Bladder inflammatory transcriptome in response to tachykinins: Neurokinin 1 receptor-dependent genes and transcription regulatory elements

Background Tachykinins (TK), such as substance P, and their neurokinin receptors which are ubiquitously expressed in the human urinary tract, represent an endogenous system regulating bladder inflammatory, immune responses, and visceral hypersensitivity. Increasing evidence correlates alterations in the TK system with urinary tract diseases such as neurogenic bladders, outflow obstruction, idiopathic detrusor instability, and interstitial cystitis. However, despite promising effects in animal models, there seems to be no published clinical study showing that NK-receptor antagonists are an effective treatment of pain in general or urinary tract disorders, such as detrusor overactivity. In order to search for therapeutic targets that could block the tachykinin system, we set forth to determine the regulatory network downstream of NK1 receptor activation. First, NK1R-dependent transcripts were determined and used to query known databases for their respective transcription regulatory elements (TREs). Methods: An expression analysis was performed using urinary bladders isolated from sensitized wild type (WT) and NK1R-/- mice that were stimulated with saline, LPS, or antigen to provoke inflammation. Based on cDNA array results, NK1R-dependent genes were selected. PAINT software was used to query TRANSFAC database and to retrieve upstream TREs that were confirmed by electrophoretic mobility shift assays. Results: The regulatory network of TREs driving NK1R-dependent genes presented cRel in a central position driving 22% of all genes, followed by AP-1, NF-kappaB, v-Myb, CRE-BP1/c-Jun, USF, Pax-6, Efr-1, Egr-3, and AREB6. A comparison between NK1R-dependent and NK1R-independent genes revealed Nkx-2.5 as a unique discriminator. In the presence of NK1R, Nkx2-5 _01 was significantly correlated with 36 transcripts which included several candidates for mediating bladder development (FGF) and inflammation (PAR-3, IL-1R, IL-6, α-NGF, TSP2). In the absence of NK1R, the matrix Nkx2-5_02 had a predominant participation driving 8 transcripts, which includes those involved in cancer (EYA1, Trail, HSF1, and ELK-1), smooth-to-skeletal muscle trans-differentiation, and Z01, a tight-junction protein, expression. Electrophoretic mobility shift assays confirmed that, in the mouse urinary bladder, activation of NK1R by substance P (SP) induces both NKx-2.5 and NF-kappaB translocations. Conclusion: This is the first report describing a role for Nkx2.5 in the urinary tract. As Nkx2.5 is the unique discriminator of NK1R-modulated inflammation, it can be imagined that in the near future, new based therapies selective for controlling Nkx2.5 activity in the urinary tract may be used in the treatment in a number of bladder disorders

Comparative analysis of the ATRX promoter and 5' regulatory region reveals conserved regulatory elements which are linked to roles in neurodevelopment, alpha-globin regulation and testicular function

BACKGROUND ATRX is a tightly-regulated multifunctional protein with crucial roles in mammalian development. Mutations in the ATRX gene cause ATR-X syndrome, an X-linked recessive developmental disorder resulting in severe mental retardation and mild alpha-thalassemia with facial, skeletal and genital abnormalities. Although ubiquitously expressed the clinical features of the syndrome indicate that ATRX is not likely to be a global regulator of gene expression but involved in regulating specific target genes. The regulation of ATRX expression is not well understood and this is reflected by the current lack of identified upstream regulators. The availability of genomic data from a range of species and the very highly conserved 5' regulatory regions of the ATRX gene has allowed us to investigate putative transcription factor binding sites (TFBSs) in evolutionarily conserved regions of the mammalian ATRX promoter. RESULTS We identified 12 highly conserved TFBSs of key gene regulators involved in biologically relevant processes such as neural and testis development and alpha-globin regulation. CONCLUSIONS Our results reveal potentially important regulatory elements in the ATRX gene which may lead to the identification of upstream regulators of ATRX and aid in the understanding of the molecular mechanisms that underlie ATR-X syndrome.This work was supported by Department of Zoology research grants

Investigating the Prevalence of Renal Artery Stenosis Following Coronary Artery Angiography and Related Risks in Hypertensive Patients Candidate for Coronary Artery Angiography in Yazd Afshar Hospital

Introduction: Renal artery stenosis is the most prevalent disease of renal artery and has an important role in making hypertension and renal atrophy. Since in previous researches, despite high frequency of risk of cardiovascular disease in Iran, there did not exist any general research investigating risk factors of coronary artery disease and incidence of contemporary renal artery stenisis, this study intended to investigate these factors in hypertensive patients in Iran. Methods: This cross-sectional study was performed on 264 patients who were hypertension candidate for coronary artery angiography in Afshar hospital in Yazd. At the time of coronary angiography, patients had renal angiography at the same time. Among those patients, those who had coronary artery disease were selected and thus prevalence and severity of renal artery stenosis and its relationship with cardiovascular risks were investigated. The gathered data were analyzed by Spss-18 software. Results: Prevalence of renal artery stenosis (equal and over 50%) was 38.25 in all patients; in other words, in men and women it was 43.56% and 56.44% (P=0.04) respectively. Prevalence of co-morbidity of coronary artery disease and renal artery stenosis with 1 vessel disease was 34%, 2 vessel disease was 57.14% and 3 vessel disease was 54.17% with significant differences (P>0.01) Conclusion: According to high prevalence of renal artery stenosis in hypertensive patients and high prevalency with simultaneous hypertension and coronary artery disease, renal angiography after coronary angiography may assist for unknown renal artery stenosis and better treatment

Effect of glycated insulin on the blood-brain barrier permeability: An in vitro study

© 2018 Altered blood-brain barrier (BBB) permeability may contribute to pathogenesis of diabetes-related central nervous system disorders. Considering the presence of glycated insulin in plasma of type 2 diabetic patients, we hypothesized that glycated insulin could induce changes in paracellular permeability in BBB. Therefore, the authors decided to study the effect of glycated insulin on paracellular permeability in a BBB model and the change induced in insulin conformation upon glycation. In this study, the structural modification was examined by fluorescence and circular dichroism spectroscopies and dynamic light scattering. Cell proliferation and production of ROS in astrocytes and HUVEC cells were analyzed by MTT and spectrofluorometric assays, respectively. Apoptosis induction was determined and confirmed by flow cytometry and western blot analyses, respectively. The permeability was measured Lucifer yellow and FITC-Dextran. According to our results, glycated insulin presented altered conformation and more exposed hydrophobic patches than insulin. Formation of oligomeric species and advanced glycated end products (AGEs) were determined. Lower cell viability, higher apoptosis, and more ROS were detected upon treatment of cells with glycated insulin. Finally, glycated insulin led to increased Lucifer yellow and FITC-dextran transportation across the BBB model which could result from ROS producing and apoptosis-inducing activities of AGE-insulin

"PRIMARY VITRECTOMY VERSUS SCLERAL BUCKLING IN PATIENTS WITH RETINAL DETACHMENT AFTER CATARACT SURGERY"

The purpose of this study was to compare the anatomic and visual outcome of primary vitrectomy with scleral buckling in patients with retinal detachment following cataract surgery. Fifty-six consecutive patients with retinal detachment after cataract surgery were randomly assigned to two treatment groups: standard scleral buckling and standard three-port deep vitrectomy. Successful treatment was defined as improvement in vision (minimum of 2 lines in Snellen chart), anatomic reattachment and prevention of post-operative proliferative vitreo-retinopathy (PVR). The prognostic role of pre-operative and intra-operative conditions of the affected eye was also evaluated. Twenty-six of fifty-six eligible patients underwent scleral buckling and thirty had deep vitrectomy. Anatomic reattachment was achieved in 18 (69.2%) cases in scleral buckling group and 19 (63%) cases in vitrectomy group. Improvement in visual acuity was achieved in 76.9% and 83.3% and PVR occurred post-operatively in 23.1% and 16.7%, respectively. The differences were not statistically significant, and pre- and intra-operative ocular conditions did not prove to be prognostic factors, either. Scleral buckling and primary deep vitrectomy seem to have comparable outcomes in terms of anatomic reattachment and visual improvement in patients with pseudophakic and aphakic retinal detachment. Failure to achieve anatomic reattachment and visual improvement or PVR occurred in about one third and one fifth of the cases respectively, irrespective of the technique used. This warrants further research to improve treatment result