262 research outputs found

    A Fluorescent Probe for Diacetyl Detection

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    A water-soluble fluorescent probe, rhodamine B hydrazide (RBH), was prepared and its properties for recognition of diacetyl were studied. The method employs the reaction of diacetyl with RBH, a colorless and non-fluorescent rhodamine B spiro form derivative to give a pink-colored fluorescent substance. In weakly acidic media, RBH reacts more selectively with diacetyl than with other carbonyls, causing a large increase in fluorescence intensity and thereby providing an easy assay for the determination of diacetyl

    Overexpression of miR-128 specifically inhibits the truncated isoform of NTRK3 and upregulates BCL2 in SH-SY5Y neuroblastoma cells

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    <p>Abstract</p> <p>Background</p> <p>Neurotrophins and their receptors are key molecules in the regulation of neuronal differentiation and survival. They mediate the survival of neurons during development and adulthood and are implicated in synaptic plasticity. The human neurotrophin-3 receptor gene <it>NTRK3 </it>yields two major isoforms, a full-length kinase-active form and a truncated non-catalytic form, which activates a specific pathway affecting membrane remodeling and cytoskeletal reorganization. The two variants present non-overlapping 3'UTRs, indicating that they might be differentially regulated at the post-transcriptional level. Here, we provide evidence that the two isoforms of <it>NTRK3 </it>are targeted by different sets of microRNAs, small non-coding RNAs that play an important regulatory role in the nervous system.</p> <p>Results</p> <p>We identify one microRNA (miR-151-3p) that represses the full-length isoform of <it>NTRK3 </it>and four microRNAs (miR-128, miR-485-3p, miR-765 and miR-768-5p) that repress the truncated isoform. In particular, we show that the overexpression of miR-128 - a brain enriched miRNA - causes morphological changes in SH-SY5Y neuroblastoma cells similar to those observed using an siRNA specifically directed against truncated <it>NTRK3</it>, as well as a significant increase in cell number. Accordingly, transcriptome analysis of cells transfected with miR-128 revealed an alteration of the expression of genes implicated in cytoskeletal organization as well as genes involved in apoptosis, cell survival and proliferation, including the anti-apoptotic factor <it>BCL2</it>.</p> <p>Conclusions</p> <p>Our results show that the regulation of <it>NTRK3 </it>by microRNAs is isoform-specific and suggest that neurotrophin-mediated processes are strongly linked to microRNA-dependent mechanisms. In addition, these findings open new perspectives for the study of the physiological role of miR-128 and its possible involvement in cell death/survival processes.</p

    Extensions of MADM (Mosaic Analysis with Double Markers) in Mice

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    Mosaic Analysis with Double Markers (MADM) is a method for generating genetically mosaic mice, in which sibling mutant and wild-type cells are labeled with different fluorescent markers. It is a powerful tool that enables analysis of gene function at the single cell level in vivo. It requires transgenic cassettes to be located between the centromere and the mutation in the gene of interest on the same chromosome. Here we compare procedures for introduction of MADM cassettes into new loci in the mouse genome, and describe new approaches for expanding the utility of MADM. We show that: 1) Targeted homologous recombination outperforms random transgenesis in generation of reliably expressed MADM cassettes, 2) MADM cassettes in new genomic loci need to be validated for biallelic and ubiquitous expression, 3) Recombination between MADM cassettes on different chromosomes can be used to study reciprocal chromosomal deletions/duplications, and 4) MADM can be modified to permit transgene expression by combining it with a binary expression system. The advances described in this study expand current, and enable new and more versatile applications of MADM

    Racial differences in the association between partner abuse and barriers to prenatal health care among asian and native Hawaiian/other Pacific Islander women

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    Objectives Prenatal health care (PNC) is associated with positive maternal and infant health outcomes. There is limited knowledge regarding Native Hawaiians/Other Pacific Islanders (NHOPI) and Asian women’s access to PNC especially among those with partner abuse (PA) experience. The objectives of this paper were to (1) describe and examine factors associated with PNC access barriers among mothers, by race; and, (2) determine the association between PA and PNC access, by race. Methods We analyzed 2004–2007 data from Hawai‘i’s Pregnancy Risk Assessment Monitoring System (n = 7,158). The outcome is ≥1 experience with a PNC access barrier. PA is experience with physical violence from a partner. Descriptive statistics, and bivariate and multivariate logistic regression analyses stratified by race were conducted. Results The respondents included 35.7% NHOPI, 37.4% Asian, 20.1% White and 6.6% Other. More than 6% experienced PA, and 25.9% reported ≥1 PNC access barrier. Experience with PA was significantly associated with NHOPI and Asians reporting ≥1 barrier to accessing PNC, but was non-significant with Whites. Conclusions Programs should address barriers to accessing PNC, and target NHOPI and Asian mothers with PA experience to reduce the healthcare disparity and improve quality of life
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