Phosphorylation of B14.5a subunit from bovine heart complex I identified by titanium dioxide selective enrichment and shotgun proteomics

Shotgun proteomics was used to study the steady phosphorylation state of NADH:ubiquinone oxidoreductase (complex I) subunits from bovine heart mitochondria. A total tryptic digestion of enzymatically active complex I was performed, and the resulting peptide mixture was subjected to phosphopeptide enrichment by the use of titanium dioxide (TiO2). The phosphopeptide-enriched fraction was separated and analyzed with nanoscale reverse-phase HPLC-ESI-MS/MS in single information-dependent acquisition. Hence two phosphorylated complex I subunits were detected: 42 kDa and B14.5a. Phosphorylation of 42-kDa subunit at Ser-59 has already been determined with fluorescent phosphoprotein-specific gel staining and mass spectrometry (Schilling, B., Aggeler, R., Schulenberg, B., Murray, J., Row, R. H., Capaldi, R. A., and Gibson, B. W. (2005) Mass spectrometric identification of novel phosphorylation site in subunit NDUFA10 of bovine mitochondrial complex I. FEBS Lett. 579, 2485-2490). In our work, this finding was confirmed using a non-gel-based approach. In addition, we report novel phosphorylation on B14.5a nuclear encoded subunit. We demonstrated evidence of the phosphorylation site at Ser-95 residue by collision-induced dissociation experiments on three different molecular ions of two tryptic phosphopeptides of B14.5a

Vapor etching to avoid micro-masking by gas-bubbles in wet release of MEMS

We report on the formation of gas bubbles during the release of MEMS devices using buffered oxide etch. Several approaches to mitigate the problem are proposed and tested together with a qualitative study of the phenomenon. The chemical reaction behind such phenomenon and the influence of defects and topography is discussed. Finally, a comparison with the HF-vapor release technique is shown

Clinical and environmental distribution of legionella pneumophila in a university hospital in italy: efficacy of ultraviolet disinfection

The molecular epidemiology of Legionella pneumophila in the 'V. Monaldi' University Hospital was studied. Seven cases of nosocomial Legionnaires' disease were diagnosed between 1999 and 2003. Two clinical legionella strains obtained from two patients in the adult cardiac surgery unit (CSU) and 30 environmental legionella strains from the paediatric and adult CSUs, neonatal intensive care unit (NICU) and the cardiorespiratory intensive care unit (CR-ICU) were serotyped and genotyped. L. pneumophila serogroup 1/Philadelphia with an identical pulsed-field gel electrophoresis (PFGE) profile A was isolated from two patients in the adult CSU, and from three and one water samples taken in the adult CSU and the paediatric CSU, respectively, from 2001 to 2002. Furthermore, L. pneumophila serogroup 3 with an identical PFGE profile B was identified in 20 environmental strains from all wards, L. pneumophila serogroup 3 with PFGE profile C was identified in a single environmental strain from the CR-ICU, and non-pneumophila Legionella with identical PFGE profile D was identified in five environmental strains from the adult CSU, paediatric CSU and NICU. Ultraviolet irradiation was effective in disinfection of the hospital water supplies in the adult and paediatric CSUs contaminated by L. pneumophila clone associated with nosocomial Legionnaires' disease. In conclusion, these data demonstrate that two cases of nosocomial legionellosis were caused by the persistence of a single clone of L. pneumophila serogroup 1/Philadelphia in the hospital environment, and that disinfection by ultraviolet irradiation may represent an effective measure to prevent nosocomial Legionnaires' disease. © 2005 The Hospital Infection Society. Published by Elsevier Ltd. All rights reserved

An evidence map and synthesis review with meta-analysis on the risk of incisional hernia in colorectal surgery with standard closure.

Purpose To assess the incidence of incisional hernia (IH) across various type of incisions in colorectal surgery (CS) creating a map of evidence to define research trends, gaps and areas of future interest. Methods Systematic review of PubMed and Scopus from 2010 onwards. Studies included both open (OS) and laparoscopic (LS). The primary outcome was incidence of IH 12 months after index procedure, secondary outcomes were the study features and their influence on reported proportion of IH. Random effects models were used to calculate pooled proportions. Meta-regression models were performed to explore heterogeneity. Results Ninetyone studies were included reporting 6473 IH. The pooled proportions of IH for OS were 0.35 (95% CI 0.27–0.44) I2 0% in midline laparotomies and 0.02 (95% CI 0.00–0.07), I2 52% for off-midline. In case of LS the pooled proportion of IH for midline extraction sites were 0.10 (95% CI 0.07–0.16), I2 58% and 0.04 (95% CI 0.03–0.06), I2 86% in case of off-midline. In Port-site IH was 0.02 (95% CI 0.01–0.04), I2 82%, and for single incision surgery (SILS) of 0.06—95% CI 0.02–0.15, I2 81%. In case of stoma reversal sites was 0.20 (95% CI 0.16–0.24). Conclusion Midline laparotomies and stoma reversal sites are at high risk for IH and should be considered in research of preventive strategies of closure. After laparoscopic approach IH happens mainly by extraction sites incisions specially midline and also represent an important area of analysis.pre-print3102 K

Detection and characterization of classical and "uncommon" exon 19 Epidermal Growth Factor Receptor mutations in lung cancer by pyrosequencing

BACKGROUND: The management of advanced stage non-small cell lung cancer is increasingly based on diagnostic and predictive analyses performed mostly on limited amounts of tumor tissue. The evaluation of Epidermal Growth Factor Receptor (EGFR) mutations have emerged as the strongest predictor of response to EGFR-tyrosine kinase inhibitors mainly in patients with adenocarcinoma. Several EGFR mutation detection techniques are available, having both sensitivity and specificity issues, being the Sanger sequencing technique the reference standard, with the limitation of a relatively high amount of mutated cells needed for the analysis. METHODS: A novel nucleotide dispensation order for pyrosequencing was established allowing the identification and characterization of EGFR mutation not definable with commercially and clinically approved kits, and validated in a consecutive series of 321 lung cancer patients (246 biopsies or cytology samples and 75 surgical specimens). RESULTS: 61/321 (19%) mutated cases were detected, 17 (27.9%) in exon 21 and 44 (72.1%) in exon 19, these latter corresponding to 32/44 (72.7%) classical and 12/44 (27.3%) uncommon mutations. Furthermore, a novel, never reported, point mutation, was found, which determined a premature stop codon in the aminoacidic sequence that resulted in a truncated protein in the tyrosine kinase domain, thus impairing the inhibitory effect of specific therapy. CONCLUSIONS: The novel dispensation order allows to detect and characterize both classical and uncommon EGFR mutations. Although several phase III studies in genotypically defined groups of patients are already available, further prospective studies assessing the role of uncommon EGFR mutations are warranted

Impact of neoadjuvant therapy followed by laparoscopic radical gastrectomy with D2 lymph node dissection in Western population: A multi-institutional propensity score-matched study

Background and Objectives: In the setting of a minimally invasive approach, we aimed to compare short and long-term postoperative outcomes of patients treated with neoadjuvant therapy (NAT) + surgery or upfront surgery in Western population. Methods: All consecutive patients from six Italian and one Serbian center with locally advanced gastric cancer who had undergone laparoscopic gastrectomy with D2 lymph node dissection were selected between 2005 and 2019. After propensity score-matching, postoperative morbidity and oncologic outcomes were investigated. Results: After matching, 97 patients were allocated in each cohort with a mean age of 69.4 and 70.5 years. The two groups showed no difference in operative details except for a higher conversion rate in the NAT group (p = 0.038). The overall postoperative complications rate significantly differed between NAT + surgery (38.1%) and US (21.6%) group (p = 0.019). NAT was found to be related to a higher risk of postoperative morbidity in patients older than 60 years old (p = 0.013) but not in patients younger (p = 0.620). Conversely, no difference in overall survival (p = 0.41) and disease-free-survival (p = 0.34) was found between groups. Conclusions: NAT appears to be related to a higher postoperative complication rate and equivalent oncological outcomes when compared with surgery alone. However, poor short-term outcomes are more evident in patients over 60 years old receiving NAT

CD56, HLA-DR, and CD45 recognize a subtype of childhood AML harboring CBFA2T3-GLIS2 fusion transcript

The presence of CBFA2T3‐GLIS2 fusion gene has been identified in childhood Acute Myeloid Leukemia (AML). In view of the genomic studies indicating a distinct gene expression profile, we evaluated the role of immunophenotyping in characterizing a rare subtype of AML‐CBFA2T3‐GLIS2 rearranged. Immunophenotypic data were obtained by studying a cohort of 20 pediatric CBFA2T3‐GLIS2‐AML and 77 AML patients not carrying the fusion transcript. Enrolled cases were included in the Associazione Italiana di Ematologia Oncologia Pediatrica (AIEOP) AML trials and immunophenotypes were compared using different statistical approaches. By multiple computational procedures, we identified two main core antigens responsible for the identification of the CBFA2T3‐GLIS2‐AML. CD56 showed the highest performance in single marker evaluation (AUC = 0.89) and granted the most accurate prediction when used in combination with HLA‐DR (AUC = 0.97) displaying a 93% sensitivity and 99% specificity. We also observed a weak‐to‐negative CD45 expression, being exceptional in AML. We here provide evidence that the combination of HLA‐DR negativity and intense bright CD56 expression detects a rare and aggressive pediatric AML genetic lesion improving the diagnosis performance

Towards the Composition of Services by End-Users: A Mobile-Based Solution

[EN] Nowadays, we live surrounded by heterogeneous and distributed services that are available to people anytime and anywhere. Even though these services can be used individually, it is through their synchronized and combined usage that end-users are provided with added value. However, existing solutions to service composition are not targeted at ordinary end-users. In fact, these solutions require technical knowledge to deal with the technological heterogeneity in which they are offered to the market. To this end, the paper presents a tool-supported platform that is aided by: (1) EUCalipTool, an end-user mobile tool that implements a Domain Specific Visual Language, which has been specifically designed to compose services on mobile devices; (2) a Faceted Service Registry, which plays the role of gateway between service implementations and end-users, hiding technological issues from the latter when including services in a composition; and (3) a Generation Module, which transforms end-user descriptions into BPMN specification that are interpreted by an execution infrastructure developed for that purpose.This work has been developed with the financial support of the Spanish State Research Agency under the project TIN2017-84094-R and co-financed with ERDF.Valderas, P.; Torres Bosch, MV.; Pelechano Ferragud, V. (2020). Towards the Composition of Services by End-Users: A Mobile-Based Solution. 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A mobile-based solution for supporting end-users in the composition of services

The final publication is available at Springer via http://dx.doi.org/10.1007/s11042-016-3910-4Currently, technologies and applications evolve to create eco-systems made up of a myriad of heterogeneous and distributed services that are accessible anytime and anywhere. Even though these services can be used individually, it is their coordinated and combined usage what provide an added value to end-users. In addition, user¿s wide adoption of mobile devices for daily activities have fostered a shift in the role played by end-users towards Internet data and services. However, existing solutions to service composition are not targeted to ordinary end-users. More easy-to-use tools have to be offered to end-users to make sure that they are successfully accepted and used by them. To this end, the work presented in this paper supports end-users in the creation of service compositions by using mobile devices. We present a Domain Specific Visual Language (DSVL) for end-users that allows them to create service compositions. A tool specifically designed for mobile devices supports this DSVL.This work has been developed with the support of MINECO under the project SMART ADAPT TIN2013-42981-P and co-financed with ERDF.Valderas Aranda, PJ.; Torres Bosch, MV.; Mansanet Benavent, I.; Pelechano Ferragud, V. (2016). A mobile-based solution for supporting end-users in the composition of services. Multimedia Tools and Applications. 1-31. https://doi.org/10.1007/s11042-016-3910-4S131Athreya B, Bahmani F, Diede A, Scaffidi C (2012) End-user programmers on the loose: a study of programming on the phone for the phone. In IEEE Symposium on Visual Languages and Human-Centric Computing (VL/HCC), Innsbruck, Austria, pp. 75–82Atoma (2015) Atoomam, a touch of magic. Accesible at: https://www.atooma.com/ . 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