Defining functional diversity for lignocellulose degradation in a microbial community using multi-omics studies

Abstract\ud \ud Background\ud Lignocellulose is one of the most abundant forms of fixed carbon in the biosphere. Current industrial approaches to the degradation of lignocellulose employ enzyme mixtures, usually from a single fungal species, which are only effective in hydrolyzing polysaccharides following biomass pre-treatments. While the enzymatic mechanisms of lignocellulose degradation have been characterized in detail in individual microbial species, the microbial communities that efficiently breakdown plant materials in nature are species rich and secrete a myriad of enzymes to perform “community-level” metabolism of lignocellulose. Single-species approaches are, therefore, likely to miss important aspects of lignocellulose degradation that will be central to optimizing commercial processes.\ud \ud \ud Results\ud Here, we investigated the microbial degradation of wheat straw in liquid cultures that had been inoculated with wheat straw compost. Samples taken at selected time points were subjected to multi-omics analysis with the aim of identifying new microbial mechanisms for lignocellulose degradation that could be applied in industrial pre-treatment of feedstocks. Phylogenetic composition of the community, based on sequenced bacterial and eukaryotic ribosomal genes, showed a gradual decrease in complexity and diversity over time due to microbial enrichment. Taxonomic affiliation of bacterial species showed dominance of Bacteroidetes and Proteobacteria and high relative abundance of genera Asticcacaulis, Leadbetterella and Truepera. The eukaryotic members of the community were enriched in peritrich ciliates from genus Telotrochidium that thrived in the liquid cultures compared to fungal species that were present in low abundance. A targeted metasecretome approach combined with metatranscriptomics analysis, identified 1127 proteins and showed the presence of numerous carbohydrate-active enzymes extracted from the biomass-bound fractions and from the culture supernatant. This revealed a wide array of hydrolytic cellulases, hemicellulases and carbohydrate-binding modules involved in lignocellulose degradation. The expression of these activities correlated to the changes in the biomass composition observed by FTIR and ssNMR measurements.\ud \ud \ud Conclusions\ud A combination of mass spectrometry-based proteomics coupled with metatranscriptomics has enabled the identification of a large number of lignocellulose degrading enzymes that can now be further explored for the development of improved enzyme cocktails for the treatment of plant-based feedstocks. In addition to the expected carbohydrate-active enzymes, our studies reveal a large number of unknown proteins, some of which may play a crucial role in community-based lignocellulose degradation.This work was funded by Biotechnology and Biological Sciences Research\ud Council (BBSRC) Grants BB/1018492/1, BB/K020358/1 and BB/P027717/1, the\ud BBSRC Network in Biotechnology and Bioenergy BIOCATNET and São Paulo\ud Research Foundation (FAPESP) Grant 10/52362-5. ERdA thanks EMBRAPA\ud Instrumentation São Carlos and Dr. Luiz Alberto Colnago for providing the\ud NMR facility and CNPq Grant 312852/2014-2. The authors would like to thank\ud Deborah Rathbone and Susan Heywood from the Biorenewables Develop‑\ud ment Centre for technical assistance in rRNA amplicon sequencing

Bcl-2 Inhibits the Innate Immune Response during Early Pathogenesis of Murine Congenital Muscular Dystrophy

Laminin α2 (LAMA2)-deficient congenital muscular dystrophy is a severe, early-onset disease caused by abnormal levels of laminin 211 in the basal lamina leading to muscle weakness, transient inflammation, muscle degeneration and impaired mobility. In a Lama2-deficient mouse model for this disease, animal survival is improved by muscle-specific expression of the apoptosis inhibitor Bcl-2, conferred by a MyoD-hBcl-2 transgene. Here we investigated early disease stages in this model to determine initial pathological events and effects of Bcl-2 on their progression. Using quantitative immunohistological and mRNA analyses we show that inflammation occurs very early in Lama2-deficient muscle, some aspects of which are reduced or delayed by the MyoD-hBcl-2 transgene. mRNAs for innate immune response regulators, including multiple Toll-like receptors (TLRs) and the inflammasome component NLRP3, are elevated in diseased muscle compared with age-matched controls expressing Lama2. MyoD-hBcl-2 inhibits induction of TLR4, TLR6, TLR7, TLR8 and TLR9 in Lama2-deficient muscle compared with non-transgenic controls, and leads to reduced infiltration of eosinophils, which are key death effector cells. This congenital disease model provides a new paradigm for investigating cell death mechanisms during early stages of pathogenesis, demonstrating that interactions exist between Bcl-2, a multifunctional regulator of cell survival, and the innate immune response

Absenteísmo por doença ocupacional de trabalhadores rurais no setor canavieiro

The aim of this study was to analyze the frequency of work-related sick leave in sugarcane workers. A total of 1,230 medical excuses for 400 sugarcane workers were analyzed according to the International Classification of Diseases, 10th Revision (ICD-10). The following items were analyzed: diagnosis, sex, and length and season of sick leave. In all the seasons, musculoskeletal diseases showed the highest sick leave rate, following by respiratory diseases (p < 0.05). Sick leave due to musculoskeletal diseases was more frequent at the end of the sugarcane harvest than during the intercrop season (p < 0.05). Seventy-five percent of medical excuses were for one day. The longest sick leaves were for diseases of the genitourinary tract (p < 0.001). It is thus essential to have a multidisciplinary health team focused on the development of ergonomic activities, personal protective equipment, and health orientation specifically targeting this group’s needs.O objetivo do estudo foi analisar a frequência de afastamentos por doenças ocupacionais de trabalhadores rurais em uma safra canavieira. Foram analisados 1.230 atestados médicos classificados de acordo com a 10a revisão da Classificação Internacional de Doenças (CID-10) de 400 trabalhadores rurais. Nos atestados foram analisados: diagnóstico médico, sexo, dias de afastamento e período do afastamento. Em todos os períodos as doenças osteomusculares apresentaram maior frequência de atestados seguidos das doenças respiratórias (p < 0,05). Os atestados por doenças osteomusculares foram mais frequentes no fim da safra do que na entressafra (p < 0,05). Em relação ao número de dias afastados, 75% dos atestados foram de um dia. Os atestados por doenças do aparelho geniturinário obtiveram maior número de dias afastados (p < 0,001). Assim, é fundamental a atuação de uma equipe multidisciplinar de profissionais da saúde que atentem para o desenvolvimento de atividades de ergonomia, equipamentos de proteção individual e orientações de saúde direcionadas especificamente às necessidades dessa população.El objetivo del estudio fue analizar la frecuencia de ausencias por enfermedades ocupacionales de trabajadores rurales en una zafra azucarera. Fueron analizados 1.230 certificados médicos de acuerdo con la 10a revisión de la Classificación Internacional de las Enfermedades (CID-10) de 400 trabajadores rurales. En los certificados se examinaron: diagnóstico médico, sexo, días de ausencia y período de ausencia. En los períodos descritos las enfermedades del aparato locomotor mostraron una mayor frecuencia en los certificados, seguidas de las enfermedades respiratorias (p < 0,05). Los certificados por enfermedades del aparato locomotor fueron más frecuentes al final que en al principio de la zafra (p < 0,05). En relación con el número de días de ausencias, un 75% de los certificados tenían un día. Los certificados médicos de las enfermedades del sistema genitourinario tenían un mayor número de días de ausencias (p < 0,001). Por ello, es esencial trabajar con un equipo multidisciplinar de profesionales de la salud que se centren en el desarrollo de actividades de ergonomía, equipo de protección individual y orientación, dirigidos específicamente a las necesidades de esta población.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Estadual Paulista Júlio de Mesquita Filho Faculdade de Ciências e TecnologiaUniversidade Estadual Paulista Júlio de Mesquita Filho Faculdade de Ciências e Tecnologi