New insights from short and long reads sequencing to explore cytochrome b variants in Plasmopara viticola populations collected from vineyards and related to resistance to complex III inhibitors.

Downy mildew is caused by Plasmopara viticola, an obligate oomycete plant pathogen, a devasting disease of grapevine. To protect plants from the disease, complex III inhibitors are among the fungicides widely used. They specifically target the mitochondrial cytochrome b (cytb) of the pathogen to block cellular respiration mechanisms. In the French vineyard, P. viticola has developed resistance against a first group of these fungicides, the Quinone outside Inhibitors (QoI), with a single amino acid substitution G143A in its cytb mitochondrial sequence. The use of QoI was limited and another type of fungicide, the Quinone inside Inhibitors, targeting the same gene and highly effective against oomycetes, was used instead. Recently however, less sensitive P. viticola populations were detected after treatments with some inhibitors, in particular ametoctradin and cyazofamid. By isolating single-sporangia P. viticola strains resistant to these fungicides, we characterized new variants in the cytb sequences associated with cyazofamid resistance: a point mutation (L201S) and more strikingly, two insertions (E203-DE-V204, E203-VE-V204). In parallel with the classical tools, pyrosequencing and qPCR, we then benchmarked short and long-reads NGS technologies (Ion Torrent, Illumina, Oxford Nanopore Technologies) to sequence the complete cytb with a view to detecting and assessing the proportion of resistant variants of P. viticola at the scale of a field population. Eighteen populations collected from French vineyard fields in 2020 were analysed: 12 showed a variable proportion of G143A, 11 of E203-DE-V204 and 7 populations of the S34L variant that confers resistance to ametoctradin. Interestingly, the long reads were able to identify variants, including SNPs, with confidence and to detect a small proportion of P. viticola with multiple variants along the same cytb sequence. Overall, NGS appears to be a promising method for assessing fungicide resistance of pathogens linked to cytb modifications at the field population level. This approach could rapidly become a robust decision support tool for resistance management in the future.

Tasa de egreso hospitalario por episodio depresivo recurrente en los años 2018-2021 en Chile / Hospital discharge rate due to recurrent depressive episode in the years 2018-2021 in Chile

Introducción: La depresión es un trastorno del ánimo frecuente, cuya recurrencia altera su manejo y pronóstico. El objetivo del trabajo es describir la tasa de egreso hospitalario (TEH) por episodio depresivo recurrente (EDR) entre 2018-2021 en Chile, según sexo, edad y gravedad. Materiales y métodos: Estudio observacional y transversal que considera los egresos hospitalarios por EDR en el período 2018-2021 en Chile (N=1551). Se utilizaron las bases de datos del Departamento de Estadísticas e Información de Salud y el Instituto Nacional de Estadística. Las variables estudiadas fueron sexo, grupo etario y gravedad. No requirió aprobación de comité de ética. Resultados: Se observó la menor TEH en 2020 con 1,91 egresos por cada 100.000 habitantes. Las mujeres sobresalieron durante todo el período con una TEH de 3,68 egresos por cada 100.000 habitantes. Destacó el grupo de 15 a 19 años con la mayor TEH en ambos sexos con 5,3 egresos por cada 100.000 habitantes. Respecto a gravedad, las hospitalizaciones se concentraron en pacientes de diagnóstico no especificado. Discusión: La pandemia de COVID 19 podría explicar la disminución de la TEH en 2020, al reducirse el diagnóstico y hospitalización por EDR; presumiblemente debido al fenómeno de reconversión de camas. La TEH es mayor en el sexo femenino, lo cual es concordante con la literatura. El predominio del grupo de 15 a 19 años discrepa de la evidencia, la cual indica que suele concentrarse en individuos de 25 a 64 años. Conclusión: Las TEH por EDR se concentraron en mujeres jóvenes. Es relevante conocer la epidemiología local para focalizar los recursos en la detección oportuna de factores de riesgo, para evitar episodios graves y disminuir su recurrencia.

Introduction: Depression is a common mood disorder, whose recurrence alters its management and prognosis. The aim of the paper is to describe the hospital discharge rate (HED) for recurrent depressive episode (RDE) between 2018-2021 in Chile, according to sex, age and severity. Materials and methods: Observational and cross-sectional study considering hospital discharges due to DRE in the period 2018-2021 in Chile (N=1551). The databases of the Department of Health Statistics and Information and the National Institute of Statistics were used. The variables studied were sex, age group and severity. Ethics committee approval was not required. Results: The lowest HTE was observed in 2020 with 1.91 discharges per 100,000 inhabitants. Females stood out during the entire period with an HTE of 3.68 discharges per 100,000 population. The 15-19 years age group stood out with the highest HTE in both sexes with 5.3 admissions per 100,000 inhabitants. In terms of severity, hospitalizations were concentrated in patients with unspecified diagnosis. Discussion: The COVID 19 pandemic could explain the decrease in HTE in 2020, with a reduction in diagnosis and hospitalization for RDE, presumably due to the bed conversion phenomenon. HTE is higher in the female sex, which is consistent with the literature. The predominance of the 15 to 19 years age group disagrees with the evidence, which indicates that it tends to be concentrated in individuals aged 25 to 64 years. Conclusion: HTE due to DRE was concentrated in young women. It is important to know the local epidemiology in order to focus resources on the timely detection of risk factors to avoid serious episodes and reduce their recurrence.

Percepciones que tienen trabajadores chilenos sobre el impacto del teletrabajo en el entorno de covid-19

RESUMEN La pandemia ha generado diversos cambios que han llevado a las empresas a buscar estrategias para continuar sus procesos productivos y proteger la salud de los trabajadores; entre estas, la más destacada es el Teletrabajo. Objetivo: Analizar las percepciones que tienen los trabajadores chilenos sobre el impacto del teletrabajo en términos de Estrés laboral, Balance Vida-Trabajo y Satisfacción, en ambiente de contingencia producto del COVID-19. Material y Método: Investigación de tipo cuantitativa, tipo relacional y de corte transversal que utilizó un muestreo por conveniencia y se obtuvieron datos de 225 trabajadores chilenos que estaban realizando teletrabajo producto del COVID-19 y respondieron la encuesta on-line. Se utilizó un instrumento con 26 reactivos, que se aplicó a trabajadores mayores de edad que estuvieran realizando actividades desde casa, entre los meses de septiembre a octubre de 2020, logrando obtener 181 respuestas útiles. Resultados: Los trabajadores que se encuentran en modalidad telemática corresponden mayormente a mujeres, solteras, entre 24 a 40 años, del ámbito educativo, del sector público y tienen experiencia laboral mayor a diez años; existe una relación positiva entre la satisfacción laboral y el teletrabajo (r= 0,381), mientras que para las variables Estrés laboral y Balance vida-trabajo sucede lo contrario, es decir, una relación negativa (r= -0,408). Conclusión: Las actividades telemáticas producen a los trabajadores sintomatología de estrés, como tensión y/o ansiedad, pero a pesar de esto, tienen una actitud positiva ante este cambio en la modalidad laboral, tratando de vivir el día a día y buscando cómo apoyar a otros.

ABSTRACT The pandemic has caused several changes that have led companies to look for strategies to continue their production processes and protect the health of workers. Among these strategies, the most important is Teleworking. Objective: To analyze the perceptions that Chilean workers have on the impact of teleworking in terms of work stress, work-life balance and satisfaction, in a contingency environment as a result of COVID-19. Material and Method: Quantitative, relational and cross-sectional research using convenience sampling and data from 225 Chilean workers who were teleworking as a result of COVID-19 and responded to the on-line survey. An instrument with 26 items was applied to workers of legal age who were carrying out activities from home, between the months of September to October 2020, collecting 181 useful responses. Results: People working from home correspond mostly to single women, between 24 to 40 years old, who work in the field of education or the public sector and have work experience of more than ten years; there is a positive relationship between job satisfaction and teleworking (r= 0.381), while for the variables work stress and work-life balance a negative relationship was observed (r= -0.408). Conclusion: Teleworking produces stress symptoms among workers, such as tension and/or anxiety. Nevertheless, workers have a positive attitude towards this modality, trying to adapt and seeking support from other people.

RESUMO A pandemia gerou várias mudanças que levaram as empresas a procurar estratégias para continuar os seus processos de produção e proteger a saúde dos trabalhadores; entre estas, a mais relevante é o Teletrabalho. Objetivo: Analisar as percepções dos trabalhadores chilenos sobre o impacto do teletrabalho em termos de Estresse no trabalho, Equilíbrio Vida-Trabalho e Satisfação, em um ambiente de contingência resultante da COVID-19. Material e Método: Pesquisa quantitativa, do tipo relacional e corte transversal utilizando amostragem de conveniência e obtendo dados de 225 trabalhadores chilenos que estavam realizando teletrabalho como resultado da COVID-19 e responderam ao questionário online. Foi utilizado um instrumento com 26 reativos, que foi aplicado a trabalhadores adultos que realizavam atividades desde casa, entre setembro e outubro de 2020, obtendo 181 respostas úteis. Resultados: Os trabalhadores que estão no modo telemático, correspondem maioritariamente a mulheres, solteiras, entre 24 e 40 anos, do campo educativo, do sector público e têm mais de dez anos de experiência de trabalho; existe uma relação positiva entre a satisfação profissional e o teletrabalho (r= 0,381), enquanto para as variáveis Estresse no trabalho e Equilíbrio Vida-Trabalho acontece o contrário, ou seja, uma relação negativa (r= -0,408). Conclusão: As atividades telemáticas produzem sintomas de estresse nos trabalhadores, tais como tensão e/ou ansiedade, mas apesar disso, têm uma atitude positiva em relação a esta mudança na modalidade de trabalho, tentando viver dia após dia e procurando formas de apoiar os outros.

Loss of T-Cell Multifunctionality and TCR-Vß Repertoire Against Epstein-Barr Virus Is Associated With Worse Prognosis and Clinical Parameters in HIV+ Patients.

Epstein-Barr virus (EBV) is an oncogenic virus associated with the development of aggressive and poor-prognosis B-cell lymphomas in patients infected with human immunodeficiency virus (HIV+ patients). The most important risk factors for these malignancies include immune dysfunction, chronic immune activation, and loss of T-cell receptor (TCR) repertoire. The combination of all these factors can favor the reactivation of EBV, malignant cell transformation, and clinical progression toward B-cell lymphomas. The overarching aim of this study was to evaluate the frequency, phenotype, functionality, and distribution of TCR clonotypes for EBV-specific T-cell subpopulations in HIV+ patients at different clinical stages and for HIV+ patients with B-cell lymphoma, as well as to establish their association with clinical variables of prognostic value. Factors were studied in 56 HIV+ patients at different clinical stages and in six HIV+ subjects with diagnosed B-cell lymphoma. We found a significant decrease in all subpopulations of EBV-specific CD4+ T cells from HIV+ patients at stage 3 and with B-cell lymphoma. EBV-specific effector CD8+ T cells, particularly effector memory cells, were also reduced in HIV+ patients with B-cell lymphoma. Interestingly, these cells were unable to produce IFN-γ and lacked multifunctionality in HIV+ patients. The TCR-Vß repertoire, which is key for protection against EBV in healthy individuals, was less diverse in HIV+ patients due to a lower frequency of TCR-Vß2+, Vß4+, Vß7.1+, Vß9+, Vß13.6+, Vß14+, Vß17+, Vß22+ CD4+, Vß14+, and Vß17+ CD8+ T cells. HIV+ patients with positive plasma EBV loads (EBV+HIV+) had a noteworthy decrease in the levels of both TNF-α+ and multifunctional TNF-α+/IL-2+ and TNF-α+/IFN-γ+ CD8+ T cells. Altogether, our findings demonstrate that HIV+ patients have significant alterations in the immune response to EBV (poor-quality immunity) that can favor viral reactivation, escalating the risk for developing EBV-associated B-cell lymphomas.

Emergence of boscalid-resistant strains of Erysiphe necator in French vineyards.

The grapevine powdery mildew Erysiphe necator (E. necator) is an obligate pathogen. Powdery mildew-diseased vines show an important reduction in plant size, winter hardiness and grape yield. Even a low-level infection with powdery mildew was shown to taint wine and ultimately reduce wine quality. For many years, succinate dehydrogenase inhibitor (SDHI) fungicides, mainly the new generation active ingredients (AIs) boscalid, penthiopyrad and fluopyram, have been widely used to control powdery mildew in grapevines. The repeated use of fungicides (mainly boscalid) has resulted in the emergence of resistant microorganisms such as Botrytis cinerea (B. cinerea). However, boscalid resistance was never observed in E. necator. In this study, a large-scale survey of French grapevine field populations of E. necator revealed many field populations with low sensitivity to boscalid. Single spore strains originating from collected resistant populations showed Half maximal effective concentration (EC50) values greater than 100 mg L-1, and strains originating from boscalid sensitive populations showed EC50 values lower than 1 mg L-1. The complete nucleotide sequences of the EnSdhB succinate dehydrogenase of sensitive and resistant single spore strains revealed that H242R and H242Y substitutions in the EnSdhB succinate dehydrogenase subunit conferred E. necator resistance to boscalid. No cross-resistance of E. necator strains bearing H242R and H242Y substitutions in EnSdhB succinate dehydrogenase to fluxapyroxad and fluopyram was noticed. Therefore, our results highlight the emergence of resistance to boscalid activity in French vineyards and warrant the need of the implementation of risk assessment strategies to maintain effective grapevine protection against powdery mildew.

First report of sporadic cases of Candida auris in Colombia.

BACKGROUND: Candida auris is a recently reported Candida species that is phenotypically similar to Candida haemulonii and related to hospital outbreaks. This organism can be misidentified as Candida haemulonii, Candida famata, Candida catenulata, or Rhodotorula glutinis by phenotypic approaches. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and DNA sequence analysis using internal transcribed spacer rDNA bar-coding provide an accurate identification. CASE REPORTS: Three cases of C. auris infection in patients with risk factors for fungal infection (one admitted to the intensive care unit, one with lymphoma, and one with HIV; all three with previous antibiotic use) are reported; these infections were not epidemiologically related. Yeast isolates were recovered from blood, ocular secretion, and bronchoalveolar lavage and were misidentified as C. catenulata and Candida albicans by the phenotypic MicroScan method. The isolates were confirmed to be C. auris by means of MALDI-TOF MS and DNA sequence analysis. Antifungal susceptibility testing was performed on these C. auris isolates, which exhibited high minimum inhibitory concentrations to triazoles and amphotericin B. One patient survived and the other two died. Only one of these deaths was related to fungemia. CONCLUSIONS: C. auris is an emerging and opportunistic multidrug-resistant human pathogen. It is necessary to strengthen measures to achieve an accurate and quick identification and also to avoid its dissemination. This will require improvements in health and infection control measures, as well as the promotion of antifungal stewardship in healthcare facilities.

Experiencia acerca del traslado de un hospital general y su repercusión en la actividad asistencial / Experience of the moving day of a general hospital and effect on clinical activity

Objetivos Describir el proceso del día del traslado de un hospital general. Analizar el impacto en la actividad asistencial en los primeros meses después del traslado. Método Hospital general de ámbito comarcal, en Cataluña, que se traslada a un nuevo edifico. Se describe el proceso del traslado de los pacientes y se analiza el impacto en la actividad (se comparan 3 meses a partir del traslado [período 2] con los mismos meses del año anterior [período1]).Resultados Se trasladaron 37 pacientes sin incidencias. Se atendieron 18.786 urgencias en el período 1 y 21.266 en el período 2 (+13,2%). El número de ingresos urgentes/programados fue de 1314/803 en el período 1 y de 1427/979 en el período 2 (+8,59%/+21,91%). Las visitas realizadas en consultas externas fueron 32.116 en el período 1 y 30.777 en el período 2 (-4,16%)Conclusiones Una adecuada planificación y coordinación permitió realizar un traslado de hospital sin incidencias y sin reducir la actividad asistencial(AU)

Objectives To describe the moving day of a general hospital and to analyze the impact on clinical activity in the first few months after the move. Methods We describe the process of patient transfer and analyze the impact on activity of moving a regional general hospital in Catalonia (Spain) to a new building. Activity was compared in two periods: the first 3 months after the move (period 2) and the same 3 months in the previous year (period 1).Results We moved 37 patients without incidents. A total of 18,786 patients were admitted to the emergency room in period 1 and 21,266 were admitted in period 2 (+13.2%). The number of admissions from the emergency room vs elective admissions was 1,314/803 in period 1 and 1,427/979 in period 2 (+8.59%/ +21.91%). The number of outpatient visits in period 1 was 32,116 and 30,777 in period 2 (-4.16%).Conclusions Proper planning and coordination allowed the hospital to be moved without incidents and without decreasing activity(SU)

[Experience of the moving day of a general hospital and effect on clinical activity]. / Experiencia acerca del traslado de un hospital general y su repercusión en la actividad asistencial.

OBJECTIVES: To describe the moving day of a general hospital and to analyze the impact on clinical activity in the first few months after the move. METHODS: We describe the process of patient transfer and analyze the impact on activity of moving a regional general hospital in Catalonia (Spain) to a new building. Activity was compared in two periods: the first 3 months after the move (period 2) and the same 3 months in the previous year (period 1). RESULTS: We moved 37 patients without incidents. A total of 18,786 patients were admitted to the emergency room in period 1 and 21,266 were admitted in period 2 (+13.2%). The number of admissions from the emergency room vs elective admissions was 1,314/803 in period 1 and 1,427/979 in period 2 (+8.59%/ +21.91%). The number of outpatient visits in period 1 was 32,116 and 30,777 in period 2 (-4.16%). CONCLUSIONS: Proper planning and coordination allowed the hospital to be moved without incidents and without decreasing activity.

Neogenin-mediated hemojuvelin shedding occurs after hemojuvelin traffics to the plasma membrane.

HFE2 (hemochromatosis type 2 gene) is highly expressed in skeletal muscle and liver hepatocytes. Its encoded protein, hemojuvelin (HJV), is a co-receptor for the bone morphogenetic proteins 2 and 4 (BMP2 and BMP4) and enhances the BMP-induced hepcidin expression. Hepcidin is a central iron regulatory hormone predominantly secreted from hepatocytes. HJV also binds neogenin, a membrane protein widely expressed in many tissues. Neogenin is required for the processing and release of HJV from cells. The role that neogenin plays in HJV trafficking was investigated, using HepG2 cells, a human hepatoma cell line. Knockdown of endogenous neogenin markedly suppresses HJV release but has no evident effect on HJV trafficking to the plasma membrane. The addition of a soluble neogenin ectodomain to cells markedly inhibits HJV release, indicating that the HJV shedding is not processed before trafficking to the cell surface. At the plasma membrane it undergoes endocytosis in a dynamin-independent but cholesterol-dependent manner. The additional findings that HJV release is coupled to lysosomal degradation of neogenin and that cholesterol depletion by filipin blocks both HJV endocytosis and HJV release suggest that neogenin-mediated HJV release occurs after the HJV-neogenin complex is internalized from the cell surface.

Evidence that inhibition of hemojuvelin shedding in response to iron is mediated through neogenin.

Hemojuvelin (HJV), encoded by the gene HFE2, is a critical upstream regulator of hepcidin expression. Hepcidin, the central iron regulatory hormone, is secreted from hepatocytes, whereas HFE2 is highly expressed in skeletal muscle and liver. Previous studies demonstrated that HJV is a GPI-anchored protein, binds the proteins neogenin and bone morphogenetic proteins (BMP2 and BMP4), and can be released from the cell membrane (shedding). In this study, we investigated the physiological significance and the underlying mechanism of HJV shedding. In acutely iron-deficient rats with markedly suppressed hepatic hepcidin expression, we detected an early phase increase of serum HJV with no significant change of either HFE2 mRNA or protein levels in gastrocnemius muscle. Studies in both C2C12 (a mouse myoblast cell line) and HepG2 (a human hepatoma cell line) cells showed active HJV shedding, implying that both skeletal muscle and liver could be the source of serum HJV. In agreement with the observations in iron-deficient rats, HJV shedding in these cell lines was down-regulated by holo-transferrin in a concentration-dependent manner. Our present study showing that knock-down of endogenous neogenin, a HJV receptor, in C2C12 cells suppresses HJV shedding and that overexpression of neogenin in HEK293 cells markedly enhances this process, suggests that membrane HJV shedding is mediated by neogenin. The finding that neither BMP4 nor its antagonist, noggin, was able to alter HJV shedding support the lack of involvement of BMP signaling pathway in this process.