Effects of 1α,25 dihydroxyvitamin D3 and testosterone on miRNA and mRNA expression in LNCaP cells

<p>Abstract</p> <p>Background</p> <p>There is evidence from epidemiological and <it>in vitro </it>studies that the biological effects of testosterone (T) on cell cycle and survival are modulated by 1,25-dihydroxyvitamin D₃(1,25(OH)₂D₃) in prostate cancer. To investigate the cross talk between androgen- and vitamin D-mediated intracellular signaling pathways, the individual and combined effects of T and 1,25(OH)₂D₃on global gene expression in LNCaP prostate cancer cells were assessed.</p> <p>Results</p> <p>Stringent statistical analysis identifies a cohort of genes that lack one or both androgen response elements (AREs) or vitamin D response elements (VDREs) in their promoters, which are nevertheless differentially regulated by both steroids (either additively or synergistically). This suggests that mechanisms in addition to VDR- and AR-mediated transcription are responsible for the modulation of gene expression. Microarray analysis shows that fifteen miRNAs are also differentially regulated by 1,25(OH)₂D₃and T. Among these miR-22, miR-29ab, miR-134, miR-1207-5p and miR-371-5p are up regulated, while miR-17 and miR-20a, members of the miR-17/92 cluster are down regulated. A number of genes implicated in cell cycle progression, lipid synthesis and accumulation and calcium homeostasis are among the mRNA targets of these miRNAs. Thus, in addition to their well characterized effects on transcription, mediated by either or both cognate nuclear receptors, 1,25(OH)₂D₃and T regulate the steady state mRNA levels by modulating miRNA-mediated mRNA degradation, generating attenuation feedback loops that result in global changes in mRNA and protein levels. Changes in genes involved in calcium homeostasis may have specific clinical importance since the second messenger Ca²⁺is known to modulate various cellular processes, including cell proliferation, cell death and cell motility, which affects prostate cancer tumor progression and responsiveness to therapy.</p> <p>Conclusions</p> <p>These data indicate that these two hormones combine to drive a differentiated phenotype, and reinforce the idea that the age dependent decline in both hormones results in the de-differentiation of prostate tumor cells, which results in increased proliferation, motility and invasion common to aggressive tumors. These studies also reinforce the potential importance of miRNAs in prostate cancer progression and therapeutic outcomes.</p

Phenomenology of a-axis and b-axis charge dynamics from microwave spectroscopy of highly ordered YBa2Cu3O6.50 and YBa2Cu3O6.993

Extensive measurements of the microwave conductivity of highly pure and oxygen-ordered \YBCO single crystals have been performed as a means of exploring the intrinsic charge dynamics of a d-wave superconductor. Broadband and fixed-frequency microwave apparatus together provide a very clear picture of the electrodynamics of the superconducting condensate and its thermally excited nodal quasiparticles. The measurements reveal the existence of very long-lived excitations deep in the superconducting state, as evidenced by sharp cusp-like conductivity spectra with widths that fall well within our experimental bandwidth. We present a phenomenological model of the microwave conductivity that captures the physics of energy-dependent quasiparticle dynamics in a d-wave superconductor which, in turn, allows us to examine the scattering rate and oscillator strength of the thermally excited quasiparticles as functions of temperature. Our results are in close agreement with the Ferrell-Glover-Tinkham sum rule, giving confidence in both our experiments and the phenomenological model. Separate experiments for currents along the $\hat a$ and $\hat b$ directions of detwinned crystals allow us to isolate the role of the CuO chain layers in \YBCO, and a model is presented that incorporates both one-dimensional conduction from the chain electrons and two-dimensional transport associated with the \cuplane plane layers.Comment: 17 pages, 13 figure

Thrombospondin-1 Type 1 Repeats in a Model of Inflammatory Bowel Disease: Transcript Profile and Therapeutic Effects

Thrombospondin-1 (TSP-1) is a matricellular protein with regulatory functions in inflammation and cancer. The type 1 repeats (TSR) domains of TSP-1 have been shown to interact with a wide range of proteins that result in the anti-angiogenic and anti-tumor properties of TSP-1. To ascertain possible functions and evaluate potential therapeutic effects of TSRs in inflammatory bowel disease, we conducted clinical, histological and microarray analyses on a mouse model of induced colitis. We used dextran sulfate sodium (DSS) to induce colitis in wild-type (WT) mice for 7 days. Simultaneously, mice were injected with either saline or one form of TSP-1 derived recombinant proteins, containing either (1) the three type 1 repeats of the TSP-1 (3TSR), (2) the second type 1 repeat (TSR2), or (3) TSR2 with the RFK sequence (TSR2+RFK). Total RNA isolated from the mice colons were processed and hybridized to mouse arrays. Array data were validated by real-time qPCR and immunohistochemistry. Histological and disease indices reveal that the mice treated with the TSRs show different patterns of leukocytic infiltration and that 3TSR treatment was the most effective in decreasing inflammation in DSS-induced colitis. Transcriptional profiling revealed differentially expressed (DE) genes, with the 3TSR-treated mice showing the least deviation from the WT-water controls. In conclusion, this study shows that 3TSR treatment is effective in attenuating the inflammatory response to DSS injury. In addition, the transcriptomics work unveils novel genetic data that suggest beneficial application of the TSR domains in inflammatory bowel disease

Thrombospondin-1 in a Murine Model of Colorectal Carcinogenesis

Colorectal Cancer (CRC) is one of the late complications observed in patients suffering from inflammatory bowel diseases (IBD). Carcinogenesis is promoted by persistent chronic inflammation occurring in IBD. Understanding the mechanisms involved is essential in order to ameliorate inflammation and prevent CRC. Thrombospondin 1 (TSP-1) is a multidomain glycoprotein with important roles in angiogenesis. The effects of TSP-1 in colonic tumor formation and growth were analyzed in a model of inflammation-induced carcinogenesis. WT and TSP-1 deficient mice (TSP-1-/-) of the C57BL/6 strain received a single injection of azoxymethane (AOM) and multiple cycles of dextran sodium sulfate (DSS) to induce chronic inflammation-related cancers. Proliferation and angiogenesis were histologically analyzed in tumors. The intestinal transcriptome was also analyzed using a gene microarray approach. When the area containing tumors was compared with the entire colonic area of each mouse, the tumor burden was decreased in AOM/DSS-treated TSP-1-/- versus wild type (WT) mice. However, these lesions displayed more angiogenesis and proliferation rates when compared with the WT tumors. AOM-DSS treatment of TSP-1-/- mice resulted in significant deregulation of genes involved in transcription, canonical Wnt signaling, transport, defense response, regulation of epithelial cell proliferation and metabolism. Microarray analyses of these tumors showed down-regulation of 18 microRNAs in TSP-1-/- tumors. These results contribute new insights on the controversial role of TSP-1 in cancer and offer a better understanding of the genetics and pathogenesis of CRC

Gene expression during normal and FSHD myogenesis

<p>Abstract</p> <p>Background</p> <p>Facioscapulohumeral muscular dystrophy (FSHD) is a dominant disease linked to contraction of an array of tandem 3.3-kb repeats (D4Z4) at 4q35. Within each repeat unit is a gene, <it>DUX4</it>, that can encode a protein containing two homeodomains. A <it>DUX4 </it>transcript derived from the last repeat unit in a contracted array is associated with pathogenesis but it is unclear how.</p> <p>Methods</p> <p>Using exon-based microarrays, the expression profiles of myogenic precursor cells were determined. Both undifferentiated myoblasts and myoblasts differentiated to myotubes derived from FSHD patients and controls were studied after immunocytochemical verification of the quality of the cultures. To further our understanding of FSHD and normal myogenesis, the expression profiles obtained were compared to those of 19 non-muscle cell types analyzed by identical methods.</p> <p>Results</p> <p>Many of the ~17,000 examined genes were differentially expressed (> 2-fold, <it>p </it>< 0.01) in control myoblasts or myotubes vs. non-muscle cells (2185 and 3006, respectively) or in FSHD vs. control myoblasts or myotubes (295 and 797, respectively). Surprisingly, despite the morphologically normal differentiation of FSHD myoblasts to myotubes, most of the disease-related dysregulation was seen as dampening of normal myogenesis-specific expression changes, including in genes for muscle structure, mitochondrial function, stress responses, and signal transduction. Other classes of genes, including those encoding extracellular matrix or pro-inflammatory proteins, were upregulated in FSHD myogenic cells independent of an inverse myogenesis association. Importantly, the disease-linked <it>DUX4 </it>RNA isoform was detected by RT-PCR in FSHD myoblast and myotube preparations only at extremely low levels. Unique insights into myogenesis-specific gene expression were also obtained. For example, all four Argonaute genes involved in RNA-silencing were significantly upregulated during normal (but not FSHD) myogenesis relative to non-muscle cell types.</p> <p>Conclusions</p> <p><it>DUX4</it>'s pathogenic effect in FSHD may occur transiently at or before the stage of myoblast formation to establish a cascade of gene dysregulation. This contrasts with the current emphasis on toxic effects of experimentally upregulated <it>DUX4 </it>expression at the myoblast or myotube stages. Our model could explain why <it>DUX4</it>'s inappropriate expression was barely detectable in myoblasts and myotubes but nonetheless linked to FSHD.</p

DESCRIPTIVE ANALYSIS OF FATIGUE INDEX BETWEEN BASKETBALL HANDBALL AND VOLLEYBALL PLAYERS

The purpose of this study was to find out the fatigue index between Basketball, Handball and Volleyball Players. Fifteen male students from Bharathiar University Constitution College Arts Science Sivagiri were participant randomly sampled. The age of the participant was ranged from 19 to 25 years. The selected variables of fatigue index were assessed by using RAS test. Ensure the subject sprints maximally through the line each time. After 10 seconds, the next sprint starts from the opposite end of the 35 m track. Repeat this procedure until six sprints are completed. The obtained data were statistically analysed ANOVA was used to find out the significant difference, Tukey HSD test was used as a post-hoc test. An alpha level of 0.05 was used for all tests. The results indicate that there is significant difference between Basketball, Handball and Volleyball Players

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Not AvailableHistomorphological changes in neurosecretory cells of eyestalk (optic ganglia), brain and thoracic ganglia of Parapenaeopsis stylifera during different stages of ovarian maturation were recorded. There were five types of neurosecretory cells (NSCs) in eyestalk with size in range of 5-35 µm. They were distributed along medulla externa, medulla interna and medulla terminalis. Axonal terminals of these neurosecretory cells were found to terminate in sinus gland. Brain and thoracic ganglia possessed five types of neurosecretory cells such as giant neuron (80 µm), A (61-80 µm), B (41-60 µm), C (21-40 µm) and D (20 µm). They were arranged in several groups in different parts of brain - in anterior region B, C and D cells were observed, in posterior region giant neurons and Acells more in numberwhile A, B, C and D cells were found in lateral regions. Of the three regions in thoracic ganglia (anterior, middle and posterior), NSCs were distributed in anterior and posterior portions but were lacking in middle region. Aand B cells were noticed in anterior region followed by C and D cells. In posterior region, giant neurons and Acells were seen. Ovarian maturation in Parapenaeopsis stylifera seems to be under dual control of inhibitory action by the secretion of B and C cells of eyestalk and stimulatory influence by giant neuron (GN), Aand B cells of brain and thoracic ganglia as these NSCs of eyestalk were active in immature stage whereas cells of brain and thoracic ganglia were active in mature females. The active stage of secretion was characterized by hypertrophy, increase in number of nucleolus, staining intensity, granulation and migration of secretory material towards axon. Histochemical tests demonstrated that the neurosecretory cells of the shrimp were strongly positive to acid fuchsin, paraldehyde fuchsin but exhibited feeble reaction to Sudan black B and periodic acid-Schiff's reagent (PAS).Not Availabl