isolate variability revealed that the most frequent shRNA per region targets a maximum of 85% of viral sequences. Adding more divergent sequences maintained this percentage. Specific sequence features that have been found to be related with higher silencing efficiency were hardly accomplished in conserved regions, even when lower entropy values correlated with better scores. We identified a conserved region among most HIV-1 genomes, which meets as many sequence features for efficient silencing. Conclusions HIV-1 variability is an obstacle to achieving absolute silencing using shRNAs designed against a consensus sequence, mainly because there are many functional viral variants. Our shRNA cocktail could be truly effective at silencing dominant and subdominant naïve viral variants. Additionally, resistant

  12. Silencing of Inducible Immunoproteasome Subunit Expression by Chemically Modified siRNA and shRNA.

    Science.gov (United States)

    Gvozdeva, Olga V; Prassolov, Vladimir S; Zenkova, Marina A; Vlassov, Valentin V; Chernolovskaya, Elena L

    2016-08-01

    Overexpression of inducible subunits of immunoproteasome is related to pathogenesis of some chronic diseases. Specific inhibition of the immunosubunits may be used for the treatment of these diseases and RNA interference is one of the potent methods used in this area. We designed 2'-O-methyl modified siRNAs with selectively protected nuclease-sensitive sites, which efficiently silence LMP2, LMP7, and MECL-1 genes expression. To provide stable long-lasting inhibition of target genes, short-hairpin RNAs (shRNA) expressed by lentiviral vectors were constructed. Our results demonstrated that chemically modified siRNAs inhibited the expression of target genes with similar efficiency or with efficiency exceeding that of corresponding shRNAs and provide silencing effect for 5 days.

  13. A primer on using pooled shRNA libraries for functional genomic screens

    Institute of Scientific and Technical Information of China (English)

    Guang Hu; Ji Luo

    2012-01-01

    The discovery of RNA interference (RNAi) has revolutionized genetic analysis in mammalian cells.Loss-of-function RNAi screens enable rapid,functional annotation of the genome.Of the various RNAi approaches,pooled shRNA libraries have received considerable attention because of their versatility.A number of genome-wide shRNA libraries have been constructed against the human and mouse genomes,and these libraries can be readily applied to a variety of screens to interrogate the function of human and mouse genes in an unbiased fashion.We provide an introduction to the technical aspects of using pooled shRNA libraries for genetic screens.

  14. Viral delivery of shRNA to amygdala neurons leads to neurotoxicity and deficits in Pavlovian fear conditioning.

    Science.gov (United States)

    de Solis, Christopher A; Holehonnur, Roopashri; Banerjee, Anwesha; Luong, Jonathan A; Lella, Srihari K; Ho, Anthony; Pahlavan, Bahram; Ploski, Jonathan E

    2015-10-01

    knockdown of Egr1 and GluN2A compared to the other groups examined respectively, but Arc was not knocked down in the shArc group under these conditions. Differences in fear conditioning among the shLuc, shCntrl, shArc and shEgr1 groups were not detected under these circumstances; however, the shGluN2A group exhibited significantly impaired fear conditioning compared to most of the groups, indicating that gene specific deficits in fear conditioning could be observed utilizing viral mediated delivery of shRNA. Collectively, these data indicate that viral mediated shRNA expression was toxic to neurons in vivo, under all viral titers examined and this toxicity in some cases may be masking gene specific changes in learning. Therefore, the use of this technology in behavioral neuroscience warrants a heightened level of careful consideration and potential methods to alleviate shRNA induced toxicity are discussed.

  15. Construction of a Hep-2 cell line stably transfected with Livin shRNA.

    Science.gov (United States)

    Wang, S L; Deng, W T; Wen, G F; Li, C W; Zeng, Y J

    2016-01-01

    The aim of this study was to construct a eukaryotic expression plasmid with a short hairpin RNA (shRNA) targeting Livin in order to obtain a stably transfected Hep-2 cell line with a reduced expression of Livin. The shRNA targeting Livin mRNA was designed, and a shRNA plasmid and a negative control plasmid were constructed. After amplification in E. coli, restriction endonuclease digestion and sequence confirmation, the plasmids were transfected into Hep-2 cells using Lipofectamine 2000. The stably transfected cell line was screened using G418, and inhibition of Livin mRNA and protein levels were detected using real-time PCR and western blotting, respectively. pGenesil-Livin-shRNA eukaryotic expression plasmid was successfully constructed and identified by sequencing. Green fluorescent protein (GFP) expression was observed in Hep-2 cells transfected with shRNA plasmids by fluorescence microscopy. The expression levels of Livin mRNA and protein decreased significantly in Hep-2 cells transfected with the shRNA recombinant plasmid. The mRNA level was reduced by 47.17 %, and the protein level was reduced by 34.25 %. The shRNA eukaryotic expression plasmid targeting Livin was successfully constructed, which could significantly inhibit the expression of Livin in laryngeal cancer Hep-2 cells. This provides a basis for future research on the function of Livin in Hep-2 cells, and gene therapy for laryngeal cancer.

  16. Multiplex shRNA Screening of Germ Cell Development by in Vivo Transfection of Mouse Testis

    Directory of Open Access Journals (Sweden)

    Nicholas R. Y. Ho

    2017-01-01

    Full Text Available Spermatozoa are one of the few mammalian cell types that cannot be fully derived in vitro, severely limiting the application of modern genomic techniques to study germ cell biology. The current gold standard approach of characterizing single-gene knockout mice is slow as generation of each mutant line can take 6–9 months. Here, we describe an in vivo approach to rapid functional screening of germline genes based on a new nonsurgical, nonviral in vivo transfection method to deliver nucleic acids into testicular germ cells. By coupling multiplex transfection of short hairpin RNA (shRNA constructs with pooled amplicon sequencing as a readout, we were able to screen many genes for spermatogenesis function in a quick and inexpensive experiment. We transfected nine mouse testes with a pilot pool of RNA interference (RNAi against well-characterized genes to show that this system is highly reproducible and accurate. With a false negative rate of 18% and a false positive rate of 12%, this method has similar performance as other RNAi screens in the well-described Drosophila model system. In a separate experiment, we screened 26 uncharacterized genes computationally predicted to be essential for spermatogenesis and found numerous candidates for follow-up studies. Finally, as a control experiment, we performed a long-term selection screen in neuronal N2a cells, sampling shRNA frequencies at five sequential time points. By characterizing the effect of both libraries on N2a cells, we show that our screening results from testis are tissue-specific. Our calculations indicate that the current implementation of this approach could be used to screen thousands of protein-coding genes simultaneously in a single mouse testis. The experimental protocols and analysis scripts provided will enable other groups to use this procedure to study diverse aspects of germ cell biology ranging from epigenetics to cell physiology. This approach also has great promise as an

  17. Optimization of duplex stability and terminal asymmetry for shRNA design.

    Directory of Open Access Journals (Sweden)

    Olga V Matveeva

    Full Text Available Prediction of efficient oligonucleotides for RNA interference presents a serious challenge, especially for the development of genome-wide RNAi libraries which encounter difficulties and limitations due to ambiguities in the results and the requirement for significant computational resources. Here we present a fast and practical algorithm for shRNA design based on the thermodynamic parameters. In order to identify shRNA and siRNA features universally associated with high silencing efficiency, we analyzed structure-activity relationships in thousands of individual RNAi experiments from publicly available databases (ftp://ftp.ncbi.nlm.nih.gov/pub/shabalin/siRNA/si_shRNA_selector/. Using this statistical analysis, we found free energy ranges for the terminal duplex asymmetry and for fully paired duplex stability, such that shRNAs or siRNAs falling in both ranges have a high probability of being efficient. When combined, these two parameters yield a approximately 72% success rate on shRNAs from the siRecords database, with the target RNA levels reduced to below 20% of the control. Two other parameters correlate well with silencing efficiency: the stability of target RNA and the antisense strand secondary structure. Both parameters also correlate with the short RNA duplex stability; as a consequence, adding these parameters to our prediction scheme did not substantially improve classification accuracy. To test the validity of our predictions, we designed 83 shRNAs with optimal terminal asymmetry, and experimentally verified that small shifts in duplex stability strongly affected silencing efficiency. We showed that shRNAs with short fully paired stems could be successfully selected by optimizing only two parameters: terminal duplex asymmetry and duplex stability of the hypothetical cleavage product, which also relates to the specificity of mRNA target recognition. Our approach performs at the level of the best currently utilized algorithms that take

  18. Modified gateway system for double shRNA expression and Cre/lox based gene expression

    Directory of Open Access Journals (Sweden)

    Leung Lisa

    2011-03-01

    Full Text Available Abstract Background The growing need for functional studies of genes has set the stage for the development of versatile tools for genetic manipulations. Results Aiming to provide tools for high throughput analysis of gene functions, we have developed a modified short hairpin RNA (shRNA and gene expression system based on Gateway Technology. The system contains a series of entry and destination vectors that enables easy transfer of shRNA or cDNA into lentiviral expression systems with a variety of selection or marker genes (i.e. puromycin, hygromycin, green fluorescent protein-EGFP, yellow fluorescent protein-YFP and red fluorescent protein-dsRed2. Our shRNA entry vector pENTR.hU6.hH1 containing two tandem human shRNA expression promoters, H1 and U6, was capable of co-expressing two shRNA sequences simultaneously. The entry vector for gene overexpression, pENTR.CMV.ON was constructed to contain CMV promoter with a multiple cloning site flanked by loxP sites allowing for subsequent Cre/lox recombination. Both shRNA and cDNA expression vectors also contained attL sites necessary for recombination with attR sites in our destination expression vectors. As proof of principle we demonstrate the functionality and efficiency of this system by testing expression of several cDNA and shRNA sequences in a number of cell lines. Conclusion Our system is a valuable addition to already existing library of Gateway based vectors and can be an essential tool for many aspects of gene functional studies.

  19. Efficient and nontoxic biological response carrier delivering TNF-α shRNA for gene silencing in a murine model of rheumatoid arthritis

    Directory of Open Access Journals (Sweden)

    Jialin Song

    2016-08-01

    Full Text Available Small interfering RNA (siRNA is an effective and specific method for silencing genes. However, an efficient and nontoxic carrier is needed to deliver the siRNA into the target cells. Tumor necrosis factor α (TNF-α plays a central role in the occurrence and progression of rheumatoid arthritis. In this study, we pre-synthetized a degradable cationic polymer (PDAPEI from 2,6-pyridinedicarboxaldehyde and low molecular weight polyethyleneimine (PEI, Mw=1.8 kDa as a gene vector for the delivery of TNF-α shRNA. The PDAPEI/pDNA complex showed a suitable particle size and stable zeta potential for transfection. In vitro study of the PDAPEI/pDNA complex revealed a lower cytotoxicity and higher transfection efficiency when transfecting TNF-α shRNA to macrophages by significantly down-regulating the expression of TNF-α. Moreover, the complex was extremely efficient in decreasing the severity of arthritis in mice with collagen-induced arthritis (CIA. PDAPEI delivered TNF-α shRNA has great potential in the treatment of rheumatoid arthritis.

  20. A kinase shRNA screen links LATS2 and the pRB tumor suppressor.

    Science.gov (United States)

    Tschöp, Katrin; Conery, Andrew R; Litovchick, Larisa; Decaprio, James A; Settleman, Jeffrey; Harlow, Ed; Dyson, Nicholas

    2011-04-15

    pRB-mediated inhibition of cell proliferation is a complex process that depends on the action of many proteins. However, little is known about the specific pathways that cooperate with the Retinoblastoma protein (pRB) and the variables that influence pRB's ability to arrest tumor cells. Here we describe two shRNA screens that identify kinases that are important for pRB to suppress cell proliferation and pRB-mediated induction of senescence markers. The results reveal an unexpected effect of LATS2, a component of the Hippo pathway, on pRB-induced phenotypes. Partial knockdown of LATS2 strongly suppresses some pRB-induced senescence markers. Further analysis shows that LATS2 cooperates with pRB to promote the silencing of E2F target genes, and that reduced levels of LATS2 lead to defects in the assembly of DREAM (DP, RB [retinoblastoma], E2F, and MuvB) repressor complexes at E2F-regulated promoters. Kinase assays show that LATS2 can phosphorylate DYRK1A, and that it enhances the ability of DYRK1A to phosphorylate the DREAM subunit LIN52. Intriguingly, the LATS2 locus is physically linked with RB1 on 13q, and this region frequently displays loss of heterozygosity in human cancers. Our results reveal a functional connection between the pRB and Hippo tumor suppressor pathways, and suggest that low levels of LATS2 may undermine the ability of pRB to induce a permanent cell cycle arrest in tumor cells.

  1. MISSION LentiPlex pooled shRNA library screening in mammalian cells.

    Science.gov (United States)

    Coussens, Matthew J; Corman, Courtney; Fischer, Ashley L; Sago, Jack; Swarthout, John

    2011-12-21

    RNA interference (RNAi) is an intrinsic cellular mechanism for the regulation of gene expression. Harnessing the innate power of this system enables us to knockdown gene expression levels in loss of gene function studies. There are two main methods for performing RNAi. The first is the use of small interfering RNAs (siRNAs) that are chemically synthesized, and the second utilizes short-hairpin RNAs (shRNAs) encoded within plasmids. The latter can be transfected into cells directly or packaged into replication incompetent lentiviral particles. The main advantages of using lentiviral shRNAs is the ease of introduction into a wide variety of cell types, their ability to stably integrate into the genome for long term gene knockdown and selection, and their efficacy in conducting high-throughput loss of function screens. To facilitate this we have created the LentiPlex pooled shRNA library. The MISSION LentiPlex Human shRNA Pooled Library is a genome-wide lentiviral pool produced using a proprietary process. The library consists of over 75,000 shRNA constructs from the TRC collection targeting 15,000+ human genes. Each library is tested for shRNA representation before product release to ensure robust library coverage. The library is provided in a ready-to-use lentiviral format at titers of at least 5 x 10(8) TU/ml via p24 assay and is pre-divided into ten subpools of approximately 8,000 shRNA constructs each. Amplification and sequencing primers are also provided for downstream target identification. Previous studies established a synergistic antitumor activity of TRAIL when combined with Paclitaxel in A549 cells, a human lung carcinoma cell line. In this study we demonstrate the application of a pooled LentiPlex shRNA library to rapidly conduct a positive selection screen for genes involved in the cytotoxicity of A549 cells when exposed to TRAIL and Paclitaxel. One barrier often encountered with high-throughput screens is the cost and difficulty in deconvolution; we

  2. Adenovirus-mediated shRNA interference against HSV-1 replication in vitro.

    Science.gov (United States)

    Song, Bo; Liu, Xinjing; Wang, Qingzhi; Zhang, Rui; Yang, Ting; Han, Zhiqiang; Xu, Yuming

    2016-12-01

    The UL29 and UL28 proteins encoded by herpes simplex virus type 1 (HSV-1) are critical for its replication and packaging, respectively. Research has demonstrated that synthesized siRNA molecules targeting the UL29 gene are able to suppress HSV-2 replication and the UL28-null HSV-1 gene cannot form infectious viruses in vitro. Silencing the UL28 and UL29 genes by RNAi might lead to the development of novel antiviral agents for the treatment of HSV-1 infections. Two kinds of short hairpin RNAs (shRNAs) targeting the UL29 and UL28 genes were chemically synthesized and then delivered into cells by a replication-defective human adenovirus type 5 (Adv5) vector. (-) shRNAs targeting none of the genome of HSV-1 were used as the control. Vero cells were inoculated with Ad-UL28shRNA or Ad-UL29shRNA at a multiplicity of infection (MOI) of 100 and challenged 24 h later with HSV-1 at an MOI of 0.01 to inhibit HSV-1 replication, as measured by the level of the corresponding RNA and proteins. In addition, the amount of progeny virus was assessed at daily intervals. The antiviral effects of Ad-shRNAs at ongoing HSV-1 infection were explored at 12 h after inoculation of the HSV-1. The results showed that the shRNAs delivered by Adv5 significantly suppressed HSV-1 replication in vitro, as determined by the levels of viral RNA transcription, viral protein synthesis, and viral production. The Ad-UL28shRNA and Ad-UL29shRNA suppressed the replication of HSV-1, respectively, compared with the control group (P HSV-1 infection (P HSV-1 infection.

  3. A simple and robust vector-based shRNA expression system used for RNA interference.

    Directory of Open Access Journals (Sweden)

    Xue-jun Wang

    Full Text Available BACKGROUND: RNA interference (RNAi mediated by small interfering RNAs (siRNAs or short hairpin RNAs (shRNAs has become a powerful genetic tool for conducting functional studies. Previously, vector-based shRNA-expression strategies capable of inducing RNAi in viable cells have been developed, however, these vector systems have some disadvantages, either because they were error-prone or cost prohibitive. RESULTS: In this report we described the development of a simple, robust shRNA expression system utilizing 1 long oligonucleotide or 2 short oligonucleotides for half the cost of conventional shRNA construction methods and with a >95% cloning success rate. The shRNA loop sequence and stem structure were also compared and carefully selected for better RNAi efficiency. Furthermore, an easier strategy was developed based on isocaudomers which permit rapid combination of the most efficient promoter-shRNA cassettes. Finally, using this method, the conservative target sites for hepatitis B virus (HBV knockdown were systemically screened and HBV antigen expression shown to be successfully suppressed in the presence of connected multiple shRNAs both in vitro and in vivo. CONCLUSION: This novel design describes an inexpensive and effective way to clone and express single or multiple shRNAs from the same vector with the capacity for potent and effective silencing of target genes.

  4. Enhanced salt sensitivity following shRNA silencing of neuronal TRPV1 in rat spinal cord

    Institute of Scientific and Technical Information of China (English)

    Shuang-quan YU; Donna H WANG

    2011-01-01

    Aim: To investigate the effects of selective knockdown of TRPV1 channels in the lower thoracic and upper lumbar segments of spinal cord, dorsal root ganglia (DRG) and me senteric arteries on rat blood pressure responses to high salt intake.Methods: TRPV1 short-hairpin RNA (shRNA) was delivered using intrathecal injection (6 μg.kg1-d-1, for 3 d). Levels of TRPV1 and tyrosine hydroxylase expression were determined by Western blot analysis. Systolic blood pressure and mean arterial pressure (MAP) were examined using tail-cuff and direct arterial measurement, respectively.Results: In rats injected with control shRNA, high-salt diet (HS) caused higher systolic blood pressure compared with normal-salt diet(NS) (HS:149±4 mmHg; NS:126±2 mmHg, P<0.05). Intrathecal injection of TRPV1 shRNA significantly increased the systolic blood pressure in both HS rats and NS rats (HS:169±3 mmHg; NS:139±2 mmHg). The increases was greater in HS rats than in NS rats (HS:13.9%±1.8%; NS: 9.8±0.7, P<0.05). After TRPV1 shRNA treatment, TRPV1 expression in the dorsal horn and DRG of T8-L3 segments and in mesenteric arteries was knocked down to a greater extent in HS rats compared with NS rats. Blockade of α1-adrenoceptors abolished the TRPV1 shRNA-induced pressor effects. In rats injected with TRPV1 shRNA, level of tyrosine hydroxylase in mesenteric arteries was increased to a greater extent in HS rats compared with NS rats.Conclusion: Selective knockdown of TRPV1 expression in the lower thoracic and upper lumbar segments of spinal cord, DRG, and mesenteric arteries enhanced the prohypertensive effects of high salt intake, suggesting that TRPV1 channels in these sites protect against increased salt sensitivity, possibly via suppression of sympatho-excitatory responses.

  5. Construction of shRNA of Fulminant Hepatitis Related Gene mfgl2 and Investigation of Its Biological Effects in vitro

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    This study was designed to explore the RNA interference technique in inhibition of the expression of the mouse fibrinogen like protein 2 (mfgl2), which has been reported to be involved in the development a variety of diseases including fulminant viral hepatitis. A plasmid named p-mfgl2shRNA,complementary to the sequence of mfgl2 was constructed, while another short hairpin RNA (shRNA)which was a mutated form of the mfgl2shRNA sequences was used as a control. A plasmid named pEGFP-mfgl2 expressing the mfgl2-EGFP fusion protein was also constructed for the screening of the effect of p-mfgl2shRNA on mfgl2 expression. By cotransfection of p-mfgl2shRNA and pEGFP-mfgl2 or pcDNA3.1-mfgl2 expression construct into CHO cells or HeLa cells, the inhibition of mfgl2 expression by mfgl2shRNA was analyzed by direct observation through fluorescent microscopy, FACS, RT-PCR and immunohistochemistry staining. The experiments showed the significant inhibitory effect of p-mfgl2shRNA on mfgl2 expression at 48h post-transfection in both CHO and Hela cell lines with the inhibitory efficiency as high as 80.1%. The study demonstrated that the construct of p-mfgl2shRNA successfully interfered with the mfgl2 expression in vitro.

  6. A method to assess target gene involvement in angiogenesis in vitro and in vivo using lentiviral vectors expressing shRNA.

    Directory of Open Access Journals (Sweden)

    Wayne Blosser

    Full Text Available Current methods to study angiogenesis in cancer growth and development can be difficult and costly, requiring extensive use of in vivo methodologies. Here, we utilized an in vitro adipocyte derived stem cell and endothelial colony forming cell (ADSC/ECFC co-culture system to investigate the effect of lentiviral-driven shRNA knockdown of target genes compared to a non-targeting shRNA control on cord formation using High Content Imaging. Cord formation was significantly reduced following knockdown of the VEGF receptor VEGFR2 in VEGF-driven cord formation and the FGF receptor FGFR1 in basic FGF (bFGF-driven cord formation. In addition, cord formation was significantly reduced following knockdown of the transcription factor forkhead box protein O1 (FOXO1, a protein with known positive effects on angiogenesis and blood vessel stabilization in VEGF- and bFGF-driven cord formation. Lentiviral shRNA also demonstrated utility for stable knockdown of VEGFR2 and FOXO1 in ECFCs, allowing for interrogation of protein knockdown effects on in vivo neoangiogenesis in a Matrigel plug assay. In addition to interrogating the effect of gene knockdown in endothelial cells, we utilized lentiviral shRNA to knockdown specificity protein 1 (SP1, a transcription factor involved in the expression of VEGF, in U-87 MG tumor cells to demonstrate the ability to analyze angiogenesis in vitro in a tumor-driven transwell cord formation system and in tumor angiogenesis in vivo. A significant reduction in tumor-driven cord formation, VEGF secretion, and in vivo tumor angiogenesis was observed upon SP1 knockdown. Therefore, evaluation of target gene knockdown effects in the in vitro co-culture cord formation assay in the ADSC/ECFC co-culture, ECFCs alone, and in tumor cells translated directly to in vivo results, indicating the in vitro method as a robust, cost-effective and efficient in vitro surrogate assay to investigate target gene involvement in endothelial or tumor cell

  7. Lingo-1 shRNA and Notch signaling inhibitor DAPT promote differentiation of neural stem/progenitor cells into neurons.

    Science.gov (United States)

    Wang, Jue; Ye, Zhizhong; Zheng, Shuhui; Chen, Luming; Wan, Yong; Deng, Yubin; Yang, Ruirui

    2016-03-01

    Determination of the exogenous factors that regulate differentiation of neural stem/progenitor cells into neurons, oligodendrocytes and astrocytes is an important step in the clinical therapy of spinal cord injury (SCI). The Notch pathway inhibits the differentiation of neural stem/progenitor cells and Lingo-1 is a strong negative regulator for myelination and axon growth. While Lingo-1 shRNA and N-[N-(3, 5-difluorophenacetyl)-1-alanyl]-S-Phenylglycinet-butylester (DAPT), a Notch pathway inhibitor, have been used separately to help repair SCI, the results have been unsatisfactory. Here we investigated and elucidated the preliminary mechanism for the effect of Lingo-1 shRNA and DAPT on neural stem/progenitor cells differentiation. We found that neural stem/progenitor cells from E14 rat embryos expressed Nestin, Sox-2 and Lingo-1, and we optimized the transduction of neural stem/progenitor cells using lentiviral vectors encoding Lingo-1 shRNA. The addition of DAPT decreased the expression of Notch intracellular domain (NICD) as well as the downstream genes Hes1 and Hes5. Expression of NeuN, CNPase and GFAP in DAPT treated cells and expression of NeuN in Lingo-1 shRNA treated cells confirmed differentiation of neural stem/progenitor cells into neurons, oligodendrocytes and astrocytes. These results revealed that while Lingo-1 shRNA and Notch signaling inhibitor DAPT both promoted differentiation of neural stem cells into neurons, only DAPT was capable of driving neural stem/progenitor cells differentiation into oligodendrocytes and astrocytes. Since we were able to show that both Lingo-1 shRNA and DAPT could drive neural stem/progenitor cells differentiation, our data might aid the development of more effective SCI therapies using Lingo-1 shRNA and DAPT.

  8. In vivo targeted delivery of CD40 shRNA to mouse intestinal dendritic cells by oral administration of recombinant Sacchromyces cerevisiae.

    Science.gov (United States)

    Zhang, L; Zhang, T; Wang, L; Shao, S; Chen, Z; Zhang, Z

    2014-07-01

    Short hairpin RNA (shRNA)-mediated gene regulation is a commonly used technique for gene manipulation. An efficient and safe delivery system is indispensable when shRNA is delivered into living organisms for gene therapy. Previous studies have proved that DNA and protein can be delivered into dendritic cells (DCs) by non-pathogenic Saccharomyces cerevisiae without being degraded. CD40 is closely related to apoptosis of tumor cells and some immune mechanisms. In this study, we demonstrated that recombinant yeast S. cerevisiae efficiently delivered the shRNA of immune-associated gene (CD40) into mouse intestinal DCs via oral administration. Western blot analysis of isolated intestinal DCs indicated that the inhibition of CD40 gene expression reached up to 56-91%. The secretion of cytokines such as interleukin-2 (IL-2), IL-6, IL-10, IL-12, tumor necrosis factor-α and interferon-γ in intestinal DCs had varying degrees of changes. In conclusion, we found that orally administered recombinant yeast can be used as an efficient shRNA delivery system for intestinal DC-specific gene silencing and immunomodulation in vivo.

  9. Stable Delivery of CCR5-Directed shRNA into Human Primary Peripheral Blood Mononuclear Cells and Hematopoietic Stem/Progenitor Cells via a Lentiviral Vector

    Science.gov (United States)

    Shimizu, Saki; Yadav, Swati Seth; An, Dong Sung

    2016-01-01

    RNAi is a powerful tool to achieve suppression of a specific gene expression and therefore it has tremendous potential for gene therapy applications. A number of vector systems have been developed to express short-hairpin RNAs (shRNAs) to produce siRNAs within mammalian T-cells, primary hematopoietic stem/progenitor cells (HSPC), human peripheral blood mononuclear cells, and in animal model systems. Among these, vectors based on lentivirus backbones have significantly transformed our ability to transfer shRNAs into nondividing cells, such as HSPC, resulting in high transduction efficiencies. However, delivery and long-term expression of shRNAs should be carefully optimized for efficient knock down of target gene without causing cytotoxicity in mammalian cells. Here, we describe our protocols for the development of shRNA against a major HIV co-receptor/chemokine receptor CCR5 and the use of lentiviral vectors for stable shRNA delivery and expression in primary human PBMC and HSPC. PMID:26472455

  10. Multiple shRNA combinations for near-complete coverage of all HIV-1 strains

    Directory of Open Access Journals (Sweden)

    Tran Anna

    2011-01-01

    Full Text Available Abstract Background Combinatorial RNA interference (co-RNAi approaches are needed to account for viral variability in treating HIV-1 with RNAi, as single short hairpin RNAs (shRNA are rapidly rendered ineffective by resistant strains. Current work suggests that 4 simultaneously expressed shRNAs may prevent the emergence of resistant strains. Results In this study we assembled combinations of highly-conserved shRNAs to target as many HIV-1 strains as possible. We analyzed intersecting conservations of 10 shRNAs to find combinations with 4+ matching the maximum number of strains using 1220+ HIV-1 sequences from the Los Alamos National Laboratory (LANL. We built 26 combinations of 2 to 7 shRNAs with up to 87% coverage for all known strains and 100% coverage of clade B subtypes, and characterized their intrinsic suppressive activities in transient expression assays. We found that all combinations had high combined suppressive activities, though there were also large changes in the individual activities of the component shRNAs in our multiple expression cassette configurations. Conclusion By considering the intersecting conservations of shRNA combinations we have shown that it is possible to assemble combinations of 6 and 7 highly active, highly conserved shRNAs such that there is always at least 4 shRNAs within each combination covering all currently known variants of entire HIV-1 subtypes. By extension, it may be possible to combine several combinations for complete global coverage of HIV-1 variants.

  11. Ultrasound microbubbles combined with liposome-mediated pNogo-R shRNA delivery into neural stem cells

    Institute of Scientific and Technical Information of China (English)

    Weixia Ye; Xueping Huang; Yangyang Sun; Hao Liu; Jin Jiang; Youde Cao

    2012-01-01

    In the present study, ultrasound-mediated microbubble destruction (UMMD) alone and combined with liposome technology was used as a novel nonviral technique to transfect a Nogo receptor (Nogo-R) shRNA plasmid (pNogo-R shRNA) into neural stem cells (NSCs). Using green fluorescent protein as a reporter gene, transfection efficiency of NSCs was significantly higher in the group transfected with UMMD combined with liposomes compared with that of the group transfected with UMMD or liposomes alone, and did not affect cell vitality. In addition, Nogo-R mRNA and protein expression was dramatically decreased in the UMMD combined with liposome-mediated group compared with that of other groups after 24 hours of transfection. The UMMD technique combined with liposomes is a noninvasive gene transfer method, which showed minimal effects on cell viability and effectively increased transfer of Nogo-R shRNA into NSCs.

  12. Ultrasound microbubbles combined with liposome-mediated pNogo-R shRNA delivery into neural stem cells.

    Science.gov (United States)

    Ye, Weixia; Huang, Xueping; Sun, Yangyang; Liu, Hao; Jiang, Jin; Cao, Youde

    2012-01-01

    In the present study, ultrasound-mediated microbubble destruction (UMMD) alone and combined with liposome technology was used as a novel nonviral technique to transfect a Nogo receptor (Nogo-R) shRNA plasmid (pNogo-R shRNA) into neural stem cells (NSCs). Using green fluorescent protein as a reporter gene, transfection efficiency of NSCs was significantly higher in the group transfected with UMMD combined with liposomes compared with that of the group transfected with UMMD or liposomes alone, and did not affect cell vitality. In addition, Nogo-R mRNA and protein expression was dramatically decreased in the UMMD combined with liposome-mediated group compared with that of other groups after 24 hours of transfection. The UMMD technique combined with liposomes is a noninvasive gene transfer method, which showed minimal effects on cell viability and effectively increased transfer of Nogo-R shRNA into NSCs.

  13. Adenovirus vectors lacking virus-associated RNA expression enhance shRNA activity to suppress hepatitis C virus replication

    Science.gov (United States)

    Pei, Zheng; Shi, Guoli; Kondo, Saki; Ito, Masahiko; Maekawa, Aya; Suzuki, Mariko; Saito, Izumu; Suzuki, Tetsuro; Kanegae, Yumi

    2013-12-01

    First-generation adenovirus vectors (FG AdVs) expressing short-hairpin RNA (shRNA) effectively downregulate the expressions of target genes. However, this vector, in fact, expresses not only the transgene product, but also virus-associated RNAs (VA RNAs) that disturb cellular RNAi machinery. We have established a production method for VA-deleted AdVs lacking expression of VA RNAs. Here, we showed that the highest shRNA activity was obtained when the shRNA was inserted not at the popularly used E1 site, but at the E4 site. We then compared the activities of shRNAs against hepatitis C virus (HCV) expressed from VA-deleted AdVs or conventional AdVs. The VA-deleted AdVs inhibited HCV production much more efficiently. Therefore, VA-deleted AdVs were more effective than the currently used AdVs for shRNA downregulation, probably because of the lack of competition between VA RNAs and the shRNAs. These VA-deleted AdVs might enable more effective gene therapies for chronic hepatitis C.

  14. 11β-Hydroxysteroid dehydrogenase type 1 shRNA ameliorates glucocorticoid-induced insulin resistance and lipolysis in mouse abdominal adipose tissue.

    Science.gov (United States)

    Wang, Ying; Yan, Chaoying; Liu, Limei; Wang, Wei; Du, Hanze; Fan, Winnie; Lutfy, Kabirullah; Jiang, Meisheng; Friedman, Theodore C; Liu, Yanjun

    2015-01-01

    Long-term glucocorticoid exposure increases the risk for developing type 2 diabetes. Prereceptor activation of glucocorticoid availability in target tissue by 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) coupled with hexose-6-phosphate dehydrogenase (H6PDH) is an important mediator of the metabolic syndrome. We explored whether the tissue-specific modulation of 11β-HSD1 and H6PDH in adipose tissue mediates glucocorticoid-induced insulin resistance and lipolysis and analyzed the effects of 11β-HSD1 inhibition on the key lipid metabolism genes and insulin-signaling cascade. We observed that corticosterone (CORT) treatment increased expression of 11β-HSD1 and H6PDH and induced lipase HSL and ATGL with suppression of p-Thr(172) AMPK in adipose tissue of C57BL/6J mice. In contrast, CORT induced adipose insulin resistance, as reflected by a marked decrease in IR and IRS-1 gene expression with a reduction in p-Thr(308) Akt/PKB. Furthermore, 11β-HSD1 shRNA attenuated CORT-induced 11β-HSD1 and lipase expression and improved insulin sensitivity with a concomitant stimulation of pThr(308) Akt/PKB and p-Thr(172) AMPK within adipose tissue. Addition of CORT to 3T3-L1 adipocytes enhanced 11β-HSD1 and H6PDH and impaired p-Thr(308) Akt/PKB, leading to lipolysis. Knockdown of 11β-HSD1 by shRNA attenuated CORT-induced lipolysis and reversed CORT-mediated inhibition of pThr(172) AMPK, which was accompanied by a parallel improvement of insulin signaling response in these cells. These findings suggest that elevated adipose 11β-HSD1 expression may contribute to glucocorticoid-induced insulin resistance and adipolysis. Copyright © 2015 the American Physiological Society.

  15. Degradation of HER2/neu by ANT2 shRNA suppresses migration and invasiveness of breast cancer cells

    Directory of Open Access Journals (Sweden)

    Kim Chul-Woo

    2010-07-01

    Full Text Available Abstract Background In breast cancer, the HER2/neu oncoprotein, which belongs to the epidermal growth factor receptor family, may trigger activation of the phosphoinositide-3 kinase (PI3K/Akt pathway, which controls cell proliferation, survival, migration, and invasion. In this study, we examined the question of whether or not adenine nucleotide translocase 2 (ANT2 short hairpin RNA (shRNA-mediated down-regulation of HER2/neu and inhibitory effects on the PI3K/Akt signaling pathway suppressed migration and invasiveness of breast cancer cells. Methods We utilized an ANT2 vector-based RNA interference approach to inhibition of ANT2 expression, and the HER2/neu-overexpressing human breast cancer cell line, SK-BR3, was used throughout the study. Results In this study, ANT2 shRNA decreased HER2/neu protein levels by promoting degradation of HER2/neu protein through dissociation from heat shock protein 90 (HSP90. As a result, ANT2 shRNA induced inhibitory effects on the PI3K/Akt signaling pathway. Inhibition of PI3K/Akt signaling by ANT2 shRNA caused down-regulation of membrane-type 1 matrix metalloproteinase (MT1-MMP and vascular endothelial growth factor (VEGF expression, decreased matrix metalloproteinase 2 (MMP2 and MMP9 activity, and suppressed migration and invasion of breast cancer cells. Conclusions These results indicate that knock-down of ANT2 by shRNA down-regulates HER2/neu through suppression of HSP90's function and inhibits the PI3K/Akt signaling pathway, resulting ultimately in suppressed migration and invasion of breast cancer cells.

  16. Effects of shRNA Targeting Survivin on Apoptosis of Human Retinoblastoma Cell Line Hxo-rb44 in vitro

    Institute of Scientific and Technical Information of China (English)

    WANG Guojun; HU Yanhua; LI Pengcheng

    2006-01-01

    In order to construct a recombinant plasmid containing short hairpin RNA (shRNA) targeting survivin and to investigate its effect on survivin expression and cell apoptosis of human retinoblastoma cell line Hxo-rb44 in vitro, RNA interference plasmid pSIRENS that can express shRNA of survivin was designed, constructed, and transfected into human retinoblastoma cell line Hxo-rb44.Survivin and c-Myc expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Apoptosis of Hxo-rb44 cells was assayed by Honchest33258 staining and cell growth curve was drawn. The results showed that the oligonucleotide targeting survivin was identified in pSIRENS plasmid. After pSIRENS plasmid transfected, survivin and c-Myc expression in Hxo-rb44 cells was decreased significantly. Apoptotic rate of cells was up-regulated from (3.5±1.29) % to (36.1±19.66) %. The proliferation ability of Hxo-rb44 cells was inhibited. No significant effects on survivin expression and apoptosis of the cells were found when negative control plasmid was transfected. In conclusion, the plasmid containing shRNA targeting survivin was constructed successfully. It could inhibit efficiently the expression of survivin and c-Myc in human retinoblastoma cell Hxo-rb44 in vitro. The inhibition of the expression of c-Myc might be involved in the apoptosis of Hxo-rb44 cells.

  17. Silence of MCL-1 upstream signaling by shRNA abrogates multiple myeloma growth.

    Science.gov (United States)

    Wang, Mengchang; Wu, Di; Liu, Pengbo; Deng, Jiusheng

    2014-01-01

    Multiple myeloma (MM) is an incurable B-cell cancer with accumulated clonal abnormal plasma cells in bone marrow of patients. MCL-1 (myeloid cell leukemia sequence 1) protein is an anti-apoptotic molecule in MM cells and regulated by pro-inflammatory cytokine IL-6 and downstream signaling molecules STAT3, PI3K and MAPK. The purpose of this study is to investigate the effect of STAT3, PI3K and MAPK gene silence on MCL-1 expression in human MM cells and the consequence of cell survival. Lentivirus small hairpin RNA (shRNA) interference techniques were utilized to knock down STAT3, PI3K or MAPK genes. Gene and protein expression was quantified by quantitative real-time PCR and Western Blot. MM cell apoptosis was examined by annexin-V FITC/propidium iodide staining. Efficient silence of STAT3, PI3K, MAPK1 or MAPK2 gene robustly abrogated IL-6 enhanced MCL-1 expression and suppressed MM cell growth. Silencing STAT3 gene inhibited PI3K expression, silencing PI3K markedly abrogated STAT3 and MAPK production. Inhibition of MAPK2 gene by shMAPK2 suppressed STAT3, PI3K and MAPK1 expression in the cells. Silencing of STAT3, PI3K and MAPK2 together completely blocked MCL-1 expression in MM cells. There is a syngeneic effect among the three independent STAT3, PI3K and MAPK2 survival-signaling pathways related to MCL-1 expression in MM cells. shRNAs silencing of STAT3, PI3K and MAPK2 together could provide an effective strategy to treat MM.

  18. Cellular delivery of shRNA using aptamer-conjugated PLL-alkyl-PEI nanoparticles.

    Science.gov (United States)

    Askarian, Saeedeh; Abnous, Khalil; Taghavi, Sahar; Oskuee, Reza Kazemi; Ramezani, Mohammad

    2015-12-01

    Introduction of an efficient gene delivery vector is still the main challenge of gene therapy. Both polyethylenimine (PEI) and poly(l-lysine) (PLL) comprise disadvantages which limited their application. To explore whether their deficiencies could be compensated by preparing copolymers consisting of both PLL and PEI, we generated several combinations of PLL-alkyl-PEI copolymers conjugated to aptamer and evaluated their both gene delivery efficiency and down-regulation of Bcl-XL, an anti-apoptotic gene, in lung cancer cell line. PLL was conjugated to either 10% or 50% of PEI by grafting different percentages of PEI to alkylated-PLL as core. The properties of modified polymers including size, surface charge density, DNA condensation ability, buffering capacity and cytotoxicity were evaluated. According to transfection results, aptamer conjugated PLL-alkyl-10%-PEI (PLPE8%) was selected for further gene silencing study by plasmid shRNA. Decrease in Bcl-XL gene expression was estimated by both RT-PCR and western-blot experiments. The obtained results revealed that the new copolymers had appropriate nano-scale size (117-128 nm) even after aptamer conjugation (168-183 nm). Moreover, they exhibited increased transfection efficiencies by up to 1.8-5 folds and acceptable cytotoxicity. The apoptosis was induced in transfected cells by shRNA-aptamer-copolymer due to the down-regulation of mRNA and protein levels. This study suggested a new vector for targeted non-viral gene delivery with high transfection efficiency in lung cancer or pulmonary systems. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Modeling distinct osteosarcoma subtypes in vivo using Cre:lox and lineage-restricted transgenic shRNA.

    Science.gov (United States)

    Mutsaers, Anthony J; Ng, Alvin J M; Baker, Emma K; Russell, Megan R; Chalk, Alistair M; Wall, Meaghan; Liddicoat, Brain J J; Ho, Patricia W M; Slavin, John L; Goradia, Ankita; Martin, T John; Purton, Louise E; Dickins, Ross A; Walkley, Carl R

    2013-07-01

    Osteosarcoma is the most common primary cancer of bone and one that predominantly affects children and adolescents. Osteoblastic osteosarcoma represents the major subtype of this tumor, with approximately equal representation of fibroblastic and chondroblastic subtypes. We and others have previously described murine models of osteosarcoma based on osteoblast-restricted Cre:lox deletion of Trp53 (p53) and Rb1 (Rb), resulting in a phenotype most similar to fibroblastic osteosarcoma in humans. We now report a model of the most prevalent form of human osteosarcoma, the osteoblastic subtype. In contrast to other osteosarcoma models that have used Cre:lox mediated gene deletion, this model was generated through shRNA-based knockdown of p53. As is the case with the human disease the shRNA tumors most frequently present in the long bones and preferentially disseminate to the lungs; feature less consistently modeled using Cre:lox approaches. Our approach allowed direct comparison of the in vivo consequences of targeting the same genetic drivers using two different technologies, Cre:lox and shRNA. This demonstrated that the effects of Cre:lox and shRNA mediated knock-down are qualitatively different, at least in the context of osteosarcoma, and yielded distinct subtypes of osteosarcoma. Through the use of complementary genetic modification strategies we have established a model of the most common clinical subtype of osteosarcoma that was not previously represented and more fully recapitulated the clinical spectrum of this cancer. Copyright © 2013 Elsevier Inc. All rights reserved.

  20. Genetic screening of new genes responsible for cellular adaptation to hypoxia using a genome-wide shRNA library.

    Science.gov (United States)

    Yoshino, Seiko; Hara, Toshiro; Weng, Jane S; Takahashi, Yuka; Seiki, Motoharu; Sakamoto, Takeharu

    2012-01-01

    Oxygen is a vital requirement for multi-cellular organisms to generate energy and cells have developed multiple compensatory mechanisms to adapt to stressful hypoxic conditions. Such adaptive mechanisms are intricately interconnected with other signaling pathways that regulate cellular functions such as cell growth. However, our understanding of the overall system governing the cellular response to the availability of oxygen remains limited. To identify new genes involved in the response to hypoxic stress, we have performed a genome-wide gene knockdown analysis in human lung carcinoma PC8 cells using an shRNA library carried by a lentiviral vector. The knockdown analysis was performed under both normoxic and hypoxic conditions to identify shRNA sequences enriched or lost in the resulting selected cell populations. Consequently, we identified 56 candidate genes that might contribute to the cellular response to hypoxia. Subsequent individual knockdown of each gene demonstrated that 13 of these have a significant effect upon oxygen-sensitive cell growth. The identification of BCL2L1, which encodes a Bcl-2 family protein that plays a role in cell survival by preventing apoptosis, validates the successful design of our screen. The other selected genes have not previously been directly implicated in the cellular response to hypoxia. Interestingly, hypoxia did not directly enhance the expression of any of the identified genes, suggesting that we have identified a new class of genes that have been missed by conventional gene expression analyses to identify hypoxia response genes. Thus, our genetic screening method using a genome-wide shRNA library and the newly-identified genes represent useful tools to analyze the cellular systems that respond to hypoxic stress.

  1. Lentivirus-Mediated Nox4 shRNA Invasion and Angiogenesis and Enhances Radiosensitivity in Human Glioblastoma

    Directory of Open Access Journals (Sweden)

    Yongsheng Li

    2014-01-01

    Full Text Available Radioresistance remains a significant therapeutic obstacle in glioblastoma. Reactive oxygen species (ROS are associated with multiple cellular functions such as cell proliferation and apoptosis. Nox4 NADPH oxidase is abundantly expressed and has proven to be a major source of ROS production in glioblastoma. Here we investigated the effects of Nox4 on GBM tumor cell invasion, angiogenesis, and radiosensitivity. A lentiviral shRNA vector was utilized to stably knockdown Nox4 in U87MG and U251 glioblastoma cells. ROS production was measured by flow cytometry using the fluorescent probe DCFH-DA. Radiosensitivity was evaluated by clonogenic assay and survival curve was generated. Cell proliferation activity was assessed by a cell counting proliferation assay and invasion/migration potential by Matrigel invasion assay. Tube-like structure formation assay was used to evaluate angiogenesis ability in vitro and VEGF expression was assessed by MTT assay. Nox4 knockdown reduced ROS production significantly and suppressed glioblastoma cells proliferation and invasion and tumor associated angiogenesis and increased their radiosensitivity in vitro. Our results indicate that Nox4 may play a crucial role in tumor invasion, angiogenesis, and radioresistance in glioblastoma. Inhibition of Nox4 by lentivirus-mediated shRNA could be a strategy to overcome radioresistance and then improve its therapeutic efficacy for glioblastoma.

  2. shRNA library screening identifies nucleocytoplasmic transport as a mediator of BCR-ABL1 kinase-independent resistance.

    Science.gov (United States)

    Khorashad, Jamshid S; Eiring, Anna M; Mason, Clinton C; Gantz, Kevin C; Bowler, Amber D; Redwine, Hannah M; Yu, Fan; Kraft, Ira L; Pomicter, Anthony D; Reynolds, Kimberly R; Iovino, Anthony J; Zabriskie, Matthew S; Heaton, William L; Tantravahi, Srinivas K; Kauffman, Michael; Shacham, Sharon; Chenchik, Alex; Bonneau, Kyle; Ullman, Katharine S; O'Hare, Thomas; Deininger, Michael W

    2015-03-12

    The mechanisms underlying tyrosine kinase inhibitor (TKI) resistance in chronic myeloid leukemia (CML) patients lacking explanatory BCR-ABL1 kinase domain mutations are incompletely understood. To identify mechanisms of TKI resistance that are independent of BCR-ABL1 kinase activity, we introduced a lentiviral short hairpin RNA (shRNA) library targeting ∼5000 cell signaling genes into K562(R), a CML cell line with BCR-ABL1 kinase-independent TKI resistance expressing exclusively native BCR-ABL1. A customized algorithm identified genes whose shRNA-mediated knockdown markedly impaired growth of K562(R) cells compared with TKI-sensitive controls. Among the top candidates were 2 components of the nucleocytoplasmic transport complex, RAN and XPO1 (CRM1). shRNA-mediated RAN inhibition or treatment of cells with the XPO1 inhibitor, KPT-330 (Selinexor), increased the imatinib sensitivity of CML cell lines with kinase-independent TKI resistance. Inhibition of either RAN or XPO1 impaired colony formation of CD34(+) cells from newly diagnosed and TKI-resistant CML patients in the presence of imatinib, without effects on CD34(+) cells from normal cord blood or from a patient harboring the BCR-ABL1(T315I) mutant. These data implicate RAN in BCR-ABL1 kinase-independent imatinib resistance and show that shRNA library screens are useful to identify alternative pathways critical to drug resistance in CML.

  3. Tongue Epithelium Cells from shRNA Mediated Transgenic Goat Show High Resistance to Foot and Mouth Disease Virus.

    Science.gov (United States)

    Li, Wenting; Wang, Kejun; Kang, Shimeng; Deng, Shoulong; Han, Hongbing; Lian, Ling; Lian, Zhengxing

    2015-12-16

    Foot and mouth disease induced by foot and mouth disease virus (FMDV) is severe threat to cloven-hoofed domestic animals. The gene 3Dpol in FMDV genome encodes the viral RNA polymerase, a vital element for FMDV replication. In this study, a conserved 3D-7414shRNA targeting FMDV-3Dpol gene was designed and injected into pronuclear embryos to produce the transgenic goats. Sixty-one goats were produced, of which, seven goats positively integrated 3D-7414shRNA. Loss of function assay demonstrated that siRNA effectively knockdown 3Dpol gene in skin epithelium cells of transgenic goats. Subsequently, the tongue epithelium cells from transgenic and non-transgenic goats were infected with FMDV O/YS/CHA/05 strain. A significant decrease of virus titres and virus copy number was observed in cells of transgenic goats compared with that of non-transgenic goats, which indicated that 3D-7414siRNA inhibited FMDV replication by interfering FMDV-3Dpol gene. Furthermore, we found that expression of TLR7, RIG-I and TRAF6 was lower in FMDV infected cells from transgenic goats compared to that from non-transgenic goats, which might result from lower virus copy number in transgenic goats' cells. In conclusion, we successfully produced transgenic goats highly expressing 3D-7414siRNA targeting 3Dpol gene, and the tongue epithelium cells from the transgenic goats showed effective resistance to FMDV.

  4. Construction of shRNA Targeted to the Rat Angiotensin Ⅱ Type 1 Receptors and Its RNAi in Cytoplasma

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The expression vector of shRNA targeted to the rat angiotensin Ⅱ receptor gene wasconstructed and the efficacy of siRNAs to modulate the expression of target gene in the in vitro cul-tured mammalian cells was investigated for antihypertensive therapy in spontaneous hypertensive rat(SHR) at post-transcriptional level. The sense and antisense RNA oligonucleotides strands targe-ting angiotensin Ⅱ receptor mRNA were synthesized individually according to the sequence of therat angiotensin Ⅱ receptor. For preparation of duplexes, sense- and antisense-stranded oligonucle-otides were mixed and annealed, and the annealed duplexes were cloned into the pGenesil-1 vector..The rat glioma cells were transfected with constructed pGenesil-1-shRNA plasmid and scrambled plasmid. The cultured cells were collected at different phases. RT-PCR and Western blot were performed. The AT1 mRNA and protein levels behaved ultimately same. Compared to control after 48 h, AT1 mRNA levels were decreased to 35.5 %±3.0 %, and the levels reached their lowest point after 72 h (20.7 %±4 % of control). At 24 and 48 h, AT1 protein was reduced to 46.9 %±4.2 %and 36.98 % ± 3.7 % respectively compared to control and a maximum reduction was observed after 72 h of incubation (28. 1% ±4 % compared to controls). Plasmid-based shRNA expression systems targeted against the rat angiotensin Ⅱ receptor gene were generated successfully. The shRNAs with a 22-nt stem and a short loop were cleaved into small interfering dsRNA (siRNA) by the Dicer. The in vitro transcribed siRNA enables the effective silencing of gene expression to the target mRNA and leads to effective inhibition of translation of proteins and will be lay the foundation of application of gene silencing technology to hypertensive rats.

  5. Is TNF-a-targeted short hairpin RNA (shRNA) a novel potential therapeutic tool in psoriasis treatment?

    DEFF Research Database (Denmark)

    Stenderup, Karin; Jakobsen, Maria; Rosada, Cecilia

    2008-01-01

    TNF-a shRNA was used to transduce HEK293 cells and verify vector-derived TNF-a knockdown in vitro. In vivo, psoriasis skin was exposed to lentiviral TNF-a shRNAs by a single intra-dermal injection. Psoriasis skin for the in vivo study was obtained from psoriatic plaque skin biopsies that were...... transplanted onto SCID mice employing the psoriasis xenograft model. Three weeks after exposure, biopsies were taken and the epidermal thickness as an endpoint for evaluating psoriasis and the levels of TNF-a mRNA were measured. The lentiviral TNF-a shRNAs down-regulated the TNF-a expression in vitro...

  6. [Study on the effect of Klotho gene interferred by plasmid-mediated short hairpin RNA (shRNA) on sinoatrial node pacing channel gene].

    Science.gov (United States)

    Cai, Yingying; Wang, Han; Hou, Yanbin; Fang, Chenli; Tian, Peng; Wang, Guihua; Li, Lu; Deng, Juelin

    2013-06-01

    The study was aimed to assess the effect of Klotho gene and sinoatrial node pacing channel gene (HCN4 and HCN2) for studying sick sinus syndrome, with Klotho gene under the interference of Plasmid-mediated short hairpin RNA. Twenty-five C57BL/6J mice were divided into four groups, i. e, plasmid shRNA 24h group, plasmid shRNA 12h group, sodium chloride 24h group and sodium chloride 12h group. Plasmid shRNA 50microL (1microg/microL) and sodium chloride 50microl were respectively injected according to mice vena caudalis into those in plasmid shRNA group and sodium chloride group. After 12h or 24h respectively, all mice were executed and their sinoatrial node tissues were cut. The mRNA of Klotho, HCN4 and HCN2 gene were detected by RT-PCR. The results of RT-PCR showed that Klotho, HCN4 and HCN2 mRNA levels were lower compared with those in sodium chloride 12h group after 12h interference interval. The results indicated that there might be the a certain relationship between Klotho gene and sinoatrial node pacing channel gene.

  7. Co-delivery of Doxorubicin Encapsulated PLGA Nanoparticles and Bcl-xL shRNA Using Alkyl-Modified PEI into Breast Cancer Cells.

    Science.gov (United States)

    Ebrahimian, Mahboubeh; Taghavi, Sahar; Mokhtarzadeh, Ahad; Ramezani, Mohammad; Hashemi, Maryam

    2017-02-24

    In recent years, much effort has been focused on an appropriate combination of chemotherapeutic drugs and nucleic acids to exploit additive or synergistic therapeutic effects and overcome many obstacles such as the reduction of side effects and drug resistance. Short hairpin RNA (shRNA) has designed to allow the production of small interfering RNA (siRNA) within the cells and offer long-lasting silencing of target genes. In this study, alkyl-modified polyethylenimine (PEI 10 kD) was used for co-delivery of doxorubicin (DOX) encapsulated into poly lactic-co-glycolic acid (PLGA) nanoparticles (NPs) and Bcl-xL shRNA (one class of molecules that block apoptosis of tumor cells) into breast cancer cells. Our results demonstrated that modification of PEI with alkyl chain could enhance the induction of apoptosis in tumor cells by suppression of Bcl-xL gene using Bcl-xL shRNA more than PEI alone. On the other hand, DOX encapsulated into PLGA had more synergistic effect with shRNA in comparison with DOX alone. In conclusion, combination of PLGA-DOX NPs and alkyl-PEI/shRNA complexes may have promising applications in breast cancer therapy.

  8. Tissue distribution and cancer growth inhibition of magnetic lipoplex-delivered type 1 insulin-like growth factor receptor shRNA in nude mice

    Institute of Scientific and Technical Information of China (English)

    Minjian Kong; Xuebiao Li; Chunmao Wang; Chao Ding; Aiqiang Dong; Qunjun Duan; Zhonghua Shen

    2012-01-01

    The targeted delivery of therapeutic genes into specific tissues,as well as the determination of the biological fate and potential toxicity of nanoparticles,remains a highly relevant challenge for gene-based therapies.Type 1 insulin-like growth factor receptor (IGF-1R),an important oncogene,is frequently over-expressed in lung cancer and mediates cancer cell proliferation as well as tumor growth.In our previous studies,we have successfully applied gene delivery mediated by commercially available nanoparticles (CombiMAG) under a magnetic field,which suppresses IGF-1R expression in a non-small cell lung cancer cell line (A549) in vitro.In the present study,we aimed to investigate the biological distribution and target tumor suppression of magnetofection,as well as its potential toxicity via CombiMAG-carrying plasmids expressing green fluorescent protein (GFP) and short hairpin RNAs (shRNAs) targeting IGF-1R (pGFPshIGF-1Rs) in tumor-bearing mice.The peak expression in various organs appeared 48 h after transfection.Transgene expression via magnetofection was 3-fold improvement than via lipofection.On the 30th day after injection,the tumor size and weight of the CombiMAG-treated group (789.32 ± 39.43 mm3,105.5 ±6.1 mg) were significantly decreased compared with those of the lipofection group (893.83 ± 31.23 mm3,164.5 ±9.1 mg;P< 0.05),and the suppression rate was ~36%.After a 30-day observation,the injection of CombiMAG did not cause any apparent toxicity.Therefore,IGF-1R shRNA nanoparticles can be valuable and safe delivery agents for RNA interference therapy to tumors in vivo.

  9. Construction and Identification of Doxycycline-inducible shRNA Expressing Vector Targeting Porcine Nanog%猪Nanog基因四环素诱导干扰载体的构建和鉴定

    Institute of Scientific and Technical Information of China (English)

    张力; 罗一博; 牟彦双; 朱江; 李慧; 刘忠华

    2011-01-01

    , including inducible RNA polymerase Ⅲ ( polⅢ) promoters with embedded tetracycline operators (tetO) and co-expression of the tetracycline regulatory protein, TetR. In the absence of tetracycline or its derivative doxycycline, TetR homodimer binds tightly to the Tet operon ( s ) inserted within the promoter and acts to suppress transcription. Upon its addition, tetracycline binds with high affinity to the Tet repressor homodimer and causes a conformational change in the repressor that renders it unable to bind to tetO, allowing expression of the shRNA transcript. However, the process for generate stable cell line whose target gene knock-down is time-consuming multi-step process which limited transgenic animals' expansion. In addition, most shRNA expression units rely on polⅢ promoters such as the H1 or U6 promoter, but it is better to use RNA polymerase Ⅱ ( pol Ⅱ) promoters for tissue specific gene silencing. To overcome the limitation of shRNA expression systems in mammalian cells, a single vector which only incorporates tetO-hUbc promoter, tTS was used in this study. Firstly, five pGenesill. 0-shRNA plasmids were constructed and selected for the most effective shRNA target on porcine Nanog by Realtirae-PCR. In order to generate a single-vector harboring pol HI promoter allowing for rapid generation of stable cell lines for regulatable shRNA expression targets on porcine Nanog, the selected effective shRNA whose interference efficiency could reach 80% was inserted to a modified doxycycline-induced gene silencing vector TREsilencer. Realtime-PCR analysis demonstrates that efficiency of Nanog silencing could reach about 70% , with increasing concentration of doxycyline. It is a fundamental work for generating Nanog stably silenced porcine cell lines and further studying on Nanog genes* function.

  10. Combination of LC3 shRNA plasmid transfection and genistein treatment inhibited autophagy and increased apoptosis in malignant neuroblastoma in cell culture and animal models.

    Directory of Open Access Journals (Sweden)

    Nishant Mohan

    Full Text Available Malignant neuroblastoma is an extracranial solid tumor that usually occurs in children. Autophagy, which is a survival mechanism in many solid tumors including malignant neuroblastoma, deters the efficacy of conventional chemotherapeutic agents. To mimic starvation, we used 200 nM rapamycin that induced autophagy in human malignant neuroblastoma SK-N-BE2 and IMR-32 cells in cell culture and animal models. Combination of microtubule associated protein light chain 3 short hairpin RNA (LC3 shRNA plasmid transfection and genistein (GST treatment was tested for inhibiting rapamycin-induced autophagy and promoting apoptosis. The best synergistic efficacy caused the highest decrease in cell viability due to combination of 50 nM LC3 shRNA plasmid transfection and 25 µM GST treatment in rapamycin-treated SK-N-BE2 cells while combination of 100 nM LC3 shRNA plasmid transfection and 25 µM GST treatment in rapamycin-treated IMR-32 cells. Quantitation of acidic vesicular organelles confirmed that combination of LC3 shRNA plasmid transfection and GST treatment prevented rapamycin-induced autophagy due to down regulation of autophagy promoting marker molecules (LC3 II, Beclin 1, TLR-4, and Myd88 and upregulation of autophagy inhibiting marker molecules (p62 and mTOR in both cell lines. Apoptosis assays showed that combination therapy most effectively activated mitochondrial pathway of apoptosis in human malignant neuroblastoma in cell culture and animal models. Collectively, our current combination of LC3 shRNA plasmid transfection and GST treatment could serve as a promising therapeutic strategy for inhibiting autophagy and increasing apoptosis in human malignant neuroblastoma in cell culture and animal models.

  11. Combination of LC3 shRNA plasmid transfection and genistein treatment inhibited autophagy and increased apoptosis in malignant neuroblastoma in cell culture and animal models.

    Science.gov (United States)

    Mohan, Nishant; Chakrabarti, Mrinmay; Banik, Naren L; Ray, Swapan K

    2013-01-01

    Malignant neuroblastoma is an extracranial solid tumor that usually occurs in children. Autophagy, which is a survival mechanism in many solid tumors including malignant neuroblastoma, deters the efficacy of conventional chemotherapeutic agents. To mimic starvation, we used 200 nM rapamycin that induced autophagy in human malignant neuroblastoma SK-N-BE2 and IMR-32 cells in cell culture and animal models. Combination of microtubule associated protein light chain 3 short hairpin RNA (LC3 shRNA) plasmid transfection and genistein (GST) treatment was tested for inhibiting rapamycin-induced autophagy and promoting apoptosis. The best synergistic efficacy caused the highest decrease in cell viability due to combination of 50 nM LC3 shRNA plasmid transfection and 25 µM GST treatment in rapamycin-treated SK-N-BE2 cells while combination of 100 nM LC3 shRNA plasmid transfection and 25 µM GST treatment in rapamycin-treated IMR-32 cells. Quantitation of acidic vesicular organelles confirmed that combination of LC3 shRNA plasmid transfection and GST treatment prevented rapamycin-induced autophagy due to down regulation of autophagy promoting marker molecules (LC3 II, Beclin 1, TLR-4, and Myd88) and upregulation of autophagy inhibiting marker molecules (p62 and mTOR) in both cell lines. Apoptosis assays showed that combination therapy most effectively activated mitochondrial pathway of apoptosis in human malignant neuroblastoma in cell culture and animal models. Collectively, our current combination of LC3 shRNA plasmid transfection and GST treatment could serve as a promising therapeutic strategy for inhibiting autophagy and increasing apoptosis in human malignant neuroblastoma in cell culture and animal models.

  12. shRNA screening identifies JMJD1C as being required for leukemia maintenance

    DEFF Research Database (Denmark)

    Sroczynska, Patrycja; Cruickshank, V Adam; Bukowski, John-Paul

    2014-01-01

    Epigenetic regulatory mechanisms are implicated in the pathogenesis of acute myeloid and lymphoid leukemia (AML and ALL). Recent progress suggests that proteins involved in epigenetic control are amenable to drug intervention, but little is known about the cancer-specific dependency on epigenetic...

  13. PLK1shRNA and doxorubicin co-loaded thermosensitive PLGA-PEG-PLGA hydrogels for osteosarcoma treatment.

    Science.gov (United States)

    Ma, Hecheng; He, Chaoliang; Cheng, Yilong; Li, Dongsong; Gong, Yubao; Liu, Jianguo; Tian, Huayu; Chen, Xuesi

    2014-10-01

    Combination cancer therapy has emerged as crucial approach for achieving superior anti-cancer efficacy. In this study, we developed a strategy by localized co-delivery of PLK1shRNA/polylysine-modified polyethylenimine (PEI-Lys) complexes and doxorubicin (DOX) using biodegradable, thermosensitive PLGA-PEG-PLGA hydrogels for treatment of osteosarcoma. When incubated with osteosarcoma Saos-2 and MG-63 cells, the hydrogel containing PLK1shRNA/PEI-Lys and DOX displayed significant synergistic effects in promoting the apoptosis of osteosarcoma cells in vitro. After subcutaneous injection of the hydrogel containing PLK1shRNA/PEI-Lys and DOX beside the tumors of nude mice bearing osteosarcoma Saos-2 xenografts, the hydrogels exhibited superior antitumor efficacy in vivo compared to the hydrogels loaded with PLK1shRNA/PEI-Lys or DOX alone. It is noteworthy that the combination treatment in vivo led to almost complete suppression of tumor growth up to 16 days, significantly enhanced PLK1 silencing, higher apoptosis of tumor masses, as well as increased cell cycle regulation. Additionally, ex vivo histological analysis of major organs of the mice indicated that the localized treatments showed no obvious damage to the organs, suggesting lower systemic toxicity of the treatments. Therefore, the strategy of localized, sustained co-delivery of PLK1shRNA and DOX by using the biodegradable, injectable hydrogel may have potential for efficient clinical treatment of osteosarcoma. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Identification of the determinants of 2-deoxyglucose sensitivity in cancer cells by shRNA library screening.

    Science.gov (United States)

    Kobayashi, Hiroki; Nishimura, Haruna; Matsumoto, Ken; Yoshida, Minoru

    2015-11-06

    Combining glycolytic inhibition with other anti-cancer therapies is a potential approach to treating cancer. In this context, we attempted to identify genes that determine sensitivity to 2-deoxyglucose (2DG), a glycolytic inhibitor, in cancer cells using pooled shRNA libraries targeting ∼15,000 genes. The screen revealed that COPB1 and ARCN1, which are essential in retrograde transport, as determinants of sensitivity to 2DG: silencing of COPB1 or ARCN1 expression sensitized cells to 2DG toxicity. To address the mechanism of potentiation of 2DG toxicity by inhibition of COPI-mediated transport, we focused on the role of lipolysis as an alternate source of energy upon inhibition of glycolysis. In the process of lipolysis, COPI-mediated transport is required for localization to lipid droplets of adipose triglyceride lipase (ATGL), a key enzyme that produces fatty acids from triacylglycerol as a substrate for β-oxidation. The ATGL inhibitor atglistatin potentiated 2DG toxicity, consistent with a model in which a defect in COPI-mediated transport of ATGL to lipid droplets inhibits energy supply, thereby sensitizing cells to glycolytic inhibition. Collectively, our data demonstrated that a defect in COPI-mediated transport or pharmacological inhibition of ATGL potentiates 2DG toxicity in cancer cells, possibly due to a reduction in the energy supply.

  15. Depleting gene activities in early Drosophila embryos with the "maternal-Gal4-shRNA" system.

    Science.gov (United States)

    Staller, Max V; Yan, Dong; Randklev, Sakara; Bragdon, Meghan D; Wunderlich, Zeba B; Tao, Rong; Perkins, Lizabeth A; Depace, Angela H; Perrimon, Norbert

    2013-01-01

    In a developing Drosophila melanogaster embryo, mRNAs have a maternal origin, a zygotic origin, or both. During the maternal-zygotic transition, maternal products are degraded and gene expression comes under the control of the zygotic genome. To interrogate the function of mRNAs that are both maternally and zygotically expressed, it is common to examine the embryonic phenotypes derived from female germline mosaics. Recently, the development of RNAi vectors based on short hairpin RNAs (shRNAs) effective during oogenesis has provided an alternative to producing germline clones. Here, we evaluate the efficacies of: (1) maternally loaded shRNAs to knockdown zygotic transcripts and (2) maternally loaded Gal4 protein to drive zygotic shRNA expression. We show that, while Gal4-driven shRNAs in the female germline very effectively generate phenotypes for genes expressed maternally, maternally loaded shRNAs are not very effective at generating phenotypes for early zygotic genes. However, maternally loaded Gal4 protein is very efficient at generating phenotypes for zygotic genes expressed during mid-embryogenesis. We apply this powerful and simple method to unravel the embryonic functions of a number of pleiotropic genes.

  16. Anticancer activity of NOB1-targeted shRNA combination with TRAIL in epithelial ovarian cancer cells.

    Science.gov (United States)

    Lin, Yang; Xu, Tianmin; Teng, Hong; Cui, Manhua

    2015-01-01

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) based strategy is a promising targeted therapeutic approach for the treatment of ovarian cancer. However, the effectiveness of the treatment remains limited due to the inherent or acquired resistance of tumor cells to TRAIL. Our previously study demonstrated that downregulation of NOB1 (NIN1/RPN12 binding protein 1 homolog) expression by a lentiviral short hairpin RNA (shRNA) delivery system (Lv/sh-NOB1) suppressed ovarian cancer growth. Here, Lv/sh-NOB1 and TRAIL were combined and tested the effects of this combination on ovarian cancer cells to identify more effective therapeutics against ovarian cancer by several in vitro experiments. Tumor growth ability in SKVO3 xenograft nude mice was also determined to define this combination treatment effect in tumorigenesis in vivo. In vitro assay showed that Lv/sh-NOB1 in combination with TRAIL treatment in ovarian cancer cell synergistically suppressed the proliferation and colony formation, as well as induced cell apoptosis and increased the activity of caspase-3, -8 and -9. In vivo assay showed that Lv/sh-NOB1 combination with TRAIL synergistically suppressed tumor growth of nude mice model. Importantly, we found that downregulation of NOB1 could upregulate DR5 expression and active MAPK pathway, which might contribute to increase sensitivity TRAIL to ovarian cancer cells. These findings suggested that Lv/sh-NOB1 combination with TRAIL treatment may be a potential treatment approach for ovarian cancer.

  17. The promotion of functional recovery and nerve regeneration after spinal cord injury by lentiviral vectors encoding Lingo-1 shRNA delivered by Pluronic F-127.

    Science.gov (United States)

    Wu, Hong-Fu; Cen, Jing-Sheng; Zhong, Qian; Chen, Luming; Wang, Jue; Deng, David Y B; Wan, Yong

    2013-02-01

    Lingo-1 is selectively expressed on both oligodendrocytes and neurons in the central nervous system (CNS) and serves as a key negative regulator of nerve regeneration, implying a therapeutic target for spinal cord injury (SCI). Here we described a strategy to knock-down Lingo-1 expression in vivo using lentiviral vectors encoding Lingo-1 short harpin interfering RNA (shRNA) delivered by Pluronic F-127 (PF-127) gel, a non-cytotoxic scaffold and gene delivery carrier, after the complete transection of the T10 spinal cord in adult rats. We showed administration of PF-127 encapsulating Lingo-1 shRNA lentiviral vectors efficiently down-regulated the expression of Lingo-1, and exhibited transduction efficiency comparable to using vectors alone in oligodendrocyte culture in vitro. Furthermore, similar silencing effects and higher transfection efficiency were observed in vivo when Lingo-1 shRNA was co-delivered to the injured site by PF-127 gel with lower viral concentrations. Cografting of gel and Lingo-1 RNAi significantly promoted functional recovery and nerve regeneration, enhanced neurite outgrowth and synapses formation, preserved myelinated axons, and induced the proliferation of glial cells. In addition, the combined implantation also improved neuronal survival and inhibited cell apoptosis, which may be associated with the attenuation of endoplasmic reticulum (ER) stress after SCI. Together, our data indicated that delivering Lingo-1 shRNA by gel scaffold was a valuable treatment approach to SCI and PF-127 delivery of viral vectors to the spinal cord may provide strategy to study and develop therapies for SCI.

  18. Pol II-expressed shRNA knocks down Sod2 gene expression and causes phenotypes of the gene knockout in mice.

    Directory of Open Access Journals (Sweden)

    Xu-Gang Xia

    2006-01-01

    Full Text Available RNA interference (RNAi has been used increasingly for reverse genetics in invertebrates and mammalian cells, and has the potential to become an alternative to gene knockout technology in mammals. Thus far, only RNA polymerase III (Pol III-expressed short hairpin RNA (shRNA has been used to make shRNA-expressing transgenic mice. However, widespread knockdown and induction of phenotypes of gene knockout in postnatal mice have not been demonstrated. Previous studies have shown that Pol II synthesizes micro RNAs (miRNAs-the endogenous shRNAs that carry out gene silencing function. To achieve efficient gene knockdown in mammals and to generate phenotypes of gene knockout, we designed a construct in which a Pol II (ubiquitin C promoter drove the expression of an shRNA with a structure that mimics human miRNA miR-30a. Two transgenic lines showed widespread and sustained shRNA expression, and efficient knockdown of the target gene Sod2. These mice were viable but with phenotypes of SOD2 deficiency. Bigenic heterozygous mice generated by crossing these two lines showed nearly undetectable target gene expression and phenotypes consistent with the target gene knockout, including slow growth, fatty liver, dilated cardiomyopathy, and premature death. This approach opens the door of RNAi to a wide array of well-established Pol II transgenic strategies and offers a technically simpler, cheaper, and quicker alternative to gene knockout by homologous recombination for reverse genetics in mice and other mammalian species.

  19. Construction of METHFR shRNA/5-fluorouracil co-loaded folate-targeted chitosan polymeric nanoparticles and its anti-carcinoma effect on gastric cells growth

    Science.gov (United States)

    Xin, Lin; Fan, Ji-Chang; Le, Yi-Guan; Zeng, Fei; Cheng, Hua; Hu, Xiao-yun; Cao, Jia-Qing

    2016-05-01

    PEGylated and folate-targeted chitosan polymeric nanoparticles (FPNs) for the treatment of gastric carcinoma were prepared successfully. OQC-anchored folate conjugates were synthesized and used in assembling FPNs nano-system for enhancing intracellular uptake against folate receptor overexpressing cancer cells. The results indicated that folate-targeted chitosan polymeric nanoparticles (CPNs) can reverse drug-resistant SGC-7901 cells of 5-fluorouracil (5-FU) compared with non-targeted CPNs. Increased therapeutic efficiency of 5-FU/METHFR shRNA co-loaded PNs were also tested in SGC-7901 cells and compaed with 5-FU or METHFR shRNA in solution, which was associated with increased cell inhibition function for single drug group and synergistic effects of 5-FU and METHFR shRNA at 2.0 µg/mL FPNs concentration. In addition, the cell accumulation levels of 5-FU in SGC-7901 cells was time dependent for these nanoparticles. FPNs (effective diameter: 83.2 ± 1.1 nm; polydispersity index: 0.193) could significantly boost cellular accumulation of 5-FU and overcome the drug efflux mechanism of MDR than 5-FU-loaded NPNs and 5-FU in solution. In conclusion, ligand-targeted PNs can be used as a potentially effective drug delivery system.

  20. 小鼠 IL-35基因 shRNA 慢病毒载体构建与RNAi 效率的鉴定%Construction of shRNA Lentiviral Vector Targeting Mice IL-35 Gene and Identification of RNAi Efficiency

    Institute of Scientific and Technical Information of China (English)

    刘义帅; 王洪伟; 邸大琳; 付晓燕; 吴国庆; 王丽娜; 鞠吉雨

    2014-01-01

    目的:构建小鼠IL-35基因靶向shRNA干扰的慢病毒表达载体,抑制小鼠肝癌细胞Hepa1~6中IL-35的表达。方法设计合成IL-35EBI3亚基基因靶向shRNA序列,构建shRNA载体PLKO.1-IL-35 EBI3 shRNA-GFP,测序正确后,三质粒病毒包装系统(质粒载体+psPAX2+pMD2.G)包装成表达干扰IL-35 EBI3 shR-NA的慢病毒,慢病毒感染靶细胞Hepa1~6,荧光显微镜下观察感染效率,以实时定量RT-PCR分析对Hepa1~6细胞IL-35 EBI3基因表达的干扰效果。结果测序证实,成功构建了真核表达干扰载体PLKO.1-IL-35 EBI3 shRNA-GFP;并成功包装出表达干扰IL-35 EBI3 shRNA的慢病毒,以MOI值4.6pfu/细胞感染Hepa1~6细胞,镜下显示感染效率约90%;RT-PCR结果表明所构建的3个慢病毒载体PLKO.1-IL-35 EBI3 shRNA-GFP均可以有效干扰IL-35 EBI3的表达,其中E545干扰效率最高,为64%.结论成功构建IL-35EBI3亚基基因的shRNA慢病毒表达载体,该慢病毒表达载体能够在细胞水平有效沉默靶基因。%Objective To construct the shRNA lentiviral vectors targeting mice IL-35 gene and detect its effect of gene silence in Hepa1~6 cells.Methods The specific siRNA sequences targeting mice IL-35 gene were designed and cloned into eukaryotic expression vector PLKO.1-IL-35 EBI3 shRNA-GFP.After the correct sequencing identification ,the lentivirus particles targeting mice IL-35 gene were packaged with the three plasmid virus packaging system .The IL-35 gene specific shRNAs were infected into Hepa1~6 cells.Then,infection efficiency were observed by the fluorescence microscope .Real time reverse transcription PCR was performed to determine the expression level of IL-35 EBI3 mRNA.Results Sequencing results revealed that PLKO .1-IL-35 EBI3 shRNA-GFP plasmids were correctly constructed .The lentivirus particles targeting mice IL-35 gene were packaged successfully .The observed infection efficiency by the fluorescence

  1. Filaggrin silencing by shRNA directly impairs the skin barrier function of normal human epidermal keratinocytes and then induces an immune response

    Energy Technology Data Exchange (ETDEWEB)

    Dang, N.N. [Department of Dermatology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong Province (China); College of Life Science, Shandong Normal University, Jinan, Shandong Province (China); Pang, S.G. [Department of Endocrinology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong Province (China); Song, H.Y. [Department of Dermatology, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong Province (China); An, L.G. [College of Life Science, Shandong Normal University, Jinan, Shandong Province (China); Ma, X.L. [Central Laboratory, Jinan Central Hospital Affiliated to Shandong University, Jinan, Shandong Province (China)

    2014-11-14

    The objective of this study was to investigate whether a single defect in skin barrier function simulated by filaggrin silencing could induce Th2-predominant inflammation. Filaggrin gene expression was silenced in cultured normal human epidermal keratinocytes (NHEKs) using small hairpin RNA (shRNA, GTTGGCTCAAGCATATTATTT). The efficacy of silencing was confirmed by polymerase chain reaction (PCR) and Western blotting. Filaggrin-silenced cells (LV group), shRNA control cells (NC group), and noninfected cells (Blank group) were evaluated. The expression of cornified cell envelope-related proteins, including cytokeratin (CK)-5, -10, -14, loricrin, involucrin, and transglutaminase (TGM)-1, was detected by Western blotting. Interleukins (IL)-2, IL-4, IL-5, IL-12p70, IL-13, and interferon-gamma (IFN-γ) were detected by enzyme-linked immunosorbent assay (ELISA). After filaggrin was successfully silenced by shRNA, the expressions of CK-5, -10, -14, involucrin, and TGM-1 in NHEKs were significantly downregulated compared to the Blank and NC groups (P<0.05 or P<0.01); only loricrin expression was markedly upregulated (P<0.01). Filaggrin silencing also resulted in significant increases of IL-2, IL-4, IL-5, and IL-13 (P<0.05 or P<0.01), and significant decreases of IL-12p70 and IFN-γ (P<0.01) compared with cells in the Blank and NC groups. Filaggrin silencing impaired normal skin barrier function mainly by targeting the cornified cell envelope. The immune response after filaggrin silencing was characterized by Th2 cells, mainly because of the inhibition of IFN-γ expression. Lack of filaggrin may directly impair skin barrier function and then further induce the immune response.

  2. Human U6 promoter drives stronger shRNA activity than its schistosome orthologue in Schistosoma mansoni and human fibrosarcoma cells.

    Science.gov (United States)

    Duvoisin, Raphaël; Ayuk, Mary A; Rinaldi, Gabriel; Suttiprapa, Sutas; Mann, Victoria H; Lee, Clarence M; Harris, Nicola; Brindley, Paul J

    2012-06-01

    Blood flukes or schistosomes are the causative agents of human schistosomiasis, one of the major neglected tropical diseases. Draft genome sequences have been reported for schistosomes, but functional genomics tools are needed to investigate the role and essentiality of the newly reported genes. Vector based RNA interference can contribute to functional genomics analysis for schistosomes. Using mRNA encoding reporter firefly luciferase as a model target, we compared the performance of a schistosome and a human promoter from the U6 gene in driving shRNA in human fibrosarcoma cells and in cultured schistosomes. Further, both a retroviral [Murine leukemia virus (MLV)] and plasmid (piggyBac, pXL-Bac II) vector were utilized. The schistosome U6 gene promoter was 270 bp in length, the human U6 gene promoter was 264 bp; they shared 41% identity. Following transduction of both HT1080 fibrosarcoma cells and schistosomules of Schistosoma mansoni with pseudotyped MLV virions, stronger knockdown of luciferase activity was seen with the virions encoding the human U6 promoter driven shRNA than the schistosome U6 promoter. A similar trend was seen after transfection of HT1080 cells and schistosomules with the pXL-Bac-II constructs-stronger knockdown of luciferase activity was seen with constructs encoding the human compared to schistosome U6 promoter. The findings indicate that a human U6 gene promoter drives stronger shRNA activity than its schistosome orthologue, not only in a human cancer cell line but also in larval schistosomes. This RNA polymerase III promoter represents a potentially valuable component for vector based RNA interference studies in schistosomes and related platyhelminth parasites.

  3. Suppression of human breast tumors in NOD/SCID mice by CD44 shRNA gene therapy combined with doxorubicin treatment

    Directory of Open Access Journals (Sweden)

    Pham PV

    2012-05-01

    Full Text Available Phuc Van Pham1, Ngoc Bich Vu1, Thuy Thanh Duong1, Tam Thanh Nguyen1, Nhung Hai Truong1, Nhan Lu Chinh Phan1, Tue Gia Vuong1, Viet Quoc Pham1, Hoang Minh Nguyen1, Kha The Nguyen1, Nhung Thi Nguyen1, Khue Gia Nguyen1, Lam Tan Khat1, Dong Van Le2, Kiet Dinh Truong1, Ngoc Kim Phan11Laboratory of Stem Cell Research and Application, University of Science, Vietnam National University, HCM City, 2Military Medical University, Ha Noi, VietnamBackground: Breast cancer stem cells with a CD44+CD24- phenotype are the origin of breast tumors. Strong CD44 expression in this population indicates its important role in maintaining the stem cell phenotype. Previous studies show that CD44 down-regulation causes CD44+CD24- breast cancer stem cells to differentiate into non-stem cells that are sensitive to antitumor drugs and lose many characteristics of the original cells. In this study, we determined tumor suppression in non-obese severe combined immunodeficiency mice using CD44 shRNA therapy combined with doxorubicin treatment.Methods: Tumor-bearing non-obese severe combined immunodeficiency mice were established by injection of CD44+CD24- cells. To track CD44+CD24- cells, green fluorescence protein was stably transduced using a lentiviral vector prior to injection into mice. The amount of CD44 shRNA lentiviral vector used for transduction was based on CD44 down-regulation by in vitro CD44 shRNA transduction. Mice were treated with direct injection of CD44 shRNA lentiviral vector into tumors followed by doxorubicin administration after 48 hours. The effect was evaluated by changes in the size and weight of tumors compared with that of the control.Results: The combination of CD44 down-regulation and doxorubicin strongly suppressed tumor growth with significant differences in tumor sizes and weights compared with that of CD44 down-regulation or doxorubicin treatment alone. In the combination of CD44 down-regulation and doxorubicin group, the tumor weight was

  4. Intrathecal AAV serotype 9-mediated delivery of shRNA against TRPV1 attenuates thermal hyperalgesia in a mouse model of peripheral nerve injury.

    Science.gov (United States)

    Hirai, Takashi; Enomoto, Mitsuhiro; Kaburagi, Hidetoshi; Sotome, Shinichi; Yoshida-Tanaka, Kie; Ukegawa, Madoka; Kuwahara, Hiroya; Yamamoto, Mariko; Tajiri, Mio; Miyata, Haruka; Hirai, Yukihiko; Tominaga, Makoto; Shinomiya, Kenichi; Mizusawa, Hidehiro; Okawa, Atsushi; Yokota, Takanori

    2014-02-01

    Gene therapy for neuropathic pain requires efficient gene delivery to both central and peripheral nervous systems. We previously showed that an adenoassociated virus serotype 9 (AAV9) vector expressing short-hairpin RNA (shRNA) could suppress target molecule expression in the dorsal root ganglia (DRG) and spinal cord upon intrathecal injection. To evaluate the therapeutic potential of this approach, we constructed an AAV9 vector encoding shRNA against vanilloid receptor 1 (TRPV1), which is an important target gene for acute pain, but its role in chronic neuropathic pain remains unclear. We intrathecally injected it into the subarachnoid space at the upper lumbar spine of mice 3 weeks after spared nerve injury (SNI). Delivered shTRPV1 effectively suppressed mRNA and protein expression of TRPV1 in the DRG and spinal cord, and it attenuated nerve injury-induced thermal allodynia 10-28 days after treatment. Our study provides important evidence for the contribution of TRPV1 to thermal hypersensitivity in neuropathic pain and thus establishes intrathecal AAV9-mediated gene delivery as an investigative and potentially therapeutic platform for the nervous system.

  5. In vivo targeting of ADAM9 gene expression using lentivirus-delivered shRNA suppresses prostate cancer growth by regulating REG4 dependent cell cycle progression.

    Directory of Open Access Journals (Sweden)

    Che-Ming Liu

    Full Text Available Cancer cells respond to stress by activating a variety of survival signaling pathways. A disintegrin and metalloproteinase (ADAM 9 is upregulated during cancer progression and hormone therapy, functioning in part through an increase in reactive oxygen species. Here, we present in vitro and in vivo evidence that therapeutic targeting of ADAM9 gene expression by lentivirus-delivered small hairpin RNA (shRNA significantly inhibited proliferation of human prostate cancer cell lines and blocked tumor growth in a murine model of prostate cancer bone metastasis. Cell cycle studies confirmed an increase in the G1-phase and decrease in the S-phase population of cancer cells under starvation stress conditions, which correlated with elevated intracellular superoxide levels. Microarray data showed significantly decreased levels of regenerating islet-derived family member 4 (REG4 expression in prostate cancer cells with knockdown of ADAM9 gene expression. This REG4 downregulation also resulted in induction of expression of p21(Cip1/WAF1, which negatively regulates cyclin D1 and blocks the G1/S transition. Our data reveal a novel molecular mechanism of ADAM9 in the regulation of prostate cancer cell proliferation, and suggests a combined modality of ADAM9 shRNA gene therapy and cytotoxic agents for hormone refractory and bone metastatic prostate cancer.

  6. AAV-mediated expression of BAG1 and ROCK2-shRNA promote neuronal survival and axonal sprouting in a rat model of rubrospinal tract injury.

    Science.gov (United States)

    Challagundla, Malleswari; Koch, Jan Christoph; Ribas, Vinicius Toledo; Michel, Uwe; Kügler, Sebastian; Ostendorf, Thomas; Bradke, Frank; Müller, Hans Werner; Bähr, Mathias; Lingor, Paul

    2015-07-01

    A lesion to the rat rubrospinal tract is a model for traumatic spinal cord lesions and results in atrophy of the red nucleus neurons, axonal dieback, and locomotor deficits. In this study, we used adeno-associated virus (AAV)-mediated over-expression of BAG1 and ROCK2-shRNA in the red nucleus to trace [by co-expression of enhanced green fluorescent protein (EGFP)] and treat the rubrospinal tract after unilateral dorsal hemisection. We investigated the effects of targeted gene therapy on neuronal survival, axonal sprouting of the rubrospinal tract, and motor recovery 12 weeks after unilateral dorsal hemisection at Th8 in rats. In addition to the evaluation of BAG1 and ROCK2 as therapeutic targets in spinal cord injury, we aimed to demonstrate the feasibility and the limits of an AAV-mediated protein over-expression versus AAV.shRNA-mediated down-regulation in this traumatic CNS lesion model. Our results demonstrate that BAG1 and ROCK2-shRNA both promote neuronal survival of red nucleus neurons and enhance axonal sprouting proximal to the lesion.

  7. USP22 ShRNA慢病毒载体的构建及鉴定%Construction and identification of ShRNA lentivirus vector in gene USP22

    Institute of Scientific and Technical Information of China (English)

    余宏伟; 廖志伟; 庄雅靖; 喻芳; 周同冲

    2014-01-01

    目的:构建并鉴定USP22基因ShRNA慢病毒载体,为进一步研究USP22基因在鼻咽癌中的作用机制奠定基础。方法针对USP22基因的编码序列设计并合成2条特异性干扰序列,序列两端含有限制性内切酶位点HpaⅠ和X hoⅠ。寡核苷酸链退火生成寡核苷酸双链,5'端磷酸化后将含有酶切位点的寡核苷酸双链克隆到pLL3.7慢病毒表达载体。连接产物经转化、培养,提取其质粒,提取出来的质粒经HpaⅠ和XhoⅠ酶切电泳鉴定,鉴定正确的质粒进行测序。构建成功的慢病毒表达载体pLL-USP22-shRNA与包装载体质粒混匀共转染于293T细胞。通过荧光显微镜下观察绿色荧光蛋白(GFP)情况,对病毒滴度和感染效率进行检测。结果成功构建慢病毒表达载体pLL-USP22-shRNA。与包装载体质粒共转染293T细胞后测定慢病毒滴度为4×107 TU/ml。结论本实验应用相关技术成功构建USP22 ShRNA慢病毒载体,为进一步研究USP22基因的生物学功能奠定了基础。%Objective To construct and identify ShRNA lentivirus vector in gene USP22,and lay the foundation for fur-ther study of gene USP22 on mechanism of nasopharyngeal carcinoma. Methods Two special interference sequences targeting on coded sequence of gene USP22 were designed and synthesized.HpaⅠand XhoⅠas two sites of restriction enzyme were included at both ends of the sequence.Oligodeoxynucleotides was annealed to generate double strands of oligodeoxynucleotides.After 5’end phosphorylation,double strands of oligodeoxynucleotides containing restriction enzyme cutting site was cloned to pLentiLox3.7 (PLL3.7) lentiviral expression vector.The connection product was transformed and cultivated in order to extract plasmid.The extracted plasmid was identified by restriction enzyme digestion and electrophoresis,and sequencing was carried on only for correct plasmid after identification.The successfully-constructed lentiviral expression vector of p

  8. A Comparison of Target Gene Silencing using Synthetically Modified siRNA and shRNA That Express Recombinant Lentiviral Vectors.

    Science.gov (United States)

    Spirin, P V; Baskaran, D; Rubtsov, P M; Zenkova, M A; Vlassov, V V; Chernolovskaya, E L; Prassolov, V S

    2009-07-01

    RNA-interference is an effective natural mechanism of post-transcriptional modulation of gene expression. RNA-interference mechanism exist as in high eukaryotes both animals and plants as well in lower eukaryotes and viruses. RNA-interference is now used as a powerful tool in study of functional gene activity and many essential for fundamental biology results was obtained with this approach. Also it's widely believed that RNA-interference could be used in working out of new therapeutic medicine against malignant, infectious and hereditary diseases. One of the main problems of these developments is search of effective methods of siRNA transfer into the target cells. At present time for these purpose different sorts of transfect ions or viral transduction are used. At present article the results of comparison of inhibition of expression of oncogene AML-ET O by synthetic siRNA and by recombinant lentiviruses coding for corresponding shRNA are presented.

  9. Conditional expression of retrovirally delivered anti-MYCN shRNA as an in vitro model system to study neuronal differentiation in MYCN-amplified neuroblastoma

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    Tømte Ellen

    2011-01-01

    Full Text Available Abstract Background Neuroblastoma is a childhood cancer derived from immature cells of the sympathetic nervous system. The disease is clinically heterogeneous, ranging from neuronal differentiated benign ganglioneuromas to aggressive metastatic tumours with poor prognosis. Amplification of the MYCN oncogene is a well established poor prognostic factor found in up to 40% of high risk neuroblastomas. Using neuroblastoma cell lines to study neuronal differentiation in vitro is now well established. Several protocols, including exposure to various agents and growth factors, will differentiate neuroblastoma cell lines into neuron-like cells. These cells are characterized by a neuronal morphology with long extensively branched neurites and expression of several neurospecific markers. Results In this study we use retrovirally delivered inducible short-hairpin RNA (shRNA modules to knock down MYCN expression in MYCN-amplified (MNA neuroblastoma cell lines. By addition of the inducer doxycycline, we show that the Kelly and SK-N-BE(2 neuroblastoma cell lines efficiently differentiate into neuron-like cells with an extensive network of neurites. These cells are further characterized by increased expression of the neuronal differentiation markers NFL and GAP43. In addition, we show that induced expression of retrovirally delivered anti-MYCN shRNA inhibits cell proliferation by increasing the fraction of MNA neuroblastoma cells in the G1 phase of the cell cycle and that the clonogenic growth potential of these cells was also dramatically reduced. Conclusion We have developed an efficient MYCN-knockdown in vitro model system to study neuronal differentiation in MNA neuroblastomas.

  10. In vivo transfection of manganese superoxide dismutase gene or nuclear factor κB shRNA in nodose ganglia improves aortic baroreceptor function in heart failure rats.

    Science.gov (United States)

    Zhang, Dongze; Liu, Jinxu; Tu, Huiyin; Muelleman, Robert L; Cornish, Kurtis G; Li, Yu-Long

    2014-01-01

    Arterial baroreflex sensitivity is attenuated in chronic heart failure (CHF) state, which is associated with cardiac arrhythmias and sudden cardiac death in patients with CHF. Our previous study showed that CHF-induced sodium channel dysfunction in the baroreceptor neurons was involved in the blunted baroreflex sensitivity in CHF rats. Mitochondria-derived superoxide overproduction decreased expression and activation of the sodium channels in the baroreceptor neurons from CHF rats. However, the molecular mechanisms responsible for the sodium channel dysfunction in the baroreceptor neurons from CHF rats remain unknown. We tested the involvement of nuclear factor κB (NFκB) in the sodium channel dysfunction and evaluated the effects of in vivo transfection of manganese superoxide dismutase gene and NFκB shRNA on the baroreflex function in CHF rats. CHF was developed at 6 to 8 weeks after left coronary artery ligation in adult rats. Western blot and chromatin immunoprecipitation data showed that phosphorylated NFκB p65 and ability of NFκB p65 binding to the sodium channel promoter were increased in the nodose ganglia from CHF rats. In vivo transfection of adenoviral manganese superoxide dismutase gene or lentiviral NFκB p65 shRNA into the nodose ganglia partially reversed CHF-reduced sodium channel expression and cell excitability in the baroreceptor neurons and improved CHF-blunted arterial baroreflex sensitivity. Additionally, transfection of adenoviral manganese superoxide dismutase also inhibited the augmentation of phosphorylated NFκB p65 in the nodose neurons from CHF rats. The present study suggests that superoxide-NFκB signaling contributes to CHF-induced baroreceptor dysfunction and resultant impairment of baroreflex function.

  11. HIF-1α和hTERT shRNA双基因抑制脑胶质瘤生长的实验研究%The study of inhibitory effect of HIF-1α shRNA and hTERT shRNA on growth of human glioma

    Institute of Scientific and Technical Information of China (English)

    于如同; 孙卫东; 马衍刚; 黄冠程

    2008-01-01

    Objective To explore the effect of silencing Hypoxia-induced factor-1α(HIF-1α)gene and human telomerase reverse transcriptase(hTERT)gene by short hairpin RNAs(shRNAs)on growth and apoptosis of human gliomas in nude mice.Methods To cuhure human glioma U25 1 cells in vitro and establish models of Human glioma subcutaneous xenograft in nude mice.The volumes of tumors and the inhibition rates were measured five weeks after therapy.Xenograft tumors were observed with HE staining after plasmid therapy.Protein levels of HIF-1α and hTERT were detected by Western blot.Apoptosis of tunlor cells staining with Annexin V and PI were measured by flow cytometer.Resuits The subcutaneous xenograft glioma model of nude mice were constructed steadily.The volumes of tumor in HIF-1α shRNA+hTERT shRNA group was significantly decreased compared with blank plasmid group、PBS group and HIF-1α/hTERT shRNA group.the inhibition rate was 84.2%.Rare natural tumor cells and blood vessel.massive necrosie and apoptotic tumor cells were observed.Protein levels of HIF-1α and/or hTERT were also inhibited.When treated with HIF-1α shRNA and hTERT shRNA.the two protein levels were also inhibited.The apoptosic rate of tumor cells in HIF-1α shRNA+hTERT shRNA group was higher than other groups (P<0.01).Conclusion HIF-1α shRNA or hTERT shRNA could inhibite the corresponding protein levels and inhibit the Multiplication and growth of gliomas,induce apoptosis of tumor cells.Moreover,the effects of HIF-1α shRNA with hTERT shRNA is better.%目的 研究靶向缺氧诱导因子-1α(HIF-1α)和人端粒酶逆转录酶(hTERT)的短发夹RNA(shRNA),对裸鼠皮下人脑胶质瘤中HIF-1α、hTERT基因表达的抑制作用,及其对肿瘤增殖和细胞凋亡的影响.方法 体外培养人脑胶质瘤U251细胞;建立人脑胶质瘤裸鼠皮下接种模型;将HIF-1α shRNA、hTERT shRNA质粒转染入移植瘤中,观察肿瘤生长情况;HE染色法观察肿瘤组织的病理改变;Western blot

  12. 靶向 RECQL4基因的 shRNA 慢病毒表达载体的构建及鉴定

    Institute of Scientific and Technical Information of China (English)

    陈泓磊; 吴晓滨; 王华摄; 林义佳; 彭俊生

    2015-01-01

    目的:构建介导 RECQL4 RNA 干扰的 shRNA 慢病毒表达载体。方法将人工合成的 RECQL4 siR-NA 经 PCR 扩增后与线化的 pLKO.1慢病毒表达载体连接,经测序鉴定后,将 pLKO.1-RECQL4-shRNA 与慢病毒包装质粒共转染入 HEK -293T 细胞,产生慢病毒颗粒并测定病毒滴度。体外感染人结肠癌细胞 RKO 后,实时定量 PCR 和 Western blot 检测 RECQL4 mRNA 和蛋白的沉默效果。设置未加入任何慢病毒表达载体的 RKO 细胞为未感染组,感染 pLKO.1-scramble -shRNA 慢病毒载体为阴性对照组。结果基因测序结果证实成功构建 pL-KO.1-RECQL4-shRNA 慢病毒表达载体。测定包装后的慢病毒颗粒滴度为6.70×107 IU/mL。感染 RKO 细胞后,RECQL4的 mRNA 和蛋白表达量均低于阴性组和未感染组。结论成功构建靶向 RECQL4基因的 shRNA 慢病毒表达载体,可有效抑制人结肠癌 RKO 细胞 RECQL4 mRNA 和蛋白的表达。

  13. 人AQP1-shRN A 表达质粒载体的构建与筛选%Construction and screening of human AQP1 shRNA expression vectors

    Institute of Scientific and Technical Information of China (English)

    李卓; 康炜; 辛娜; 田宇; 李建华

    2015-01-01

    Objective To construct and screen effective shRNA expression vectors targeting human AQP1 gene ,and evaluate the interference efficiency of the AQP1 shRNA recombinant plasmids ,thus provide basis for further exploration on the effect and mechanism of AQP1 gene on human breast cancer cells .Methods Four pairs of shRNA sequences targeting human AQP1 gene were designed and synthesized ,and then inserted into the GV115 vector .AQP1 shRNA and control shRNA plasmids were trans‐fected into human breast cancer MCF‐7 cells .The expression of AQP1 mRNA and protein were detected by real time PCR(RT‐PCR) and Western blot to evaluate the interfering efficiency .Results RT‐PCR demonstrated that AQP1 was expressed in human breast cancer MCF‐7 cells .Sequencing showed that the shRNA vectors targeting AQP1 were successfully constructed .48 h after the AQP1 shRNA transfection ,AQP1 mRNA and protein expression levels in MCF‐7 cells were reduced to a significant degree ,and the AQP1 shRNA 4 plasmid vector could inhibit the AQP1 most efficiently .Conclusion The AQP1 shRNA recombinant plasmids vectors were successfully constructed and can significantly inhibit the expression of AQP1 in MCF‐7 human breast cancer cells .%目的:构建针对人水通道蛋白1(AQP1)基因的shRNA表达质粒载体,并验证其干扰效果,为进一步探讨AQP1基因对人乳腺癌细胞的作用及机制建立基础。方法设计合成4对针对人AQP1基因不同位点的shRNA片段,通过DNA重组技术将其插入载体GV115中,构建AQP1‐shRNA重组质粒,转染人乳腺癌MCF‐7细胞,并通过实时荧光定量 PCR(RT‐PCR)和Western blot法检测干扰效率。结果 RT‐PCR法证实AQP1在乳腺癌 MCF‐7细胞中有表达。测序验证表明4对靶向AQP1‐shRNA表达载体均构建成功。4组干扰载体能够从基因和蛋白表达水平不同程度抑制 AQP1表达,其中以AQP1‐shRNA 4对AQP1的干扰效率最强。结论成功

  14. An ShRNA Based Genetic Screen Identified Sesn2 as a Potential Tumor Suppressor in Lung Cancer via Suppression of Akt-mTOR-p70S6K Signaling.

    Directory of Open Access Journals (Sweden)

    Hui Xu

    Full Text Available Lung cancer is emerging rapidly as the leading death cause in Chinese cancer patients. The causal factors for Chinese lung cancer development remain largely unclear. Here we employed an shRNA library-based loss-of-function screen in a genome-wide and unbiased manner to interrogate potential tumor suppressor candidates in the immortalized human lung epithelial cell line BEAS-2B.Soft agar assays were conducted for screening BEAS-2B cells infected with the retroviral shRNA library with the acquired feature of anchorage-independent growth, large (>0.5mm in diameter and well-separated colonies were isolated for proliferation. PCRs were performed to amplify the integrated shRNA fragment from individual genomic DNA extracted from each colony, and each PCR product is submitted for DNA sequencing to reveal the integrated shRNA and its target gene. A total of 6 candidate transformation suppressors including INPP4B, Sesn2, TIAR, ACRC, Nup210, LMTK3 were identified. We validated Sesn2 as the candidate of lung cancer tumor suppressor. Knockdown of Sesn2 by an shRNA targeting 3' UTR of Sesn2 transcript potently stimulated the proliferation and malignant transformation of lung bronchial epithelial cell BEAS-2B via activation of Akt-mTOR-p70S6K signaling, whereas ectopic expression of Sens2 re-suppressed the malignant transformation elicited by the Sesn2 shRNA. Moreover, knockdown of Sesn2 in BEAS-2B cells promoted the BEAS-2B cell-transplanted xenograft tumor growth in nude mice. Lastly, DNA sequencing indicated mutations of Sesn2 gene are rare, the protein levels of Sesn2 of 77 Chinese lung cancer patients varies greatly compared to their adjacent normal tissues, and the low expression level of Sesn2 associates with the poor survival in these examined patients by Kaplan Meier analysis.Our shRNA-based screen has demonstrated Sesn2 is a potential tumor suppressor in lung epithelial cells. The expression level of Sesn2 may serve as a prognostic marker for

  15. Preintegration HIV-1 inhibition by a combination lentiviral vector containing a chimeric TRIM5 alpha protein, a CCR5 shRNA, and a TAR decoy.

    Science.gov (United States)

    Anderson, Joseph S; Javien, John; Nolta, Jan A; Bauer, Gerhard

    2009-12-01

    Human immunodeficiency virus (HIV) gene therapy offers a promising alternative approach to current antiretroviral treatments to inhibit HIV-1 infection. Various stages of the HIV life cycle including pre-entry, preintegration, and postintegration can be targeted by gene therapy to block viral infection and replication. By combining multiple highly potent anti-HIV transgenes in a single gene therapy vector, HIV-1 resistance can be achieved in transduced cells while prohibiting the generation of escape mutants. Here, we describe a combination lentiviral vector that encodes three highly effective anti-HIV genes functioning at separate stages of the viral life cycle including a CCR5 short hairpin RNA (shRNA) (pre-entry), a human/rhesus macaque chimeric TRIM5 alpha (postentry/preintegration), and a transactivation response element (TAR) decoy (postintegration). The major focus on designing this anti-HIV vector was to block productive infection of HIV-1 and to inhibit any formation of provirus that would maintain the viral reservoir. Upon viral challenge, potent preintegration inhibition of HIV-1 infection was achieved in combination vector-transduced cells in both cultured and primary CD34(+) hematopoietic progenitor cell (HPC)-derived macrophages. The generation of escape mutants was also blocked as evaluated by long-term culture of challenged cells. The ability of this combination anti-HIV lentiviral vector to prevent HIV-1 infection, in vitro, warrants further evaluation of its in vivo efficacy.

  16. Novel HIV-1 knockdown targets identified by an enriched kinases/phosphatases shRNA library using a long-term iterative screen in Jurkat T-cells.

    Directory of Open Access Journals (Sweden)

    Sylvie Rato

    Full Text Available HIV-1 is a complex retrovirus that uses host machinery to promote its replication. Understanding cellular proteins involved in the multistep process of HIV-1 infection may result in the discovery of more adapted and effective therapeutic targets. Kinases and phosphatases are a druggable class of proteins critically involved in regulation of signal pathways of eukaryotic cells. Here, we focused on the discovery of kinases and phosphatases that are essential for HIV-1 replication but dispensable for cell viability. We performed an iterative screen in Jurkat T-cells with a short-hairpin-RNA (shRNA library highly enriched for human kinases and phosphatases. We identified 14 new proteins essential for HIV-1 replication that do not affect cell viability. These proteins are described to be involved in MAPK, JNK and ERK pathways, vesicular traffic and DNA repair. Moreover, we show that the proteins under study are important in an early step of HIV-1 infection before viral integration, whereas some of them affect viral transcription/translation. This study brings new insights for the complex interplay of HIV-1/host cell and opens new possibilities for antiviral strategies.

  17. A Pooled shRNA Screen Identifies Rbm15, Spen, and Wtap as Factors Required for Xist RNA-Mediated Silencing

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    Benoit Moindrot

    2015-07-01

    Full Text Available X-chromosome inactivation is the process that evolved in mammals to equalize levels of X-linked gene expression in XX females relative to XY males. Silencing of a single X chromosome in female cells is mediated by the non-coding RNA Xist. Although progress has been made toward identifying factors that function in the maintenance of X inactivation, the primary silencing factors are largely undefined. We developed an shRNA screening strategy to produce a ranked list of candidate primary silencing factors. Validation experiments performed on several of the top hits identified the SPOC domain RNA binding proteins Rbm15 and Spen and Wtap, a component of the m6A RNA methyltransferase complex, as playing an important role in the establishment of Xist-mediated silencing. Localization analysis using super-resolution 3D-SIM microscopy demonstrates that these factors co-localize with Xist RNA within the nuclear matrix subcompartment, consistent with a direct interaction.

  18. Anti-cancer effect of oncolytic adenovirus-armed shRNA targeting MYCN gene on doxorubicin-resistant neuroblastoma cells.

    Science.gov (United States)

    Li, Yuan; Zhuo, Baobiao; Yin, Yiyu; Han, Tao; Li, Shixian; Li, Zhengwei; Wang, Jian

    2017-09-09

    Chemotherapy is one of the few effective choices for patients with neuroblastoma. However, the development of muti-drug resistance (MDR) to chemotherapy is a major obstacle to the effective treatment of advanced or recurrent neuroblastoma. The muti-drug resistance-associated protein (MRP), which encodes a transmembrane glycoprotein, is a key regulator of MDR. The expression of MRP is a close correlation with MYCN oncogene in neuroblastoma. We have recently shown ZD55-shMYCN (oncolytic virus armed with shRNA against MYCN) can down-regulate MYCN to inhibit tumor cells proliferation and induce apoptosis in neuroblastoma. Here we further report ZD55-shMYCN re-sensitized doxorubicin-resistant cells to doxorubicin (as shown by reduced proliferation, increased apoptosis, and inhibited cell migration), and reduced the in vivo growth rate of neuroblastoma xenografts by down-regulation of MRP expression. Sequential therapy with doxorubicin did not affect the replication of ZD55-shMYCN in doxorubicin-resistant neuroblastoma cells, but decreased the expression of Bcl-2, Bcl-XL, MMP-1. Thus, this synergistic effect of ZD55-shMYCN in combination with doxorubicin provides a novel therapy strategy for doxorubicin-resistant neuroblastoma, and is a promising approach for further clinical development. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Effects on Biological Behavior of Bladder Carcinoma T24 Cells via Silencing DNMT1 and/or DNMT3b with shRNA In Vitro

    Institute of Scientific and Technical Information of China (English)

    Shilong ZHANG; Fuqing ZENG; Shibo PENG; Chaohui ZHU; Heng LI; Liang WANG

    2009-01-01

    In this study,RNA interference technique was employed to silence the expression of DNMT1 and/or DNMT3b in human bladder cancer T24 cells.The expression levels of their mRNA and protein were greatly decreased by up to 75% and 65% respectively after T24 cells were transfected with lipofectamine2000 for 72 h,indicating RNA interference is an effective tool in gene knockdown.Proliferation and apoptosis of T24 cells were detected by MTT,and annexin-V-FITC and propidium iodide staining flow cytometry,respectively.It was found that loss of the DNMT1 or DNMT3b expression could inhibit the cell growth and promote the cell apoptosis to some extent.However,combined treatment with shRNA targeting both DNMT1 and DNMT3b mRNA could ob-viously enhance the above effects.It was concluded that simultaneously silencing both genes could result in strong suppressing effect on tumor proliferation and promoting ceil apoptosis than separate use,suggesting combined use of DNMT1 and DNMT3b can achieve a synergistic effect in the CpG island methylation in human bladder tumorigenesis.

  20. Targeting RPTPσ with lentiviral shRNA promotes neurites outgrowth of cortical neurons and improves functional recovery in a rat spinal cord contusion model.

    Science.gov (United States)

    Zhou, Heng-Xing; Li, Xue-Ying; Li, Fu-Yuan; Liu, Chang; Liang, Zhi-Pin; Liu, Shen; Zhang, Bin; Wang, Tian-Yi; Chu, Tian-Ci; Lu, Lu; Ning, Guang-Zhi; Kong, Xiao-Hong; Feng, Shi-Qing

    2014-10-24

    After spinal cord injury (SCI), the rapidly upregulated chondroitin sulfate proteoglycans (CSPGs), the prominent chemical constituents and main repulsive factors of the glial scar, play an important role in the extremely limited ability to regenerate in adult mammals. Although many methods to overcome the inhibition have been tested, no successful method with clinical feasibility has been devised to date. It was recently discovered that receptor protein tyrosine phosphatase sigma (RPTPσ) is a functional receptor for CSPGs-mediated inhibition. In view of the potential clinical application of RNA interference (RNAi), here we investigated whether silencing RPTPσ via lentivirus-mediated RNA interference can promote axon regeneration and functional recovery after SCI. Neurites of primary rat cerebral cortical neurons with depleted RPTPσ exhibited a significant enhancement in elongation and crossing ability when they encountered CSPGs in vitro. A contusion model of spinal cord injury in Wistar rats (the New York University (NYU) impactor) was used for in vivo experiments. Local injection of lentivirus encoding RPTPσ shRNA at the lesion site promoted axon regeneration and synapse formation, but did not affect the scar formation. Meanwhile, in vivo functional recovery (motor and sensory) was also enhanced after RPTPσ depletion. Therefore, strategies directed at silencing RPTPσ by RNAi may prove to be a beneficial, efficient and valuable approach for the treatment of SCI. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Kinome-wide shRNA Screen Identifies the Receptor Tyrosine Kinase AXL as a Key Regulator for Mesenchymal Glioblastoma Stem-like Cells

    Directory of Open Access Journals (Sweden)

    Peng Cheng

    2015-05-01

    Full Text Available Glioblastoma is a highly lethal cancer for which novel therapeutics are urgently needed. Two distinct subtypes of glioblastoma stem-like cells (GSCs were recently identified: mesenchymal (MES and proneural (PN. To identify mechanisms to target the more aggressive MES GSCs, we combined transcriptomic expression analysis and kinome-wide short hairpin RNA screening of MES and PN GSCs. In comparison to PN GSCs, we found significant upregulation and phosphorylation of the receptor tyrosine kinase AXL in MES GSCs. Knockdown of AXL significantly decreased MES GSC self-renewal capacity in vitro and inhibited the growth of glioblastoma patient-derived xenografts. Moreover, inhibition of AXL with shRNA or pharmacologic inhibitors also increased cell death significantly more in MES GSCs. Clinically, AXL expression was elevated in the MES GBM subtype and significantly correlated with poor prognosis in multiple cancers. In conclusion, we identified AXL as a potential molecular target for novel approaches to treat glioblastoma and other solid cancers.

  2. An ShRNA Screen Identifies MEIS1 as a Driver of Malignant Peripheral Nerve Sheath Tumors

    Directory of Open Access Journals (Sweden)

    Ami V. Patel

    2016-07-01

    Full Text Available Malignant peripheral nerve sheath tumors (MPNST are rare soft tissue sarcomas that are a major source of mortality in neurofibromatosis type 1 (NF1 patients. To identify MPNST driver genes, we performed a lentiviral short hairpin (sh RNA screen, targeting all 130 genes up-regulated in neurofibroma and MPNSTs versus normal human nerve Schwann cells. NF1 mutant cells show activation of RAS/MAPK signaling, so a counter-screen in RAS mutant carcinoma cells was performed to exclude common RAS-pathway driven genes. We identified 7 genes specific for survival of MPSNT cells, including MEIS1. MEIS1 was frequently amplified or hypomethylated in human MPSNTs, correlating with elevated MEIS1 gene expression. In MPNST cells and in a genetically engineered mouse model, MEIS1 expression in developing nerve glial cells was necessary for MPNST growth. Mechanistically, MEIS1 drives MPNST cell growth via the transcription factor ID1, thereby suppressing expression of the cell cycle inhibitor p27Kip and maintaining cell survival.

  3. Recombinant adenovirus-mediated shRNA silencing of midkine gene in BxPC-3 cells

    Institute of Scientific and Technical Information of China (English)

    Mingyue Xiong; Kunzheng Wang

    2009-01-01

    Objective:To investigate the silencing effects of recombinant adenovirus Ad-shRNA-MK on midkine(MK) gene in pancreatic cancer cells. Methods:Ad-shRNA-MK was used to infect pancreatic cancer BxPC-3 cells. Assays were conducted for knockdown of the MK gene on the day of infection and on the 1a, 3rd, 5th, 7th, and 9th days post-infection by using immunocytochemistry, real-time RT-PCR, and Western blot analysis. Results:The adenoviral Ad-shRNA-PTN was constructed successfully, and infection was confirmed by electron microscopic observation. By using real-time RT-PCR, the inhibition rates of MK mRNA expression in the BxPC-3 cells were 20%, 80%, 55%, and 23% on the 1st, 3rd, 5th, and 7th days post-infection. Immunocytochemistry and Western blot analysis confirmed this effect at the gene product level. Conclusion:Efficient and specific knockdown of MK in pancreatic cancer cells by adenoviral Ad-shRNA-PTN is a potentially powerful tool for the study of gene therapy of pancreatic cancer nerve infiltration.

  4. Silencing of hepatitis C virus replication by a non-viral vector based on solid lipid nanoparticles containing a shRNA targeted to the internal ribosome entry site (IRES).

    Science.gov (United States)

    Torrecilla, Josune; Del Pozo-Rodríguez, Ana; Solinís, María Ángeles; Apaolaza, Paola S; Berzal-Herranz, Beatriz; Romero-López, Cristina; Berzal-Herranz, Alfredo; Rodríguez-Gascón, Alicia

    2016-10-01

    Gene silencing mediated by RNAi has gained increasing interest as an alternative for the treatment of infectious diseases such as refractory hepatitis C virus (HCV) infection. In this work we have designed and evaluated a non-viral vector based on solid lipid nanoparticles (SLN) bearing hyaluronic acid, protamine and a short hairpin RNA (shRNA74) targeted to the Internal Ribosome Entry Site (IRES) of the HCV. The vector was able to inhibit the expression of the HCV IRES in Huh-7 cells, with the inhibition level dependent on the shRNA74 to SLN ratio and on the shRNA74 dose added to the culture cells. The nanocarrier was also able to inhibit the replication in human hepatoma cells supporting a subgenomic HCV replicon (Huh-7 NS3-3'). The vector was quickly and efficiently internalized by the cells, and endocytosis was the most productive uptake mechanism for silencing. Clathrin-mediated endocytosis and to a lesser extent caveolae/lipid raft-mediated endocytosis were identified as endocytic mechanisms involved in the cell uptake. Internalization via the CD44 receptor was also involved, although this entry route seems to be less productive for silencing than endocytosis. The vector did not induce either hemolysis or agglutination of red cells in vitro, which was indicative of good biocompatibility. In summary, we have shown for the first time the ability of a non-viral SLN-based vector to silence a HCV replicon.

  5. Targeted P2X7 R shRNA delivery attenuates sympathetic nerve sprouting and ameliorates cardiac dysfunction in rats with myocardial infarction.

    Science.gov (United States)

    Gao, Hongmei; Yin, Jie; Shi, Yugen; Hu, Hesheng; Li, Xiaolu; Xue, Mei; Cheng, Wenjuan; Wang, Ye; Li, Xinran; Li, Yongkang; Wang, Yu; Yan, Suhua

    2017-04-01

    Inflammation-dominated sympathetic sprouting adjacent to the necrotic region following myocardial infarction (MI) has been implicated in the etiology of arrhythmias resulting in sudden cardiac death; however, the mechanisms responsible remain to be elucidated. Although P2X7 R is a key immune mediator, its role has yet to be explored. We investigated whether P2X7 R regulates NF-κB and affects cardiac sympathetic reinnervation in rats undergoing MI. An adenoviral vector with a short hairpin RNA (shRNA) sequence inserted was adopted for the inhibition of P2X7 R in vivo. Myocardial infarction was induced by left coronary artery ligation, and immediately after that, recombinant P2X7 R-shRNA adenovirus, negative adenovirus (control), or normal saline solution (vehicle) was injected intramyocardially around the MI region and border areas. A high level of P2X7 R was activated in the infarcted tissue at an early stage. The administration of P2X7 R RNAi resulted in the inhibition of Akt and Erk1/2 phosphorylation and decreased the activation of NF-κB and macrophage infiltration, as well as attenuated the expression of nerve growth factor (NGF). Eventually, the NGF-induced sympathetic hyperinnervation was blunted, as assessed by the immunofluorescence of tyrosine hydroxylase (TH) and growth-associated protein 43 (GAP 43). At 7 days post-MI, the arrhythmia score of programmed electrical stimulation in the vehicle-treated infarcted rats was higher than the MI-shRNA group. Further amelioration of cardiac dysfunction was also detected. The administration of P2X7 R RNAi during the acute inflammatory response phase prevented the process of sympathetic hyperinnervation after MI, which was associated in part with inhibiting the Akt and ERK1/2 pathways and NF-κB activation. © 2016 John Wiley & Sons Ltd.

  6. ROCK1 and ROCK2 silencing by ShRNA in rat cardiomyocytes%ShRNA对大鼠心肌细胞ROCK1和ROCK2表达的沉默作用

    Institute of Scientific and Technical Information of China (English)

    丁浩; 李菊香; 孙国芳; 洪葵; 程晓曙

    2011-01-01

    背景:已有研究发现,ROCK1和ROCK2参与了细胞的凋亡过程,由此假设下调ROCK1和ROCK2表达能抑制心肌细胞凋亡.目的:验证ROCK1-shRNA和ROCK2-shRNA转染能否有效沉默大鼠心肌细胞内ROCK1和ROCK2基因的表达,并筛选沉默效率最高的shRNA.方法:以ROCK1和ROCK2为靶基因,分别构建了3个特异性shRNA重组质粒,转染这些shRNA后观察其对SD大鼠心肌细胞中ROCK1和ROCK2的沉默作用,并筛选出沉默效率最佳的shRNA.结果与结论:Western Blot结果显示,与对照组相比,3个重组质粒ROCK1-shRNA和ROCK2-shRNA的转染均能明显降低SD大鼠心肌细胞中ROCK1和ROCK2的表达(P < 0.05或P < 0.01),其中ROCK1-shRNA1和ROCK2-shRNA2的沉默效率最高(P < 0.05),分别为(82.15±7.53)%和(89.42±5.81)%.这表明ROCK1-shRNA和ROCK2-shRNA转染能有效沉默大鼠心肌细胞内ROCK1和ROCK2基因的表达,并筛选出了沉默效率最高的shRNA.%BACKGROUND: Previous studies have proved that Rho-associated coiled-coilproteinkinase(ROCK) 1 and ROCK2 invoke indown-regulating the expression ofROCKI andROCK2.OBJECT FVE: To verify whether the transfection of ROCK1-short hairpin R HA (ShRHA) and ROCK2-shRHA can effective^silence the expressions ofROCKI and ROCK2 inrat car d io my ocytes and to screen for the shR HA with the best silencingeffeciency.METHODS: Three specific recombinant plasmids of ROC K1-shRH A and ROCK2-shRHA were coretructed and identifiedrespectively. Silencing effects of ROCK1 and ROCK2 in SD rat myocardial cells were assessed after shRHA trarEfection. Andthe shRHA with the best silencing eff I cie n cy w as selected among these shRHAs.RESULTS AHD COHCL USIOHS: Western blot results showed that, compared with the control groups, the transfection of allthree recombinant plasmids of ROCK1-shRHAs and ROCK2-shRHAs inhibited the expressions of ROCK1 and ROCK2significantly^ < 0.05 or P < 0.01). Among these shRHAs. ROCK1-shRHA1 and ROCK2-shRHA2 showed the best silencingefficiency of

  7. Construction and package of recombinant lentivirus expressing shRNA targeting TIPE2 gene%靶向TIPE2基因的shRNA慢病毒载体构建及病毒包装

    Institute of Scientific and Technical Information of China (English)

    王耀辉; 王明丽; 陈九格; 张晶; 马远方

    2014-01-01

    目的:包装表达TIPE2-shRNA的慢病毒颗粒。方法查询并合成7对针对TIPE2的shRNA序列,将其克隆入pLK01.载体,经酶切及测序正确后,将构建好的7种pLKO 1.-TIPE2-shRNA质粒分别与 pCDNA31./3× FLAG-TIPE2共转染293T细胞,Western blot鉴定干扰效率并进行后续的病毒包装。结果4号质粒沉默效果最好,将其与 Non-target-shRNA质粒分别进行病毒包装,Western blot和免疫荧光证实病毒具有良好的感染效率和沉默效果。结论 TIPE2-shRNA的慢病毒载体构建及病毒包装成功。%Objective To package the recombinant lentivirus expressing TIPE2 shRNA .Methods Inquiring and synthesizing seven pairs shRNA sequences targeting TIPE2 ,then inserted into pLKO .1 lentivirus vector .After the constructions were verified to be correct ,co-transfected them with pCDNA3 .1/3 × FLAG TIPE2 plasmids into 293T cells , respectively . Then , Non target shRNA , the highest silence efficiency plasmid were co-transfected with psPAX2 , pMD2 .G into 293T cells for lentivirus packaging , respectively .Results The No .4 plasmid was screened by Western blotting that has the highest silence efficiency and followed by virus packing . The infection and silence efficiency was confirmed by immunofluorescence and Western blot . Conclusion The recombinant lentivirus expressing shRNA targeting TIPE2 gene was packaged successfully .

  8. 人神经菌毛素-1基因shRNA慢病毒载体的构建%Construction of recombinant lentiviral vector containing shRNA for human neuropilin-1 gene

    Institute of Scientific and Technical Information of China (English)

    郑见宝; 耿智敏; 陈强; 王林

    2012-01-01

    AIM; To construct and identify a recombi-nant lentiviral vector containing shRNA for human neuropilin-1 (NRP-1) gene. METHODS: Four shRNA targeting the NRP-1 mRNA were designed to construct the pGCSIL-RFP-shNRP1 lentivirus vectors. The positive clone was chosen and confirmed by PCR and DNA sequencing. 293T cells were cotransfected with pGCSIL-RFP-shNRP1, pHelperl.0 and pHelper 2.0 to package the lentivirus and the titer of the virus was tested. After lentivirus-shRNA and over-expression plasmid containing NRP-1 were transfected into 293T cells, Western blotting was used to determine the expression of Flag gene in order to observe the inhibited efficacy of relative NRP-1 expression. RESULTS: PCR analysis and DNA sequencing demonstrated that the shRNA sequence was consistent with the human NRP-1. The titer of the recombinant lentiviral vector was 1×10-9 Tu/mL The relative expression of NRP-1 protein in the transfected cells significantly decreased after treated with lentiviral-shRNA. CONCLUSION: We have constructed successfully the effective recombinant lentiviral vector containing shRNA for human NRP-1 gene.%目的:构建并鉴定靶向人神经菌毛素-1(NRP-1)基因的小发卡RNA的慢病毒载体.方法:针对NRP-1mRNA设计了4条shRNA,并构建pGCSIL-RFP-shNRP1慢病毒质粒,PCR扩增阳性克隆并测序鉴定.用pGCSIL-RFP-shNRP1、pHelper1.0和pHelper2.0质粒共转染293T细胞包装产生慢病毒,测定病毒滴度.将慢病毒干扰RNA及含有NRP-1过表达载体共转染293T细胞,Western blot法检测Flag表达,观察NRP-1蛋白相对表达抑制效果.结果:PCR和测序结果与设计的干扰序列一致,病毒滴度达1×109 Tu/mL.转染细胞中NRP-1蛋白相对表达显著降低.结论:成功构建了高效率的人NRP-1基因shRNA慢病毒载体.

  9. Designing Ago2-specific siRNA/shRNA to Avoid Competition with Endogenous miRNAs.

    Science.gov (United States)

    Ma, Hongming; Zhang, Junli; Wu, Haoquan

    2014-07-15

    Relatively large amounts of transfected siRNA can compete for Ago proteins and thus compromise endogenous miRNA function, potentially leading to toxicities. Here, we show that shRNA can also perturb endogenous miRNA function similarly. More importantly, we also show that the problem can be solved by designing shRNAs in the context of pre-miR-451 structure with completely complementary stem, which significantly improves the Ago2 specificity. This shRNA was shown to be Ago2-specific, and maintain target-silencing ability while avoiding competition with endogenous miRNAs by not competing for Agos 1, 3, and 4. We conclude that modified pre-miR-451 structure provides a general platform to design shRNAs that significantly reduce perturbation of miRNA function.

  10. 大鼠野生型TRAF6基因和TRAF6shRNA表达质粒的构建及鉴定%Construction and identification of TRAF6 gene and its shRNA expression vector

    Institute of Scientific and Technical Information of China (English)

    邱文; 单锴; 庞蓉蓉; 季明德; 王迎伟

    2011-01-01

    Objective: To construct tumor necrosis factor receptor-associated factor 6 (TRAF6) and its small hair RNA (shRNA) expression vectors, and to assess their function on rat glomerular mesangial cell (CMC). Methods: The full-length TRAF6 CDS (HA tag) or three kinds of shRNAs targeting TRAF6 gene were synthesized and cloned into eukaryotic expression plasmid pcDNA3.1 and pGenesil-1/GFP,respectively. After confirmed by restriction endonuclease digestion and nucleotide sequencing,the recombinant plasmids were transfected into cultured rat GMC through Neon? Transfection system. Western blot assay was then used to confirm the expression of HA-TRAF6 and find out the optimal shRNA against TRAF6 gene. Results; It was verified by restriction endonuclease digestion and nucleotide sequencing that the constructed eukaryotic vectors were all correct. Western blot assay showed that the plasmids of pcDNA3.1-HA-TRAF6 could express HA-TRAF6 and the TRAF6 shRNA-1 (shTRAF6-l) was able to silence the target gene effectively. Conclusion; The expression plasmid of TRAF6 gene and its shRNA were constructed successfully, which provides essential experimental tools for studying biological functions of TRAF6 gene in the future.%目的:构建大鼠野生型肿瘤坏死因子受体相关因子6 (tumor necrosis factor receptor-associated factor 6,TRAF6)基因及其特异性短发卡状小干涉RNA (shRNA)真核表达质粒,并观察其在大鼠肾小球系膜细胞(GMC)中过表达和沉默TRAF6基因的情况.方法:用DNA重组技术将针对大鼠TRAF6基因的CDS区序列(加HA标签)和针对其不同位点所设计的3种shRNA序列分别克隆到pcDNA3.1及pGenesil- 1/GFP真核表达质粒中.在酶切鉴定及序列测定正确后,用NeonTM电转仪,将上述质粒分别转染入培养的大鼠GMC中,然后用Western blot检查HA-TRAF6融合蛋白的表达情况,同时筛选最有效的TRAF6shRNA.结果:限制性酶切及核酸序列分析确证,上述TRAF6基因过表达和shRNA

  11. Interference of Caspase-3 shRNA on neural cells apoptosis induced by aluminum%Caspase-3 shRNA对染铝神经细胞凋亡的干预

    Institute of Scientific and Technical Information of China (English)

    张勤丽; 教霞; 徐丽; 李娜; 牛侨

    2012-01-01

    目的 通过体外实验来研究铝的神经毒性,探讨慢病毒载体Caspase-3 RNA于扰对染铝神经细胞凋亡的干预效果.方法 原代培养神经细胞,用AlCl3·6H2O染毒,选用病毒为载体的Caspase-3 shRNA试剂感染神经细胞,荧光标记感染成功的细胞计算神经细胞的感染效率,荧光定量PCR技术检测干预后Caspase-3基因的表达情况计算慢病毒载体的Caspase-3 shRNA的抑制率.光学显微镜、荧光染色观察神经细胞的形态学变化,并检测细胞活力;AnnexinV-PI双染法检测神经细胞感染后的凋亡坏死率.结果 慢病毒载体的Caspase-3 RNA干扰试剂的感染效率大于90%,抑制率为66.47%.用慢病毒载体的Caspase-3 shRNA感染原代培养神经细胞后可见染铝神经细胞活力显著上升(P<0.01);吖啶橙(AO)-溴化乙啶(EB)双荧光染色法可以清楚地观察到染铝细胞的早期凋亡和晚期凋亡现象明显减少;Annexin V-PI双染法检测结果显示,感染后染铝神经细胞凋亡率显著降低(P<0.01).结论 馒病毒载体的Caspase-3shRNA感染细胞后,可以显著抑制Caspase-3基因的表达,神经细胞活力明显增高,凋亡细胞的数量显著降低.%Objective To study the neurotoxicity of aluminum ( Al) , and explore the intervention effect of Caspase-3 shRNA on Al-induced apoptosis. Methods Primary cultured nerve cells were exposed to AlCl3 -6H2O, Caspase-3 shRNAs were infected in the cells. Fluorescent labeled shRNAs were used to label the successfully infected cells, and infectious rates were calculated. Interference rate of Caspase-3 shRNAs on Caspase-3 gene expression was calculated with QRT-PCR method. Morphologic characteristics of Caspase-3 shRNAs treated neural cells were observed under light microscope, and fluorescent microscope stained with AO-EB staining, the alternations of apoptosis rates were analyzed by cytometry. Results The infectious rate of Caspase-3 shRNA was above 90% , the inhibition rate was 66

  12. Severe Nephrotoxic Nephritis following Conditional and Kidney-Specific Knockdown of Stanniocalcin-1.

    Directory of Open Access Journals (Sweden)

    Luping Huang

    Full Text Available Inflammation is the hallmark of nephrotoxic nephritis. Stanniocalcin-1 (STC1, a pro-survival factor, inhibits macrophages, stabilizes endothelial barrier function, and diminishes trans-endothelial migration of leukocytes; consistently, transgenic (Tg overexpression of STC1 protects from nephrotoxic nephritis. Herein, we sought to determine the phenotype of nephrotoxic nephritis after conditional and kidney-specific knockdown of STC1.We used Tg mice that, express either STC1 shRNA (70% knockdown of STC1 within 4d or scrambled shRNA (control upon delivery of Cre-expressing plasmid to the kidney using ultrasound microbubble technique. Sheep anti-mouse GBM antibody was administered 4d after shRNA activation; and mice were euthanized 10 days later for analysis.Serum creatinine, proteinuria, albuminuria and urine output were similar 10 days after anti-GBM delivery in both groups; however, anti-GBM antibody delivery to mice with kidney-specific knockdown of STC1 produced severe nephrotoxic nephritis, characterized by severe tubular necrosis, glomerular hyalinosis/necrosis and massive cast formation, while control mice manifested mild tubular injury and crescentic glomerulonephritis. Surprisingly, the expression of cytokines/chemokines and infiltration with T-cells and macrophages were also diminished in STC1 knockdown kidneys. Staining for sheep anti-mouse GBM antibody, deposition of mouse C3 and IgG in the kidney, and antibody response to sheep IgG were equal.nephrotoxic nephritis after kidney-specific knockdown of STC1 is characterized by severe tubular and glomerular necrosis, possibly due to loss of STC1-mediated pro-survival factors, and we attribute the paucity of inflammation to diminished release of cytokines/chemokines/growth factors from the necrotic epithelium.

  13. Construction of eukaryotic vector expressing shRNA of ovarian cancer associated antigen CD147%CD147 shRNA真核表达质粒的构建及鉴定

    Institute of Scientific and Technical Information of China (English)

    邹伟; 辛晓燕; 杨红; 侯向华

    2005-01-01

    目的针对CD147基因的不同部位,构建不同CD147 shRNA表达质粒载体,在转录后水平抑制CD147的表达,对其克隆进行鉴定并挑选出抑制效率最高的克隆.方法用DNA重组技术将针对人CD147基因的不同部位所设计的3对shRNA序列克隆到真核表达质粒pGE-1中,构建CD147 shRNA表达质粒载体pGE-1 CD147shRNA1、2、3.用阳离子脂质体介导转染人卵巢癌高转移细胞系HO-8910pm,经G418筛选后获得克隆进行鉴定.结果3个CD147 shRNA表达载体pGE-1CD147shRNA1、2、3经PCR、限制性酶切及部分序列分析证明基因插入正确.半定量RT-PCR、Western blotting均证实:转染细胞8910 pm/pGE-1 CD147shRNA,与亲本细胞相比,CD147的表达在mRNA和蛋白水平均明显下降.结论成功构建了CD147 shRNA表达载体pGE-1 CD147shRNA,并筛选出特异而高效地阻断CD147表达的克隆.此实验结果为进一步研究CD147蛋白分子的生物学功能及应用奠定了基础,并对shRNA设计靶序列的选择有一定指导意义.

  14. 靶向 Pokemon 基因 shRNA 重组质粒的构建及其在 CNE2细胞的筛选%Construction of short hairpin RNA recombinant plasmids targeting human Pokemon gene and screening in CNE2 cells

    Institute of Scientific and Technical Information of China (English)

    刘斐; 焦伟; 蓝娇; 肖瑞平

    2014-01-01

    Objective:To construct three short hairPin RNA( shRNA)recombinant Plasmids targeting human Pokemon gene and screen a Plasmid which can block the Pokemon exPression at maximal degree in nasoPharyngeal carcinoma( NPC)CNE2 cells. Methods:Three shRNAs were designed according to the coding sequences of the Pokemon gene. The shRNA sequences were inserted into PRNAT - U6. 1 / Neo during restriction enzyme,connection, transformation and DNA sequencing. The recombinant Plasmids and blank Plasmid were transfected into CNE2 cells using LiPofectamine 2000. The exPression of Pokemon mRNA and Protein was detected by real time Polymerase chain reaction(PCR)and Western blot,the transfected blank Plasmid CNE2 was used as a negative control,the untreated CNE2 was used as a blank control. Results:The results of DNA sequencing showed shRNA fragments were accordance with the designed sequences. ComParing with the negative control,the Pokemon mRNA and Protein exPression levels of the shRNA grouPs were down - regulated. The Pokemon results of shRNA2 grouP was lowest. Conclusion:Construc-tion and screening shRNA recombinant Plasmids targeting human Pokemon gene can Provide an effective tool for fur-ther studying the role of Pokemon gene in occurrence and develoPment of NPC.%目的:构建3个靶向 Pokemon 基因的短发夹 RNA(shRNA)真核表达载体,在鼻咽癌细胞株 CNE2中筛选出干扰效果最佳的载体。方法:依据 shRNA 设计原则,针对 Pokemon mRNA 序列设计并化学合成3对RNA 干扰序列,经酶切、连接和转化,插入 PRNAT - U6.1/ Neo 质粒,经 DNA 测序验证。脂质体介导 shRNA 重组质粒及空白质粒转染 CNE2细胞,荧光定量 PCR 和 Western blot 检测 Pokemon 在 mRNA 和蛋白水平的表达,转染空白质粒的 CNE2作为阴性对照,未加处理的 CNE2作为空白对照。结果:DNA 测序结果显示 shRNA片段与设计序列一致;转染各重组质粒后的 CNE2细胞

  15. The interference effects on UL29 shRNA expression vectors on HSV-2%单纯疱疹Ⅱ型病毒UL29基因shRNA表达载体的干扰效应

    Institute of Scientific and Technical Information of China (English)

    黄畅; 潘晓瑜; 袁俊杰; 吕延成

    2014-01-01

    目的:应用RNA干扰技术,针对单纯疱疹Ⅱ型病毒(HSV-2)UL29基因构建短发夹RNA重组表达载体,观察其对HSV-2的干扰效应。方法:针对HSV-2 UL29基因筛选出4条拟干扰靶位点序列,分别设计、合成4组靶向UL29基因shRNA的基因真核表达载体。通过脂质体转染到HEK293细胞。再将HSV-2接种到HEK293细胞中。采用终点滴定法检测病毒滴度,RT-PCR检测shRNA对转录水平的影响,Western-blot检测shRNA对蛋白质表达水平的影响。结果:终点滴定法结果显示UL29shRNA各组均可不同程度降低病毒感染滴度,与空白组(未转染重组表达载体)比较差异显著(P<0.01)。RT-PCR结果显示,与空白组比较,4组抑制率分别为28.80%、59.95%、66.08%、36.27%,差异均具有显著性(P<0.05),shRNANC(不针对任何基因的序列)与空白组相比差异无显著性,其中以UL29shRNA1461组效果为最佳。 Western-blot检测表明UL29shRNA各组ICP8蛋白表达水平比阴性对照组明显降低。结论:UL29shRNA能有效干扰HSV-2UL29基因的表达,抑制HSV-2在HEK293细胞中的复制。%Objective To construct short hairpin RNA (shRNA) recombinant expression vector for herpes simplex virus typeⅡ(HSV-2) UL29 gene and observe its inhibitory effect on HSV-2. Methods Four interference target sites of HSV-2UL29 gene were selected to construct 4 groups of small hairpin RNA respectively,named shRNA recombinant expression vector. The expression vectors were transfected into HEK293 cells with liposome. HEK293 cells were infected with HSV-2 after expression vector being transfected. The viral titer was estimated by end-point titration assay. The level of transcription was estimated by Real-Time PCR method. The expressing effect of protein was detected by Western-blot. Results Recombinant expression vector pGPU6/GFP/Neo-shRNA was constructed successfully. The result of end-point titration assay

  16. Potent and specific inhibition of SARS-CoV antigen expression by RNA interference

    Institute of Scientific and Technical Information of China (English)

    TAO Peng; ZHANG Jun; TANG Ni; ZHANG Bing-qiang; HE Tong-chuan; HUANG Ai-long

    2005-01-01

    Background Severe acute respiratory syndrome (SARS) is an infectious disease caused by SARS-CoV. There are no effective antiviral drugs for SARS although the epidemic of SARS was controlled. The aim of this study was to develop an RNAi (RNA interference) approach that specifically targeted the N gene sequence of severe acute respiratory syndrome associated coronavirus (SARS-CoV) by synthesizing short hairpin RNA (shRNA) in vivo, and to assess the inhibitory effect of this shRNA on SARS-CoV N antigen expression. Methods The eukaryotic expression plasmid pEGFP-C1-N, containing SARS-CoV N gene, was co-transfected into 293 cells with either the RNAi plasmid pshRNA-N or unrelated control plasmid pshRNA-HBV-C4. At 24, 48 and 72 hours post transfection, the green fluorescence was observed through a fluorescence microscope. The RNA levels of SARS-CoV N were determined by reverse transcription polymerase chain reaction (RT-PCR). The expression of Green Fluorescent Protein (GFP) and protein N were detected using Western blot.Results The vector, pshRNA-N expressing shRNA which targeted the N gene of SARS-CoV, was successfully constructed. The introduction of RNAi plasmid efficiently and specifically inhibited the synthesis of protein N. RT-PCR showed that RNAs of N gene were clearly reduced when the pEGFP-C1-N was cotransfected with pshRNA-N, whereas the control vector did not exhibit inhibitory effect on N gene transcription.Conclusions Our results demonstrate that RNAi mediated silencing of SARS-CoV gene could effectively inhibit expression of SARS-CoV antigen, hence RNAi based strategy should be further explored as a more efficacious antiviral therapy of SARS-CoV infection.

  17. Sequence-non-specific effects generated by various types of RNA interference triggers.

    Science.gov (United States)

    Olejniczak, Marta; Urbanek, Martyna O; Jaworska, Edyta; Witucki, Lukasz; Szczesniak, Michal W; Makalowska, Izabela; Krzyzosiak, Wlodzimierz J

    2016-02-01

    RNA interference triggers such as short interfering RNA (siRNA) or genetically encoded short hairpin RNA (shRNA) and artificial miRNA (sh-miR) are widely used to silence the expression of specific genes. In addition to silencing selected targets, RNAi reagents may induce various side effects, including immune responses. To determine the molecular markers of immune response activation when using RNAi reagents, we analyzed the results of experiments gathered in the RNAimmuno (v 2.0) and GEO Profiles databases. To better characterize and compare cellular responses to various RNAi reagents in one experimental system, we designed a reagent series in corresponding siRNA, D-siRNA, shRNA and sh-miR forms. To exclude sequence-specific effects the reagents targeted 3 different transcripts (Luc, ATXN3 and HTT). We demonstrate that RNAi reagents induce a broad variety of sequence-non-specific effects, including the deregulation of cellular miRNA levels. Typical siRNAs are weak stimulators of interferon response but may saturate the miRNA biogenesis pathway, leading to the downregulation of highly expressed miRNAs, whereas plasmid-based reagents induce known markers of immune response and may alter miRNA levels and their isomiR composition.

  18. Specific Phobias

    Science.gov (United States)

    ... Mental Health This information in Spanish ( en español ) Specific phobias Treatment More information on specific phobias A specific ... targeted psychotherapy. Return to top More information on Specific phobias Explore other publications and websites Phobias (Copyright © American ...

  19. Neuron-specific RNA interference using lentiviral vectors

    DEFF Research Database (Denmark)

    Nielsen, Troels Tolstrup; Marion, Ingrid van; Hasholt, Lis

    2009-01-01

    BACKGROUND: Viral vectors have been used in several different settings for the delivery of small hairpin (sh) RNAs. However, most vectors have utilized ubiquitously-expressing polymerase (pol) III promoters to drive expression of the hairpin as a result of the strict requirement for precise...... transcriptional initiation and termination. Recently, pol II promoters have been used to construct vectors for RNA interference (RNAi). By embedding the shRNA into a micro RNA-context (miRNA) the endogenous miRNA processing machinery is exploited to achieve the mature synthetic miRNA (smiRNA), thereby expanding...... the possible promoter choices and eventually allowing cell type specific down-regulation of target genes. METHODS: In the present study, we constructed lentiviral vectors expressing smiRNAs under the control of pol II promoters to knockdown gene expression in cell culture and in the brain. RESULTS: We...

  20. Inhibition of cervical cancer cell growth in vitro and in vivo by lentiviral-vector mediated shRNA targeting the common promoter of HPV16 E6 and E7 oncogenes.

    Science.gov (United States)

    Zhou, Jiansong; Li, Baohua; Peng, Chanjuan; Wang, Fenfen; Fu, Zhiqin; Zhou, Caiyun; Hong, Die; Ye, Feng; Lü, Weiguo; Xie, Xing

    2013-05-01

    Deregulated expression of high-risk human papillomavirus oncogenes (E6 and E7) is a pivotal event for pathogenesis and progression in cervical cancer. Both viral oncogenes are therefore regarded as ideal therapeutic targets. Small interfering RNAs (siRNA) or double-stranded RNAs can knock down target genes effectively through siRNA-induced transcriptional gene silencing (TGS). Here, we established lentiviral-vector mediated shRNA (LV-shRNA) targeting common promoter of HPV16 E6/E7 and targeting E6 transcript, transduced the lentiviral construct into cervical HPV16-positive cell lines Siha and Caski, then selected and established stably transduced monoclonal cell lines. The results showed that LV-shRNA targeting promoter, as well as targeting E6 transcript, effectively knocked down E6 and E7 expression, resulted in accumulation of p53 and pRB protein and decrease of MCM7 and p16 protein, and consequently remarkably reduced the abilities of proliferation and invasiveness of cervical cancers cells in vitro. Then we inoculated subcutaneously those monoclonal cells into nude mice to establish the transplanted tumor animal models, and found dramatically inhibited tumorigenesis and growth, as well as prolonged survival time of mice incubated by cells with LV-shRNA targeting promoter and E6 transcript. Our results may provide evidence for application of LV-shRNA targeting HR-HPV key oncogenes, as a new treatment strategy, in cervical and other HPV-associated cancer therapy. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. shRNA抑制人RPE细胞中HIF-1α表达下调后IGF-1对VEGF表达的影响%Effect of IGF-1 on expressions of VEGF under suppression of HIF-1α by shRNA in cultured human RPE cells

    Institute of Scientific and Technical Information of China (English)

    吕明良; 李敏

    2012-01-01

    Background Proliferative vitreo-retinal disease (PVD)is one group of ocular complications marked by the enhanced proliferation of various cells included retinal pigment epithelial (RPE) cells.Insulin-like growth factor-1 (IGF-1)and vascular endothelial growth factor(VEGF) are implicated in the aberrant cell proliferation and pathological neovascularization that characterizes PVD,but the signaling mechanism is unclear now. Objective This study was to explore the effect of IGF-1 on VEGF in cultured human RPE cells under the small hairpin loop RNA (shRNA) keeping hypoxia-inducible factor-1 α ( HIF-1 α) silencing. Methods Human retinas were isolated from 4 healthy male donors,and the RPE cells were harvested and cultured.The ceils were identified using anti-human keratin antibody.The third to fifth generation of human RPE cells were used in the experiment.One target site of HIF-1α mRNA was chosen by certain design principle,and shRNA was designed and synthesized by the target site and transferred into the cells in vitro,and then the cells were cultivated with 50 μg/L IGF-1 for 24 hours.The mRNA and protein expressions of HIF-1α and VEGF were detected by RT-PCR and Western blot respectively. Results Cultured human RPE cells showed the flat irregularly multangular shape,and 97% cells appeared the positive response for keratin.HIF-1α mRNA expression in human RPE cells was significantly lower in 50 μg/L IGF-1 group than the 0 pg/L IGF-1 group ( 1.49±0.18 vs 1.46±0.17 ) ( t =0.335,P =0.743 ),however,the expressing levels of HIF-1α protein( 1049.86±172.54 vs 0.00±0.00) and VEGF mRNA(0.95±0.15 vs 0.35±0.07) and VEGF protein (391.98±56.77 vs 214.36±37.15)were raised in the 50 μg/L IGF-I group compared with 0 μg/L IGF-1 group (t=16.098,9.935,6.928,P<0.05).After the HIF-1α-specific shRNA was transferred into cultured RPE cells,the expressions of both HIF-1α mRNA and its protein significantly decreased in RPE cells under 50 μg/L IGF-1 concentration

  2. Stable shRNA Silencing of Lactate Dehydrogenase A (LDHA) in Human MDA-MB-231 Breast Cancer Cells Fails to Alter Lactic Acid Production, Glycolytic Activity, ATP or Survival.

    Science.gov (United States)

    Mack, Nzinga; Mazzio, Elizabeth A; Bauer, David; Flores-Rozas, Hernan; Soliman, Karam F A

    2017-03-01

    In the US, African Americans have a high death rate from triple-negative breast cancer (TNBC), characterized by lack of hormone receptors (ER, PR, HER2/ERRB2) which are otherwise valuable targets of chemotherapy. There is a need to identify novel targets that negatively impact TNBC tumorigenesis. TNBCs release an abundance of lactic acid, under normoxic, hypoxic and hyperoxic conditions; this referred to as the Warburg effect. Accumulated lactic acid sustains peri-cellular acidity which propels metastatic invasion and malignant aggressive transformation. The source of lactic acid is believed to be via conversion of pyruvate by lactate dehydrogenase (LDH) in the last step of glycolysis, with most studies focusing on the LDHA isoform. In this study, LDHA was silenced using long-term MISSION® shRNA lentivirus in human breast cancer MDA-MB-231 cells. Down-regulation of LDHA transcription and protein expression was confirmed by western blot, immunocytochemistry and qPCR. A number of parameters were measured in fully viable vector controls versus knock-down (KD) clones, including levels of lactic acid produced, glucose consumed, ATP and basic metabolic rates. The data show that lentivirus V-165 generated a knock-down clone most effective in reducing both gene and protein levels to less than 1% of vector controls. Stable KD showed absolutely no changes in cell viability, lactic acid production, ATP, glucose consumption or basic metabolic rate. Given the complete absence of impact on any observed parameter by LDH-A KD and this being somewhat contrary to findings in the literature, further analysis was required to determine why. Whole-transcriptome analytic profile on MDA-MB-231 for LDH subtypes using Agilent Human Genome 4×44k microarrays, where the data show the following component breakdown. Transcripts: 30.47 % LDHA, 69.36% LDHB, 0.12% LDHC and 0.05% LDHD. These findings underscore the importance of alternative isoforms of LDH in cancer cells to produce lactic acid

  3. Inhibition of Bcl-2 expression by a novel tumor-specific RNA interference system increases chemosensitivity to 5-fluorouracil in Hela cells

    Institute of Scientific and Technical Information of China (English)

    Sheng-lin HUANG; Yi WU; Hai YU; Ping ZHANG; Xing-qian ZHANG; Lei YING; Han-fang ZHAO

    2006-01-01

    Aim: RNA interference (RNAi) has been proposed as a potential treatment for cancer, but the lack of cellular targets limits its use in cancer gene therapy. No current technology has achieved direct tumor-specific gene silencing using RNAi.In the present study we attempt to develop a tumor-specific RNAi system using the human telomerase reverse transcriptase (hTERT) promoter; furthermore, we analyzed its inhibitive effect on Bcl-2 expression. Methods: The vectors containing a small hairpin RNA (shRNA) to target exogenous reporters [firefly luciferase and enhanced green fluorescent protein (EGFP)] and endogenous gene (Bcl-2)were constructed. Luciferase expression was determined by dual luciferase assay.Reverse transcription-polymerase chain reaction (RT-PCR), fluorescence microscopy and fluorescence-activated cell sorting (FACS) were used to measure EGFP expression. Inhibition of Bcl-2 was evaluated by RT-PCR and Western blotting.Cell proliferation and viability were measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. FACS was used to analyze the cell cycle distribution profile. Results: We showed that with the hTERT promoter directly driving shRNA transcription, expression of the exogenous reporters (LUC and EGFP) in tumor cells, but not normal cells, was specifically inhibited in vitro. The hTERT promoter-driven shRNA also depressed the expression of Bcl-2. Inhibition of Bcl-2 did not affect cell proliferation, but increased the chemosensitivity of HeLa cells to 5-fluorouracil. Conclusion: The present study describes an efficient RNAi system for gene silencing that is specific to tumor cells using the hTERT promoter. Suppression of Bcl-2 by using this system sensitized HeLa cells to 5-fluorouracil. This system may be useful for RNAi therapy.

  4. Specificity of specific language impairment

    NARCIS (Netherlands)

    GoorhuisBrouwer, SM; WijnbergWilliams, BJ

    1996-01-01

    In children with specific language impairment (SLI) their problems are supposed to be specifically restricted to language. However, both on a theoretical basis as well as on a practical basis it is often difficult to make a sharp distinction between specific and nonspecific language disorders. In a

  5. Construction of shRNA eukaryotic expression vectors targeting PERK gene and effect of PERK gene knockdown on apoptosis in endoplasmic reticulum stress-induced L02 hepatocytes%靶向PERK基因的shRNA真核表达载体的构建及对内质网应激状态下L02肝细胞凋亡的影响

    Institute of Scientific and Technical Information of China (English)

    曹洁; 杨朝霞; 沈薇; 姚隆

    2011-01-01

    AIM: To construct short hairpin RNA ( shRNA ) eukaryotic expression vectors targeting the gene of RNA - dependent protein kinase ( PKR ) - like endoplasmic reticulum kinase ( PERK ), and to observe the effect of PERK gene knockdown on apoptosis of human normal hepatic L02 cells treated with thapsigargin. METHODS: Three shRNA expression vectors targeting PERK gene, named PERK1 - shRNA, PERK2 - shRNA and PERKS - shRNA, and one non -homologous negative control expression vector ( HK - shRNA ) were constructed based on the nucleotide sequence of PERK and the criteria of designing small interfering RNA ( siRNA ), and were identified by enzyme digestion and DNA sequencing analysis. After L02 hepatocytes were transfected with the plasmids, the PERK expression was determined by RT - PCR and Western blotting, and the plasmid with the best inhibitory effect on PERK expression was screened. The cell viability and apoptotic rate of L02 hepatocytes transfected with PERK - shRNA under endoplasmic reticulum stress ( ERS ) were measured by the methods of MTT and flow cytometry,respectively. RESULTS: Four shRNA expression vectors were constructed. The mRNA and protein expression levels of PERK gene decreased significantly in L02 hepatocytes transfected with PERK1 - shRNA, PERK2 - shRNA and PERK3 - shRNA as compared with those in control cells ( P <0. 05 ). The inter-fering effect of PERK1 - shRNA on PERK gene expression was the best. PERK knockdown by PERK1 - shRNA increased the viability and inhibited the apoptosis of L02 cells under ERS. CONCLUSION: The shRNA expression vectors targeting PERK gene are constructed, and PERK gene knockdown may inhibit apoptosis in L02 hepatocytes under ERS.%目的:构建靶向RNA依赖的蛋白激酶样内质网激酶(PERK)基因的短发夹状RNA(shRNA)真核表达载体,观察其对内质网应激(ERS)状态下人正常肝细胞株L02凋亡的影响.方法:根据PERK基因核苷酸序列,按照小干扰RNA(siRNA)靶序列的设计原则,设

  6. The Comparison of Inhibition Effect to HSV-2 of UL29shRNA Expression Plasmid and ACV%UL29shRNA表达质粒与ACV对HSV-2抑制效果的比较

    Institute of Scientific and Technical Information of China (English)

    仵立佳; 吕延成; 兰小凇; 潘晓瑜; 丁娟

    2016-01-01

    目的:通过将筛选的UL29shRNA表达质粒与抗病毒药物阿昔洛韦(ACV)的抗病毒效果及细胞毒性进行比较,探讨RNA干扰在细胞水平上对HSV-2病毒的抑制效果.方法:将筛选的干扰效果最好的表达质粒UL29-shRNA 1641作为RNA干扰组与抗病毒药ACV进行对HSV-2的抑制效果的比较研究,分为RNAi组、ACV组和RNAi+ACV组.采用实时荧光定量PCR检测UL29mRNA的相对表达量,Karber法检测细胞上清液中病毒滴度的变化、蛋白印迹法(Western blot)检测ICP8的相对表达量,对RNAi与ACV抑制效果进行比较,通过WST-1(Water-Soluble Tetrazolium-1)法对质粒转染复合物(质粒+转染试剂)和ACV的细胞毒性进行比较.结果:RT-PCR检测三组mRNA的相对表达水平,其中RNAi组UL29基因相对表达量高于ACV组(P<0.05);RNAi+ACV组UL29基因相对表达量明显低于RNAi组和ACV组(P<0.05).Karber法检测三组细胞上清液病毒滴度表明,RNAi组病毒滴度高于ACV组(P<0.05),RNAi+ACV组的病变病毒滴度明显低于RNAi组和ACV组(P<0.05).Western blot检测ICP8相对表达量,ACV组的ICP8(UL29编码的单链DNA结合蛋白,主要作用是在HSV-2 DNA复制过程中与复制叉产生了单链DNA结合,防止其重新配对形成dsDNA或被核酸酶降解)相对表达量比RNAi组低(P<0.05).RNAi+ACV组ICP8相对表达量降低最为明显,与ACV组和RNAi组相比有显著差异(P<0.05).WST-1法对转染复合物与ACV的细胞毒性进行比较,其中RNAi中质粒和转染试剂对细胞的毒性明显小于ACV.结论:在RNAi与ACV对HSV-2抑制效果的比较中,ACV要好于RNAi,两者联合使用的抑制效果好于单独使用RNAi或ACV.在抑制效果都较好的情况下比较,RNAi中使用的质粒与转染试剂对细胞的影响比ACV小.

  7. shRNA沉默ROCK1和ROCK2表达对缺氧诱导的大鼠心肌细胞凋亡的影响%Effect of ROCK1 and ROCK2 silencing by shRNA on hypoxia-induced apoptosis of rat cardiomyocytes

    Institute of Scientific and Technical Information of China (English)

    孙国芳; 丁浩; 李菊香; 洪葵; 董家龙; 吴清华; 程晓曙

    2011-01-01

    ) and R0CK2 on apoptosis induced by hypoxia in rat cardiomyocytes. METHODS: Rat cardiomyocytes were cultured primarily and identified using an antibody targeting a. - actin of striated muscle. R0CK1 - shRNA and R0CK2 - shRNA were transiently transfected into the cells by liposome. After 48 h, these cells were subject to hypoxia for 6 h. The cells were divided into 5 groups: blank control group, hypoxia group, hypoxia + negative control shRNA group, hypoxia + R0CK1 - shRNA group and hypoxia + R0CK2 - shRNA group. The beating frequency and rhythm of the cardiomyocytes were assessed by microscopy. The activity of lactate dehydrogenase ( LDH ) in the cell culture supernatants was detected by automatic biochem-ical analyzer. The cell survival rate was analyzed by the method of MTT. The cell apoptotic rate was assessed by flow cy-tometry. Western blotting was used to determine the expression of R0CK1, R0CK2, caspase - 3 and p - PDK. RESULTS : The primary culture of the cardiomyocytes was successful. Western blotting results showed that the transfection of R0CK1 - shRNA or R0CK2 - shRNA decreased the expression of R0CK1 or R0CK2 in the cardiomyocytes. Hypoxia slowed down the beat frequency of the cardiomyocytes, also made the rhythm disorder. Hypoxia increased the release of LDH and decreased the cell survival rate. Flow cytometry results showed that hypoxia increased the cell apoptotic rate. Hypoxia increased the expression of caspase - 3 and decreased the expression of p - PDK. Transfection of R0CK1 - shRNA and R0CK2 - shRNA into the cardiomyocytes reduced all the effects of hypoxia mentioned above. CONCLUSION: Down - regulation of R0CK1 and R0CK2 expression suppresses the apoptosis of rat cardiomyocytes induced by hypoxia. The mechanism is associated with the inhibition of caspase - 3 activation and the up - regulation of p - PDK expression.

  8. Specific Dyslexia.

    Science.gov (United States)

    Naidoo, Sandhya

    The investigation reported in this volume attempts to clarify some issues relating to the existence, nature, and causes of specific dyslexia. Based on an extended study of 98 boys of at least average intelligence with severe reading and spelling problems, the report provides detailed data relating to their developmental and perinatal histories,…

  9. Heritable and lineage-specific gene knockdown in zebrafish embryo.

    Directory of Open Access Journals (Sweden)

    Mei Dong

    Full Text Available BACKGROUND: Reduced expression of developmentally important genes and tumor suppressors due to haploinsufficiency or epigenetic suppression has been shown to contribute to the pathogenesis of various malignancies. However, methodology that allows spatio-temporally knockdown of gene expression in various model organisms such as zebrafish has not been well established, which largely limits the potential of zebrafish as a vertebrate model of human malignant disorders. PRINCIPAL FINDING: Here, we report that multiple copies of small hairpin RNA (shRNA are expressed from a single transcript that mimics the natural microRNA-30e precursor (mir-shRNA. The mir-shRNA, when microinjected into zebrafish embryos, induced an efficient knockdown of two developmentally essential genes chordin and alpha-catenin in a dose-controllable fashion. Furthermore, we designed a novel cassette vector to simultaneously express an intronic mir-shRNA and a chimeric red fluorescent protein driven by lineage-specific promoter, which efficiently reduced the expression of a chromosomally integrated reporter gene and an endogenously expressed gata-1 gene in the developing erythroid progenitors and hemangioblasts, respectively. SIGNIFICANCE: This methodology provides an invaluable tool to knockdown developmental important genes in a tissue-specific manner or to establish animal models, in which the gene dosage is critically important in the pathogenesis of human disorders. The strategy should be also applicable to other model organisms.

  10. 质粒shRNA体内干扰Klotho基因对窦房结通道基因的影响%Study on the Effect of Klotho Gene Interferred by Plasmid-mediated Short Hairpin RNA (shRNA) on Sinoatrial Node Pacing Channel Gene

    Institute of Scientific and Technical Information of China (English)

    蔡盈盈; 汪汉; 侯言彬; 房晨鹂; 田鹏; 王贵华; 李璐; 邓珏琳

    2013-01-01

    通过质粒shRNA体内干扰,研究Klotho基因与窦房结起搏通道相关基因HCN4及HCN2之间的关系,为病窦综合征的研究提供新思路.取C57BL/6J小鼠20只,分为4组,每组5只,分别为:质粒shRNA 24 h组、质粒shRNA 12 h组、生理盐水24 h组、生理盐水12h组.质粒shRNA组经尾静脉注射质粒shRNA 50 μL(1 μg质粒/μL),生理盐水组经尾静脉注射生理盐水50 μL.分别于注射12h及24 h后取窦房结周围组织,行RT-PCR检测各组小鼠的Klotho、HCN2、HCN4基因的mRNA水平.RT-PCR结果显示:与生理盐水12h组比较,shRNA 12 h组的klotho、HCN4和HCN2的mRNA表达量明显降低,均有统计学差异(P<0.05).以上结果提示,小鼠Klotho基因和窦房结起搏基因可能存在一定关系.%The study was aimed to assess the effect of Klotho gene and sinoatrial node pacing channel gene (HCN4and HCN2) for studying sick sinus syndrome,with Klotho gene under the interference of Plasmid-mediated short hairpin RNA.Twenty-five C57BL/6J mice were divided into four groups,i.e,plasmid shRNA 24h group,plasmid shRNA 12h group,sodium chloride 24h group and sodium chloride 12h group.Plasmid shRNA 50μL (1μg/μL) and sodium chloride 50μl were respectively injected according to mice vena caudalis into those in plasmid shRNA group and sodium chloride group.After 12h or 24h respectively,all mice were executed and their sinoatrial node tissues were cut.The mRNA of Klotho,HCN4 and HCN2 gene were detected by RT-PCR.The results of RT-PCR showed that Klotho,HCN4 and HCN2 mRNA levels were lower compared with those in sodium chloride 12h group after 12h interference interval.The results indicated that there might be the a certain relationship between Klotho gene and sinoatrial node pacing channel gene.

  11. CD147-shRNA对肝癌细胞CD147表达的抑制作用%The inhibitory effect of CD147-shRNA on expression of CD147 of hepatoma cell line SMMC-7721 and HepG2

    Institute of Scientific and Technical Information of China (English)

    宫路路; 林瑞新; 李晓萌; 张爱丽; 文海荣; 房学东

    2010-01-01

    目的 构建CD147-shRNA重组质粒并检测其对肝癌细胞SMMC-7721和HepG2内源性CD147表达的抑制作用.方法 根据Genebank中CD147序列设计3条特征性靶序列,合成的核苷酸序列连接到线性化pBS/U6载体,采用酶切法及测序鉴定重组质粒的正确性;将重组质粒转染至人肝癌细胞SMMC-7721和HepG2中,采用Western blot法和免疫荧光法观察该重组质粒对CD147内源性表达的抑制作用.结果 酶切鉴定和测序证实成功构建pBS/U6/CD147-shRNA,且对人肝癌细胞SMMC7721和HepG2 CD147表达有抑制作用,而以pBS/U6/CD147-shRNA3抑制作用最为显著.结论 构建pBS/U6/CD147-shRNA成功,并筛选出基因抑制效果最佳的pBS/U6/CD147-shRNA3,为进一步实验打下了基础.

  12. HRT Specification

    CERN Document Server

    Möller, M

    1996-01-01

    In the context of the AIS Project (Advanced Informatics Systems for administration and management) a study has been conducted that resulted in the definition of a high level information systems model. Thirteen proposed systems were defined for detailed analysis. The Finance, Foundation, Human Resources, Logistics and Purchasing areas have been studied in detail. These studies have lead to the purchase and implementation of the ORIAC and SIRIAC packages, the Foundation database, the Oracle HR package, the Triton package and EDH and BHT. This specification describes the Human Resources Toolkit (HRT) intended to be used for accessing data in the HR and Foundation systems. This toolkit should help the divisions carry out their Human Resource management, planning and follow-up. It will have extensive report generation capabilities and offer a variety of standard graphs. It should have an easy-to-use graphical user interface and run on the CERN standard desktop platforms.

  13. Specific phobias.

    Science.gov (United States)

    Hamm, Alfons O

    2009-09-01

    Exposure based treatments in which patients are systematically confronted with their feared objects of situations are highly effective in the treatment of specific phobias and produce stable improvement both in reported fear and behavioral avoidance. Exposure in reality is more effective in most cases than exposure in sensu. For situations that are difficult to realize, exposure in virtual environments has become increasingly valuable. Exposure in vivo is clearly superior to pharmacotherapy, although cognitive enhancers have been successfully used recently to increase the effect of exposure therapy. The induction of relaxation is not a necessary precondition for exposure therapy. Rather the current mechanisms of change focus on extinction learning as being the central mechanism both on a cognitive level namely that the feared object is no longer associated with severely threatening consequence but also on an affective level, meaning that feared cue is no longer capable to activate the fear circuit in the brain. Accordingly future diagnostic categorizations of phobic disorders in the DSM-V should rather focus on the pattern of the fear response that needs to be changed than on the eliciting cues or situations that are avoided.

  14. Effect of lentivirus-mediated shRNA targeting collagen type Ⅰ gene on biological behaviors of rat mesangjal cells%慢病毒介导Ⅰ型胶原短发夹RNA转染对大鼠系膜细胞生长行为的影响

    Institute of Scientific and Technical Information of China (English)

    单毅; 周春华; 赵海丹

    2010-01-01

    Objective By observing the change of proliferation, apoptosis and cell cycle about RMC infec-ted by lentivirus-mediated COL I shRNA, we explore the physiological impact on the target cells by RNAi medicine .Methods We observed the infected RMC by Infection enhancer( control group),lentivirus vector( pSC-GFP group) and lentivirus-mediated COL Ⅰ shRNA (pSC-GFP/COL Ⅰ group) ,cell proliferation was measured with MTT method,apoptosis was measured with Annexin V/PE method,cell cycle was measured with flow cytometry and the impact on the above-mentioned ability of cells infected by lentiviral expression vector was observed. Results Both pSC-GFP/COL Ⅰ group and pSC-GFP group can significant inhibit cell proliferation. Suppression ability of pSC-GFP group was significantly higher than pSC-GFP group. 24 hours,48 hours and 72 hours after infection showed inhibition efficiency of 22.52% , 28.57% , 33.33% , respectively. The result of apoptosis show that apoptosis was induced to a certain degree after lentivirus-mediated shRNA infected RMC, but RNAi didnt induce further aggravation. The result of cell cycle experiment showed that lentivirus induces G2/M cell cycle arres. To 72 houres after RMC infected by lentiviral expression vector, increasingly prominent showed that COL Ⅰ shRNA induced S cell cycle arrest. Conclusions Af-ter transfection of rat mesangial cells, COL Ⅰ shRNA affects cell proliferation at a certain degree. But on the whole,its a safe way to lay the foundation for further animal experiment and drug development.%目的 通过观察慢病毒介导的Ⅰ型胶原((COL Ⅰ)短发夹(sh)RNA感染大鼠系膜细胞后其增殖、凋亡及细胞周期的变化,探索未来应用于动物实验,甚至临床时RNA干涉(RNAi)药物对靶细胞生长行为的影响.方法 利用感染增强剂(对照组)、空慢病毒载体及感染增强剂(pSC-GFP组)、COL Ⅰ shRNA慢病毒表达载体及感染增强剂(pSC-GFP/COL Ⅰ组)分别感染大鼠系膜细胞,用

  15. Random shRNA Library Screen for shRNAs Targeting HIV-1 LTR Related Host Factors with TK Suicide Gene%shRNA随机文库结合TK自杀基因筛选靶向HIV-1 LTR相关宿主因子

    Institute of Scientific and Technical Information of China (English)

    李建彬; 方祥; 童贻刚; 米志强; 安小平; 谭莉; 陈斌; 王晓娜; 范华昊; 张文慧; 张博

    2012-01-01

    构建shRNA随机文库与HIV-1 LTR启动胸苷激酶基因(TK基因)稳定表达的稳定细胞系HEK293/TK,将两者结合起来,筛选靶向HIV-1 LTR相关宿主因子.方法:通过化学合成含有19个随机脱氧核苷酸的发夹结构,将其退火补平后与合成的接头Linker连接进行PCR反应,将PCR产物酶切后置于慢病毒载体pLenti-U6启动子下游由此构建shRNA随机文库;利用重叠PCR将HIV-1 LTR片段和TK基因连接起来,连接产物经酶切后与pcDNA3.1载体连接;将连接正确的质粒转染HEK293细胞同时用G418加压筛选获得稳定细胞系HEK293/TK;将所获得的文库质粒包装成慢病毒后侵染所构建的HEK293/TK细胞系,通过加入药物GCV进行加压筛选获得存活细胞.结果:成功筛选到加药后存活下来的细胞,抽提细胞基因组,采用巢式PCR扩增目的干扰序列并用Western blot对干扰序列进行验证,鉴定获得一个克隆所表达的shRNA能对TK基因的表达起到抑制作用,通过测序分析获得其干扰序列,该序列很有可能针对HIV-1 LTR某宿主相关因子.结论:成功构建了一种筛选HIV-1 LTR相关宿主因子的方法,筛选所得序列可以定位到具体相关宿主因子,为靶向筛选抗HIV-1药物提供了重要手段.%To screen HIV-1 LTR related host factors by constructing random RNAi library based on the lentiviral vector and HIV-1 LTR reporter cell line in which TK suicide gene can express stably. Methods: Oligo deoxynucleotides were chemically synthesized which formed hairpin structure that contain 19 random bases and were then fused with a linker sequence. The joined product was amplified by PCR and the PCR product was cloned downstream of U6 promoter of a lentiviral vector pLenti-U6 to prepare random shRNA Library. HIV-1 LTR and HSV-TK gene were amplified by PCR and then joined by overlapping PCR. The joined product was cloned into pcDNA3. 1 expression vector. The recombinant plasmid obtained was transfected into HEK293

  16. Primate-specific Melanoma Antigen-A11 Regulates Isoform-specific Human Progesterone Receptor-B Transactivation*

    Science.gov (United States)

    Su, Shifeng; Blackwelder, Amanda J.; Grossman, Gail; Minges, John T.; Yuan, Lingwen; Young, Steven L.; Wilson, Elizabeth M.

    2012-01-01

    Progesterone acting through the progesterone receptor (PR) and its coregulators prepares the human endometrium for receptivity to embryo implantation and maintains pregnancy. The menstrual cycle-dependent expression of melanoma antigen-A11 (MAGE-11) in the mid-secretory human endometrium suggested a novel function in human PR signaling. Here we show that MAGE-11 is an isoform-specific coregulator responsible for the greater transcriptional activity of human PR-B relative to PR-A. PR was recruited to progesterone response regions of progesterone-regulated FK506-binding protein 5 (FKBP5) immunophilin and small Ras family G protein cell growth inhibitor RASD1 genes. Expression of MAGE-11 lentivirus shRNA in human endometrial Ishikawa cells expressing PR-B showed that MAGE-11 is required for isoform-specific PR-B up-regulation of FKBP5. In contrast, MAGE-11 was not required for progesterone up-regulation of RASD1 in endometrial cells expressing the PR-A/B heterodimer. Target gene specificity of PR-B depended on the synergistic actions of MAGE-11 and p300 mediated by the unique PR-B NH2-terminal 110LLXXVLXXLL119 motif that interacts with the MAGE-11 F-box region in a phosphorylation- and ubiquitinylation-dependent manner. A progesterone-dependent mechanism is proposed in which MAGE-11 and p300 increase PR-B up-regulation of the FKBP5 gene. MAGE-11 down-regulates PR-B, similar to the effects of progesterone, and interacts with FKBP5 to stabilize a complex with PR-B. We conclude that the coregulator function of MAGE-11 extends to isoform-specific regulation of PR-B during the cyclic development of the human endometrium. PMID:22891251

  17. Therapy for dominant inherited diseases by allele-specific RNA interference: successes and pitfalls.

    Science.gov (United States)

    Trochet, Delphine; Prudhon, Bernard; Vassilopoulos, Stéphane; Bitoun, Marc

    2015-01-01

    RNA interference (RNAi) is a conserved mechanism for post-transcriptional gene silencing mediated by messenger RNA (mRNA) degradation. RNAi is commonly induced by synthetic siRNA or shRNA which recognizes the targeted mRNA by base pairing and leads to target-mRNA degradation. RNAi may discriminate between two sequences only differing by one nucleotide conferring a high specificity of RNAi for its target mRNA. This property was used to develop a particular therapeutic strategy called "allele-specific-RNA interference" devoted to silence the mutated allele of genes causing dominant inherited diseases without affecting the normal allele. Therapeutic benefit was now demonstrated in cells from patients and animal models, and promising results of the first phase Ib clinical trial using siRNA-based allele-specific therapy were reported in Pachyonychia Congenita, an inherited skin disorder due to dominant mutations in the Keratin 6 gene. Our purpose is to review the successes of this strategy aiming to treat dominant inherited diseases and to highlight the pitfalls to avoid.

  18. 靶向CD40的shRNA干扰抗大鼠异体肢体移植急性排斥反应的实验研究%Resistance to acute rejection by shRNA interference from CD40 costimulatory molecule in limb allo-transplantation in rats

    Institute of Scientific and Technical Information of China (English)

    张震宇; 刘伟; 毕郑钢; 董清平

    2008-01-01

    目的 探讨靶向CD40的RNA干扰对大鼠异体肢体移植急性排斥反应的影响. 方法以纯系SD大鼠为供体,纯系Wistar大鼠为受体,行同种异体右后肢移植.27只大鼠肢体移植后随机分为三组,A组:注射入梭华.Sofast.siCD40-2/pSilencer载体复合物600 μL;B组:注射Sofast-pSilencer4.1-CMV neo空载体复合物600 μL;C组:注射生理盐水600μL,以上均通过阴茎背静脉注射.观察移植物排斥反应征象及存活情况,并于第7天对产生免疫耐受大鼠进行混合淋巴细胞反应,同时进行组织学检查. 结果与B、C组相比,A组移植物发生排斥反应的时间及存活时间均显著延长,差异有统计学意义(P<0.01)(>13 d),未见排斥反应征象;B、C组均于术后近期发牛排斥反应.A组大鼠对供体的淋巴细胞呈现低反应性,移植的供体同系大鼠的肢体得以存活. 结论术后不应用免疫抑制剂的情况下,靶向CD40的shRNA干扰可以抗大鼠异体肢体移植急性排斥反应.%Objective To study the resistance to acute reaction in composite tissue allotransplan-tation by shRNA interference from CD40 costimulatory molecule in rat lymphocytes. Methods The in-bred SD rats were chosen as donors and inbred Wistar rats as recipients. Twenty-seven recipients were divided into 3 groups randomly after allogeneic leg transplantation. On the day after transplantation, rats in group A were injected with 600 μL of Sofast-siCD40-2/pSilencer, rats in group B received 600 μL of Sofast-pSilencer 4.1-CMV neo, and rats in group C received injection of 600 μL of sodium chloride. The survival and rejection to grafts were observed. The rats which survived 7 days after operation were given the test of mixed lymphocyte reaction (MIR). Results The mean survival time of limb allografts in group A was obviously longer than that in the other two groups( P < 0.01 ) . Rejection to grafts was not observed in group A, but rejection e-merged shortly after

  19. Study on the effect of PDGF-D shRNA on proliferation and invasion of breast cancer SK-BR-3 cells%PDGF-D 稳定沉默后对乳腺癌细胞 SK-BR-3生物学功能的影响

    Institute of Scientific and Technical Information of China (English)

    李红昌; 冯雯; 陈亚峰; 梅怡; 蔡含; 蒋一鸣; 陈腾; 殷佩浩; 奉典旭

    2016-01-01

    目的:通过慢病毒载体沉默乳腺癌细胞系 SK - BR -3中血小板衍生生长因子 D(PDGF - D)基因的表达,探讨 PDGF - D 基因沉默后对乳腺癌细胞增殖、凋亡及迁移的调控作用。方法应用 RT - PCR 及蛋白质印迹方法检测5种乳腺癌细胞系中 PDGF - D 的表达状况;设计人 PDGF - D 基因的 shRNA 慢病毒载体,在293T 细胞中包装病毒并感染 PDGF - D 高表达的乳腺癌细胞系,运用蛋白质免疫印迹的方法鉴定其对 PDGF - D 基因的沉默效果;PDGF - D基因沉默后通过细胞增殖实验和软琼脂克隆形成实验检测其对细胞增殖能力的影响,并应用流式细胞技术检测细胞凋亡;运用 Transwell 迁移实验检测细胞的体外侵袭迁移能力。结果 PDGF - D 在5种细胞系中存在差异性表达,其中具有低转移潜能的 HT -29和 SK - BR -3细胞与具有高转移潜能的 LOVO、RKO 和 SW620细胞相比,具有更高的 PDGF -D 表达水平( t =3.880,P <0.05)。选取低转移细胞株中 PDGF - D 表达相对较高的 SK - BR -3作为研究对象,构建PDGF - D shRNA 慢病毒载体沉默 SK - BR -3细胞中 PDGF - D 的表达,蛋白质免疫印迹结果提示,PDGF - D 稳定沉默的 SK - BR -3细胞系构建成功;在稳定沉默 PDGF - D 基因后,MTT 细胞增殖实验和软琼脂克隆形成实验表明 SK - BR-3细胞增殖能力增强( F =75.23,P <0.001);流式细胞技术检测显示 PDGF - D 基因沉默后可抑制 SK - BR -3细胞凋亡( F =84.44,P <0.001);细胞小室实验结果显示,PDGF - D 沉默后 SK - BR -3迁移能力增强( F =155.9,P <0.001)。结论 PDGF - D 基因可能与乳腺癌的细胞增殖、凋亡与转移有关,进而参与了乳腺癌的发生、发展。%Objective The shRNA lentiviral vector was constructed to silence PDGF - D expression in BrCa cells,in order to explore the role of PDGF - D in proliferation,apoptosis and migration

  20. Myositis specific autoantibodies : specificity and clinical applications

    NARCIS (Netherlands)

    Hengstman, G.J.D.

    2005-01-01

    The sera of about half of the patients with myositis contain autoantibodies that are specific for this group of diseases compared to other inflammatory connective tissue disorders. In a recent study we showed that these myositis specific autoantibodies (MSAs) are also specific for myositis as compar

  1. Myositis specific autoantibodies : specificity and clinical applications

    NARCIS (Netherlands)

    Hengstman, G.J.D.

    2005-01-01

    The sera of about half of the patients with myositis contain autoantibodies that are specific for this group of diseases compared to other inflammatory connective tissue disorders. In a recent study we showed that these myositis specific autoantibodies (MSAs) are also specific for myositis as compar

  2. Expression of RNA interference triggers from an oncolytic herpes simplex virus results in specific silencing in tumour cells in vitro and tumours in vivo

    Directory of Open Access Journals (Sweden)

    Anesti Anna-Maria

    2010-09-01

    Full Text Available Abstract Background Delivery of small interfering RNA (siRNA to tumours remains a major obstacle for the development of RNA interference (RNAi-based therapeutics. Following the promising pre-clinical and clinical results with the oncolytic herpes simplex virus (HSV OncoVEXGM-CSF, we aimed to express RNAi triggers from oncolytic HSV, which although has the potential to improve treatment by silencing tumour-related genes, was not considered possible due to the highly oncolytic properties of HSV. Methods To evaluate RNAi-mediated silencing from an oncolytic HSV backbone, we developed novel replicating HSV vectors expressing short-hairpin RNA (shRNA or artificial microRNA (miRNA against the reporter genes green fluorescent protein (eGFP and β-galactosidase (lacZ. These vectors were tested in non-tumour cell lines in vitro and tumour cells that are moderately susceptible to HSV infection both in vitro and in mice xenografts in vivo. Silencing was assessed at the protein level by fluorescent microscopy, x-gal staining, enzyme activity assay, and western blotting. Results Our results demonstrate that it is possible to express shRNA and artificial miRNA from an oncolytic HSV backbone, which had not been previously investigated. Furthermore, oncolytic HSV-mediated delivery of RNAi triggers resulted in effective and specific silencing of targeted genes in tumour cells in vitro and tumours in vivo, with the viruses expressing artificial miRNA being comprehensibly more effective. Conclusions This preliminary data provide the first demonstration of oncolytic HSV-mediated expression of shRNA or artificial miRNA and silencing of targeted genes in tumour cells in vitro and in vivo. The vectors developed in this study are being adapted to silence tumour-related genes in an ongoing study that aims to improve the effectiveness of oncolytic HSV treatment in tumours that are moderately susceptible to HSV infection and thus, potentially improve response rates seen

  3. Effects of c-fos Down-regulation via shRNA on P-gp-mediated Multidrug Resistance in Human Breast Cancer MCF-7/ADR Cells%shRNA抑制c-fos表达对P-gp介导的乳腺癌多药耐药的影响

    Institute of Scientific and Technical Information of China (English)

    师锐赞; 胡晓玲; 范彦英

    2012-01-01

    多药耐药(multidrug resistance,MDR)是导致化疗失败的重要原因,多药耐药基因(multidrug resistance gene,mdr1)产物P-糖蛋白(P-glycoprotein,P-gp)过表达是最主要的耐药机制.原癌基因c-fos在肿瘤MDR中的作用渐受重视.主要选用人乳腺癌敏感株MCF-7和阿霉素(adriamycin,ADR)筛选的、mdr1/P-gp高表达的耐药株MCF-7/ADR,探讨c-fos在P-gp介导的乳腺癌MDR中的作用.相对于MCF-7,c-fos在MCF-7/ADR高表达.采用shRNA法下调c-fos表达后,MCF-7/ADR对ADR的敏感性大大增强,且mdr1/P-gp表达减少、P-gp外排功能降低.c-fos表达下调可逆转对P-gp介导的乳腺癌MDR的实验结果,为c-fos成为逆转肿瘤耐药诊断和治疗的新靶标,对实现耐药乳腺癌的分子靶向治疗提供了理论基础.%Multidrug resistance (MDR) is the main reason of chemotherapy failure. The overexpression of P-glycoprotein (P-gp) , encoded by the multidrug resistance (mdrl) gene, is thought to be the major cause of MDR phenotype. Since much attention has been paid to the role of proto-oncogene c-fos in MDR, adriamycin (ADR)-selected resistant breast cancer cells (MCF-7/ADR) with mdrl/P-gp overexpression and parental drug-sensitive cells ( MCF-7) were chosen to analyze the role of c-fos in P-gp-mediated MDR. Elevated c-fos expression is observed in MCF-7/ADR compared to MCF-7 cells. Down-regulation of c-fos expression via shRNA resulted in sensitization of MCF-7/ADR cells to ADR and decreased the expression of mdrl/P-gp and efflux function of P-gp. Based on these results, c-fos may represent a potential molecular target for resistant cancer therapy, and suppressing c-fos gene expression may therefore be an effective means for targeted molecular therapy.

  4. Expression of aquaporin-1 in rat pleural mesothelial cells and its specific inhibition by RNA interference in vitro

    Institute of Scientific and Technical Information of China (English)

    ZHANG Wei; XIE Can-mao; LI Zhi-ping

    2007-01-01

    Background The discovery of water channel aquaporins(AQPs)has greatly expanded the understanding of the regulation of the water permeability of biological membranes.Aquaporin-1(AQP1)may be involved in fluid transport in numerous pathological conditions.The objective of the present study was to examine whether AQP1 is present in cultured rat pleural mesothelial cells(PMCs)and to investigate the specific inhibitory effect of RNA interference(RNAi) on AQP1 expression in PMCs,which may provide a new method for the further studies on the relation between expression of AQP1 in PMCs and pleural fluid removal in vivo.Methods PMCs were isolated and cultured from rat pleura.The expression of AQP1 in PMCs was confirmed by immunocytochemical staining and reverse transcriptase-polymerase chain reaction(RT-PCR).Two eukaryotic expression plasmid vectors of short hairpin RNA(shRNA)specific for the AQP1 gene of rat sapien were designed and constructed.The recombinant plasmid vectors were transfected into cultured rat PMCs by cation liposomes.Flow cytometry was used to screen the most effective shRNA at 48 hours after transfection.The expressions of AQP1 mRNA and protein were detected by RT-PCR and Western blotting method at 48 hours after transfection.Results RT-PCR and immunostaining revealed that AQP1 mRNA and protein were present in cultured rat PMCs.Two effective eukaryotic expression plasmid vectors of shRNA specific for the AQP1 gene were constructed successfully.The levels of the expression of AQP1 were inhibited by 83.45%,90.93%,respectively,at mRNA level and 41.24%,67.60%,respectively at protein level by two recombinant plasmids at 48 hours after transfection.The expression of AQP1 in PMCs transfected with plasmid was significantly lower than that of the cells transfected with the control plasmid HK and that of the untransfected cells(P<0.01).There was no significant difference in AQP1 expression between the control group and the group transfected with AQP1 nonspecific sh

  5. Urine specific gravity test

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003587.htm Urine specific gravity test To use the sharing features on this page, please enable JavaScript. Urine specific gravity is a laboratory test that shows the concentration ...

  6. Phobia - simple/specific

    Science.gov (United States)

    Anxiety disorder - phobia ... Specific phobias are a type of anxiety disorder in which a person may feel extremely anxious or has a ... when exposed to the object of fear. Specific phobias are a common mental disorder. Common phobias include ...

  7. 双基因shRNA干扰乙酰肝素酶在卵巢癌细胞中表达的实验研究%Inhibition of heparanase gene expression by double shRNA expressing vector in ovarian carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    刘玉; 辛晓燕; 毛敬; 张潍

    2006-01-01

    目的:采用编码2条短发卡式RNA(shRNA)的抗乙酰肝素酶(heparanase,Hpa)特异性载体干扰卵巢癌细胞中Hpa的表达,观察其对肿瘤生长转移的抑制作用.方法:以脂质体介导的方法将自行设计和构建的抗Hpa短发卡式RNA(Hpa-shRNA)基因真核表达载体稳定转染人卵巢癌细胞SKOV3;流式细胞仪鉴定转染效率,免疫组化和实时定量RT-PCR检测转染前后卵巢癌细胞中Hpa蛋白和mRNA表达,Boyden小室观测细胞增殖和侵袭能力的变化,透射电镜观察卵巢癌细胞超微结构改变.结果:Hpa mRNA表达在SKOVa-Hpa(转染Hpa)明显升高(P<0.01),SKOV3-Hpa shRNA(转染Hpash RNA)显著降低(P<0.01),SKOV3 DZ-shRNA(转染非特异性sh RNA片段)和SKOV3无显著性差异(P>0.05).电镜下,SKOV3-Hpa组Hpa蛋白表达最为明显,SKOV3-Hpa shRNA Hpa蛋白则最低,各组间比较差别有显著性意义(P<0.01).SKOV3-Hpa细胞器发达,细胞表面绒毛丰富,细胞间连接稀少.SKOV3 Hpa-shRNA以变形为主,细胞表面绒毛少,细胞器不发达,部分见线粒体皱缩,肿胀和空泡样改变.SKOV3-Hpa、SKOV3-DZ-shRNA和SKOV3-Hpa-shRNA 3种细胞穿过人工基底膜的细胞数分别为82.3±9.16,36.1±8.73和18.6±10.5,各组间比较有显著性差异(P<0.01).结论:双基因Hpa特异性shRNA可显著抑制卵巢癌细胞Hpa表达,降低细胞侵袭能力.

  8. The SIFT Code Specification

    Science.gov (United States)

    1983-01-01

    The specification of Software Implemented Fault Tolerance (SIFT) consists of two parts, the specifications of the SIFT models and the specifications of the SIFT PASCAL program which actually implements the SIFT system. The code specifications are the last of a hierarchy of models describing the operation of the SIFT system and are related to the SIFT models as well as the PASCAL program. These Specifications serve to link the SIFT models to the running program. The specifications are very large and detailed and closely follow the form and organization of the PASCAL code. In addition to describing each of the components of the SIFT code, the code specifications describe the assumptions of the upper SIFT models which are required to actually prove that the code will work as specified. These constraints are imposed primarily on the schedule tables.

  9. Experiences with effects specifications

    DEFF Research Database (Denmark)

    2011-01-01

    We describe the effects-specification process from a project that was conducted during the fall 2010 and spring of 2011 in this chapter. The project configured and implemented an electronic patient record system at a maternity ward at a hospital located in a European region. The process comprised...... for measuring effects were designed. The project is analyzed and lessons learned are discussed....... workshops with effects specification with management and end-users and an agile development process including prototypes configured from the effects specifications. We describe the project and the effects-specification process through which effects were related to the system design and instruments...

  10. Screening for Specific Phobias

    Science.gov (United States)

    ... Disorder Specific Phobias Obsessive-Compulsive Disorder (OCD) Posttraumatic Stress Disorder (PTSD) Depression Bipolar Disorder Suicide and Prevention Stress Related Illnesses Myth-Conceptions Find ...

  11. Lidar base specification

    Science.gov (United States)

    Heidemann, Hans Karl.

    2012-01-01

    In late 2009, a $14.3 million allocation from the “American Recovery and Reinvestment Act” for new light detection and ranging (lidar) elevation data prompted the U.S. Geological Survey (USGS) National Geospatial Program (NGP) to develop a common base specification for all lidar data acquired for The National Map. Released as a draft in 2010 and formally published in 2012, the USGS–NGP “Lidar Base Specification Version 1.0” (now Lidar Base Specification) was quickly embraced as the foundation for numerous state, county, and foreign country lidar specifications.

  12. Improved technical specifications

    Energy Technology Data Exchange (ETDEWEB)

    Hoffman, D.R.

    1994-12-31

    Improved technical specifications for nuclear power plants are outlined. The objectives of this work are to improve safety, provide a clearer understanding of safety significance, and ease NRC and industry administrative burdens. Line item improvements, bases, and implementation of the specifications are discussed.

  13. Place-Specific Computing

    DEFF Research Database (Denmark)

    Messeter, Jörn

    2009-01-01

    An increased interest in the notion of place has evolved in interaction design based on the proliferation of wireless infrastructures, developments in digital media, and a ‘spatial turn’ in computing. In this article, place-specific computing is suggested as a genre of interaction design...... that addresses the shaping of interactions among people, place-specific resources and global socio-technical networks, mediated by digital technology, and influenced by the structuring conditions of place. The theoretical grounding for place-specific computing is located in the meeting between conceptions...... examples of place-specific computing are presented from a series of pilot studies, conducted in close collaboration with design students in Malmö, Berlin, Cape Town and Rome, that generated 36 design concepts in the genre. Reflecting on these examples, issues in the design of place-specific computing...

  14. SPECIFIC DEGRADATION OF WATERSHEDS

    Institute of Scientific and Technical Information of China (English)

    Boubacar KANE; Pierre Y.JULIEN

    2007-01-01

    An extensive database of reservoir sedimentation surveys throughout continental United States is compiled and analyzed to determine specific degradation SD relationships as function of mean annual rainfall R, drainage area A, and watershed slope S. The database contains 1463 field measurements and specific degradation relationships are defined as function of A, R and S. Weak trends and significant variability in the data are noticeable. Specific degradation measurements are log normally distributed with respect to R, A, and S and 95% confidence intervals are determined accordingly. The accuracy of the predictions does not significantly increase as more independent variables are added to the regression analyses.

  15. FOXP3-specific immunity

    DEFF Research Database (Denmark)

    Andersen, Mads Hald

    2013-01-01

    Forkhead box P3 (FOXP3)-specific cytotoxic CD8(+) T cells are present among human peripheral blood mononuclear cells (PBMCs), especially in cancer patients. Such T lymphocytes are able not only to specifically recognize dendritic cells (DCs) that have been exposed to recombinant FOXP3 and regulat...... and regulatory T cells, but also to kill FOXP3(+) malignant T cells. The natural occurrence of FOXP3-specific cytotoxic T lymphocytes among human PBMCs suggests a general role for these cells in the complex network of immune regulation....

  16. The Learning Design Specification

    NARCIS (Netherlands)

    Olivier, Bill; Tattersall, Colin

    2005-01-01

    Olivier, B. & Tattersall, C. (2005). The Learning Design Specification In: Koper, R. & Tattersall, C., Learning Design: A Handbook on Modelling and Delivering Networked Education and Training (pp. 21-40). Berlin-Heidelberg: Springer Verlag.

  17. Place-Specific Computing

    DEFF Research Database (Denmark)

    Messeter, Jörn; Johansson, Michael

    An increased interest in the notion of place has evolved in interaction design. Proliferation of wireless infrastructure, developments in digital media, and a ‘spatial turn’ in computing provides the base for place-specific computing as a suggested new genre of interaction design. In the REcult...... project place- specific computing is explored through design oriented research. This article reports six pilot studies where design students have designed concepts for place-specific computing in Berlin (Germany), Cape Town (South Africa), Rome (Italy) and Malmö (Sweden). Background and arguments...... for place-specific computing as a genre of interaction design are described. A total number of 36 design concepts designed for 16 designated zones in the four cities are presented. An analysis of the design concepts is presented indicating potentials, possibilities and problems as directions for future...

  18. The Learning Design Specification

    NARCIS (Netherlands)

    Olivier, Bill; Tattersall, Colin

    2005-01-01

    Olivier, B. & Tattersall, C. (2005). The Learning Design Specification In: Koper, R. & Tattersall, C., Learning Design: A Handbook on Modelling and Delivering Networked Education and Training (pp. 21-40). Berlin-Heidelberg: Springer Verlag.

  19. Ontology Requirements Specification

    OpenAIRE

    Suárez-Figueroa, Mari Carmen; Gómez-Pérez, A.

    2012-01-01

    The goal of the ontology requirements specification activity is to state why the ontology is being built, what its intended uses are, who the end users are, and which requirements the ontology should fulfill. This chapter presents detailed methodological guidelines for specifying ontology requirements efficiently. These guidelines will help ontology engineers to capture ontology requirements and produce the ontology requirements specification document (ORSD). The ORSD will play a key role dur...

  20. Silencing effects of telomerase Cajal body protein 1-shRNA on human oral squamous cells%shRNA慢病毒对人口腔鳞状细胞癌细胞端粒酶卡侯体蛋白1基因的沉默效应

    Institute of Scientific and Technical Information of China (English)

    胡玲; 孙崇奎; 苟雅萍; 李燕; 肖丽英

    2013-01-01

    目的:探讨shRNA慢病毒对人口腔鳞状细胞癌Cal27细胞系端粒酶新蛋白,即端粒酶卡侯体蛋白1(TCAB1)基因的沉默效应,以期探索肿瘤基因治疗的新途径。方法:根据TCAB1基因,构建慢病毒shTCAB1-A~D,测定病毒滴度;用重组慢病毒体外感染Cal27细胞,荧光显微镜观察绿色荧光蛋白表达推断感染效率;半定量逆转录聚合酶链反应(RT-PCR)检测TCAB1 mRNA水平;Western blot检测TCAB1蛋白质的表达水平;四甲基偶氮唑盐比色(MTT)法检测细胞体外增殖能力;Transwell小室法检测细胞体外侵袭能力改变。结果:经PCR与测序鉴定证实成功构建靶向TCAB1的慢病毒RNAi载体,病毒滴度为1.5×108~3×108 U·mL-1。荧光显微镜观察显示,绝大部分细胞表达绿色荧光。与空白细胞对照组相比,4种不同的shTCAB1-A~D慢病毒感染组TCAB1 mRNA表达下调48.7%~62.0%;蛋白抑制率达45.6%~77.5%,shTCAB1-C组最明显。shTCAB1-C慢病毒能明显抑制Cal27细胞增殖能力,也能降低其侵袭能力,穿过人工基底膜的平均细胞数明显低于空白对照组及非特异性对照组,侵袭抑制率高达67.5%(P<0.05)。结论:成功构建并包装了端粒酶新蛋白TCAB1靶向的RNAi慢病毒,该病毒可高效感染人口腔鳞状细胞癌细胞,产生特异性的基因沉默效应。%Objective To investigate the inhibitory effects of telomerase Cajal body protein 1(TCAB1)-shRNA on oral squamous cell carcinoma Cal27 and to explore a novel cancer gene therapy approach by depletion of TCAB1 using RNA interference. Methods Four recombinant lentiviruses, called shTCAB1-A-D, which contain oligonucleotide(67 bp) targeting TCAB1 and green fluorescent protein EGFP Puro, were constructed based on different sites in the TCAB1 gene. A series of experiments were then carried out to demonstrate the silencing effects of the shTCAB1 lentiviruses on Cal27 cells. Semiquantitative

  1. Utility of the dual-specificity protein kinase TTK as a therapeutic target for intrahepatic spread of liver cancer.

    Science.gov (United States)

    Miao, Ruoyu; Wu, Yan; Zhang, Haohai; Zhou, Huandi; Sun, Xiaofeng; Csizmadia, Eva; He, Lian; Zhao, Yi; Jiang, Chengyu; Miksad, Rebecca A; Ghaziani, Tahereh; Robson, Simon C; Zhao, Haitao

    2016-09-13

    Therapies for primary liver cancer, the third leading cause of cancer-related death worldwide, remain limited. Following multi-omics analysis (including whole genome and transcriptome sequencing), we were able to identify the dual-specific protein kinase TTK as a putative new prognostic biomarker for liver cancer. Herein, we show that levels of TTK protein are significantly elevated in neoplastic tissues from a cohort of liver cancer patients, when compared with adjacent hepatic tissues. We also tested the utility of TTK targeted inhibition and have demonstrated therapeutic potential in an experimental model of liver cancer in vivo. Following lentiviral shRNA knockdown in several human liver cancer cell lines, we demonstrated that TTK boosts cell growth and promotes cell spreading; as well as protects against senescence and decreases autophagy. In an experimental animal model, we show that in vitro knockdown of TTK effectively blocks intrahepatic growth of human HCC xenografts. Furthermore, we note that, in vivo silencing of TTK, by systemically delivering TTK siRNAs to already tumor-bearing liver, limits intrahepatic spread of liver cancer cells. This intervention is associated with decreased tumor aggressiveness, as well as increased senescence and autophagy. Taken together, our data suggest that targeted TTK inhibition might have clinical utility as an adjunct therapy in management of liver cancer.

  2. Domain-Specific Multimodeling

    DEFF Research Database (Denmark)

    Hessellund, Anders

    Enterprise systems are complex artifacts. They are hard to build, manage, understand, and evolve. Existing software development paradigms fail to properly address challenges such as system size, domain complexity, and software evolution when development is scaled to enterprise systems. We propose...... domain-specific multimodeling as a development paradigm to tackle these challenges in a language-oriented manner. The different concerns of a system are conceptually separated and made explicit as independent domain-specific languages. This approach increases productivity and quality by raising...... the overall level of abstraction. It does, however, also introduce a new problem of coordinating multiple different languages in a single system. We call this problem the coordination problem. In this thesis, we present the coordination method for domain-specific multimodeling that explicitly targets...

  3. Comparative Test Case Specification

    DEFF Research Database (Denmark)

    Kalyanova, Olena; Heiselberg, Per

     This document includes a definition of the comparative test cases DSF200_3 and DSF200_4, which previously described in the comparative test case specification for the test cases DSF100_3 and DSF200_3 [Ref.1]....... This document includes a definition of the comparative test cases DSF200_3 and DSF200_4, which previously described in the comparative test case specification for the test cases DSF100_3 and DSF200_3 [Ref.1]....

  4. PLC-γ1 signaling plays a subtype-specific role in postbinding cell entry of influenza A virus.

    Science.gov (United States)

    Zhu, Liqian; Ly, Hinh; Liang, Yuying

    2014-01-01

    Host signaling pathways and cellular proteins play important roles in the influenza viral life cycle and can serve as antiviral targets. In this study, we report the engagement of host phosphoinositide-specific phospholipase γ1 (PLC-γ1) in mediating cell entry of influenza virus H1N1 but not H3N2 subtype. Both PLC-γ1-specific inhibitor and short hairpin RNA (shRNA) strongly suppress the replication of H1N1 but not H3N2 viruses in cell culture, suggesting that PLC-γ1 plays an important subtype-specific role in the influenza viral life cycle. Further analyses demonstrate that PLC-γ1 activation is required for viral postbinding cell entry. In addition, H1N1, but not H3N2, infection leads to the phosphorylation of PLC-γ1 at Ser 1248 immediately after infection and independent of viral replication. We have further shown that H1N1-induced PLC-γ1 activation is downstream of epidermal growth factor receptor (EGFR) signaling. Interestingly, both H1N1 and H3N2 infections activate EGFR, but only H1N1 infection leads to PLC-γ1 activation. Taking our findings together, we have identified for the first time the subtype-specific interplay of host PLC-γ1 signaling and H1N1 virus that is critical for viral uptake early in the infection. Our study provides novel insights into how virus interacts with the cellular signaling network by demonstrating that viral determinants can regulate how the host signaling pathways function in virally infected cells.

  5. Validation of glypican-3-specific scFv isolated from paired display/secretory yeast display library

    Directory of Open Access Journals (Sweden)

    Li Yonghai

    2012-05-01

    Full Text Available Abstract Background Glypican-3 (GPC3 is a heparan-sulfate proteoglycan frequently expressed on the cell membrane of malignant hepatocytes in hepatocellular carcinoma. The capacity for screening potential antibodies in vitro using human hepatocellular lines is critical to ensure binding to this highly post-translationally modified glycophosphatidylinositiol-linked protein. We hypothesized that we could utilize a recently described paired display/secretory yeast library to isolate human-derived scFv against glypican-3 for potential diagnostic and/or therapeutic application. Results Using two different biotinylated antigen targets, a synthesized 29mer fragment GPC3550-558 and a truncated GPC3368-548 fused with glutathione S-transferase (GST we enriched the yeast display library to greater than 30% target-specific yeast with both positive selection and depletion of streptavidin- and GST-specific clones. After cloning of scFv cDNA from the enriched sub-library, scFv specificity was validated by ELISA for binding to recombinant protein from prokaryotic and eukaryotic sources and ultimately naturally presented human protein on the cell membrane of human hepatocellular cell lines. Specificity was confirmed using non-expressing cell lines and shRNA knockdown. Ultimately, five unique scFv with affinity EC50 ranging from 5.0-110.9nM were identified. Conclusions Using a paired display/secretory yeast library, five novel and unique scFvs for potential humoral or chimeric therapeutic development in human hepatocellular carcinoma were isolated and characterized.

  6. Site-Specific Innovation

    DEFF Research Database (Denmark)

    Reeh, Henrik; Hemmersam, Peter

    2015-01-01

    , but also places for emerging lifestyles and cultural processes. In this transformation process, harbours provide arenas for a new urban dynamics, involving multiple sectors and functions in society, as opposed to the monoculturalism prevailing in the ages of the industrial city. To us, the critical...... question is how innovation may contribute to urban life and site-specific qualities....

  7. Acquiring specific interpreting competence

    Directory of Open Access Journals (Sweden)

    Jana Zidar Forte

    2012-12-01

    Full Text Available In postgraduate interpreter training, the main objective of the course is to help trainees develop various competences, from linguistic, textual and cultural competence, to professional and specific interpreting competence. For simultaneous interpreting (SI, the main focus is on mastering the SI technique and strategies as well as on developing and strengthening communicative skills, which is discussed and illustrated with examples in the present paper. First, a brief overview is given of all the necessary competences of a professional interpreter with greater emphasis on specific interpreting competence for SI. In the second part of the paper, various approaches are described in terms of acquiring specific skills and strategies, specifically through a range of exercises. Besides interpreting entire speeches, practical courses should also consist of targeted exercises, which help trainees develop suitable coping strategies and mechanisms (later on almost automatisms, while at the same time "force" them to reflect on their individual learning process and interpreting performance. This provides a solid base on which trained interpreters can progress and develop their skills also after joining the professional sphere.

  8. Site-Specific Innovation

    DEFF Research Database (Denmark)

    Reeh, Henrik; Hemmersam, Peter

    2015-01-01

    Currently, cities across the Northern European region are actively redeveloping their former industrial harbours. Indeed, harbours areas are essential in the long-term transition from industrial to information and experience societies; harbours are becoming sites for new businesses and residences...... question is how innovation may contribute to urban life and site-specific qualities....

  9. IDC System Specification Document.

    Energy Technology Data Exchange (ETDEWEB)

    Clifford, David J.

    2014-12-01

    This document contains the system specifications derived to satisfy the system requirements found in the IDC System Requirements Document for the IDC Reengineering Phase 2 project. Revisions Version Date Author/Team Revision Description Authorized by V1.0 12/2014 IDC Reengineering Project Team Initial delivery M. Harris

  10. LOCKE Detailed Specification Tables

    CERN Document Server

    Menezo, Lucia G; Gregorio, Jose-Angel

    2012-01-01

    This document shows the detailed specification of LOCKE coherence protocol for each cache controller, using a table-based technique. This representation provides clear, concise visual information yet includes sufficient detail (e.g., transient states) arguably lacking in the traditional, graphical form of state diagrams.

  11. Children's specific fears.

    Science.gov (United States)

    Meltzer, H; Vostanis, P; Dogra, N; Doos, L; Ford, T; Goodman, R

    2009-11-01

    Most children experience some degree of fear during their development. Specific fears are considered as an appropriate response provided that they are proportionate to the intensity of the perceived threat. Our aim is to present the prevalence of specific fears among children in the Great Britain, their socio-demographic correlates, in particular their association with ethnicity. Data on the child's experience of specific fears were obtained from parents of a national representative sample of 5- to 16-year-olds using the Development and Well-Being Assessment. Biographic, socio-demographic and socioeconomic characteristics of the child and the family were included in the questionnaire. About one-third of children were assessed by their parents as having at least one of 12 specific fears. The most commonly reported fears were animals (11.6%), blood/injections (10.8%) and the dark (6.3%). Just less than 1% of all children were assessed according to International Classification of Diseases research diagnostic criteria as having a specific phobia. Biographic, socio-demographic and socioeconomic factors were independently associated with a greater likelihood of a child having particular fears. The most marked associations were fears of the dark, loud noises, imagined supernatural beings in younger children and fear of animals among girls and all non-white groups. Although fears are only labelled as phobias when they impair functioning and interfere with life, they can cause personal distress to children and also can interfere with their daily activities. Children's fears differ in nature across different ethnic groups. Culturally mediated beliefs, values and traditions may play a role in their expression.

  12. Specificity in LSP

    Directory of Open Access Journals (Sweden)

    Thomas N. Huckin

    2003-04-01

    Full Text Available The defining feature of our field is the teaching and use of language for specific purposes. Not surprisingly, this has led to a debate over just how specific those purposes should be. The debate is longstanding, stirred initially by Hutchinson and Waters' classic 1980 article, 'ESP at the Crossroads' and reinvigorated most recently by Hyland (2002, who argues that the field has drifted away from specificity, becoming too generalized and diffuse. This paper lays out the arguments on both sides and then stakes out a position that draws from each of them. While agreeing with some of Hyland's criticisms of the 'wide-angle' position (for example, that generalized LSP can fail to appreciate the distinct linguistic and rhetorical features of specialized discourses, the paper criticizes his 'narrow-angle' position as well by pointing out that it can easily lead to a teacher-centered prescriptivism and to an overly rigid focus on certain forms and tasks at the expense of others. Furthermore, such an approach fails to prepare students for the unpredictable new forms of communication that await them in their professional careers. In general, a teachercentered approach, no matter how specific, is unlikely to have the pedagogical effectiveness of a student-centered approach, especially in heterogeneous classes. Specificity, it is argued, must ultimately be supplied by the student, not by the teacher, for it is the student more than the LSP teacher who is in the process of becoming an insider and whose interests are best served by becoming an astute analyst of the specialist discourse. The teacher's role should be that of facilitator, instructing students in analytic strategies, both rhetorical and textual. The paper concludes with a number of illustrations in how this can be done, drawing especially on work in genre study.

  13. Allergen Specific Immunotherapy

    Directory of Open Access Journals (Sweden)

    Şükrü Çekiç

    2015-04-01

    Full Text Available Allergen specific immunotherapy (SIT is the only treatment that can provide a cure for allergic disorders. This treatment is based on development of immune tolerance by exposure to allergen in repetitive and increasing doses. It is tertiary to avoidance of allergen and pharmacotherapy. Allergens used for immunotherapy, must be confirmed by skin prick test or specific IgE and must be applied in supervision of allergy specialists. Studies show that immunotherapy, improve asthma symptoms, decreases drug consumption, prevent development of asthma in rhinitis patients and reduce new sensitizations. Common side effects diminished with the usage of standardized allergen solutions. It is contraindicated in severe asthma. Though it is recommended to avoid immunotherapy in patients using beta blockers and ACE inhibitors, immunotherapy can be considered in mandatory situations regarding possible benefits and harms. Most common ways of administration are subcutaneous and sublingual; new methods such as epicutaneous and intralymphatic injections are currently being studied.

  14. Safety detaching hook specification.

    CSIR Research Space (South Africa)

    Roux, JD

    1999-05-01

    Full Text Available from the mean surface' and measured in micrometres (µm). Regulations Regulations pertaining to an act or amended act as promulgated by the Government of South Africa. SAATCA Southern African Auditor and Training Certification Association. SABS... of this specification have been adhered to. Technical Assessment An audit report on a manufacturer's systems for the management of product quality, issued by an independent auditor registered with SAATCA. 6 2. REQUIREMENTS 2.1 Design Requirements a) General...

  15. Empirical Test Case Specification

    DEFF Research Database (Denmark)

    Kalyanova, Olena; Heiselberg, Per

    This document includes the empirical specification on the IEA task of evaluation building energy simulation computer programs for the Double Skin Facades (DSF) constructions. There are two approaches involved into this procedure, one is the comparative approach and another is the empirical one. I....... In the comparative approach the outcomes of different software tools are compared, while in the empirical approach the modelling results are compared with the results of experimental test cases....

  16. Comparative Test Case Specification

    DEFF Research Database (Denmark)

    Kalyanova, Olena; Heiselberg, Per

    This document includes the specification on the IEA task of evaluation building energy simulation computer programs for the Double Skin Facades (DSF) constructions. There are two approaches involved into this procedure, one is the comparative approach and another is the empirical one. In the comp....... In the comparative approach the outcomes of different software tools are compared, while in the empirical approach the modelling results are compared with the results of experimental test cases. The comparative test cases include: ventilation, shading and geometry....

  17. Allergen-specific immunotherapy

    Directory of Open Access Journals (Sweden)

    Moote William

    2011-11-01

    Full Text Available Abstract Allergen-specific immunotherapy is a potentially disease-modifying therapy that is effective for the treatment of allergic rhinitis/conjunctivitis, allergic asthma and stinging insect hypersensitivity. However, despite its proven efficacy in these conditions, it is frequently underutilized in Canada. The decision to proceed with allergen-specific immunotherapy should be made on a case-by-case basis, taking into account individual patient factors such as the degree to which symptoms can be reduced by avoidance measures and pharmacological therapy, the amount and type of medication required to control symptoms, the adverse effects of pharmacological treatment, and patient preferences. Since this form of therapy carries the risk of anaphylactic reactions, it should only be prescribed by physicians who are adequately trained in the treatment of allergy. Furthermore, injections must be given under medical supervision in clinics that are equipped to manage anaphylaxis. In this article, the authors review the indications and contraindications, patient selection criteria, and the administration, safety and efficacy of allergen-specific immunotherapy.

  18. Specification of Concurrent Objects

    DEFF Research Database (Denmark)

    Sørensen, Morten U.

    relation over two objects and an event. In the model, objects can be composed by parallel composition, encapsulation, and hiding of operations. Refinement between objects is defined as fair trace inclusion.A specification language is presented where objects can be specified operationally by abstract......Concurrent objects are named concurrent processes that interact by invoking each other's operations. We describe how such concurrent objects can be specified, how objects can be composed, and how it can be shown that one object refines another.First a model is defined, based on a transition...

  19. Disease specific protein corona

    Science.gov (United States)

    Rahman, M.; Mahmoudi, M.

    2015-03-01

    It is now well accepted that upon their entrance into the biological environments, the surface of nanomaterials would be covered by various biomacromolecules (e.g., proteins and lipids). The absorption of these biomolecules, so called `protein corona', onto the surface of (nano)biomaterials confers them a new `biological identity'. Although the formation of protein coronas on the surface of nanoparticles has been widely investigated, there are few reports on the effect of various diseases on the biological identity of nanoparticles. As the type of diseases may tremendously changes the composition of the protein source (e.g., human plasma/serum), one can expect that amount and composition of associated proteins in the corona composition may be varied, in disease type manner. Here, we show that corona coated silica and polystyrene nanoparticles (after interaction with in the plasma of the healthy individuals) could induce unfolding of fibrinogen, which promotes release of the inflammatory cytokines. However, no considerable releases of inflammatory cytokines were observed for corona coated graphene sheets. In contrast, the obtained corona coated silica and polystyrene nanoparticles from the hypofibrinogenemia patients could not induce inflammatory cytokine release where graphene sheets do. Therefore, one can expect that disease-specific protein coronas can provide a novel approach for applying nanomedicine to personalized medicine, improving diagnosis and treatment of different diseases tailored to the specific conditions and circumstances.

  20. ADANS database specification

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-01-16

    The purpose of the Air Mobility Command (AMC) Deployment Analysis System (ADANS) Database Specification (DS) is to describe the database organization and storage allocation and to provide the detailed data model of the physical design and information necessary for the construction of the parts of the database (e.g., tables, indexes, rules, defaults). The DS includes entity relationship diagrams, table and field definitions, reports on other database objects, and a description of the ADANS data dictionary. ADANS is the automated system used by Headquarters AMC and the Tanker Airlift Control Center (TACC) for airlift planning and scheduling of peacetime and contingency operations as well as for deliberate planning. ADANS also supports planning and scheduling of Air Refueling Events by the TACC and the unit-level tanker schedulers. ADANS receives input in the form of movement requirements and air refueling requests. It provides a suite of tools for planners to manipulate these requirements/requests against mobility assets and to develop, analyze, and distribute schedules. Analysis tools are provided for assessing the products of the scheduling subsystems, and editing capabilities support the refinement of schedules. A reporting capability provides formatted screen, print, and/or file outputs of various standard reports. An interface subsystem handles message traffic to and from external systems. The database is an integral part of the functionality summarized above.

  1. Tissue specific electrochemical fingerprinting.

    Directory of Open Access Journals (Sweden)

    Pavlina Sobrova

    Full Text Available BACKGROUND: Proteomics and metalloproteomics are rapidly developing interdisciplinary fields providing enormous amounts of data to be classified, evaluated and interpreted. Approaches offered by bioinformatics and also by biostatistical data analysis and treatment are therefore of extreme interest. Numerous methods are now available as commercial or open source tools for data processing and modelling ready to support the analysis of various datasets. The analysis of scientific data remains a big challenge, because each new task sets its specific requirements and constraints that call for the design of a targeted data pre-processing approach. METHODOLOGY/PRINCIPAL FINDINGS: This study proposes a mathematical approach for evaluating and classifying datasets obtained by electrochemical analysis of metallothionein in rat 9 tissues (brain, heart, kidney, eye, spleen, gonad, blood, liver and femoral muscle. Tissue extracts were heated and then analysed using the differential pulse voltammetry Brdicka reaction. The voltammograms were subsequently processed. Classification models were designed making separate use of two groups of attributes, namely attributes describing local extremes, and derived attributes resulting from the level=5 wavelet transform. CONCLUSIONS/SIGNIFICANCE: On the basis of our results, we were able to construct a decision tree that makes it possible to distinguish among electrochemical analysis data resulting from measurements of all the considered tissues. In other words, we found a way to classify an unknown rat tissue based on electrochemical analysis of the metallothionein in this tissue.

  2. Site-specific

    Directory of Open Access Journals (Sweden)

    Adel M.E. Mohamed

    2013-06-01

    Full Text Available In order to quantify the near-surface seismic properties (P- and S-wave velocities, and the dynamic elastic properties with respect to the depth at a specific area (6th of October club, we conducted a non-invasive and low cost active seismic survey. The primary wave velocity is determined by conducting a P-wave shallow seismic refraction survey. The dispersive characteristics of Rayleigh type surface waves were utilized for imaging the shallow subsurface layers by estimating the 1D (depth and 2D (depth and surface location shear wave velocities. The reliability of the Multi-channel Analysis of Surface Waves (MASW depends on the accurate determination of phase velocities for horizontally traveling fundamental mode Rayleigh waves. Consequently, the elastic properties are evaluated empirically. The Vs30 (average shear wave velocity down to 30 m depth, which is obtained from the MASW technique, plays a critical role in evaluating the site response of the upper 30 m depth. The distribution of the obtained values of Vs30 through the studied area demonstrates site classes of C and D, according to the NEHRP (National Earthquake Hazard Reduction Program and IBC (International Building Code standards.

  3. Inhibition of N1-Src kinase by a specific SH3 peptide ligand reveals a role for N1-Src in neurite elongation by L1-CAM

    Science.gov (United States)

    Keenan, Sarah; Wetherill, Sarah J.; Ugbode, Christopher I.; Chawla, Sangeeta; Brackenbury, William J.; Evans, Gareth J. O.

    2017-01-01

    In the mammalian brain the ubiquitous tyrosine kinase, C-Src, undergoes splicing to insert short sequences in the SH3 domain to yield N1- and N2-Src. We and others have previously shown that the N-Srcs have altered substrate specificity and kinase activity compared to C-Src. However, the exact functions of the N-Srcs are unknown and it is likely that N-Src signalling events have been misattributed to C-Src because they cannot be distinguished by conventional Src inhibitors that target the kinase domain. By screening a peptide phage display library, we discovered a novel ligand (PDN1) that targets the unique SH3 domain of N1-Src and inhibits N1-Src in cells. In cultured neurons, PDN1 fused to a fluorescent protein inhibited neurite outgrowth, an effect that was mimicked by shRNA targeting the N1-Src microexon. PDN1 also inhibited L1-CAM-dependent neurite elongation in cerebellar granule neurons, a pathway previously shown to be disrupted in Src−/− mice. PDN1 therefore represents a novel tool for distinguishing the functions of N1-Src and C-Src in neurons and is a starting point for the development of a small molecule inhibitor of N1-Src. PMID:28220894

  4. Assessing somatic hypermutation in Ramos B cells after overexpression or knockdown of specific genes.

    Science.gov (United States)

    Upton, Dana C; Unniraman, Shyam

    2011-11-01

    -activated cell scanning (FACS) provides a quick read-out for the level of SHM. A more quantitative measurement of SHM can be obtained by directly sequencing the antibody genes. Since Ramos cells are difficult to transfect, we produce stable derivatives that have increased or lowered expression of an individual gene by infecting cells with retroviral or lentiviral constructs that contain either an overexpression cassette or a short hairpin RNA (shRNA), respectively. Here, we describe how we infect Ramos cells and then use these cells to investigate the role of specific genes on SHM (Figure 1).

  5. Fat-specific Protein 27/CIDEC Promotes Development of Alcoholic Steatohepatitis in Mice and Humans

    Science.gov (United States)

    Xu, Ming-Jiang; Cai, Yan; Wang, Hua; Altamirano, José; Chang, Binxia; Bertola, Adeline; Odena, Gemma; Lu, Jim; Tanaka, Naoki; Matsusue, Kimihiko; Matsubara, Tsutomu; Mukhopadhyay, Partha; Kimura, Shioko; Pacher, Pal; Gonzalez, Frank J; Bataller, Ramon; Gao, Bin

    2015-01-01

    Background & Aims Alcoholic steatohepatitis (ASH) is the progressive form of alcoholic liver disease and may lead to cirrhosis and hepatocellular carcinoma. We studied mouse models and human tissues to identify molecules associated with ASH progression, and focused on mouse fat-specific protein 27 (FSP-27)/human cell death-inducing DFF45-like effector C (CIDEC) protein, which is expressed in white adipose tissues and promotes formation of fat droplets. Methods C57BL/6N mice or mice with hepatocyte-specific disruption of Fsp27 (Fsp27Hep−/− mice) were fed the Lieber-Decarli ethanol liquid diet (5% ethanol) for 10 days to 12 weeks, followed by 1 or multiple binges of ethanol (5 or 6 g/kg) during the chronic feeding. Some mice were given an inhibitor of the peroxisome proliferator-activated receptor-γ (PPARG) (GW9662). Adenoviral vectors were used to express transgenes or small hairpin (sh) RNAs in cultured hepatocytes and in mice. Liver tissue samples were collected from ethanol-fed mice or from 31 patients with alcoholic hepatitis (AH) with biopsy-proved ASH and analyzed by histologic, immunohistochemical, transcriptome, immunoblot, and real-time PCR analyses. Results Chronic-plus-binge ethanol feeding of mice, which mimics the drinking pattern of patients with AH, produced severe ASH and mild fibrosis. Microarray analyses revealed similar alterations in expression of many hepatic genes in ethanol-fed mice and humans with ASH, including upregulation of mouse Fsp27 (also called Cidec) and human CIDEC. Fsp27Hep−/− mice and mice given injections of adenovirus-Fsp27shRNA had markedly reduced ASH following chronic-plus-binge ethanol feeding. Inhibition of PPARG and cyclic AMP-responsive element binding protein H (CREBH) prevented the increases in Fsp27α and FSP27β mRNAs, respectively, and reduced liver injury in this chronic-plus-binge ethanol feeding model. Overexpression of FSP27 and ethanol exposure had synergistic effects in inducing production of

  6. Teaching English for Specific Purposes

    Directory of Open Access Journals (Sweden)

    Nijolė Netikšienė

    2011-04-01

    Full Text Available Teaching English for Specific Purposes and General English is analysed in the article. The scientific approach of a scientist M. Rosenberg is presented. The experience of teaching English for Specific Purposesat VGTU is alsopresented. The ideas and teaching methods from the classes of general English can be transferred to the classes of English for Specific Purposes.

  7. Characterization of a novel small molecule subtype specific estrogen-related receptor alpha antagonist in MCF-7 breast cancer cells.

    Directory of Open Access Journals (Sweden)

    Michael J Chisamore

    Full Text Available BACKGROUND: The orphan nuclear receptor estrogen-related receptor alpha (ERRalpha is a member of the nuclear receptor superfamily. It was identified through a search for genes encoding proteins related to estrogen receptor alpha (ERalpha. An endogenous ligand has not been found. Novel ERRalpha antagonists that are highly specific for binding to the ligand binding domain (LBD of ERRalpha have been recently reported. Research suggests that ERRalpha may be a novel drug target to treat breast cancer and/or metabolic disorders and this has led to an effort to characterize the mechanisms of action of N-[(2Z-3-(4,5-dihydro-1,3-thiazol-2-yl-1,3-thiazolidin-2-yl idene]-5H dibenzo[a,d][7]annulen-5-amine, a novel ERRalpha specific antagonist. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate this ERRalpha ligand inhibits ERRalpha transcriptional activity in MCF-7 cells by luciferase assay but does not affect mRNA levels measured by real-time RT-PCR. Also, ERalpha (ESR1 mRNA levels were not affected upon treatment with the ERRalpha antagonist, but other ERRalpha (ESRRA target genes such as pS2 (TFF1, osteopontin (SPP1, and aromatase (CYP19A1 mRNA levels decreased. In vitro, the ERRalpha antagonist prevents the constitutive interaction between ERRalpha and nuclear receptor coactivators. Furthermore, we use Western blots to demonstrate ERRalpha protein degradation via the ubiquitin proteasome pathway is increased by the ERRalpha-subtype specific antagonist. We demonstrate by chromatin immunoprecipitation (ChIP that the interaction between ACADM, ESRRA, and TFF1 endogenous gene promoters and ERRalpha protein is decreased when cells are treated with the ligand. Knocking-down ERRalpha (shRNA led to similar genomic effects seen when MCF-7 cells were treated with our ERRalpha antagonist. CONCLUSIONS/SIGNIFICANCE: We report the mechanism of action of a novel ERRalpha specific antagonist that inhibits transcriptional activity of ERRalpha, disrupts the constitutive

  8. Specification-Based Testing Via Domain Specific Language

    Directory of Open Access Journals (Sweden)

    Sroka Michal

    2014-12-01

    Full Text Available The article presents tCF (testCaseFramework - a domain specific language with corresponding toolchain for specification-based software testing of embedded software. tCF is designed for efficient preparation of maintainable and intelligible test cases and for testing process automation, as it allows to generate platform specific test cases for various testing levels. The article describes the essential parts of the tCF meta-model and the applied concept of platform specific test cases generators.

  9. Triptolide inhibits transcription of hTERT through down-regulation of transcription factor specificity protein 1 in primary effusion lymphoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Long, Cong; Wang, Jingchao [Department of Pathogen Biology, School of Basic Medical Sciences, Wuhan University, Wuhan, 430071 (China); Guo, Wei [Department of Pathology and Physiology, School of Basic Medical Sciences, Wuhan University, Wuhan, 430071 (China); Wang, Huan; Wang, Chao; Liu, Yu [Department of Pathogen Biology, School of Basic Medical Sciences, Wuhan University, Wuhan, 430071 (China); Sun, Xiaoping, E-mail: xsun6@whu.edu.cn [Department of Pathogen Biology, School of Basic Medical Sciences, Wuhan University, Wuhan, 430071 (China); State Key Laboratory of Virology, Wuhan University, Wuhan, 430072 (China)

    2016-01-01

    Primary effusion lymphoma (PEL) is a rare and aggressive non-Hodgkin's lymphoma. Human telomerase reverse transcriptase (hTERT), a key component responsible for the regulation of telomerase activity, plays important roles in cellular immortalization and cancer development. Triptolide purified from Tripterygium extracts displays a broad-spectrum bioactivity profile, including immunosuppressive, anti-inflammatory, and anti-tumor. In this study, it is investigated whether triptolide reduces hTERT expression and suppresses its activity in PEL cells. The mRNA and protein levels of hTERT were examined by real time-PCR and Western blotting, respectively. The activity of hTERT promoter was determined by Dual luciferase reporter assay. Our results demonstrated that triptolide decreased expression of hTERT at both mRNA and protein levels. Further gene sequence analysis indicated that the activity of hTERT promoter was suppressed by triptolide. Triptolide also reduced the half-time of hTERT. Additionally, triptolide inhibited the expression of transcription factor specificity protein 1(Sp1) in PEL cells. Furthermore, knock-down of Sp1 by using specific shRNAs resulted in down-regulation of hTERT transcription and protein expression levels. Inhibition of Sp1 by specific shRNAs enhanced triptolide-induced cell growth inhibition and apoptosis. Collectively, our results demonstrate that the inhibitory effect of triptolide on hTERT transcription is possibly mediated by inhibition of transcription factor Sp1 in PEL cells. - Highlights: • Triptolide reduces expression of hTERT by decreasing its transcription level. • Triptolide reduces promoter activity and stability of hTERT. • Triptolide down-regulates expression of Sp1. • Special Sp1 shRNAs inhibit transcription and protein expression of hTERT. • Triptolide and Sp1 shRNA2 induce cell proliferation inhibition and apoptosis.

  10. Ion specificities of artificial macromolecules.

    Science.gov (United States)

    Liu, Lvdan; Kou, Ran; Liu, Guangming

    2016-12-21

    Artificial macromolecules are well-defined synthetic polymers, with a relatively simple structure as compared to naturally occurring macromolecules. This review focuses on the ion specificities of artifical macromolecules. Ion specificities are influenced by solvent-mediated indirect ion-macromolecule interactions and also by direct ion-macromolecule interactions. In aqueous solutions, the role of water-mediated indirect ion-macromolecule interactions will be discussed. The addition of organic solvents to aqueous solutions significantly changes the ion specificities due to the formation of water-organic solvent complexes. For direct ion-macromolecule interactions, we will discuss specific ion-pairing interactions for charged macromolecules and specific ion-neutral site interactions for uncharged macromolecules. When the medium conditions change from dilute solutions to crowded environments, the ion specificities can be modified by either the volume exclusion effect, the variation of dielectric constant, or the interactions between ions, macromolecules, and crowding agents.

  11. The Order Specification Decoupling Line

    DEFF Research Database (Denmark)

    Hansen, Benjamin Loer; Hvam, Lars

    2004-01-01

    find some welding specifications or product descriptions which are standard for every order, while there may be other specifications, such as bill-of-materials and drawings, which are customized. Through a use of this concept it becomes possible to discuss different levels of OSDL and different variant......This paper discusses the important issue in build-to-order production of separating innovative long term engineering processes from day-to-day variant specification processes, i.e. the processes creating specifications such as bill-of-materials, drawings, text descriptions, routings etc. The paper...... proposes and discusses the term Order Specification Decoupling Line (OSDL), which is to be used to explain the separation of industrial specifications into some information created prior to order acquisition and some information created during order acquisition and order fulfillment. For instance one may...

  12. Action-Specific Effects Underwater

    OpenAIRE

    Witt, Jessica; Schuck, Donald M; Taylor, J. Eric T.

    2011-01-01

    Action-specific effects on perception are apparent in terrestrial environments. For example, targets that require more effort to walk, jump, or throw to look farther away than when the targets require less effort. Here, we examined whether action-specific effects would generalize to an underwater environment. Instead, perception might be geometrically precise, rather than action-specific, in an environment that is novel from an evolutionary perspective. We manipulated ease to swim by giving p...

  13. [Multilingualism and specific language impairment].

    Science.gov (United States)

    Arkkila, Eva; Smolander, Sini; Laasonen, Marja

    2013-01-01

    Specific language impairment is one of the most common developmental disturbances in childhood. With the increase of the foreign language population group an increasing number of children assimilating several languages and causing concern in language development attend clinical examinations. Knowledge of factors underlying the specific language impairment and the specific impairment in general, special features of language development of those learning several languages, as well as the assessment and support of the linguistic skills of a multilingual child is essential. The risk of long-term problems and marginalization is high for children having specific language impairment.

  14. Hanford Site environmental management specification

    Energy Technology Data Exchange (ETDEWEB)

    Grygiel, M.L.

    1998-06-10

    The US Department of Energy, Richland Operations Office (RL) uses this Hanford Site Environmental Management Specification (Specification) to document top-level mission requirements and planning assumptions for the prime contractors involved in Hanford Site cleanup and infrastructure activities under the responsibility of the US Department of Energy, Office of Environmental Management. This Specification describes at a top level the activities, facilities, and infrastructure necessary to accomplish the cleanup of the Hanford Site and assigns this scope to Site contractors and their respective projects. This Specification also references the key National Environmental Policy Act of 1969 (NEPA), Comprehensive Environmental Response, Compensation, and Liability Act of 1980 (CERCLA), and safety documentation necessary to accurately describe the cleanup at a summary level. The information contained in this document reflects RL`s application of values, priorities, and critical success factors expressed by those involved with and affected by the Hanford Site project. The prime contractors and their projects develop complete baselines and work plans to implement this Specification. These lower-level documents and the data that support them, together with this Specification, represent the full set of requirements applicable to the contractors and their projects. Figure 1-1 shows the relationship of this Specification to the other basic Site documents. Similarly, the documents, orders, and laws referenced in this specification represent only the most salient sources of requirements. Current and contractual reference data contain a complete set of source documents.

  15. Response to "Transfer or Specificity?"

    Science.gov (United States)

    Miller, Judith

    2007-01-01

    This article presents the author's response to "Transfer or Specificity?" and reports a research that supports a strong case for a fundamental motor skill as a precursor to two sport specific skills as in Gallahue and Ozmun's (2002) theoretical model of motor development. Reported changes in performance of the overarm throw are…

  16. Performance Specifications for Occupational Programs.

    Science.gov (United States)

    Maryland State Dept. of Education, Baltimore. Div. of Career Technology and Adult Learning.

    This document lists and discusses the development of Maryland's performance specifications for occupational programs. The introduction explains the process used to develop performance standards and specifications for 10 career cluster majors that were identified by a task force of educators and employers as high-demand occupational areas in…

  17. Functional specifications for mathematical computations

    Energy Technology Data Exchange (ETDEWEB)

    Boyle, J.M. (Argonne National Lab., IL (USA)); Harmer, T.J. (Queen' s Univ., Belfast, Northern Ireland (UK). Dept. of Computer Science)

    1991-01-01

    Are functional programs useful for specifying numerical computations We believe they certainly are, despite the long-established tradition of using procedural languages for such computations. We have prepared a pure functional specification for an algorithm that solves one-dimensional hyperbolic partial differential equations (PDEs). Using automated program transformations, we have derived a Fortran program from this specification that executes faster on a CRAY X-MP than does the hand-written Fortran implementation of the same algorithm. We discuss the development of the initial specification for the one-dimensional problem and its evolution into a second specification for solving multidimensional hyperbolic PDEs. In this second specification, the dimensionality of the problem is completely parameterized and is given by specifying the set of neighbors of a cell in the grid. Thus, programs can be derived from this specification to solve hyperbolic PDEs of any given dimensionality. Our goal is to elucidate how we approach specifying numerical computations in the functional style and to show how we take advantage of the modularity and abstractness of functional programming to obtain a very high-level representation of the algorithm. We also briefly discuss transformational derivation of efficient programs from such specifications. 13 refs., 1 tab.

  18. Epithelial Expression of Human ABO Blood Group Genes Is Dependent upon a Downstream Regulatory Element Functioning through an Epithelial Cell-specific Transcription Factor, Elf5.

    Science.gov (United States)

    Sano, Rie; Nakajima, Tamiko; Takahashi, Yoichiro; Kubo, Rieko; Kobayashi, Momoko; Takahashi, Keiko; Takeshita, Haruo; Ogasawara, Kenichi; Kominato, Yoshihiko

    2016-10-21

    The human ABO blood group system is of great importance in blood transfusion and organ transplantation. The ABO system is composed of complex carbohydrate structures that are biosynthesized by A- and B-transferases encoded by the ABO gene. However, the mechanisms regulating ABO gene expression in epithelial cells remain obscure. On the basis of DNase I-hypersensitive sites in and around ABO in epithelial cells, we prepared reporter plasmid constructs including these sites. Subsequent luciferase assays and histone modifications indicated a novel positive regulatory element, designated the +22.6-kb site, downstream from ABO, and this was shown to enhance ABO promoter activity in an epithelial cell-specific manner. Expression of ABO and B-antigen was reduced in gastric cancer KATOIII cells by biallelic deletion of the +22.6-kb site using the CRISPR/Cas9 system. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay demonstrated that the site bound to an epithelial cell-specific transcription factor, Elf5. Mutation of the Ets binding motifs to abrogate binding of this factor reduced the regulatory activity of the +22.6-kb site. Furthermore, ELF5 knockdown with shRNA reduced both endogenous transcription from ABO and B-antigen expression in KATOIII cells. Thus, Elf5 appeared to be involved in the enhancer potential of the +22.6-kb site. These results support the contention that ABO expression is dependent upon a downstream positive regulatory element functioning through a tissue-restricted transcription factor, Elf5, in epithelial cells.

  19. Patient-Specific Computational Modeling

    CERN Document Server

    Peña, Estefanía

    2012-01-01

    This book addresses patient-specific modeling. It integrates computational modeling, experimental procedures, imagine clinical segmentation and mesh generation with the finite element method (FEM) to solve problems in computational biomedicine and bioengineering. Specific areas of interest include cardiovascular problems, ocular and muscular systems and soft tissue modeling. Patient-specific modeling has been the subject of serious research over the last seven years and interest in the area is continually growing and this area is expected to further develop in the near future.

  20. A role for Insulin-like growth factor 2 in specification of the fast skeletal muscle fibre

    Directory of Open Access Journals (Sweden)

    Ting Tao

    2007-06-01

    Full Text Available Abstract Background Fibre type specification is a poorly understood process beginning in embryogenesis in which skeletal muscle myotubes switch myosin-type to establish fast, slow and mixed fibre muscle groups with distinct function. Growth factors are required to establish slow fibres; it is unknown how fast twitch fibres are specified. Igf-2 is an embryonically expressed growth factor with established in vitro roles in skeletal muscle. Its localisation and role in embryonic muscle differentiation had not been established. Results Between E11.5 and E15.5 fast Myosin (FMyHC localises to secondary myotubes evenly distributed throughout the embryonic musculature and gradually increasing in number so that by E15.5 around half contain FMyHC. The Igf-2 pattern closely correlates with FMyHC from E13.5 and peaks at E15.5 when over 90% of FMyHC+ myotubes also contain Igf-2. Igf-2 lags FMyHC and it is absent from muscle myotubes until E13.5. Igf-2 strongly down-regulates by E17.5. A striking feature of the FMyHC pattern is its increased heterogeneity and attenuation in many fibres from E15.5 to day one after birth (P1. Transgenic mice (MIG which express Igf-2 in all of their myotubes, have increased FMyHC staining, a higher proportion of FMyHC+ myotubes and loose their FMyHC staining heterogeneity. In Igf-2 deficient mice (MatDi FMyHC+ myotubes are reduced to 60% of WT by E15.5. In vitro, MIG induces a 50% excess of FMyHC+ and a 30% reduction of SMHyC+ myotubes in C2 cells which can be reversed by Igf-2-targeted ShRNA resulting in 50% reduction of FMyHC. Total number of myotubes was not affected. Conclusion In WT embryos the appearance of Igf-2 in embryonic myotubes lags FMyHC, but by E15.5 around 45% of secondary myotubes contain both proteins. Forced expression of Igf-2 into all myotubes causes an excess, and absence of Igf-2 suppresses, the FMyHC+ myotube component in both embryonic muscle and differentiated myoblasts. Igf-2 is thus required, not for

  1. Youth job market specific features

    Directory of Open Access Journals (Sweden)

    Evgeniya Yu. Zhuravleva

    2011-05-01

    Full Text Available The article considers youth job market peculiarities, its specific features and regulation means, determines theoretical and application tasks of qualitative and quantitative comparison of vocations, which are highly in demand at the job market.

  2. GFF Specification for SAR Systems

    Energy Technology Data Exchange (ETDEWEB)

    Sweet, Nathan [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Bielek, Timothy P. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Matthews, John D. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Coakley, Henry John [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Pan, Len [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2014-08-01

    This document provides the basic GFF format definition for a GFF file. This document covers the contents of the main header and the basic GFF file structure. The contents of specific extensions are covered in a separate document.

  3. Eternity Variables to Simulate Specifications

    NARCIS (Netherlands)

    Hesselink, WH; Boiten, EA; Moller, B

    2002-01-01

    Simulation of specifications is introduced as a unification and generalization of refinement mappings, history variables, forward simulations, prophecy variables, and backward simulations. Eternity variables are introduced as a more powerful alternative for prophecy variables and backward simulation

  4. EEG in Specific Language Impairment

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2008-11-01

    Full Text Available The value of routine wake electroencephalography in children with specific language impairment was reviewed retrospectively in 111 children examined over a 10-year interval at Montreal Children’s Hospital, Quebec, Çanada.

  5. CCL22-specific T Cells

    DEFF Research Database (Denmark)

    Martinenaite, Evelina; Munir Ahmad, Shamaila; Hansen, Morten

    2016-01-01

    Tumor cells and tumor-infiltrating macrophages produce the chemokine CCL22, which attracts regulatory T cells (Tregs) into the tumor microenvironment, decreasing anticancer immunity. Here, we investigated the possibility of targeting CCL22-expressing cells by activating specific T cells. We...... analyzed the CCL22 protein signal sequence, identifying a human leukocyte antigen A2- (HLA-A2-) restricted peptide epitope, which we then used to stimulate peripheral blood mononuclear cells (PMBCs) to expand populations of CCL22-specific T cells in vitro. T cells recognizing an epitope derived from...... the signal-peptide of CCL22 will recognize CCL22-expressing cells even though CCL22 is secreted out of the cell. CCL22-specific T cells recognized and killed CCL22-expressing cancer cells. Furthermore, CCL22-specific T cells lysed acute monocytic leukemia cells in a CCL22 expression-dependent manner. Using...

  6. SPECIFIC EMISSIONS FROM BIOMASS COMBUSTION

    Directory of Open Access Journals (Sweden)

    Pavel Skopec

    2014-02-01

    Full Text Available This paper deals with determining the specific emissions from the combustion of two kinds of biomass fuels in a small-scale boiler. The tested fuels were pellets made of wood and pellets made of rape plant straw. In order to evaluate the specific emissions, several combustion experiments were carried out using a commercial 25 kW pellet-fired boiler. The specific emissions of CO, SO2 and NOx were evaluated in relation to a unit of burned fuel, a unit of calorific value and a unit of produced heat. The specific emissions were compared with some data acquired from the reference literature, with relatively different results. The differences depend mainly on the procedure used for determining the values, and references provide no information about this. Although some of our experimental results may fit with one of the reference sources, they do not fit with the other. The reliability of the references is therefore disputable.

  7. Site Specific Weed Control Technologies

    DEFF Research Database (Denmark)

    Christensen, Svend; Søgaard, Henning Tangen; Kudsk, Per;

    2009-01-01

    describe the basic parts of site specific weed control technologies, comprising of weed sensing systems, weed management models and precision weed control implements. A review of state-of-the-art technologies shows that several weed sensing systems and precision implements have been developed over the last...... of knowledge about the economic and environmental potential for increasing the resolution of weed control. The integration of site-specific information on weed distribution, weed species composition and density, and the effect on crop yield, is decisive for successful site-specific weed management.   Keywords......Site-specific weed control technologies are defined as machinery or equipment embedded with technologies that detect weeds growing in a crop and, taking into account predefined factors such as economics, takes action to maximise the chances of successfully controlling them. In the article, we...

  8. Eternity Variables to Simulate Specifications

    NARCIS (Netherlands)

    Hesselink, WH; Boiten, EA; Moller, B

    2002-01-01

    Simulation of specifications is introduced as a unification and generalization of refinement mappings, history variables, forward simulations, prophecy variables, and backward simulations. Eternity variables are introduced as a more powerful alternative for prophecy variables and backward

  9. A Domain Specific DSP Processor

    OpenAIRE

    Tell, Eric

    2001-01-01

    This thesis describes the design of a domain specific DSP processor. The thesis is divided into two parts. The first part gives some theoretical background, describes the different steps of the design process (both for DSP processors in general and for this project) and motivates the design decisions made for this processor. The second part is a nearly complete design specification. The intended use of the processor is as a platform for hardware acceleration units. Support for this has howe...

  10. [Specificities of pneumonia in geriatrics].

    Science.gov (United States)

    Pepersack, T

    2014-09-01

    Pneumonia is a major cause of morbidity and mortality leading to a high rate of hospitalization especially in theelderly. It is often a sign of frailty and is associated with a poor prognosis. However, taking into account the geriatric specificities (risk factors, atypical clinical presentations with "geriatric syndromes", ethical debate) using an interdisciplinary and a comprehensive geriatric approach remains an important responsibility of the general practitionner. This article summarizes these specificities and offers interventions targeted on the characteristics of elderly patients.

  11. Specific yield: compilation of specific yields for various materials

    Science.gov (United States)

    Johnson, A.I.

    1967-01-01

    Specific yield is defined as the ratio of (1) the volume of water that a saturated rock or soil will yield by gravity to (2) the total volume of the rock or soft. Specific yield is usually expressed as a percentage. The value is not definitive, because the quantity of water that will drain by gravity depends on variables such as duration of drainage, temperature, mineral composition of the water, and various physical characteristics of the rock or soil under consideration. Values of specific yields nevertheless offer a convenient means by which hydrologists can estimate the water-yielding capacities of earth materials and, as such, are very useful in hydrologic studies. The present report consists mostly of direct or modified quotations from many selected reports that present and evaluate methods for determining specific yield, limitations of those methods, and results of the determinations made on a wide variety of rock and soil materials. Although no particular values are recommended in this report, a table summarizes values of specific yield, and their averages, determined for 10 rock textures. The following is an abstract of the table. [Table

  12. CASL, the Common Algebraic Specification Language

    DEFF Research Database (Denmark)

    Mossakowski, Till; Haxthausen, Anne Elisabeth; Sannella, Donald;

    2008-01-01

    CASL is an expressive specification language that has been designed to supersede many existing algebraic specification languages and provide a standard. CASL consists of several layers, including basic (unstructured) specifications, structured specifications and architectural specifications...

  13. CASL, the Common Algebraic Specification Language

    DEFF Research Database (Denmark)

    Mossakowski, Till; Haxthausen, Anne Elisabeth; Sannella, Donald

    2008-01-01

    CASL is an expressive specification language that has been designed to supersede many existing algebraic specification languages and provide a standard. CASL consists of several layers, including basic (unstructured) specifications, structured specifications and architectural specifications...

  14. NIF optical specifications - the importance of the RMS gradient specification

    Energy Technology Data Exchange (ETDEWEB)

    Auerbach, J M; Cotton, C T; English, R E; Henesian, M A; Hunt J T; Kelly, J H; Lawson, J K; Sacks, J B; Shoup, M J; Trenholme, W H

    1998-07-06

    The performance of the National Ignition Facility (NIF), especially in terms of laser focusability, will be determined by several key factors. One of these key factors is the optical specification for the thousands of large aperture optics that will comprise the 192 beamlines. We have previously reported on the importance of the specification of the power spectral density (PSD) on NIF performance. Recently, we have been studying the importance of long spatial wavelength (>33 mm) phase errors on focusability. We have concluded that the preferred metric for determining the impact of these long spatial wavelength phase errors is the rms phase gradient. In this paper, we outline the overall approach to NIF optical specifications, detail the impact of the rms phase gradient on NIF focusability, discuss its trade-off with the PSD in determining the spot size and review measurements of optics similar to those to be manufactured for NIF.

  15. VHDL Specification Methodology from High-level Specification

    Directory of Open Access Journals (Sweden)

    M. Benmohammed

    2005-01-01

    Full Text Available Design complexity has been increasing exponentially this last decade. In order to cope with such an increase and to keep up designers' productivity, higher level specifications were required. Moreover new synthesis systems, starting with a high level specification, have been developed in order to automate and speed up processor design. This study presents a VHDL specification methodology aimed to extend structured design methodologies to the behavioral level. The goal is to develop VHDL modeling strategies in order to master the design and analysis of large and complex systems. Structured design methodologies are combined with a high-level synthesis system, a VHDL based behavioral synthesis tool, in order to allow hierarchical design and component re-use.

  16. Technical specification for vacuum systems

    Energy Technology Data Exchange (ETDEWEB)

    Khaw, J. (ed.)

    1987-01-01

    The vacuum systems at the Stanford Linear Accelerator Center (SLAC) are primarily of all-metal construction and operate at pressures from 10/sup -5/ to 10/sup -11/ Torr. The primary gas loads during operation result from thermal desorption and beam-induced desorption from the vacuum chamber walls. These desorption rates can be extremely high in the case of hydrocarbons and other contaminants. These specifications place a major emphasis on eliminating contamination sources. The specifications and procedures have been written to insure the cleanliness and vacuum integrity of all SLAC vacuum systems, and to assist personnel involved with SLAC vacuum systems in choosing and designing components that are compatible with existing systems and meet the quality and reliability of SLAC vacuum standards. The specification includes requirements on design, procurement, fabrication, chemical cleaning, clean room practices, welding and brazing, helium leak testing, residual gas analyzer testing, bakeout, venting, and pumpdown. Also appended are specifications regarding acceptable vendors, isopropyl alcohol, bakeable valve cleaning procedure, mechanical engineering safety inspection, notes on synchrotron radiation, and specifications of numerous individual components. (LEW)

  17. 1997 hybrid electric vehicle specifications

    Energy Technology Data Exchange (ETDEWEB)

    Sluder, S.; Larsen, R.; Duoba, M.

    1996-10-01

    The US DOE sponsors Advanced Vehicle Technology competitions to help educate the public and advance new vehicle technologies. For several years, DOE has provided financial and technical support for the American Tour de Sol. This event showcases electric and hybrid electric vehicles in a road rally across portions of the northeastern United States. The specifications contained in this technical memorandum apply to vehicles that will be entered in the 1997 American Tour de Sol. However, the specifications were prepared to be general enough for use by other teams and individuals interested in developing hybrid electric vehicles. The purpose of the specifications is to ensure that the vehicles developed do not present a safety hazard to the teams that build and drive them or to the judges, sponsors, or public who attend the competitions. The specifications are by no means the definitive sources of information on constructing hybrid electric vehicles - as electric and hybrid vehicles technologies advance, so will the standards and practices for their construction. In some cases, the new standards and practices will make portions of these specifications obsolete.

  18. Compiler writing system detail design specification. Volume 1: Language specification

    Science.gov (United States)

    Arthur, W. J.

    1974-01-01

    Construction within the Meta language for both language and target machine specification is reported. The elements of the function language as a meaning and syntax are presented, and the structure of the target language is described which represents the target dependent object text representation of applications programs.

  19. Age‑specific Serum Prostate Specific Antigen Ranges Among ...

    African Journals Online (AJOL)

    Prostate cancer is the most common cancer among men in Nigeria.[1‑4] Currently ..... PSA reference ranges in Chinese men without prostate cancer. Asian J Androl ... of obesity on serum prostate‑specific antigen in Nigerian men. Urol Int 2012 ...

  20. Program Construction by Verifying Specification

    Institute of Scientific and Technical Information of China (English)

    林洪; 陈国良

    1998-01-01

    A program construction method based on Gamma language is proposed.The problem to be solved is specified by first-order predicate logic and a semantic verification program is constructed directly from the specification.Ways for improving efficiency of the program are also studied.The method differs from the one proposed by Manna and Waldinger,where a program is extracted from the proof of the existence of an object meeting the given specification.On the other hand,it also differs from the classical one used for deriving Gamma programs of Banatre and Le metayes,which consists in decomposing the specification into an invariant and a termination conditon.

  1. Specific probiotics or 'fecal transplantation'.

    Science.gov (United States)

    Kruis, Wolfgang

    2012-01-01

    The intestinal ecosystem consists mainly of the enteric flora and to a large extent determines intestinal but also extraintestinal health and disease. General alterations and specific molecular changes of intestinal bacteria cause local as well as systemic immune reactions. Nonantibiotic treatment of the enteric flora has a long tradition and spans a range of different interventions from nutrition to specific probiotics and complete fecal transplantation. When comparing therapy to specific probiotics and fecal transplantation, several aspects need to be considered, like biological consequences, safety and therapeutic evidence. The introduction of probiotics into therapy occurred more than hundred years ago. In contrast, experiences with fecal transplantation are more recent and more limited. Safety issues have not been definitively clarified. Because of the different biological activities of probiotics and fecal transplantation, it can be hypothesized that they may play different roles in the treatment of various diseases. More research is needed before the details, safety and therapeutic effects of bacteriotherapy for IBD become sufficiently clear.

  2. Methylation-Specific PCR Unraveled

    Directory of Open Access Journals (Sweden)

    Sarah Derks

    2004-01-01

    Full Text Available Methylation‐specific PCR (MSP is a simple, quick and cost‐effective method to analyze the DNA methylation status of virtually any group of CpG sites within a CpG island. The technique comprises two parts: (1 sodium bisulfite conversion of unmethylated cytosine's to uracil under conditions whereby methylated cytosines remains unchanged and (2 detection of the bisulfite induced sequence differences by PCR using specific primer sets for both unmethylated and methylated DNA. This review discusses the critical parameters of MSP and presents an overview of the available MSP variants and the (clinical applications.

  3. Portable File Format (PFF) specifications

    Energy Technology Data Exchange (ETDEWEB)

    Dolan, Daniel H. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2015-02-01

    Created at Sandia National Laboratories, the Portable File Format (PFF) allows binary data transfer across computer platforms. Although this capability is supported by many other formats, PFF files are still in use at Sandia, particularly in pulsed power research. This report provides detailed PFF specifications for accessing data without relying on legacy code.

  4. Final Empirical Test Case Specification

    DEFF Research Database (Denmark)

    Kalyanova, Olena; Heiselberg, Per

    This document includes the empirical specification on the IEA task of evaluation building energy simulation computer programs for the Double Skin Facades (DSF) constructions. There are two approaches involved into this procedure, one is the comparative approach and another is the empirical one....

  5. Specific yield, High Plains aquifer

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — This raster data set represents specific-yield ranges in the High Plains aquifer of the United States. The High Plains aquifer underlies 112.6 million acres (176,000...

  6. Adaptive Assessments Using Open Specifications

    Science.gov (United States)

    Leon, Hector Barbosa; Garcia-Penalvo, Francisco J.; Rodriguez-Conde, Maria Jose; Morales, Erla M.; de Pablos, Patricia Ordonez

    2012-01-01

    Evaluation is a key element in formal education processes; it must be constructed in a way that the item questions within help students understand by adapting them to the learning style as well. The focus of the present research work specifically in the convenience to adapt an associated multimedia material in each single question besides the…

  7. [Specificities in children wound healing].

    Science.gov (United States)

    Sanchez, J; Antonicelli, F; Tuton, D; Mazouz Dorval, S; François, C

    2016-10-01

    Children have specific characteristics of wound healing. The aim of this study was to describe the specific clinical characteristics of wounds healing in children and to present the current knowledge on the specific mechanisms with regard to infant age. The tissue insult or injury in fetus can heal without scar, mainly due to reduced granulation tissue associated to diminished or even no inflammatory phase, modified extracellular matrix such as the concentration of hyaluronic acid in amniotic liquid, expression and arrangement of collagen and tenascin. Thickness of children skin is a serious negative factor in case of trauma, whereas poor co-morbidities and efficient growth tissue mechanisms are beneficial to good evolution, even in cases of extensive damage and loss of tissue. The subsequent tissue mechanical forces, wound healing during childhood, spanning from the age of 2 until the end of puberty, is associated with more hypertrophic scars, both in duration and in intensity. Consequently, unnecessary surgery has to be avoided during this period when possible, and children with abnormal or pathologic wound healing should benefit from complementary treatments (hydration, massage, brace, silicone, hydrotherapy…), which represent efficient factors to minimize tissue scarring. After wound healing, the growth body rate can be responsible for specific complications, such as contractures, alopecia, and scar intussusceptions. Its evolutionary character implies the need of an attentive follow-up until adult age. Psychologic repercussions, as a consequence of pathologic scars, must be prevented and investigated by the surgeon.

  8. CONTRACT DOCUMENTS AND PERFORMANCE SPECIFICATIONS.

    Science.gov (United States)

    First California Commission on School Construction Systems, Palo Alto.

    A SET OF EDUCATIONAL SPECIFICATION DRAFTED BY THE FIRST CALIFORNIA COMMISSION ON SCHOOL CONSTRUCTION SYSTEMS GIVES INFORMATION ON BIDDING PROCEDURES, A DESCRIPTION OF THE CURRENT CONSTRUCTION PROGRAM, PROCEDURES FOR SUBMITTING A PROPOSAL, DATA AND CONDITIONS RELATED TO THE DEVELOPMENT PHASE OF THE PROJECT, COMPONENT CONTRACTS, AND GENERAL…

  9. Priming the Query Specification Process.

    Science.gov (United States)

    Toms, Elaine G.; Freund, Luanne

    2003-01-01

    Tests the use of questions as a technique in the query specification process. Using a within-subjects design, 48 people interacted with a modified Google interface to solve four information problems in four domains. Half the tasks were entered as typical keyword queries, and half as questions or statements. Results suggest the typical search box…

  10. Behaviour specification in database interoperation

    NARCIS (Netherlands)

    Vermeer, W.W.M.; Apers, Peter M.G.

    We discuss the impact of locally implemented behaviour in a federation of object-oriented databases. In particular, given a specification of an integrated view of a number of component databases, we discuss the process of determining the global methods that are implicitly implemented by a given set

  11. Domain specific MT in use

    DEFF Research Database (Denmark)

    Offersgaard, Lene; Povlsen, Claus; Almsten, Lisbeth Kjeldgaard

    2008-01-01

    The paper focuses on domain specific use of MT with a special focus on SMT in the workflow of a Language Service Provider (LSP). We report on the feedback of post-editors using fluency/adequacy evaluation and the evaluation metric ’Usability’, understood in this context as where users on a three ...

  12. α-Fetoprotein promoter-driven Cre/LoxP-switched RNA interference for hepatocellular carcinoma tissue-specific target therapy.

    Directory of Open Access Journals (Sweden)

    Yuan-Fei Peng

    Full Text Available BACKGROUND: RNA interference (RNAi has recently emerged as a potential treatment modality for hepatocellular carcinoma (HCC therapy, but the lack of cellular targets and sustained efficacy limits its application. The purpose of this study is to develop an HCC tissue-specific RNAi system and investigate its possibility for HCC treatment. METHODS: Two different HCC-specific RNAi systems in which therapeutic miRNA or shRNA against target gene (Beclin 1 was directly or indirectly driven by alpha-fetoprotein promoter (AFP-miRNA and AFP-Cre/LoxP-shRNA were constructed. Human HCC cell lines (HepG2, Hep3B and HCCLM3 and non-HCC cell lines (L-02, Hela and SW1116 were infected with the systems. The effectiveness and tissue-specificity of the systems were examined by Q-PCR and western blot analysis. The efficacy of the systems was further tested in mouse model of HCC by intravenous or intratumoral administration. The feasibility of the system for HCC treatment was evaluated by applying the system as adjuvant therapy to enhance sorafenib treatment. An AFP-Cre/LoxP-shRNA system targeting Atg5 gene (AFP-Cre/LoxP-shRNA-Atg5 was constructed and its efficacy in sensitizing HCC cells (MHCC97L/PLC to sorafenib treatment was examined by apoptosis assay in vitro and tumorigenesis assay in vivo. RESULTS: The AFP-miRNA system could silence target gene (Beclin 1 but required a high titer which was lethal to target cells. The AFP-Cre/LoxP-shRNA system could efficiently knockdown target gene while maintain high HCC specificity. Intratumoral injection of the AFP-Cre/LoxP-shRNA system could efficiently silence target gene (Beclin 1 in vivo while intravenous administration could not. The AFP-Cre/LoxP-shRNA system target Atg5 gene could significantly sensitize MHCC97L/PLC cells to sorafenib-induced apoptosis in vitro and tumor growth suppression in vivo. CONCLUSIONS: An efficient HCC tissue-specific RNAi system (AFP-Cre/LoxP-shRNA was successfully established. The system

  13. Epididymis-specific lipocalin promoters

    Institute of Scientific and Technical Information of China (English)

    Kichiya Suzuki; Xiuping Yu; Pierre Chaurand; Yoshihiko Araki; Jean-Jacques Lareyre; Richard M. Caprioli; Marie-Claire Orgebin-Crist; Robert J. Matusik

    2007-01-01

    Our goal is to decipher which DNA sequences are required for tissue-specific expression of epididymal genes. At least 6 epididymis-specific lipocalin genes are known. These are differently regulated and regionalized in the epididymis.Lipocalin 5 (Lcn5 or mE-RABP) and Lipocalin 8 (Lcn8 or mEP17) are homologous genes belonging to the epididymis-specific lipocalin gene cluster. Both the 5 kb promoter fragment of the Lcn5 gene and the 5.3 kb promoter fragment of the Lcn8 gene can direct transgene expression in the epididymis (Lcn5 to the distal caput and Lcn8 to the initial segment), indicating that these promoter fragments contain important cis-regulatory element(s) for epididymisspecific gene expression. To define further the fragments regulating gene expression, the Lcn5 promoter was examined in transgenic mice and immortalized epididymal cell lines. After serial deletion, the 1.8 kb promoter fragment of the Lcn5 gene was sufficient for tissue-specific and region-specific gene expression in transgenic mice. Transient transfection analysis revealed that a transcription factor forkhead box A2 (Foxa2) interacts with androgen receptor and binds to the 100 bp fragment of the Lcn5 promoter between 1.2 kb and 1.3 kb and that Foxa2 expression inhibitsandrogen-dependent induction of the Lcn5 promoter activity. Immunohistochemistry indicated a restricted expression of Foxa2 in the epididymis where endogenous Lcn5 gene expression is suppressed and that the Foxa2 inhibition of the Lcn5 promoter is consistent with the lack of expression of Lcn5 in the corpus and cauda. Our approach provides a basic strategy for further analysis of the epididymal lipocalin gene regulation and flexible control of epididymal function.

  14. Synthetic Pre-miRNA-Based shRNA as Potent RNAi Triggers

    Directory of Open Access Journals (Sweden)

    Kazuya Terasawa

    2011-01-01

    Full Text Available RNA interference (RNAi is a powerful tool for studying gene function owing to the ease with which it can selectively silence genes of interest, and it has also attracted attention because of its potential for therapeutic applications. Chemically synthesized small interfering RNAs (siRNAs and DNA vector-based short hairpin RNAs (shRNAs are now widely used as RNAi triggers. In contrast to expressed shRNAs, the use of synthetic shRNAs is limited. Here we designed shRNAs modeled on a precursor microRNA (pre-miRNA and evaluated their biological activity. We demonstrated that chemically synthetic pre-miRNA-based shRNAs have more potent RNAi activity than their corresponding siRNAs and found that their antisense strands are more efficiently incorporated into the RNA-induced silencing complex. Although greater off-target effects and interferon responses were induced by shRNAs than by their corresponding siRNAs, these effects could be overcome by simply using a lower concentration or by optimizing and chemically modifying shRNAs similar to synthetic siRNAs. These are challenges for the future.

  15. Inducible shRNA expression for application in a prostate cancer mouse model.

    Science.gov (United States)

    Czauderna, Frank; Santel, Ansgar; Hinz, Michael; Fechtner, Melanie; Durieux, Birgit; Fisch, Gerald; Leenders, Frauke; Arnold, Wolfgang; Giese, Klaus; Klippel, Anke; Kaufmann, Jörg

    2003-11-01

    RNA interference (RNAi) is a powerful tool to induce loss-of-function phenotypes by inhibiting gene expression post-transcriptionally. Synthetic short interfering RNAs (siRNAs) as well as vector-based siRNA expression systems have been used successfully to silence gene expression in a variety of biological systems. We describe the development of an inducible siRNA expression system that is based on the tetracycline repressor and eukaryotic RNA polymerase III promoters (U6 and 7SK). For proof of concept we selectively inhibited expression of two catalytic subunits of the phosphatidylinositol 3-kinase (PI 3-kinase), p110alpha and p110beta, by using vector-derived short hairpin RNAs (shRNAs). Stable pools of human prostate cancer cells (PC-3) exhibiting reduced levels of both PI 3-kinase catalytic subunits due to the expression of corresponding shRNAs in an inducible fashion were established and analyzed for their invasive potential in vitro as well as in an orthotopic metastatic mouse model. This inducible system for RNAi allows an unbiased and comparable analysis of loss-of-function phenotypes by comparing selected isogenic cell populations on the induced and non-induced level. In addition, conditional RNAi allows the study of essential and multifunctional genes involved in complex biological processes by preventing inhibitory and compensatory effects caused by constitutive knockdown.

  16. SHRNA SILENCING OF AS3MT EXPRESSION MINIMIZES ARSENIC METHYLATION CAPACITY OF HEPG2 CELLS

    Science.gov (United States)

    Several methyltransferases have been shown to catalyze the oxidative methylation of inorganic arsenic (iAs) in mammalian species. However, the relative contributions of these enzymes to the overall capacity of cells to methylate iAs have not been characterized. Arsenic (+3 oxidat...

  17. Defective neuronal migration and inhibition of bipolar to multipolar transition of migrating neural cells by Mesoderm-Specific Transcript, Mest, in the developing mouse neocortex.

    Science.gov (United States)

    Ji, Liting; Bishayee, Kausik; Sadra, Ali; Choi, Seunghyuk; Choi, Wooyul; Moon, Sungho; Jho, Eek-Hoon; Huh, Sung-Oh

    2017-07-04

    Brain developmental disorders such as lissencephaly can result from faulty neuronal migration and differentiation during the formation of the mammalian neocortex. The cerebral cortex is a modular structure, where developmentally, newborn neurons are generated as a neuro-epithelial sheet and subsequently differentiate, migrate and organize into their final positions in the cerebral cortical plate via a process involving both tangential and radial migration. The specific role of Mest, an imprinted gene, in neuronal migration has not been previously studied. In this work, we reduced expression of Mest with in utero electroporation of neuronal progenitors in the developing embryonic mouse neocortex. Reduction of Mest levels by shRNA significantly reduced the number of neurons migrating to the cortical plate. Also, Mest-knockdown disrupted the transition of bipolar neurons into multipolar neurons migrating out of the sub-ventricular zone region. The migrating neurons also adopted a more tangential migration pattern upon knockdown of the Mest message, losing their potential to attach to radial glia cells, required for radial migration. The differentiation and migration properties of neurons via Wnt-Akt signaling were affected by Mest changes. In addition, miR-335, encoded in a Mest gene intron, was identified as being responsible for blocking the default tangential migration of the neurons. Our results suggest that Mest and its intron product, miR-335, play important roles in neuronal migration with Mest regulating the morphological transition of primary neurons required in the formation of the mammalian neocortex. Copyright © 2017 IBRO. Published by Elsevier Ltd. All rights reserved.

  18. New alternatively spliced variant of prostate-specific membrane antigen PSM-E suppresses the proliferation, migration and invasiveness of prostate cancer cells.

    Science.gov (United States)

    Cao, Kai-Yuan; Xu, Lin; Zhang, Ding-Mei; Zhang, Xiao-Ming; Zhang, Tian; He, Xia; Wang, Zhu; Feng, Fa-Shen; Qiu, Shao-Peng; Shen, Guan-Xin

    2012-06-01

    PSM-E is a newly discovered alternatively spliced variant of prostate-specific membrane antigen (PSMA). In the current study, its role on the proliferation, invasiveness and migration in prostate cancer cell lines was analyzed. PSM-E and PSMA (as a comparison) eukaryotic expression vectors pcDNA3.0/PSM-E and pcDNA3.0/PSMA were constructed, validated by RT-PCR and Western blotting, and PSMA/PSM-E overexpression PC-3 cell models were built. Gene interference was used to block PSMA and the expression of its splice variants in LNCap cells. Three shRNA fragments were synthesized against PSMA, cloned into the vector pSilencer 2.1-U6-neo, their interference effect was evaluated by RT-PCR and Western blotting, and pSilencer 2.1-U6-neo‑shRNA3 (named p‑shRNA3) was chosen in further analyses. Growth curves were drawn to observe the proliferation change, which showed that PSM-E had the potential to suppress proliferation (PPSM-E interfering LNCap cells (P>0.05). Cross-river test showed that the migration speeds of PSM-E/PC-3 and PSMA/PC-3 were both significantly slower than the vector negative control, and faster in p-shRNA3 interfering LNCap cells compared with its vector negative control (PPSM-E/PC-3 and PSMA/PC-3 (P>0.05). Transwell assay showed that the invasive cells of both PSMA/PC-3 and PSM-E/PC-3 were fewer compared to the vector negative control (PPSM-E was weaker than PSMA (PPSM-E could suppress proliferation, migration and invasiveness of prostate cancer cells. Its suppression effect on cell proliferation is stronger compared to PSMA and the suppression effect on invasiveness is weaker than that of PSMA.

  19. Tissue Specific Labeling in Proteomics

    Directory of Open Access Journals (Sweden)

    Evelyn Ramberger

    2017-07-01

    Full Text Available Mass spectrometry-based proteomics is a powerful tool for identifying and quantifying proteins in biological samples. While it is routinely used for the characterization of simple cell line systems, the analysis of the cell specific proteome in multicellular organisms and tissues poses a significant challenge. Isolating a subset of cells from tissues requires mechanical and biochemical separation or sorting, a process which can alter cellular signaling, and thus, the composition of the proteome. Recently, several approaches for cell selective labeling of proteins, that include bioorthogonal amino acids, biotinylating enzymes, and genetic tools, have been developed. These tools facilitate the selective labeling of proteins, their interactome, or of specific cell types within a tissue or an organism, while avoiding the difficult and contamination-prone biochemical separation of cells from the tissue. In this review, we give an overview of existing techniques and their application in cell culture models and whole animals.

  20. A development calculus for specifications

    Institute of Scientific and Technical Information of China (English)

    李未

    2003-01-01

    A first order inference system, named R-calculus, is defined to develop the specifications.This system intends to eliminate the laws which are not consistent with users' requirements. TheR-calculus consists of the structural rules, an axiom, a cut rule, and the rules for logical connectives.Some examples are given to demonstrate the usage of the R-calculus. Furthermore, the propertiesregarding reachability and completeness of the R-calculus are formally defined and proved.

  1. [Quality specification of Curcumae Radix].

    Science.gov (United States)

    Zhu, Jingjing; Zhang, Qingzhe; Wang, Zhimin; Zhang, Qiwei

    2010-08-01

    To establish and revise the quality specification of Curcumae Radix. TLC identification was carried out by the method in Appendix VI B in Chinese pharmacopoeia (2005th Edition) and the contents of volatile oil was determined by the method describing in Appendix XD. Twenty-four Samples from four different curcuma species were performed TLC qualitative analysis and volatile determination, respectively. The established method can be used for the quality control of Curcumae Radix.

  2. Rapid Estimation of Specific Impulse

    Directory of Open Access Journals (Sweden)

    V. K. Bhat

    1988-01-01

    Full Text Available Specific impulse (Isp is one of the important criteria for propellant characterisation. In the present paper a semiempirical approach has been suggested to calculate Isp of composite modified doublebase(CMDB propellant formulations by utilizing calorimetric value(Cal-Val of the propellant composition. The Cal-Val of aluminium has been computed on the basis of oxygen balance of the propellant composition. The validity of the new approach has been demonstratedby comparing predicted values witli actual results.

  3. Lack of specificity of commercially available antisera: better specifications needed.

    Science.gov (United States)

    Pradidarcheep, Wisuit; Labruyère, Wil T; Dabhoiwala, Noshir F; Lamers, Wouter H

    2008-12-01

    The ideal antiserum for immunohistochemical (IHC) applications contains monospecific high-affinity antibodies with little nonspecific adherence to sections. Many commercially available antibodies are "affinity" purified, but it is unknown if they meet "hard" specificity criteria, such as absence of staining in tissues genetically deficient for the antigen or a staining pattern that is identical to that of an antibody raised against a different epitope on the same protein. Reviewers, therefore, often require additional characterization. Although the affinity-purified antibodies used in our study on the distribution of muscarinic receptors produced selective staining patterns on sections, few passed the preabsorption test, and none produced bands of the anticipated size on Western blots. More importantly, none showed a difference in staining pattern on sections or Western blots between wild-type and knockout mice. Because these antibodies were used in most studies published thus far, our findings cast doubts on the validity of the extant body of morphological knowledge of the whole family of muscarinic receptors. We formulate requirements that antibody-specification data sheets should meet and propose that journals for which IHC is a core technique facilitate consumer rating of antibodies. "Certified" antibodies could avoid fruitless and costly validation assays and should become the standard of commercial suppliers.

  4. How specific is specific phobia? Different neural response patterns in two subtypes of specific phobia.

    Science.gov (United States)

    Lueken, Ulrike; Kruschwitz, Johann Daniel; Muehlhan, Markus; Siegert, Jens; Hoyer, Jürgen; Wittchen, Hans-Ulrich

    2011-05-01

    Specific phobia of the animal subtype has been employed as a model disorder exploring the neurocircuitry of anxiety disorders, but evidence is lacking whether the detected neural response pattern accounts for all animal subtypes, nor across other phobia subtypes. The present study aimed at directly comparing two subtypes of specific phobia: snake phobia (SP) representing the animal, and dental phobia (DP) representing the blood-injection-injury subtype. Using functional magnetic resonance imaging (fMRI), brain activation and skin conductance was measured during phobogenic video stimulation in 12 DP, 12 SP, and 17 healthy controls. For SP, the previously described activation of fear circuitry structures encompassing the insula, anterior cingulate cortex and thalamus could be replicated and was furthermore associated with autonomic arousal. In contrast, DP showed circumscribed activation of the prefrontal and orbitofrontal cortex (PFC/OFC) when directly compared to SP, being dissociated from autonomic arousal. Results provide preliminary evidence for the idea that snake and dental phobia are characterized by distinct underlying neural systems during sustained emotional processing with evaluation processes in DP being controlled by orbitofrontal areas, whereas phobogenic reactions in SP are primarily guided by limbic and paralimbic structures. Findings support the current diagnostic classification conventions, separating distinct subtypes in DSM-IV-TR. They highlight that caution might be warranted though for generalizing findings derived from animal phobia to other phobic and anxiety disorders. If replicated, results could contribute to a better understanding of underlying neurobiological mechanisms of specific phobia and their respective classification.

  5. Advances in bacterial specific imaging

    Energy Technology Data Exchange (ETDEWEB)

    Wareham, David; Das, Satya [St Bartholomew' s Hospital, London (United Kingdom). Dept. of Medical Microbiology; London Univ. (United Kingdom). Queen Mary' s School of Medicine and Dentistry. Inst. of Cell and Molecular Science

    2005-10-15

    Nuclear medicine is a powerful diagnostic technique able to detect inflammatory foci in human disease. A wide range of agents have been evaluated for their ability to distinguish lesions due to microbial infection from those due to sterile inflammation. Advances continue to be made on the use of radiolabelled antibiotics which as well as being highly specific in the diagnosis of infection may be useful in monitoring the treatment and course of disease. Here we provide an update on in-vitro and clinical studies with a number of established and novel radiopharmaceuticals. (author)

  6. Vibrio chromosome-specific families

    DEFF Research Database (Denmark)

    Lukjancenko, Oksana; Ussery, David

    2014-01-01

    families, 1169 and 153 are uniquely found in chromosomes 1 and 2, respectively. Gene ontology (GO) terms for each of the protein families were determined, and the different sets for each chromosome were compared. A total of 363 different "Molecular Function" GO categories were found for chromosome 1......We have compared chromosome-specific genes in a set of 18 finished Vibrio genomes, and, in addition, also calculated the pan- and core-genomes from a data set of more than 250 draft Vibrio genome sequences. These genomes come from 9 known species and 2 unknown species. Within the finished...

  7. IntelliCable Interface Specification

    Science.gov (United States)

    2016-07-14

    manager on how to use this. Some will factor the priority into account and always consume the highest priority while others...Battlefield Air Operations power manager uses a 10 pin Glenair Mighty Mouse connector with default keying . Pin 9 is specified as the IntelliCable pin. A BAO...standard Soldier Power Manager uses a 7 pin Glenair Mighty Mouse Connector, using alternative keying distinct from the Land Warrior battery specification, and the Li-145 battery. Pin 3 is defined as the IntelliCable pin.

  8. [Quality specification of Perillae Fructus].

    Science.gov (United States)

    Gu, Lihua; Lin, Chen; Wu, Tao; Chou, Guixin; Wang, Zhengtao

    2010-08-01

    To improve the quality control specification of Perillae Fructus, the identification methods and assay were developed. Rosmarinic acid, luteolin and apigenin in the sample were identified by TLC. The content of rosmarinic acid was determined by HPLC. The linear calibration curve of rosmarinic acid was obtained in the ranges of 19.4-194.2 g x L(-1) (R2 = 0.9999). The arerage coveriy (n=9) for the assay was 99.8% (RSD 3.6%). The established methods are accuracy, sensitivity and reproducible, and can be used for the quality control of Perillae Fructus.

  9. Project X functional requirements specification

    CERN Document Server

    Holmes, S D; Kephart, R; Kerby, J; Kourbanis, I; Lebedev, V; Mishra, S; Nagaitsev, S; Solyak, N; Tschirhart, R

    2012-01-01

    Project X is a multi-megawatt proton facility being developed to support a world-leading program in Intensity Frontier physics at Fermilab. The facility is designed to support programs in elementary particle and nuclear physics, with possible applications to nuclear energy research. A Functional Requirements Specification has been developed in order to establish performance criteria for the Project X complex in support of these multiple missions, and to assure that the facility is designed with sufficient upgrade capability to provide U.S. leadership for many decades to come. This paper will briefly review the previously described Functional Requirements, and then discuss their recent evolution.

  10. Specifications of EA-BRACHI

    Energy Technology Data Exchange (ETDEWEB)

    Taguchi, Mitsumasa; Namba, Hideki; Aoki, Yasushi; Watanabe, Ritsuko; Matsumoto, Yuuichi; Watanabe, Hiroshi [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    1996-11-01

    EA-BRACHI (Experimental Apparatus for Basic study of Radiation Chemistry with Heavy Ions) has been constructed at a vertical port of AVF cyclotron in TIARA (Takasaki Ion Accelerators for Advanced Radiation Application) facility. This apparatus is used for measurement of the spatial distribution of energy deposited around the ion`s path, and the reaction of radicals and other intermediates produced by heavy ion irradiation in solid and liquid samples. This report detailed specifications and performance of this apparatus, including operational procedures and some basic data. (author)

  11. Central Solenoid Insert Technical Specification

    Energy Technology Data Exchange (ETDEWEB)

    Martovetsky, Nicolai N [ORNL; Smirnov, Alexandre [ORNL

    2011-09-01

    The US ITER Project Office (USIPO) is responsible for the ITER central solenoid (CS) contribution to the ITER project. The Central Solenoid Insert (CSI) project will allow ITER validation the appropriate lengths of the conductors to be used in the full-scale CS coils under relevant conditions. The ITER Program plans to build and test a CSI to verify the performance of the CS conductor. The CSI is a one-layer solenoid with an inner diameter of 1.48 m and a height of 4.45 m between electric terminal ends. The coil weight with the terminals is approximately 820 kg without insulation. The major goal of the CSI is to measure the temperature margin of the CS under the ITER direct current (DC) operating conditions, including determining sensitivity to load cycles. Performance of the joints, ramp rate sensitivity, and stability against thermal or electromagnetic disturbances, electrical insulation, losses, and instrumentation are addressed separately and therefore are not major goals in this project. However, losses and joint performance will be tested during the CSI testing campaign. The USIPO will build the CSI that will be tested at the Central Solenoid Model Coil (CSMC) Test Facility at the Japan Atomic Energy Agency (JAEA), Naka, Japan. The industrial vendors (the Suppliers) will report to the USIPO (the Company). All approvals to proceed will be issued by the Company, which in some cases, as specified in this document, will also require the approval of the ITER Organization. Responsibilities and obligations will be covered by respective contracts between the USIPO, called Company interchangeably, and the industrial Prime Contractors, called Suppliers. Different stages of work may be performed by more than one Prime Contractor, as described in this specification. Technical requirements of the contract between the Company and the Prime Contractor will be covered by the Fabrication Specifications developed by the Prime Contractor based on this document and approved by

  12. Substrate Specificity of Prostate-Specific Membrane Antigen

    Science.gov (United States)

    Anderson, Marc O.; Wu, Lisa Y.; Santiago, Nicholas M.; Moser, Jamie M.; Rowley, Jennifer A.; Bolstad, Erin S. D.; Berkman, Clifford E.

    2007-01-01

    A series of putative dipeptide substrates of prostate specific membrane antigen (PSMA) was prepared that explored α- and β/γ-linked acidic residues at the P1 position and various chromophores at the P2 position, while keeping the P1’ residue constant as L-Glu. Four chromophores were examined, including 4-phenylazobenzoyl, 1-pyrenebutyrl, 9-anthracenylcarboxyl-γ-aminobutyrl, and 4-nitrophenylbutyryl. When evaluating these chromophores, it was found that a substrate containing 4-phenylazobenzoyl at the P2 position was consumed most efficiently. Substitution at the P1 position with acidic residues showed that only γ-linked L-Glu and D-Glu were recognized by the enzyme, with the former being more readily proteolyzed. Lastly, binding modes of endogenous substrates and our best synthetic substrate (4-phenylazobenzoyl-Glu-γ-Glu) were proposed by computational docking studies into an X-ray crystal structure of the PSMA extracellular domain. PMID:17764959

  13. Flight Guidance System Requirements Specification

    Science.gov (United States)

    Miller, Steven P.; Tribble, Alan C.; Carlson, Timothy M.; Danielson, Eric J.

    2003-01-01

    This report describes a requirements specification written in the RSML-e language for the mode logic of a Flight Guidance System of a typical regional jet aircraft. This model was created as one of the first steps in a five-year project sponsored by the NASA Langley Research Center, Rockwell Collins Inc., and the Critical Systems Research Group of the University of Minnesota to develop new methods and tools to improve the safety of avionics designs. This model will be used to demonstrate the application of a variety of methods and techniques, including safety analysis of system and subsystem requirements, verification of key properties using theorem provers and model checkers, identification of potential sources mode confusion in system designs, partitioning of applications based on the criticality of system hazards, and autogeneration of avionics quality code. While this model is representative of the mode logic of a typical regional jet aircraft, it does not describe an actual or planned product. Several aspects of a full Flight Guidance System, such as recovery from failed sensors, have been omitted, and no claims are made regarding the accuracy or completeness of this specification.

  14. [Pharmacological gender-specific medicine].

    Science.gov (United States)

    Ueno, Koichi; Negishi, Etsuko

    2006-01-01

    In recent years, the concept of Gender-specific Medicine has become more widespread in Japan. We need to understand gender differences in the use patterns of prescription drugs for proper use of medicines. We therefore investigated gender differences in the use of prescription drugs using the data of 9 hospitals in Japan. The data on prescription drugs were extracted from the drug ordering system for a month from March 1 to 31, 2003. We analyzed the data according to sex and age. As a result, we found that the prescription frequency of central nervous system drugs and Kampo medications for females was higher than that for males. There was also the same trend for hormone medicines and vitamins. On the other hand, the prescription frequency of cardiovascular drugs for males was higher than that for females. Moreover, there was the same trend for unclassified metabolic drugs such as arthrifuge. As a result of detailed analysis by age, a correlation between age-specificity of prescription drugs and gender difference of diseases was suggested. Such information has not been investigated in Japan. Since these results are thought to be useful, we will perform a detailed examination and accumulate evidence for their future exploitation in drug therapy.

  15. Arithmetic Memory Is Modality Specific.

    Directory of Open Access Journals (Sweden)

    Timothy Myers

    Full Text Available In regards to numerical cognition and working memory, it is an open question as to whether numbers are stored into and retrieved from a central abstract representation or from separate notation-specific representations. This study seeks to help answer this by utilizing the numeral modality effect (NME in three experiments to explore how numbers are processed by the human brain. The participants were presented with numbers (1-9 as either Arabic digits or written number words (Arabic digits and dot matrices in Experiment 2 at the first (S1 and second (S2 stimuli. The participant's task was to add the first two stimuli together and verify whether the answer (S3, presented simultaneously with S2, was correct. We hypothesized that if reaction time (RT at S2/S3 depends on the modality of S1 then numbers are retrieved from modality specific memory stores. Indeed, RT depended on the modality of S1 whenever S2 was an Arabic digit which argues against the concept of numbers being stored and retrieved from a central, abstract representation.

  16. BREAKTHROUGH INDEX AND SPECIFIC DEPOSIT

    Directory of Open Access Journals (Sweden)

    Awatif Soaded Al-Saqqar

    2013-05-01

    Full Text Available The dual filter was tested in this study to improve the performance of the filtration process in water treatment plants. Porcelanite rocks were selected to be the dual media with sand in the experimental work. The work required installing a pilot filtration unit in the location of the filters in one of the water treatment plants. The pilot filtration consists of three plastic column filters, acting parallel and simultaneously. The first contains 70 cm sand, the second and third were dual filters (porcelanite with sand of different types. The dual media was tested at different filtration rates (5, 7.5, 10, and 15 m/hr. The results showed that the dual filters had better performance than sand filters and reduced the specific deposit (σ and the breakthrough index (BI. In the dual filters the specific deposit was about (16 to 65 % less than in sand filters and the breakthrough index (BI was specified weak for rates 5 and 7 m/hr, light at 10 m/hr, and medium at 15 m/hr.

  17. Distribution-Specific Agnostic Boosting

    CERN Document Server

    Feldman, Vitaly

    2009-01-01

    We consider the problem of boosting the accuracy of weak learning algorithms in the agnostic learning framework of Haussler (1992) and Kearns et al. (1992). Known algorithms for this problem (Ben-David et al., 2001; Gavinsky, 2002; Kalai et al., 2008) follow the same strategy as boosting algorithms in the PAC model: the weak learner is executed on the same target function but over different distributions on the domain. We demonstrate boosting algorithms for the agnostic learning framework that only modify the distribution on the labels of the points (or, equivalently, modify the target function). This allows boosting a distribution-specific weak agnostic learner to a strong agnostic learner with respect to the same distribution. When applied to the weak agnostic parity learning algorithm of Goldreich and Levin (1989) our algorithm yields a simple PAC learning algorithm for DNF and an agnostic learning algorithm for decision trees over the uniform distribution using membership queries. These results substantia...

  18. The Specificity of Human Aesthetics

    DEFF Research Database (Denmark)

    Bouchet, Dominique

    2015-01-01

    Today, humankind is being reintroduced within nature. Humankind is no longer presented as supernatural to nature. There is no longer a clear divide between animal and human. What makes us human is neither culture, nor language nor labor, nor art, but the degree of complexity those products and ca...... creativity. Art is fundamentally about shaping visions and very closely related to this specific human ability to make projects. Especially in times of crisis, when the old sources of meaning hardly can make sense in a new context, imagination must be used in order to survive......., and the craving to make sense of all kinds of relations that characterizes humans. Being able to produce artworks as external representations aimed at “the other,” made of internal representations is thus essential to humanity. Human aesthetics is not so much an issue of beauty but of imagination and social...

  19. MATISSE: specifications and expected performances

    CERN Document Server

    Matter, A; Petrov, R G; Berio, P; Robbe-Dubois, S; Lopez, B; Antonelli, P; Allouche, F; Cruzalebes, P; Millour, F; Bazin, G; Bourgès, L

    2016-01-01

    MATISSE (Multi AperTure mid-Infrared SpectroScopic Experiment) is the next generation spectro-interferometer at the European Southern Observatory VLTI operating in the spectral bands L, M and N, and combining four beams from the unit and auxiliary telescopes. MATISSE is now fully integrated at the Observatoire de la C\\^ote d'Azur in Nice (France), and has entered very recently its testing phase in laboratory. This paper summarizes the equations describing the MATISSE signal and the associated sources of noise. The specifications and the expected performances of the instrument are then evaluated taking into account the current characteristics of the instrument and the VLTI infrastructure, including transmission and contrast degradation budgets. In addition, we present the different MATISSE simulation tools that will be made available to the future users.

  20. Specifications for gas treatment apparatus

    Energy Technology Data Exchange (ETDEWEB)

    Teraoka, Yuden; Yoshigoe, Akitaka [Japan Atomic Energy Research Inst., Kansai Research Establishment, Synchrotron Radiation Research Center, Mikazuki, Hyogo (Japan)

    2001-03-01

    A surface reaction analysis apparatus was installed as an experimental end-station for the study of surface chemistry at the JAERI soft x-ray beamline in the SPring-8. The surface reaction analysis apparatus is devoted to the study of reaction mechanisms on solid surfaces with incident gas molecules. It is necessary to supply reagent gases to the apparatus. Chlorine and metal organic molecules will be used in the apparatus as well as oxygen molecules. Since the chlorine is corrosive and virulent, the metal organic molecules are flammable, the satiety treatments and the removal of virulence from the exhaust gas are needed. This gas supply and exhaust system is mainly composed of a cylinder cabinet, a gas mixer, a virulence removal cell and an urgent virulence removal apparatus. The former three devices are considered to take a countermeasure against virulent gas leak. The detail specifications concerning the gas supply and exhaust system are described in this report. (author)

  1. Genetic specificity of face recognition.

    Science.gov (United States)

    Shakeshaft, Nicholas G; Plomin, Robert

    2015-10-13

    Specific cognitive abilities in diverse domains are typically found to be highly heritable and substantially correlated with general cognitive ability (g), both phenotypically and genetically. Recent twin studies have found the ability to memorize and recognize faces to be an exception, being similarly heritable but phenotypically substantially uncorrelated both with g and with general object recognition. However, the genetic relationships between face recognition and other abilities (the extent to which they share a common genetic etiology) cannot be determined from phenotypic associations. In this, to our knowledge, first study of the genetic associations between face recognition and other domains, 2,000 18- and 19-year-old United Kingdom twins completed tests assessing their face recognition, object recognition, and general cognitive abilities. Results confirmed the substantial heritability of face recognition (61%), and multivariate genetic analyses found that most of this genetic influence is unique and not shared with other cognitive abilities.

  2. Specific Carbohydrate Diet: Does It Work?

    Science.gov (United States)

    ... Specific Carbohydrate Diet (SCD) Go Back The Specific Carbohydrate Diet (SCD) Email Print + Share There is no ... diet that has received attention is the Specific Carbohydrate Diet. This diet limits poorly digestible carbohydrates to ...

  3. Eternity Variables to Prove Simulation of Specifications

    NARCIS (Netherlands)

    Hesselink, Wim H.

    2005-01-01

    Simulations of specifications are introduced as a unification and generalization of refinement mappings, history variables, forward simulations, prophecy variables, and backward simulations. A specification implements another specification if and only if there is a simulation from the first one to

  4. FY97 ICCS prototype specification

    Energy Technology Data Exchange (ETDEWEB)

    Woodruff, J.

    1997-02-20

    The ICCS software team will implement and test two iterations of their software product during FY97. The first of these iterations will concentrate on construction of selected framework components; the subsequent iteration will extend the product and perform measurements of performance based on emulated FEP devices. This document specifies the products to be delivered in that first prototype and projects the direction that the second prototype will take. Detailed specification of the later iteration will be written when the results of the first iteration are complete. The selection of frameworks to be implemented early is made on a basis of risk analysis from the point of view of future development in the ICCS project. The prototype will address risks in integration of object- oriented components, in refining our development process, and in emulation testing for FEP devices. This document is a specification that identifies products and processes to undertake for resolving these risks. The goals of this activity are to exercise our development process at a modest scale and to probe our architecture plan for fundamental limits and failure modes. The product of the iterations will be the framework software which will be useful in future ICCS code. Thus the FY97 products are intended for internal usage by the ICCS team and for demonstration to the FEP software developers of the strategy for integrating supervisory software with FEP computers. This will be the first of several expected iterations of the software development process and the performance measurements that ICCS will demonstrate, intended to support confidence in our ability to meet project RAM goals. The design of the application software is being carried out in a separate WBS 1.5.2 activity. The design activity has as its FY97 product a series of Software Design Documents that will specify the functionality of the controls software of ICCS. During the testing of this year`s prototypes, the application

  5. Specificities of sustainable tourism planning

    Directory of Open Access Journals (Sweden)

    Jegdić Vaso

    2013-01-01

    Full Text Available In the past two decades, companies have been mentioning achievement of sustainability in their activities as a target of companies, governments and non-profit organizations, although measuring the degree to which an organization conducts its activities in a sustainable manner, can be very difficult. Sustainable tourism development requires a process of planning and management that will unite the interests of various stakeholders in a sustainable and strategic way. It requires an understanding of the meaning of sustainable development and guiding values for promoting sustainable tourism. The paper points to the importance of cross-sector partnerships and the roles of different stakeholders in the planning of sustainable tourism projects. Special importance is given to the community of which a willingness to understand the impacts of tourism industry is expected, as well as various procedures of engagement in participatory planning, consensus building and conflict resolution among all stakeholders. The aim of this research is to find an optimal model of planning of sustainable tourism projects that would take into consideration the interests of all stakeholders and reflect the specificities imposed by the acceptance of the concept of sustainable development by all participants in the project.

  6. Advances in bacterial specific imaging

    Directory of Open Access Journals (Sweden)

    David Wareham

    2005-10-01

    Full Text Available Nuclear medicine is a powerful diagnostic technique able to detect inflammatory foci in human disease. A wide range of agents have been evaluated for their ability to distinguish lesions due to microbial infection from those due to sterile inflammation. Advances continue to be made on the use of radiolabelled antibiotics which as well as being highly specific in the diagnosis of infection may be useful in monitoring the treatment and course of disease. Here we provide an update on in-vitro and clinical studies with a number of established and novel radiopharmaceuticalsA medicina nuclear é uma técnica poderosa de diagnóstico capaz de detectar focos inflamatórios em doenças humanas. Uma ampla gama de agentes tem sido avaliada em sua capacidade de distinguir lesões, devidas a infecções microbianas daquelas causadas por inflamações estéreis. Avanços continuam sendo realizados no uso de antibióticos radiomarcados que, da mesma forma que têm sido usados no diagnóstico altamente específico de infecções, podem ser úteis na monitoração do tratamento e do curso da doença. Neste estudo, nós apresentamos uma atualização sobre estudos in vitro e clínicos, com alguns novos radiofármacos e outros já consagrados.

  7. Tumor specific cytotoxicity of glucosylceramide.

    Science.gov (United States)

    Oku, Hirosuke; Wongtangtintharn, Sawitree; Iwasaki, Hironori; Inafuku, Masashi; Shimatani, Masayuki; Toda, Takayoshi

    2007-11-01

    To develop a new taxon of anti-cancer agent with lower side effect, this study described a tumor selective cytotoxicity of glucosylceramide extracted from malt feed of beer brewing waste. Interpretation of (13)C- and (1)H-NMR spectra identified the chemical structure of major component of glucosylceramide as 1-O-beta-D: -glucopyranosyl-2(2'-hydroxyeicosanoylamino)-4,11-octadecadiene-1,3-diol. Selective cytotoxicity was studied with three pairs of normal and cancer cells: liver, skin and lung. The glucosylceramide selectively lowered the relative viability of cancer cells. Of the pairs, the selectivity was most pronounced with the liver cells, and, for this reason, further experiment was conducted with this pair of normal (CS-HC) and cancer cells (HepG2) to get more insight into the selective toxicity. The glucosylceramide significantly increased the cell population at G(2)/M phase in HepG2 cells, and also increased the numbers of apoptotic (sub-G(0)/G(1)) cells, but to much lesser extent compared with the increase in G(2)/M phase. Treatment of HepG2 cells with this agent selectively disrupted the mitochondrial membrane integrity without activation of caspase pathway to induce apoptosis. These findings suggested that the glucosylceramide specifically suppressed the growth of cancer cells by inhibiting cell renewal capacity rather than induction of apoptosis. The underlying mechanism for the selectivity remains to be answered in the forthcoming study.

  8. RNB-specific linac development

    CERN Document Server

    Ratzinger, U

    2002-01-01

    The acceleration of secondary, radioactive beams from primary beam driven ion sources provides many challenges when compared to the case of conventional ion beams. The clean separation of the specific ion species, the low beam current, the short life time of the ions as well as the needed energy variability have consequences on the layout of the accelerator. As also experiments with astrophysical relevance are performed, the energy range should go down below 1 MeV/u in some cases. Additionally, to investigate nuclear reactions around the Coulomb barrier, the beam energy should be flexible up to around 8 MeV/u. Short isotope life times below a few milliseconds do not allow for charge breeding. In these cases, low charged ions from 'simple' ion sources have to be accepted by the accelerator. Furthermore, stripping processes along the linac are excluded in case of low beam currents. Multicoincidence experiments profit a lot from cw operation of ion source and accelerators. During the last three decades, room tem...

  9. Are specific emotions narrated differently?

    Science.gov (United States)

    Habermas, Tilmann; Meier, Michaela; Mukhtar, Barbara

    2009-12-01

    Two studies test the assertion that anger, sadness, fear, pride, and happiness are typically narrated in different ways. Everyday events eliciting these 5 emotions were narrated by young women (Study 1) and 5- and 8-year-old girls (Study 2). Negative narratives were expected to engender more effort to process the event, be longer, more grammatically complex, more often have a complication section, and use more specific emotion labels than global evaluations. Narratives of Hogan's (2003) juncture emotions anger and fear were expected to focus more on action and to contain more core narrative sections of orientation, complication, and resolution than narratives of the outcome emotions sadness and happiness. Hypotheses were confirmed for adults except for syntactic complexity, whereas children showed only some of these differences. Hogan's theory that juncture emotions are restricted to the complication section was not confirmed. Finally, in adults, indirect speech was more frequent in anger narratives and internal monologue in fear narratives. It is concluded that different emotions should be studied in how they are narrated, and that narratives should be analyzed according to qualitatively different emotions.

  10. Specifications in early conceptual design work

    DEFF Research Database (Denmark)

    Hansen, Claus Thorp; Andreasen, Mogens Myrup

    2007-01-01

    specifications, which express attractive product goals, and has to synthesise the product idea. The authors of this paper see a challenge to enhance and improve our understanding of the nature of design specifications as a means to support the synthesis of a product idea. In this empirical study we analyse...... the structure and content of design specifications during early ideation activities, where initial design specifications are formulated and a product idea is synthesised. We have analysed specification documents of 19 teams of novice designers. Our analysis indicates that a productive product design...... specification in early conceptual design consists of few specification statements, which express value, important aspects of product context and articulate key functions. A product design specification, which contains a lot of specification statements about product properties, is not productive to support...

  11. Model Commissioning Plan and Guide Specifications

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-03-01

    The objectives of Model Commissioning Plan and Guide Specifications are to ensure that the design team applies commissioning concepts to the design and prepares commissioning specifications and a commission plan for inclusion in the bid construction documents.

  12. Ion specific effects on charged interfaces

    OpenAIRE

    Medda, Luca

    2013-01-01

    The physico-chemical phenomena occurring at charged interfaces are specifically affected by the type and the concentration of electrolytes. This has implications both in living and in inorganic systems. The discovery of the ‘ion specific effects’ dates back to Hofmeister (1888), who observed the specific effect of salts in promoting egg white proteins precipitation. Nowadays we are aware that ion specific effects are ubiquitous in all fields of science and technology where electrolytes play a...

  13. Normalized Specifications for Identifying Reusable Software

    OpenAIRE

    Luqi

    1987-01-01

    An approach to retrieving reusable software components by means of module specifications is described. The approach depends on normalizing specifications to reduce the variations in the representation of software concepts. The concept is illustrated in terms of both formal and informal approaches to component specifications.

  14. Programming Bacteriophages by Swapping Their Specificity Determinants.

    Science.gov (United States)

    Goren, Moran G; Yosef, Ido; Qimron, Udi

    2015-12-01

    Bacteriophages, bacteria's natural enemies, may serve as potent antibacterial agents. Their specificity for certain bacterial sub-species limits their effectiveness, but allows selective targeting of bacteria. Lu and colleagues present a platform for such targeting through alteration of bacteriophages' host specificity by swapping specificity domains in their host-recognition ligand.

  15. 31 CFR 515.318 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 515.318 Section 515.318 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... § 515.318 Specific license. A specific license is any license or authorization issued pursuant to...

  16. 31 CFR 535.318 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 535.318 Section 535.318 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... § 535.318 Specific license. A specific license is any license or authorization issued pursuant to...

  17. 31 CFR 585.308 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 585.308 Section 585.308 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... General Definitions § 585.308 Specific license. The term specific license means any license...

  18. 31 CFR 500.318 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 500.318 Section 500.318 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... § 500.318 Specific license. A specific license is any license or authorization issued pursuant to...

  19. 31 CFR 586.315 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 586.315 Section 586...) KOSOVO SANCTIONS REGULATIONS General Definitions § 586.315 Specific license. The term specific license means any license or authorization not set forth in this part but issued pursuant to this part....

  20. 31 CFR 536.313 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 536.313 Section 536.313 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... Definitions § 536.313 Specific license. The term specific license means any license or authorization not...

  1. 31 CFR 595.312 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 595.312 Section 595.312 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... § 595.312 Specific license. The term specific license means any license or authorization not set...

  2. 31 CFR 560.312 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 560.312 Section 560.312 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... § 560.312 Specific license. The term specific license means any license or authorization not set...

  3. 31 CFR 538.311 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 538.311 Section 538.311 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF... § 538.311 Specific license. The term specific license means any license or authorization not set...

  4. 50 CFR 648.260 - Specifications.

    Science.gov (United States)

    2010-10-01

    .... Specifications include the specification of OY, the setting of any target TACs, allocation of DAS, and/or... allocation based on target TAC. For purposes of determining the appropriate DAS allocation, any overage of... developing the management measures and specifications, the PDT shall review at least the following data,...

  5. The etiology of specific phobias : A review

    NARCIS (Netherlands)

    Merckelbach, H; de Jong, Peter; Muris, P; vandenHout, MA

    1996-01-01

    The present article summarizes theory and data about symptomatology, epidemiology, and etiology of specific phobias. Additionally, the cognitive mechanisms involved in specific Phobias are briefly discussed By and large, the general pattern behind the development of specific phobias can be summarize

  6. Genre-Specific Semantic Video Indexing

    NARCIS (Netherlands)

    Wu, J.; Worring, M.

    2010-01-01

    In many applications, we find large video collections from different genres where the user is often only interested in one or two specific video genres. So, when users are querying the system with a specific semantic concept, they are likely aiming a genre specific instantiation of this concept. Thu

  7. Map Specifications and Exchange of Geographical Information

    DEFF Research Database (Denmark)

    Frederiksen, Poul

    1999-01-01

    Specifications for Technical Maps 1993 – 99 are described giving an overview of the specification structure including the object description of the latest version: TK99.The technical map specifications are related to the standards for topographical maps - especially the TOP10DK standard. Common o...

  8. Checking MSC Specifications for Timing Inconsistency

    Institute of Scientific and Technical Information of China (English)

    LI Xuandong(李宣东); TAN Wenkai(谭文凯); ZHENG Guoliang(郑国梁)

    2002-01-01

    Message sequence chart (MSC) is a graphical and textual language for the description and specification of the interactions between system components. MSC specifica tions allow convenient expression of multiple scenarios, and offer an intuitive and visual way of describing design requirements. Like any other aspect of the specification and design process, MSCs are amenable to errors, and their analysis is important. In this paper, the verification problem of MSC specification for timing inconsistency is studied, which means that no execu tion scenario described by an MSC specification is timing consistent. An algorithm is developed to check MSC specifications for timing inconsistency.

  9. Inhibition of HIV-1 in Vitro by Combination of Vpr and Tat Specific Short Hairpin RNA via Lentiviral Vectors%慢病毒介导双shRNA靶向于HIV的抑制作用研究

    Institute of Scientific and Technical Information of China (English)

    郝彦哲; 滕智平; 杨怡姝; 孙晓娜; 马晶; 金孝华; 曾毅

    2013-01-01

    RNA干扰可以有效地抑制特定基因的表达,在HIV基因治疗中成为一种重要的方法.为了防止病毒逃逸株的出现,本研究分别构建了靶向于HIV-1调控蛋白基因vpr和tat的shRNA重组慢病毒载体以及含有这两种shRNA的双表达重组慢病毒载体,分别由U6和H1两种启动子启动,通过与靶向基因tat和vpr表达质粒在293T细胞中进行共转染,采用荧光定量PCR方法测定细胞内靶向基因的表达丰度,结果显示双表达shRNA的重组慢病毒载体对vpr和tat的表达抑制率分别可以达到89.20%和62.00%.与pNL4-3病毒包装质粒共转染,P24ELISA显示plvx-vpr-tat shRNA对病毒包装产量的抑制率可以达到99.05%,比单个shRNA的抑制率都要强,HIV-1 NL4-3体外对MT4细胞攻毒实验表明双shRNA重组慢病毒载体具有很强的抑制病毒复制的能力.%Gene therapy offers the promise of curing the HIV-infected patients. Specific, potent, and sustained short hairpin RNA (shRNA)-mediated gene silencing is crucial for the successful application of RNA interference technology to therapeutic interventions. To reduce the probability of viral escape mutants, in this study, we constructed lentiviral vector containing vpr and tat shRNA, respectively, furthermore the bispecific lentiviral vector harboring vpr and tat shRNA expression cassettes from U6 pro-motor and H1 promotor was cotransfected with recombinant plasmid expressing the vpr and tat gene. The result showed that the bispecific lentiviral vector plvx-vpr-tatshRNA could inhibit the vpr and tat effectively, with ratios of 89. 20% and 62. 00% respectively. When cotransfected with pNL4-3 in 293T cell, plvx-vpr-tatshRNA showed higher efficacy in down regulating the HIV NL4-3 packaging production than the plvx-vprshRNA or plvx-tatshRNA individually. MT4 cell clones transduced with recombinant lentiviral vectors were screened and challenged with HIV NL4-3. P24 ELISA test showed that MT4 transduced with the

  10. Towards a theory of product design specifications

    DEFF Research Database (Denmark)

    Hansen, Claus Thorp; Andreasen, Mogens Myrup

    2004-01-01

    The product design specification is an important element in a product development project because it defines the target to be met. One requirement on the product design specification is to articulate and communicate the aspects, which makes the product attractive seen from the users' viewpoint....... In the design methodology literature we find guidelines and methods to compile a product design specification. The contributions are based on the common underlying assumption that it is meaningful and the only feasible approach to interpret the result of a need analysis into a set of technical specifications......, which express the customers¿ need and perception of value. In this paper we will scrutinize this assumption. We outline the roles and tasks of the product design specification in order to identify the requirements on a theory of product design specifications and we identify existing theory elements...

  11. Peptide binding specificity of the chaperone calreticulin

    DEFF Research Database (Denmark)

    Sandhu, N.; Duus, K.; Jorgensen, C.S.;

    2007-01-01

    Calreticulin is a molecular chaperone with specificity for polypeptides and N-linked monoglucosylated glycans. In order to determine the specificity of polypeptide binding, the interaction of calreticulin with polypeptides was investigated using synthetic peptides of different length and composit......Calreticulin is a molecular chaperone with specificity for polypeptides and N-linked monoglucosylated glycans. In order to determine the specificity of polypeptide binding, the interaction of calreticulin with polypeptides was investigated using synthetic peptides of different length...... and composition. A large set of available synthetic peptides (n=127) was tested for binding to calreticulin and the results analysed by multivariate data analysis. The parameter that correlated best with binding was hydrophobicity while beta-turn potential disfavoured binding. Only hydrophobic peptides longer...... a peptide-binding specificity for hydrophobic sequences and delineate the fine specificity of calreticulin for hydrophobic amino acid residues....

  12. Specific fears and phobias. Epidemiology and classification.

    Science.gov (United States)

    Curtis, G C; Magee, W J; Eaton, W W; Wittchen, H U; Kessler, R C

    1998-09-01

    Data on eight specific fears representing DSM-III-R simple phobia were analysed to evaluate: (a) their prevalence and (b) the validity of subtypes of specific phobia defined by DSM-IV. A modified version of the Composite International Diagnostic Interview was administered to a probability sample of 8098 community respondents. Correlates of responses to questions concerning these fears were analysed. The most prevalent specific fears were of animals among women, and of heights among men. Slight evidence was found for specific phobia subtypes. Number of fears, independent of type, powerfully predicted impairment, comorbidity, illness course, demographic features, and family psychopathology. Number of specific fears may mark a general predisposition to psychopathology. More detailed information is needed to resolve the question of specific phobia subtypes.

  13. Using Abduction to Evolve Inconsistent Requirements Specification

    Directory of Open Access Journals (Sweden)

    Bashar Nuseibeh

    1999-05-01

    Full Text Available Requirements specifications are often inconsistent. Inconsistencies may arise because multiple conflicting requirements are embodied in these specifications, or because the specifications themselves are in a transient stage of evolutionary development. In this paper we argue that such inconsistencies, rather than being undesirable, are actually useful drivers for changing the requirements specifications in which they arise. We present a formal technique to reason about inconsistency handling changes. Our technique is an adaptation of logical abduction - adapted to generate changes that address some specification inconsistencies, while leaving others. We represent our specifications in quasi-classical (QC logic - an adaptation of classical logic that allows continued reasoning in the presence of inconsistency. The paper develops a sound algorithm for automating our abductive reasoning technique and presents illustrative examples drawn from a library system case study.

  14. Embedded systems specification and design languages

    CERN Document Server

    Villar, Eugenio

    2008-01-01

    This book is the latest contribution to the Chip Design Languages series and it consists of selected papers presented at the Forum on Specifications and Design Languages (FDL'07), in September 2007. The book represents the state-of-the-art in research and practice, and it identifies new research directions. It highlights the role of specification and modelling languages, and presents practical experiences with specification and modelling languages.

  15. Domain-Specific Ontology of Botany

    Institute of Scientific and Technical Information of China (English)

    Fang Gu; Cun-Gen Cao; Yue-Fei Sui; Wen Tian

    2004-01-01

    Domain-specific ontologies are greatly useful in knowledge acquisition, sharing and analysis. In this paper, botany-specific ontology for acquiring and analyzing botanical knowledge is presented. The ontology is represented in a set of well-defined categories, and each concept is viewed as an instance of certain category. The authors also introduce botany-specific axioms, an integral part of the ontology, for checking and reasoning with the acquired knowledge. Consistency, completeness and redundancy of the axioms are discussed.

  16. Standards and methods for software requirements specification

    OpenAIRE

    2014-01-01

    This thesis presents the comparison between three selected standards and methods for software requirements specifications. IEEE 830 specification and use cases represent older, while user stories represent newer generation of methods for specification writing. Each method is first explained in theory and then on a practical example. E-študent, a well-known application to the students of the Faculty of computer and information science, serves as our practical example. The application is onl...

  17. Certifiable Specification and Verification of C Programs

    Science.gov (United States)

    Lüth, Christoph; Walter, Dennis

    A novel approach to the specification and verification of C programs through an annotation language that is a mixture between JML and the language of Isabelle/HOL is proposed. This yields three benefits: specifications are concise and close to the underlying mathematical model; existing Isabelle theories can be reused; and the leap of faith from specification language to encoding in a logic is small. This is of particular relevance for software certification, and verification in application areas such as robotics.

  18. Construction and Ldentification of Silencing Vectors of Human and Mice Co-targeting ERβ-specific Gene%人鼠共打靶 ERβ特异性基因沉默载体的构建和鉴定

    Institute of Scientific and Technical Information of China (English)

    鲁世金; 孙长英

    2015-01-01

    Objective:To construct the silencing lentiviral vectors human and mice co-targeting ERβ-specific gene of human and mice.Methods:Retrieving the transcripts and sequence of the ERβgene of human and mice.Using the software SiRNA Target Finder to design human and mice co-targeting ERβ gene silencing siRNA and negative control,and compounding double stranded shRNA in shape of stem loop,being connected with linearized plasmid PLL3.7 by T4 DNA ligase to reconstruct as a recombinant plasmid (ERβ-shRNA-PLL3.7 and negative control plasmid ER-N-shRNA-PLL3.7), which were identified and transfected into human osteosarcoma cells MG63 and mouse osteoblastic cell line MC3T3-E1 cells. The testing ways of Realtime-PCR and western blot were used to test gene silencing effect of the recombinant plasmid.The SPSS1 6.0 software was used for statistical analysis,and the testing level was set at P <0.05.Results:It was confirmed by sequencing that the recombinant plasmid contained the shRNA sequence.After transfecting into mice osteoblastic cell line of MC3T3-E1 and human osteosarcoma cells of MG63,whether the ERβgene expression detected by Realtime-PCR,or Protein expression detected by the western blot,were decreased significantly compared with the control group or the negative control group,and there was a prominent difference,P <0.05.Conclusion:successfully constructed the ERβ-specific gene silencing lentiviral vectors of human and mice co-targeting effectively,and they were packaged into pseudoviral particles,which laid a foundation for the study of man gene function by using mouse as animal model or the development of gene-based drugs.%目的::构建人和鼠共打靶特异性 ERβ基因沉默慢病毒载体。方法:检索人和小鼠 ERβ基因转录本及其序列,应用 SiRNA Target Finder 软件设计人和小鼠共打靶 ERβ基因沉默 siRNA 和阴性对照siRNA,合成发夹状双链 shRNA,与线性化质粒 PLL3.7连接构建为重组质粒(ERβ-shRNA-PLL3.7

  19. Specifications in early conceptual design work

    DEFF Research Database (Denmark)

    Hansen, Claus Thorp; Andreasen, Mogens Myrup

    2007-01-01

    In early conceptual design the design team is working in an uncertain situation, where the understanding of a need is limited and not much is known about the solution space. In this situation the design team has to both analyse need and explore solution space. Thus, the team has to formulate design...... specification in early conceptual design consists of few specification statements, which express value, important aspects of product context and articulate key functions. A product design specification, which contains a lot of specification statements about product properties, is not productive to support...

  20. 77 FR 27814 - Model Safety Evaluation for Plant-Specific Adoption of Technical Specifications Task Force...

    Science.gov (United States)

    2012-05-11

    ... COMMISSION Model Safety Evaluation for Plant-Specific Adoption of Technical Specifications Task Force... availability. SUMMARY: The U.S. Nuclear Regulatory Commission (NRC) is announcing the availability of the model safety evaluation (SE) for plant-specific adoption of Technical Specifications (TSs) Task Force...

  1. 77 FR 27490 - Plant-Specific Adoption, Revision 4 of the Improved Standard Technical Specifications

    Science.gov (United States)

    2012-05-10

    ... Technical Specifications, NUREG-1430, ``Standard Technical Specifications, Babcock and Wilcox Plants....resource@nrc.gov . NUREG-1430, ``Standard Technical Specifications, Babcock and Wilcox Plants'' Revision 4... ML12100A177 ML12100A178 Specifications, Babcock and Wilcox Plants'' NUREG-1431, ``Standard...

  2. User Interface Technology for Formal Specification Development

    Science.gov (United States)

    Lowry, Michael; Philpot, Andrew; Pressburger, Thomas; Underwood, Ian; Lum, Henry, Jr. (Technical Monitor)

    1994-01-01

    Formal specification development and modification are an essential component of the knowledge-based software life cycle. User interface technology is needed to empower end-users to create their own formal specifications. This paper describes the advanced user interface for AMPHION1 a knowledge-based software engineering system that targets scientific subroutine libraries. AMPHION is a generic, domain-independent architecture that is specialized to an application domain through a declarative domain theory. Formal specification development and reuse is made accessible to end-users through an intuitive graphical interface that provides semantic guidance in creating diagrams denoting formal specifications in an application domain. The diagrams also serve to document the specifications. Automatic deductive program synthesis ensures that end-user specifications are correctly implemented. The tables that drive AMPHION's user interface are automatically compiled from a domain theory; portions of the interface can be customized by the end-user. The user interface facilitates formal specification development by hiding syntactic details, such as logical notation. It also turns some of the barriers for end-user specification development associated with strongly typed formal languages into active sources of guidance, without restricting advanced users. The interface is especially suited for specification modification. AMPHION has been applied to the domain of solar system kinematics through the development of a declarative domain theory. Testing over six months with planetary scientists indicates that AMPHION's interactive specification acquisition paradigm enables users to develop, modify, and reuse specifications at least an order of magnitude more rapidly than manual program development.

  3. Specific and social phobias in ICD-11

    NARCIS (Netherlands)

    Emmelkamp, P.M.G.

    2012-01-01

    The purpose of this article is to discuss the current status of research with respect to specific phobias and social phobia as defined in the ICD-10 and DSM-IV, and to provide op- tions for their classification in the ICD-11. Both ICD-10 and DSM-IV identify three main classes of phobias: specific

  4. 14 CFR 398.3 - Specific airports.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Specific airports. 398.3 Section 398.3... STATEMENTS GUIDELINES FOR INDIVIDUAL DETERMINATIONS OF BASIC ESSENTIAL AIR SERVICE § 398.3 Specific airports. (a) At an eligible place, essential air service may be specified as service to a particular...

  5. 28 CFR 0.131 - Specific functions.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Specific functions. 0.131 Section 0.131 Judicial Administration DEPARTMENT OF JUSTICE ORGANIZATION OF THE DEPARTMENT OF JUSTICE Bureau of Alcohol, Tobacco, Firearms, and Explosives § 0.131 Specific functions. The Director of the Bureau of...

  6. Steps in Modular Specifications for Concurrent Modules

    DEFF Research Database (Denmark)

    Da Rocha Pinto, Pedro; Dinsdale-Young, Thomas; Gardner, Philippa

    2015-01-01

    The specification of a concurrent program module is a difficult problem. The specifications must be strong enough to enable reasoning about the intended clients without reference to the underlying module implementation. We survey a range of verification techniques for specifying concurrent module...

  7. 28 CFR 0.101 - Specific functions.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Specific functions. 0.101 Section 0.101 Judicial Administration DEPARTMENT OF JUSTICE ORGANIZATION OF THE DEPARTMENT OF JUSTICE Drug Enforcement Administration § 0.101 Specific functions. The Administrator of the Drug Enforcement Administration shall...

  8. An Institution for Imperative RSL Specifications

    DEFF Research Database (Denmark)

    Haxthausen, Anne Elisabeth

    2014-01-01

    -oriented type definitions, declaration of state variables, axiomatic specification of values (including functions), and explicit function definitions. Functions may be imperative. The semantics of an RSLI specification is defined to be the loose semantics of a theory presentation consisting of a signature Σ...

  9. 21 CFR 107.100 - Nutrient specifications.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Nutrient specifications. 107.100 Section 107.100... FOR HUMAN CONSUMPTION INFANT FORMULA Nutrient Requirements § 107.100 Nutrient specifications. (a) An infant formula shall contain the following nutrients at a level not less than the minimum level...

  10. Specific heats of degenerate ideal gases

    OpenAIRE

    Caruso, Francisco; Oguri, Vitor; Silveira, Felipe

    2017-01-01

    From arguments based on Heisenberg's uncertainty principle and Pauli's exclusion principle, the molar specific heats of degenerate ideal gases at low temperatures are estimated, giving rise to values consistent with the Nerst-Planck Principle (third law of Thermodynamics). The Bose-Einstein condensation phenomenon based on the behavior of specific heat of massive and non-relativistic boson gases is also presented.

  11. Specific and social phobias in ICD-11

    NARCIS (Netherlands)

    Emmelkamp, P.M.G.

    2012-01-01

    The purpose of this article is to discuss the current status of research with respect to specific phobias and social phobia as defined in the ICD-10 and DSM-IV, and to provide op- tions for their classification in the ICD-11. Both ICD-10 and DSM-IV identify three main classes of phobias: specific ph

  12. SPECIFIC SKILLS AND SOCIAL COMPETENCE IN SCHIZOPHRENIA

    NARCIS (Netherlands)

    APPELO, MT; WOONINGS, FMJ; VANNIEUWENHUIZEN, CJ; EMMELKAMP, PMG; SLOOFF, CJ; LOUWERENS, JW

    1992-01-01

    Generalization of skills is a major problem in social skills training for schizophrenic patients. Assessment of skills is mostly not based on objective indices of specific skill deficits. The results of this study show that global competence of schizophrenics can be differentiated from specific comp

  13. Some Approaches for Integration of Specification Techniques

    DEFF Research Database (Denmark)

    Haxthausen, Anne Elisabeth

    2000-01-01

    It is often useful to apply several specification techniques within the same software development project. This raises the question how specification techniques can be integrated. In this presentation we give three different examples of how this can be done. In the first example, we summarise how...

  14. Plutonium Immobilization Can Loading Preliminary Specifications

    Energy Technology Data Exchange (ETDEWEB)

    Kriikku, E.

    1998-11-25

    This report discusses the Plutonium Immobilization can loading preliminary equipment specifications and includes a process block diagram, process description, equipment list, preliminary equipment specifications, plan and elevation sketches, and some commercial catalogs. This report identifies loading pucks into cans and backfilling cans with helium as the top priority can loading development areas.

  15. Specific phobia predicts psychopathology in young women

    NARCIS (Netherlands)

    Trumpf, J.; Margraf, J.; Vriends, N.; Meyer, A.H.; Becker, E.S.

    2010-01-01

    Although specific phobia is characterized by an early age at onset and by high rates of comorbidity, few studies have examined comorbid relationships prospectively. The present study investigated the association between specific phobia and the risk of a broad range of psychopathology among young wom

  16. Cell-Type-Specific Optogenetics in Monkeys.

    Science.gov (United States)

    Namboodiri, Vijay Mohan K; Stuber, Garret D

    2016-09-08

    The recent advent of technologies enabling cell-type-specific recording and manipulation of neuronal activity spurred tremendous progress in neuroscience. However, they have been largely limited to mice, which lack the richness in behavior of primates. Stauffer et al. now present a generalizable method for achieving cell-type specificity in monkeys.

  17. Determining the specific electric resistance of rock

    Energy Technology Data Exchange (ETDEWEB)

    Persad' ko, V.Ia.

    1982-01-01

    Data are presented on perfecting the method of laboratory determination of the specific electric resistance of a rock formation. The average error in determining the specific electric resistance of the core at various locations is no more than two percent with low resistance values (2-5 ohms).

  18. Understanding the substrate specificity of conventional calpains.

    Science.gov (United States)

    Sorimachi, Hiroyuki; Mamitsuka, Hiroshi; Ono, Yasuko

    2012-09-01

    Calpains are intracellular Ca(2+)-dependent Cys proteases that play important roles in a wide range of biological phenomena via the limited proteolysis of their substrates. Genetic defects in calpain genes cause lethality and/or functional deficits in many organisms, including humans. Despite their biological importance, the mechanisms underlying the action of calpains, particularly of their substrate specificities, remain largely unknown. Studies show that certain sequence preferences influence calpain substrate recognition, and some properties of amino acids have been related successfully to substrate specificity and to the calpains' 3D structure. The full spectrum of this substrate specificity, however, has not been clarified using standard sequence analysis algorithms, e.g., the position-specific scoring-matrix method. More advanced bioinformatics techniques were used recently to identify the substrate specificities of calpains and to develop a predictor for calpain cleavage sites, demonstrating the potential of combining empirical data acquisition and machine learning. This review discusses the calpains' substrate specificities, introducing the benefits of bioinformatics applications. In conclusion, machine learning has led to the development of useful predictors for calpain cleavage sites, although the accuracy of the predictions still needs improvement. Machine learning has also elucidated information about the properties of calpains' substrate specificities, including a preference for sequences over secondary structures and the existence of a substrate specificity difference between two similar conventional calpains, which has never been indicated biochemically.

  19. Fundamental Characteristics of Industrial Variant Specification Systems

    DEFF Research Database (Denmark)

    Hansen, Benjamin Loer; Hvam, Lars

    2004-01-01

    fundamental concepts related to this task, which are relevant to understand for academia and practitioners working with the subject. This is done through a description of variant specification tasks and typical aspects of system solutions. To support the description of variant specification tasks and systems...

  20. 14 CFR 23.1397 - Color specifications.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Color specifications. 23.1397 Section 23.1397 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT... Color specifications. Each position light color must have the applicable International Commission...

  1. 14 CFR 27.1397 - Color specifications.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Color specifications. 27.1397 Section 27... AIRWORTHINESS STANDARDS: NORMAL CATEGORY ROTORCRAFT Equipment Lights § 27.1397 Color specifications. Each position light color must have the applicable International Commission on Illumination...

  2. 14 CFR 25.1397 - Color specifications.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Color specifications. 25.1397 Section 25... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Equipment Lights § 25.1397 Color specifications. Each position light color must have the applicable International Commission on Illumination...

  3. 14 CFR 29.1397 - Color specifications.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Color specifications. 29.1397 Section 29... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Equipment Lights § 29.1397 Color specifications. Each position light color must have the applicable International Commission on Illumination...

  4. Introduction to the ISO specification language LOTOS

    NARCIS (Netherlands)

    Bolognesi, Tomasso; Brinksma, Ed

    1987-01-01

    LOTOS is a specification language that has been specifically developed for the formal description of the OSI (Open Systems Interconnection) architecture, although it is applicable to distributed, concurrent systems in general. In LOTOS a system is seen as a set of processes which interact and exchan

  5. Specific vulnerability of substantia nigra compacta neurons

    NARCIS (Netherlands)

    Smidt, M.P.; Giovanni, G.; Di Matteo, V.; Esposito, E.

    2009-01-01

    The specific loss of substantia nigra compacta (SNc) neurons in Parkinson’s disease (PD) has been the main driving force in initiating research efforts to unravel the apparent SNc-specific vulnerability. Initially, metabolic constraints due to high dopamine turnover have been the main focus in the a

  6. Task-specific dystonia: pathophysiology and management.

    Science.gov (United States)

    Sadnicka, Anna; Kassavetis, Panagiotis; Pareés, Isabel; Meppelink, Anne Marthe; Butler, Katherine; Edwards, Mark

    2016-09-01

    Task-specific dystonia is a form of isolated focal dystonia with the peculiarity of being displayed only during performance of a specific skilled motor task. This distinctive feature makes task-specific dystonia a particularly mysterious and fascinating neurological condition. In this review, we cover phenomenology and its increasingly broad-spectrum risk factors for the disease, critically review pathophysiological theories and evaluate current therapeutic options. We conclude by highlighting the unique features of task-specific dystonia within the wider concept of dystonia. We emphasise the central contribution of environmental risk factors, and propose a model by which these triggers may impact on the motor control of skilled movement. By viewing task-specific dystonia through this new lens which considers the disorder a modifiable disorder of motor control, we are optimistic that research will yield novel therapeutic avenues for this highly motivated group of patients.

  7. Fundamental Characteristics of Industrial Variant Specification Systems

    DEFF Research Database (Denmark)

    Hansen, Benjamin Loer; Hvam, Lars

    2004-01-01

    This paper focuses on the operational task of creating customised variants of industrial specifications (e.g. drawings, routings and bill-of-materials). Rooted in a lack of existing literature on the subject the paper describes the nature of variant specification systems. It introduces some funda...... examples. In general the paper discusses an important focus area within mass customization and build-to-order production: the nature of industrial variant specification systems.......This paper focuses on the operational task of creating customised variants of industrial specifications (e.g. drawings, routings and bill-of-materials). Rooted in a lack of existing literature on the subject the paper describes the nature of variant specification systems. It introduces some...

  8. Tissue Specific Promoters in Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    A. R. Rama

    2015-01-01

    Full Text Available Colorectal carcinoma is the third most prevalent cancer in the world. In the most advanced stages, the use of chemotherapy induces a poor response and is usually accompanied by other tissue damage. Significant progress based on suicide gene therapy has demonstrated that it may potentiate the classical cytotoxic effects in colorectal cancer. The inconvenience still rests with the targeting and the specificity efficiency. The main target of gene therapy is to achieve an effective vehicle to hand over therapeutic genes safely into specific cells. One possibility is the use of tumor-specific promoters overexpressed in cancers. They could induce a specific expression of therapeutic genes in a given tumor, increasing their localized activity. Several promoters have been assayed into direct suicide genes to cancer cells. This review discusses the current status of specific tumor-promoters and their great potential in colorectal carcinoma treatment.

  9. Tissue Specific Promoters in Colorectal Cancer

    Science.gov (United States)

    Rama, A. R.; Aguilera, A.; Melguizo, C.; Caba, O.; Prados, J.

    2015-01-01

    Colorectal carcinoma is the third most prevalent cancer in the world. In the most advanced stages, the use of chemotherapy induces a poor response and is usually accompanied by other tissue damage. Significant progress based on suicide gene therapy has demonstrated that it may potentiate the classical cytotoxic effects in colorectal cancer. The inconvenience still rests with the targeting and the specificity efficiency. The main target of gene therapy is to achieve an effective vehicle to hand over therapeutic genes safely into specific cells. One possibility is the use of tumor-specific promoters overexpressed in cancers. They could induce a specific expression of therapeutic genes in a given tumor, increasing their localized activity. Several promoters have been assayed into direct suicide genes to cancer cells. This review discusses the current status of specific tumor-promoters and their great potential in colorectal carcinoma treatment. PMID:26648599

  10. Mask specification guidelines in spacer patterning technology

    Science.gov (United States)

    Hashimoto, Kohji; Mukai, Hidefumi; Miyoshi, Seiro; Yamaguchi, Shinji; Mashita, Hiromitsu; Kobayashi, Yuuji; Kawano, Kenji; Hirano, Takashi

    2008-11-01

    We have studied both the mask CD specification and the mask defect specification for spacer patterning technology (SPT). SPT has the possibility of extending optical lithography to below 40nm half-pitch devices. Since SPT necessitates somewhat more complicated wafer process flow, the CD error and mask defect printability on wafers involve more process factors compared with conventional single-exposure process (SEP). This feature of SPT implies that it is very important to determine mask-related specifications for SPT in order to select high-end mask fabrication strategies; those are for mask writing tools, mask process development, materials, inspection tools, and so on. Our experimental studies reveal that both mask CD specification and mask defect specification are somehow relaxed from those in ITRS2007. This is most likely because SPT reduces mask CD error enhanced factor (MEF) and the reduction of line-width roughness (LWR).

  11. Specificity of transmembrane protein palmitoylation in yeast.

    Directory of Open Access Journals (Sweden)

    Ayelén González Montoro

    Full Text Available Many proteins are modified after their synthesis, by the addition of a lipid molecule to one or more cysteine residues, through a thioester bond. This modification is called S-acylation, and more commonly palmitoylation. This reaction is carried out by a family of enzymes, called palmitoyltransferases (PATs, characterized by the presence of a conserved 50- aminoacids domain called "Asp-His-His-Cys- Cysteine Rich Domain" (DHHC-CRD. There are 7 members of this family in the yeast Saccharomyces cerevisiae, and each of these proteins is thought to be responsible for the palmitoylation of a subset of substrates. Substrate specificity of PATs, however, is not yet fully understood. Several yeast PATs seem to have overlapping specificity, and it has been proposed that the machinery responsible for palmitoylating peripheral membrane proteins in mammalian cells, lacks specificity altogether.Here we investigate the specificity of transmembrane protein palmitoylation in S. cerevisiae, which is carried out predominantly by two PATs, Swf1 and Pfa4. We show that palmitoylation of transmembrane substrates requires dedicated PATs, since other yeast PATs are mostly unable to perform Swf1 or Pfa4 functions, even when overexpressed. Furthermore, we find that Swf1 is highly specific for its substrates, as it is unable to substitute for other PATs. To identify where Swf1 specificity lies, we carried out a bioinformatics survey to identify amino acids responsible for the determination of specificity or Specificity Determination Positions (SDPs and showed experimentally, that mutation of the two best SDP candidates, A145 and K148, results in complete and partial loss of function, respectively. These residues are located within the conserved catalytic DHHC domain suggesting that it could also be involved in the determination of specificity. Finally, we show that modifying the position of the cysteines in Tlg1, a Swf1 substrate, results in lack of palmitoylation, as

  12. Breckinridge Project, initial effort. Report III, Volume 2. Specifications

    Energy Technology Data Exchange (ETDEWEB)

    None

    1982-01-01

    Report III, Volume 2 contains those specifications numbered K through Y, as follows: Specifications for Compressors (K); Specifications for Piping (L); Specifications for Structures (M); Specifications for Insulation (N); Specifications for Electrical (P); Specifications for Concrete (Q); Specifications for Civil (S); Specifications for Welding (W); Specifications for Painting (X); and Specifications for Special (Y). The standard specifications of Bechtel Petroleum Incorporated have been amended as necessary to reflect the specific requirements of the Breckinridge Project and the more stringent specifications of Ashland Synthetic Fuels, Inc. These standard specifications are available for the Initial Effort (Phase Zero) work performed by all contractors and subcontractors.

  13. The HIV-Specific Advance Directive

    Science.gov (United States)

    Singer, Peter A; Thiel, Elaine C; Salit, Irving; Flanagan, William; Naylor, C. David

    1997-01-01

    OBJECTIVE To determine whether persons living with HIV find a disease-specific advance directive more acceptable than a generic directive. DESIGN Randomized clinical trial. SETTING HIV consumer organization and hospital-based HIV clinic. PARTICIPANTS Volunteer sample of persons with HIV. INTERVENTIONS The disease-specific HIV Living Will, the generic Centre for Bioethics Living Will, or both. MEASUREMENTS AND MAIN RESULTS Of 101 participants who received both advance directives, 78 (77.2%) preferred the disease-specific HIV Living Will and 23 (22.8%) preferred the generic Centre for Bioethics Living Will (p < .001). Most participants who preferred the HIV Living Will did so because it was more specific or relevant to their situation. CONCLUSIONS Persons living with HIV prefer a disease-specific to a generic advance directive. They should be offered a disease-specific advance directive. Our findings should also encourage investigators to develop and evaluate disease-specific advance directives in other clinical settings. PMID:9436891

  14. Validating Avionics Conceptual Architectures with Executable Specifications

    Directory of Open Access Journals (Sweden)

    Nils Fischer

    2012-08-01

    Full Text Available Current avionics systems specifications, developed after conceptual design, have a high degree of uncertainty. Since specifications are not sufficiently validated in the early development process and no executable specification exists at aircraft level, system designers cannot evaluate the impact of their design decisions at aircraft or aircraft application level. At the end of the development process of complex systems, e. g. aircraft, an average of about 65 per cent of all specifications have to be changed because they are incorrect, incomplete or too vaguely described. In this paper, a model-based design methodology together with a virtual test environment is described that makes complex high level system specifications executable and testable during the very early levels of system design. An aircraft communication system and its system context is developed to demonstrate the proposed early validation methodology. Executable specifications for early conceptual system architectures enable system designers to couple functions, architecture elements, resources and performance parameters, often called non-functional parameters. An integrated executable specification at Early Conceptual Architecture Level is developed and used to determine the impact of different system architecture decisions on system behavior and overall performance.

  15. Mining software specifications methodologies and applications

    CERN Document Server

    Lo, David

    2011-01-01

    An emerging topic in software engineering and data mining, specification mining tackles software maintenance and reliability issues that cost economies billions of dollars each year. The first unified reference on the subject, Mining Software Specifications: Methodologies and Applications describes recent approaches for mining specifications of software systems. Experts in the field illustrate how to apply state-of-the-art data mining and machine learning techniques to address software engineering concerns. In the first set of chapters, the book introduces a number of studies on mining finite

  16. Autoethnographic Mother-Writing: Advocating Radical Specificity

    Directory of Open Access Journals (Sweden)

    Patty Sotirin

    2010-01-01

    Full Text Available In considering the similarities between "momoirs"--popular memoirs written by mothers about motherhood experiences--and evocative autoethnographic mother-writing, I argue that differentiating these two forms of intimate observation and personal narrative requires a rethinking of autoethnographic practice. Specifically, I draw on the work of Gilles Deleuze to advocate for a radical specificity in autoethnographic writing. Thinking the autoethnographic narrative in terms of specificities and differences encourages us to think creatively about personal experiences and cultural relations beyond what is shared and communicable.

  17. PD-L1-specific T cells

    DEFF Research Database (Denmark)

    Ahmad, Shamaila Munir; Borch, Troels Holz; Hansen, Morten

    2016-01-01

    -specific T cells that recognize both PD-L1-expressing immune cells and malignant cells. Thus, PD-L1-specific T cells have the ability to modulate adaptive immune reactions by reacting to regulatory cells. Thus, utilization of PD-L1-derived T cell epitopes may represent an attractive vaccination strategy...... for targeting the tumor microenvironment and for boosting the clinical effects of additional anticancer immunotherapy. This review summarizes present information about PD-L1 as a T cell antigen, depicts the initial findings about the function of PD-L1-specific T cells in the adjustment of immune responses...

  18. Isothermal Gravitational Segregation: Algorithms and Specifications

    DEFF Research Database (Denmark)

    Halldórsson, Snorri; Stenby, Erling Halfdan

    2000-01-01

    New algorithms for calculating the isothermal equilibrium state of reservoir fluids under the influence of gravity are presented. Two types of specifications are considered: the specification of pressure and composition at a reference depth; and the specification of the total overall content...... of the reservoir, along with the reservoir geometry. It is shown how both types of calculations can be performed in an efficient and robust manner using volume-based thermodynamics. The new method makes it possible to evaluate the influence of reservoir geometry and gravity segregation on the hydrocarbon reserves...

  19. Specifications and programs for computer software validation

    Science.gov (United States)

    Browne, J. C.; Kleir, R.; Davis, T.; Henneman, M.; Haller, A.; Lasseter, G. L.

    1973-01-01

    Three software products developed during the study are reported and include: (1) FORTRAN Automatic Code Evaluation System, (2) the Specification Language System, and (3) the Array Index Validation System.

  20. FRENCH FOR SPECIFIC PURPOSES: THE GATEWAY TO ...

    African Journals Online (AJOL)

    JONATHAN

    2016-07-01

    Jul 1, 2016 ... acquire such skills in specific areas for accessing employment to live a meaningful life within the ... In concrete terms this policy considers the importance of ... neighbours, it is desirable for every Nigerian to speak French.

  1. 32 CFR 292.4 - Specific policy.

    Science.gov (United States)

    2010-07-01

    ... INFORMATION ACT PROGRAM DEFENSE INTELLIGENCE AGENCY (DIA) FREEDOM OF INFORMATION ACT § 292.4 Specific policy... characteristics, made or received by the DIA in connection with the transaction of public business and in the DIA...

  2. Female-specific DNA sequences in geese.

    Science.gov (United States)

    Huang, M C; Lin, W C; Horng, Y M; Rouvier, R; Huang, C W

    2003-07-01

    1. The OPAE random primers (Operon Technologies, Inc., CA) were used for random amplified polymorphic DNA (RAPD) fingerprinting in Chinese, White Roman and Landaise geese. One of these primers, OPAE-06, produced a 938-bp sex-specific fragment in all females and in no males of Chinese geese only. 2. A novel female-specific DNA sequence in Chinese goose was cloned and sequenced. Two primers, CGSex-F and CGSex-R, were designed in order to amplify a 912-bp sex-specific polymerase chain reaction (PCR) fragment on genomic DNA from female geese. 3. It was shown that a simple and effective PCR-based sexing technique could be used in the three goose breeds studied. 4. Nucleotide sequencing of the sex-specific fragments in White Roman and Landaise geese was performed and sequence differences were observed among these three breeds.

  3. Thermodynamic stability contributes to immunoglobulin specificity.

    Science.gov (United States)

    Dimitrov, Jordan D; Kaveri, Srinivas V; Lacroix-Desmazes, Sébastien

    2014-05-01

    Antigen-binding specificity of immunoglobulins is important for their function in immune defense. However, immune repertoires contain a considerable fraction of immunoglobulins with promiscuous binding behavior, the physicochemical basis of which is not well understood. Evolution of immunoglobulin specificity occurs through iterative processes of mutation and selection, referred to as affinity maturation. Recent studies reveal that some somatic mutations could compromise the thermodynamic stability of the variable regions of immunoglobulins. By integrating this observation with the wealth of data on the evolution of novel enzyme activities, we propose that antibody specificity is linked to the thermodynamic stability of the antigen-binding regions, which provides a quantitative distinction between highly specific and promiscuous antibodies.

  4. Specification Editing and Discovery Assistant Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The project will prototype a specification editing and discovery tool (SPEEDY) for C/C++ that will assist software developers with modular formal verification tasks...

  5. Modal Specifications for Probabilistic Timed Systems

    Directory of Open Access Journals (Sweden)

    Tingting Han

    2013-06-01

    Full Text Available Modal automata are a classic formal model for component-based systems that comes equipped with a rich specification theory supporting abstraction, refinement and compositional reasoning. In recent years, quantitative variants of modal automata were introduced for specifying and reasoning about component-based designs for embedded and mobile systems. These respectively generalize modal specification theories for timed and probabilistic systems. In this paper, we define a modal specification language for combined probabilistic timed systems, called abstract probabilistic timed automata, which generalizes existing formalisms. We introduce appropriate syntactic and semantic refinement notions and discuss consistency of our specification language, also with respect to time-divergence. We identify a subclass of our models for which we define the fundamental operations for abstraction, conjunction and parallel composition, and show several compositionality results.

  6. Site-specificity in small architectural interventions

    DEFF Research Database (Denmark)

    Marie Bruun Jespersen, Line

    2016-01-01

    This paper is investigating the concepts of site-specificity, place and experience as they have been interpreted in a number of projects with at strong emphasis on the characteristics of the site, in the form of small architectural or artistic interventions. The idea of site-specificity is re......-emerging in contemporary architectural discourse in a series of projects, in e.g. the Norwegian project ”National Tourist Roads”. The small architectural interventions consequently work with enhancing or framing the qualities of site and aim at being a mediating element between man and nature. In this paper two main...... specificity in the small architectural interventions. The paper concludes that the small architectural interventions operate with a narrow concept of site, and that the large variety of artistic strategies and types of site-specificity found in contemporary art could serve as inspiration to open up...

  7. Standard specification for silvered flat glass mirror

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2008-01-01

    1.1 This specification covers the requirements for silvered flat glass mirrors of rectangular shape supplied as cut sizes, stock sheets or as lehr ends and to which no further processing (such as edgework or other fabrication) has been done. 1.2 This specification covers the quality requirements of silvered annealed monolithic clear and tinted flat glass mirrors up to 6 mm (¼ in.) thick. The mirrors are intended to be used indoors for mirror glazing, for components of decorative accessories or for similar uses. 1.3 This specification does not address safety glazing materials nor requirements for mirror applications. Consult model building codes and other applicable standards for safety glazing applications. 1.4 Mirrors covered in this specification are not intended for use in environments where high humidity or airborne corrosion promoters, or both, are consistently present (such as swimming pool areas, ocean-going vessels, chemical laboratories and other corrosive environments). 1.5 The dimensional val...

  8. Specific detection of proteins using Nanomechanical resonators

    DEFF Research Database (Denmark)

    Fischer, Lee MacKenzie; Wright, V.A.; Guthy, C.;

    2008-01-01

    of probes onto their surfaces in order to enable the specificity of the detection. Such nanoresonator-based specific detection of proteins is here reported using streptavidin as target system, and immobilized biotin as probe. Nanomechanical resonators resistant to stiction were first realized from silicon...... carbonitride using a novel fabrication method. Vapor-phase deposition of mercaptopropyl trimethoxysilane was performed, and an added mass of 2.22 +/- 0.07 fg/mu m(2) was measured. This linker molecule was used to attach biotin onto the devices, enabling the specific detection of streptavidin. A mass of 3.6 fg....../mu m(2) was attributed to the added streptavidin, corresponding to one molecule per 27 nm(2). The specificity of this recognition was confirmed by exposing the devices to a solution of streptavidin that was already saturated with biotin. An additional negative control was also performed by also...

  9. SPECIFICS OF DRUG PHARMACOKINETICS IN PREGNANCY

    Directory of Open Access Journals (Sweden)

    A.V. Ostrovskaya

    2010-01-01

    Full Text Available Applying clinical pharmacology guidelines to the mother-fetus system makes it possible to better understand the specifics of pharmacokinetics during pregnancy. The article illustrates the factors that drive these specific features related to both changes in the boy of a future mother and the presence of an extra fetal-placental blood circulation, especially the placenta and developing fetus. These factors influence the results of drug-based treatment during the entire gestational period. Thanks to a rapid development of molecular technologies in the last decade, modern medicine has good prospects of answering questions about individual specifics of pharmacokinetics and metabolism of drugs, increased teratogenic risk driven by specifics of genotypes of the mother and fetus. Key words: pharmacokinetics, drugs, pregnancy, placenta, mother-fetus system. (Pediatric Pharmacology. – 2010; 7(5:44-47

  10. Frameworks of competence: common or specific?

    NARCIS (Netherlands)

    Grant, Simon

    2006-01-01

    Please, cite this publication as: Grant, S. (2006). Frameworks of competence: common or specific?. Proceedings of International Workshop in Learning Networks for Lifelong Competence Development, TENCompetence Conference. September 12th, Sofia, Bulgaria: TENCompetence. Retrieved June 30th, 2006, from

  11. Improved material specifications for unsealed roads

    CSIR Research Space (South Africa)

    Paige-Green, P

    2007-01-01

    Full Text Available into account so that the specifications can be effectively applied. A revised material selection guide and performance prediction chart have been developed for use when the materials are tested using the British Standard test methods....

  12. Cdc42-mediated tubulogenesis controls cell specification

    DEFF Research Database (Denmark)

    Kesavan, Gokul; Sand, Fredrik Wolfhagen; Greiner, Thomas Uwe

    2009-01-01

    Understanding how cells polarize and coordinate tubulogenesis during organ formation is a central question in biology. Tubulogenesis often coincides with cell-lineage specification during organ development. Hence, an elementary question is whether these two processes are independently controlled......, or whether proper cell specification depends on formation of tubes. To address these fundamental questions, we have studied the functional role of Cdc42 in pancreatic tubulogenesis. We present evidence that Cdc42 is essential for tube formation, specifically for initiating microlumen formation and later...... for maintaining apical cell polarity. Finally, we show that Cdc42 controls cell specification non-cell-autonomously by providing the correct microenvironment for proper control of cell-fate choices of multipotent progenitors. For a video summary of this article, see the PaperFlick file with the Supplemental Data...

  13. Cockpit management and Specific Behavioral Objectives (SBOs)

    Science.gov (United States)

    Mudge, R. W.

    1987-01-01

    One of the primary tools used to accomplish the task of effective training is the specific behavioral objective (SBO). An SBO is simply a statement which specifically identifies a small segment of the final behavior sought, and a little more. The key word is specific. The company pinpoints exactly what it is it wants the pilot to do after completing training, and what it should evaluate from the point of view of both the program and the pilot. It tells the junior crewmember exactly, specifically, what he should monitor and support insofar as the management function is concerned. It gives greater meaning to the term second in command. And finally, it tells the supervisory pilot exactly what he should observe, evaluate, and instruct, insofar as the management function is concerned.

  14. Physician Fee Schedule Carrier Specific Files

    Data.gov (United States)

    U.S. Department of Health & Human Services — The Centers for Medicare and Medicaid Services (CMS) has condensed all 56 Physician Fee Schedule (PFS) carrier specific pricing files into one zip file. It is...

  15. Gender-Specific Health Challenges Facing Women

    Science.gov (United States)

    ... with twitter share with linkedin Gender-Specific Health Challenges Facing Women Global Research Global Research NIAID's Role ... Career Stage Postdocs' Guide to Gaining Independence Small Business Programs Compare NIAID’s Small Business Programs High-Priority ...

  16. Specific heat and thermal conductivity of nanomaterials

    Science.gov (United States)

    Bhatt, Sandhya; Kumar, Raghuvesh; Kumar, Munish

    2017-01-01

    A model is proposed to study the size and shape effects on specific heat and thermal conductivity of nanomaterials. The formulation developed for specific heat is based on the basic concept of cohesive energy and melting temperature. The specific heat of Ag and Au nanoparticles is reported and the effect of size and shape has been studied. We observed that specific heat increases with the reduction of particle size having maximum shape effect for spherical nanoparticle. To provide a more critical test, we extended our model to study the thermal conductivity and used it for the study of Si, diamond, Cu, Ni, Ar, ZrO2, BaTiO3 and SrTiO3 nanomaterials. A significant reduction is found in the thermal conductivity for nanomaterials by decreasing the size. The model predictions are consistent with the available experimental and simulation results. This demonstrates the suitability of the model proposed in this paper.

  17. Interior Design's Role in Educational Specifications.

    Science.gov (United States)

    Keller, Mary Ann

    1986-01-01

    An experienced interior designer, equipped with a well-written educational specification, will specify the best materials and coordinate colors, materials, and furnishings in line with the district's budget. (MLF)

  18. Surface cleanliness of fluid systems, specification for

    Science.gov (United States)

    1995-01-01

    This specification establishes surface cleanliness levels, test methods, cleaning and packaging requirements, and protection and inspection procedures for determining surface cleanliness. These surfaces pertain to aerospace parts, components, assemblies, subsystems, and systems in contact with any fluid medium.

  19. Testing data types implementations from algebraic specifications

    CERN Document Server

    Gaudel, Marie-Claude

    2008-01-01

    Algebraic specifications of data types provide a natural basis for testing data types implementations. In this framework, the conformance relation is based on the satisfaction of axioms. This makes it possible to formally state the fundamental concepts of testing: exhaustive test set, testability hypotheses, oracle. Various criteria for selecting finite test sets have been proposed. They depend on the form of the axioms, and on the possibilities of observation of the implementation under test. This last point is related to the well-known oracle problem. As the main interest of algebraic specifications is data type abstraction, testing a concrete implementation raises the issue of the gap between the abstract description and the concrete representation. The observational semantics of algebraic specifications bring solutions on the basis of the so-called observable contexts. After a description of testing methods based on algebraic specifications, the chapter gives a brief presentation of some tools and case st...

  20. TELNET under Single-Connection TCP Specification

    Science.gov (United States)

    1976-02-02

    TTYOUTPUTINTERRUPTSERVICE, COMPOSEHESSAGE, AUAKENPRINTER, TYPENEXT. TYPE. nini -Telnet Implementation February 2, 197B Each list of routines is... nini -Telnet Implementation February 2. 1976 References 1 V. Cerf, Y. Dalai, C. Sunshine, "Specification of Internet

  1. Specification For ST-5 Li Ion Battery

    Science.gov (United States)

    Castell, Karen D.; Day, John H. (Technical Monitor)

    2000-01-01

    This Specification defines the general requirements for rechargeable Space Flight batteries intended for use in the ST-5 program. The battery chemistry chosen for this mission is lithium ion (Li-Ion).

  2. SPECIFIC KNOWLEDGE FOR MANAGING ECOTOURISM DESTINATIONS

    OpenAIRE

    Smolčić Jurdana, Dora

    2009-01-01

    Management of ecotourism destinations is very demanding because of the specific and interdisciplinary knowledge that is necessary. Great knowledge, which consists of knowledge from human and natural sciences at the same time, emphasizes the need for permanent education, formal and informal of ecotourism destination managers. In should be generally stated that countries which have a great areas and resources for ecotourism development should create a specific program for education about the...

  3. Specification and Construction of Control Flow Semantics

    OpenAIRE

    Smelik, R.; Rensink, A.; Kastenberg, H.; Grundy, J.; Howse, J.

    2006-01-01

    In this paper we propose a visual language CFSL for specifying control flow semantics of programming languages. We also present a translation from CFSL to graph production systems (GPS) for flow graph construction; that is, any CFSL specification, say for a language L, gives rise to a GPS that constructs from any L-program (represented as an abstract syntax graph) the corresponding flow graph. The specification language is rich enough to capture complex language constructs, including all of J...

  4. Patient-Specific Modeling in Tomorrow's Medicine

    CERN Document Server

    2012-01-01

    This book reviews the frontier of research and clinical applications of Patient Specific Modeling, and provides a state-of-the-art update as well as perspectives on future directions in this exciting field. The book is useful for medical physicists, biomedical engineers and other engineers who are interested in the science and technology aspects of Patient Specific Modeling, as well as for radiologists and other medical specialists who wish to be updated about the state of implementation.

  5. Measuring Specific Heats at High Temperatures

    Science.gov (United States)

    Vandersande, Jan W.; Zoltan, Andrew; Wood, Charles

    1987-01-01

    Flash apparatus for measuring thermal diffusivities at temperatures from 300 to 1,000 degrees C modified; measures specific heats of samples to accuracy of 4 to 5 percent. Specific heat and thermal diffusivity of sample measured. Xenon flash emits pulse of radiation, absorbed by sputtered graphite coating on sample. Sample temperature measured with thermocouple, and temperature rise due to pulse measured by InSb detector.

  6. TRIO specification of a steam boiler controller

    Energy Technology Data Exchange (ETDEWEB)

    Gargantini, A. [Politecnico di Milano (Italy). Dipt. di Electronica e Informazione; Morzenti, A. [Politecnico di Milano (Italy). Dipt. di Electronica e Informazione

    1996-12-31

    We specify a controller for a steam boiler starting from an informal descriptions of its requirements. The specification is formalized in the temporal logic TRIO and its object-oriented extension TRIO+. To obtain a maximum of abstraction and reuse we make the specification parametric with respect to all equipment and hardware features, and we avoid to impose any particular strategy in the management of the available resources and in the control of the critical physical quantities. (orig.)

  7. Simple Equational Specifications of Rational Arithmetic

    Directory of Open Access Journals (Sweden)

    Lawrence S. Moss

    2001-12-01

    Full Text Available We exhibit an initial specification of the rational numbers equipped with addition, subtraction, multiplication, greatest integer function, and absolute value. Our specification uses only the sort of rational numbers. It uses one hidden function; that function is unary. But it does not use an error constant, or extra (hidden sorts, or conditional equations. All of our work is elementary and self-contained.

  8. Reflections on the specificity of synaptic connections.

    Science.gov (United States)

    White, Edward L

    2007-10-01

    The principal focus of this treatise is the specificity of synaptic connectivity in the mammalian central nervous system. The occurrence of stereotypical patterns of connection at the macro level (e.g., the general consistency with which axonal pathways impinge on and originate within specific cortical areas and layers) implies that the cerebral cortex is a highly ordered structure. Order is seen also at the more micro level of synaptic connectivity, for instance, in the contrasting synaptic patterns of spiny vs. non-spiny neurons. Quantitative electron microscopic studies of synapses between identified neurons and correlative anatomical/electrophysiological investigations indicate that the high degree of order characterizing many aspects of cortical organization is mirrored by an equally ordered arrangement of synaptic connections between specific types of neurons. The recognition of recurring synaptic patterns has generated increased support for the notion of synaptic specificity as opposed to randomness, and we have begun now to understand the role of specificity in cortical function. At the core of cortical processing lie myriad possibilities for computation provided by the wealth of synaptic connections involving each neuron. Specificity, by limiting possibilities for connection, imposes an order on synaptic interactions even as processes of dynamic selection or synaptic remodeling ensure the constant formation and dissolution of cortical circuits. Collectively, these operations make maximal use of the richness of cortical synaptic connections to produce a highly flexible system, irrespective of the degree of hard-wiring, mutability, randomness or specificity that obtains for cortical wiring at any particular time. A brief, historical account of developments leading to our current understanding of cortical synaptic organization will precede the presentation of evidence for synaptic specificity.

  9. Specific phobia predicts psychopathology in young women

    OpenAIRE

    Trumpf, Julia; Margraf, Jürgen; Vriends, Noortje; Meyer, Andrea Hans; Becker, Eni S

    2009-01-01

    Background Although specific phobia is characterized by an early age at onset and by high rates of comorbidity, few studies have examined comorbid relationships prospectively. Objectives The present study investigated the association between specific phobia and the risk of a broad range of psychopathology among young women in the community. Method Data came from the Dresden Predictor Study in which 1,538 German women (18–25 years) completed a diagnostic interview at two time points. Results W...

  10. Language translation, doman specific languages and ANTLR

    Science.gov (United States)

    Craymer, Loring; Parr, Terence

    2002-01-01

    We will discuss the features of ANTLR that make it an attractive tool for rapid developement of domain specific language translators and present some practical examples of its use: extraction of information from the Cassini Command Language specification, the processing of structured binary data, and IVL--an English-like language for generating VRML scene graph, which is used in configuring the jGuru.com server.

  11. Military Specification Updates & Deicing Working Group Initiatives

    Science.gov (United States)

    2011-08-18

    Air Force Petroleum Agency I n t e g r i t y - S e r v i c e - E x c e l l e n c e Military Specification Updates & Deicing Working Group...00-2011 4. TITLE AND SUBTITLE Military Specification Updates & Deicing Working Group Initiatives 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c...Corrosion Runway Deicing Fluid Partnership with Battelle Memorial Institute (lead agency), AFRL/ ASC, Army (CRREL), Navy (NAVAIR), Octagon, AMIL

  12. Simple Benchmark Specifications for Space Radiation Protection

    Science.gov (United States)

    Singleterry, Robert C. Jr.; Aghara, Sukesh K.

    2013-01-01

    This report defines space radiation benchmark specifications. This specification starts with simple, monoenergetic, mono-directional particles on slabs and progresses to human models in spacecraft. This report specifies the models and sources needed to what the team performing the benchmark needs to produce in a report. Also included are brief descriptions of how OLTARIS, the NASA Langley website for space radiation analysis, performs its analysis.

  13. Structure and specificity of lamprey monoclonal antibodies

    OpenAIRE

    Herrin, Brantley R.; Alder, Matthew N; Roux, Kenneth H.; Sina, Christina; Ehrhardt, Götz R. A.; Boydston, Jeremy A.; Turnbough, Charles L.; Cooper, Max D.

    2008-01-01

    Adaptive immunity in jawless vertebrates (lamprey and hagfish) is mediated by lymphocytes that undergo combinatorial assembly of leucine-rich repeat (LRR) gene segments to create a diverse repertoire of variable lymphocyte receptor (VLR) genes. Immunization with particulate antigens induces VLR-B-bearing lymphocytes to secrete antigen-specific VLR-B antibodies. Here, we describe the production of recombinant VLR-B antibodies specific for BclA, a major coat protein of Bacillus anthracis spores...

  14. Tissue-specific and substrate-specific mitochondrial bioenergetics in feline cardiac and skeletal muscles

    DEFF Research Database (Denmark)

    Christiansen, Liselotte Bruun; Dela, Flemming; Koch, Jørgen;

    2015-01-01

    No studies have investigated the mitochondrial function in permeabilized muscle fiber from cats. The aim of this study was to investigate tissue-specific and substrate-specific characteristics of mitochondrial oxidative phosphorylation (OXPHOS) capacity in feline permeabilized oxidative muscle...

  15. Germ cell specification and regeneration in planarians.

    Science.gov (United States)

    Newmark, P A; Wang, Y; Chong, T

    2008-01-01

    In metazoans, two apparently distinct mechanisms specify germ cell fate: Determinate specification (observed in animals including Drosophila, Caenorhabditis elegans, zebra fish, and Xenopus) uses cytoplasmic factors localized to specific regions of the egg, whereas epigenetic specification (observed in many basal metazoans, urodeles, and mammals) involves inductive interactions between cells. Much of our understanding of germ cell specification has emerged from studies of model organisms displaying determinate specification. In contrast, our understanding of epigenetic/inductive specification is less advanced and would benefit from studies of additional organisms. Freshwater planarians--widely known for their remarkable powers of regeneration--are well suited for studying the mechanisms by which germ cells can be induced. Classic experiments showed that planarians can regenerate germ cells from body fragments entirely lacking reproductive structures, suggesting that planarian germ cells could be specified by inductive signals. Furthermore, the availability of the genome sequence of the planarian Schmidtea mediterranea, coupled with the animal's susceptibility to systemic RNA interference (RNAi), facilitates functional genomic analyses of germ cell development and regeneration. Here, we describe recent progress in studies of planarian germ cells and frame some of the critical unresolved questions for future work.

  16. Standardized Technical Specifications for Westinghouse PWRs

    Energy Technology Data Exchange (ETDEWEB)

    1978-06-15

    This Standard Technical Specification (STS) has been structured for the broadest possible use on Westinghouse plants currently being reviewed for an Operating License. Accordingly, the document contains specifications applicable to plants (1) with either 3 or 4 loops and (2) with and without loop stop valves. In addition, four separate and discrete containment specification sections are provided for each of the following containment types: Atmospheric, Ice Condenser, Sub-Atmospheric, and Dual. Optional specifications are provided for those features and systems which may be included in individual plant designs but are not generic in their scope of application. Alternate specifications are provided in a limited number of cases to cover situations where alternate specification requirements are necessary on a generic basis because of design differences. The format of the STS addresses the categories required by 10 CFR 50 and consists of six sections covering the areas of: Definitions, Safety Limits and Limiting Safety System Settings, Limiting Conditions for Operation, Surveillance Requirements, Design Features, and Administrative Controls.

  17. Age-specific reference ranges of serum prostate-specific antigen in Iranian men

    Directory of Open Access Journals (Sweden)

    Gholamreza Pourmand

    2015-08-01

    Conclusion: Findings of the present study showed that PSA levels are correlated with age. It was also revealed that the PSA age-specific reference range obtained in this study is different from other races and is specific to Iranian men. Therefore, age-specific reference ranges of PSA obtained in the present study can increase PSA test sensitivity and specificity by reducing unnecessary diagnostic procedures and early detection of prostate cancer in Iranian men.

  18. New primers for methylation-specific polymerase chain reaction enhance specificity of detecting STAT1 methylation

    Directory of Open Access Journals (Sweden)

    Ming-Cheng Chang

    2012-03-01

    Conclusion: Specific primers for methylation-specific PCR are mandatory for the accurate detection of STAT1 gene methylation. Besides, specific primers can generate correct interpretation of STAT1 gene methylation, and its correlation with the clinicopathological characteristics and outcome of cancer patients.

  19. Ultrasensitive Responses and Specificity in Cell Signaling

    Directory of Open Access Journals (Sweden)

    Haney Seth

    2010-08-01

    Full Text Available Abstract Background Interconnected cell signaling pathways are able to efficiently and accurately transmit a multitude of different signals, despite an inherent potential for undesirable levels of cross-talk. To ensure that an appropriate response is produced, biological systems have evolved network-level mechanisms that insulate pathways from crosstalk and prevent 'leaking' or 'spillover' between pathways. Many signaling pathways have been shown to respond in an ultrasensitive (switch-like fashion to graded input, and this behavior may influence specificity. The relationship of ultrasensitivity to signaling specificity has not been extensively explored. Results We studied the behavior of simple mathematical models of signaling networks composed of two interconnected pathways that share an intermediate component, asking if the two pathways in the network could exhibit both output specificity (preferentially activate their own output and input fidelity (preferentially respond to their own input. Previous results with weakly-activated pathways indicated that neither mutual specificity nor mutual fidelity were obtainable in the absence of an insulating mechanism, such as cross-pathway inhibition, combinatorial signaling or scaffolding/compartmentalization. Here we found that mutual specificity is obtainable for hyperbolic or ultrasensitive pathways, even in the absence of an insulating mechanism. However, mutual fidelity is impossible at steady-state, even if pathways are hyperbolic or ultrasensitive. Nevertheless, ultrasensitivity does provide advantages in attaining specificity and fidelity to networks that contain an insulating mechanism. For networks featuring cross-pathway inhibition or combinatorial signaling, ultrasensitive activation can increase specificity in a limited way, and can only be utilized by one of the two pathways. In contrast, for networks featuring scaffolding/compartmentalization, ultrasensitive activation of both pathways

  20. Face recognition: a model specific ability

    Directory of Open Access Journals (Sweden)

    Jeremy B Wilmer

    2014-10-01

    Full Text Available In our everyday lives, we view it as a matter of course that different people are good at different things. It can be surprising, in this context, to learn that most of what is known about cognitive ability variation across individuals concerns the broadest of all cognitive abilities, often labeled g. In contrast, our knowledge of specific abilities, those that correlate little with g, is severely constrained. Here, we draw upon our experience investigating an exceptionally specific ability, face recognition, to make the case that many specific abilities could easily have been missed. In making this case, we derive key insights from earlier false starts in the measurement of face recognition’s variation across individuals, and we highlight the convergence of factors that enabled the recent discovery that this variation is specific. We propose that the case of face recognition ability illustrates a set of tools and perspectives that could accelerate fruitful work on specific cognitive abilities. By revealing relatively independent dimensions of human ability, such work would enhance our capacity to understand the uniqueness of individual minds.

  1. Formal specification level concepts, methods, and algorithms

    CERN Document Server

    Soeken, Mathias

    2015-01-01

    This book introduces a new level of abstraction that closes the gap between the textual specification of embedded systems and the executable model at the Electronic System Level (ESL). Readers will be enabled to operate at this new, Formal Specification Level (FSL), using models which not only allow significant verification tasks in this early stage of the design flow, but also can be extracted semi-automatically from the textual specification in an interactive manner.  The authors explain how to use these verification tasks to check conceptual properties, e.g. whether requirements are in conflict, as well as dynamic behavior, in terms of execution traces. • Serves as a single-source reference to a new level of abstraction for embedded systems, known as the Formal Specification Level (FSL); • Provides a variety of use cases which can be adapted to readers’ specific design flows; • Includes a comprehensive illustration of Natural Language Processing (NLP) techniques, along with examples of how to i...

  2. Chromosome-specific DNA Repeat Probes

    Energy Technology Data Exchange (ETDEWEB)

    Baumgartner, Adolf; Weier, Jingly Fung; Weier, Heinz-Ulrich G.

    2006-03-16

    In research as well as in clinical applications, fluorescence in situ hybridization (FISH) has gained increasing popularity as a highly sensitive technique to study cytogenetic changes. Today, hundreds of commercially available DNA probes serve the basic needs of the biomedical research community. Widespread applications, however, are often limited by the lack of appropriately labeled, specific nucleic acid probes. We describe two approaches for an expeditious preparation of chromosome-specific DNAs and the subsequent probe labeling with reporter molecules of choice. The described techniques allow the preparation of highly specific DNA repeat probes suitable for enumeration of chromosomes in interphase cell nuclei or tissue sections. In addition, there is no need for chromosome enrichment by flow cytometry and sorting or molecular cloning. Our PCR-based method uses either bacterial artificial chromosomes or human genomic DNA as templates with {alpha}-satellite-specific primers. Here we demonstrate the production of fluorochrome-labeled DNA repeat probes specific for human chromosomes 17 and 18 in just a few days without the need for highly specialized equipment and without the limitation to only a few fluorochrome labels.

  3. The Role of Non-specific and Specific Immune Systems in Poultry against Newcastle Disease

    Directory of Open Access Journals (Sweden)

    Dyah Ayu Hewajuli

    2015-09-01

    Full Text Available Newcastle disease (ND is caused by avian paramyxovirus-1 which belong to Avulavirus genus and Paramyxoviridae family. The birds have abnormalities in humoral (bursa fabricius and cellular (thymus and spleen lymphoid organs. Lesions decrease the immune system. Immune system consists of non-specific and specific immune systems. The main components of non-specific immunity are physical and chemical barrier (feather and skin or mucosa, phagocytic cells (macrophages and natural killer, protein complement and the mediator of inflammation and cytokines. Interferons (IFNs belong to a group of cytokines that play a major role in the nonspecific or innate (natural immunity. The virulent ND virus encodes protein of V gene can be suppressed IFN type I. This leads to non-specific immune system fail to respond to the virulent strains resulting in severe pathogenicity. The defense mechanism of the host is replaced by specific immunity (adaptive immunity when natural immunity fails to overcome the infection. The specific immune system consists of humoral mediated immunity (HMI and cell-mediated immunity (CMI. The cells of immune system that react specifically with the antigen are B lymphocytes producing the antibodies, T lymphocytes that regulate the synthesis of antibodies and T cells as effector or the direct cytotoxic cells. Both non-specific and specific immunities are complementary against the invasion of ND virus in the birds. The objective of this article is to discuss the role of non specific and specific immune system in ND.

  4. Reduced goal specificity is associated with reduced memory specificity in depressed adults.

    Science.gov (United States)

    Belcher, Jessica; Kangas, Maria

    2014-01-01

    Models of autobiographical memory suggest a close association between memories, future imagination and setting specific personal goals. However this association has yet to be tested with depressed individuals. The aim of this study was to examine whether the specificity of remembering past and imagining future personal events is associated with the specificity of approach and avoidance goals in depressed individuals. Two samples comprising adults who met criteria for major depressive disorder (MDD; N=30) and adults who had no prior history or current depression (N=30) completed autobiographical memory and future event tests, and a personal goal task. In the depressed sample, the specificity with which participants remembered the past was significantly associated with the specificity with which they generated future goals. The depressed sample also elicited fewer specific approach and avoidance goals compared to the non-depressed sample. These findings suggest that an overgeneral memory deficit extends to impairments in goal specificity.

  5. [Specific features of written medical English language].

    Science.gov (United States)

    Zorica, Antić

    2010-01-01

    Medical language is a special language used by experts in mutual communication. One of the characteristics of medical discourse community is writing research papers in English. When writing a research paper, it is necessary to apply appropriate rules based on the IMRAD structure. This structure has its own clearly defined style which is clear, precise, exact and objective. This style of writing reflects the characteristics of medical language. The most frequent problems that doctors encounter in this context predominantly refer to linguistic elements of English. Namely, a very common dilemma is related to the use of English tenses in specific parts of the IMRAD structure. Another problem is the use of specific expressions for introducing a research, presenting results and methods used, developing discussion and stating conclusions. This work aims to point out the specific style and structure of medical research papers in English with an outline of the linguistic elements used in this context.

  6. Analysis on judo specific and unspecific test

    Directory of Open Access Journals (Sweden)

    Luis Santos Rodríguez

    2012-07-01

    Full Text Available In specialized literature it is not easy to find test which give us information about judokas developing their specific kinetic habits. Although judo is included in sociomotor sports (Parlebas, 2001, judokas physical fitness is normally evaluated through test which come from other type of sports, mainly from phychomotor sports. At the same time, batteries as the battery Eurofit (Conseil de L´Europe, 1989, are use to evaluate perceptive-motors aspects. In this article, we describe some of the tests found in specific western bibliography. Once finished this analysis, we mention the need of creating test which value the reality of competition confrontation more specifically, and which can be reliable, objective and reproductive (López and cols. 2004 being easy and useful to the trainer as well.

  7. Transactions in domain-specific information systems

    Science.gov (United States)

    Zacek, Jaroslav

    2017-07-01

    Substantial number of the current information system (IS) implementations is based on transaction approach. In addition, most of the implementations are domain-specific (e.g. accounting IS, resource planning IS). Therefore, we have to have a generic transaction model to build and verify domain-specific IS. The paper proposes a new transaction model for domain-specific ontologies. This model is based on value oriented business process modelling technique. The transaction model is formalized by the Petri Net theory. First part of the paper presents common business processes and analyses related to business process modeling. Second part defines the transactional model delimited by REA enterprise ontology paradigm and introduces states of the generic transaction model. The generic model proposal is defined and visualized by the Petri Net modelling tool. Third part shows application of the generic transaction model. Last part of the paper concludes results and discusses a practical usability of the generic transaction model.

  8. Testing Library Specifications by Verifying Conformance Tests

    DEFF Research Database (Denmark)

    Kiniry, Joseph Roland; Zimmerman, Daniel; Hyland, Ralph

    2012-01-01

    the Formal CTD Process (FCTD), to address this problem. Although FCTD is primarily targeted toward those who write Java libraries (or specifications for existing Java libraries) using the Java Modeling Language (JML), its techniques are broadly applicable. The key to FCTD is its novel usage of library......Formal specifications of standard libraries are necessary when statically verifying software that uses those libraries. Library specifications must be both correct, accurately reflecting library behavior, and useful, describing library behavior in sufficient detail to allow static verification...... of client programs. Specication and verification researchers regularly face the question of whether the library specications we use are correct and useful, and we have collectively provided no good answers. Over the past few years we have created and refined a software engineering process, which we call...

  9. Functional specificity of rat vibrissal primary afferents.

    Science.gov (United States)

    Lucianna, Facundo A; Farfán, Fernando D; Pizá, Gabriel A; Albarracín, Ana L; Felice, Carmelo J

    2016-06-01

    In this study, we propose to analyze the peripheral vibrissal system specificity through its neuronal responses. Receiver operating characteristics (ROC) curve analyses were used, which required the implementation of a binary classifier (artificial neural network) trained to identify the applied stimulus. The training phase consisted of the observation of a predetermined amount of vibrissal sweeps on two surfaces of different texture and similar roughness. Our results suggest that the specificity of the peripheral vibrissal system easily permits the discrimination between perceived stimuli, quantified through neuronal responses, and that it can be evaluated through an ROC curve analysis. We found that such specificity makes a linear binary classifier capable of detecting differences between stimuli with five sweeps at most.

  10. Polysaccharides in colon-specific drug delivery.

    Science.gov (United States)

    Sinha, V R; Kumria, R

    2001-08-14

    Natural polysaccharides are now extensively used for the development of solid dosage forms for delivery of drug to the colon. The rationale for the development of a polysaccharide based delivery system for colon is the presence of large amounts of polysaccharidases in the human colon as the colon is inhabited by a large number and variety of bacteria which secrete many enzymes e.g. beta-D-glucosidase, beta-D-galactosidase, amylase, pectinase, xylanase, beta-D-xylosidase, dextranase, etc. Various major approaches utilizing polysaccharides for colon-specific delivery are fermentable coating of the drug core, embedding of the drug in biodegradable matrix, formulation of drug-saccharide conjugate (prodrugs). A large number of polysaccharides have already been studied for their potential as colon-specific drug carrier systems, such as chitosan, pectin, chondroitin sulphate, cyclodextrin, dextrans, guar gum, inulin, amylose and locust bean gum. Recent efforts and approaches exploiting these polysaccharides in colon-specific drug delivery are discussed.

  11. Spent nuclear fuel project product specification

    Energy Technology Data Exchange (ETDEWEB)

    Pajunen, A.L.

    1998-01-30

    Product specifications are limits and controls established for each significant parameter that potentially affects safety and/or quality of the Spent Nuclear Fuel (SNF) packaged for transport to dry storage. The product specifications in this document cover the spent fuel packaged in MultiCanister Overpacks (MCOs) to be transported throughout the SNF Project. The SNF includes N Reactor fuel and single-pass reactor fuel. The FRS removes the SNF from the storage canisters, cleans it, and places it into baskets. The MCO loading system places the baskets into MCO/Cask assembly packages. These packages are then transferred to the Cold Vacuum Drying (CVD) Facility. After drying at the CVD Facility, the MCO cask packages are transferred to the Canister Storage Building (CSB), where the MCOs are removed from the casks, staged, inspected, sealed (by welding), and stored until a suitable permanent disposal option is implemented. The key criteria necessary to achieve these goals are documented in this specification.

  12. Computational network design from functional specifications

    KAUST Repository

    Peng, Chi Han

    2016-07-11

    Connectivity and layout of underlying networks largely determine agent behavior and usage in many environments. For example, transportation networks determine the flow of traffic in a neighborhood, whereas building floorplans determine the flow of people in a workspace. Designing such networks from scratch is challenging as even local network changes can have large global effects. We investigate how to computationally create networks starting from only high-level functional specifications. Such specifications can be in the form of network density, travel time versus network length, traffic type, destination location, etc. We propose an integer programming-based approach that guarantees that the resultant networks are valid by fulfilling all the specified hard constraints and that they score favorably in terms of the objective function. We evaluate our algorithm in two different design settings, street layout and floorplans to demonstrate that diverse networks can emerge purely from high-level functional specifications.

  13. Organelle-Specific Initiation of Autophagy.

    Science.gov (United States)

    Sica, Valentina; Galluzzi, Lorenzo; Bravo-San Pedro, José Manuel; Izzo, Valentina; Maiuri, Maria Chiara; Kroemer, Guido

    2015-08-20

    Autophagy constitutes a prominent mechanism through which eukaryotic cells preserve homeostasis in baseline conditions and in response to perturbations of the intracellular or extracellular microenvironment. Autophagic responses can be relatively non-selective or target a specific subcellular compartment. At least in part, this depends on the balance between the availability of autophagic substrates ("offer") and the cellular need of autophagic products or functions for adaptation ("demand"). Irrespective of cargo specificity, adaptive autophagy relies on a panel of sensors that detect potentially dangerous cues and convert them into signals that are ultimately relayed to the autophagic machinery. Here, we summarize the molecular systems through which specific subcellular compartments-including the nucleus, mitochondria, plasma membrane, reticular apparatus, and cytosol-convert homeostatic perturbations into an increased offer of autophagic substrates or an accrued cellular demand for autophagic products or functions. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. RESEARCH ON DIALECT-SPECIFIC LEXICON ADAPTATION

    Institute of Scientific and Technical Information of China (English)

    Liu Linquan

    2008-01-01

    With respect m dialectal-accented speech recognition, dialect-specific lexicon adaptation is an indispensable component to improve the performance. In this paper, a phone-based confusion matrix is adopted to obtain dialect-specific pronunciation variants. A weighting method, improved context-dependent weighting (ICDW) is proposed to characterize the pronunciation probability precisely by taking both the surface form left-context-dependency and the base form left-context-dependency into account. To make a much robust lexicon, a pruning criterion, syllable-dependent pruning (SDP), is also proposed which achieves the most effective result. In summary, the dialect-specific dialect adaptation reduces a 2.9% absolute and a 3.6% absolute in syllable error rate (SER) respectively on read speech and spontaneous speech from Shanghai-accented speakers.

  15. Specificity in Legume-Rhizobia Symbioses.

    Science.gov (United States)

    Andrews, Mitchell; Andrews, Morag E

    2017-03-26

    Most species in the Leguminosae (legume family) can fix atmospheric nitrogen (N₂) via symbiotic bacteria (rhizobia) in root nodules. Here, the literature on legume-rhizobia symbioses in field soils was reviewed and genotypically characterised rhizobia related to the taxonomy of the legumes from which they were isolated. The Leguminosae was divided into three sub-families, the Caesalpinioideae, Mimosoideae and Papilionoideae. Bradyrhizobium spp. were the exclusive rhizobial symbionts of species in the Caesalpinioideae, but data are limited. Generally, a range of rhizobia genera nodulated legume species across the two Mimosoideae tribes Ingeae and Mimoseae, but Mimosa spp. show specificity towards Burkholderia in central and southern Brazil, Rhizobium/Ensifer in central Mexico and Cupriavidus in southern Uruguay. These specific symbioses are likely to be at least in part related to the relative occurrence of the potential symbionts in soils of the different regions. Generally, Papilionoideae species were promiscuous in relation to rhizobial symbionts, but specificity for rhizobial genus appears to hold at the tribe level for the Fabeae (Rhizobium), the genus level for Cytisus (Bradyrhizobium), Lupinus (Bradyrhizobium) and the New Zealand native Sophora spp. (Mesorhizobium) and species level for Cicer arietinum (Mesorhizobium), Listia bainesii (Methylobacterium) and Listia angolensis (Microvirga). Specificity for rhizobial species/symbiovar appears to hold for Galega officinalis (Neorhizobium galegeae sv. officinalis), Galega orientalis (Neorhizobium galegeae sv. orientalis), Hedysarum coronarium (Rhizobium sullae), Medicago laciniata (Ensifer meliloti sv. medicaginis), Medicago rigiduloides (Ensifer meliloti sv. rigiduloides) and Trifolium ambiguum (Rhizobium leguminosarum sv. trifolii). Lateral gene transfer of specific symbiosis genes within rhizobial genera is an important mechanism allowing legumes to form symbioses with rhizobia adapted to particular soils

  16. Disease specific productivity of american cancer hospitals.

    Directory of Open Access Journals (Sweden)

    Jeffery A Goldstein

    Full Text Available Research-oriented cancer hospitals in the United States treat and study patients with a range of diseases. Measures of disease specific research productivity, and comparison to overall productivity, are currently lacking.Different institutions are specialized in research of particular diseases.To report disease specific productivity of American cancer hospitals, and propose a summary measure.We conducted a retrospective observational survey of the 50 highest ranked cancer hospitals in the 2013 US News and World Report rankings. We performed an automated search of PubMed and Clinicaltrials.gov for published reports and registrations of clinical trials (respectively addressing specific cancers between 2008 and 2013. We calculated the summed impact factor for the publications. We generated a summary measure of productivity based on the number of Phase II clinical trials registered and the impact factor of Phase II clinical trials published for each institution and disease pair. We generated rankings based on this summary measure.We identified 6076 registered trials and 6516 published trials with a combined impact factor of 44280.4, involving 32 different diseases over the 50 institutions. Using a summary measure based on registered and published clinical trails, we ranked institutions in specific diseases. As expected, different institutions were highly ranked in disease-specific productivity for different diseases. 43 institutions appeared in the top 10 ranks for at least 1 disease (vs 10 in the overall list, while 6 different institutions were ranked number 1 in at least 1 disease (vs 1 in the overall list.Research productivity varies considerably among the sample. Overall cancer productivity conceals great variation between diseases. Disease specific rankings identify sites of high academic productivity, which may be of interest to physicians, patients and researchers.

  17. Shape configuration and category-specificity

    DEFF Research Database (Denmark)

    Gerlach, Christian; Law, Ian; Paulson, Olaf B

    2006-01-01

    and fragmented drawings. We also examined whether fragmentation had different impact on the recognition of natural objects and artefacts and found that recognition of artefacts was more affected by fragmentation than recognition of natural objects. Thus, the usual finding of an advantage for artefacts...... a recent account of category-specificity and lends support to the notion that category-specific impairments can occur for both natural objects and artefacts following damage to pre-semantic stages in visual object recognition. The implications of the present findings are discussed in relation to theories...

  18. Clojure for domain-specific languages

    CERN Document Server

    Kelker, Ryan D

    2013-01-01

    An example-oriented approach to develop custom domain-specific languages.If you've already developed a few Clojure applications and wish to expand your knowledge on Clojure or domain-specific languages in general, then this book is for you. If you're an absolute Clojure beginner, then you may only find the detailed examples of the core Clojure components of value. If you've developed DSLs in other languages, this Lisp and Java-based book might surprise you with the power of Clojure.

  19. Primordial Germ Cell Specification and Migration.

    Science.gov (United States)

    Marlow, Florence

    2015-01-01

    Primordial germ cells are the progenitor cells that give rise to the gametes. In some animals, the germline is induced by zygotic transcription factors, whereas in others, primordial germ cell specification occurs via inheritance of maternally provided gene products known as germ plasm. Once specified, the primordial germ cells of some animals must acquire motility and migrate to the gonad in order to survive. In all animals examined, perinuclear structures called germ granules form within germ cells. This review focuses on some of the recent studies, conducted by several groups using diverse systems, from invertebrates to vertebrates, which have provided mechanistic insight into the molecular regulation of germ cell specification and migration.

  20. Biofilm-specific antibiotic tolerance and resistance.

    Science.gov (United States)

    Olsen, I

    2015-05-01

    Biofilms are heterogeneous structures composed of bacterial cells surrounded by a matrix and attached to solid surfaces. The bacteria here are 100 to 1,000 times more tolerant to antimicrobials than corresponding planktonic cells. Biofilms can be difficult to eradicate when they cause biofilm-related diseases, e.g., implant infections, cystic fibrosis, urinary tract infections, and periodontal diseases. A number of phenotypic features of the biofilm can be involved in biofilm-specific tolerance and resistance. Little is known about the molecular mechanisms involved. The current review deals with both phenotypic and molecular mechanisms of biofilm-specific antibiotic tolerance and resistance.

  1. Domain-Specific Languages for Enterprise Systems

    DEFF Research Database (Denmark)

    Andersen, Jesper; Bahr, Patrick; Henglein, Fritz;

    2014-01-01

    The process-oriented event-driven transaction systems (POETS) architecture introduced by Henglein et al. is a novel software architecture for enterprise resource planning (ERP) systems. POETS employs a pragmatic separation between (i) transactional data, that is, what has happened; (ii) reports......, that is, what can be derived from the transactional data; and (iii) contracts, that is, which transactions are expected in the future. Moreover, POETS applies domain-specific languages (DSLs) for specifying reports and contracts, in order to enable succinct declarative specifications as well as rapid...

  2. From UML Specification into FPGA Implementation

    Directory of Open Access Journals (Sweden)

    Grzegorz Bazydlo

    2014-01-01

    Full Text Available In the paper a method of using the Unified Modeling Language for specification of digital systems, especially logic controllers, is presented. The proposed method is based mainly on the UML state machine diagrams and uses Hierarchical Concurrent Finite State Machines (HCFSMs as a temporary model. The paper shows a way to transform the UML diagrams, expressed in XML language, to the form that is acceptable by reconfigurable FPGAs (Field Programmable Gate Arrays. The UML specification is used to generate an effective program in Hardware Description Languages (HDLs, especially Verilog.

  3. [Development of domain specific search engines].

    Science.gov (United States)

    Takai, T; Tokunaga, M; Maeda, K; Kaminuma, T

    2000-01-01

    As cyber space exploding in a pace that nobody has ever imagined, it becomes very important to search cyber space efficiently and effectively. One solution to this problem is search engines. Already a lot of commercial search engines have been put on the market. However these search engines respond with such cumbersome results that domain specific experts can not tolerate. Using a dedicate hardware and a commercial software called OpenText, we have tried to develop several domain specific search engines. These engines are for our institute's Web contents, drugs, chemical safety, endocrine disruptors, and emergent response for chemical hazard. These engines have been on our Web site for testing.

  4. Automated effect-specific mammographic pattern measures

    DEFF Research Database (Denmark)

    Raundahl, Jakob; Loog, Marco; Pettersen, Paola;

    2008-01-01

    We investigate the possibility to develop methodologies for assessing effect specific structural changes of the breast tissue using a general statistical machine learning framework. We present an approach of obtaining objective mammographic pattern measures quantifying a specific biological effect......, such as hormone replacement therapy (HRT). We compare results using this approach to using standard density measures. We show that the proposed method can quantify both age related effects and effects caused by HRT. Age effects are significantly detected by our method where standard methodologies fail...

  5. Spent Nuclear Fuel (SNF) Project Product Specification

    Energy Technology Data Exchange (ETDEWEB)

    PAJUNEN, A.L.

    2000-12-07

    The process for removal of Spent Nuclear Fuel (SNF) from the K Basins has been divided into major sub-systems. The Fuel Retrieval System (FRS) removes fuel from the existing storage canisters, cleans it, and places it into baskets. The multi-canister overpack (MCO) loading system places the baskets into an MCO that has been pre-loaded in a cask. The cask, containing a loaded MCO, is then transferred to the Cold Vacuum Drying (CVD) Facility. After drying at the CVD Facility, the cask, and MCO, are transferred to the Canister Storage Building (CSB), where the MCO is removed from the cask, staged, inspected, sealed (by welding), and stored until a suitable permanent disposal option is implemented. The purpose of this document is to specify the process related characteristics of an MCO at the interface between major process systems. The characteristics are derived from the primary technical documents that form the basis for safety analysis and design calculations. This document translates the calculation assumptions into implementation requirements and describes the method of verifying that the requirement is achieved. These requirements are used to define validation test requirements and describe requirements that influence multiple sub-project safety analysis reports. This product specification establishes limits and controls for each significant process parameter at interfaces between major sub-systems that potentially affect the overall safety and/or quality of the SNF packaged for processing, transport, and interim dry storage. The product specifications in this document cover the SNF packaged in MCOs to be transported throughout the SNF Project. The description of the product specifications are organized in the document as follows: Section 2.0--Summary listing of product specifications at each major sub-system interface. Section 3.0--Summary description providing guidance as to how specifications are complied with by equipment design or processing within a major

  6. Automated effect-specific mammographic pattern measures.

    Science.gov (United States)

    Raundahl, Jakob; Loog, Marco; Pettersen, Paola; Tanko, Laszlo B; Nielsen, Mads

    2008-08-01

    We investigate the possibility to develop methodologies for assessing effect specific structural changes of the breast tissue using a general statistical machine learning framework. We present an approach of obtaining objective mammographic pattern measures quantifying a specific biological effect, such as hormone replacement therapy (HRT). We compare results using this approach to using standard density measures. We show that the proposed method can quantify both age related effects and effects caused by HRT. Age effects are significantly detected by our method where standard methodologies fail. The separation of HRT subpopulations using our approach is comparable to the best methodology, which is interactive.

  7. Specific learning processes and indigenous teacher training

    Directory of Open Access Journals (Sweden)

    Adir Casaro Nascimento

    2012-11-01

    Full Text Available Indigenous teacher formation and the issue of specific learning processes, as a right of the indigenous peoples derived from the 1988 Constitution, aim at the re-signification of pedagogical practices in specific socio-cultural contexts and at the visibility of indigenous education. Taking indigenous children as a reference, or rather, the agents that produce knowledge within the context of their particularities and territorialities, the essay points to the necessity of constructing new theoretical bases and a pedagogy that gives visibility to other local epistemic logics produced by “power coloniality”. They are different from the dominant Western logic in the process of training indigenous educators.

  8. Non-specific ileojejunitis in Arequipa, Peru.

    Science.gov (United States)

    Perea, V D; Fernan-Zegarra, L; Cruz, V M; Ballon, R; Picoaga, J L

    1978-09-09

    Non-specific ileojejunitis, characterised by mild to moderate structural changes in the intestinal mucosa and often associated with asymptomatic malabsorption, is known to occur in the tropics and in countries with hot climates. This acquired condition is probably related to environmental factors, and some consider it to be a subclinical form of tropical sprue. Changes in the intestinal mucosa typical of non-specific ileojejunitis were found in 10 indigenous Indians as well as 5 people of Latin stock living in the Southern Peruvian sierra, where tropical sprue has not as yet been demonstrated and which has a temperate climate.

  9. Specifications from a biotechnology industry perspective.

    Science.gov (United States)

    Garnick, R L

    1997-01-01

    The emergence of new analytical technology and the production of pharmaceuticals for a global market in a cost-effective manner necessitate the establishment of worldwide specifications that are appropriate for the product and the manufacturing process. This requires a thorough knowledge of the protein and control of the systems that produce it as well as an understanding of the accuracy and precision of the assays used for testing. Harmonization of specifications among the worldwide regulatory authorities is critical for the future development of new pharmaceuticals. A continuing dialogue between industry and regulators to achieve this goal needs to be encouraged and supported.

  10. Shape configuration and category-specificity

    DEFF Research Database (Denmark)

    Gerlach, Christian; Law, I; Paulson, Olaf B.

    2006-01-01

    in difficult object decision tasks, which is also found in the present experiments with outlines, is reversed when the stimuli are fragmented. This interaction between category (natural versus artefacts) and stimulus type (outlines versus fragmented forms) is in accordance with predictions derived from...... a recent account of category-specificity and lends support to the notion that category-specific impairments can occur for both natural objects and artefacts following damage to pre-semantic stages in visual object recognition. The implications of the present findings are discussed in relation to theories...

  11. A Class-Specific Optimizing Compiler

    Directory of Open Access Journals (Sweden)

    Michael D. Sharp

    1993-01-01

    Full Text Available Class-specific optimizations are compiler optimizations specified by the class implementor to the compiler. They allow the compiler to take advantage of the semantics of the particular class so as to produce better code. Optimizations of interest include the strength reduction of class:: array address calculations, elimination of large temporaries, and the placement of asynchronous send/recv calls so as to achieve computation/communication overlap. We will outline our progress towards the implementation of a C++ compiler capable of incorporating class-specific optimizations.

  12. Engineered waste-package-system design specification

    Energy Technology Data Exchange (ETDEWEB)

    1983-05-01

    This report documents the waste package performance requirements and geologic and waste form data bases used in developing the conceptual designs for waste packages for salt, tuff, and basalt geologies. The data base reflects the latest geotechnical information on the geologic media of interest. The parameters or characteristics specified primarily cover spent fuel, defense high-level waste, and commercial high-level waste forms. The specification documents the direction taken during the conceptual design activity. A separate design specification will be developed prior to the start of the preliminary design activity.

  13. Domain Specific Language Support for Exascale

    Energy Technology Data Exchange (ETDEWEB)

    Sadayappan, Ponnuswamy [The Ohio State Univ., Columbus, OH (United States)

    2017-02-24

    Domain-Specific Languages (DSLs) offer an attractive path to Exascale software since they provide expressive power through appropriate abstractions and enable domain-specific optimizations. But the advantages of a DSL compete with the difficulties of implementing a DSL, even for a narrowly defined domain. The DTEC project addresses how a variety of DSLs can be easily implemented to leverage existing compiler analysis and transformation capabilities within the ROSE open source compiler as part of a research program focusing on Exascale challenges. The OSU contributions to the DTEC project are in the area of code generation from high-level DSL descriptions, as well as verification of the automatically-generated code.

  14. NEEDS ANALYSIS IN ENGLISH FOR SPECIFIC PURPOSES

    Directory of Open Access Journals (Sweden)

    SIMION MINODORA OTILIA

    2015-03-01

    Full Text Available Needs analysis is considered to be the cornerstone of English for Specific Purposes(ESP. The concept of Needs analysis has been different along the decades. At the initial stages of ESP( the 1960s and early 1970s, needs analysis consisted in assessing the communicative needs of the learners and the techniques of achieving specific teaching objectives. Nowadays, the tasks of needs analysis is much more complex: it aims at collecting information about the learners and at defining the target situation and environment of studying ESP.

  15. CASL- The Common Algebraic Specification Language- Summary

    DEFF Research Database (Denmark)

    Haxthausen, Anne

    1997-01-01

    This Summary is the basis for the Design Proposal [LD97b] for CASL, the Common Algebraic Specification Language, prepared by the Language Design Task Group of CoFI, the Common Framework Initiative. It gives the abstract syntax, and informally describes its intended semantics. It is accompanied...... for approval to the sponsoring IFIP Working Group on Foundations of System Specification, WG 1.3. It received tentative approval, together with a referees' report recommending the reconsideration of some elements of the design [IFI97]; a response has already been made [LD97a]. The present version...

  16. Effect of MSTN Propeptide and shRNA Co-expression Vector on Proliferation of Skeletal Muscle Satellite Cells

    Institute of Scientific and Technical Information of China (English)

    Feng Lin-he; Wang Xin; Lu Ming; Tong Hui-li; Li Shu-feng; Yan Yun-qin

    2014-01-01

    Myostatin (MSTN) is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro-MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly (P<0.05). After knocked down MSTN gene, P21 expression decreased (P<0.05), but CDK2 gene expression increased (P<0.05). These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.

  17. Silencing of SOX12 by shRNA suppresses migration, invasion and proliferation of breast cancer cells

    Science.gov (United States)

    Ding, Hanzhi; Quan, Hong; Yan, Weiguo; Han, Jing

    2016-01-01

    Sex determining region Y-box protein 12 (SOX12) is essential for embryonic development and cell-fate determination. The role of SOX12 in tumorigenesis of breast cancer is not well-understood. Here, we found that SOX12 mRNA expression was up-regulated in human breast cancer tissues. To clarify the roles of SOX12 in breast cancer, we used lentiviral shRNAs to suppress its expression in two breast cancer cells with relatively higher expression of SOX12 (BT474 and MCF-7). Our findings strongly suggested that SOX12 was critical for cell migration and invasion of breast cancer cells. We found that silencing of SOX12 significantly decreased the mRNA and protein levels of MMP9 and Twist, while notably increased E-cadherin. Moreover, SOX12 knockdown significantly inhibited the proliferation of breast cancer cells in vitro and the growth of xenograft tumours in vivo. Flow cytometry analysis revealed that breast cancer cells with SOX12 knockdown showed cell cycle arrest and decreased mRNA and protein levels of proliferating cell nuclear antigen (PCNA), CDK2 and Cyclin D1. Taken together, SOX12 plays an important role in growth inhibition through cell-cycle arrest, as well as migration and invasion of breast cancer cells. PMID:27582508

  18. Cystatin M/E knockdown by lentiviral delivery of shRNA impairs epidermal morphogenesis of human skin equivalents

    NARCIS (Netherlands)

    Jansen, P.A.M.; Bogaard, E.H. van den; Kersten, F.F.J.; Oostendorp, C.; Vlijmen-Willems, I.M. van; Oji, V.; Traupe, H.; Hennies, H.C.; Schalkwijk, J.; Zeeuwen, P.L.J.M.

    2012-01-01

    The protease inhibitor cystatin M/E (CST6) regulates a biochemical pathway involved in stratum corneum homeostasis, and its deficiency in mice causes ichthyosis and neonatal lethality. Cystatin M/E deficiency has not been described in humans so far, and we did not detect disease-causing mutations in

  19. SHRNA-MEDIATED SILENCING OF AS3MT EXPRESSION MODULATES THE CAPACITY OF HEPG2 CELLS TO METHYLATE INORGANIC ARSENIC

    Science.gov (United States)

    Several methyltransferases have been linked to the oxidative methylation of inorganic arsenic (iAs) in mammalian cells. However, the relative contributions of these enzymes to the overall capacity of cells to methylate iAs have not been characterized. Arsenic (+3 oxidation state)...

  20. Measuring Language-Specific Phonetic Settings

    Science.gov (United States)

    Mennen, Ineke; Scobbie, James M.; de Leeuw, Esther; Schaeffler, Sonja; Schaeffler, Felix

    2010-01-01

    While it is well known that languages have different phonemes and phonologies, there is growing interest in the idea that languages may also differ in their "phonetic setting". The term "phonetic setting" refers to a tendency to make the vocal apparatus employ a language-specific habitual configuration. For example, languages may differ in their…

  1. Age-Specific Correlates of Child Growth

    NARCIS (Netherlands)

    M. Rieger (Matthias); S.K. Trommlerová (Sofia Karina)

    2016-01-01

    textabstractGrowth faltering describes a widespread phenomenon that height- and weight-for-age of children in developing countries collapse rapidly in the first two years of life. We study age-specific correlates of child nutrition using Demographic and Health Surveys from 56 developing countries to

  2. Specification of the somatic musculature in Drosophila.

    Science.gov (United States)

    Dobi, Krista C; Schulman, Victoria K; Baylies, Mary K

    2015-01-01

    The somatic muscle system formed during Drosophila embryogenesis is required for larvae to hatch, feed, and crawl. This system is replaced in the pupa by a new adult muscle set, responsible for activities such as feeding, walking, and flight. Both the larval and adult muscle systems are comprised of distinct muscle fibers to serve these specific motor functions. In this way, the Drosophila musculature is a valuable model for patterning within a single tissue: while all muscle cells share properties such as the contractile apparatus, properties such as size, position, and number of nuclei are unique for a particular muscle. In the embryo, diversification of muscle fibers relies first on signaling cascades that pattern the mesoderm. Subsequently, the combinatorial expression of specific transcription factors leads muscle fibers to adopt particular sizes, shapes, and orientations. Adult muscle precursors (AMPs), set aside during embryonic development, proliferate during the larval phases and seed the formation of the abdominal, leg, and flight muscles in the adult fly. Adult muscle fibers may either be formed de novo from the fusion of the AMPs, or are created by the binding of AMPs to an existing larval muscle. While less is known about adult muscle specification compared to the larva, expression of specific transcription factors is also important for its diversification. Increasingly, the mechanisms required for the diversification of fly muscle have found parallels in vertebrate systems and mark Drosophila as a robust model system to examine questions about how diverse cell types are generated within an organism.

  3. Designing specificity of protein-substrate interactions

    NARCIS (Netherlands)

    Coluzza, I.; Frenkel, D.

    2004-01-01

    One of the key properties of biological molecules is that they can bind strongly to certain substrates yet interact only weakly with the very large number of other molecules that they encounter. Using a simple lattice model, we test several methods to design molecule-substrate binding specificity. W

  4. 16 CFR 1209.35 - Product specification.

    Science.gov (United States)

    2010-01-01

    ... INTERIM SAFETY STANDARD FOR CELLULOSE INSULATION Certification § 1209.35 Product specification. (a..., the cellulose insulation may be manufactured and distributed in commerce, subject to the provisions of... factor which is likely to affect the ability of the cellulose insulation to meet the standard, that...

  5. Specification and Configuration of Customized Complex Products

    NARCIS (Netherlands)

    Afsarmanesh, H.; Shafahi, M.

    2013-01-01

    This paper addresses the design of an information system for specification of complex configured products, such as the solar power plants or large intelligent buildings, which by nature are designed, constructed, installed, operated, and maintained through virtual consortium of enterprises. In other

  6. Digital PET compliance to EARL accreditation specifications

    NARCIS (Netherlands)

    Koopman, Daniëlle; Groot Koerkamp, Maureen; Jager, Pieter L.; Arkies, Hester; Knollema, Siert; Slump, Cornelis H.; Sanches, Pedro G.; van Dalen, Jorn A.

    2017-01-01

    Background: Our aim was to evaluate if a recently introduced TOF PET system with digital photon counting technology (Philips Healthcare), potentially providing an improved image quality over analogue systems, can fulfil EANM research Ltd (EARL) accreditation specifications for tumour imaging with

  7. Specific Ion Effects in Cholesterol Monolayers

    Directory of Open Access Journals (Sweden)

    Teresa Del Castillo-Santaella

    2016-05-01

    Full Text Available The interaction of ions with interfaces and, in particular, the high specificity of these interactions to the particular ions considered, are central questions in the field of surface forces. Here we study the effect of different salts (NaI, NaCl, CaCl2 and MgCl2 on monolayers made of cholesterol molecules, both experimentally (surface area vs. lateral pressure isotherms measured by a Langmuir Film Balance and theoretically (molecular dynamics (MD all-atomic simulations. We found that surface isotherms depend, both quantitatively and qualitatively, on the nature of the ions by altering the shape and features of the isotherm. In line with the experiments, MD simulations show clear evidences of specific ionic effects and also provide molecular level details on ion specific interactions with cholesterol. More importantly, MD simulations show that the interaction of a particular ion with the surface depends strongly on its counterion, a feature ignored so far in most theories of specific ionic effects in surface forces.

  8. Chromosome-specific families in Vibrio genomes

    Directory of Open Access Journals (Sweden)

    Oksana eLukjancenko

    2014-03-01

    Full Text Available We have compared chromosome-specific genes in a set of 18 finished Vibrio genomes, and, in addition, also calculated the pan- and core-genomes from a data set of more than 250 draft Vibrio genome sequences. These genomes come from 9 known species and 2 unknown species. Within the finished chromosomes, we find a core set of 1269 encoded protein families for chromosome 1, and a core of 252 encoded protein families for chromosome 2. Many of these core proteins are also found in the draft genomes (although which chromosome they are located on is unknown. Of the chromosome specific core protein families, 1169 and 153 are uniquely found in chromosomes 1 and 2, respectively. Gene ontology (GO terms for each of the protein families were determined, and the different sets for each chromosome were compared. A total of 363 different `Molecular Function` GO categories were found for chromosome 1 specific protein families, and these include several broad activities: pyridoxine 5' phosphate synthetase, glucosylceramidase, heme transport, DNA ligase, amino acid binding, and ribosomal components; in contrast, chromosome 2 specific protein families have only 66 Molecular Function GO terms and include many membrane-associated activities, such as ion channels, transmembrane transporters, and electron transport chain proteins. Thus, it appears that whilst there are many 'housekeeping systems' encoded in chromosome 1, there are far fewer core functions found in chromosome 2. However, the presence of many membrane-associated encoded proteins in chromosome 2 is surprising.

  9. Development of the Specific Adaptation Mobility Cane.

    Science.gov (United States)

    Arrington, S.

    1995-01-01

    A travel cane was adapted for use by a 10-year-old boy with cortical blindness, severe mental retardation and cerebral palsy affecting his left arm and leg. The Specific Adaptation Mobility Cane utilizes the affected arm to hold the cane while leaving the other hand free for trailing walls, opening doors, carrying objects, and holding handrails.…

  10. Development of industrial variant specification systems

    DEFF Research Database (Denmark)

    Hansen, Benjamin Loer

    With globalisation and increased competition industrial companies must be prepared to satisfy individual customer needs and still stay competitive with regards to lead times, quality, and prices. These factors require companies to be better prepared to handle specification activities during order...

  11. Statistical analysis of protein kinase specificity determinants

    DEFF Research Database (Denmark)

    Kreegipuu, Andres; Blom, Nikolaj; Brunak, Søren;

    1998-01-01

    The site and sequence specificity of protein kinase, as well as the role of the secondary structure and surface accessibility of the phosphorylation sites on substrate proteins, was statistically analyzed. The experimental data were collected from the literature and are available on the World Wide...

  12. Measuring Language-Specific Phonetic Settings

    Science.gov (United States)

    Mennen, Ineke; Scobbie, James M.; de Leeuw, Esther; Schaeffler, Sonja; Schaeffler, Felix

    2010-01-01

    While it is well known that languages have different phonemes and phonologies, there is growing interest in the idea that languages may also differ in their "phonetic setting". The term "phonetic setting" refers to a tendency to make the vocal apparatus employ a language-specific habitual configuration. For example, languages may differ in their…

  13. Specific Electrostatic Molecular Recognition in Water

    DEFF Research Database (Denmark)

    Li, Ming; Hoeck, Casper; Schoffelen, Sanne;

    2016-01-01

    -bead binding assay and by 2D NMR spectroscopy. Molecular dynamics (MD) studies revealed a putative mode of interaction for this unusual electrostatic binding event. High binding specificity occurred through a combination of topological matching and electrostatic and hydrogen-bond complementarities. From MD...

  14. Specific Impulses Losses in Solid Propellant Rockets

    Science.gov (United States)

    1974-12-17

    to use the collision function form proposed by Golovin to simplify this production term: 4C><=) <P- .: Accordingly: m hence, by integration: Now, we...November 21, 1940 in Paris, Seine. VFirst Thesis. "Contribution to the Study of Specific i Impulse Loss in Solid Propellant Rockets." Second Thesis

  15. On Quality Assessment of Learning Technology Specifications

    NARCIS (Netherlands)

    Janssen, José; Berlanga, Adriana; Koper, Rob

    2012-01-01

    Janssen, J., Berlanga, A. J., & Koper, R. (2012). On Quality Assessment of Learning Technology Specifications. In J. C. Ramalho, A. Simões, & R. Queirós (Eds.). Innovations in XML Applications and Metadata Management: Advancing Technologies (pp. 78-95). Hershey, PA, USA: IGI Global. Below file

  16. Task-specificity of balance training.

    Science.gov (United States)

    Giboin, Louis-Solal; Gruber, Markus; Kramer, Andreas

    2015-12-01

    Despite much research on balance training, it is still unclear whether balance training leads to highly task-specific adaptations or rather non-specific adaptations. Hence, in this study we examined whether balance training increased performance only in the balance task that was trained or also in non-trained tasks. Forty healthy participants (28 m 12 f, 25 ± 4 years, 177 ± 10 cm, 73 ± 14 kg) were assigned to one of two training groups (TGs) or a control group. Both TGs completed six sessions over 2 weeks, only the training device differed. Before and after the training, performance in the trained task as well as in additional untrained tasks was recorded. ANOVAs showed that each TG outperformed the other groups only in the task they had trained (e.g., task trained by TG1: +225% in TG1, only +41% and +30% in TG2 and control, group*time interaction, pbalance training resulted in highly task-specific effects, no transfer even to very similar tasks was observed. Therefore, we recommend identifying and training exactly those tasks that need improvement, and test the efficacy of training programs using specific tests instead of general tests with limited functional relevance.

  17. Prostate-specific antigen (PSA) blood test

    Science.gov (United States)

    Prostate-specific antigen; Prostate cancer screening test; PSA ... special steps are needed to prepare for this test. ... Reasons for a PSA test: This test may be done to screen for prostate cancer. It is also used to follow people after prostate cancer ...

  18. The Formal Specifications for Protocols of Decoders

    Institute of Scientific and Technical Information of China (English)

    YUAN Meng-ting; WU Guo-qing; SHU Feng-di

    2004-01-01

    This paper presents a formal approach, FSPD (Formal Specifications for Protocols of Decoders), to specify decoder communication protocols. Based on axiomatic, FSPD is a precise language with which programmers could use only one suitable driver to handle various types of decoders. FSPD is helpful for programmers to get high adaptability and reusability of decoder-driver software.

  19. Diamond Drilling Specification Manual and Course Outline.

    Science.gov (United States)

    British Columbia Dept. of Education, Victoria.

    This publication presents the standards required of a person practicing diamond drilling in western Canada and provides an outline for teaching the skills and knowledge. It is divided into two parts. The Diamond Drilling Specification Manual establishes the levels of skill and knowledge required in the four certified levels of diamond drilling.…

  20. Autobiographical Memory Specificity in Dissociative Identity Disorder

    NARCIS (Netherlands)

    Huntjens, R.J.C.; Wessel, I.; Hermans, D.; Minnen, Agnes van

    2014-01-01

    A lack of adequate access to autobiographical knowledge has been related to psychopathology. More specifically, patients suffering from depression or a history of trauma have been found to be characterized by overgeneral memory, in other words, they show a relative difficulty in retrieving a specifi

  1. Isotope specific arbitrary material flow meter

    Energy Technology Data Exchange (ETDEWEB)

    Barty, Christopher P. J.; Post, John C.; Jones, Edwin

    2016-10-25

    A laser-based mono-energetic gamma-ray source is used to provide non-destructive and non-intrusive, quantitative determination of the absolute amount of a specific isotope contained within pipe as part of a moving fluid or quasi-fluid material stream.

  2. Primary Auditory Cortex Regulates Threat Memory Specificity

    Science.gov (United States)

    Wigestrand, Mattis B.; Schiff, Hillary C.; Fyhn, Marianne; LeDoux, Joseph E.; Sears, Robert M.

    2017-01-01

    Distinguishing threatening from nonthreatening stimuli is essential for survival and stimulus generalization is a hallmark of anxiety disorders. While auditory threat learning produces long-lasting plasticity in primary auditory cortex (Au1), it is not clear whether such Au1 plasticity regulates memory specificity or generalization. We used…

  3. 12 CFR 347.119 - Specific consent.

    Science.gov (United States)

    2010-01-01

    ... processing procedure. (d) Specific consent. Direct or indirect investments in or activities of foreign... control such organization as a result of a foreign investment; or (ii) A bank would be establishing a... foreign country. (1) Applicable law or practice in the foreign country where the foreign organization...

  4. Methodes for identification of specific language impairment

    Directory of Open Access Journals (Sweden)

    Toktam Maleki Shahmahmood

    2014-06-01

    Full Text Available Background and Aim: Specific language impiarment (SLI is one of the most prevalent developmental language disorders its diagnosis is a problematic issue among researchers and clinicians because of the heterogeneity of language profiles in the affected population and overlapping with other developmental language disorders. The aim of this study was to review the suggested diagnostic criteria for this disorder, controversies about these criteria and identify the most accurate diagnostic methods.Methods: Published article from 1980 to 2012 in bibliographic and publisher databases including Pubmed, Google scholar, Cochran library, Web of Science, ProQuest, Springer, Oxford, Science direct, Ovid, Iran Medex and Magiran about the diagnostic methods for discriminating preschoool children with specific language impiarment from normal developing children were reviewd in this article. These keywords were used for research: “specific language impairment”, “SLI”, “diagnosis or identification”, “standardized tests”, and “tests for language development”.Conclusion: The results of this study show inspite of agreement of researchers and clinicians about exclusionary criteria as one basic part of the diagnosis of specific language impiarment , there is no consensus about the other part, inclusionary criteria. Different studies used different inclusionary criteria which can be divided to categories of clincal judgment, discrepancy-based criteria, standardized testing, clinical markers and markers from spontaneous speech samples. Advantages, disadvantages, and clinical applicability of each diagnostic method are discussed in this article.

  5. UROKINASE MUTANT WITH BETTER FIBRIN SPECIFICITY

    Institute of Scientific and Technical Information of China (English)

    MaZhong; YuRuirong; HuaZichun; ZhuDexu

    1997-01-01

    Urokinsae (UK),one of the principal activators of plasminogen, has been widely used in the clinical therapy of thrombosis. However ,it lacks fibrin specificity ,and can lead to extensive systemic activation of plasminogen when injected into veins in large doses. On the other hand,

  6. Preliminary Anthropometric Specification for Land Vehicles

    Science.gov (United States)

    2012-05-01

    worked for QinetiQ and BAE Systems in the UK. He has provided Human Factors expertise to numerous procurement programs and currently leads the...specification be updated when sufficient data becomes available for the female Army population. 4. It is recommended that the intergenerational change in

  7. Designing specificity of protein-substrate interactions

    NARCIS (Netherlands)

    Coluzza, I.; Frenkel, D.

    2004-01-01

    One of the key properties of biological molecules is that they can bind strongly to certain substrates yet interact only weakly with the very large number of other molecules that they encounter. Using a simple lattice model, we test several methods to design molecule-substrate binding specificity.

  8. 30 CFR 22.7 - Specific requirements.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Specific requirements. 22.7 Section 22.7 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR TESTING, EVALUATION, AND... on a wood floor from a height of 3 feet. Parts that are strapped to the user may be subjected to...

  9. Teaching Case: Enterprise Architecture Specification Case Study

    Science.gov (United States)

    Steenkamp, Annette Lerine; Alawdah, Amal; Almasri, Osama; Gai, Keke; Khattab, Nidal; Swaby, Carval; Abaas, Ramy

    2013-01-01

    A graduate course in enterprise architecture had a team project component in which a real-world business case, provided by an industry sponsor, formed the basis of the project charter and the architecture statement of work. The paper aims to share the team project experience on developing the architecture specifications based on the business case…

  10. Spent Nuclear Fuel (SNF) Project Product Specification

    Energy Technology Data Exchange (ETDEWEB)

    PAJUNEN, A.L.

    2000-01-20

    This document establishes the limits and controls for the significant parameters that could potentially affect the safety and/or quality of the Spent Nuclear Fuel (SNF) packaged for processing, transport, and storage. The product specifications in this document cover the SNF packaged in Multi-Canister Overpacks to be transported throughout the SNF Project.

  11. Autobiographical memory specificity in dissociative identity disorder

    NARCIS (Netherlands)

    Huntjens, Rafaële J. C.; Wessel, Ineke; Hermans, Dirk; van Minnen, Agnes

    A lack of adequate access to autobiographical knowledge has been related to psychopathology. More specifically, patients suffering from depression or a history of trauma have been found to be characterized by overgeneral memory, in other words, they show a relative difficulty in retrieving a

  12. Autobiographical Memory Specificity in Dissociative Identity Disorder

    NARCIS (Netherlands)

    Huntjens, R.J.C.; Wessel, I.; Hermans, D.; Minnen, Agnes van

    2014-01-01

    A lack of adequate access to autobiographical knowledge has been related to psychopathology. More specifically, patients suffering from depression or a history of trauma have been found to be characterized by overgeneral memory, in other words, they show a relative difficulty in retrieving a

  13. Canine specific ELISA for coagulation factor VII

    DEFF Research Database (Denmark)

    Knudsen, Tom; Kjelgaard-Hansen, Mads; Tranholm, Mikael;

    2011-01-01

    available to date. In this study, a canine specific ELISA for measurement of FVII:Ag in plasma was developed and validated. The FVII:Ag ELISA correctly diagnosed homozygous and heterozygous hereditary FVII deficiency. Together with activity based assays, such as FVII:C, the FVII:Ag ELISA should be valuable...

  14. An Introduction to Product Specifications in China

    Institute of Scientific and Technical Information of China (English)

    Yin Li; Wang Zheng

    2010-01-01

    @@ Preface The government and enterprises in China started to pay attention to product specifications beginning with the launch of reform and, especially after the state promulgated the following two laws in 1993:the Product Quality Law (Article 15) and the Law on Protection of Consumer Rights and Interests (Articles 8, 13, 18, 22, 23, etc.).

  15. 48 CFR 836.202 - Specifications.

    Science.gov (United States)

    2010-10-01

    ... specifications. (b) During the design stage, contract architect-engineers must not use “brand name or equal” or... OF CONTRACTING CONSTRUCTION AND ARCHITECT-ENGINEER CONTRACTS Special Aspects of Contracting for... contracting officer must inform prospective architect-engineers of this requirement during the...

  16. Analyzing Languages for Specific Purposes Discourse

    Science.gov (United States)

    Bowles, Hugo

    2012-01-01

    In the last 20 years, technological advancement and increased multidisciplinarity has expanded the range of data regarded as within the scope of languages for specific purposes (LSP) research and the means by which they can be analyzed. As a result, the analytical work of LSP researchers has developed from a narrow focus on specialist terminology…

  17. 50 CFR 648.200 - Specifications.

    Science.gov (United States)

    2010-10-01

    ..., DEPARTMENT OF COMMERCE FISHERIES OF THE NORTHEASTERN UNITED STATES Management Measures for the Atlantic... (PRT) to develop and recommend the following specifications for a period of 3 years for consideration... herring fishery; (v) Total world export potential by herring producer countries; (vi) Total world...

  18. Prospects for site specific weed management

    DEFF Research Database (Denmark)

    Christensen, Svend; Rasmussen, Jesper; Pedersen, Søren Marcus;

    2014-01-01

    Research on Site Specific Weed Management (SSWM) started in the late 80's. Since that moment, considerable research has been conducted on different aspects of SSWM, from fundamental studies on the spatial ecology of weeds to the applied development and testing of new technologies for weed detection...

  19. Volatiles in inter-specific bacterial interactions

    NARCIS (Netherlands)

    Tyc, O.; Zweers, H.; De Boer, W.; Garbeva, P.V.

    2015-01-01

    The importance of volatile organic compounds for functioning of microbes is receiving increased research attention. However, to date very little is known on how inter-specific bacterial interactions effect volatiles production as most studies have been focused on volatiles produced by monocultures o

  20. Designing Nonwovens to Meet Pore Size Specifications

    Directory of Open Access Journals (Sweden)

    Glen E. Simmonds

    2007-04-01

    Full Text Available New nonwovens applications in areas such as filtration require a media designed to particular pore size specifications in the 3 to 20 micron range. The purpose of this work was to develop a basis by which to design and construct a fabric with given pore size specifications. While doing so we have provided a validation for two different mathematical models. We have also found that bicomponent spunbonded islands-in-the-sea nonwoven fabrics can be designed very precisely to achieve target pore diameters and porosity. Mathematical models can be used to develop fabric specifications in the standard manufacturing terms of basis weight and fiber diameter. Measured mean flow pore diameters for the test fabrics showed excellent correlation to targeted mean flow pore diameters for both models. The experimental fit to the Bryner model is the better of the two, but requires specification of fabric thickness in addition to basis weight and fiber diameter to achieve actual mean pore diameters that closely match target values. Experimental validation of the influence of fabric thickness on the mean flow pore diameter at constant basis weight and fiber diameter remains open for further investigation. In addition, achieving complete separation of the island and sea polymers along with unbundling of the island fibers remain areas for improvement.

  1. Ontology Assisted Formal Specification Extraction from Text

    Directory of Open Access Journals (Sweden)

    Andreea Mihis

    2010-12-01

    Full Text Available In the field of knowledge processing, the ontologies are the most important mean. They make possible for the computer to understand better the natural language and to make judgments. In this paper, a method which use ontologies in the semi-automatic extraction of formal specifications from a natural language text is proposed.

  2. Primary Auditory Cortex Regulates Threat Memory Specificity

    Science.gov (United States)

    Wigestrand, Mattis B.; Schiff, Hillary C.; Fyhn, Marianne; LeDoux, Joseph E.; Sears, Robert M.

    2017-01-01

    Distinguishing threatening from nonthreatening stimuli is essential for survival and stimulus generalization is a hallmark of anxiety disorders. While auditory threat learning produces long-lasting plasticity in primary auditory cortex (Au1), it is not clear whether such Au1 plasticity regulates memory specificity or generalization. We used…

  3. 14 CFR 91.1015 - Management specifications.

    Science.gov (United States)

    2010-01-01

    ... operations and, if different, the address that will serve as the primary point of contact for correspondence... Operations Program Management § 91.1015 Management specifications. (a) Each person conducting operations... authorizations, limitations, and certain procedures under which these operations are to be conducted, (3)...

  4. Public Address Systems. Specifications - Installation - Operation.

    Science.gov (United States)

    Palmer, Fred M.

    Provisions for public address in new construction of campus buildings (specifications, installations, and operation of public address systems), are discussed in non-technical terms. Consideration is given to microphones, amplifiers, loudspeakers and the placement and operation of various different combinations. (FS)

  5. Spent nuclear fuel project product specification

    Energy Technology Data Exchange (ETDEWEB)

    PAJUNEN, A.L.

    1999-02-25

    This document establishes the limits and controls for the significant parameters that could potentially affect the safety and/or quality of the Spent Nuclear Fuel (SNF) packaged for processing, transport, and storage. The product specifications in this document cover the SNF packaged in Multi-Canister Overpacks to be transported throughout the SNF Project.

  6. Volatiles in inter-specific bacterial interactions

    NARCIS (Netherlands)

    Tyc, O.; Zweers, H.; De Boer, W.; Garbeva, P.V.

    2015-01-01

    The importance of volatile organic compounds for functioning of microbes is receiving increased research attention. However, to date very little is known on how inter-specific bacterial interactions effect volatiles production as most studies have been focused on volatiles produced by monocultures

  7. Autobiographical memory specificity in dissociative identity disorder

    NARCIS (Netherlands)

    Huntjens, Rafaële J. C.; Wessel, Ineke; Hermans, Dirk; van Minnen, Agnes

    2014-01-01

    A lack of adequate access to autobiographical knowledge has been related to psychopathology. More specifically, patients suffering from depression or a history of trauma have been found to be characterized by overgeneral memory, in other words, they show a relative difficulty in retrieving a specifi

  8. Familial Aggregation in Specific Language Impairment.

    Science.gov (United States)

    Tallal, Paula; Hirsch, Linda S.; Realpe-Bonilla, Teresa; Miller, Steve; Brzustowicz, Linda M.; Bartlett, Christopher; Flax, Judy F.

    2001-01-01

    A case-control family study design examined the current language-related abilities of all biological, primary relatives of probands (N=22) with specific language impairment (SLI) and of matched controls. Impairment rates for family members of SLI probands was significantly higher than for controls. Also, impairment rates estimated from a family…

  9. 31 CFR 539.311 - Specific license.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Specific license. 539.311 Section 539.311 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) OFFICE OF FOREIGN ASSETS CONTROL, DEPARTMENT OF THE TREASURY WEAPONS OF MASS DESTRUCTION TRADE CONTROL...

  10. The Domain Specificity of Academic Emotional Experiences

    Science.gov (United States)

    Goetz, Thomas; Frenzel, Anne C.; Pekrun, Reinhard; Hall, Nathan C.

    2006-01-01

    The authors analyzed the domain specificity of emotions and focused on experiences of enjoyment, anxiety, and boredom in the domains of mathematics, Latin, German, and English. On the basis of assumptions in R. Pekrun's (2000; in press) control-value theory and findings of pilot studies, the authors hypothesized the existence of a largely…

  11. Teaching Case: Enterprise Architecture Specification Case Study

    Science.gov (United States)

    Steenkamp, Annette Lerine; Alawdah, Amal; Almasri, Osama; Gai, Keke; Khattab, Nidal; Swaby, Carval; Abaas, Ramy

    2013-01-01

    A graduate course in enterprise architecture had a team project component in which a real-world business case, provided by an industry sponsor, formed the basis of the project charter and the architecture statement of work. The paper aims to share the team project experience on developing the architecture specifications based on the business case…

  12. Morphing patient-specific musculoskeletal models

    DEFF Research Database (Denmark)

    Rasmussen, John; Galibarov, Pavel E.; Al-Munajjed, Amir

    other conditions may require CT or MRI data. The method and its theoretical assumptions, advantages and limitations are presented, and several examples will illustrate morphing to patient-specific models. [1] Carbes S; Tørholm S; Rasmussen, J. A Detailed Twenty-six Segments Kinematic Foot model...

  13. Autobiographical memory specificity in dissociative identity disorder

    NARCIS (Netherlands)

    Huntjens, Rafaële J. C.; Wessel, Ineke; Hermans, Dirk; van Minnen, Agnes

    2014-01-01

    A lack of adequate access to autobiographical knowledge has been related to psychopathology. More specifically, patients suffering from depression or a history of trauma have been found to be characterized by overgeneral memory, in other words, they show a relative difficulty in retrieving a specifi

  14. Language specific bootstraps for UG categories

    NARCIS (Netherlands)

    van Kampen, N.J.

    2005-01-01

    This paper argues that the universal categories N/V are not applied to content words before the grammatical markings for reference D(eterminers) and predication I(nflection) have been acquired (van Kampen, 1997, contra Pinker, 1984). Child grammar starts as proto-grammar with language-specific opera

  15. Dancing in Place: Site-Specific Work

    Science.gov (United States)

    Metal-Corbin, Josie

    2012-01-01

    In her lecture the 2012 NDA Scholar/Artist, Josie Metal-Corbin, chronicles four decades of working with artists, educators, librarians, and scientists. The kinetic language of dance and the visual impact of specific environments provide provocative opportunities for collaboration, wherein the site becomes the framework or map for the dance design.…

  16. Understanding Bacteriophage Specificity in Natural Microbial Communities

    Directory of Open Access Journals (Sweden)

    Britt Koskella

    2013-03-01

    Full Text Available Studying the coevolutionary dynamics between bacteria and the bacteriophage viruses that infect them is critical to understanding both microbial diversity and ecosystem functioning. Phages can play a key role in shaping bacterial population dynamics and can significantly alter both intra- and inter-specific competition among bacterial hosts. Predicting how phages might influence community stability and apparent competition, however, requires an understanding of how bacteria-phage interaction networks evolve as a function of host diversity and community dynamics. Here, we first review the progress that has been made in understanding phage specificity, including the use of experimental evolution, we then introduce a new dataset on natural bacteriophages collected from the phyllosphere of horse chestnut trees, and finally we highlight that bacterial sensitivity to phage is rarely a binary trait and that this variation should be taken into account and reported. We emphasize that there is currently insufficient evidence to make broad generalizations about phage host range in natural populations, the limits of phage adaptation to novel hosts, or the implications of phage specificity in shaping microbial communities. However, the combination of experimental and genomic approaches with the study of natural communities will allow new insight to the evolution and impact of phage specificity within complex bacterial communities.

  17. Sedentary behavior and residual-specific mortality

    Science.gov (United States)

    Loprinzi, Paul D.; Edwards, Meghan K.; Sng, Eveleen; Addoh, Ovuokerie

    2016-01-01

    Background: The purpose of this study was to examine the association of accelerometer-assessed sedentary behavior and residual-specific mortality. Methods: Data from the 2003-2006 National Health and Nutrition Examination Survey (NHANES) were used (N = 5536), with follow-up through 2011. Sedentary behavior was objectively measured over 7 days via accelerometry. Results: When expressing sedentary behavior as a 60 min/day increase, the hazard ratio across the models ranged from 1.07-1.40 (P < 0.05). There was evidence of an interaction effect between sedentary behavior and total physical activity on residual-specific mortality (Hazard ratiointeraction [HR] = 0.9989; 95% CI: 0.9982-0.9997; P = 0.008). Conclusion: Sedentary behavior was independently associated with residual-specific mortality. However, there was evidence to suggest that residual-specific mortality risk was a function of sedentary behavior and total physical activity. These findings highlight the need for future work to not only examine the association between sedentary behavior and health independent of total physical activity, but evaluate whether there is a joint effect of these two parameters on health.

  18. A specification language for organisational performance indicators

    NARCIS (Netherlands)

    Popova, Viara; Treur, Jan

    2007-01-01

    A specification language for performance indicators and their relations and requirements is presented and illustrated for a case study in logistics. The language can be used in different forms, varying from informal, semiformal, graphical to formal. A software environment has been developed that s

  19. Specificity of Structural Assessment of Knowledge

    Science.gov (United States)

    Trumpower, David L.; Sharara, Harold; Goldsmith, Timothy E.

    2010-01-01

    This study examines the specificity of information provided by structural assessment of knowledge (SAK). SAK is a technique which uses the Pathfinder scaling algorithm to transform ratings of concept relatedness into network representations (PFnets) of individuals' knowledge. Inferences about individuals' overall domain knowledge based on the…

  20. An Undergraduate Laboratory Activity Demonstrating Bacteriophage Specificity

    Directory of Open Access Journals (Sweden)

    Mary E. Allen

    2013-02-01

    Full Text Available Bacteriophage are among the most diverse and numerous microbes inhabiting our planet. Yet many laboratory activities fail to engage students in meaningful exploration of their diversity, unique characteristics, and abundance. In this curriculum activity students use a standard plaque assay to enumerate bacteriophage particles from a natural sample and use the scientific method to address questions about host specificity and diversity. A raw primary sewage sample is enriched for bacteriophage using hosts in the family Enterobacteriaceae. Students hypothesize about host specificity and use quantitative data (serial dilution and plaque assay to test their hypotheses. Combined class data also help them answer questions about phage diversity. The exercise was field tested with a class of 47 students using pre- and posttests. For all learning outcomes posttest scores were higher than pretest scores at or below p = 0.01. Average individualized learning gain (G was also calculated for each learning outcome. Students’ use of scientific language in reference to bacteriophage and host interaction significantly improved (p = 0.002; G = 0.50. Improved means of expression helped students construct better hypotheses on phage host specificity (G = 0.31, p = 0.01 and to explain the plaque assay method (G = 0.33, p = 0.002. At the end of the exercise students also demonstrated improved knowledge and understanding of phage specificity as related to phage therapy in humans (p < 0.001; G = 51.

  1. Brayton Isotope Power System (BIPS) facility specification

    Energy Technology Data Exchange (ETDEWEB)

    1976-05-31

    General requirements for the Brayton Isotope Power System (BIPS)/Ground Demonstration System (GDS) assembly and test facility are defined. The facility will include provisions for a complete test laboratory for GDS checkout, performance, and endurance testing, and a contamination-controlled area for assembly, fabrication, storage, and storage preparation of GDS components. Specifications, schedules, and drawings are included.

  2. Generating focused topic-specific sentiment lexicons

    NARCIS (Netherlands)

    Jijkoun, V.; de Rijke, M.; Weerkamp, W.

    2010-01-01

    We present a method for automatically generating focused and accurate topic-specific subjectivity lexicons from a general purpose polarity lexicon that allow users to pin-point subjective on-topic information in a set of relevant documents. We motivate the need for such lexicons in the field of medi

  3. Ultrahigh Specific Impulse Nuclear Thermal Propulsion

    Energy Technology Data Exchange (ETDEWEB)

    Anne Charmeau; Brandon Cunningham; Samim Anghaie

    2009-02-09

    Research on nuclear thermal propulsion systems (NTP) have been in forefront of the space nuclear power and propulsion due to their design simplicity and their promise for providing very high thrust at reasonably high specific impulse. During NERVA-ROVER program in late 1950's till early 1970's, the United States developed and ground tested about 18 NTP systems without ever deploying them into space. The NERVA-ROVER program included development and testing of NTP systems with very high thrust (~250,000 lbf) and relatively high specific impulse (~850 s). High thrust to weight ratio in NTP systems is an indicator of high acceleration that could be achieved with these systems. The specific impulse in the lowest mass propellant, hydrogen, is a function of square root of absolute temperature in the NTP thrust chamber. Therefor optimizing design performance of NTP systems would require achieving the highest possible hydrogen temperature at reasonably high thrust to weight ratio. High hydrogen exit temperature produces high specific impulse that is a diret measure of propellant usage efficiency.

  4. Copper coating specification for the RHIC arcs

    Energy Technology Data Exchange (ETDEWEB)

    Blaskiewicz, M.

    2010-12-01

    Copper coating specifications for the RHIC arcs are given. Various upgrade scenarios are considered and calculations of resistive wall losses in the arcs are used to constrain the necessary quality and surface thickness of a copper coating. We find that 10 {mu}m of high purity copper will suffice.

  5. TAL effectors specificity stems from negative discrimination.

    Directory of Open Access Journals (Sweden)

    Basile I M Wicky

    Full Text Available Transcription Activator-Like (TAL effectors are DNA-binding proteins secreted by phytopathogenic bacteria that interfere with native cellular functions by binding to plant DNA promoters. The key element of their architecture is a domain of tandem-repeats with almost identical sequences. Most of the polymorphism is located at two consecutive amino acids termed Repeat Variable Diresidue (RVD. The discovery of a direct link between the RVD composition and the targeted nucleotide allowed the design of TAL-derived DNA-binding tools with programmable specificities that revolutionized the field of genome engineering. Despite structural data, the molecular origins of this specificity as well as the recognition mechanism have remained unclear. Molecular simulations of the recent crystal structures suggest that most of the protein-DNA binding energy originates from non-specific interactions between the DNA backbone and non-variable residues, while RVDs contributions are negligible. Based on dynamical and energetic considerations we postulate that, while the first RVD residue promotes helix breaks--allowing folding of TAL as a DNA-wrapping super-helix--the second provides specificity through a negative discrimination of matches. Furthermore, we propose a simple pharmacophore-like model for the rationalization of RVD-DNA interactions and the interpretation of experimental findings concerning shared affinities and binding efficiencies. The explanatory paradigm presented herein provides a better comprehension of this elegant architecture and we hope will allow for improved designs of TAL-derived biotechnological tools.

  6. Molecular and cellular limits to somatosensory specificity

    Directory of Open Access Journals (Sweden)

    Viana Félix

    2008-04-01

    Full Text Available Abstract Animals detect environmental changes through sensory neural mechanisms that enable them to differentiate the quality, intensity and temporal characteristics of stimuli. The 'doctrine of specific nervous energies' postulates that the different sensory modalities experienced by humans result of the activation of specific nervous pathways. Identification of functional classes of sensory receptors provided scientific support to the concept that somatosensory modalities (touch, pain, temperature, kinesthesis are subserved by separate populations of sensory receptor neurons specialized in detecting innocuous and injurious stimuli of different quality (mechanical forces, temperature, chemical compounds. The identification of receptor proteins activated by different physicochemical stimuli, in particular ion channels of the Transient Receptor Potential (TRP superfamily, has put forward the concept that specificity of peripheral sensory receptor neurons is determined by their expression of a particular "molecular sensor" that confers to each functional type its selectivity to respond with a discharge of nerve impulses to stimuli of a given quality. Nonetheless, recent experimental data suggest that the various molecular sensors proposed as specific transducer molecules for stimuli of different quality are not as neatly associated with the distinct functional types of sensory receptors as originally proposed. First, many ion channel molecules initially associated to the transduction of only one particular form of energy are also activated by stimuli of different quality, implying a limited degree of specificity in their transducing capacities. Second, molecular sensors associated with a stimulus quality and hence to a sensory receptor type and ultimately to a sensory modality may be concomitantly expressed in sensory receptor neurons functionally defined as specific for another stimulus quality. Finally, activation of voltage gated channels

  7. INSPIRE Natural Risk Zones Data Specification

    Science.gov (United States)

    Harrison, M.; Thomas, F.; Tomas, R.

    2012-04-01

    INSPIRE Directive (2007/2/EC) defines Natural Risk Zones theme as: "Vulnerable areas characterised according to natural hazards (all atmospheric, hydrologic, seismic, volcanic and wildfire phenomena that, because of their location, severity, and frequency, have the potential to seriously affect society), e.g. floods, landslides and subsidence, avalanches, forest fires, earthquakes, volcanic eruptions." This specification is the work of the Natural Risk Zones thematic working group. A multinational team of experts volunteered from the community of SDICs (Spatial Data Interest Communities) and LMOs (Legally Mandated Organisations) of INSPIRE. The data specification has been compiled using reference material submitted by SDICs and LMOs and the responses to a user requirements survey. The team themselves have had to draw on their own expertise and that of their organisations and other groups to develop agreed use cases in a selection of areas pertinent to Natural Risk Zones. The scope of the Natural Risk Zones data specification is potentially very large and this presentation will demonstrate this fact. Natural Risk Zones also involves significant engagement with other thematic areas from INSPIRE. This involvement stems from the nature of hazard, exposure, vulnerability and risk. Several other thematic areas input attributes vital to understanding the nature of hazard, yet others are vital in the understanding of exposure. In working on the scope of the Natural Risk Zones theme four use cases have been created for; Floods, Landslide, Forest Fire and Earthquake. These will be used in the presentation to demonstrate the use of the data specification. The approach taken to model Natural Risk Zones is generic in its treatment of each of hazard, exposure, vulnerability and risk, with a core model, whilst allowing extensibility to be more specific where possible and required. Flood risk is significantly more precisely defined than other hazards, due in part to the

  8. Are videogame training gains specific or general?

    Science.gov (United States)

    Oei, Adam C; Patterson, Michael D

    2014-01-01

    Many recent studies using healthy adults document enhancements in perception and cognition from playing commercial action videogames (AVGs). Playing action games (e.g., Call of Duty, Medal of Honor) is associated with improved bottom-up lower-level information processing skills like visual-perceptual and attentional processes. One proposal states a general improvement in the ability to interpret and gather statistical information to predict future actions which then leads to better performance across different perceptual/attentional tasks. Another proposal claims all the tasks are separately trained in the AVGs because the AVGs and laboratory tasks contain similar demands. We review studies of action and non-AVGs to show support for the latter proposal. To explain transfer in AVGs, we argue that the perceptual and attention tasks share common demands with the trained videogames (e.g., multiple object tracking (MOT), rapid attentional switches, and peripheral vision). In non-AVGs, several studies also demonstrate specific, limited transfer. One instance of specific transfer is the specific enhancement to mental rotation after training in games with a spatial emphasis (e.g., Tetris). In contrast, the evidence for transfer is equivocal where the game and task do not share common demands (e.g., executive functioning). Thus, the "common demands" hypothesis of transfer not only characterizes transfer effects in AVGs, but also non-action games. Furthermore, such a theory provides specific predictions, which can help in the selection of games to train human cognition as well as in the design of videogames purposed for human cognitive and perceptual enhancement. Finally this hypothesis is consistent with the cognitive training literature where most post-training gains are for tasks similar to the training rather than general, non-specific improvements.

  9. Mapping UML Component Specifications to JEE Implementations

    Directory of Open Access Journals (Sweden)

    Jyhjong Lin

    2007-01-01

    Full Text Available Component-based Software Engineering (CbSE has become a well-accepted approach for developing complex software systems due to its significant advantages on composition and reuse. In practice, however, its use still requires the conjunction of a component specification method that describes how system requirements are satisfied in terms of software components. Such a component specification is then implemented in a variety of software component models (e.g., COM+, CORBA, EJB. To achieve this, a sound mapping from the specification to a designated component model is critical. In addition, for rapid advances on Internet technologies, software systems have gradually been architected as processing in distributed environments. Since a distributed environment involves often synchronous/asynchronous messages communicating among various processes, this paper focuses therefore on the mapping issue from a component specification to a component model that particularly takes into consideration of the communicating of synchronous/asynchronous messages. To illustrate, an on-line e-Learning curriculum order system is modeled for demonstrating the mapping idea. In completing the component specification, we adopt the well-known UML Components method, while in the component model we use the Enterprise Java Beans (EJB standard in that EJB is the core component model of the JEE (J2EE platform which supports well distributed operations/services. With such a practical mapping, software systems can be developed in a more effective way by specifying requirements in UML Components and implementing software components in EJB with the communicating of synchronous/asynchronous messages among various processes.

  10. Phenylthiourea specifically reduces zebrafish eye size.

    Science.gov (United States)

    Li, Zeran; Ptak, Devon; Zhang, Liyun; Walls, Elwood K; Zhong, Wenxuan; Leung, Yuk Fai

    2012-01-01

    Phenylthiourea (PTU) is commonly used for inhibiting melanization of zebrafish embryos. In this study, the standard treatment with 0.2 mM PTU was demonstrated to specifically reduce eye size in larval fish starting at three days post-fertilization. This effect is likely the result of a reduction in retinal and lens size of PTU-treated eyes and is not related to melanization inhibition. This is because the eye size of tyr, a genetic mutant of tyrosinase whose activity is inhibited in PTU treatment, was not reduced. As PTU contains a thiocarbamide group which is presented in many goitrogens, suppressing thyroid hormone production is a possible mechanism by which PTU treatment may reduce eye size. Despite the fact that thyroxine level was found to be reduced in PTU-treated larvae, thyroid hormone supplements did not rescue the eye size reduction. Instead, treating embryos with six goitrogens, including inhibitors of thyroid peroxidase (TPO) and sodium-iodide symporter (NIS), suggested an alternative possibility. Specifically, three TPO inhibitors, including those that do not possess thiocarbamide, specifically reduced eye size; whereas none of the NIS inhibitors could elicit this effect. These observations indicate that TPO inhibition rather than a general suppression of thyroid hormone synthesis is likely the underlying cause of PTU-induced eye size reduction. Furthermore, the tissue-specific effect of PTU treatment might be mediated by an eye-specific TPO expression. Compared with treatment with other tyrosinase inhibitors or bleaching to remove melanization, PTU treatment remains the most effective approach. Thus, one should use caution when interpreting results that are obtained from PTU-treated embryos.

  11. Phenylthiourea specifically reduces zebrafish eye size.

    Directory of Open Access Journals (Sweden)

    Zeran Li

    Full Text Available Phenylthiourea (PTU is commonly used for inhibiting melanization of zebrafish embryos. In this study, the standard treatment with 0.2 mM PTU was demonstrated to specifically reduce eye size in larval fish starting at three days post-fertilization. This effect is likely the result of a reduction in retinal and lens size of PTU-treated eyes and is not related to melanization inhibition. This is because the eye size of tyr, a genetic mutant of tyrosinase whose activity is inhibited in PTU treatment, was not reduced. As PTU contains a thiocarbamide group which is presented in many goitrogens, suppressing thyroid hormone production is a possible mechanism by which PTU treatment may reduce eye size. Despite the fact that thyroxine level was found to be reduced in PTU-treated larvae, thyroid hormone supplements did not rescue the eye size reduction. Instead, treating embryos with six goitrogens, including inhibitors of thyroid peroxidase (TPO and sodium-iodide symporter (NIS, suggested an alternative possibility. Specifically, three TPO inhibitors, including those that do not possess thiocarbamide, specifically reduced eye size; whereas none of the NIS inhibitors could elicit this effect. These observations indicate that TPO inhibition rather than a general suppression of thyroid hormone synthesis is likely the underlying cause of PTU-induced eye size reduction. Furthermore, the tissue-specific effect of PTU treatment might be mediated by an eye-specific TPO expression. Compared with treatment with other tyrosinase inhibitors or bleaching to remove melanization, PTU treatment remains the most effective approach. Thus, one should use caution when interpreting results that are obtained from PTU-treated embryos.

  12. Hand Specific Representations in Language Comprehension

    Directory of Open Access Journals (Sweden)

    Claire eMoody-Triantis

    2014-06-01

    Full Text Available Theories of embodied cognition argue that language comprehension involves sensory-motor re-enactments of the actions described. However, the degree of specificity of these re-enactments as well as the relationship between action and language remains a matter of debate. Here we investigate these issues by examining how hand-specific information (left or right hand is recruited in language comprehension and action execution. An fMRI study tested right-handed participants in two separate tasks that were designed to be as similar as possible to increase sensitivity of the comparison across task: an action execution go/no-go task where participants performed right or left hand actions, and a language task where participants read sentences describing the same left or right handed actions as in the execution task. We found that language-induced activity did not match the hand-specific patterns of activity found for action execution in primary somatosensory and motor cortex, but it overlapped with pre-motor and parietal regions associated with action planning. Within these pre-motor regions, both right hand actions and sentences elicited stronger activity than left hand actions and sentences - a dominant hand effect -. Importantly, both dorsal and ventral sections of the left pre-central gyrus were recruited by both tasks, suggesting different action features being recruited. These results suggest that (a language comprehension elicits motor representations that are hand-specific and akin to multimodal action plans, rather than full action re-enactments; and (b language comprehension and action execution share schematic hand-specific representations that are richer for the dominant hand, and thus linked to previous motor experience.

  13. Digital PET compliance to EARL accreditation specifications.

    Science.gov (United States)

    Koopman, Daniëlle; Groot Koerkamp, Maureen; Jager, Pieter L; Arkies, Hester; Knollema, Siert; Slump, Cornelis H; Sanches, Pedro G; van Dalen, Jorn A

    2017-12-01

    Our aim was to evaluate if a recently introduced TOF PET system with digital photon counting technology (Philips Healthcare), potentially providing an improved image quality over analogue systems, can fulfil EANM research Ltd (EARL) accreditation specifications for tumour imaging with FDG-PET/CT. We have performed a phantom study on a digital TOF PET system using a NEMA NU2-2001 image quality phantom with six fillable spheres. Phantom preparation and PET/CT acquisition were performed according to the European Association of Nuclear Medicine (EANM) guidelines. We made list-mode ordered-subsets expectation maximization (OSEM) TOF PET reconstructions, with default settings, three voxel sizes (4 × 4 × 4 mm(3), 2 × 2 × 2 mm(3) and 1 × 1 × 1 mm(3)) and with/without point spread function (PSF) modelling. On each PET dataset, mean and maximum activity concentration recovery coefficients (RCmean and RCmax) were calculated for all phantom spheres and compared to EARL accreditation specifications. The RCs of the 4 × 4 × 4 mm(3) voxel dataset without PSF modelling proved closest to EARL specifications. Next, we added a Gaussian post-smoothing filter with varying kernel widths of 1-7 mm. EARL specifications were fulfilled when using kernel widths of 2 to 4 mm. TOF PET using digital photon counting technology fulfils EARL accreditation specifications for FDG-PET/CT tumour imaging when using an OSEM reconstruction with 4 × 4 × 4 mm(3) voxels, no PSF modelling and including a Gaussian post-smoothing filter of 2 to 4 mm.

  14. Are videogame training gains specific or general?

    Directory of Open Access Journals (Sweden)

    Adam C. Oei

    2014-04-01

    Full Text Available Many recent studies using healthy adults document enhancements in perception and cognition from playing commercial action videogames. Playing action games (e.g., Call of Duty, Medal of Honor is associated with improved bottom-up lower-level information processing skills like visual-perceptual and attentional processes. One proposal states a general improvement in the ability to interpret and gather statistical information to predict future actions which then leads to better performance across different perceptual/attentional tasks. Another proposal claims all the tasks are separately trained in the action videogames because the action videogames and laboratory tasks contain similar demands. We review studies of action and non-action videogames to show support for the latter proposal. To explain transfer in action videogames, we argue that the perceptual and attention tasks share common demands with the trained videogames (e.g., multiple object tracking, rapid attentional switches, and peripheral vision. In non-action videogames, several studies also demonstrate specific, limited transfer. One instance of specific transfer is the specific enhancement to mental rotation after training in games with a spatial emphasis (e.g, Tetris. In contrast, the evidence for transfer is equivocal where the game and task do not share common demands (e.g., executive functioning. Thus, the common demands hypothesis of transfer not only characterizes transfer effects in action videogames, but also non-action games. Furthermore, such a theory provides specific predictions, which can help in the selection of games to train human cognition as well as in the design of videogames purposed for human cognitive and perceptual enhancement. Finally this hypothesis is consistent with the cognitive training literature where most post-training gains are for tasks similar to the training rather than general, non-specific improvements.

  15. URPD: a specific product primer design tool.

    Science.gov (United States)

    Chuang, Li-Yeh; Cheng, Yu-Huei; Yang, Cheng-Hong

    2012-06-19

    Polymerase chain reaction (PCR) plays an important role in molecular biology. Primer design fundamentally determines its results. Here, we present a currently available software that is not located in analyzing large sequence but used for a rather straight-forward way of visualizing the primer design process for infrequent users. URPD (yoUR Primer Design), a web-based specific product primer design tool, combines the NCBI Reference Sequences (RefSeq), UCSC In-Silico PCR, memetic algorithm (MA) and genetic algorithm (GA) primer design methods to obtain specific primer sets. A friendly user interface is accomplished by built-in parameter settings. The incorporated smooth pipeline operations effectively guide both occasional and advanced users. URPD contains an automated process, which produces feasible primer pairs that satisfy the specific needs of the experimental design with practical PCR amplifications. Visual virtual gel electrophoresis and in silico PCR provide a simulated PCR environment. The comparison of Practical gel electrophoresis comparison to virtual gel electrophoresis facilitates and verifies the PCR experiment. Wet-laboratory validation proved that the system provides feasible primers. URPD is a user-friendly tool that provides specific primer design results. The pipeline design path makes it easy to operate for beginners. URPD also provides a high throughput primer design function. Moreover, the advanced parameter settings assist sophisticated researchers in performing experiential PCR. Several novel functions, such as a nucleotide accession number template sequence input, local and global specificity estimation, primer pair redesign, user-interactive sequence scale selection, and virtual and practical PCR gel electrophoresis discrepancies have been developed and integrated into URPD. The URPD program is implemented in JAVA and freely available at http://bio.kuas.edu.tw/urpd/.

  16. SPECIFICITIES PRESENTED IN SOME BASIC AND SPECIFIC MOTOR SKILLS OF VARIABLES TO YOUNG BASKETBALL PLAYERS

    OpenAIRE

    Hazir Salihu; Dardan Dehari

    2016-01-01

    On the basis of the number of 62 entities aged 15-16 years, male gender, address space is composed of the locomotive and the space of ten tests. Factorial procedures are addressed in all basic and specific locomotives, together as results have gained 3 dimensions latency: - Factor of specific speed and accuracy; - Factor complex locomotive specific typical for game and basketball; - Factor of speed and explosive force to the upper extremity. Such a mix of factors, may be justi...

  17. Nanoparticles for Site Specific Genome Editing

    Science.gov (United States)

    McNeer, Nicole Ali

    Triplex-forming peptide nucleic acids (PNAs) can be used to coordinate the recombination of short 50-60 by "donor DNA" fragments into genomic DNA, resulting in site-specific correction of genetic mutations or the introduction of advantageous genetic modifications. Site-specific gene editing in hematopoietic stem and progenitor cells (HSPCs) could result in treatment or cure of inherited disorders of the blood such as beta-thalassemia. Gene editing in HSPCs and differentiated T cells could help combat HIV/AIDs by modifying receptors, such as CCR5, necessary for R5-tropic HIV entry. However, translation of genome modification technologies to clinical practice is limited by challenges in intracellular delivery, especially in difficult-to-transfect hematolymphoid cells. In vivo gene editing could also provide novel treatment for systemic monogenic disorders such as cystic fibrosis, an autosomal recessive disorder caused by mutations in the cystic fibrosis transmembrane receptor. Here, we have engineered biodegradable nanoparticles to deliver oligonucleotides for site-specific genome editing of disease-relevant genes in human cells, with high efficiency, low toxicity, and editing of clinically relevant cell types. We designed nanoparticles to edit the human beta-globin and CCR5 genes in hematopoietic cells. We show that poly(lactic-co-glycolic acid) (PLGA) nanoparticles can delivery PNA and donor DNA for site-specific gene modification in human hematopoietic cells in vitro and in vivo in NOD-scid IL2rgammanull mice. Nanoparticles delivered by tail vein localized to hematopoietic compartments in the spleen and bone marrow of humanized mice, resulting in modification of the beta-globin and CCR5 genes. Modification frequencies ranged from 0.005 to 20% of cells depending on the organ and cell type, without detectable toxicity. This project developed highly versatile methods for delivery of therapeutics to hematolymphoid cells and hematopoietic stem cells, and will help to

  18. Synthesizing Distributed Protocol Specifications from a UML State Machine Modeled Service Specification

    Institute of Scientific and Technical Information of China (English)

    Jehad Al Dallal; Kassem A.Saleh

    2012-01-01

    The object-oriented paradigm is widely applied in designing and implementing communication systems.Unified Modeling Language (UML) is a standard language used to model the design of object-oriented systems.A protocol state machine is a UML adopted diagram that is widely used in designing communication protocols.It has two key attractive advantages over traditional finite state machines:modeling concurrency and modeling nested hierarchical states.In a distributed communication system,each entity of the system has its own protocol that defines when and how the entity exchanges messages with other communicating entities in the system.The order of the exchanged messages must conform to the overall service specifications of the system.In object-oriented systems,both the service and the protocol specifications are modeled in UML protocol state machines.Protocol specification synthesis methods have to be applied to automatically derive the protocol specification from the service specification.Otherwise,a time-consuming process of design,analysis,and error detection and correction has to be applied iteratively until the design of the protocol becomes error-free and consistent with the service specification.Several synthesis methods are proposed in the literature for models other than UML protocol state machines,and therefore,because of the unique features of the protocol state machines,these methods are inapplicable to services modeled in UML protocol state machines.In this paper,we propose a synthesis method that automatically synthesizes the protocol specification of distributed protocol entities from the service specification,given that both types of specifications are modeled in UML protocol state machines.Our method is based on the latest UML version (UML2.3),and it is proven to synthesize protocol specifications that are syntactically and semantically correct.As an example application,the synthesis method is used to derive the protocol specification of the H.323

  19. Site-specific weed control technologies

    DEFF Research Database (Denmark)

    Christensen, S; Søgaard, H T; Kudsk, P

    2009-01-01

    Site-specific weed control technologies are defined as machinery or equipment embedded with technologies that detect weeds growing in a crop and, taking into account predefined factors such as economics, takes action to maximise the chances of successfully controlling them. In the article, we...... describe the basic parts of site specific weed control technologies, comprising of weed sensing systems, weed management models and precision weed control implements. A review of state-of-the-art technologies shows that several weed sensing systems and precision implements have been developed over the last...... two decades, though barriers prevent their breakthrough. Most important among these is the lack of a truly robust weed recognition method, owing to mutual shading among plants and limitations in the capacity of highly accurate spraying and weeding apparatuses.   Another barrier is the lack...

  20. Cause-specific mortality after stroke

    DEFF Research Database (Denmark)

    Mogensen, Ulla Brasch; Olsen, Tom Skyhøj; Andersen, Klaus Kaae

    2013-01-01

    We investigated cause-specific mortality in relation to age, sex, stroke severity, and cardiovascular risk factor profile in the Copenhagen Stroke Study cohort with 10 years of follow-up. In a Copenhagen community, all patients admitted to the hospital with stroke during 1992-1993 (n = 988) were...... registered on admission. Evaluation included stroke severity, computed tomography scan, and a cardiovascular risk profile. Cause of death within 10 years according to death certificate information was classified as stroke, heart/arterial disease, or nonvascular disease. Competing-risks analyses were...... performed by cause-specific Cox regression after multiple imputation of missing data, assuming that values were missing at random. Death was due to stroke in 310 patients (31%), to heart/arterial disease in 209 patients (21%), and to nonvascular diseases in 289 patients (29%); 180 patients were still alive...