Association Study between BDNF Gene Polymorphisms and Autism by Three-Dimensional Gel-Based Microarray

Single nucleotide polymorphisms (SNPs) are important markers which can be used in association studies searching for susceptible genes of complex diseases. High-throughput methods are needed for SNP genotyping in a large number of samples. In this study, we applied polyacrylamide gel-based microarray combined with dual-color hybridization for association study of four BDNF polymorphisms with autism. All the SNPs in both patients and controls could be analyzed quickly and correctly. Among four SNPs, only C270T polymorphism showed significant differences in the frequency of the allele (χ2 = 7.809, p = 0.005) and genotype (χ2 = 7.800, p = 0.020). In the haplotype association analysis, there was significant difference in global haplotype distribution between the groups (χ2 = 28.19, p = 3.44e-005). We suggest that BDNF has a possible role in the pathogenesis of autism. The study also show that the polyacrylamide gel-based microarray combined with dual-color hybridization is a rapid, simple and high-throughput method for SNPs genotyping, and can be used for association study of susceptible gene with disorders in large samples

Genotyping Performance between Saliva and Blood-Derived Genomic DNAs on the DMET Array: A Comparison

The Affymetrix Drug Metabolism Enzymes and Transporters (DMET) microarray is the first assay to offer a large representation of SNPs conferring genetic diversity across known pharmacokinetic markers. As a convenient and painless alternative to blood, saliva samples have been reported to work well for genotyping on the high density SNP arrays, but no reports to date have examined this application for saliva-derived DNA on the DMET platform. Genomic DNA extractions from saliva samples produced an ample quantity of genomic DNA for DMET arrays, however when human amplifiable DNA was measured, it was determined that a large percentage of this DNA was from bacteria or fungi. A mean of 37.3% human amplifiable DNA was determined for saliva-derived DNAs, which results in a significant decrease in the genotyping call rate (88.8%) when compared with blood-derived DNAs (99.1%). More interestingly, the percentage of human amplifiable DNA correlated with a higher genotyping call rate, and almost all samples with more than 31.3% human DNA produced a genotyping call rate of at least 96%. SNP genotyping results for saliva derived DNA (n = 39) illustrated a 98.7% concordance when compared with blood DNA. In conclusion, when compared with blood DNA and tested on the DMET array, saliva-derived DNA provided adequate genotyping quality with a significant lower number of SNP calls. Saliva-derived DNA does perform very well if it contains greater than 31.3% human amplifiable DNA

Docosahexaenoic acid (DHA)-induced lateral phase separation in model membrane bilayers and monolayers

This document only includes an excerpt of the corresponding thesis or dissertation. To request a digital scan of the full text, please contact the Ruth Lilly Medical Library's Interlibrary Loan Department ([email protected])

Expression and function of the PRL family of protein tyrosine phosphatase

The PRL family of enzymes constitutes a unique class of protein tyrosine phosphatase, consisting of three highly homologous members (PRL-1, PRL-2, and PRL-3). Family member PRL-3 is highly expressed in a number of tumor types and has recently gained much interest as a potential prognostic indicator of increased disease aggressiveness and poor clinical outcome for multiple human cancers. PRL-1 and PRL-2 are also known to promote a malignant phenotype in vitro, however, prior to the present study, little was known about their expression in human normal or tumor tissues. In addition, the biological function of all three PRL enzymes remains elusive and the underlying mechanisms by which they exert their effects are poorly understood. The current project was undertaken to expand our knowledge surrounding the normal cellular function of the PRL enzymes, the signaling pathways in which they operate, and the roles they play in the progression of human disease. We first characterized the tissue distribution and cell-type specific localization of PRL-1 and PRL-2 transcripts in a variety of normal and diseased human tissues using in situ hybridization. In normal, adult human tissues we found that PRL-1 and PRL-2 messages were almost ubiquitously expressed. Only highly specialized cell types, such as fibrocartilage cells, the taste buds of the tongue, and select neural cells displayed little to no expression of either transcript. In almost every other tissue and cell type examined, PRL-2 was expressed strongly while PRL-1 expression levels were variable. Each transcript was widely expressed in both proliferating and quiescent cells indicating that different tissues or cell types may display a unique physiological response to these genes. In support of this idea, we found alterations of PRL-1 and PRL-2 transcript levels in tumor samples to be highly tissue-type specific. PRL-1 expression was significantly increased in 100% of hepatocellular and gastric carcinomas, but significantly decreased in 100% of ovarian, 80% of breast, and 75% of lung tumors as compared to matched normal tissues from the same subjects. Likewise, PRL-2 expression was significantly higher in 100% of hepatocellular carcinomas, yet significantly lower in 54% of kidney carcinomas compared to matched normal specimens. PRL-1 expression was found to be associated with tumor grade in the prostate, ovary, and uterus, with patient gender in the bladder, and with patient age in the brain and skeletal muscle. These results suggest an important, but pleiotropic role for PRL-1 and PRL-2 in both normal tissue function and in the neoplastic process. These molecules may have a tumor promoting effect in some tissue types, but inhibit tumor formation or growth in others. To further elucidate the signaling pathways in which the PRLs operate, we focused on PRL-1 and used microarray and microRNA gene expression profiling to examine the global molecular changes that occur in response to stable PRL-1 overexpression in HEK293 cells. This analysis led to identification of several molecules not previously associated with PRL signaling, but whose expression was significantly altered by exogenous PRL-1 expression. In particular, Filamin A, RhoGDIα, and SPARC are attractive targets for novel mediators of PRL-1 function. We also found that PRL-1 has the capacity to indirectly influence the expression of target genes through regulation of microRNA levels and we provide evidence supporting previous observations suggesting that PRL-1 promotes cell proliferation, survival, migration, invasion, and metastasis by influencing multi-functional molecules, such as the Rho GTPases, that have essential roles in regulation of the cell cycle, cytoskeletal reorganization, and transcription factor function. The combined results of these studies have expanded our current understanding of the expression and function of the PRL family of enzymes as well as of the role these important signaling molecules play in the progression of human diseas

An exploratory study on the information seeking and use behaviour of Singapore-based Filipino domestic workers

This research focused on the information seeking and use behaviour of Filipino domestic workers (FDWs) in Singapore. The study was deemed important given the constant rise of FDWs deployed overseas. In Singapore, they make up around 40% of the 172,000 overseas Filipino workers (OFWs). Their behaviour in fulfilling information needs may be defined by their very limiting and highly homogeneous information sources. This assumption was based on previous studies (Caidi, Allard, & Quirke, 2010; Wang & Chen, 2012; Fisher, Durrance, & Hinton, 2004; Fisher, Marcoux, Miller, Sánchez, & Cunningham, 2004; Gibbs, 1994; Herzog Jr, Hofler, & Schlottmann, 1985; Herzog Jr & Schlottmann, 1983) that typecast low-paid and low-skilled migrants as ‘information poor.’ More often than not, they remain at the sidelines of information ecology – drawing them to their co-equals, with whom they share and trade information serendipitously. The study was also prompted by the need to analyse the FDWs’ profile vis-à-vis their usage and perceptions of information and communication technologies (ICT). Moreover, the study aimed to understand the FDWs’ problems related to their information seeking habits – especially about their usage (or non-usage) of electronic services. Central to this pursuit is a careful analysis of the e-government services offered by the embassy of the Republic of the Philippines, Singapore. The study employed both quantitative and qualitative research methods to address the abovementioned objectives. In all, there were 138 FDWs surveyed during a five-day ethnography in areas frequented by the FDWs in Singapore. Empirical data from this quantitative method were triangulated with participant observations gathered from fieldwork and extensive inputs from an official of the Philippine embassy that were elicited through an in-depth interview. Results showed that majority of the FDW-respondents are now connected online – but such does not adequately equate with information and digital literacy. Drawing them closer to the centre of information ecology requires support from the government, multi-sectors, and the FDWs themselves. The study saw the need to perpetuate participative learning even as many of the FDW-respondents claimed acquiring higher level education.Master of Science (Information Studies

Cellular Localization of PRL-1 and PRL-2 Gene Expression in Normal Adult Human Tissues

Recent evidence suggests that the PRL-1 and −2 phosphatases may be multifunctional enzymes with diverse roles in a variety of tissue and cell types. Northern blotting has previously shown widespread expression of both transcripts; however, little is known about the cell type-specific expression of either gene, especially in human tissues. Therefore, we investigated expression patterns for PRL-1 and −2 genes in multiple normal, adult human tissues using in situ hybridization. Although both transcripts were ubiquitously expressed, they exhibited strikingly different patterns of expression. PRL-2 was expressed heavily in almost every tissue and cell type examined, whereas PRL-1 expression levels varied considerably both between tissue types and between individuals. Widespread expression of PRL-1 and −2 in multiple organ systems suggests an important functional role for these enzymes in normal tissue homeostasis. In addition, the variable patterns of expression for these genes may provide distinct activities in each tissue or cell type

A project feasibility study on the establishment of a credit rating agency for small and medium enterprises (SMEs)

As the sole provider of Small and Medium Enterprises (SMEs) credit rating in the country, First SMEs Credit Link, Inc. aims to provide unbiased and relevant ratings to be used by financial institutions and SMEs alike in their credit risk assessments. As a credit information agency, to uphold credibility as a measure of quality to our services. Through a sound credit system, national development would be achieved. The results of the survey in Metro Manila bolstered our initial expectation. The agency\u27s primary and secondary services are expected to be well received by the Private Development Banks (PDBs) and SMEs respectively. Seven out of nine PDB\u27s would like to avail of the Credit Rating Report. The survey for Company Background Report, Financial Analysis and Property Appraisal Reports resulted in 73 percent, 75 percent and 83 percent level of acceptance among SMEs respectively. The results are very clear: First SMEs Credit Link, Inc. is acceptable to its target market. The process of services is based from existing procedures from the credit industry. This industry includes banks, private credit information agencies and CIBI. Functions of the employees are clearly defined. The process flow is organized in order to provide as a map for quality services. The financial aspect of the study proves that the credit information and ratings business for SMEs is a good project that is worth the while for implementation. Although the first two years of operations resulted in a loss, the agency managed to realize sizeable amounts of income in the last three years. This is expected to further grow in the future. In the future, First SMEs Credit Link, Inc. hopes to expand its target market not only in Metro Manila but also in other parts of the country. By that time, the proposed agency has already established its name in the credit information and ratings industry as a result of its continued improvement and customer satisfaction. The whole process of this feasibility study only proves one thing: that th