Optimización del modelo de abordaje a pacientes con asma en Atención Primaria en un entorno COVID-19 / Optimisation of the approach model for asthma patients in Primary Care in a COVID-19 setting

JUSTIFICACIÓN: el asma genera importante morbimortalidad. Una estrategia dirigida a optimizar el modelo de abordaje a pacientes con asma en Atención Primaria en un entorno COVID-19 puede mejorar los resultados en salud.OBJETIVO: crear un marco de actuación de referencia para mejorar el abordaje y satisfacer las necesidades de los pacientes asmáticos en un entorno COVID-19 en Atención Primaria.MATERIAL Y MÉTODOS: se revisó y rediseñó el modelo actual de abordaje de los pacientes adultos con asma en Atención Primaria, desde una perspectiva multidisciplinar, incorporando el concepto Asthma Right Care (1) y el impacto COVID-19.Pensamos que funcionaría por ser un proyecto desarrollado bajo el liderazgo clínico con un enfoque multidisciplinar . Estrategia para el cambio: entre septiembre 2020 - diciembre 2021: Creación de un comité científico. Análisis de la situación actual en 17 centros de salud de Atención Primaria de 7 comunidades autónomas españolas: Personal Médico de Familia, Enfermería y administrativo de cada Centro de Salud con farmacéuticos caracterizaron su modelo de abordaje del paciente asmático e identificaron áreas de mejora, acompañadas de planes de acción. Sesión grupal on line. Informe de resultados para cada centro, con propuesta de indicadores de evaluación de resultado. Laboratorio Digital de Innovación Nacional on line.14-12-2021Usando Mentimeter, referentes de los Centros de Salud y comité científico priorizaron las áreas de mejora identificadas y desarrollaron planes de acción para las dos áreas de mejora más prioritarias. Presentación Informe final del Laboratorio Digital. Impactos esperados• Identificación de áreas de mejora en los itinerarios del paciente asmático en un entorno COVID-19•Diseño participativo de planes de acción seguros y eficientes• Difundir el movimiento Asthma Right Care (AU)

Guía española de la enfermedad pulmonar obstructiva crónica (GesEPOC) 2017: tratamiento farmacológico en fase estable / Spanish guidelines for management of chronic obstructive pulmonary disease (GesEPOC) 2017: pharmacological treatment of stable phase

La enfermedad pulmonar obstructiva crónica (EPOC) presenta una gran heterogeneidad clínica, por lo que su tratamiento se debe individualizar según el nivel de riesgo y el fenotipo. La Guía española de la EPOC (GesEPOC) estableció por primera vez en 2012 unas pautas de tratamiento farmacológico basadas en fenotipos clínicos. Estas pautas han sido adoptadas posteriormente por otras normativas nacionales, y han sido respaldadas por nuevas evidencias publicadas desde entonces. En esta actualización 2017 se ha sustituido la clasificación de gravedad inicial por una clasificación de riesgo mucho más sencilla (bajo o alto riesgo), basándose en la función pulmonar, el grado de disnea y la historia de agudizaciones, y se recomienda la determinación del fenotipo clínico únicamente en pacientes de alto riesgo. Se mantienen los mismos fenotipos clínicos: no agudizador, EPOC-asma (ACO), agudizador con enfisema y agudizador con bronquitis crónica. La base del tratamiento farmacológico de la EPOC es la broncodilatación, y también es el único tratamiento recomendado en pacientes de bajo riesgo. En los pacientes con alto riesgo se añadirán diversos fármacos a los broncodilatadores según el fenotipo clínico. GesEPOC supone una aproximación al tratamiento de la EPOC más individualizado según las características clínicas de los pacientes y su nivel de riesgo o de complejidad.(AU)

The clinical presentation of chronic obstructive pulmonary disease (COPD) varies widely, so treatment must be tailored according to the level of risk and phenotype. In 2012, the Spanish COPD Guidelines (GesEPOC) first established pharmacological treatment regimens based on clinical phenotypes. These regimens were subsequently adopted by other national guidelines, and since then, have been backed up by new evidence. In this 2017 update, the original severity classification has been replaced by a much simpler risk classification (low or high risk), on the basis of lung function, dyspnea grade, and history of exacerbations, while determination of clinical phenotype is recommended only in high-risk patients. The same clinical phenotypes have been maintained: non-exacerbator, asthma-COPD overlap (ACO), exacerbator with emphysema, and exacerbator with bronchitis. Pharmacological treatment of COPD is based on bronchodilators, the only treatment recommended in low-risk patients. High-risk patients will receive different drugs in addition to bronchodilators, depending on their clinical phenotype. GesEPOC reflects a more individualized approach to COPD treatment, according to patient clinical characteristics and level of risk or complexity.(AU)

Development and applications of a DNA labeling method with magnetic nanoparticles to study the role of horizontal gene transfer events between bacteria in soil pollutant bioremediation processes.

Horizontal gene transfers are critical mechanisms of bacterial evolution and adaptation that are involved to a significant level in the degradation of toxic molecules such as xenobiotic pesticides. However, understanding how these mechanisms are regulated in situ and how they could be used by man to increase the degradation potential of soil microbes is compromised by conceptual and technical limitations. This includes the physical and chemical complexity and heterogeneity in such environments leading to an extreme bacterial taxonomical diversity and a strong redundancy of genes and functions. In addition, more than 99 % of soil bacteria fail to develop colonies in vitro, and even new DNA-based investigation methods (metagenomics) are not specific and sensitive enough to consider lysis recalcitrant bacteria and those belonging to the rare biosphere. The objective of the ANR funded project "Emergent" was to develop a new culture independent approach to monitor gene transfer among soil bacteria by labeling plasmid DNA with magnetic nanoparticles in order to specifically capture and isolate recombinant cells using magnetic microfluidic devices. We showed the feasibility of the approach by using electrotransformation to transform a suspension of Escherichia coli cells with biotin-functionalized plasmid DNA molecules linked to streptavidin-coated superparamagnetic nanoparticles. Our results have demonstrated that magnetically labeled cells could be specifically retained on micromagnets integrated in a microfluidic channel and that an efficient selective separation can be achieved with the microfluidic device. Altogether, the project offers a promising alternative to traditional culture-based approaches for deciphering the extent of horizontal gene transfer events mediated by electro or natural genetic transformation mechanisms in complex environments such as soil.

Inter-laboratory evaluation of the ISO standard 11063 "Soil quality - Method to directly extract DNA from soil samples".

Extracting DNA directly from micro-organisms living in soil is a crucial step for the molecular analysis of soil microbial communities. However, the use of a plethora of different soil DNA extraction protocols, each with its own bias, makes accurate data comparison difficult. To overcome this problem, a method for soil DNA extraction was proposed to the International Organization for Standardization (ISO) in 2006. This method was evaluated by 13 independent European laboratories actively participating in national and international ring tests. The reproducibility of the standardized method for molecular analyses was evaluated by comparing the amount of DNA extracted, as well as the abundance and genetic structure of the total bacterial community in the DNA extracted from 12 different soils by the 13 laboratories. High quality DNA was successfully extracted from all 12 soils, despite different physical and chemical characteristics and a range of origins from arable soils, through forests to industrial sites. Quantification of the 16S rRNA gene abundances by real time PCR and analysis of the total bacterial community structure by automated ribosomal intergenic spacer analysis (A-RISA) showed acceptable to good levels of reproducibility. Based on the results of both ring-tests, the method was unanimously approved by the ISO as an international standard method and the normative protocol will now be disseminated within the scientific community. Standardization of a soil DNA extraction method will improve data comparison, facilitating our understanding of soil microbial diversity and soil quality monitoring.

[Polylactic acid injections (Newfill) in the treatment of facial lipodystrophy in HIV-positive patients]. / Traitement par injections d'acide polylactique (Newfill) des lipoatrophies faciales chez les malades infectés par le VIH.

BACKGROUND: Lipodystrophy is a side-effect associated with treatment for human immunodeficiency virus (HIV) and is found chiefly on the face (disappearance of buccal fat pads) and is detrimental to self-esteem. PATIENTS AND METHODS: This was a retrospective study in HIV-positive patients with facial lipoatrophy treated between 1999 and 2004 by means of subcutaneous injections of polylactic acid (Newfill). We assessed the efficacy of treatment, the number of injections given, treatment methods and adverse effects. RESULTS: Eighty-three patients were treated between 1999 and 2004. Each patient received a mean of between 3 and 4 treatment sessions comprising subdermal injection of 1 ampoule of Newfill into each cheek. Ultrasound assessment of the dermis over the cheekbone was performed in 45 patients and showed an increase in dermal thickness of between 3 and 7 mm. Following injection, edema was observed in all cases and lasted between 1 and 2 days. Five patients presented bruising at the injection sites. Two patients presented asymmetry lasting 4 months and requiring correction. Four patients had non-inflammatory granulomas, which were not visible but were palpable in 2 cases; all nodules regressed after 4 months. DISCUSSION: Treatment of facial lipoatrophy in HIV patients by injection of polylactic acid (Newfill) was shown to be efficacious in the majority of subjects. Training in the administration of this treatment is needed to ensure optimal efficacy and safety. A number of technical difficulties led to changes in treatment methods, i.e. routine adoption of a mask and protective glasses and use of a Luer-lock syringe due to blockage of syringes in more than 20% of cases, with splashing; increase in dilution volume from 3 to 5 ml; use of a centrifuge to ensure greater homogeneity of the solution; use of lidocaine in place of water for injections in order to reduce pain for patients.

Variable virological outcomes according to the center providing antiretroviral therapy within the PharmAdapt clinical trial.

PURPOSE: Differences in virological response between HIV-infected patients at different study centers were analyzed as a substudy of PharmAdapt, a multicenter prospective randomized study to evaluate the utility of therapeutic drug monitoring after a genotypic-based treatment adaptation. RESULTS: After 12 weeks, the percentage of patients participating in PharmAdapt with HIV RNA < 200 copies/mL ranged from 17% to 69% between centers providing antiretroviral care. In a multivariate analysis, independent factors predictive of viral load <200 HIV RNA copies/mL at Week 12 included: lower baseline viral load, lower nonnucleoside reverse transcriptase inhibitor resistance, lower protease inhibitor resistance, and the center providing antiretroviral therapy. To evaluate the final factor, study sites were divided into two groups based on Week 12 HIV RNA values above or below the median. CONCLUSION: Using this definition, observed differences between centers included the use of stavudine, abacavir-, and/or efavirenz-based regimens and use of online expert advice.

Opportunistic colonization of Ralstonia solanacearum-infected plants by Acinetobacter sp. and its natural competence development.

The behavior of the soil bacterium Acinetobacter sp. BD413 was monitored in Ralstonia solanacearum-infected and non-infected tomato plants after direct injection into the stem or natural infection by roots. In healthy plants, Acinetobacter sp. BD413 failed to colonize plant tissue. In plants infected simultaneously by the pathogen R. solanacearum,the Acinetobacter population increased linearly to about 3.1 x 10(7) cells per gram plant material and was maintained at a high level until the death of the plant. Moreover, Acinetobacter sp. BD413 was found to develop a competent state when multiplying in planta, indicating it could possibly be transformed by bacterial or plant DNA.

Quantification of bacterial subgroups in soil: comparison of DNA extracted directly from soil or from cells previously released by density gradient centrifugation.

All molecular analyses of soil bacterial diversity are based on the extraction of a representative fraction of cellular DNA. Methods of DNA extraction for this purpose are divided into two categories: those in which cells are lysed within the soil (direct extraction) and those in which cells are first removed from soil (cell extraction) and then lysed. The purpose of this study was to compare a method of direct extraction with a method in which cells were first separated from the soil matrix by Nycodenz gradient centrifugation in order to evaluate the effect of these different approaches on the analysis of the spectrum of diversity in a microbial community. We used a method based on polymerase chain reaction (PCR) amplification of a 16S rRNA gene fragment, followed by hybridization of the amplified fragments to a set of specific probes to assess the phylogenetic diversity of our samples. Control parameters, such as the relationship between amount of DNA template and amount of PCR product and the influence of competing DNA on PCR amplification, were first examined. Comparison between extraction methods showed that less DNA was extracted when cells were first separated from the soil matrix (0.4 microg g(-1) dry weight soil versus 38-93 microg g(-1) obtained by in situ lysis methods). However, with the exception of the gamma-subclass of Proteobacteria, there was no significant difference in the spectrum of diversity resulting from the two extraction strategies.

Resistance of plastic ophthalmic lenses: the effect of base curve on different materials during static load testing.

PURPOSE: This study was designated to evaluate, through a static load test, the influence of lens base curve on the fracture resistance of three common plastic materials. METHODS: A JJ Lloyd load cell machine was used to test the fracture resistance of -4.00 D spherical lenses. The samples had a nominal center thickness of 2.0 mm and a base curve distributed in one of five groups (+0.50, +2.50, +4.50, +6.50, and +8.50 D). The lenses were manufactured in CR39, polycarbonate, and TL16, a high refractive index plastic (n = 1.599). RESULTS: The lens base curve influenced fracture resistance for all materials. For these materials, resistance increased as the base curve varied from +0.50 to +8.50 D. The resistance of CR39, TL16, and polycarbonate lenses was found to be linearly dependent on lens base curve. The effect is stronger for polycarbonate. Fracture resistance was higher for TL16 than for CR39, and polycarbonate was much more resistant to breakage than the two other materials. CONCLUSIONS: For a given power, the fracture resistance of an ophthalmic lens is reduced when its base curve has a low value. Consequently, the flattening of ophthalmic lenses for cosmetic purposes is not recommended as far as fracture resistance is concerned.

Laboratory-scale evidence for lightning-mediated gene transfer in soil.

Electrical fields and current can permeabilize bacterial membranes, allowing for the penetration of naked DNA. Given that the environment is subjected to regular thunderstorms and lightning discharges that induce enormous electrical perturbations, the possibility of natural electrotransformation of bacteria was investigated. We demonstrated with soil microcosm experiments that the transformation of added bacteria could be increased locally via lightning-mediated current injection. The incorporation of three genes coding for antibiotic resistance (plasmid pBR328) into the Escherichia coli strain DH10B recipient previously added to soil was observed only after the soil had been subjected to laboratory-scale lightning. Laboratory-scale lightning had an electrical field gradient (700 versus 600 kV m(-1)) and current density (2.5 versus 12.6 kA m(-2)) similar to those of full-scale lightning. Controls handled identically except for not being subjected to lightning produced no detectable antibiotic-resistant clones. In addition, simulated storm cloud electrical fields (in the absence of current) did not produce detectable clones (transformation detection limit, 10(-9)). Natural electrotransformation might be a mechanism involved in bacterial evolution.

Recomendaciones para la atención al paciente con enfermedad pulmonar obstructiva crónica / Guidelines for care of the patient with chronic obstructive pulmonary disease

No disponible

Natural transformation of Pseudomonas fluorescens and Agrobacterium tumefaciens in soil.

Little information is available concerning the occurrence of natural transformation of bacteria in soil, the frequency of such events, and the actual role of this process on bacterial evolution. This is because few bacteria are known to possess the genes required to develop competence and because the tested bacteria are unable to reach this physiological state in situ. In this study we found that two soil bacteria, Agrobacterium tumefaciens and Pseudomonas fluorescens, can undergo transformation in soil microcosms without any specific physical or chemical treatment. Moreover, P. fluorescens produced transformants in both sterile and nonsterile soil microcosms but failed to do so in the various in vitro conditions we tested. A. tumefaciens could be transformed in vitro and in sterile soil samples. These results indicate that the number of transformable bacteria could be higher than previously thought and that these bacteria could find the conditions necessary for uptake of extracellular DNA in soil.

Evaluation of biological and physical protection against nuclease degradation of clay-bound plasmid DNA.

In order to determine the mechanisms involved in the persistence of extracellular DNA in soils and to monitor whether bacterial transformation could occur in such an environment, we developed artificial models composed of plasmid DNA adsorbed on clay particles. We determined that clay-bound DNA submitted to an increasing range of nuclease concentrations was physically protected. The protection mechanism was mainly related to the adsorption of the nuclease on the clay mineral. The biological potential of the resulting DNA was monitored by transforming the naturally competent proteobacterium Acinetobacter sp. strain BD413, allowing us to demonstrate that adsorbed DNA was only partially available for transformation. This part of the clay-bound DNA which was available for bacteria, was also accessible to nucleases, while the remaining fraction escaped both transformation and degradation. Finally, transformation efficiency was related to the perpetuation mechanism, with homologous recombination being less sensitive to nucleases than autonomous replication, which requires intact molecules.

Transverse chromatic aberration and colour-defined motion.

A number of recent studies have explored the role of the chromatic system in motion processes using the isoluminance paradigm. A major concern when using such methodological procedures is potential artefacts produced by chromatic aberrations. In the present study we address the problem of optically induced luminance artefacts produced by transverse chromatic aberrations (TCA), which may contaminate the results obtained in chromatic motion-nulling experiments. Results show that different TCA levels artificially increase chromatic motion sensitivity values to varying degrees above 0.5 cpd for red/green gratings. The data also suggest the notion that naturally occurring TCA can decrease motion-nulling thresholds for chromatic gratings at high spatial frequencies. Furthermore, our data show that the motion-nulling paradigm for chromatic gratings may in fact be an efficient functional method for assessing the amount of TCA produced by optical factors.

Plant genome complexity may be a factor limiting in situ the transfer of transgenic plant genes to the phytopathogen Ralstonia solanacearum.

The development of natural competence by bacteria in situ is considered one of the main factors limiting transformation-mediated gene exchanges in the environment. Ralstonia solanacearum is a plant pathogen that is also a naturally transformable bacterium that can develop the competence state during infection of its host. We have attempted to determine whether this bacterium could become the recipient of plant genes. We initially demonstrated that plant DNA was released close to the infecting bacteria. We constructed and tested various combinations of transgenic plants and recipient bacteria to show that the effectiveness of such transfers was directly related to the ratio of the complexity of the plant genome to the number of copies of the transgene.

Potential dissemination of antibiotic resistance genes from transgenic plants to microorganisms.

Evidence that genes were transferred during evolution from plants to bacteria was obtained from nucleotide and protein sequence analyses. However, the extent of such transfers among phylogenetically distant organisms is limited by various factors, including those related to complexity of the environment and those endogenous to the bacteria, designed to prevent a drift of the genome integrity. The goal of this article is to give an overview of the potentials and limits of natural interkingdom gene transfers, with a particular focus on prokaryote originating sequences fitting the nuclear genome of transgenic plants.

Comparison between electrochemical and optoelectrochemical impedance measurements for detection of DNA hybridization.

The principles of the electrochemical and optoelectrochemical impedance measurements on bare electrolyte/dielectric/semiconductor structures are described. The analysis of the experimental curves allows access to several indications concerning the electrical behavior of such structures. The application of these techniques to follow the electrical behavior of structures modified with two biological systems was investigated. The antibody/antigen recognition did not change the surface charge and, therefore, did not affect the impedance curves with respect to the applied potential. By contrast, the hybridization of two complementary DNA strands on the surface of the structure induced a variation of flat band potential of the semiconductor leading to a shift of impedance curves along the potential axis. This means that it is possible to detect directly the DNA hybridization without the use of labeled probes. The use of light allows the surface to be probed locally. In the future, the application of this technique for direct detection of hybridization on DNA chips should be possible.

Quantification of bias related to the extraction of DNA directly from soils.

In recent years, several protocols based on the extraction of nucleic acids directly from the soil matrix after lysis treatment have been developed for the detection of microorganisms in soil. Extraction efficiency has often been evaluated based on the recovery of a specific gene sequence from an organism inoculated into the soil. The aim of the present investigation was to improve the extraction, purification, and quantification of DNA derived from as large a portion of the soil microbial community as possible, with special emphasis placed on obtaining DNA from gram-positive bacteria, which form structures that are difficult to disrupt. Furthermore, we wanted to identify and minimize the biases related to each step in the procedure. Six soils, covering a range of pHs, clay contents, and organic matter contents, were studied. Lysis was carried out by soil grinding, sonication, thermal shocks, and chemical treatments. DNA was extracted from the indigenous microflora as well as from inoculated bacterial cells, spores, and hyphae, and the quality and quantity of the DNA were determined by gel electrophoresis and dot blot hybridization. Lysis efficiency was also estimated by microscopy and viable cell counts. Grinding increased the extracellular DNA yield compared with the yield obtained without any lysis treatment, but none of the subsequent treatments clearly increased the DNA yield. Phage lambda DNA was inoculated into the soils to mimic the fate of extracellular DNA. No more than 6% of this DNA could be recovered from the different soils. The clay content strongly influenced the recovery of DNA. The adsorption of DNA to clay particles decreased when the soil was pretreated with RNA in order to saturate the adsorption sites. We also investigated different purification techniques and optimized the PCR methods in order to develop a protocol based on hybridization of the PCR products and quantification by phosphorimaging.

Horizontal gene transfers in the environment: natural transformation as a putative process for gene transfers between transgenic plants and microorganisms.

Horizontal gene transfers among bacteria, such as natural transformation or conjugation, may have played an important role in bacterial evolution. They are thought to have been involved in promoting genome plasticity which permitted bacteria to adapt very efficiently to any change in their environment and to colonize a wide range of ecosystems. Evidence that some genes were transferred from eukaryotes, and in particular, from plants to bacteria, was obtained from nucleotide and protein sequence analyses. However, numerous factors, including some which are endogenous to the bacterial cells, tend to limit the extent of transfer, particularly among phylogenetically distant organisms. The goal of this paper is to give an overview of the potentials and limits of natural interkingdom gene transfers, with particular focus on prokaryote-originating sequences which fit the nuclear genome of transgenic plants.