Development of a Custom-Made 3D Printing Protocol with Commercial Resins for Manufacturing Microfluidic Devices.

The combination of microfluidics and photo-polymerization techniques such as stereolithography (SLA) has emerged as a new field which has a lot of potential to influence in such important areas as biological analysis, and chemical detection among others. However, the integration between them is still at an early stage of development. In this article, after analyzing the resolution of a custom SLA 3D printer with commercial resins, microfluidic devices were manufactured using three different approaches. First, printing a mold with the objective of creating a Polydimethylsiloxane (PDMS) replica with the microfluidic channels; secondly, open channels have been printed and then assembled with a flat cover of the same resin material. Finally, a closed microfluidic device has also been produced in a single process of printing. Important results for 3D printing with commercial resins have been achieved by only printing one layer on top of the channel. All microfluidic devices have been tested successfully for pressure-driven fluid flow.

Development of a novel splice array platform and its application in the identification of alternative splice variants in lung cancer.

BACKGROUND: Microarrays strategies, which allow for the characterization of thousands of alternative splice forms in a single test, can be applied to identify differential alternative splicing events. In this study, a novel splice array approach was developed, including the design of a high-density oligonucleotide array, a labeling procedure, and an algorithm to identify splice events. RESULTS: The array consisted of exon probes and thermodynamically balanced junction probes. Suboptimal probes were tagged and considered in the final analysis. An unbiased labeling protocol was developed using random primers. The algorithm used to distinguish changes in expression from changes in splicing was calibrated using internal non-spliced control sequences. The performance of this splice array was validated with artificial constructs for CDC6, VEGF, and PCBP4 isoforms. The platform was then applied to the analysis of differential splice forms in lung cancer samples compared to matched normal lung tissue. Overexpression of splice isoforms was identified for genes encoding CEACAM1, FHL-1, MLPH, and SUSD2. None of these splicing isoforms had been previously associated with lung cancer. CONCLUSIONS: This methodology enables the detection of alternative splicing events in complex biological samples, providing a powerful tool to identify novel diagnostic and prognostic biomarkers for cancer and other pathologies.

Genomic resources for a commercial flatfish, the Senegalese sole (Solea senegalensis): EST sequencing, oligo microarray design, and development of the Soleamold bioinformatic platform.

BACKGROUND: The Senegalese sole, Solea senegalensis, is a highly prized flatfish of growing commercial interest for aquaculture in Southern Europe. However, despite the industrial production of Senegalese sole being hampered primarily by lack of information on the physiological mechanisms involved in reproduction, growth and immunity, very limited genomic information is available on this species. RESULTS: Sequencing of a S. senegalensis multi-tissue normalized cDNA library, from adult tissues (brain, stomach, intestine, liver, ovary, and testis), larval stages (pre-metamorphosis, metamorphosis), juvenile stages (post-metamorphosis, abnormal fish), and undifferentiated gonads, generated 10,185 expressed sequence tags (ESTs). Clones were sequenced from the 3'-end to identify isoform specific sequences. Assembly of the entire EST collection into contigs gave 5,208 unique sequences of which 1,769 (34%) had matches in GenBank, thus showing a low level of redundancy. The sequence of the 5,208 unigenes was used to design and validate an oligonucleotide microarray representing 5,087 unique Senegalese sole transcripts. Finally, a novel interactive bioinformatic platform, Soleamold, was developed for the Senegalese sole EST collection as well as microarray and ISH data. CONCLUSION: New genomic resources have been developed for S. senegalensis, an economically important fish in aquaculture, which include a collection of expressed genes, an oligonucleotide microarray, and a publicly available bioinformatic platform that can be used to study gene expression in this species. These resources will help elucidate transcriptional regulation in wild and captive Senegalese sole for optimization of its production under intensive culture conditions.

Plant polyphenol intake alters gene expression in canine leukocytes.

BACKGROUND/AIMS: Polyphenol compounds may explain most of the health-related beneficial effects of plants and vegetables, mainly through their antioxidant properties. The aim of the study was to assess the main changes on leukocyte gene expression of dogs caused by intake of three natural polyphenol-rich extracts and to compare them with caloric restriction. METHODS: 20 female dogs were divided into 5 groups: control fed ad libitum (C), caloric-restricted to 30% less than control (CR), and 3 groups fed ad libitum supplemented with citrus extract (CE), green tea extract (GTE) or grape seed extract (GSE). Leukocytes gene expression was analyzed in a specially designed microarray. RESULTS: CE treatment mainly downregulated genes related to inflammative (IL-8, VLA-4) and cytotoxic response (GRP 58) as well as proliferation of leukocytes. GTE induced gene expression related to leukocyte proliferation and signaling (GNAQ, PKC-B). GSE upregulated some of the genes increased by CE treatment. CR downregulated genes related with energy metabolism (ATP5A1, COX7C) and inflammatory markers (VLA-4). CONCLUSION: A chronic ingestion of citric, grape seed and green tea polyphenols is able to modulate canine leukocyte functions through changes in gene expression. CE ingestion reduces expression of some genes also diminished by a 30% caloric restriction.