Journey on VX-809-Based Hybrid Derivatives towards Drug-like F508del-CFTR Correctors: From Molecular Modeling to Chemical Synthesis and Biological Assays

open12Cystic fibrosis (CF) is a genetic disease affecting the lungs and pancreas and causing progressive damage. CF is caused by mutations abolishing the function of CFTR, a protein whose role is chloride's mobilization in the epithelial cells of various organs. Recently a therapy focused on small molecules has been chosen as a main approach to contrast CF, designing and synthesizing compounds acting as misfolding (correctors) or defective channel gating (potentiators). Multi-drug therapies have been tested with different combinations of the two series of compounds. Previously, we designed and characterized two series of correctors, namely, hybrids, which were conceived including the aminoarylthiazole (AAT) core, merged with the benzodioxole carboxamide moiety featured by VX-809. In this paper, we herein proceeded with molecular modeling studies guiding the design of a new third series of hybrids, featuring structural variations at the thiazole moiety and modifications on position 4. These derivatives were tested in different assays including a YFP functional assay on models F508del-CFTR CFBE41o-cells, alone and in combination with VX-445, and by using electrophysiological techniques on human primary bronchial epithelia to demonstrate their F508del-CFTR corrector ability. This study is aimed (i) at identifying three molecules (9b, 9g, and 9j), useful as novel CFTR correctors with a good efficacy in rescuing the defect of F508del-CFTR; and (ii) at providing useful information to complete the structure-activity study within all the three series of hybrids as possible CFTR correctors, supporting the development of pharmacophore modelling studies, taking into account all the three series of hybrids. Finally, in silico evaluation of the hybrids pharmacokinetic (PK) properties contributed to highlight hybrid developability as drug-like correctors.openParodi, Alice; Righetti, Giada; Pesce, Emanuela; Salis, Annalisa; Tomati, Valeria; Pastorino, Cristina; Tasso, Bruno; Benvenuti, Mirko; Damonte, Gianluca; Pedemonte, Nicoletta; Cichero, Elena; Millo, EnricoParodi, Alice; Righetti, Giada; Pesce, Emanuela; Salis, Annalisa; Tomati, Valeria; Pastorino, Cristina; Tasso, Bruno; Benvenuti, Mirko; Damonte, Gianluca; Pedemonte, Nicoletta; Cichero, Elena; Millo, Enric

EuReCa ONE—27 Nations, ONE Europe, ONE Registry A prospective one month analysis of out-of-hospital cardiac arrest outcomes in 27 countries in Europe

AbstractIntroductionThe aim of the EuReCa ONE study was to determine the incidence, process, and outcome for out of hospital cardiac arrest (OHCA) throughout Europe.MethodsThis was an international, prospective, multi-centre one-month study. Patients who suffered an OHCA during October 2014 who were attended and/or treated by an Emergency Medical Service (EMS) were eligible for inclusion in the study. Data were extracted from national, regional or local registries.ResultsData on 10,682 confirmed OHCAs from 248 regions in 27 countries, covering an estimated population of 174 million. In 7146 (66%) cases, CPR was started by a bystander or by the EMS. The incidence of CPR attempts ranged from 19.0 to 104.0 per 100,000 population per year. 1735 had ROSC on arrival at hospital (25.2%), Overall, 662/6414 (10.3%) in all cases with CPR attempted survived for at least 30 days or to hospital discharge.ConclusionThe results of EuReCa ONE highlight that OHCA is still a major public health problem accounting for a substantial number of deaths in Europe.EuReCa ONE very clearly demonstrates marked differences in the processes for data collection and reported outcomes following OHCA all over Europe. Using these data and analyses, different countries, regions, systems, and concepts can benchmark themselves and may learn from each other to further improve survival following one of our major health care events

EXTRACTION, ANALYSIS AND CHARACTERIZATION OF PRIMARY AND SECONDARY METABOLITES OF THE BASIDIOMYCETE PLEUROTUS OSTREATUS (Jacq.) P. KUMM., 1871, USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY TECHNIQUES COUPLED WITH MASS SPECTROMETRY AND THEIR POSSIBLE USE FOR HUMAN HEALTH

Oxidative stress is one of the main causes of premature cellular aging which can be at the origin of numerous acute or chronic diseases that arise with age. New natural sources of molecules with powerful antioxidant properties are one of the possible solutions in the formulation of antioxidant preparations for protective and anti-aging purposes. As harvested fungi are an evergrowing industry the characterization of their primary and secondary metabolites and their experimentation on cell cultures are fundamental prerequisites for the identification and subsequent experimentation of molecules of cosmetological, pharmaceutical and / or nutraceurtic interest. The aim of this work was the development of a liquid chromatography method at high pressure / performance coupled with mass spectrometry that allowed the simultaneous analysis of as many primary and secondary metabolites extracted from Pleurotus ostreatus as possible. The metabolites identified were tested on cell cultures to highlight their possible cytotoxicity and their possible applications for human health

Comet-FISH analysis of urothelial cells. A screening opportunity for bladder cancer?

Introduction: Bladder cancer (BCa) is the most frequent cancer of the urinary tract, with more than 500,000 reported cases and nearly 200,000 related deaths yearly. Cystoscopy is the standard examination used for the initial diagnosis and follow-up of BCa in the noninvasive stage. However, the American Cancer Society does not include BCa screening in its list of recommended cancer screenings.Areas covered: Recently, several urine-based bladder tumor markers (UBBTMs) that identify genomic, transcriptomic, epigenetic, or protein alterations have been introduced, some of which have been approved by the Food and Drug Administration (FDA) to improve its diagnosis and surveillance. Several biomarkers have been found in the tissues and blood of individuals with BCa or predisposed to develop the disease, further enriching our information.Expert opinion: From a prevention perspective, alkaline Comet-FISH could be a valuable tool with broad potential for clinical application. Furthermore, a comet assay could be more beneficial for diagnosing and monitoring bladder cancer and determining individual susceptibility. Thus, we recommend further studies to understand the potential of this combined assay in the general population as a potential screening test and in patients initiated into the diagnostic process

"Closing the loop\u201d with Oyster Mushroom to recycle agricultural organic waste

Circular economy aims at optimizing production process exploiting and reusing various wastes. Fungi play a key role in recycling lignocellulosic matter and thanks to their ability to produce several metabolite are the most promising organisms to \u201cclosing the loop\u201d in several agricultural production processes. Furthermore, edible fungi are intensely used as nutraceutical and functional foods thanks to their ability to produce bioactive molecules such as antioxidants, \u3b2-glucans, triterpenoids. Our study, framed in the European ALCOTRA project FINNOVER, has two aims: i) exploiting Pleurotus ostreatus to reuse the lavender solid waste from the production of lavender essential oil (extracted by steam distillation); ii) find new products to revamp the agriculture in the western Liguria coast. More precisely, Pulsed Ultrasound-Assisted Extraction (PUAE) was employed to extract from 2 lavender solid wastes (Lavandula angustifolia Mill. and a hybrid called Boscomare) the liquid phenolic compounds whose valorisation as bioactive fraction is another goal of FINNOVER project not discussed here. After this extraction, a Second Solid Waste remains (SSW) was tested for mushrooms cultivations. Two strains of P. ostreatus (one of which is autochthonous from Liguria) were selected to test different substrates in order to find the best spawn recipe for growing P. ostreatus on two different SSWs. More precisely we tested 5 types of substrates made with 2 SSWs mixed with 3 vegetal wastes (poplar sawdust, oak sawdust, and straw) in different percentages. The recipe with the major growth yield was used to prepare a spawn for a laboratory pilot experiment and, later, for a farm pilot experiment. The mycelium was treated for extracting different kinds of metabolites. Once individuated and standardized the extraction, we performed an analysis in HPLC coupled to medium and high resolution mass spectrometry for the quali-quantitative characterization of metabolites, being our work devoted to identify molecules of nutracetical and/or pharmacological interest. The results related in vitro tests showed that lavender up to 30% in the recipe did not affect the miceliar growth rate. Other cultivation pilot tests confirmed these data and highlighted a fruitbody rate of about 20%

Fungi and Circular Economy: Pleurotus ostreatus Grown on a Substrate with Agricultural Waste of Lavender, and Its Promising Biochemical Profile

The increasing production of essential oils has generated a significant amount of vegetal waste that must be discarded, increasing costs for farmers. In this context, fungi, due to their ability to recycle lignocellulosic matter, may be used to turn this waste into new products, thus generating additional income for essential oil producers. The objectives of our work, within the framework of the European ALCOTRA project FINNOVER, were two-fold. The first was to cultivate Pleurotus ostreatus on solid waste of lavender used for essential oil production. The second was to provide, at the same time, new products that can increase the income of small and medium farms in the Ligurian Italian Riviera. This paper presents two pilot tests in which P. ostreatus was grown on substrates with five different concentrations of lavender waste, ranging from 0 to 100% (w/w). Basidiomata grown on all the substrates and their biochemical profiles were characterized using high-performance liquid chromatography coupled to mass spectrometry. The biochemical analysis of mushrooms proved the presence of molecules with antioxidant and potential pharmacological properties, in particular in mushrooms grown on lavender-enriched substrates. The results open the possibility of producing mushrooms classified as a novel food. Furthermore, the results encourage further experiments aimed at investigating how different substrates positively affect the metabolomics of mushrooms

National adoption of video-assisted thoracoscopic surgery (VATS) lobectomy: The Italian VATS register evaluation

Background: The expertise curve of video-assisted thoracoscopic surgery (VATS) lobectomies still stirs debate and controversy both because of the number of procedures to carry out and of the evaluation of the learning threshold. The purpose of our study was the examination of the variables related to the learning curve of the video-assisted approach, to establish what may be an expression of the technical maturity of the surgeon. Methods: The National Register for VATS lobectomy built in 2013 was used to collect data from 65 Thoracic Surgery Units. Out of more than 3,700 patients enrolled, only information from Units with \ue2\u89\ua5100 VATS lobectomies were retrospectively analysed. Unpaired Student's t-tests, Fisher's exact tests, Pearson's \ucf\u872were applied as needed. Cumulative summative analysis and one-way ANOVA were used to identify the expertise curve of VATS lobectomy. Results: Ten institutions contributed a total of 1,679 patients, who were divided into three uniform groups according to the chronological sequence of surgery. The length of utility incision, the number of dissected lymph nodes and the operative time were not statistically significant (P=0.999, P=0.972 and P=0.307, respectively) among groups. Conversion to thoracotomy and postoperative air leaks occurred in 125 (7.44%) and 109 (6.49%) patients, gradually declined in Group 3 with statistical significance (P=0.048 and P=0.00086). Conclusions: The conversion rate and the percentage of air leaks seem to define the expertise of VATS lobectomy, being linked to the ability to manage more complicated surgical cases or intraoperative adverse events

Standardized uptake value and radiological density attenuation as predictive and prognostic factors in patients with solitary pulmonary nodules: Our experience on 1,592 patients

Background: Multislice computed tomography (MSCT) increased detection of solitary pulmonary nodules (SPNs), changing the management based on radiological and clinical factors. When 18-fluorine fluorodeoxyglucose positron emission tomography combined with computed tomography (18F-FDG-PET/CT) was considered for the evaluation of nodules, the maximum standardized uptake value (SUVmax) more than 2.5 is used frequently as a cut off for malignancy. The purpose of this study is to evaluate SUVmax PET/CT and pulmonary attenuation patterns at MSCT in patients with SPN according to morphological and pathological characteristics of the lesion. Methods: A retrospective study on 1,592 SPN patients was carried out following approval by the Italian Registry of VATS Lobectomies. Results: All patients underwent VATS lobectomy. On histologic examination, 98.1% had primary or second metachronous primary lung cancers. In addition, 10.7% presented occult lymph node metastases (pN1 or pN2) on histological examination. Nodule attenuation on CT was associated with the histology of the lesion (p= 0.030); in particular, pure ground glass opacities (pGGOs) and partially solid nodules were related to adenocarcinomatous histotypes. Conversely, a significant relationship between SUVmax and age, nodule size, pathological node status (pN) was found (P=0.007, P=0.000 and P=0.002 respectively). Conclusions: Nodule attenuation can predict the histology of the lesion whereas SUVmax may relate to the propensity to lymph node metastases. Keywords: Solitary pulmonary nodule (SPN); maximum standardized uptake value; ground glass opacities; lymph node metastases; lung adenocarcinom

Weaning from mechanical ventilation in intensive care units across 50 countries (WEAN SAFE): a multicentre, prospective, observational cohort study

Background Current management practices and outcomes in weaning from invasive mechanical ventilation are poorly understood. We aimed to describe the epidemiology, management, timings, risk for failure, and outcomes of weaning in patients requiring at least 2 days of invasive mechanical ventilation. Methods WEAN SAFE was an international, multicentre, prospective, observational cohort study done in 481 intensive care units in 50 countries. Eligible participants were older than 16 years, admitted to a participating intensive care unit, and receiving mechanical ventilation for 2 calendar days or longer. We defined weaning initiation as the first attempt to separate a patient from the ventilator, successful weaning as no reintubation or death within 7 days of extubation, and weaning eligibility criteria based on positive end-expiratory pressure, fractional concentration of oxygen in inspired air, and vasopressors. The primary outcome was the proportion of patients successfully weaned at 90 days. Key secondary outcomes included weaning duration, timing of weaning events, factors associated with weaning delay and weaning failure, and hospital outcomes. This study is registered with ClinicalTrials.gov, NCT03255109. Findings Between Oct 4, 2017, and June 25, 2018, 10 232 patients were screened for eligibility, of whom 5869 were enrolled. 4523 (77·1%) patients underwent at least one separation attempt and 3817 (65·0%) patients were successfully weaned from ventilation at day 90. 237 (4·0%) patients were transferred before any separation attempt, 153 (2·6%) were transferred after at least one separation attempt and not successfully weaned, and 1662 (28·3%) died while invasively ventilated. The median time from fulfilling weaning eligibility criteria to first separation attempt was 1 day (IQR 0–4), and 1013 (22·4%) patients had a delay in initiating first separation of 5 or more days. Of the 4523 (77·1%) patients with separation attempts, 2927 (64·7%) had a short wean (≤1 day), 457 (10·1%) had intermediate weaning (2–6 days), 433 (9·6%) required prolonged weaning (≥7 days), and 706 (15·6%) had weaning failure. Higher sedation scores were independently associated with delayed initiation of weaning. Delayed initiation of weaning and higher sedation scores were independently associated with weaning failure. 1742 (31·8%) of 5479 patients died in the intensive care unit and 2095 (38·3%) of 5465 patients died in hospital. Interpretation In critically ill patients receiving at least 2 days of invasive mechanical ventilation, only 65% were weaned at 90 days. A better understanding of factors that delay the weaning process, such as delays in weaning initiation or excessive sedation levels, might improve weaning success rates

Weaning from mechanical ventilation in intensive care units across 50 countries (WEAN SAFE): a multicentre, prospective, observational cohort study

Background: Current management practices and outcomes in weaning from invasive mechanical ventilation are poorly understood. We aimed to describe the epidemiology, management, timings, risk for failure, and outcomes of weaning in patients requiring at least 2 days of invasive mechanical ventilation. Methods: WEAN SAFE was an international, multicentre, prospective, observational cohort study done in 481 intensive care units in 50 countries. Eligible participants were older than 16 years, admitted to a participating intensive care unit, and receiving mechanical ventilation for 2 calendar days or longer. We defined weaning initiation as the first attempt to separate a patient from the ventilator, successful weaning as no reintubation or death within 7 days of extubation, and weaning eligibility criteria based on positive end-expiratory pressure, fractional concentration of oxygen in inspired air, and vasopressors. The primary outcome was the proportion of patients successfully weaned at 90 days. Key secondary outcomes included weaning duration, timing of weaning events, factors associated with weaning delay and weaning failure, and hospital outcomes. This study is registered with ClinicalTrials.gov, NCT03255109. Findings: Between Oct 4, 2017, and June 25, 2018, 10 232 patients were screened for eligibility, of whom 5869 were enrolled. 4523 (77·1%) patients underwent at least one separation attempt and 3817 (65·0%) patients were successfully weaned from ventilation at day 90. 237 (4·0%) patients were transferred before any separation attempt, 153 (2·6%) were transferred after at least one separation attempt and not successfully weaned, and 1662 (28·3%) died while invasively ventilated. The median time from fulfilling weaning eligibility criteria to first separation attempt was 1 day (IQR 0-4), and 1013 (22·4%) patients had a delay in initiating first separation of 5 or more days. Of the 4523 (77·1%) patients with separation attempts, 2927 (64·7%) had a short wean (≤1 day), 457 (10·1%) had intermediate weaning (2-6 days), 433 (9·6%) required prolonged weaning (≥7 days), and 706 (15·6%) had weaning failure. Higher sedation scores were independently associated with delayed initiation of weaning. Delayed initiation of weaning and higher sedation scores were independently associated with weaning failure. 1742 (31·8%) of 5479 patients died in the intensive care unit and 2095 (38·3%) of 5465 patients died in hospital. Interpretation: In critically ill patients receiving at least 2 days of invasive mechanical ventilation, only 65% were weaned at 90 days. A better understanding of factors that delay the weaning process, such as delays in weaning initiation or excessive sedation levels, might improve weaning success rates. Funding: European Society of Intensive Care Medicine, European Respiratory Society