Nodulation in the absence of nod genes induction: alternative mechanisms involved in the symbiotic interaction between Cupriavidus sp. UYMMa02A and Mimosa pudica

Cupriavidus sp. UYMMa02A is a beta-rhizobia strain of the Cupriavidus genus, isolated from nodules of Mimosa magentea in Uruguay. This strain can form effective nodules with several Mimosa species, including its original host. Genome analyses indicate that Cupriavidus sp. UYMMa02A has a highly conserved 35 kb symbiotic island containing nod, nif, and fix operons, suggesting conserved mechanisms for the symbiotic interaction with plant hosts. However, while Cupriavidus sp. UYMMa02A produces functional nodules and promotes Mimosa pudica growth under nitrogen-limiting conditions, nod genes are not induced by luteolin or exposure to Mimosa spp. root exudate. To explore alternative mechanisms implicated in the Cupriavidus-Mimosa interaction, we assessed the proteomic profiles of Cupriavidus sp. UYMMa02A grown in the presence of pure flavonoids and co-culture with M. pudica plants. This approach allowed us to identify 24 differentially expressed proteins potentially involved in bacterial-plant interaction. In light of the obtained results, a possible model for nod-alternative symbiotic interaction is proposed.Agencia Nacional de Investigación e InnovaciónInstituto de Investigaciones Biológicas Clemente EstableProgama del Desarrollo de Ciencias Básica

Informe final del proyecto: Obtención de una cepa de Saccharomyces cerevisiae productora de una beta-glucosidasa de Issatchenkia terricola y explotación del genoma de esta levadura nativa para la identificación de nuevas enzimas con potencial aplicación en enología

El desarrollo del aroma del vino depende en gran parte de la existencia durante la elaboración, de enzimas (glicosidasas) capaces de actuar eficientemente sobre los sustratos glicosídicos existentes, generando compuestos volátiles. Estudios previos de nuestro grupo con enzimas aisladas de la microbiota de viñedos uruguayos, demostraron que una beta-glucosidasa de la cepa Issatchenkia terrícola presenta propiedades muy promisorias en condiciones enológicas y se destaca por impartir características aromáticas propias a los vinos locales. Sin embargo, los bajos niveles producidos por la cepa autóctona constituyen una limitante para la manipulación y posible aplicación biotecnológica de dicha glucosidasa. Con el objetivo de clonarla y expresarla en Saccharomyces cerevsiae con mayor rendimiento, nos encontramos actualmente focalizados en obtener la secuencia de dicha glucosidasa mediante el diseño de cebadores degenerados, dado que no disponemos aún del genoma de I. terrícola. La cepa generada será utilizada en ensayos de microvinificaciones y análisis químico y sensorial de aromas de los vinos obtenidos. Complementariamente, se propone avanzar en la caracterización molecular mediante secuenciación masiva del genoma de I. terrícola. Esto permitirá identificar la presencia de otros genes codificantes para beta-glucosidasas así como otras glicosidasas. Asimismo, la interpretación del genoma permitirá identificar otras actividades enzimáticas con potencial interés biotecnológico. El proyecto implica el diseño y uso racional del potencial existente en la microbiota nativa enológica, integrando conocimientos desde un enfoque multidisciplinario desde las áreas de bioquímica, biología molecular, genómica y química de aromas. Los resultados podrían generar productos potencialmente transferibles a la industria enológica.Agencia Nacional de Investigación e Innovació

SARS-CoV-2 susceptibility and COVID-19 disease severity are associated with genetic variants affecting gene expression in a variety of tissues

Variability in SARS-CoV-2 susceptibility and COVID-19 disease severity between individuals is partly due to genetic factors. Here, we identify 4 genomic loci with suggestive associations for SARS-CoV-2 susceptibility and 19 for COVID-19 disease severity. Four of these 23 loci likely have an ethnicity-specific component. Genome-wide association study (GWAS) signals in 11 loci colocalize with expression quantitative trait loci (eQTLs) associated with the expression of 20 genes in 62 tissues/cell types (range: 1:43 tissues/gene), including lung, brain, heart, muscle, and skin as well as the digestive system and immune system. We perform genetic fine mapping to compute 99% credible SNP sets, which identify 10 GWAS loci that have eight or fewer SNPs in the credible set, including three loci with one single likely causal SNP. Our study suggests that the diverse symptoms and disease severity of COVID-19 observed between individuals is associated with variants across the genome, affecting gene expression levels in a wide variety of tissue types

A first update on mapping the human genetic architecture of COVID-19

peer reviewe

ADVANTAGES AND PITFALLS OF ELECTRODIAGNOSTIC TESTS IN THE ASSESSMENT OF CHILDREN WITH NEUROMUSCULAR DISORDERS AND CORRELATION WITH THE MYOPATHOLOGICAL STUDY

Introduction: Neurophysiological investigations are an essential part in the diagnostic algorithm of neuromuscular pathologies, especially for patients with more heterogeneous phenotypes for which recognizing the early and subtle changes is essential. In children these techniques are often avoided because they are considered invasive, difficult to perform due to poor compliance, and difficult to interpret. Normative data of nerve conduction studies (NCSs) and electromyography (EMG) for the study of neuromuscular diseases in paediatric age are currently lacking in the literature. This greatly limits their clinical and research use. The diagnosis ultimately requires a muscle biopsy, regardless of the electrodiagnostic study results. Muscle biopsy has historically been considered to be the gold standard in the diagnosis of muscle disease. However, muscle biopsy is an expensive, invasive, time-consuming, and resource-dependent procedure. Aims and Methods: The first aim of this study was to collect and define a reference range of normal sensory and motor nerve conduction values of the upper and lower limbs during the maturation phases from newborn to three years of age and to collect and define a reference range of normal EMG values the maturation phases from newborn to six years of age, using quantitative EMG in the paediatric field to obtain the normative values of the main parameters, by means of the e-norms method. The secondary aim was to describe the diagnostic accuracy of electrodiagnostic tests (NCS and EMG) and muscle biopsy in children with suspected neuromuscular disorders. The study aimed to compare the sensitivity, specificity, positive and negative predictive value of the two tests and demonstrate in a systematic way if correlations between electrophysiological parameters and histopathologic findings in the same paediatric population exist; to performed the correlation between EMG and muscle biopsy variables Fisher’s exact test was used. Results: E-norms were calculated for Sural, Medial Plantar and Median sensory studies, and for Median, Peroneal and posterior Tibial motor studies. Regarding EMG, a reference range of motor unit potentials (MUPs) amplitude, MUPs duration, Turns/sec and Turn/Amplitude (TAA) was calculated. Regarding the diagnostic value of neurophysiological test and muscle biopsy, EMG was found to be particularly sensitive (100%) and specific (81%) in recognising neurogenic disorders, while muscle biopsy had a modest sensitivity (62%) and high specificity (89%). In our sample, the sensitivity of EMG in detecting myopathies is very low (26%), while the specificity remained high (95%); muscle biopsy had a very high diagnostic yield in myopathic disorders, with discrete sensitivity (65%), but very high specificity (100%). Some interesting correlations between EMG and histopathological results have been found. Increased amplitude of MUPs is correlated with the presence of fibre type grouping, while reduced amplitude is correlated with the presence of fibrosis and necrosis. Long duration of MUAPs is correlated with necrosis. Increased phases are sensitive for grouping and necrosis. A statistically significant correlation was also noted between a reduction of T/A at EMG and the presence of necrosis on muscle biopsy. Conclusions: Focused neurophysiological tests and muscle biopsy remain in children of all ages an indispensable diagnostic tool to guide clinicians in their decision-making and help in prompt diagnosis. The possibility of obtaining normal values from the neurophysiologist's laboratory adds their clinical and research use. Regarding EMG investigations, this study reports the first normative values for quantitative EMG in the international neurophysiological field, specifically the Turn/sec and TAA paediatric values. By demonstrating correlation between EMG findings and muscle biopsy results in a paediatric sample, this work could serve as a pilot study for future studies to better understand the muscle biopsy-neurophysiological tests correlation in specific neuromuscular disorders affecting developmental age

Introduction of NGS in environmental surveillance for Healthcare-associated infections control.

Introduction: Healthcare-associated infections (HAIs) and antibiotic resistance are the most frequent and serious cause of complications related to health care. The monitoring of hospital environmental surfaces takes place with cultural isolation with some performance limitations. Hence the urgency of implementing environmental surveillance systems using new generation methods. In this pilot study next generation sequencing (NGS) technologies were evaluated for hospital environmental surfaces microbiome characterization, compared with traditional and molecular PCR methods. Materials and Methods: Three randomized rooms were selected for two consecutive environmental sampling including floor, footboard and sink for a total of 36 critical points. NGS and comparison culture isolation and a custom molecular qPCR panel kit, were performed. Resistome, using a panel of 84 resistance genes, was additionally evaluated. Results: Pseudomonas, Acinetobacter, Streptococcus, Staphylococcus, Corynebacterium and Propionibacterium represented the most frequently bacterial area profile. Of them only Acinetobacter, Staphylococcus and Pseudomonas were isolated with cultural methods, Acinetobacter: 33/36 NGS vs 4/36; Staphylococcus:36/36 NGS vs 36/36, Pseudomonas: 34/36 NGS vs 1/36. Comparison with custom PCR analysis showed a rate of identification of Acinetobacter:10/36, Staphylococcus:36/36, and Pseudomonas: 29/36. Other microorganisms were detected from 1 to 29% for a total of 8 phyla and 58 different genera, showing the sink as the most representative site. To note, Streptococcus, detected by NGS in many samples was not sought with traditional methods. Basically resistome identified mainly genes conferring resistance to macrolides, aminoglycosides, fluoroquinolones and beta-lactams. Discussion and Conclusions: Data from this study proved as NGS can be an effective method in monitoring hospital indoor microbial communities. NGS data together with resistome, provide a deeper understanding of the development of HAIs leading to create new sanitation protocols

Informe final del proyecto: Obtención de una cepa de Saccharomyces cerevisiae productora de una beta-glucosidasa de Issatchenkia terricola y explotación del genoma de esta levadura nativa para la identificación de nuevas enzimas con potencial aplicación en enología

El desarrollo del aroma del vino depende en gran parte de la existencia durante la elaboración, de enzimas (glicosidasas) capaces de actuar eficientemente sobre los sustratos glicosídicos existentes, generando compuestos volátiles. Estudios previos de nuestro grupo con enzimas aisladas de la microbiota de viñedos uruguayos, demostraron que una beta-glucosidasa de la cepa Issatchenkia terrícola presenta propiedades muy promisorias en condiciones enológicas y se destaca por impartir características aromáticas propias a los vinos locales. Sin embargo, los bajos niveles producidos por la cepa autóctona constituyen una limitante para la manipulación y posible aplicación biotecnológica de dicha glucosidasa. Con el objetivo de clonarla y expresarla en Saccharomyces cerevsiae con mayor rendimiento, nos encontramos actualmente focalizados en obtener la secuencia de dicha glucosidasa mediante el diseño de cebadores degenerados, dado que no disponemos aún del genoma de I. terrícola. La cepa generada será utilizada en ensayos de microvinificaciones y análisis químico y sensorial de aromas de los vinos obtenidos. Complementariamente, se propone avanzar en la caracterización molecular mediante secuenciación masiva del genoma de I. terrícola. Esto permitirá identificar la presencia de otros genes codificantes para beta-glucosidasas así como otras glicosidasas. Asimismo, la interpretación del genoma permitirá identificar otras actividades enzimáticas con potencial interés biotecnológico. El proyecto implica el diseño y uso racional del potencial existente en la microbiota nativa enológica, integrando conocimientos desde un enfoque multidisciplinario desde las áreas de bioquímica, biología molecular, genómica y química de aromas. Los resultados podrían generar productos potencialmente transferibles a la industria enológica.Agencia Nacional de Investigación e Innovació

Predictors of survival in paediatric mitral valve replacement

The aim of this study was to identify the predictors of death and of reintervention after mitral valve replacement (MVR) in children