Analysis of solvent tolerance in Pseudomonas putida DOT-T1E based on its genome sequence and a collection of mutants.

Work in our laboratory was supported by Fondo Social Europeo and Fondos FEDER from the European Union through project BIO2010-17227 and Junta de Andalucía Proyecto de Excelencia CVI-301

Efficacy and Safety of Nivolumab Plus Ipilimumab in Patients With Advanced Hepatocellular Carcinoma Previously Treated With Sorafenib The CheckMate 040 Randomized Clinical Trial

IMPORTANCE Most patients with hepatocellular carcinoma (HCC) are diagnosed with advanced disease not eligible for potentially curative therapies; therefore, new treatment options are needed. Combining nivolumab with ipilimumab may improve clinical outcomes compared with nivolumab monotherapy. OBJECTIVE To assess efficacy and safety of nivolumab plus ipilimumab in patients with advanced HCC who were previously treated with sorafenib. DESIGN, SETTING, AND PARTICIPANTS CheckMate 040 is a multicenter, open-label, multicohort, phase 1/2 study. In the nivolumab plus ipilimumab cohort, patients were randomized between January 4 and September 26, 2016. Treatment group information was blinded after randomization. Median follow-up was 30.7 months. Data cutoff for this analysis was January 2019. Patients were recruited at 31 centers in 10 countries/territories in Asia, Europe, and North America. Eligible patients had advanced HCC (with/without hepatitis B or C) previously treated with sorafenib. A total of 148 patients were randomized (50 to arm A and 49 each to arms B and C). INTERVENTIONS Patients were randomized 1:1:1 to either nivolumab 1 mg/kg plus ipilimumab 3 mg/kg, administered every 3 weeks (4 doses), followed by nivolumab 240 mg every 2 weeks (arm A); nivolumab 3 mg/kg plus ipilimumab 1 mg/kg, administered every 3 weeks (4 doses), followed by nivolumab 240 mg every 2 weeks (arm B); or nivolumab 3 mg/kg every 2 weeks plus ipilimumab 1 mg/kg every 6 weeks (arm C). MAIN OUTCOMES AND MEASURES Coprimary end points were safety, tolerability, and objective response rate. Duration of response was also measured (investigator assessed with the Response Evaluation Criteria in Solid Tumors v1.1). RESULTS Of 148 total participants, 120 were male (81%). Median (IQR) age was 60 (52.5-66.5). At data cutoff (January 2019), the median follow-up was 30.7 months (IQR, 29.9-34.7). Investigator-assessed objective response rate was 32% (95% CI, 20%-47%) in arm A, 27% (95% CI, 15%-41%) in arm B, and 29% (95% CI, 17%-43%) in arm C. Median (range) duration of response was not reached (8.3-33.7+) in arm A and was 15.2 months (4.2-29.9+) in arm B and 21.7 months (2.8-32.7+) in arm C. Any-grade treatment-related adverse events were reported in 46 of 49 patients (94%) in arm A, 35 of 49 patients (71%) in arm B, and 38 of 48 patients (79%) in arm C; there was 1 treatment-related death (arm A; grade 5 pneumonitis). CONCLUSIONS AND RELEVANCE In this randomized clinical trial, nivolumab plus ipilimumab had manageable safety, promising objective response rate, and durable responses. The arm A regimen (4 doses nivolumab 1 mg/kg plus ipilimumab 3 mg/kg every 3 weeks then nivolumab 240 mg every 2 weeks) received accelerated approval in the US based on the results of this study. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0165887

Assessing the impact of COVID-19 on liver cancer management (CERO-19)

Background & Aims: The coronavirus disease 2019 (COVID-19) pandemic has posed unprecedented challenges to healthcare systems and it may have heavily impacted patients with liver cancer (LC). Herein, we evaluated whether the schedule of LC screening or procedures has been interrupted or delayed because of the COVID-19 pandemic. Methods: An international survey evaluated the impact of the COVID-19 pandemic on clinical practice and clinical trials from March 2020 to June 2020, as the first phase of a multicentre, international, and observational project. The focus was on patients with hepatocellular carcinoma or intrahepatic cholangiocarcinoma, cared for around the world during the first COVID-19 pandemic wave. Results: Ninety-one centres expressed interest to participate and 76 were included in the analysis, from Europe, South America, North America, Asia, and Africa (73.7%,17.1%, 5.3%, 2.6%, and 1.3% per continent, respectively). Eighty-seven percent of the centres modified their clinical practice: 40.8% the diagnostic procedures, 80.9% the screening programme, 50% cancelled curative and/or palliative treatments for LC, and 41.7% modified the liver transplantation programme. Forty-five out of 69 (65.2%) centres in which clinical trials were running modified their treatments in that setting, but 58.1% were able to recruit new patients. The phone call service was modified in 51.4% of centres which had this service before the COVID-19 pandemic (n = 19/37). Conclusions: The first wave of the COVID-19 pandemic had a tremendous impact on the routine care of patients with liver cancer. Modifications in screening, diagnostic, and treatment algorithms may have significantly impaired the outcome of patients. Ongoing data collection and future analyses will report the benefits and disadvantages of the strategies implemented, aiding future decision-making. Lay summary: The coronavirus disease 2019 (COVID-19) pandemic has posed unprecedented challenges to healthcare systems globally. Herein, we assessed the impact of the first wave pandemic on patients with liver cancer and found that routine care for these patients has been majorly disrupted, which could have a significant impact on outcomes. (C) 2021 The Authors. Published by Elsevier B.V. on behalf of European Association for the Study of the Liver (EASL)

Assessing the impact of COVID-19 on liver cancer management (CERO-19).

BACKGROUND & AIMS: The coronavirus disease 2019 (COVID-19) pandemic has posed unprecedented challenges to healthcare systems and it may have heavily impacted patients with liver cancer (LC). Herein, we evaluated whether the schedule of LC screening or procedures has been interrupted or delayed because of the COVID-19 pandemic. METHODS: An international survey evaluated the impact of the COVID-19 pandemic on clinical practice and clinical trials from March 2020 to June 2020, as the first phase of a multicentre, international, and observational project. The focus was on patients with hepatocellular carcinoma or intrahepatic cholangiocarcinoma, cared for around the world during the first COVID-19 pandemic wave. RESULTS: Ninety-one centres expressed interest to participate and 76 were included in the analysis, from Europe, South America, North America, Asia, and Africa (73.7%, 17.1%, 5.3%, 2.6%, and 1.3% per continent, respectively). Eighty-seven percent of the centres modified their clinical practice: 40.8% the diagnostic procedures, 80.9% the screening programme, 50% cancelled curative and/or palliative treatments for LC, and 41.7% modified the liver transplantation programme. Forty-five out of 69 (65.2%) centres in which clinical trials were running modified their treatments in that setting, but 58.1% were able to recruit new patients. The phone call service was modified in 51.4% of centres which had this service before the COVID-19 pandemic (n = 19/37). CONCLUSIONS: The first wave of the COVID-19 pandemic had a tremendous impact on the routine care of patients with liver cancer. Modifications in screening, diagnostic, and treatment algorithms may have significantly impaired the outcome of patients. Ongoing data collection and future analyses will report the benefits and disadvantages of the strategies implemented, aiding future decision-making. LAY SUMMARY: The coronavirus disease 2019 (COVID-19) pandemic has posed unprecedented challenges to healthcare systems globally. Herein, we assessed the impact of the first wave pandemic on patients with liver cancer and found that routine care for these patients has been majorly disrupted, which could have a significant impact on outcomes

Nurses' perceptions of aids and obstacles to the provision of optimal end of life care in ICU

Contains fulltext : 172380.pdf (publisher's version ) (Open Access

Bacterial chemotaxis towards aromatic hydrocarbons in Pseudomonas

12 pages, 5 figures, 2 tables, 48 references.Bacterial chemotaxis is an adaptive behaviour, which requires sophisticated information-processing capabilities that cause motile bacteria to either move towards or flee from chemicals. Pseudomonas putida DOT-T1E exhibits the capability to move towards different aromatic hydrocarbons present at a wide range of concentrations. The chemotactic response is mediated by the McpT chemoreceptor encoded by the pGRT1 megaplasmid. Two alleles of mcpT are borne on this plasmid and inactivation of either one led to loss of this chemotactic phenotype. Cloning of mcpT into a plasmid complemented not only the mcpT mutants but also its transfer to other Pseudomonas conferred chemotactic response to high concentrations of toluene and other chemicals. Therefore, the phenomenon of chemotaxis towards toxic compounds at high concentrations is gene-dose dependent. In vitro experiments show that McpT is methylated by CheR and McpT net methylation was diminished in the presence of hydrocarbons, what influences chemotactic movement towards these chemicals.Peer Reviewe

Chemotaxis of the human pathogen Pseudomonas aeruginosa to the neurotransmitter Acetylcholine

Acetylcholine is a central biological signal molecule present in all kingdoms of life. In humans, acetylcholine is the primary neurotransmitter of the peripheral nervous system; it mediates signal transmission at neuromuscular junctions. Here, we show that the opportunistic human pathogen Pseudomonas aeruginosa exhibits chemoattraction toward acetylcholine over a concentration range of 1 mM to 100 mM. The maximal magnitude of the response was superior to that of many other P. aeruginosa chemoeffectors. We demonstrate that this chemoattraction is mediated by the PctD (PA4633) chemoreceptor. Using microcalorimetry, we show that the PctD ligand-binding domain (LBD) binds acetylcholine with a equilibrium dissociation constant (KD) of 23 mM. It also binds choline and with lower affinity betaine. Highly sensitive responses to acetylcholine and choline, and less sensitive responses to betaine and L-carnitine, were observed in Escherichia coli expressing a chimeric receptor comprising the PctD-LBD fused to the Tar chemoreceptor signaling domain. We also identified the PacA (ECA_RS10935) chemoreceptor of the phytopathogen Pectobacterium atrosepticum, which binds choline and betaine but fails to recognize acetylcholine. To identify the molecular determinants for acetylcholine recognition, we report high-resolution structures of PctD-LBD (with bound acetylcholine and choline) and PacA-LBD (with bound betaine). We identified an amino acid motif in PctD-LBD that interacts with the acetylcholine tail. This motif is absent in PacA-LBD. Significant acetylcholine chemotaxis was also detected in the plant pathogens Agrobacterium tumefaciens and Dickeya solani. To the best of our knowledge, this is the first report of acetylcholine chemotaxis and extends the range of host signals perceived by bacterial chemoreceptors.This work was supported by grants PID2019-103972GA-I00 (to M.A.M.), PID2020-116261GB-I00 (to J.A.G.) and PID2020-112612GB-I00 (to T.K.) from the Spanish Ministry for Science and Innovation/Agencia Estatal de Investigación 10.13039/501100011033, grant P18-FR-1621 (to T.K.) from the Junta de Andalucía, by the Hessian Ministry of Higher Education, Research, and the Arts (HMWK)–LOEWE research cluster “Diffusible Signals,” subproject A1 (to V.S.), and a Peterson Group “Serving Hometown” Elites scholarship to W.X. We thank Maria Rabyk for help with the hybrid receptor cloning. We are grateful to the European Synchrotron Radiation Facility (ESRF) for the provision of time through proposals MX2281 and MX2353 and to the staff at beamlines ID30B, ID23-1, and ID30A-3 and at the Xaloc beamline of the Alba Spanish synchrotron radiation source (Barcelona) for assistance during data collection

Isotope records (C-O-Sr) of late Pliensbachian-early Toarcian environmental perturbations in the westernmost Tethys (Majorca Island, Spain)

The late Pliensbachian–early Toarcian (Early Jurassic) was a time of major environmental changes that culminated with the Early Toarcian Oceanic Anoxic Event (T-OAE, ca. ~183 Ma). This period is marked by significant disturbances in the carbon cycle and rapid climatic changes. To improve the understanding of the expression of these events in westernmost Tethyan domains, this study provides new belemnite and bulk carbonate C and O stable isotope records of a ~5 Myr upper Pliensbachian to middle Toarcian marine succession of the Balearic Basin (Es Cosconar section, Majorca). Time resolution has been improved by combination of biostratigraphic (ammonoids and brachiopods) and geochronologic (87Sr/86Sr) methods. Seawater paleotemperatures derived from δ18O belemnite records reveal cooler paleotemperatures in the upper part of the Spinatum Zone. The uppermost Spinatum Zone is characterized by the onset of a warming event that crosses the Pliensbachian-Toarcian boundary, culminating with the warmer temperatures (up to ~10 °C of warming) for the Serpentinum Zone of the lower Toarcian. This warming event has been detected contemporaneously in many other European and Tethyan basins and is interpreted to represent generalized raised seawater temperatures linked to the T-OAE. Four significant δ13C events have been recorded in belemnite and bulk carbonate records. The first is a negative carbon isotope excursion (CIE) around the Pliensbachian-Toarcian boundary, which is best represented in the belemnite record. Soon after, the bulk‑carbonate record shows a positive shift in the lower Tenuicostatum Zone concomitant with a return to background values in the belemnite record, suggesting strong water stratification or decoupling probably related with export of neritic carbonate to the basin. The third is a negative CIE represented in bulk carbonate across the Tenuicostatum-Serpentinum zonal transition, which could be correlated with the negative excursion characterizing the onset of the T-OAE in other sections. The position of this excursion corresponds with a gap in the belemnite record. Finally, in the lower Toarcian, both the bulk carbonate and belemnite carbon isotope records show pronounced positive CIEs in the lower-middle part of the Serpentinum Zone. These CIEs testify the impact of the T-OAE in the Balearic basin.Unidad de Tres Cantos, Instituto Geológico y Minero de España, EspañaDepartamento de Paleontología, Universidad Complutense de Madrid, EspañaUnidad de Baleares, Instituto Geológico y Minero de España, Españ

Systematic mapping of chemoreceptor specificities for Pseudomonas aeruginosa

The chemotaxis network, one of the most prominent prokaryotic sensory systems, is present in most motile bacteria and archaea. Although the conserved signaling core of this network is well characterized, ligand specificities of a large majority of diverse chemoreceptors encoded in bacterial genomes remain unknown. Here, we performed a systematic identification and characterization of new chemoeffectors for the opportunistic pathogen Pseudomonas aeruginosa, which has 26 chemoreceptors possessing most of the common types of ligand binding domains. By performing capillary chemotaxis assays for a library of growth-promoting compounds, we first identified a number of novel chemoattractants of varying strength. We subsequently mapped specificities of these ligands by performing Förster resonance energy transfer and microfluidic measurements for 16 hybrid chemoreceptors that combine the periplasmic ligand binding domains of P. aeruginosa receptors and the cytoplasmic signaling domain of the Escherichia coli Tar receptor. Direct binding of putative ligands to chemoreceptors was further confirmed using thermal shift assay and microcalorimetry. Altogether, the combination of methods enabled us to assign several new attractants, including methyl 4-aminobutyrate, 5-aminovalerate, L-ornithine, 2-phenylethylamine, and tyramine, to previously characterized chemoreceptors and to annotate a novel purine-specific receptor PctP. Responses of hybrid receptors to changes in pH further revealed a complex bidirectional pH sensing mechanism in P. aeruginosa, which involves at least four chemoreceptors PctA, PctC, TlpQ, and PctP. Our screening strategy could be applied for the systematic characterization of unknown sensory domains in a wide range of bacterial species.This work was supported by the Max Planck Society and the Hessian Ministry of Higher Education, Research and the Arts (HMWK)–LOEWE research cluster “Diffusible Signals,” subproject A1 (to V.S.), the Spanish Ministry for Science and Innovation/Agencia Estatal de Investigación 10.13039/501100011033 (grants PID2020-112612GB-I00 to T.K. and PID2019-103972GA-I00 to M.A.M.), and the Junta de Andalucía (grant P18-FR-1621 to T.K.). J.P.C.-V. was supported by the grant Unión Europea-Next Generation EU RD 289/2021 UPM-Recualifica Margarita Salas. W.X. was supported by Peterson Group “Serving Hometown” Elites scholarship of Peterson Group Charity Foundation Limited and Chinese Scholarship Council (CSC) scholarship