Gene expression pattern in swine neutrophils after lipopolysaccharide exposure: a time course comparison

Background: Experimental exposure of swine neutrophils to bacterial lipopolysaccharide (LPS) represents a model to study the innate immune response during bacterial infection. Neutrophils can effectively limit the infection by secreting lipid mediators, antimicrobial molecules and a combination of reactive oxygen species (ROS) without new synthesis of proteins. However, it is known that neutrophils can modify the gene expression after LPS exposure. We performed microarray gene expression analysis in order to elucidate the less known transcriptional response of neutrophils during infection. Methods: Blood samples were collected from four healthy Iberian pigs and neutrophils were isolated and incubated during 6, 9 and 18 hrs in presence or absence of lipopolysaccharide (LPS) from Salmonella enterica serovar Typhimurium. RNA was isolated and hybridized to Affymetrix Porcine GeneChip®. Microarray data were normalized using Robust Microarray Analysis (RMA) and then, differential expression was obtained by an analysis of variance (ANOVA). Results: ANOVA data analysis showed that the number of differentially expressed genes (DEG) after LPS treatment vary with time. The highest transcriptional response occurred at 9 hr post LPS stimulation with 1494 DEG whereas at 6 and 18 hr showed 125 and 108 DEG, respectively. Three different gene expression tendencies were observed: genes in cluster 1 showed a tendency toward up-regulation; cluster 2 genes showing a tendency for down-regulation at 9 hr; and cluster 3 genes were up-regulated at 9 hr post LPS stimulation. Ingenuity Pathway Analysis revealed a delay of neutrophil apoptosis at 9 hr. Many genes controlling biological functions were altered with time including those controlling metabolism and cell organization, ubiquitination, adhesion, movement or inflammatory response. Conclusions: LPS stimulation alters the transcriptional pattern in neutrophils and the present results show that the robust transcriptional potential of neutrophils under infection conditions, indicating that active regulation of gene expression plays a major role in the neutrophil-mediated- innate immune respons

Alkane oxidation by a carboxylate-bridged dimanganese(III) complex

A new manganese(III) oxamato dimer possesing an unprecedented Mn2(μ-O2CR)(μ-OH2…O2CR) core has been synthesised, structurally and magnetically characterised, and used as a catalyst for the oxidation of alkanes to alcohols and ketones by Bu^tO2H and O2 in CH2Cl2 at rt.Blay Llinares, Gonzalo, [email protected] ; Fernandez Picot, Isabel, [email protected] ; Pedro Llinares, Jose Ramon, [email protected] ; Ruiz Garcia, Rafael, [email protected] ; Pardo Marin, Emilio Jose, [email protected] ; Lloret Pastor, Francisco, [email protected]

Efficacy of novel indoor residual spraying methods targeting pyrethroid-resistant aedes aegypti within experimental houses

Challenges in maintaining high effectiveness of classic vector control in urban areas has renewed the interest in indoor residual spraying (IRS) as a promising approach for Aedes-borne disease prevention. While IRS has many benefits, application time and intrusive indoor applications make its scalability in urban areas difficult. Modifying IRS to account for Ae. aegypti resting behavior, named targeted IRS (TIRS, spraying walls below 1.5 m and under furniture) can reduce application time; however, an untested assumption is that modifications to IRS will not negatively impact entomological efficacy. We conducted a comparative experimental study evaluating the residual efficacy of classically-applied IRS (as developed for malaria control) compared to two TIRS application methods using a carbamate insecticide against a pyrethroid-resistant, field-derived Ae. aegypti strain. We performed our study within a novel experimental house setting (n = 9 houses) located in Merida (Mexico), with similar layouts and standardized contents. Classic IRS application (insecti-cide applied to full walls and under furniture) was compared to: a) TIRS: insecticide applied to walls below 1.5 m and under furniture, and b) Resting Site TIRS (RS-TIRS): insecticide applied only under furniture. Mosquito mortality was measured eight times post-application (out to six months post-application) by releasing 100 Ae. aegypti females/house and collecting live and dead individuals after 24 hrs exposure. Compared to Classic IRS, TIRS and RS-TIRS took less time to apply (31% and 82% reduction, respectively) and used less insecticide (38% and 85% reduction, respectively). Mortality of pyrethroid-resistant Ae. aegypti did not significantly differ among the three IRS application methods up to two months post application, and did not significantly differ between Classic IRS and TIRS up to four months post application. These data illustrate that optimizing IRS to more efficiently target Ae. aegypti

Deltamethrin resistance in Aedes aegypti results in treatment failure in Merida, Mexico

The operational impact of deltamethrin resistance on the efficacy of indoor insecticide applications to control Aedes aegypti was evaluated in Merida, Mexico. A randomized controlled trial quantified the efficacy of indoor residual spraying (IRS) against adult Ae. aegypti in houses treated with either deltamethrin (to which local Ae. aegypti expressed a high degree of resistance) or bendiocarb (to which local Ae. aegypti were fully susceptible) as compared to untreated control houses. All adult Ae. aegypti infestation indices during 3 months post-spraying were significantly lower in houses treated with bendiocarb compared to untreated houses (odds ratio < 0.75; incidence rate ratio < 0.65) whereas no statistically significant difference was detected between the untreated and the deltamethrin-treated houses. On average, bendiocarb spraying reduced Ae. aegypti abundance by 60% during a 3-month period. Results demonstrate that vector control efficacy can be significantly compromised when the insecticide resistance status of Ae. aegypti populations is not taken into consideration

Spanish Multicenter Study of the Epidemiology and Mechanisms of Amoxicillin-Clavulanate Resistance in Escherichia coli

We conducted a prospective multicenter study in Spain to characterize the mechanisms of resistance to amoxicillin-clavu-lanate (AMC) in Escherichia coli. Up to 44 AMC-resistant E. coli isolates (MIC>32/16 g/ml) were collected at each of theseven participant hospitals. Resistance mechanisms were characterized by PCR and sequencing. Molecular epidemiology was studied by pulsed-field gel electrophoresis (PFGE) and by multilocus sequence typing. Overall AMC resistance was 9.3%. The resistance mechanisms detected in the 257 AMC-resistant isolates were OXA-1 production (26.1%), hyperpro-duction of penicillinase (22.6%), production of plasmidic AmpC (19.5%), hyperproduction of chromosomic AmpC(18.3%), and production of inhibitor-resistant TEM (IRT) (17.5%). The IRTs identified were TEM-40 (33.3%), TEM-30(28.9%), TEM-33 (11.1%), TEM-32 (4.4%), TEM-34 (4.4%), TEM-35 (2.2%), TEM-54 (2.2%), TEM-76 (2.2%), TEM-79(2.2%), and the new TEM-185 (8.8%). By PFGE, a high degree of genetic diversity was observed although two well-defined clusters were detected in the OXA-1-producing isolates: the C1 cluster consisting of 19 phylogroup A/sequence type 88[ST88] isolates and the C2 cluster consisting of 19 phylogroup B2/ST131 isolates (16 of them producing CTX-M-15). Each of the clusters was detected in six different hospitals. In total, 21.8% of the isolates were serotype O25b/phylogroup B2 (O25b/B2). AMC resistance in E. coli is widespread in Spain at the hospital and community levels. A high prevalence of OXA-1 was found. Although resistant isolates were genetically diverse, clonality was linked to OXA-1-producing isolates of the STs 88 and 131. Dissemination of IRTs was frequent, and the epidemic O25b/B2/ST131 clone carried many different mechanisms of AMC resistance

Synthesis of Nβ-Substituted α,β-Diamino Acids via Stereoselective N-Michael Additions to a Chiral Bicyclic Dehydroalanine

The highly diastereoselective 1,4-conjugate additions of several nitrogen nucleophiles to chiral bicyclic dehydroalanines have been assessed effectively at room temperature in good to excellent yields without needing any catalyst or additional base. This methodology is general, simple, oxygen and moisture tolerant, high-yielding, totally chemo- and stereoselective. This procedure offers an efficient and practical approach for the synthesis of Nβ-substituted α,β-diamino acids, such as 1-isohistidine, τ-histidinoalanine, β-benzylaminoalanine, β-(piperidin-1-yl)alanine, β-(azepan-1-yl)alanine, and fluorescent and ciprofloxacin-containing amino acid derivatives.We thank the Ministerio de Ciencia, Innovacion y Universidades (RTI2018-099592-B-C21 and RTI2018-099592-B-C22 projects) and the Ministerio de Economia y Competitividad (RYC-2013-14706 to G.J.-O.). N.M. and P.O. thank Universidad de La Rioja for a grant. We also thank Universidad de La Rioja (Beronia cluster) for computer support

Initial experience with rapid deployment Edwards Intuity aortic valve

© 2015 Sociedad Española de Cirugía Torácica-Cardiovascular. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ This document is the Published version of a Published Work that appeared in final form in Cirugía Cardiovascular. To access the final edited and published work see https://doi.org/10.1016/j.circv.2015.07.008Objetivo Disponemos de una nueva clase de prótesis aórticas de despliegue rápido para tratamiento quirúrgico de la estenosis aórtica. Permitirían disminuir los tiempos quirúrgicos y facilitarían la cirugía mínimamente invasiva. Presentamos nuestra experiencia inicial con la válvula Edwards Intuity en el contexto del estudio multicéntrico Foundation. Pacientes y métodos Entre septiembre de 2012 y febrero de 2014 se implantaron 25 prótesis aórticas Edwards Intuity en 26 pacientes (77 ± 4,3 años; 52% varones). Veinticuatro (96%) fueron reemplazo valvular aislado, y 9 pacientes (36%) tuvieron abordaje mínimamente invasivo. Los pacientes fueron estudiados al alta, a los 3 meses y al año de la intervención. Resultados Hubo éxito en el implante en el 96,2% de los pacientes (25/26). No hubo mortalidad hospitalaria. Seguimiento medio de 11 ± 5,4 meses, supervivencia acumulada del 96%. Un paciente falleció a los 11 meses por un ictus isquémico. El tiempo medio de pinzamiento aórtico fue de 38,2 ± 10,1 min y el tiempo medio de circulación extracorpórea, de 62,4 ± 11,1 min. A los 3 meses, el área valvular efectiva media fue de 1,70 ± 0,42 cm2. El 91% de los pacientes (23/25) estaban en grado funcional i en el seguimiento. Conclusiones El implante de la válvula Edwards Intuity para el tratamiento de la estenosis aórtica ha sido factible, seguro y eficaz. Los tiempos de isquemia miocárdica y de circulación extracorpórea parecen reducirse en comparación con la cirugía valvular aórtica habitual. El comportamiento hemodinámico inicial de la prótesis Edwards Intuity es excelente.--------------Objective A new class of rapid deployment prostheses is now available for surgical treatment of aortic stenosis. These prostheses offer the possibility of reducing the duration of cardiopulmonary bypass and aortic clamping times, and facilitate the use of minimally invasive surgery. We present our initial experience with the Edwards Intuity valve in the context of a multicentre Foundation study. Patients and methods Between September 2012 and February 2014, 25 Edwards Intuity valves were implanted in 26 patients (77 ± 4.3 years; 52% male). Twenty-four (96%) were isolated aortic valve replacement; a minimally invasive approach was used on 9 patients (36%). Patients were studied at discharge, three months, and one year after surgery. Results Implantation success rate was 96.2% (25/26). There was no hospital mortality. Mean follow-up of 11 ± 5.54 months, cumulative survival of 96%. One patient died eleven months after surgery due to ischaemic stroke. The mean aortic clamping time was 38.2 ± 10.1 minutes, and the mean cardiopulmonary bypass (CPB) time was 62.4 ± 11.1 minutes. In the echocardiography at 3 months, the mean effective valve area was 1.70 ± 0.42 cm2, and 91% of patients (23/25) were in functional class i NYHA at follow-up. Conclusions The implantation of the Edwards Intuity valve for surgical treatment of aortic stenosis has shown to be feasible, safe and effective. Times of myocardial ischaemia and extracorporeal circulation appear to be reduced compared with normal aortic valve surgery. The initial haemodynamic performance of the Edwards Intuity prosthesis is excellent

Perfusion Decellularization of Extrahepatic Bile Duct Allows Tissue-Engineered Scaffold Generation by Preserving Matrix Architecture and Cytocompatibility

Reconstruction of bile ducts damaged remains a vexing medical problem. Surgeons have few options when it comes to a long segment reconstruction of the bile duct. Biological scaffolds of decellularized biliary origin may offer an approach to support the replace of bile ducts. Our objective was to obtain an extracellular matrix scaffold derived from porcine extrahepatic bile ducts (dECM-BD) and to analyze its biological and biochemical properties. The efficiency of the tailored perfusion decellularization process was assessed through histology stainings. Results from 4’-6-diamidino-2-phenylindole (DAPI), Hematoxylin and Eosin (H&E) stainings, and deoxyribonucleic acid (DNA) quantification showed proper extracellular matrix (ECM) decellularization with an effectiveness of 98%. Immunohistochemistry results indicate an effective decrease in immunogenic marker as human leukocyte antigens (HLA-A) and Cytokeratin 7 (CK7) proteins. The ECM of the bile duct was preserved according to Masson and Herovici stainings. Data derived from scanning electron microscopy (SEM) and thermogravimetric analysis (TGA) showed the preservation of the dECM-BD hierarchical structures. Cytotoxicity of dECM-BD was null, with cells able to infiltrate the scaffold. In this work, we standardized a decellularization method that allows one to obtain a natural bile duct scaffold with hierarchical ultrastructure preservation and adequate cytocompatibility

Circulating Tumor Cells Enumeration from the Portal Vein for Risk Stratification in Early Pancreatic Cancer Patients

[Simple Summary] Effective biomarkers are needed to enable personalized medicine for pancreatic cancer patients. This study analyzes the prognostic value, in early pancreatic cancer, of circulating tumor cells and clusters from the central venous catheter and portal blood. Circulating tumor cells were isolated using an immunomagnetic selection and were detected by microscopy using immunocytochemistry staining. In conclusion, the circulating tumor cell number in portal blood identifies a death risk in patients with early pancreatic cancer.[Abstract] Background. Effective biomarkers are needed to enable personalized medicine for pancreatic cancer patients. This study analyzes the prognostic value, in early pancreatic cancer, of single circulating tumor cell (CTC) and CTC clusters from the central venous catheter (CVC) and portal blood (PV). Methods. In total, 7 mL of PV and CVC blood from 35 patients with early pancreatic cancer were analyzed. CTC were isolated using a positive immunomagnetic selection. The detection and identification of CTC were performed by immunocytochemistry (ICC) and were analyzed by Epi-fluorescence and confocal microscopy. Results. CTC and the clusters were detected both in PV and CVC. In both samples, the CTC number per cluster was higher in patients with grade three or poorly differentiated tumors (G3) than in patients with well (G1) or moderately (G2) differentiated. Patients with fewer than 185 CTC in PV exhibited a longer OS than patients with more than 185 CTC (24.5 vs. 10.0 months; p = 0.018). Similarly, patients with fewer than 15 clusters in PV showed a longer OS than patients with more than 15 clusters (19 vs. 10 months; p = 0.004). These significant correlations were not observed in CVC analyses. Conclusions. CTC presence in PV could be an important prognostic factor to predict poor prognosis in early pancreatic cancer. In addition, the number of clustered-CTC correlate to a tumor negative differentiation degree and, therefore, could be used as a diagnostic biomarker for pancreatic cancer.This research was funded by Carlos III Health Institute (Health Research Fund) grant number PI16/01465 and PI19/01821 (Co-financed by the European Regional Development Fund “A way to make Europe”)