Mortality and cardiovascular risk in vasculitis ANCA. Importance of hypertension and renal function. Experience from southern Spain / Mortalidad y riesgo cardiovascular en vasculitis ANCA. Importancia de la hipertension y funcion renal. Experiencia del sur de España

Background: Cardiovascular disease (CVD) is one of the principal causes of death in antineutrophil cytoplasmic antibody-(ANCA)-associated vasculitis (AAV). Objectives: To evaluate the mortality and it's causes and CVD and its vascular risk factors (VRFs) in AAV patients in Andalusia. Methods: A multicenter cohort of 220 AAV patients followed-up from 1979 until June 2020 was studied in Andalussia, south of Spain. The information, including socio-demographic and clinical data was recorded retrospectively through chart review. Data was analysed using Chi2, ANOVA and Cox proportional hazards regresion as uni and multivariate test with a 95% confidence interval (CI). Results: During a mean ± standard deviation follow-up of 96.79 ± 75.83 months, 51 patients died and 30 presented at least one CVE. Independent prognostic factors of mortality were age (HR 1.083, p=0.001) and baseline creatinine (HR 4.41, p=0.01). Independent prognostic factors of CVE were age [hazard ratio (HR) 1.042, p=0.005] and the presence of hypertension (HTN) six months after diagnosis (HR 4.641, p=0.01). HTN, diabetes and renal failure, all of these important VRFs, are more prevalent in AAV patients than it is described in matched general population. Conclusions: Age and baseline renal function, but not CVEs, are predictors of mortality and age and early HTN are independent predictors for having a CVE. CVD screening in AAV patients is demanded.(AU)

Introducción: La enfermedad cardiovascular (ECV) es una de las principales causas de muerte en las vasculitis asociadas a anticuerpos anticitoplasma de neutrófilos (ANCA) (VAA). Objetivos: Evaluar la mortalidad y sus causas, entre ellas la ECV y sus factores de riesgo vascular (FRV) en pacientes con VAA en Andalucía. Métodos: Se estudió una cohorte multicéntrica de 220 pacientes con VAA seguidos desde 1979 hasta junio de 2020 en Andalucía. La información, incluidos los datos sociodemográficos y clínicos, se registró retrospectivamente a través de la revisión de historias clínicas. Los datos se analizaron mediante Chi2, ANOVA y regresión de riesgos proporcionales de Cox de forma uni y multivariante con un intervalo de confianza (IC) del 95%. Resultados: Durante un seguimiento medio y desviación estándar de 96,79 ± 75,83 meses, 51 pacientes fallecieron y 30 presentaron al menos un ECV. Los factores pronósticos independientes de mortalidad fueron la edad (HR 1,083, p=0,001) y la creatinina basal (HR 4,41, p=0,01). Los factores pronósticos independientes de ECV fueron la edad [hazard ratio (HR) 1,042, p=0,005] y la presencia de hipertensión arterial (HTA) seis meses después del diagnóstico (HR 4,641, p=0,01). La prevalencia de HTA, diabetes e insuficiencia renal fue elevada o muy elevada en comparación con la población general emparentada, todos FRCV determinantes para el pronóstico de estos pacientes. Conclusiones: La edad y la función renal basal son predictores de mortalidad y la edad y la HTA de aparición precoz son predictores independientes de tener ECV. Se recomienda el cribado de FRCV en pacientes con vasculitis ANCA.(AU)

Mortality and cardiovascular risk in vasculitis ANCA. Importance of hypertension and renal function. Experience from southern Spain.

BACKGROUND: Cardiovascular disease (CVD) is one of the principal causes of death in antineutrophil cytoplasmic antibody-(ANCA)-associated vasculitis (AAV). OBJECTIVES: To evaluate the mortality and it's causes and CVD and its vascular risk factors (VRFs) in AAV patients in Andalusia. METHODS: A multicenter cohort of 220 AAV patients followed-up from 1979 until June 2020 was studied in Andalussia, south of Spain. The information, including socio-demographic and clinical data was recorded retrospectively through chart review. Data was analysed using Chi2, ANOVA and Cox proportional hazards regresion as uni and multivariate test with a 95% confidence interval (CI). RESULTS: During a mean ± standard deviation follow-up of 96.79 ± 75.83 months, 51 patients died and 30 presented at least one CVE. Independent prognostic factors of mortality were age (HR 1.083, p=0.001) and baseline creatinine (HR 4.41, p=0.01). Independent prognostic factors of CVE were age [hazard ratio (HR) 1.042, p=0.005] and the presence of hypertension (HTN) six months after diagnosis (HR 4.641, p=0.01). HTN, diabetes and renal failure, all of these important VRFs, are more prevalent in AAV patients than it is described in matched general population. CONCLUSIONS: Age and baseline renal function, but not CVEs, are predictors of mortality and age and early HTN are independent predictors for having a CVE. CVD screening in AAV patients is demanded.

Long-term treatment with nebulized colistin in oncological patients with recurrent respiratory infections

Objectives: To assess the efficacy of long-term treatment with nebulized colistin in reducing the number of respiratory infections, emergency consultations and hospitalizations in oncological patients.MethodsA retrospective, observational, single-centre study including patients with solid or haematologic malignancies, or pulmonary GVHD after HSTC who received treatment with nebulized colistin for at least six-months to prevent recurrent respiratory infections (July 2010 to June 2017).ResultsTwelve patients were included (median age: 54.4, range: 23–85), 7 with solid malignancies and 5 with haematologic malignancies (2 with pulmonary GVHD). Pseudomonas aeruginosa was the most frequent microorganism in sputum cultures (11/12 patients), all strains were susceptible to colistin. There was a statistically significant reduction (p=0.01) in respiratory infections in the six-month period after starting colistin (median: 1, range: 0–4) compared to the six-month period before (median: 4, range: 1–8). There was also a reduction in emergency consultations (precolistin: median: 1.50, range: 0–3; postcolistin: median: 0, range: 0–3) and hospitalizations (precolistin: median: 1.50, range: 0–3; postcolistin: median: 0, range: 0–3) due to respiratory infections. No colistin-resistant strains were identified.ConclusionsLong-term treatment with nebulized colistin may be useful to reduce the number of exacerbations in oncological patients with recurrent respiratory infections. (AU)

Objetivos: Evaluar la eficacia de un tratamiento prolongado con colistina nebulizada para reducir el número de infecciones respiratorias, consultas en Urgencias y hospitalizaciones en pacientes oncológicos.MétodosEstudio retrospectivo, observacional y unicéntrico en pacientes con neoplasias sólidas o hematológicas o EICR pulmonar tras TPH tratados con colistina nebulizada al menos 6 meses para prevenir infecciones respiratorias recurrentes (julio del 2010-junio del 2017).ResultadosSe incluyó a 12 pacientes (edad mediana 54,4, rango: 23-85), 7 con cáncer sólido y 5 con neoplasias hematológicas (2 con EICR pulmonar). El microorganismo aislado más frecuentemente en esputos fue Pseudomonasaeruginosa (11/12 pacientes); todas las cepas fueron colistina-sensibles. Se evidenciaron una reducción estadísticamente significativa (p = 0,01) de las infecciones respiratorias en los 6 meses tras iniciar colistina (mediana: 1, rango: 0-4) comparado con los 6 meses previos (mediana: 4, rango: 1-8), y una reducción del número de visitas a Urgencias (precolistina: mediana: 1,50, rango: 0-3; postcolistina: mediana: 0, rango: 0-3) y hospitalizaciones (precolistina: mediana: 1,50, rango: 0-3; postcolistina: mediana: 0, rango: 0-3) por infección respiratoria. No se detectaron cepas resistentes a colistina.ConclusionesUn tratamiento prolongado con colistina nebulizada puede ser útil para reducir el número de exacerbaciones en pacientes oncológicos con infecciones respiratorias recurrentes. (AU)

An Immunochemical Approach to Detect the Quorum Sensing-Regulated Virulence Factor 2-Heptyl-4-Quinoline N-Oxide (HQNO) Produced by Pseudomonas aeruginosa Clinical Isolates.

Understanding quorum sensing (QS) and its role in the development of pathogenesis may provide new avenues for diagnosing, surveillance, and treatment of infectious diseases. For this purpose, the availability of reliable and efficient analytical diagnostic tools suitable to specifically detect and quantify these essential QS small molecules and QS regulated virulence factors is crucial. Here, we reported the development and evaluation of antibodies and an enzyme-linked immunosorbent assay (ELISA) for HQNO (2-heptyl-4-quinoline N-oxide), a QS product of the PqsR system, which has been found to act as a major virulence factor that interferes with the growth of other microorganisms. Despite the nonimmunogenic character of HQNO, the antibodies produced showed high avidity and the microplate-based ELISA developed could detect HQNO in the low nM range. Hence, a limit of detection (LOD) of 0.60 ± 0.13 nM had been reached in Müeller Hinton (MH) broth, which was below previously reported levels using sophisticated equipment based on liquid chromatography coupled to mass spectrometry. The HQNO profile of release of different Pseudomonas aeruginosa clinical isolates analyzed using this ELISA showed significant differences depending on whether the clinical isolates belonged to patients with acute or chronic infections. These data point to the possibility of using HQNO as a specific biomarker to diagnose P. aeruginosa infections and for patient surveillance. Considering the role of HQNO in inhibiting the growth of coinfecting bacteria, the present ELISA will allow the investigation of these complex bacterial interactions underlying infections. IMPORTANCE Bacteria use quorum sensing (QS) as a communication mechanism that releases small signaling molecules which allow synchronizing a series of activities involved in the pathogenesis, such as the biosynthesis of virulence factors or the regulation of growth of other bacterial species. HQNO is a metabolite of the Pseudomonas aeruginosa-specific QS signaling molecule PQS (Pseudomonas quinolone signal). In this work, the development of highly specific antibodies and an immunochemical diagnostic technology (ELISA) for the detection and quantification of HQNO was reported. The ELISA allowed profiling of the release of HQNO by clinical bacterial isolates, showing its potential value for diagnosing and surveillance of P. aeruginosa infections. Moreover, the antibodies and the ELISA reported here may contribute to the knowledge of other underlying conditions related to the pathology, such as the role of the interactions with other bacteria of a particular microbiota environment.

Long-term treatment with nebulized colistin in oncological patients with recurrent respiratory infections.

OBJECTIVES: To assess the efficacy of long-term treatment with nebulized colistin in reducing the number of respiratory infections, emergency consultations and hospitalizations in oncological patients. METHODS: A retrospective, observational, single-centre study including patients with solid or haematologic malignancies, or pulmonary GVHD after HSTC who received treatment with nebulized colistin for at least six-months to prevent recurrent respiratory infections (July 2010 to June 2017). RESULTS: Twelve patients were included (median age: 54.4, range: 23-85), 7 with solid malignancies and 5 with haematologic malignancies (2 with pulmonary GVHD). Pseudomonas aeruginosa was the most frequent microorganism in sputum cultures (11/12 patients), all strains were susceptible to colistin. There was a statistically significant reduction (p=0.01) in respiratory infections in the six-month period after starting colistin (median: 1, range: 0-4) compared to the six-month period before (median: 4, range: 1-8). There was also a reduction in emergency consultations (precolistin: median: 1.50, range: 0-3; postcolistin: median: 0, range: 0-3) and hospitalizations (precolistin: median: 1.50, range: 0-3; postcolistin: median: 0, range: 0-3) due to respiratory infections. No colistin-resistant strains were identified. CONCLUSIONS: Long-term treatment with nebulized colistin may be useful to reduce the number of exacerbations in oncological patients with recurrent respiratory infections.

Pertactin-Deficient Bordetella pertussis with Unusual Mechanism of Pertactin Disruption, Spain, 1986-2018.

Bordetella pertussis not expressing pertactin has increased in countries using acellular pertussis vaccines (ACV). The deficiency is mostly caused by pertactin gene disruption by IS481. To assess the effect of the transition from whole-cell vaccine to ACV on the emergence of B. pertussis not expressing pertactin in Spain, we studied 342 isolates collected during 1986-2018. We identified 93 pertactin-deficient isolates. All were detected after introduction of ACV and represented 38% of isolates collected during the ACV period; 58.1% belonged to a genetic cluster of isolates carrying the unusual prn::del(-292, 1340) mutation. Pertactin inactivation by IS481 insertion was identified in 23.7% of pertactin-deficient isolates, arising independently multiple times and in different phylogenetic branches. Our findings support the emergence and dissemination of a cluster of B. pertussis with an infrequent mechanism of pertactin disruption in Spain, probably resulting from introduction of ACV.

Empleo de la MAPA de 48 horas en atención especializada. Experiencia en nuestro centro / Use of 48-hour ABPM in specialist care. Experience in our centre

La monitorización ambulatoria de la presión arterial (MAPA) es una herramienta básica en el diagnóstico y tratamiento de la hipertensión arterial (HTA). La validez se basa en el número de mediciones realizadas y en su duración. Nuestro objetivo es estudiar en nuestra serie de MAPA de 48 h, si existe justificación para ampliar la duración de la MAPA a 48 h. Material y métodos: Análisis retrospectivo de una serie de casos formada por 81 pacientes con MAPA de 48 h durante un periodo de cinco años (2013 a 2018). Se analizan las diferencias entre el primer y segundo día. Resultados: Ochenta y un pacientes, 44 hombres, edad media 52 años (± 18). Mayor promedio de presión arterial (PA) en el primer día (132/77 mmHg vs. 130/76 mmHg, p ≤ 0,01) y también mayor proporción de pacientes con HTA el primer día (59 vs. 50%; p ≤ 0,05). Los pacientes con enfermedad renal crónica (ERC) (n = 33) presentaron PA sistólica (PAS) mayor en la segunda noche (p ≤ 0,05), un patrón circadiano de mayor riesgo en el segundo día (dipper 13,6 vs. 86,4%, nondipper 60,7 vs. 41,7% y riser 30,3 vs. 18,8%; p ≤ 0,05), más diabetes (39%, p ≤ 0,01) y más hipertrofia del ventrículo izquierdo (HVI) (74%, p ≤ 0,05). Conclusiones: La MAPA de 48 h podría determinar mejor los valores de PA y patrón circadiano que la de 24 h, en especial a los pacientes con ERC y diabetes, ambas patologías de elevado riesgo cardiovascular.(AU)

Ambulatory blood pressure monitoring (ABPM) is a basic tool in the diagnosis and treatment of hypertension (HT). Validity is based on the number of readings taken and their duration. Our aim was to study in our 48-hour ABPM series whether extending the duration of ABPM to 48 hours is justified. Material and methods: Retrospective analysis of a case series comprising 81 patients with 48-hour ABPM over a 5-year period (2013 to 2018). We analysed the differences between the first and second day. Results: Eighty-one patients, 44 men, mean age of 52 years (± 18). The mean blood pressure (BP) was higher on the first day (132/77 mmHg vs. 130/76 mmHg, p ≤ .01) and there was also a greater proportion of patients with HT on the first day (59 vs. 50%; p ≤ .05). The patients with chronic kidney disease (CKD) (n = 33) had higher systolic BP (SBP) on the second night (p ≤ .05), a circadian rhythm of higher risk on the second day (dipper 13.6 vs. 86.4%, non-dipper 60.7 vs. 41.7%, and riser 30.3 vs. 18.8%; p ≤ .05), more diabetes (39%, p ≤ .01) and more left ventricular hypertrophy (LVH) (74%, p ≤ .05). Conclusions: 48-hour ABPM could determine BP readings and circadian rhythm better than 24-hour ABPM, especially in patients with CKD and diabetes, both diseases carrying high cardiovascular risk.(AU)

[Use of 48-hour ABPM in specialist care. Experience in our centre]. / Empleo de la MAPA de 48 horas en atención especializada. Experiencia en nuestro centro.

Ambulatory blood pressure monitoring (ABPM) is a basic tool in the diagnosis and treatment of hypertension (HT). Validity is based on the number of readings taken and their duration. Our aim was to study in our 48-hour ABPM series whether extending the duration of ABPM to 48 hours is justified. MATERIAL AND METHODS: Retrospective analysis of a case series comprising 81 patients with 48-hour ABPM over a 5-year period (2013 to 2018). We analysed the differences between the first and second day. RESULTS: Eighty-one patients, 44 men, mean age of 52 years (± 18). The mean blood pressure (BP) was higher on the first day (132/77 mmHg vs. 130/76 mmHg, p ≤ .01) and there was also a greater proportion of patients with HT on the first day (59 vs. 50%; p ≤ .05). The patients with chronic kidney disease (CKD) (n = 33) had higher systolic BP (SBP) on the second night (p ≤ .05), a circadian rhythm of higher risk on the second day (dipper 13.6 vs. 86.4%, non-dipper 60.7 vs. 41.7%, and riser 30.3 vs. 18.8%; p ≤ .05), more diabetes (39%, p ≤ .01) and more left ventricular hypertrophy (LVH) (74%, p ≤ .05). CONCLUSIONS: 48-hour ABPM could determine BP readings and circadian rhythm better than 24-hour ABPM, especially in patients with CKD and diabetes, both diseases carrying high cardiovascular risk.

A new protocol reduces median door-to-needle time to the benchmark of 30 minutes in acute stroke treatment.

INTRODUCTION: Recent analyses emphasise that The Benchmark Stroke Door-to-Needle Time (DNT) should be 30min. This study aimed to determine if a new in-hospital IVT protocol is effective in reducing door-to-needle time and correcting previously identified factors associated with delays. MATERIAL AND METHODS: In 2014, we gradually introduced a series of measures aimed to reduce door-to-needle time for patients receiving IVT, and compared it before (2009-2012) and after (2014-2017) the new protocol was introduced. RESULTS: The sample included 239 patients before and 222 after the introduction of the protocol. Median overall door-to-needle time was 27min after the protocol was fully implemented (a 48% reduction on previous door-to-needle time [52min], P<.001)]. Median door-to-needle time was lower when pre-hospital code stroke was activated (22min). We observed a 26-min reduction in the median time from onset to treatment (P<.001). After the protocol was implemented, the "3-hour-effect" did not affect door-to-needle time (P=.98). Computed tomography angiography studies performed before IVT were associated with increased door-to-needle time (P<.001); however, the test was performed after IVT was started in most cases. CONCLUSIONS: Hospital reorganisation and multidisciplinary collaboration brought median door-to-needle time below 30min and corrected previously identified delay factors. Furthermore, overall time from onset to treatment was also reduced and more stroke patients were treated within 90min of symptom onset.

Un nuevo protocolo intrahospitalario reduce el tiempo puerta-aguja en el ictus agudo tratado con trombolisis intravenosa a menos de 30 minutos / A new protocol reduces median door-to-needle time to the benchmark of 30 minutes in acute stroke treatment

Introducción: El objetivo del tiempo puerta-aguja en el ictus isquémico agudo tratado con trombólisis intravenosa (TIV) tiende a situarse actualmente en los 30 min. Determinamos si un nuevo protocolo de actuación intrahospitalario es eficaz para reducir el intervalo puerta-aguja y corregir los factores de demora previamente identificados.Material y métodosEn 2014 se implantaron gradualmente unas medidas diseñadas para acortar los tiempos de actuación intrahospitalarios en los pacientes tratados con TIV. Se compararon los tiempos de actuación antes (2009-2012) y después (febrero 2014-abril 2017) de la introducción del nuevo protocolo.ResultadosSe incluyeron 239 pacientes antes y 222 después. Cuando todas las medidas fueron introducidas, la mediana global de tiempo puerta-aguja fue de 27 min (previa 52 min, 48% menos, p < 0,001) y de 22 min cuando se activó el código ictus extrahospitalario. El tiempo global al tratamiento (inicio-aguja) se redujo en 26 min de mediana (p < 0,001). En el período postintervención ya no se objetivó el «efecto de fin de ventana» (p = 0,98). Aunque la angio-TC antes de la TIV continuó retrasando los tiempos de actuación (p < 0,001), tras el nuevo protocolo, esta prueba se realizó después del inicio del tratamiento en la mayoría de los casos.ConclusionesLa reorganización intrahospitalaria y la colaboración multidisciplinar han situado la mediana de tiempo puerta-aguja por debajo de los 30 min y han corregido los factores de demora identificados previamente. Además, se ha reducido el tiempo global al tratamiento y una mayor proporción de pacientes son tratados en los primeros 90 min desde el inicio de los síntomas. (AU)

Introduction: Recent analyses emphasize that The Benchmark Stroke Door-to-Needle Time (DNT) should be 30 min. This study aimed to determine if a new in-hospital IVT protocol is effective in reducing door-to-needle time and correcting previously identified factors associated with delays.Material and methodsIn 2014, we gradually introduced a series of measures aimed to reduce door-to-needle time for patients receiving IVT, and compared it before (2009-2012) and after (2014-2017) the new protocol was introduced.ResultsThe sample included 239 patients before and 222 after the introduction of the protocol. Median overall door-to-needle time was 27 min after the protocol was fully implemented (a 48% reduction on previous door-to-needle time [52 minutes], P<.001)]. Median door-to-needle time was lower when pre-hospital code stroke was activated (22 min). We observed a 26-min reduction in the median time from onset to treatment (P<.001). After the protocol was implemented, the «3-hour-effect» did not affect door-to-needle time time (P=.98). Computed tomography angiography studies performed before IVT were associated with increased door-to-needle time (P<.001); however, the test was performed after IVT was started in most cases.ConclusionsHospital reorganisation and multidisciplinary collaboration brought median door-to-needle time below 30 min and corrected previously identified delay factors. Furthermore, overall time from onset to treatment was also reduced and more stroke patients were treated within 90 min of symptom onset. (AU)

An Immunochemical Approach to Quantify and Assess the Potential Value of the Pseudomonas Quinolone Signal as a Biomarker of Infection.

Quorum sensing (QS) is a bacterial cell density-based communication system using low molecular weight signals called autoinducers (AIs). Identification and quantification of these molecules could provide valuable information related to the stage of colonization or infection as well as the stage of the disease. With this scenario, we report here for the first time the development of antibodies against the PQS (pseudomonas quinolone signal), the main signaling molecule from the pqs QS system of Pseudomonas aeruginosa, and the development of a microplate-based enzyme-linked immunosorbent assay (ELISA) able of quantifying this molecule in complex biological media in the low nanometer range (LOD, 0.36 ± 0.14 nM in culture broth media). Moreover, the PQS ELISA here reported has been found to be robust and reliable, providing accurate results in culture media. The technique allowed us to follow up the PQS profile of the release of bacterial clinical isolates obtained from patients of different disease status. A clear correlation was found between the PQS immunoreactivity equivalents and the chronic or acute infection conditions, which supports the reported differences on virulence and behavior of these bacterial strains due to their adaptation capability to the host environment. The results obtained point to the potential of the PQS as a biomarker of infection and to the value of the antibodies and the technology developed for improving diagnosis and management of P. aeruginosa infections based on the precise identification of the pathogen, appropriate stratification of the patients according to their disease status, and knowledge of the disease progression.

A new pleosporalean fungus isolated from superficial to deep human clinical specimens.

Human infections by pleosporalean fungi (class Dothideomycetes, phylum Ascomycota) are rarely reported. Because their identification is challenging using morphological characterization, several phylogenetic markers must be sequenced for an accurate identification and taxonomical placement of the isolates. Three isolates of clinical origin were phenotypically characterized, but due to the absence of relevant morphological traits, D1-D2 domains of the 28S nrRNA gene (LSU), the internal transcribed spacer region (ITS) of the nrRNA, and fragments of the RNA polymerase II subunit 2 (rpb2) and translation elongation factor 1-alpha (tef1) genes were sequenced to allow a phylogenetic analysis that would solve their phylogenetic placement. That analysis revealed that these isolates did not match any previously known pleosporalean genera, and they are proposed here as the new fungal genus, Gambiomyces. Unfortunately, the isolates remained sterile, which, consequently, made the morphological description of the reproductive structures impossible. Future studies should try to understand the behaviour of this fungus in nature as well as its characteristics as an opportunistic fungal pathogen. Molecular identification is becoming an essential tool for proper identification of Dothideomycetes of clinical origin. LAY ABSTRACT: We describe a new pleosporalen pathogenic fungus, Gambiomyces profunda, found in superficial to deep samples from a human patient. Because all strains remained sterile, the fungus was finally identified following a phylogenetic analysis by using four different molecular markers.

Consensus Multilocus Sequence Typing Scheme for Pneumocystis jirovecii.

Pneumocystis jirovecii is an opportunistic human pathogenic fungus causing severe pneumonia mainly in immunocompromised hosts. Multilocus sequence typing (MLST) remains the gold standard for genotyping of this unculturable fungus. However, the lack of a consensus scheme impedes a global comparison, large scale population studies and the development of a global MLST database. To overcome this problem this study compared all genetic regions (19 loci) currently used in 31 different published Pneumocystis MLST schemes. The most diverse/commonly used eight loci, ß-TUB, CYB, DHPS, ITS1, ITS1/2, mt26S and SOD, were further assess for their ability to be successfully amplified and sequenced, and for their discriminatory power. The most successful loci were tested to identify genetically related and unrelated cases. A new consensus MLST scheme consisting of four genetically independent loci: ß-TUB, CYB, mt26S and SOD, is herein proposed for standardised P. jirovecii typing, successfully amplifying low and high fungal burden specimens, showing adequate discriminatory power, and correctly identifying suspected related and unrelated isolates. The new consensus MLST scheme, if accepted, will for the first time provide a powerful tool to investigate outbreak settings and undertake global epidemiological studies shedding light on the spread of this important human fungal pathogen.

High-Throughput Immunochemical Method to Assess the 2-Heptyl-4-quinolone Quorum Sensing Molecule as a Potential Biomarker of Pseudomonas aeruginosa Infections.

Bacterial quorum sensing (QS) is being contemplated as a promising target for developing innovative diagnostic and therapeutic strategies. Here we report for the first time the development of antibodies against 2-heptyl-4-quinolone (HHQ), a signaling molecule from the pqs QS system of Pseudomonas aeruginosa, involved in the production of important virulent factors and biofilm formation. The antibodies produced were used to develop an immunochemical diagnostic approach to assess the potential of this molecule as a biomarker of P. aeruginosa infection. The ELISA developed is able to reach a detectability in the low nM range (IC50 = 4.59 ± 0.29 nM and LOD = 0.34 ± 0.13 nM), even in complex biological samples such as Müeller Hinton (MH) culture media. The ELISA developed is robust and reproducible and has been found to be specific to HHQ, with little interference from other related alkylquinolones from the pqs QS system. The ELISA has been used to analyze the HHQ production kinetics of P. aeruginosa clinical isolates grown in MH media, pointing to its potential as a biomarker of infection and at the possibility to use the technology developed to obtain additional information about the disease stage.

Isolated cerebral mucormycosis associated with intravenous drug use.

We present an uncommon case of isolated basal ganglia mucormycosis in a patient without any known cause of immunosuppression, but with a history of drug injection. The patient presented a good clinical and radiological response to antifungal treatment without aggressive surgical debridement (liposomal amphotericin B combined with isavuconazole for 4 weeks followed by isavuconazole as maintenance therapy for 10 months).

Detection of Streptococcus pneumoniae antigen in pleural fluid: usefulness of an immunofluorescence-based lateral flow assay for the diagnosis of pneumococcal pneumonia.

The performance of an immunofluorescence-based Streptococcus pneumoniae antigen detection test in pleural fluid (IF-PF) was evaluated. For proven and possible pneumococcal pneumonias global sensitivity and specificity were 92.6 (95 CI 76.6-97.9) and 80 (95 CI 62.7-90.5), respectively, with no significant differences between children and adults. Global diagnostic accuracy of IF-PF was 86% (74.2-93.7), and a substantial k index of concordance with culture/RT-PCR of 0.716 (0.535-0.896). IF-PF might be useful as a rapid complementary test for the etiologic diagnosis of pneumococcal pneumonia.

Timely Diagnosis of Histoplasmosis in Non-endemic Countries: A Laboratory Challenge.

Human histoplasmosis is a fungal infection caused by the inhalation of microconidia of the thermally dimorphic fungi Histoplasma capsulatum. Autochthonous cases of histoplasmosis have been diagnosed in almost every country, but it is considered an endemic infection in specific areas of the world. Many of them are popular travel destinations or the source of migratory movements. Thus, the vast majority of the registered cases in non-endemic countries are imported. They correspond to people having been exposed to the fungus in endemic locations as immigrants, expatriates, transient workers or tourists, with reported cases also associated to organ donation. Misdiagnosis and delays in initiation of treatment are not uncommon in cases of imported histoplasmosis. They are associated to high fatality-rates specially in patients with compromised cellular immunity in which progressive disseminated forms develop. The diagnosis of this infection in non-endemic countries is hampered by the lack of clinical suspicion and a dearth of available diagnostic tools adequate to offer rapid and accurate results. Non-culture-based assays such as nucleic-acid amplification tests present as a suitable alternative in this situation, offering improved sensitivity and specificity, shortened turnaround time, and increased biosafety by avoiding culture manipulation. In non-endemic regions, molecular techniques are being used mainly in laboratories from countries that have registered an increase in the incidence of imported cases. However, the number of published techniques is limited and lack consensus. Efforts are currently under way to standardize nucleic acid amplification-based techniques for its implementation in areas registering a rising number of imported cases.

Taking a look on fungi in cystic fibrosis: more questions than answers / Una mirada a los hongos en la fibrosis quística: más preguntas que respuestas

Cystic fibrosis (CF) is one of the most frequent recessive inherited diseases in western countries. Advances in medical care have led to a substantial increase in the life expectancy of CF patients. Survival beyond adolescence has permitted to see fungi not only as late colonizers, but also as potential pathogens responsible of allergic reactions and chronic infections related to lung function deterioration. The role of fungi, nevertheless, has been overlooked until recently. As a result, a number of questions on their epidemiology, clinical significance, or diagnosis, among others, remain unanswered. Besides more in depth studies about the extent of the deleterious effect of fungi on the CF host, new technologies may provide the key to understand its pathogenic role, its interaction with other microbial components of the respiratory microbiota, and should pave the way to define subsets of patients at risk who would benefit from specific therapy. This review is intended to provide a quick overview on what we know about the presence of fungi in the CF airway and its repercussion in the host, and to point out some of the many knowledge gaps needed to understand and advance in the management of fungi in the airway of CF subjects

La fibrosis quística (FQ) es una de las enfermedades genéticas recesivas más frecuentes en los países occidentales. Los avances médicos han supuesto un notable incremento de la esperanza de vida de estos pacientes. La supervivencia más allá de la adolescencia ha revelado la presencia de los hongos no solo como colonizadores tardíos sino como patógenos potenciales, responsables de reacciones alérgicas e infecciones crónicas relacionadas con el deterioro de la función pulmonar. El papel de los hongos en los pacientes con FQ, no obstante, ha sido pasado por alto hasta fechas recientes. Como consecuencia, quedan por responder numerosas cuestiones sobre su epidemiología, significado clínico o diagnóstico, entre otras. Son precisos estudios en profundidad sobre el verdadero alcance del efecto deletéreo de los hongos en el paciente con FQ. Las nuevas tecnologías pueden resultar claves para entender su papel patogénico, su interacción con otros componentes de la microbiota respiratoria, y para la identificación de los subgrupos de pacientes de más riesgo que se beneficiarían particularmente de las terapias dirigidas. Esta revisión pretende proporcionar una rápida mirada a lo que sabemos actualmente sobre la presencia de los hongos en las vías respiratorias de los pacientes FQ y su repercusión en el huésped, así como señalar las múltiples lagunas de conocimiento que es necesario rellenar para entender y avanzar en el manejo de la enfermedad fúngica respiratoria de los pacientes con FQ

Tumor neuroendocrino apendicular pediátrico: análisis de una serie / Pediatric neuroendocrine appendicular tumor: a case-series analysis

INTRODUCCIÓN: El tumor neuroendocrino es la neoplasia apendicular más frecuente en la edad pediátrica. Existen guías sobre su manejo en adultos pero no se han elaborado guías para pacientes pediátricos, lo que conlleva un manejo muy heterogéneo en este grupo de edad. Se presenta una serie de casos con el objetivo de mejorar el conocimiento de estos tumores. MATERIAL Y MÉTODOS: Se realizó un estudio retrospectivo de los casos de tumor neuroendocrino apendicular hallados en piezas de apendicectomía entre enero de 2004 y diciembre de 2017 en dos hospitales terciarios, en menores de 18 años. RESULTADOS: se incluyeron 6 pacientes, 4 varones y 2 mujeres. En todos el síntoma principal fue el dolor abdominal, ninguno presentó síndrome carcinoide. En uno de ellos el tumor se localizó en base y tuvo dudosa afectación de márgenes, por lo que se realizó una segunda cirugía. En el resto, la apendicectomía fue curativa. Sólo en 3 se hizo seguimiento. CONCLUSIONES: según la mayoría de autores no es necesaria una segunda cirugía tras el diagnóstico, ni tampoco el seguimiento pues se han encontrado supervivencias similares cuando se comparan grupos. Esta afirmación parece aún más clara en tumores <1 cm y sin factores de riesgo asociados, sobre todo la invasión del mesoapéndice. En cualquier caso, hacen falta estudios para la elaboración de guías que permitan homogeneizar su manejo en la edad pediátrica

INTRODUCTION: The neuroendocrine tumor is the most frequent appendix neoplasia in the pediatric age. Although multiple guides about the management of this tumor in adults exist, no guides with focus on children have been created. That makes the management of the tumor for this population very heterogeneous. Several cases are presented below with the objective to improve the knowledge on this area. MATERIAL AND METHODS: a retrospective study of the appendix neuroendocrine tumor found in appendectomy pieces on patients under eighteen was carried out in two tertiary hospitals - Nuestra Señora del Prado in Talavera de la Reina and Virgen de la Salud in Toledo- between January 2004 and December 2017. RESULTS: six patients were under study, 4 men and 2 women. In all of them, the main symptom was abdominal pain, any of them showed carcinoid syndrome. One had a tumor located in the base. In this case a second surgery was necessary due to the suspicion of affected margins. Regarding the rest of the patients, the appendectomy was curative. Only in three of them a follow up was done. CONCLUSIONS: According to the majority of the authors, neither a second surgery nor a follow up is necessary after the diagnosis, having similar results when comparing groups. This statement becomes more significant in tumors <1 cm and without associated risk factors, specially the deep masoappendiceal infiltration. In any case, it is clear that further studies must be conducted in order to elaborate guides that allow making the management of the tumor more homogeneous

Taking a look on fungi in cystic fibrosis: More questions than answers.

Cystic fibrosis (CF) is one of the most frequent recessive inherited diseases in western countries. Advances in medical care have led to a substantial increase in the life expectancy of CF patients. Survival beyond adolescence has permitted to see fungi not only as late colonizers, but also as potential pathogens responsible of allergic reactions and chronic infections related to lung function deterioration. The role of fungi, nevertheless, has been overlooked until recently. As a result, a number of questions on their epidemiology, clinical significance, or diagnosis, among others, remain unanswered. Besides more in depth studies about the extent of the deleterious effect of fungi on the CF host, new technologies may provide the key to understand its pathogenic role, its interaction with other microbial components of the respiratory microbiota, and should pave the way to define subsets of patients at risk who would benefit from specific therapy. This review is intended to provide a quick overview on what we know about the presence of fungi in the CF airway and its repercussion in the host, and to point out some of the many knowledge gaps needed to understand and advance in the management of fungi in the airway of CF subjects.