Thoracic aorta pseudoaneurysm with hemopericardium: unusual presentation of warfarin overdose

There have been few case reports which discuss a relationship between warfarin overdose and aortic pseudoaneurysm leakage. We report the case of a female receiving warfarin who presented with dsypnea. Her international normalized ratio was > 10. Chest radiograph revealed cardiomegaly, and chest computed tomography (CT) showed a bulging pouch-like lesion below the aortic arch greater than 6x6 cm in size and a fluid collection suggesting blood in the pericardium. Thoracic endovascular aneurysm repair (TEVAR) was successfully performed by a cardiovascular surgeon. Aortic pseudoaneurysm formation and leakage may be considered as a rare complication in patients receiving warfarin therapy. Further study regarding warfarin use and the incidence of pseudoaneurysm leakage is needed

Stimulation of transit-peptide release and ATP hydrolysis by a cochaperone during protein import into chloroplasts

Three components of the chloroplast protein translocon, Tic110, Hsp93 (ClpC), and Tic40, have been shown to be important for protein translocation across the inner envelope membrane into the stroma. We show the molecular interactions among these three components that facilitate processing and translocation of precursor proteins. Transit-peptide binding by Tic110 recruits Tic40 binding to Tic110, which in turn causes the release of transit peptides from Tic110, freeing the transit peptides for processing. The Tic40 C-terminal domain, which is homologous to the C terminus of cochaperones Sti1p/Hop and Hip but with no known function, stimulates adenosine triphosphate hydrolysis by Hsp93. Hsp93 dissociates from Tic40 in the presence of adenosine diphosphate, suggesting that Tic40 functions as an adenosine triphosphatase activation protein for Hsp93. Our data suggest that chloroplasts have evolved the Tic40 cochaperone to increase the efficiency of precursor processing and translocation

A Genome-Wide Association Study Identifies Susceptibility Variants for Type 2 Diabetes in Han Chinese

To investigate the underlying mechanisms of T2D pathogenesis, we looked for diabetes susceptibility genes that increase the risk of type 2 diabetes (T2D) in a Han Chinese population. A two-stage genome-wide association (GWA) study was conducted, in which 995 patients and 894 controls were genotyped using the Illumina HumanHap550-Duo BeadChip for the first genome scan stage. This was further replicated in 1,803 patients and 1,473 controls in stage 2. We found two loci not previously associated with diabetes susceptibility in and around the genes protein tyrosine phosphatase receptor type D (PTPRD) (P = 8.54×10−10; odds ratio [OR] = 1.57; 95% confidence interval [CI] = 1.36–1.82), and serine racemase (SRR) (P = 3.06×10−9; OR = 1.28; 95% CI = 1.18–1.39). We also confirmed that variants in KCNQ1 were associated with T2D risk, with the strongest signal at rs2237895 (P = 9.65×10−10; OR = 1.29, 95% CI = 1.19–1.40). By identifying two novel genetic susceptibility loci in a Han Chinese population and confirming the involvement of KCNQ1, which was previously reported to be associated with T2D in Japanese and European descent populations, our results may lead to a better understanding of differences in the molecular pathogenesis of T2D among various populations

A Copper Chaperone for Superoxide Dismutase Confers Three Types of CuZnSOD Activity in Arabidopsis thaliana

已知在酵母菌及哺乳動物中，copper chaperone for superoxide dismutase (CCS)是負責提供並將銅離子鑲嵌至超氧歧化酵素(superoxide dismutase，SOD)的蛋白質，而阿拉伯芥也有一個可能的CCS基因(At1g12520，AtCCS)。此基因之產物與其他物種的CCS具相似性，也可區分為三個不同的domains，分別是位於N端具有可抓住銅離子的ATX1-like domain (具MXCXXC metal binding motif)、胺基酸序列與CuZnSOD蛋白質有部分相似的central domain以及位於C端屬於CCS蛋白質所特有之序列，此區段亦具有可抓住銅離子的CXC motif。己知阿拉伯芥有三個CuZnSOD基因，分別為CSD1、CSD2及CSD3，其中CSD1及CSD2的活性分別位於細胞質及葉綠體中，由於CSD3蛋白質的C端具有peroxisome targeting signal (AKL)，所以被認為會被送入peroxisomes。然而，可能的AtCCS基因卻僅有一個，為瞭解此基因與三種CuZnSOD之間的關係，本研究利用一個同時失去三種CuZnSOD活性之Atccs knockout突變株為材料，將由The Arabidopsis Information Resource (TAIR) database中所預測的AtCCS cDNA編譯區段轉入此Atccs突變株，發現只有細胞質及peroxisome中的CuZnSOD活性會回復。在由Munich Information Centre for Protein Sequences (MIPS)中所預測的AtCCS cDNA編譯區段中，可轉譯出比TAIR database所預測AtCCS蛋白質的N端多了66個胺基酸，當我們將此基因轉入Atccs突變株，發現除上述兩種CuZnSOD外，葉綠體型CuZnSOD的活性也可被回復。經離體葉綠體蛋白質運移實驗證明，此N端多出來的66個胺基酸確實與AtCCS蛋白質進入葉綠體有關。實驗結果顯示單一AtCCS基因的產物可同時活化三種CuZnSOD活性。我們也將僅含有central domain及C-terminal domain的AtCCS蛋白質表現在Atccs突變株，發現轉植株之CuZnSOD活性並未被回復，此結果顯示在植物體中，ATX1-like domain對於AtCCS蛋白質的功能是必要的區段。 此外，我們也將水稻中可能的CCS基因(OsCCS)之cDNA對應至mature peptide的編譯區段，轉殖到Atccs突變株進行功能性互補試驗(functional complementation assay)，發現細胞質及peroxisome中的CuZnSOD活性可被回復，顯示此OsCCS基因確實具有CCS的功能，驗證了除了酵母菌的lys7突變株外，Atccs突變株也是一個適合用來探討其他植物CCS homologues功能的系統。The copper chaperone for superoxide dismutase (CCS) has been identified as a key factor integrating copper into copper/zinc superoxide dismutase (CuZnSOD) in Saccharomyces cerevisiae and mammals. In Arabidopsis thaliana, only one putative CCS gene (AtCCS, At1g12520) has been identified. The predicted AtCCS polypeptide contains three distinct domains, a central domain flanked by an ATX1-like and a C-terminal domains. The ATX1-like and the C-terminal domains contain putative copper-binding motifs. We have investigated the function of this putative AtCCS gene and shown that a cDNA encoding the open reading frame predicted by the Arabidopsis Information Resource (TAIR) complemented only the cytosolic and peroxisomal CuZnSOD activities in the Atccs knockout mutant, which has lost all CuZnSOD activities. However, a longer AtCCS cDNA, as predicted by the Munich Information Centre for Protein Sequences (MIPS), and encoding an extra 66 amino acids at the N terminus, could restore all three, including the chloroplastic, CuZnSOD activities in the Atccs mutant. The extra 66 amino acids were shown to direct the import of AtCCS into chloroplasts. Our results indicated that one AtCCS gene was responsible for the activation of all three types of CuZnSOD activity. In addition, a truncated AtCCS, containing only the central and C-terminal domains without the ATX1-like domain failed to restore any CuZnSOD activity in the Atccs mutant. This result indicates that the ATX1-like domain is essential for the copper chaperone function of AtCCS in planta. The mature peptide of a putative CCS homologue in Oryza sativa (OsCCS) was ectopically expressed in the Atccs mutant and complemented the cytosolic and peroxisomal CuZnSOD activities. The results indicate that the OsCCS protein functions as a copper chaperone donating copper ions to CuZnSOD. The Atccs mutant can serve as another system, in addition to the yeast lys7 mutant, to confirm the function of the other CCS homologues of plants.Abstract in Chinese I Abstract in English III Abbreviations V Introduction 1 Copper Ion in Living Organisms 2 Copper Trafficking Factors 2 Plant Homologues of the Copper Trafficking Factors 5 ATX1-Like Domain Study 7 Aims of the Dissertation 8 Materials and Methods 9 Plants, Growth condition, and CuSO4 Treatment 9 Analysis of the CCS Gene Sequence 9 Southern-Blot Analyses 10 Northern-Blot Analyses 10 RT-PCR and 5'-RACE of the AtCCS Gene 11 Protein Extraction and Quantification 12 Electrophoresis and SOD Activity Analyses 12 Immunoblot Analyses 12 In Vitro Transcription/Translation of AtCCScyt and AtCCScp Proteins, and Protein Import into Isolated Chloroplasts 13 Gene Construction and Plant Transformation 14 Construction and Transient Expression of the Reporter Genes 15 Results 17 AtCCS Gene in A. thaliana 17 Identification and Characterization of Superoxide Dismutase Activities in A. thaliana 18 CuZnSOD Activity and AtCCS Expression Pattern 19 Characterization of the Atccs Knockout Mutant 19 Different Recovery of CuZnSOD Activities in the Atccs Mutant Complemented with AtCCScyt, AtCCScp, and AtCCScp-mut 20 Chloroplastic Localization of AtCCScp 22 Localization of AtCCS Protein In Vivo 22 Importance of the ATX1-like Domain in Conferring CuZnSOD Activity 23 OsCCS Gene in Oryza sativa 24 Discussion 26 Figures 32 References 5

An exploratory factor analysis for developing and validating a scale of Nursing Students Competence Instrument

Background Nursing competency is a standard component of the nursing curriculum in baccalaureate nursing education in Taiwan. However, limited studies have been found on the development of a measurement for assessing Taiwanese baccalaureate-level nursing competency. Objective The aim of this study was to develop and validate a Nursing Students Competence Instrument (NSCI) for Taiwanese baccalaureate-level nursing students. Design The items of this newly developed scale were derived from a previous published qualitative study by the authors. Validity and reliability of the instruments were assessed with exploratory factor analysis. In considering external validity and homogenous characteristics, data were collected from two periods of time: February 2011 and 2012. Setting Four hundreds nursing students enrolled in the 2-year baccalaureate-level at the study university in Taiwan were invited to participate in the study. Participants Two hundreds and nine nursing students in the 2-year baccalaureate-level program were recruited. Methods Descriptive statistics and exploratory factor analysis were used to determine validity of the instrument. Cronbach alpha, split-half coefficients and item analysis verified the reliability of instrument. Results Significant levels of reliability and validity for the newly developed Nursing Student Competency Instrument were found. The competency instrument comprised four dimensions with 27 items for graduates to meet to determine their nursing competency. Four factors were analyzed and categorized as integrating care abilities, leading humanity concerns, advancing career talents, and dealing with tension, and explained 22.29%, 18.59%, 15.99% and 11.23% of total variance, respectively; these four explained 68.09% of the total variance. Conclusions Results support validation of the new nursing competence assessment scale for Taiwanese nursing students at baccalaureate levels. The authors recommend that the NSCI could be applied in the nursing schools to evaluate the learning outcomes of nursing students' competence

Targeting Chondroitin Sulfate Reduces Invasiveness of Glioma Cells by Suppressing CD44 and Integrin β1 Expression

Chondroitin sulfate (CS) is a major component of the extracellular matrix found to be abnormally accumulated in several types of cancer tissues. Previous studies have indicated that CS synthases and modification enzymes are frequently elevated in human gliomas and are associated with poor prognosis. However, the underlying mechanisms of CS in cancer progression and approaches for interrupting its functions in cancer cells remain largely unexplored. Here, we have found that CS was significantly enriched surrounding the vasculature in a subset of glioma tissues, which was akin to the perivascular niche for cancer-initiating cells. Silencing or overexpression of the major CS synthase, chondroitin sulfate synthase 1 (CHSY1), significantly regulated the glioma cell invasive phenotypes and modulated integrin expression. Furthermore, we identified CD44 as a crucial chondroitin sulfate proteoglycan (CSPG) that was modified by CHSY1 on glioma cells, and the suppression of CS formation on CD44 by silencing the CHSY1-inhibited interaction between CD44 and integrin β1 on the adhesion complex. Moreover, we tested the CS-specific binding peptide, resulting in the suppression of glioma cell mobility in a fashion similar to that observed upon the silencing of CHSY1. In addition, the peptide demonstrated significant affinity to CD44, promoted CD44 degradation, and suppressed integrin β1 expression in glioma cells. Overall, this study proposes a potential regulatory loop between CS, CD44, and integrin β1 in glioma cells, and highlights the importance of CS in CD44 stability. Furthermore, the targeting of CS by specific binding peptides has potential as a novel therapeutic strategy for glioma

A Copper Chaperone for Superoxide Dismutase That Confers Three Types of Copper/Zinc Superoxide Dismutase Activity in Arabidopsis

The copper chaperone for superoxide dismutase (CCS) has been identified as a key factor integrating copper into copper/zinc superoxide dismutase (CuZnSOD) in yeast (Saccharomyces cerevisiae) and mammals. In Arabidopsis (Arabidopsis thaliana), only one putative CCS gene (AtCCS, At1g12520) has been identified. The predicted AtCCS polypeptide contains three distinct domains: a central domain, flanked by an ATX1-like domain, and a C-terminal domain. The ATX1-like and C-terminal domains contain putative copper-binding motifs. We have investigated the function of this putative AtCCS gene and shown that a cDNA encoding the open reading frame predicted by The Arabidopsis Information Resource complemented only the cytosolic and peroxisomal CuZnSOD activities in the Atccs knockout mutant, which has lost all CuZnSOD activities. However, a longer AtCCS cDNA, as predicted by the Munich Information Centre for Protein Sequences and encoding an extra 66 amino acids at the N terminus, could restore all three, including the chloroplastic CuZnSOD activities in the Atccs mutant. The extra 66 amino acids were shown to direct the import of AtCCS into chloroplasts. Our results indicated that one AtCCS gene was responsible for the activation of all three types of CuZnSOD activity. In addition, a truncated AtCCS, containing only the central and C-terminal domains without the ATX1-like domain failed to restore any CuZnSOD activity in the Atccs mutant. This result indicates that the ATX1-like domain is essential for the copper chaperone function of AtCCS in planta