Relationships between renal cytoplasmic and nuclear aldosterone-receptors

Relationships between renal cytoplasmic and nuclear aldosteronereceptors.Three 3H-aldosterone receptor complexes have been recovered from rat kidneys: 1) cytosol (high speed supernatants), 2) Tris-soluble nuclear (obtained by an osmotic shock procedure), and 3) chromatin-bound (prepared by extracting post-shock nuclei with 0.4 M KCl).Glycerol density gradient analyses of cytosol labelled in vivo or in vitro with 3H-aldosterone yielded two specific peaks -4.5S and 8.5S.These peaks were sensitive to salt concentration; 0.4 M KCl shifted the 8.5S to 4.5S and the addition of Ca++ (6 mM) resulted in a further shift to 3.5S.The Tris-soluble nuclear species sedimented at 3S and the chromatin-bound species at 4S.The time-course of generation of the 3H-aldosterone-labelled cytosol and nuclear receptor species was studied in vivo and in vitro by tissue slice and reconstitution methods.The results obtained are consistent with a three-step mechanism: cytosol (8.5S or 4.5S)→ Tris-soluble nuclear (3S)→ chromatin-bound (4S).Alternatively, the 3S and 4S complexes may be attached to independent nuclear sites.The formation of the chromatin-bound species was temperature sensitive and failed to form at 0°C.Pre-treatment with DNase but not RNase impaired the generation of both the Tris-soluble nuclear and chromatin-bound species.These results imply a close association between nuclear aldosterone-receptor complexes and intact DNA

Anti-oxidant NAC attenuated the decrease in fitness caused by FLT.

<p>Fitness was measured by the number of sigmoidal body bends per worm per minute. FLT concentration = 200μM. Anti-oxidant concentration = 100μM. Statistics were calculated with a two way ANOVA with Turkey’s multiple comparisons test from 10 worms per treatment condition over ≥3 independent trials. FLT compared to control of that same time point. FLT + NAC compared to FLT of that same time point. * = P<0.05, ** = P<0.01, *** = P<0.001, n.s. = not significant.</p

Overlap of thymidine analogue induced DEGs with genes known to be regulated by HIF-1, CEP-1 and SKN-1 after 72h exposure.

<p>The expression direction (Up or Down) of thymidine analogue DEGs, with an adjusted P-value <0.01, at 72h, are compared to respectively Up or Down regulated genes from the different conditions taken from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187424#pone.0187424.ref044" target="_blank">44</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187424#pone.0187424.ref046" target="_blank">46</a>]. The numbers of genes that overlap per condition at each time point are given with their representative percentage of thymidine analogue regulated genes between brackets. Over-represented P-value: * = P<0.05, ** = P<0.01, *** = P<0.001, n.s. = not significant. n.a. = not applicable.</p

NRTIs caused average lifespan extension.

<p>Lifespan extension is dependent on the timing of exposure: A, exposure from larval L1; B, exposure from larval L4. NRTI concentration = 200μM. All animals are N2 at 20°C, mean and maximum refer to the amount of days after L1 (A), and L4 (B). Statistics of NRTIs at L4 were conducted compared their respective DMSO control (AZT, d4T, ddI, & FLT = 0.067%; ddC = 0.2%). All other statistics are compared to controls. SEM = standard error of the mean.</p

Clinical characteristics of patients whose fibrovascular membranes where used for mRNA analysis.

<p>Clinical characteristics of patients whose fibrovascular membranes where used for mRNA analysis.</p

NRTIs reduced nematode body length.

<p>Relative body length was measured after 48hrs NRTI exposure of L1 and 96h exposure of L4 animals (30 worms in ≥2 individual experiments). NRTI concentration = 200μM. Statistics were calculated with a two-way ANOVA with replication, compared to controls. *** = P<0.001.</p

Identification of proteins associated with clinical and pathological features of proliferative diabetic retinopathy in vitreous and fibrovascular membranes

<div><p>Purpose</p><p>To identify the protein profiles in vitreous associated with retinal fibrosis, angiogenesis, and neurite formation in epiretinal fibrovascular membranes (FVMs) in patients with proliferative diabetic retinopathy (PDR).</p><p>Methods</p><p>Vitreous samples of 5 non-diabetic control patients with vitreous debris and 7 patients with PDR membranes were screened for 507 preselected proteins using the semi-quantitative RayBio® L-series 507 antibody array. From this array, 60 proteins were selected for a custom quantitative antibody array (Raybiotech, Human Quantibody® array), analyzing 7 control patients, 8 PDR patients with FVMs, and 5 PDR patients without FVMs. Additionally, mRNA levels of proteins of interest were measured in 10 PDR membranes and 11 idiopathic membranes and in retinal tissues and cells to identify possible sources of protein production.</p><p>Results</p><p>Of the 507 proteins screened, 21 were found to be significantly elevated in PDR patients, including neurogenic and angiogenic factors such as neuregulin 1 (NRG1), nerve growth factor receptor (NGFR), placental growth factor (PlGF) and platelet derived growth factor (PDGF). Angiopoietin-2 (Ang2) concentrations were strongly correlated to the degree of fibrosis and the presence of FVMs in patients with PDR. Protein correlation analysis showed PDGF to be extensively co-regulated with other proteins, including thrombospondin-1 and Ang2. mRNA levels of glial-derived and brain/derived neurotrophic factor (GDNF and BDNF) were elevated in PDR membranes. These results were validated in a second study of 52 vitreous samples of 32 PDR patients and 20 control patients.</p><p>Conclusions</p><p>This exploratory study reveals protein networks that potentially contribute to neurite outgrowth, angiogenesis and fibrosis in the formation of fibrovascular membranes in PDR. We identified a possible role of Ang2 in fibrosis and the formation of FVMs, and of the neurotrophic factors NRG1, PDGF and GDNF in neurite growth that occurs in all FVMs in PDR.</p></div

NAC can mitigate the average lifespan increase caused by FLT.

<p>All animals are N2 exposed from L4 at 20°C. Statistics were conducted compared to the DMSO control. FLT concentration = 200μM, NAC concentration = 100μM.</p