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Subconjunctival injection is a minimally invasive route for gene delivery to ocular tissues, but has traditionally been limited to use in the cornea. The accurate ocular distribution of virus has not, however, been previously investigated. Adenovirus is an attractive gene vector as it can deliver large genes and allow for short-term gene expression, but how safe it is when delivered via subconjunctival injection remains to be established. We have characterized the bio-distribution and safety of subconjunctivally administered adenovirus in Brown Norway rats. The bio-distribution and transgene duration of adenovirus carrying luciferase gene (Ad-Luci) at various time intervals were evaluated via bioluminescence imaging after subconjunctival injection. Adenovirus carrying a reporter gene, β-galactosidase (Ad-LacZ) or hrGFP (Ad-hrGFP) was administered subconjunctivally and the viral distribution in various ocular tissues was assessed by histological analysis and quantitative PCR (qPCR). Hepatic damage was assessed by biochemical and immunohistological analysis with TUNEL stain. Systemic immunogenicity was assessed by measuring serum level of TNF-α via ELISA, 2 hours and 14 days after administration of adenovirus. Retinal function was examined by electroretinography. Subconjunctival injection of Ad-Luci induced luciferase expression in the injected eyes within 24 hours, for at least 64 days. Histological analysis showed adenovirus distributed across anterior and posterior ocular tissues. qPCR demonstrated different amounts of adenovirus in different ocular tissues, with the highest amounts closest to the injection site Unlike the intravenous route, subconjunctivally delivered adenovirus did not elicit any detectable hepatic injury or systemic immunogenicity. Retinal function was unaffected by adenovirus irrespective of administration route. In conclusion, an adenoviral vector administered subconjunctivally can infiltrate into different ocular tissues and lead to short-term ocular transgene expression, without causing hepatic injury and immune activation. Therefore, subconjunctivally administered adenovirus may be a promising gene delivery approach for managing anterior and posterior segment eye diseases requiring short-term therapy.

Objective: To assess the sensitivity of a first-trimester combined screening test for fetal chromosomal abnormalities. Materials and Methods: From April 1999 to December 2002, 5,036 unselected women with singleton pregnancies underwent a first-trimester combined test (nuchal translucency thickness, pregnancy-associated plasma protein-A, and free β-human chorionic gonadotropin) screening for Down syndrome. They included 298 (5.9%) women aged 35 years or older and 4,738 (94.1%) women aged below 35 years. A positive result was defined as an estimated Down syndrome risk of greater than 1/270. Results: Twenty-five (0.50%) chromosomal abnormalities were identified, with a false-positive rate of 7.1% (333/4,717) in women younger than 35 years and a false-positive rate of 17.3% (51/294) in women of advanced maternal age. The 25 chromosomal abnormalities included trisomy 21 in six fetuses, trisomy 18 and Turner's syndrome in four fetuses each, Klinefelter's syndrome, a marker chromosome and pseudohermaphroditism in one fetus each, and structural rearrangements in eight, including four with balanced translocations. One fetus with trisomy 18 was not discovered at screening, but a fetal ventricular septal defect was found on ultrasound at 30 weeks' gestation. The detection rate for fetal chromosomal abnormalities was 95.2% (20/21). If a cutoff of 1 in 200 was used, the false-positive rate would be 5.6% with the same detection rate, whereas the test still yielded a detection rate of 90.5% (19/21) with a 5% false-positive rate. In 83.3% of cases with trisomy 21 and 5.7% of normal pregnancies, fetal nuchal translucency thickness measurement was above the 95th percentile, whereas in 0.7% of the study population and in 24% of those with fetal chromosomal abnormalities, the fetal nuchal translucency thickness was at least 3 mm. Conclusion: Our results indicate that the first-trimester combined test for Down syndrome is effective in identifying fetal chromosomal abnormalities. Moreover, the measurement of fetal nuchal translucency thickness at 10 to 13 weeks of gestation enables early detection of major fetal structural anomalies.

BACKGROUND: Chronic kidney disease (CKD) and hepatitis C virus (HCV) infection are closely linked and both increase patient mortality. The association of HCV and risk of developing end-stage renal disease (ESRD) has not been analyzed with competing risk model. METHOD: We enrolled a prospective cohort of 4,185 patients (mean age, 62 years; 41% female) registered in the CKD integrated care program at two affiliated hospitals of Kaohsiung Medical University in Taiwan between November 11, 2002 and May 31, 2009. With competing risk model, we analyzed the association of HCV infection, defined by seropositive of anti-HCV antibody, and hepatitis B virus (HBV) infection, defined by seropositive of HBV surface antigen, with the risk of entering ESRD. RESULTS: The prevalence of HCV infection was 7.6% and it increased with the CKD stages (trend test, P

Insulin-like growth factor, (IGF)-1, is produced mainly by the liver and plays important roles in promoting growth and regulating metabolism. Previous study reported that development of hepatocellular carcinoma (HCC) was accompanied by a significant reduction in serum IGF-1 levels. Here, we hypothesized that dysregulation of microRNAs (miRNA) in HCC can modulate IGF-1 expression post-transcriptionally.The miRNAs expression profiles in a dataset of 29 HCC patients were examined using illumina BeadArray. Specific miRNA (miR)-190b, which was significantly up-regulated in HCC tumor tissues when compared with paired non-tumor tissues, was among those predicted to interact with 3'-untranslated region (UTR) of IGF-1. In order to explore the regulatory effects of miR-190b on IGF-1 expression, luciferase reporter assay, quantitative real-time PCR, western blotting and immunofluorecence analysis were performed in HCC cells.Overexpression of miR-190b in Huh7 cells attenuated the expression of IGF-1, whereas inhibition of miR-190b resulted in up-regulation of IGF-1. Restoration of IGF-1 expression reversed miR-190b-mediated impaired insulin signaling in Huh7 cells, supporting that IGF-1 was a direct and functional target of miR-190b. Additionally, low serum IGF-1 level was associated with insulin resistance and poor overall survival in HCC patients.Increased expression of miR-190 may cause decreased IGF-1 in HCC development. Insulin resistance appears to be a part of the physiopathologic significance of decreased IGF-1 levels in HCC progression. This study provides a novel miRNA-mediated regulatory mechanism for controlling IGF-1 expression in HCC and elucidates the biological relevance of this interaction in HCC.

Prior cross-cultural research on dissonance has relied on cultural stereotypes in assuming that individuals from Western cultures are individualistic and have independent self-construals whereas individuals from Asian cultures are collectivistic and have interdependent self-construals. The current research made use of priming to avoid relying on cultural stereotypes and examined how having an independent or interdependent self-construal accounted for differences in dissonance experienced. A total of 120 participants who were Singapore citizens or permanent residents were randomly assigned to one of four conditions. Participants received either an independent or interdependent prime, and rated and ranked CDs before and after they made a choice between closely valued alternatives either for oneself or a close other. Results indicated that independently primed participants demonstrated significant dissonance when they made choices for themselves and close others whereas interdependently primed participants demonstrated significant dissonance when they made choices for close others but not for themselves. The study’s findings suggest that having either self-construal accounted for differences in dissonance experienced.

Wnt signaling contributes to the reprogramming and maintenance of cancer stem cell (CSC) states that are activated by epithelial--mesenchymal transition (EMT). However, the mechanistic relationship between EMT and the Wnt pathway in CSC is not entirely clear. Chromatin immunoprecipitation with highthroughput sequencing (ChIP-seq) indicated that EMT induces a switch from the β-catenin/E-cadherin/Sox15 complex to the β-catenin/Twist1/TCF4 complex, the latter of which then binds to CSC-related gene promoters. Tandem coimmunoprecipitation and re-ChIP experiments with epithelial-type cells further revealed that Sox15 associates with the β-catenin/E-cadherin complex, which then binds to the proximal promoter region of CASP3. Through this mechanism, Twist1 cleavage is triggered to regulate a β-catenin--elicited promotion of the CSC phenotype. During EMT, we documented that Twist1 binding to β-catenin enhanced the transcriptional activity of the β-catenin/TCF4 complex, including by binding to the proximal promoter region of ABCG2, a CSC marker. In terms of clinical application, our definition of a five-gene CSC signature (nuclear β-cateninHigh/nuclear Twist1High/E-cadherinLow/Sox15Low/CD133High) may provide a useful prognostic marker for human lung cancer. [ABSTRACT FROM AUTHOR]

Dyslipidemia is highly prevalent in patients with chronic kidney disease (CKD) and the relationship between dyslipidemia with renal outcomes in patients with moderate to advanced CKD remains controversial. Hence, our objective is to determine whether dyslipidemia is independently associated with rapid renal progression and progression to renal replacement therapy (RRT) in CKD patients. The study analyzed the association between lipid profile, RRT, and rapid renal progression (estimated glomerular filtration rate [eGFR] slope

BACKGROUND:Inherited cardiac conduction diseases (CCD) are rare but are caused by mutations in a myriad of genes. Recently, whole-exome sequencing has successfully led to the identification of causal mutations for rare monogenic Mendelian diseases. OBJECTIVE:To investigate the genetic background of a family affected by inherited CCD. METHODS AND RESULTS:We used whole-exome sequencing to study a Chinese family with multiple family members affected by CCD. Using the pedigree information, we proposed a heterozygous missense mutation (c.G695T, Gly232Val) in the lamin A/C (LMNA) gene as a candidate mutation for susceptibility to CCD in this family. The mutation is novel and is expected to affect the conformation of the coiled-coil rod domain of LMNA according to a structural model prediction. Its pathogenicity in lamina instability was further verified by expressing the mutation in a cellular model. CONCLUSIONS:Our results suggest that whole-exome sequencing is a feasible approach to identifying the candidate genes underlying inherited conduction diseases.

Four versions of tricistronic vectors expressing IgG1 light chain (LC), IgG1 heavy chain (HC), and dihydrofolate reductase (DHFR) in one transcript were designed to compare internal ribosome entry site (IRES) and furin-2A (F2A) for their influence on monoclonal antibody (mAb) expression level and quality in CHO DG44 cells. LC and HC genes are arranged as either the first or the second cistron. When using mAb quantification methods based on the detection antibodies against HC Fc region, F2A-mediated tricistronic vectors appeared to express mAb at higher levels than the IRES-mediated tricistronic vectors in both transient and stable transfections. Further analysis revealed that more than 40% of products detected in stably transfected pools generated using the two F2A-mediated tricistronic vectors were aggregates. LC and HC from the F2A stably transfected pools were not properly processed, giving rise to LC+F2A+HC or HC+F2A+LC fusion proteins, LC and HC polypeptides with F2A remnants, and incorrectly cleaved signal peptides. Both IRES-mediated tricistronic vectors express mAb with correct sizes and signal peptide cleavage. Arrangement of LC as the first cistron in the IRES-mediated tricistronic vectors exhibits increased mAb expression level, better growth, and minimized product aggregation, while arrangement of HC as first cistron results in low expression, slower growth, and high aggregation. The results obtained will be beneficial for designing vectors that enhance mAb expression level and quality in mammalian cells.

The juice extracted from Primus mume (Mei-Gin) is in the form of a highly concentrated dark black syrup. It is a common health supplement in Asia. The active component, 5-hydroxymethylfurfural (HMF), manifested a concentration-dependent effect on the inhibition of β-hexosaminidase degranulation upon antigen-sensitized RBL-2H3 cells. In order to investigate HMF (1) of P. mume extract, HMF was isolated and the HPLC profiles of 23 commercial Mei-Gin products (6 types) were qualitatively and quantitatively analyzed for HMF content. The samples included 10 Taiwanese Mei-Gin products, 3 Japanese Mei-Gin products, 5 specific-processing Mei-Gin products, 2 commercial plum drinks, 1 fresh mature plum extract, and 2 heat-processed samples. The HPLC profiles showed that the commercial products contained various amounts of HMF. Among them, the water extract of P. mume showed better yield of HMF. The yields from such products may differ due to different Mei-Gin original materials and different extraction and manufacturing procedures. [ABSTRACT FROM AUTHOR]

Focuses on linkage disequilibrium at the alcohol dehydrogenase genes, ADH2 and ADH3, loci and risk of alcoholism. Pairwise disequilibrium of the two functional polymorphisms and a third, presumably neutral, intronic polymorphism in ADH2 found in samples both of alcoholics and of controls from three Taiwanese populations; Comparison of the proportions of chromosomes with variants of the genes in alcoholics and control samples.