Effect of neutrophil elastase and its inhibitor EPI-hNE4 on transepithelial sodium transport across normal and cystic fibrosis human nasal epithelial cells

<p>Abstract</p> <p>Background</p> <p>Hyperactivity of the epithelial sodium (Na⁺) channel (ENaC) and increased Na⁺absorption by airway epithelial cells leading to airway surface liquid dehydration and impaired mucociliary clearance are thought to play an important role in the pathogenesis of cystic fibrosis (CF) pulmonary disease. In airway epithelial cells, ENaC is constitutively activated by endogenous trypsin-like serine proteases such as Channel-Activating Proteases (CAPs). It was recently reported that ENaC activity could also be stimulated by apical treatment with human neutrophil elastase (hNE) in a human airway epithelial cell line, suggesting that hNE inhibition could represent a novel therapeutic approach for CF lung disease. However, whether hNE can also activate Na⁺reabsorption in primary human nasal epithelial cells (HNEC) from control or CF patients is currently unknown.</p> <p>Methods</p> <p>We evaluated by short-circuit current (<it>I</it>_sc) measurements the effects of hNE and EPI-hNE4, a specific hNE inhibitor, on ENaC activity in primary cultures of HNEC obtained from control (9) and CF (4) patients.</p> <p>Results</p> <p>Neither hNE nor EPI-hNE4 treatments did modify <it>I</it>_scin control and CF HNEC. Incubation with aprotinin, a Kunitz-type serine protease inhibitor that blocks the activity of endogenous CAPs, decreased <it>I</it>_scby 27.6% and 54% in control and CF HNEC, respectively. In control and CF HNEC pretreated with aprotinin, hNE did significantly stimulate <it>I</it>_sc, an effect which was blocked by EPI-hNE4.</p> <p>Conclusions</p> <p>These results indicate that hNE does activate ENaC and transepithelial Na⁺transport in both normal and CF HNEC, on condition that the activity of endogenous CAPs is first inhibited. The potent inhibitory effect of EPI-hNE4 on hNE-mediated ENaC activation observed in our experiments highlights that the use of EPI-hNE4 could be of interest to reduce ENaC hyperactivity in CF airways.</p

SMAC mimetic Debio 1143 synergizes with taxanes, topoisomerase inhibitors and bromodomain inhibitors to impede growth of lung adenocarcinoma cells

Targeting anti-apoptotic proteins can sensitize tumor cells to conventional chemotherapies or other targeted agents. Antagonizing the Inhibitor of Apoptosis Proteins (IAPs) with mimetics of the pro-apoptotic protein SMAC is one such approach. We used sensitization compound screening to uncover possible agents with the potential to further sensitize lung adenocarcinoma cells to the SMAC mimetic Debio 1143. Several compounds in combination with Debio 1143, including taxanes, topoisomerase inhibitors, and bromodomain inhibitors, super-additively inhibited growth and clonogenicity of lung adenocarcinoma cells. Co-treatment with Debio 1143 and the bromodomain inhibitor JQ1 suppresses the expression of c-IAP1, c-IAP2, and XIAP. Non-canonical NF-κB signaling is also activated following Debio 1143 treatment, and Debio 1143 induces the formation of the ripoptosome in Debio 1143-sensitive cell lines. Sensitivity to Debio 1143 and JQ1 co-treatment was associated with baseline caspase-8 expression. In vivo treatment of lung adenocarcinoma xenografts with Debio 1143 in combination with JQ1 or docetaxel reduced tumor volume more than either single agent alone. As Debio 1143-containing combinations effectively inhibited both in vitro and in vivo growth of lung adenocarcinoma cells, these data provide a rationale for Debio 1143 combinations currently being evaluated in ongoing clinical trials and suggest potential utility of other combinations identified here

DEB025 (Alisporivir) Inhibits Hepatitis C Virus Replication by Preventing a Cyclophilin A Induced Cis-Trans Isomerisation in Domain II of NS5A

DEB025/Debio 025 (Alisporivir) is a cyclophilin (Cyp)-binding molecule with potent anti-hepatitis C virus (HCV) activity both in vitro and in vivo. It is currently being evaluated in phase II clinical trials. DEB025 binds to CypA, a peptidyl-prolyl cis-trans isomerase which is a crucial cofactor for HCV replication. Here we report that it was very difficult to select resistant replicons (genotype 1b) to DEB025, requiring an average of 20 weeks (four independent experiments), compared to the typically <2 weeks with protease or polymerase inhibitors. This indicates a high genetic barrier to resistance for DEB025. Mutation D320E in NS5A was the only mutation consistently selected in the replicon genome. This mutation alone conferred a low-level (3.9-fold) resistance. Replacing the NS5A gene (but not the NS5B gene) from the wild type (WT) genome with the corresponding sequence from the DEB025res replicon resulted in transfer of resistance. Cross-resistance with cyclosporine A (CsA) was observed, whereas NS3 protease and NS5B polymerase inhibitors retained WT-activity against DEB025res replicons. Unlike WT, DEB025res replicon replicated efficiently in CypA knock down cells. However, DEB025 disrupted the interaction between CypA and NS5A regardless of whether the NS5A protein was derived from WT or DEB025res replicon. NMR titration experiments with peptides derived from the WT or the DEB025res domain II of NS5A corroborated this observation in a quantitative manner. Interestingly, comparative NMR studies on two 20-mer NS5A peptides that contain D320 or E320 revealed a shift in population between the major and minor conformers. These data suggest that D320E conferred low-level resistance to DEB025 probably by reducing the need for CypA-dependent isomerisation of NS5A. Prolonged DEB025 treatment and multiple genotypic changes may be necessary to generate significant resistance to DEB025, underlying the high barrier to resistance

Approche diagnostique de l'hyponatrémie [Hyponatremia: diagnostic approach].

Hyponatremia is the most common electrolyte disorder in hospitalized patients and may be associated with non negligible morbidity and mortality. Here we summerize its diagnostic approach based on the physiopathology. Assessment of volume status, measurement of plasma and urinary osmolality remain key steps in the management of this electrolyte disorder

Preoperative Clinical Factors Associated with Short-Stay Laparoscopic Appendectomy.

Outpatient appendectomy for acute appendicitis is a feasible, yet not widely performed procedure, as there are no universally accepted criteria for patient selection. The aim of this study was to assess preoperative clinical factors associated with successful short-stay appendectomy (SSA) and establish a predictive score to help with patient selection. All consecutive laparoscopic appendectomies performed in our institution between January 2013 and June 2015 were retrospectively analyzed. Several preoperative clinical and biological variables were compared between patients with SSA, defined as a postoperative stay &lt;24 h, and those needing inpatient care. Logistic regression analysis was used to identify variables independently associated with SSA, and these variables were then used to create a predictive score. A total of 578 patients were included, 303 (53%) in the SSA group and 275 (48%) in the long-stay appendectomy (LSA) group. In multivariate analysis, male gender (OR 1.61, 95% CI 1.12-2.31, p = 0.010), ASA class I-II (OR 9.52, 95% CI 1.65-180.69, p = 0.037), absence of generalized guarding (OR 3.55, 95% CI 1.30-11.41, p = 0.019), C-reactive protein &lt;100 mg/dl (OR 3.09, 95% CI 1.81-5.42, p &lt; 0.001) and leukocyte count &lt;20 g/l (OR 2.06, 95% CI 1.02-4.30, p = 0.046) were independently associated with SSA. These five parameters were used to construct a predictive score, whereby ≥17 (range 0-21) was defined as the optimal threshold to predict SSA with a high sensitivity (95.6%) and negative predictive value (82.2%). A purely clinical predictive score based on five widely used preoperative parameters can be used to identify eligible patients for short-stay appendectomy

Systematic review of pressurized intraperitoneal aerosol chemotherapy for the treatment of advanced peritoneal carcinomatosis.

Pressurized intraperitoneal aerosol chemotherapy (PIPAC) is a minimally invasive approach under investigation as a novel treatment for patients with peritoneal carcinomatosis of various origins. The aim was to review the available evidence on mechanisms, clinical effects and risks. This was a systematic review of the literature on pressurized intraperitoneal chemotherapy published between January 2000 and October 2016. All types of scientific report were included. Twenty-nine relevant papers were identified; 16 were preclinical studies and 13 were clinical reports. The overall quality of the clinical studies was modest; five studies were prospective and there was no randomized trial. Preclinical data suggested better distribution and higher tissue concentrations of chemotherapy agents in PIPAC compared with conventional intraperitoneal chemotherapy by lavage. Regarding technical feasibility, laparoscopic access and repeatability rates were 83-100 and 38-82 per cent. Surgery-related complications occurred in up to 12 per cent. Postoperative morbidity was low (Common Terminology Criteria for Adverse Events grade 3-5 events reported in 0-37 per cent), and hospital stay was about 3 days. No negative impact on quality of life was reported. Histological response rates for therapy-resistant carcinomatosis of ovarian, colorectal and gastric origin were 62-88, 71-86 and 70-100 per cent respectively. PIPAC is feasible, safe and well tolerated. Preliminary good response rates call for prospective analysis of oncological efficacy

Recovery to Usual Activity After Outpatient Anorectal Surgery.

Most elective anorectal procedures are performed in an outpatient setting, and the supposed recovery time is short. The aim of the present study was to assess return to usual physical activity (UPA), return to work and quality of life (QOL). This prospective single-center cohort study included consecutive patients undergoing outpatient anorectal procedures. Physical and work activities were assessed using the validated International Physical Activity Questionnaire 7 days before surgery and 7, 14 and 30 days thereafter. In addition, patients were inquired daily on their postoperative QOL until postoperative day (POD)10 on a visual analogue scale (0-10). Patients were stratified by their preoperative physical activity score (POPAS; low, moderate and high). Out of 379 patients, 100 (63 men) were included with a median age of 40 years [interquartile range (IQR) 27]. General QOL was rated at a median of 8/10 (IQR 3.5) at POD10. On POD30, only 69% and 71% of patients had returned to UPA and work, respectively. Patients who returned to UPA at POD30 had a better median QOL at POD10 than those who did not (9 vs. 7/10, p = 0.015). Patients with low POPAS and moderate POPAS returned to UPA earlier than patients with high POPAS (83%, 86% and 44% on POD30, respectively, p = 0.005). Return to UPA and work after outpatient anorectal surgery took longer than expected despite a good QOL 10 days after surgery. High physical activity was associated with longer recovery time. These elements should be emphasized during preoperative counseling