Role of N-WASP in Endothelial Monolayer Formation and Integrity

Endothelial cells (ECs) form a monolayer that serves as a barrier between the blood and the underlying tissue. ECs tightly regulate their cell-cell junctions, controlling the passage of soluble materials and immune cells across the monolayer barrier. We studied the role of N-WASP, a key regulator of Arp2/3 complex and actin assembly, in EC monolayers. We report that N-WASP regulates endothelial monolayer integrity by affecting the organization of cell junctions. Depletion of N-WASP resulted in an increase in transendothelial electrical resistance, a measure of monolayer integrity. N-WASP depletion increased the width of cell-cell junctions and altered the organization of F-actin and VE-cadherin at junctions. N-WASP was not present at cell-cell junctions in monolayers under resting conditions, but it was recruited following treatment with sphingosine-1-phosphate. Taken together, our results reveal a novel role for N-WASP in remodeling EC junctions, which is critical for monolayer integrity and function.     

Lack of reversal effect of EDTA treatment on cadmium induced renal dysfunction: a fourteen-year follow-up

The aim of this study was to evaluate the effect of ethylenediaminetetraacetic acid (EDTA) on cadmium (Cd) induced renal dysfunction. Seventeen workers (14 males, 3 females) were diagnosed with occupational Cd poisoning in 1986. These individuals had between 7 to 39 years of Cd exposure. From 1986 to 1999, patients received periodic EDTA therapy as part of their follow-up, all at the same hospital. Levels of urinary cadmium (UCd) and urinary beta2-microglobulin (B2M) were measured before and after each annual EDTA treatment period. Renal dysfunction was defined as urinary B2M > 0.8 mg/g Cr (creatinine). In these workers, patients with UCd level higher than 10 microg/g Cr in 1986 had abnormal B2M excretions (> or = 0.8 mg/g Cr) or trended to have abnormal B2M levels during the treatment period. However, in subjects with UCd concentration lower than 10 microg/g Cr in 1986, their urinary B2M excretions either remained normal (< 0.8 mg/g Cr) or returned to normal during the treatment period. The prevalence of renal dysfunction increased during the follow up period regardless of whether UCd levels increased or not, indicating a progressive renal dysfunction despite removal from Cd exposure. Our results suggest that reversibility of renal dysfunction caused by Cd related to the level of Cd exposure at the time of removal from exposure: renal dysfunction could be reversed if initial UCd < 10 microg/g Cr, but was irreversible when UCd > 10 microg/g Cr. Repeated examinations on these 17 Cd exposed workers from 1986 to 1999 also revealed that periodic administration of EDTA had no beneficial effects on chronic Cd-induced renal dysfunction.     

Pelvic Floor Dysfunction in Morbidly Obese Women: Pilot Study

Objective: The objective of this study was to evaluate the impact of obesity on pelvic floor function in women. Research Methods and Procedures: This was a prospective controlled study of 20 morbidly obese female patients planning to undergo gastric bypass surgery and 20 age‐matched female controls. Subjects completed symptom and impact questionnaires, including the Incontinence Impact Questionnaire (IIQ‐7), Urogenital Distress Inventory (UDI), the Kobashi Prolapse Symptom Inventory and Quality‐of‐Life Questionnaire (PSI‐QOL), and Index of Female Sexual Function. Data were analyzed with Wilcoxon or ratio χ² tests. Results: Mean weight was 295.7 ± 87.9 lbs in the study group and 144.79 ± 33.07 lbs in the control group. Mean BMI was 52.65 ±14.49 kg/m² in the study group and 25.11 ± 5.27 kg/m² in the control group. According to the IIQ‐7, urinary incontinence significantly affected lifestyle in the study group. The total IIQ‐7 score was also significantly affected in the study group (p = 0.03). The UDI indicated more urinary leakage with activity (p = 0.04) and more incidents of small amounts of leakage (p = 0.02) in the study group. According to the PSI‐QOL, women in the study group experienced constipation more often because of difficulty in emptying the rectum (p = 0.04). The PSI‐QOL score was higher in the study group (6.75 ± 6.84) than in the control group (2.65 ± 3.03; p = 0.04). There were no significant differences between groups regarding sexual function. Discussion: Morbid obesity is associated with a significant negative impact on urogenital health. Sexual function did not seem to be affected in women who are morbidly obese.     

Novel 5-HT7R antagonists,arylsulfonamide derivatives of (aryloxy)propyl piperidines: Add-on effect to the antidepressant activity of SSRI and DRI,and pro-cognitive profile

A novel series of arylsulfonamide derivatives of (aryloxy)propyl piperidines was designed to obtain potent 5-HT₇R antagonists. Among the compounds evaluated herein, 3-chloro-N-{1-[3-(1,1-biphenyl-2-yloxy)2-hydroxypropyl]piperidin-4-yl}benzenesulfonamide (25) exhibited antagonistic properties at 5-HT₇R and showed selectivity over selected serotoninergic and dopaminergic receptors, as well as over serotonin, noradrenaline and dopamine transporters. Compound 25 demonstrated significant antidepressant-like activity in the forced swim test (0.625–2.5 mg/kg, i.p.) and in the tail suspension test (1.25 mg/kg, i.p.), augmented the antidepressant effect of inactive doses of escitalopram (selective serotonin reuptake inhibitor) and bupropion (dopamine reuptake inhibitor) in the FST in mice, and similarly to SB-269970, exerted pro-cognitive properties in the novel object recognition task in cognitively unimpaired conditions in rats (0.3 mg/kg, i.p.). Such an extended pharmacological profile, especially the augmentation effect of the identified 5-HT₇R antagonist on SSRI activity, seems promising regarding the complexity of affective disorders and potentially improved outcomes, including mnemonic performance.     

Transgenic rescue of Atg5-null mice from neonatal lethality with neuron-specific expression of ATG5: Systemic analysis of adult Atg5-deficient mice

Atg5-null mice are neonatal lethal. We have revealed in our recent paper that these mice die due to neuronal dysfunction resulting in suckling failure. Our new mouse model, atg5^–/–;Eno2/Nse-Atg5 mice, where Atg5 is deficient in the whole body except for neurons, enables us to analyze the consequences of macroautophagy/autophagy-deficiency in the whole body of adult mice.     

Carbenoxolone inhibits TRPV4 channel‐initiated oxidative urothelial injury and ameliorates cyclophosphamide‐induced bladder dysfunction

Carbenoxolone (CBX) is a clinically prescribed drug for the treatment of digestive ulcer and inflammation. It is also a widely used pharmacological inhibitor of several channels in basic research. Given that the overactivity of several channels, including those inhibitable by CBX, underlies bladder dysfunction, we tested the potential therapeutic application and mechanism of CBX in the treatment of voiding dysfunction. In a mouse model of cystitis induced by cyclophosphamide (CYP), CBX administration prevented the CYP‐elicited increase in bladder weight, oedema, haemorrhage, and urothelial injury. CBX also greatly improved micturition pattern, as manifested by the apparently decreased micturition frequency and increased micturition volume. Western blot results showed that CBX suppressed CYP‐induced increase in protein carbonyls, COX‐2, and iNOS. Further analysis using cultured urothelial cells revealed that acrolein, the major metabolite of CYP, caused protein oxidation, p38 activation, and urothelial injury. These effects of acrolein were reproduced by TRPV4 agonists and significantly prevented by antioxidant NAC, p38 inhibitor SB203580, TRPV4 antagonist RN‐1734, and CBX. Further studies showed that CBX potently suppressed TRPV4 agonist‐initiated calcium influx and subsequent cell injury. CBX attenuated CYP‐induced cystitis in vivo and reduced acrolein‐induced cell injury in vitro, through mechanisms involving inhibition of TRPV4 channels and attenuation of the channel‐mediated oxidative stress. CBX might be a promising agent for the treatment of bladder dysfunction.     

Role of endothelial‐to‐mesenchymal transition induced by TGF‐β1 in transplant kidney interstitial fibrosis

Chronic allograft dysfunction (CAD) induced by kidney interstitial fibrosis is the main cause of allograft failure in kidney transplantation. Endothelial‐to‐mesenchymal transition (EndMT) may play an important role in kidney fibrosis. We, therefore, undertook this study to characterize the functions and potential mechanism of EndMT in transplant kidney interstitial fibrosis. Proteins and mRNAs associated with EndMT were examined in human umbilical vein endothelial cells (HUVECs) treated with transforming growth factor‐beta1 (TGF‐β1) at different doses or at different intervals with western blotting, qRT‐PCR and ELISA assays. Cell motility and migration were evaluated with motility and migration assays. The mechanism of EndMT induced by TGF‐β1 was determined by western blotting analysis of factors involved in various canonical and non‐canonical pathways. In addition, human kidney tissues from control and CAD group were also examined for these proteins by HE, Masson's trichrome, immunohistochemical, indirect immunofluorescence double staining and western blotting assays. TGF‐β1 significantly promoted the development of EndMT in a time‐dependent and dose‐dependent manner and promoted the motility and migration ability of HUVECs. The TGF‐β/Smad and Akt/mTOR/p70S6K signalling pathways were found to be associated with the pathogenesis of EndMT induced by TGF‐β1, which was also proven in vivo by the analysis of specimens from the control and CAD groups. EndMT may promote transplant kidney interstitial fibrosis by targetting the TGF‐β/Smad and Akt/mTOR/p70S6K signalling pathways, and hence, result in the development of CAD in kidney transplant recipients.