Inactivation of the thymidylate synthase thyA in non-typeable Haemophilus influenzae modulates antibiotic resistance and has a strong impact on its interplay with the host airways.

Antibacterial treatment with cotrimoxazol (TxS), a combination of trimethoprim and sulfamethoxazole, generates resistance by, among others, acquisition of thymidine auxotrophy associated with mutations in the thymidylate synthase gene thyA, which can modify the biology of infection. The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) is frequently encountered in the lower airways of chronic obstructive pulmonary disease (COPD) patients, and associated with acute exacerbation of COPD symptoms. Increasing resistance of NTHi to TxS limits its suitability as initial antibacterial against COPD exacerbation, although its relationship with thymidine auxotrophy is unknown. In this study, the analysis of 2,542 NTHi isolates recovered at Bellvitge University Hospital (Spain) in the period 2010-2014 revealed 119 strains forming slow-growing colonies on the thymidine low concentration medium Mueller Hinton Fastidious, including one strain isolated from a COPD patient undergoing TxS therapy that was a reversible thymidine auxotroph. To assess the impact of thymidine auxotrophy in the NTHi-host interplay during respiratory infection, thyA mutants were generated in both the clinical isolate NTHi375 and the reference strain RdKW20. Inactivation of the thyA gene increased TxS resistance, but also promoted morphological changes consistent with elongation and impaired bacterial division, which altered H. influenzae self-aggregation, phosphorylcholine level, C3b deposition, and airway epithelial infection patterns. Availability of external thymidine contributed to overcome such auxotrophy and TxS effect, potentially facilitated by the nucleoside transporter nupC. Although, thyA inactivation resulted in bacterial attenuation in a lung infection mouse model, it also rendered a lower clearance upon a TxS challenge in vivo. Thus, our results show that thymidine auxotrophy modulates both the NTHi host airway interplay and antibiotic resistance, which should be considered at the clinical setting for the consequences of TxS administration

Escribir en la universidad: aplicación de nuevas tecnologías

Depto. de Lengua Española y Teoría de la LiteraturaFac. de FilologíaFALSEsubmitte

Study protocol of a randomized controlled trial to assess safety of teleconsultation compared with face-to-face consultation: the ECASeT study

BackgroundThe use of remote consultation modalities has exponentially grown in the past few years, particularly since the onset of the COVID-19 pandemic. Although a huge body of the literature has described the use of phone (tele) and video consultations, very few of the studies correspond to randomized controlled trials, and none of them has assessed the safety of these consultation modalities as the primary objective. The primary objective of this trial was to assess the safety of remote consultations (both video and teleconsultation) in the follow-up of patients in the hospital setting.MethodsMulticenter, randomized controlled trial being conducted in four centers of an administrative healthcare area in Catalonia (North-East Spain). Participants will be screened from all individuals, irrespective of age and sex, who require follow-up in outpatient consultations of any of the departments involved in the study. Eligibility criteria have been established based on the local guidelines for screening patients for remote consultation. Participants will be randomly allocated into one of the two study arms: conventional face-to-face consultation (control) and remote consultation, either teleconsultation or video consultation (intervention). Routine follow-up visits will be scheduled at a frequency determined by the physician based on the diagnostic and therapy of the baseline disease (the one triggering enrollment). The primary outcome will be the number of adverse reactions and complications related to the baseline disease. Secondary outcomes will include non-scheduled visits and hospitalizations, as well as usability features of remote consultations. All data will either be recorded in an electronic clinical report form or retrieved from local electronic health records. Based on the complications and adverse reaction rates reported in the literature, we established a target sample size of 1068 participants per arm. Recruitment started in May 2022 and is expected to end in May 2024.DiscussionThe scarcity of precedents on the assessment of remote consultation modalities using randomized controlled designs challenges making design decisions, including recruitment, selection criteria, and outcome definition, which are discussed in the manuscript.Trial registrationNCT05094180. The items of the WHO checklist for trial registration are available in Additional file 1. Registered on 24 November 2021

La inactivación de la timidilato sintetasa ThyA modula la resistencia antibiótica y la interacción de Haemophilus influenzae con el aparato respiratorio humano

Póster presentado en la XI Reunión del Grupo de Microbiología Molecular de la Sociedad Española de Microbiología (SEM), celebrada en Sevilla del 6 al 8 de septiembre de 2016.Haemophilus influenzae (Hi) es un miembro habitual del microbioma respiratorio y el patógeno oportunista más frecuentemente aislado en muestras de esputo recogidas durante la exacerbación de pacientes con Enfermedad Pulmonar Obstructiva Crónica (EPOC). El tratamiento de las exacerbaciones implica la administración de antibióticos como Cotrimoxazol (SMX), y la consiguiente adquisición de resistencias. Entre los mecanismos de resistencia a SMX, se encuentra la auxotrofía a timidina asociada a mutaciones en el gen thyA, que codifica una timidilato sintetasa. En un estudio observacional de 2,542 aislados clínicos EPOC de Hi obtenidos entre 2010 y 2014 en el HU Bellvitge (Barcelona), 119 formaron colonias pequeñas con crecimiento lento en placas de Mueller Hinton Fastidioso (MHF), un medio con baja concentración de timidina. El análisis de estos 119 aislados identificó Hi8233, una cepa de un paciente que recibió SMX, auxótrofa natural a timidina, resistente a SMX (SMXR), con una inserción de seis nucleótidos en thyA. Para entender la relación entre auxotrofía a timidina, SMXR e interacción hospedador-patógeno durante la infección respiratoria por Hi, se generaron y caracterizaron los mutantes HiRdKW20ΔthyA y Hi375ΔthyA, en relación a auxotrofía a timidina, SMXR, morfología, crecimiento, auto-agregación, expresión génica, interacción con inmunidad innata, e infección pulmonar. La inactivación del gen thyA genera cepas Hi SMXR y produce auxotrofía a timidina. Esta auxotrofía se revierte en placas de MHF y cultivo líquido por adición de timidina exógena, y genera cambios morfológicos asociados a menor tasa de división, lo que aumenta el tamaño y facilita la auto-agregación bacteriana. La expresión del transportador de nucleósidos nupC aumenta en cepas HiΔthyA, lo que sugiere que la reversión fenotípica por adición de timidina exógena está relacionada con su captación. Asimismo, la auxotrofía a timidina disminuye la adhesióninvasión epitelial, y la carga bacteriana en pulmón y lavado broncoalveolar murino, si bien aumenta el fitness del patógeno en animales sometidos a un régimen terapéutico consistente en la administración oral de SMX. En conjunto, la auxotrofía a timidina en Hi es un mecanismo de SMXR que, concomitantemente, disminuye su capacidad infectiva. Este equilibrio resistencia antibiótica-virulencia debe ser clínicamente considerado en el tratamiento de las exacerbaciones EPOC.N

El desbalance del metabolismo de glucosa por inactivación génica condiciona la interacción hospedador-patógeno durante la infección respiratoria por Haemophilus invluenzae

Trabajo presentado en la XII Reunión del Grupo Especializado SEM, celebrada en Zaragoza (España) del 5 al 7 de septiembre de 2018Peer reviewe

Application of a semiautomatic classifier for modic and disk hernia changes in magnetic resonance

OBJECTIVE: Early detection of degenerative changes in lumbar intervertebral disc by magnetic resonance imaging in a semiautomatic classifier for prevention of degenerative disease. METHOD: MRIs were selected with a diagnosis of degenerative disc disease or back pain from January to May 2014, with a sample of 23 patients and a total of 170 disks evaluated by sagittal T2 MRI image, first evaluated by a specialist physician in training and them were introduced into the software, being the results compared. RESULTS: One hundred and fifteen discs were evaluated by a programmed semiautomatic classifier to identify MODIC changes and hernia, which produced results "normal or MODIC" and "normal or abnormal", respectively. With a total of 230 readings, of which 141 were correct, 84 were reading errors and 10 readings were undiagnosed, the semiautomatic classifier is a useful tool for early diagnosis or established disease and is easy to apply because of the speed and ease of use; however, at this early stage of development, software is inferior to clinical observations and the results were from around 65% to 60% certainty for MODIC rating and 61% to 58% for disc herniation, compared with clinical evaluations. CONCLUSION: The comparative results between the two doctors were 94 consistent results and only 21 errors, which represents 81% certainty

Inactivation of the Thymidylate Synthase thyA in Non-typeable Haemophilus influenzae Modulates Antibiotic Resistance and Has a Strong Impact on Its Interplay with the Host Airways

Antibacterial treatment with cotrimoxazol (TxS), a combination of trimethoprim and sulfamethoxazole, generates resistance by, among others, acquisition of thymidine auxotrophy associated with mutations in the thymidylate synthase gene thyA, which can modify the biology of infection. The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) is frequently encountered in the lower airways of chronic obstructive pulmonary disease (COPD) patients, and associated with acute exacerbation of COPD symptoms. Increasing resistance of NTHi to TxS limits its suitability as initial antibacterial against COPD exacerbation, although its relationship with thymidine auxotrophy is unknown. In this study, the analysis of 2,542 NTHi isolates recovered at Bellvitge University Hospital (Spain) in the period 2010–2014 revealed 119 strains forming slow-growing colonies on the thymidine low concentration medium Mueller Hinton Fastidious, including one strain isolated from a COPD patient undergoing TxS therapy that was a reversible thymidine auxotroph. To assess the impact of thymidine auxotrophy in the NTHi-host interplay during respiratory infection, thyA mutants were generated in both the clinical isolate NTHi375 and the reference strain RdKW20. Inactivation of the thyA gene increased TxS resistance, but also promoted morphological changes consistent with elongation and impaired bacterial division, which altered H. influenzae self-aggregation, phosphorylcholine level, C3b deposition, and airway epithelial infection patterns. Availability of external thymidine contributed to overcome such auxotrophy and TxS effect, potentially facilitated by the nucleoside transporter nupC. Although, thyA inactivation resulted in bacterial attenuation in a lung infection mouse model, it also rendered a lower clearance upon a TxS challenge in vivo. Thus, our results show that thymidine auxotrophy modulates both the NTHi host airway interplay and antibiotic resistance, which should be considered at the clinical setting for the consequences of TxS administration.IR is funded by a Ph.D. studentship from Universidad Pública de Navarra, Spain; JM is funded by Ph.D. studentship BES-2013-062644 from Ministerio Economía y Competitividad-MINECO, Spain; SM is funded by a postdoctoral contract from CIBER Enfermedades Respiratorias (CIBERES); NL is funded by a contract from Department of Economy, Regional Govern from Navarra, Spain, reference 0011-1307-2015-000037. This work has been funded by grants from MINECO SAF2012-31166 and SAF2015-66520-R, Health Department, Regional Govern from Navarra, Spain, reference 03/2016, and SEPAR 31/2015 to JG. CIBERES is an initiative from Instituto de Salud Carlos III (ISCIII), Madrid, Spain. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).Peer reviewe

Guía de aprovechamiento de recursos didácticos : matemáticas : primer ciclo de la ESO

Ejemplar fotocopiadoGuía que proporciona información sobre los recursos didácticos del área de matemáticos para el primer ciclo de la ESO. Se indican los criterios de selección de los materiales didácticos. Se subdividen por tipos de recursos; bibliográficos, audiovisuales e informáticos y materiales. Los recursos se presentan con la descripción bibliográfica y el resumen.MadridBiblioteca de Educación del Ministerio de Educación, Cultura y Deporte; Calle San Agustín 5 -3 Planta; 28014 Madrid; Tel. +34917748000; [email protected]

Inactivation of the thymidylate synthase thyA in non-typeable Haemophilus influenzae modulates antibiotic resistance and has a strong impact on its interplay with the host airways.

Antibacterial treatment with cotrimoxazol (TxS), a combination of trimethoprim and sulfamethoxazole, generates resistance by, among others, acquisition of thymidine auxotrophy associated with mutations in the thymidylate synthase gene thyA, which can modify the biology of infection. The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) is frequently encountered in the lower airways of chronic obstructive pulmonary disease (COPD) patients, and associated with acute exacerbation of COPD symptoms. Increasing resistance of NTHi to TxS limits its suitability as initial antibacterial against COPD exacerbation, although its relationship with thymidine auxotrophy is unknown. In this study, the analysis of 2,542 NTHi isolates recovered at Bellvitge University Hospital (Spain) in the period 2010-2014 revealed 119 strains forming slow-growing colonies on the thymidine low concentration medium Mueller Hinton Fastidious, including one strain isolated from a COPD patient undergoing TxS therapy that was a reversible thymidine auxotroph. To assess the impact of thymidine auxotrophy in the NTHi-host interplay during respiratory infection, thyA mutants were generated in both the clinical isolate NTHi375 and the reference strain RdKW20. Inactivation of the thyA gene increased TxS resistance, but also promoted morphological changes consistent with elongation and impaired bacterial division, which altered H. influenzae self-aggregation, phosphorylcholine level, C3b deposition, and airway epithelial infection patterns. Availability of external thymidine contributed to overcome such auxotrophy and TxS effect, potentially facilitated by the nucleoside transporter nupC. Although, thyA inactivation resulted in bacterial attenuation in a lung infection mouse model, it also rendered a lower clearance upon a TxS challenge in vivo. Thus, our results show that thymidine auxotrophy modulates both the NTHi host airway interplay and antibiotic resistance, which should be considered at the clinical setting for the consequences of TxS administration

Inactivation of the thymidylate synthase thyA in non-typeable Haemophilus influenzae modulates antibiotic resistance and has a strong impact on its interplay with the host airways.

Antibacterial treatment with cotrimoxazol (TxS), a combination of trimethoprim and sulfamethoxazole, generates resistance by, among others, acquisition of thymidine auxotrophy associated with mutations in the thymidylate synthase gene thyA, which can modify the biology of infection. The opportunistic pathogen non-typeable Haemophilus influenzae (NTHi) is frequently encountered in the lower airways of chronic obstructive pulmonary disease (COPD) patients, and associated with acute exacerbation of COPD symptoms. Increasing resistance of NTHi to TxS limits its suitability as initial antibacterial against COPD exacerbation, although its relationship with thymidine auxotrophy is unknown. In this study, the analysis of 2,542 NTHi isolates recovered at Bellvitge University Hospital (Spain) in the period 2010-2014 revealed 119 strains forming slow-growing colonies on the thymidine low concentration medium Mueller Hinton Fastidious, including one strain isolated from a COPD patient undergoing TxS therapy that was a reversible thymidine auxotroph. To assess the impact of thymidine auxotrophy in the NTHi-host interplay during respiratory infection, thyA mutants were generated in both the clinical isolate NTHi375 and the reference strain RdKW20. Inactivation of the thyA gene increased TxS resistance, but also promoted morphological changes consistent with elongation and impaired bacterial division, which altered H. influenzae self-aggregation, phosphorylcholine level, C3b deposition, and airway epithelial infection patterns. Availability of external thymidine contributed to overcome such auxotrophy and TxS effect, potentially facilitated by the nucleoside transporter nupC. Although, thyA inactivation resulted in bacterial attenuation in a lung infection mouse model, it also rendered a lower clearance upon a TxS challenge in vivo. Thus, our results show that thymidine auxotrophy modulates both the NTHi host airway interplay and antibiotic resistance, which should be considered at the clinical setting for the consequences of TxS administration