61 research outputs found

    Evidence for Antisense Transcription Associated with MicroRNA Target mRNAs in Arabidopsis

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    Antisense transcription is a pervasive phenomenon, but its source and functional significance is largely unknown. We took an expression-based approach to explore microRNA (miRNA)-related antisense transcription by computational analyses of published whole-genome tiling microarray transcriptome and deep sequencing small RNA (smRNA) data. Statistical support for greater abundance of antisense transcription signatures and smRNAs was observed for miRNA targets than for paralogous genes with no miRNA cleavage site. Antisense smRNAs were also found associated with MIRNA genes. This suggests that miRNA-associated “transitivity” (production of small interfering RNAs through antisense transcription) is more common than previously reported. High-resolution (3 nt) custom tiling microarray transcriptome analysis was performed with probes 400 bp 5′ upstream and 3′ downstream of the miRNA cleavage sites (direction relative to the mRNA) for 22 select miRNA target genes. We hybridized RNAs labeled from the smRNA pathway mutants, including hen1-1, dcl1-7, hyl1-2, rdr6-15, and sgs3-14. Results showed that antisense transcripts associated with miRNA targets were mainly elevated in hen1-1 and sgs3-14 to a lesser extent, and somewhat reduced in dcl11-7, hyl11-2, or rdr6-15 mutants. This was corroborated by semi-quantitative reverse transcription PCR; however, a direct correlation of antisense transcript abundance in MIR164 gene knockouts was not observed. Our overall analysis reveals a more widespread role for miRNA-associated transitivity with implications for functions of antisense transcription in gene regulation. HEN1 and SGS3 may be links for miRNA target entry into different RNA processing pathways

    Observation Of Baker's Yeast Strains Used In Biotransformation By Atomic Force Microscopy.

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    Different strains of baker's yeast (Saccharomyces cerevisiae) were imaged with an atomic force microscope (AFM). The images of uncoated and nonfixed samples were reproducible with high-constrast and nanometer-resolution. Molecules from the polysaccharide surface of the cell wall were pictured and the distance of atoms was measured. The preparation of samples was easy, suggesting that AFM is a useful tool in this type of analyses.59135-4

    Comparative study of laser and LED systems of low intensity applied to tendon healing

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    The aim of this study was to compare the effects of Low-intensity Laser Therapy (LILT) and Light Emitting Diode Therapy (LEDT) of low intensity on the treatment of lesioned Achilles tendon of rats. The experimental model consisted of a partial mechanical lesion on the right Achilles tendon deep portion of 90 rats. One hour after the lesion, the injured animals received applications of laser/LED (685, 830/630, 880 nm), and the same procedure was repeated at 24-h intervals, for 10 days. The healing process and deposition of collagen were evaluated based on a polarization microscopy analysis of the alignment and organization of collagen bundles, through the birefringence (optical retardation-OR). The results showed a real efficiency of treatments based on LEDT and confirmed that LILT seems to be effective on healing process. Although absence of coherence of LED light, tendon healing treatment with this feature was satisfactory and can certainly replace treatments based on laser light applications. Applications of infrared laser at 830 nm and LED 880 nm were more efficient when the aim is a good organization, aggregation, and alignment of the collagen bundles on tendon healing. However, more research is needed for a safety and more efficient determination of a protocol with LED

    The effects of laser irradiation on osteoblast and osteosarcoma cell proliferation and differentiation in vitro

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    Objective: The aim of this study was to investigate the effects of 670-nm, 780-nm, and 830-nm laser irradiation on cell proliferation of normal primary osteoblast ( MC3T3) and malignant osteosarcoma ( MG63) cell lines in vitro. Background: Some studies have shown that laser phototherapy is able to stimulate the osteogenesis of bone tissue, increasing osteoblast proliferation and accelerating fracture consolidation. It has been suggested that laser light may have a biostimulatory effect on tumor cells. However, the mechanism by which the laser acts on cells is not fully understood. Materials and Methods: Neonatal, murine, calvarial, osteoblastic, and human osteosarcoma cell lines were studied. A single laser irradiation was performed at three different wavelengths, at the energies of 0.5, 1, 5, and 10 J/cm(2). Twenty-four hours after laser irradiation, cell proliferation and alkaline phosphatase assays were assessed. Results: Osteoblast proliferation increased significantly after 830-nm laser irradiation ( at 10 J/cm(2)) but decreased after 780-nm laser irradiation ( at 1, 5, and 10 J/cm(2)). Osteosarcoma cell proliferation increased significantly after 670-nm ( at 5 J/cm(2)) and 780-nm laser irradiation ( at 1, 5, and 10 J/cm(2)), but not after 830-nm laser irradiation. Alkaline phosphatase ( ALP) activity in the osteoblast line was increased after 830-nm laser irradiation at 10 J/cm(2), whereas ALP activity in the osteosarcoma line was not altered, regardless of laser wavelength or intensity. Conclusion: Based on the conditions of this study, we conclude that each cell line responds differently to specific wavelength and dose combinations. Further investigations are required to investigate the physiological mechanisms responsible for the contrasting outcomes obtained when using laser irradiation on cultured normal and malignant bone cells
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