Occurrence of the p019 Gene in the blaKPC-Harboring Plasmids: Adverse Clinical Impact for Direct Tracking of KPC-Producing Klebsiella pneumoniae by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been used for the direct detection of KPC-producing isolates by analysis of the 11,109 Da mass peak representing the P019 protein. In this study, we evaluate the presence of the 11,109 Da mass peak in a collection of 435 unduplicated Klebsiella pneumoniae clinical isolates. The prevalence of the P019 peak in the blaKPC K. pneumoniae isolates was 49.2% (32/65). The 11,109 Da mass peak was not observed in any of the other carbapenemase (319) or noncarbapenemase producers (116). Computational analysis of the presence of the p019 gene was performed in the aforementioned carbapenemase-producing K. pneumoniae isolates fully characterized by whole-genome sequencing (WGS) and in a further collection of 1,649 K. pneumoniae genomes included in EuSCAPE. Herein, we have demonstrated that the p019 gene is not exclusively linked to the pKpQil plasmid but that it is present in the following plasmids: IncFIB(K)/IncFII(K)/ColRNAI, IncFIB(pQil), IncFIB(pQil)/ColRNAI, IncFIB(pQil)/IncFII(K), IncFIB(K)/IncFII(K), and IncX3. In addition, we have proven the independent movement of the Tn4401 and the ISKpn31, of which the p019 gene is a component. The absence of the p019 gene was obvious in Col440I, Col(pHAD28), IncFIB(K)/IncX3/IncFII(K), and IncFIB(K)/IncFII(K) plasmids. In addition, we also observed another plasmid in which neither Tn4401 nor ISKpn31 was found, IncP6. In the EuSCAPE, the occurrence of p019 varied from 0% to 100% among the different geographical locations. The adverse clinical impact of the diminished prevalence of the p019 gene within the plasmid encoding KPC-producing Klebsiella pneumoniae puts forward the need for reconsideration when applying this technique in a clinical setting.

An Improved Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Data Analysis Pipeline for the Identification of Carbapenemase-Producing Klebsiella pneumoniae.

The increasing emergence of carbapenemase-producing Klebsiella pneumoniae (CPK) isolates is a global health alarm. Rapid methods that require minimum sample preparation and rapid data analysis are urgently required. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been used by clinical laboratories for identification of antibiotic-resistant bacteria; however, discrepancies have arisen regarding biological and technical issues. The aim of this study was to standardize an operating procedure and data analysis for identification of CPK by MALDI-TOF MS. To evaluate this approach, a series of 162 K. pneumoniae isolates (112 CPK and 50 non-CPK) were processed in the MALDI BioTyper system (Bruker Daltonik, Germany) following a standard operating procedure. The study was conducted in two stages; the first is denominated the "reproducibility stage" and the second "CPK identification." The first stage was designed to evaluate the biological and technical variation associated with the entire analysis of CPK and the second stage to assess the final accuracy of MALDI-TOF MS for the identification of CPK. Therefore, we present an improved MALDI-TOF MS data analysis pipeline using neural network analysis implemented in Clover MS Data Analysis Software (Clover Biosoft, Spain) that is designed to reduce variability, guarantee interlaboratory reproducibility, and maximize the information selected from the bacterial proteome. Using the random forest (RF) algorithm, 100% of CPK isolates were correctly identified when all the peaks in the spectra were selected as input features and total ion current (TIC) normalization was applied. Thus, we have demonstrated that real-time direct tracking of CPK is possible using MALDI-TOF MS.

Estimación muestral en la percepción de los servicios deportivos a través de la teoría de la generalizabilidad / Estimation of simple in the perception of the sports services through the generalizability theory

La presente investigación utiliza la Teoría de la Generalizabilidad (TG) aplicada al ámbito de la gestión deportiva con el objetivo de conocer el tamaño muestral adecuado para la evaluación de la calidad percibida. Para ello, se utilizó una estructura compuesta por las facetas ítems (I) y participantes (P), con un diseño totalmente cruzado. Dentro del Modelo Lineal General (GLM), con estas dos facetas, se realizó un análisis de la varianza y se estimó la precisión de generalización. La herramienta empleada fue un cuestionario compuesto por 25 ítems y los participantes, un total de 330, pertenecían a una instalación deportiva de la ciudad de Málaga. Los resultados muestran que el número de participantes utilizado obtuvo un registro fiable (ξρ²(δ) = 0.98), y permitió estimar el nivel de precisión en la generalización para el tamaño muestral empleado (coeficiente de generalizabilidad, ξρ²(Δ) = 0.96), suponiendo de esta manera un importante avance en cuanto a la eficiencia en futuras evaluaciones de servicios deportivos

The present study uses the Generalizability Theory (GT) apply into the sport management area aiming to know the accurate sample size to evaluate the perceived quality. For this purpose, a compound structure was used for the participants (P) and items (I) aspects, with a fully crossed design. Applying general linear model (GLM), with both aspects, a variance analysis was realised and it estimated the generalization's precision. The instrument was a survey composed for 25 items and the sample, in total 330, comes from a fitness centre in the city of Malaga. The obtain results shows a number of participants using a reliable register (ξρ²(δ) = 0.98), and it estimates the precision level in the generalization to the sample size used (generalizability coefficient, ξρ²(Δ) = 0.96), assuming in this side an important progress in relation to the efficiency in upcoming evaluations in sports services

The relationship among service experience, perceived value and behavioural intentions of customers in a group fitness class / La relación entre la experiencia de servicio, el valor percibido y las intenciones de comportamiento en clientes de actividades dirigidas

Understanding the importance of the customer perception during the experience in the fitness centre is fundamental to comprehend behavioural intentions for fitness centre managers. This study attempts to understand how important and necessary is consumer’s assessment of service experience, perceived value, and behavioural intention, and the relationships among theses variables into group fitness class (GFC), and Virtual fitness class (VFC). Data was collected from 1.943 customers. The confirmatory factor analysis indicated an adequate goodness-of-fit-indexes for the model obtained. Structural equation modeling analysis revealed the positive and significant effect of service experience on customers’ perceived value and behavioural intention, but results show that perceived value has poor fit on behavioural intentions. The overall customer’s perception of GFC and VFC can be a focus for fitness centre’s managers aiming to improve fitness service supply related to new technologies in the fitness industry to increase intentions to repurchase

Comprender la percepción del consumidor durante su experiencia dentro de un centro de fitness resulta fundamental para que los gerentes puedan entender las intenciones de comportamiento futuras de sus abonados. Este estudio pretende esclarecer la importancia y la necesaria evaluación de la experiencia de servicio, el valor percibido y la intenciones de comportamiento, así como la relación entre dichas variables dentro de las actividades dirigidas con técnico y las actividades dirigidas virtuales, utilizando para ello una muestra de 1.943 usuarios. El análisis factorial confirmatorio indica un adecuado ajuste para el modelo de medida propuesto. Por otro lado, el análisis de ecuaciones estructurales utilizado revela una relación positiva y significativa de las experiencias de servicio hacia el valor percibido y las intenciones de comportamiento respectivamente. Sin embargo, el modelo muestra un pobre ajuste entre el valor percibido y las intenciones de comportamiento. La percepción global del consumidor en ambas actividades puede resultar un elemento de estudio para los gerentes con el objetivo de mejorar la oferta de servicios de fitness utilizando las nuevas tecnologías para así incrementar las intenciones de compra