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Background: Many quality-of-life studies have been conducted in healthcare settings, but few have used Microsoft Excel to incorporate Cronbach's α with dimension coefficient (DC) for describing a scale's characteristics. To present a computer module that can report a scale's validity, we manipulated datasets to verify a DC that can be used as a factor retention criterion for demonstrating its usefulness in a patient safety culture survey (PSC).Methods: Microsoft Excel Visual Basic for Applications was used to design a computer module for simulating 2000 datasets fitting the Rasch rating scale model. The datasets consisted of (i) five dual correlation coefficients (correl. = 0.3, 0.5, 0.7, 0.9, and 1.0) on two latent traits (i.e., true scores) following a normal distribution and responses to their respective 1/3 and 2/3 items in length; (ii) 20 scenarios of item lengths from 5 to 100; and (iii) 20 sample sizes from 50 to 1000. Each item containing 5-point polytomous responses was uniformly distributed in difficulty across a ± 2 logit range. Three methods (i.e., dimension interrelation ≥0.7, Horn's parallel analysis (PA) 95% confidence interval, and individual random eigenvalues) were used for determining one factor to retain. DC refers to the binary classification (1 as one factor and 0 as many factors) used for examining accuracy with the indicators sensitivity, specificity, and area under receiver operating characteristic curve (AUC). The scale's reliability and DC were simultaneously calculated for each simulative dataset. PSC real data were demonstrated with DC to interpret reports of the unit-based construct validity using the author-made MS Excel module.Results: The DC method presented accurate sensitivity (=0.96), specificity (=0.92) with a DC criterion (≥0.70), and AUC (=0.98) that were higher than those of the two PA methods. PA combined with DC yielded good sensitivity (=0.96), specificity (=1.0) with a DC criterion (≥0.70), and AUC (=0.99).Conclusions: Advances in computer technology may enable healthcare users familiar with MS Excel to apply DC as a factor retention criterion for determining a scale's unidimensionality and evaluating a scale's quality. [ABSTRACT FROM AUTHOR]

Background: Many continuous item responses (CIRs) are encountered in healthcare settings, but no one uses item response theory's (IRT) probabilistic modeling to present graphical presentations for interpreting CIR results. A computer module that is programmed to deal with CIRs is required. To present a computer module, validate it, and verify its usefulness in dealing with CIR data, and then to apply the model to real healthcare data in order to show how the CIR that can be applied to healthcare settings with an example regarding a safety attitude survey.Methods: Using Microsoft Excel VBA (Visual Basic for Applications), we designed a computer module that minimizes the residuals and calculates model's expected scores according to person responses across items. Rasch models based on a Wright map and on KIDMAP were demonstrated to interpret results of the safety attitude survey.Results: The author-made CIR module yielded OUTFIT mean square (MNSQ) and person measures equivalent to those yielded by professional Rasch Winsteps software. The probabilistic modeling of the CIR module provides messages that are much more valuable to users and show the CIR advantage over classic test theory.Conclusions: Because of advances in computer technology, healthcare users who are familiar to MS Excel can easily apply the study CIR module to deal with continuous variables to benefit comparisons of data with a logistic distribution and model fit statistics. [ABSTRACT FROM AUTHOR]

Background: Highly desaturated n-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are synthesized by desaturases and elongase. They exert hepatoprotective effects to prevent alcoholic fatty liver syndrome or cholestatic liver injury. However, it is unclear how n-3 PUFAs improve immune function in liver. Vibrio vulnificus, a gram-negative bacterial pathogen, causes high mortality of aquaculture fishes upon infection. Humans can become infected with V. vulnificus through open wounds or by eating raw seafood, and such infections may result in systemic septicemia. Moreover, patients with liver diseases are vulnerable to infection, and are more likely than healthy persons to present with liver inflammation following infection. This study quantified n-3 PUFAs and their anti-bacterial effects in Fadsd6 and Elvol5a transgenic zebrafish. Results: Two transgenic zebrafish strains with strong liver specific expression of Fadsd6 and Elvol5a (driven by the zebrafish Fabp10 promoter) were established using the Tol2 system. Synthesis of n-3 PUFAs in these strains were increased by 2.5-fold as compared to wild type (Wt) fish. The survival rate in 24 h following challenge with V. vulnificus was 20 % in Wt, but 70 % in the transgenic strains. In addition, the bacteria counts in transgenic fish strains were significantly decreased. The expression levels of pro-inflammatory genes, such as TNF- α, IL-1β, and NF-κB, were suppressed between 9 and 12 h after challenge. This study confirms the anti-bacterial function of n-3 PUFAs in a transgenic zebrafish model. Conclusions: Fadsd6 and Elvol5a transgenic zebrafish are more resistant to V. vulnificus infection, and enhance survival by diminishing the attendant inflammatory response. [ABSTRACT FROM AUTHOR]

Background: Metabolic variations exist between the methionine salvage pathway of humans and a number of plants and microbial pathogens. 5-Methylthioribose (MTR) kinase is a key enzyme required for methionine salvage in plants and many bacteria. The absence of a mammalian homolog suggests that MTR kinase is a good target for the design of specific herbicides or antibiotics. Results: The structure of Arabidopsis thaliana MTR kinase co-crystallized with ATPγS and MTR has been determined at 1.9 Å resolution. The structure is similar to B. subtilis MTR kinase and has the same protein kinase fold observed in other evolutionarily related protein kinase-like phosphotransferases. The active site is comparable between the two enzymes with the DXE-motif coordinating the nucleotide-Mg, the D238 of the HGD catalytic loop polarizing the MTR O1 oxygen, and the RR-motif interacting with the substrate MTR. Unlike its bacterial homolog, however, the Gly-rich loop (G-loop) of A. thaliana MTR kinase has an extended conformation, which shields most of the active site from solvent, a feature that resembles eukaryotic protein kinases more than the bacterial enzyme. The G- and W-loops of A. thaliana and B. subtilis MTR kinase adopt different conformations despite high sequence similarity. The ATPγS analog was hydrolyzed during the co-crystallization procedure, resulting in ADP in the active site. This suggests that the A. thaliana enzyme, like its bacterial homolog, may have significant ATPase activity in the absence of MTR. Conclusion: The structure of A. thaliana MTR kinase provides a template for structure-based design of agrochemicals, particularly herbicides whose effectiveness could be regulated by nutrient levels. Features of the MTR binding site offer an opportunity for a simple organic salt of an MTR analog to specifically inhibit MTR kinase. [ABSTRACT FROM AUTHOR]

Background: The meniscus plays a vital role in the normal biomechanics of the knee. However, it is not well studied at the molecular level. The purpose of this study was to determine whether molecular and pathological changes in the meniscal tissue vary depending on the presence or absence of meniscal and/or anterior cruciate ligament tear (ACL)., Methods: Six normal menisci (group A), seven simple torn menisci (group B) and seven torn menisci with concomitant anterior cruciate ligament tears (group C) were collected. We observed the pathological changes in the menisci and used real-time polymerase chain reaction along with immunohistochemistry and in situ hybridisation to examine the levels of ACAN, ADAMTS5, COL10A1, CEBPβ, MMP13 and miR-381-3p, miR-455-3p, miR-193b-3p, miR-92a-3p, respectively. Patients were scored preoperatively and postoperatively using the Lysholm Knee Scoring Scale and International Knee Documentation Committee Subjective Knee Evaluation Form., Results: Compared with group A, the expression levels of ADAMTS5, COL10A1, CEBPβ, and MMP13 and all the miRNAs were increased while ACAN was down-regulated in groups B and C. Additionally, the gene expression and miRNA levels were higher in group C than that in group B, except for ACAN, which was lower. Several fibrochondrocytes strongly expressed ADAMTS5, CEBPβ, and MMP13 in groups B and C and had high levels of miR-381-3p and miR-455-3p than that in group A. Postoperative Lysholm and IKDC scores were higher in group B than in group C., Conclusions: Our findings suggest that the meniscus tended to degenerate after it was injured, especially when combined with a torn ACL. The miRNAs investigated in this study might also contribute to meniscus degeneration. Patients with a combined injury patterns might have relatively worse joint function.

Background: Many quality-of-life studies have been conducted in healthcare settings, but few have used Microsoft Excel to incorporate Cronbach's α with dimension coefficient (DC) for describing a scale's characteristics. To present a computer module that can report a scale's validity, we manipulated datasets to verify a DC that can be used as a factor retention criterion for demonstrating its usefulness in a patient safety culture survey (PSC)., Methods: Microsoft Excel Visual Basic for Applications was used to design a computer module for simulating 2000 datasets fitting the Rasch rating scale model. The datasets consisted of (i) five dual correlation coefficients (correl. = 0.3, 0.5, 0.7, 0.9, and 1.0) on two latent traits (i.e., true scores) following a normal distribution and responses to their respective 1/3 and 2/3 items in length; (ii) 20 scenarios of item lengths from 5 to 100; and (iii) 20 sample sizes from 50 to 1000. Each item containing 5-point polytomous responses was uniformly distributed in difficulty across a ± 2 logit range. Three methods (i.e., dimension interrelation ≥0.7, Horn's parallel analysis (PA) 95% confidence interval, and individual random eigenvalues) were used for determining one factor to retain. DC refers to the binary classification (1 as one factor and 0 as many factors) used for examining accuracy with the indicators sensitivity, specificity, and area under receiver operating characteristic curve (AUC). The scale's reliability and DC were simultaneously calculated for each simulative dataset. PSC real data were demonstrated with DC to interpret reports of the unit-based construct validity using the author-made MS Excel module., Results: The DC method presented accurate sensitivity (=0.96), specificity (=0.92) with a DC criterion (≥0.70), and AUC (=0.98) that were higher than those of the two PA methods. PA combined with DC yielded good sensitivity (=0.96), specificity (=1.0) with a DC criterion (≥0.70), and AUC (=0.99)., Conclusions: Advances in computer technology may enable healthcare users familiar with MS Excel to apply DC as a factor retention criterion for determining a scale's unidimensionality and evaluating a scale's quality.

Background: Many continuous item responses (CIRs) are encountered in healthcare settings, but no one uses item response theory's (IRT) probabilistic modeling to present graphical presentations for interpreting CIR results. A computer module that is programmed to deal with CIRs is required. To present a computer module, validate it, and verify its usefulness in dealing with CIR data, and then to apply the model to real healthcare data in order to show how the CIR that can be applied to healthcare settings with an example regarding a safety attitude survey., Methods: Using Microsoft Excel VBA (Visual Basic for Applications), we designed a computer module that minimizes the residuals and calculates model's expected scores according to person responses across items. Rasch models based on a Wright map and on KIDMAP were demonstrated to interpret results of the safety attitude survey., Results: The author-made CIR module yielded OUTFIT mean square (MNSQ) and person measures equivalent to those yielded by professional Rasch Winsteps software. The probabilistic modeling of the CIR module provides messages that are much more valuable to users and show the CIR advantage over classic test theory., Conclusions: Because of advances in computer technology, healthcare users who are familiar to MS Excel can easily apply the study CIR module to deal with continuous variables to benefit comparisons of data with a logistic distribution and model fit statistics.

Background: Highly desaturated n-3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are synthesized by desaturases and elongase. They exert hepatoprotective effects to prevent alcoholic fatty liver syndrome or cholestatic liver injury. However, it is unclear how n-3 PUFAs improve immune function in liver. Vibrio vulnificus, a gram-negative bacterial pathogen, causes high mortality of aquaculture fishes upon infection. Humans can become infected with V. vulnificus through open wounds or by eating raw seafood, and such infections may result in systemic septicemia. Moreover, patients with liver diseases are vulnerable to infection, and are more likely than healthy persons to present with liver inflammation following infection. This study quantified n-3 PUFAs and their anti-bacterial effects in Fadsd6 and Elvol5a transgenic zebrafish., Results: Two transgenic zebrafish strains with strong liver specific expression of Fadsd6 and Elvol5a (driven by the zebrafish Fabp10 promoter) were established using the Tol2 system. Synthesis of n-3 PUFAs in these strains were increased by 2.5-fold as compared to wild type (Wt) fish. The survival rate in 24 h following challenge with V. vulnificus was 20 % in Wt, but 70 % in the transgenic strains. In addition, the bacteria counts in transgenic fish strains were significantly decreased. The expression levels of pro-inflammatory genes, such as TNF-α, IL-1β, and NF-κB, were suppressed between 9 and 12 h after challenge. This study confirms the anti-bacterial function of n-3 PUFAs in a transgenic zebrafish model., Conclusions: Fadsd6 and Elvol5a transgenic zebrafish are more resistant to V. vulnificus infection, and enhance survival by diminishing the attendant inflammatory response.

Background: The discovery of cancer stem cells and tumor heterogeneity prompted the exploration of additional mechanisms aside from genetic mutations for carcinogenesis and cancer progression. The aim of the present study was to investigate the effect of cell fusion between mesenchymal stem cells and the gastric epithelial cells in tumorigenesis., Methods: Cell fusion between cord blood mesenchymal stem cells and human gastric epithelial cells was performed in vitro. Cell scratch and transwell assays were performed to determine migration and invasion abilities of the hybrids. The expressions of epithelial-mesenchymal transition-related proteins and genes were analyzed by immunocytochemistry and real time quantitative PCR. Tumorigenesis of the hybrids was evaluated through in vivo inoculation in nude mice., Results: Hybrids expressed the phenotypes of both donor cells. Aneuploidy was observed in 84.1% of cells. The hybrids showed increased proliferation, migration and invasion abilities compared with the parental cells. In addition, the expression of N-cadherin and vimentin in the hybrids was significantly higher than that of the epithelial cells, and the mRNA expression of the epithelial-mesenchymal transition-related genes, Twist and Slug, in the hybrids was also increased compared with that of the parental epithelial cells. Furthermore, the hybrids formed masses of epithelial origin with glandular structures in BALB/c nude mice., Conclusions: These findings suggest that cell fusion between gastric epithelial cells and mesenchymal stem cells may result in epithelial to mesenchymal transition and malignant transformation.

Background: Purifying stem cells is an inevitable process for further investigation and cell-therapy. Sorting side population (SP) cells is generally regarded as an effective method to enrich for progenitor cells. This study was to explore whether sorting SP could enrich for the Musashi1 (Msi1) positive cells from Msi1 high expression cells (Msi1(high) cells) derived from mouse embryonic stem cells (ESCs) in vitro., Results: In this study, Msi1(high) cell population derived from ESCs were stained by Hoechst 33342, and then the SP and non-SP (NSP) fractions were analyzed and sorted by fluorescence activated cell sorter. Subsequently, the expressions of Msi1 and other markers for neural and intestinal stem cells in SP and NSP were respectively detected. SP and NSP cells were hypodermically engrafted into the backs of NOD/SCID mice to form grafts. The developments of neural and intestinal epithelial cells in these grafts were investigated. SP fraction was identified and isolated from Msi1(high) cell population. The expression of Msi1 in SP fraction was significantly higher than that in NSP fraction and unsorted Msi1(high) cells (P< 0.05). Furthermore, the markers for neural cells and intestinal epithelial cells were more highly expressed in the grafts from SP fraction than those from NSP fraction (P< 0.05)., Conclusions: SP fraction, isolated from Msi1(high) cells, contains almost all the Msi1-positive cells and has the potential to differentiate into neural and intestinal epithelial cells in vivo. Sorting SP fraction could be a convenient and practical method to enrich for Msi1-positive cells from the differentiated cell population derived from ESCs.

Background: Metabolic variations exist between the methionine salvage pathway of humans and a number of plants and microbial pathogens. 5-Methylthioribose (MTR) kinase is a key enzyme required for methionine salvage in plants and many bacteria. The absence of a mammalian homolog suggests that MTR kinase is a good target for the design of specific herbicides or antibiotics., Results: The structure of Arabidopsis thaliana MTR kinase co-crystallized with ATPgammaS and MTR has been determined at 1.9 A resolution. The structure is similar to B. subtilis MTR kinase and has the same protein kinase fold observed in other evolutionarily related protein kinase-like phosphotransferases. The active site is comparable between the two enzymes with the DXE-motif coordinating the nucleotide-Mg, the D238 of the HGD catalytic loop polarizing the MTR O1 oxygen, and the RR-motif interacting with the substrate MTR. Unlike its bacterial homolog, however, the Gly-rich loop (G-loop) of A. thaliana MTR kinase has an extended conformation, which shields most of the active site from solvent, a feature that resembles eukaryotic protein kinases more than the bacterial enzyme. The G- and W-loops of A. thaliana and B. subtilis MTR kinase adopt different conformations despite high sequence similarity. The ATPgammaS analog was hydrolyzed during the co-crystallization procedure, resulting in ADP in the active site. This suggests that the A. thaliana enzyme, like its bacterial homolog, may have significant ATPase activity in the absence of MTR., Conclusion: The structure of A. thaliana MTR kinase provides a template for structure-based design of agrochemicals, particularly herbicides whose effectiveness could be regulated by nutrient levels. Features of the MTR binding site offer an opportunity for a simple organic salt of an MTR analog to specifically inhibit MTR kinase.