Preparation, Characterization and Biological Studies of B-TCP and B-TCP/Al2O3 Scaffolds Obtained by Gel-Casting of Foams

Replacement tissues for tissue engineering can be produced by seeding human cells onto scaffolds. In order to guarantee adequate bio-compatibility, porosity and mechanical resistance for promoting cellular growth, proliferation and differentiation within scaffold structures, it is necessary to investigate and improve materials and processing routes. beta-tricalcium phosphate can be considered a very suitable bio-ceramic material for bone therapy because of its biocompatibility, osteo-conductivity and neo-vascularization potential. Alumina is commonly used as a sintering additive. In this study, beta-TCP and beta-TCP/Al2O3 scaffolds were obtained by gel-casting method. The scaffolds showed high porosity (8688%) and pore sizes ranging from 200 to 500 mu m. Even though alumina did not promote improvement in beta-TCP/Al2O3 scaffolds in terms of mechanical performance, they showed great cytocompatibility as there was no cytotoxic and genotoxic effect. Therefore, beta-TCP and beta-TCP/Al2O3 scaffolds are good candidates for application in tissue engineering.Sao Paulo Research Foundation - FAPESPNational Council for Scientific and Technological DevelopmentNational Council for Scientific and Technological DevelopmentUniv Fed Sao Paulo UNIFESP, Sci & Technol Inst ICT, Bioceram Lab BIOCERAM, 330 Talim St, BR-12231280 Sao Jose Dos Campos, SP, BrazilCNPEM, Brazilian Nanotechnol Natl Lab LNNANO, POB 6192, BR-13083970 Campinas, SP, BrazilUniv Estadual Paulista Julio de Mesquita Filho UN, Sch Dent, Dept Biosci & Oral Diag, 777 Engenheiro Francisco Jose Longo Ave, BR-12245000 Sao Jose Dos Campos, SP, BrazilUniv Fed Sao Paulo UNIFESP, Sci & Technol Inst ICT, Bioceram Lab BIOCERAM, 330 Talim St, BR-12231280 Sao Jose Dos Campos, SP, BrazilFAPESP: 2010/00863-0National Council for Scientific and Technological Development: 456461/2014-0]CNPq/PIBITI/UNIFESPWeb of Scienc

Human GLB1 knockout cerebral organoids: A model system for testing AAV9-mediated GLB1 gene therapy for reducing GM1 ganglioside storage in GM1 gangliosidosis

GM1 gangliosidosis is an autosomal recessive neurodegenerative disorder caused by the deficiency of lysosomal gangliosidebeta-galactosidase (beta-gal) and resulting in accumulation of GM1 ganglioside. The disease spectrum ranges from infantile to late onset and is uniformly fatal, with no effective therapy currently available. Although animal models have been useful for understanding disease pathogenesis and exploring therapeutic targets, no relevant human central nervous system (CNS) model system has been available to study its early pathogenic events or test therapies. To develop a model of human GM1 gangliosidosis in the CNS, we employed CRISPR/Cas9 genome editing to target GLB1 exons 2 and 6, common sites for mutations in patients, to create isogenic induced pluripotent stem (iPS) cell lines with lysosomal beta-gal deficiency. We screened for clones with \u3c 5% of parental cell line beta-gal enzyme activity and confirmed GLB1 knockout clones using DNA sequencing. We then generated GLB1 knockout cerebral organoids from one of these GLB1 knockout iPS cell clones. Analysis of GLB1 knockout organoids in culture revealed progressive accumulation of GM1 ganglioside. GLB1 knockout organoids microinjected with AAV9-GLB1 vector showed a significant increase in beta-gal activity and a significant reduction in GM1 ganglioside content compared with AAV9-GFP-injected organoids, demonstrating the efficacy of an AAV9 gene therapy-based approach in GM1 gangliosidosis. This proof-of-concept in a human cerebral organoid model completes the pre-clinical studies to advance to clinical trials using the AAV9-GLB1 vector

Cerebral organoids derived from Sandhoff disease-induced pluripotent stem cells exhibit impaired neurodifferentiation

Sandhoff disease, one of the GM2 gangliosidoses, is a lysosomal storage disorder characterized by the absence of beta-hexosaminidase A and B activity and the concomitant lysosomal accumulation of its substrate, GM2 ganglioside. It features catastrophic neurodegeneration and death in early childhood. How the lysosomal accumulation of ganglioside might affect the early development of the nervous system is not understood. Recently, cerebral organoids derived from induced pluripotent stem (iPS) cells have illuminated early developmental events altered by disease processes. To develop an early neurodevelopmental model of Sandhoff disease, we first generated iPS cells from the fibroblasts of an infantile Sandhoff disease patient, then corrected one of the mutant HEXB alleles in those iPS cells using CRISPR/Cas9 genome-editing technology, thereby creating isogenic controls. Next, we used the parental Sandhoff disease iPS cells and isogenic HEXB-corrected iPS cell clones to generate cerebral organoids that modeled the first trimester of neurodevelopment. The Sandhoff disease organoids, but not the HEXB-corrected organoids, accumulated GM2 ganglioside and exhibited increased size and cellular proliferation compared with the HEXB-corrected organoids. Whole-transcriptome analysis demonstrated that development was impaired in the Sandhoff disease organoids, suggesting that alterations in neuronal differentiation may occur during early development in the GM2 gangliosidoses

Warsaw Breakage Syndrome associated DDX11 helicase resolves G-quadruplex structures to support sister chromatid cohesion

Warsaw Breakage Syndrome (WABS) is a rare disorder related to cohesinopathies and Fanconi anemia, caused by bi-allelic mutations in DDX11. Here, we report multiple compound heterozygous WABS cases, each displaying destabilized DDX11 protein and residual DDX11 function at the cellular level. Patient-derived cell lines exhibit sensitivity to topoisomerase and PARP inhibitors, defective sister chromatid cohesion and reduced DNA replication fork speed. Deleting DDX11 in RPE1-TERT cells inhibits proliferation and survival in a TP53-dependent manner and causes chromosome breaks and cohesion defects, independent of the expressed pseudogene DDX12p. Importantly, G-quadruplex (G4) stabilizing compounds induce chromosome breaks and cohesion defects which are strongly aggravated by inactivation of DDX11 but not FANCJ. The DNA helicase domain of DDX11 is essential for sister chromatid cohesion and resistance to G4 stabilizers. We propose that DDX11 is a DNA helicase protecting against G4 induced double-stranded breaks and concomitant loss of cohesion, possibly at DNA replication forks

Quilombolas: a produção de mel na apicultura familiar do Vale do Ribeira, São Paulo

Family beekeeping (small-scale beekeeping) is an essential activity for economic and ecological farming systems (with its contribution to pollination), which aims to bring social inclusion, fixed income, and sustainable ecological bases. This study aimed to identify the current profile of honey production in five quilombolas communities in the São Paulo Ribeira Valley (Cangume, Pilões, Piririca, Porto Velho, and Ribeirão Grande/Terra Seca), with reference to the social and environmental aspects as well as beekeeping practices. The results suggest that the social aspects of family beekeeping can be considered a second income. In these communities, however, sanitation problems, including lack of potable water resources and mining contamination, differ between communities. With the study of georeferencing, we were able to establish local environmental conditions, which served as the basis for the floristic inventory of 85 plant species distributed in 32 botanical families. The analyses of air temperature and relative humidity and honey humidity indicated that the honey was influenced by climatic conditions and the processes adopted, since the measures of pH and EC suggest a correlation with the botanical origin of honeys. Professional training in maroon communities allowed an awareness of the quality and composition of honey.A apicultura familiar é uma atividade econômica e ecológica (devido à polinização) indispensável para sistemas de agricultura familiar que tem por finalidade trazer a inclusão social, renda fixa e bases ecológicas sustentáveis. Este trabalho teve como objetivo identificar o perfil atual da produção de mel de cinco comunidades quilombolas do Vale do Ribeira, São Paulo (Cangume, Pilões, Piririca, Porto Velho e, Ribeirão Grande/Terra Seca), quanto aos aspectos sociais, ambientais e das práticas de apicultura. Os resultados sugerem que nos aspectos sociais a apicultura familiar pode ser considerada uma segunda renda, que tem ainda problemas de condições sanitárias que diferem entre as comunidades, como falta de recurso de água potável e contaminação por mineração. Com o estudo de georreferenciamento, pôde-se estabelecer as condições ambientais locais que serviram de embasamento para o levantamento florístico de 85 espécies de plantas, distribuído em 32 famílias botânicas. As análises de umidade relativa e temperatura do ar e umidade do mel indicaram que os méis tiveram uma influência das condições climáticas e dos processos adotados. Já as medidas de pH e Condutividade Elétrica sugerem uma correlação com a origem botânica dos méis. A capacitação profissional nas comunidades permitiu uma sensibilização sobre a qualidade e composição do mel

Quilombolas: a produção de mel na apicultura familiar do Vale do Ribeira, São Paulo

A apicultura familiar é uma atividade econômica e ecológica (devido à polinização) indispensável para sistemas de agricultura familiar que tem por finalidade trazer a inclusão social, renda fixa e bases ecológicas sustentáveis. Este trabalho teve como objetivo identificar o perfil atual da produção de mel de cinco comunidades quilombolas do Vale do Ribeira, São Paulo (Cangume, Pilões, Piririca, Porto Velho e, Ribeirão Grande/Terra Seca), quanto aos aspectos sociais, ambientais e das práticas de apicultura. Os resultados sugerem que nos aspectos sociais a apicultura familiar pode ser considerada uma segunda renda, que tem ainda problemas de condições sanitárias que diferem entre as comunidades, como falta de recurso de água potável e contaminação por mineração. Com o estudo de georreferenciamento, pôde-se estabelecer as condições ambientais locais que serviram de embasamento para o levantamento florístico de 85 espécies de plantas, distribuído em 32 famílias botânicas. As análises de umidade relativa e temperatura do ar e umidade do mel indicaram que os méis tiveram uma influência das condições climáticas e dos processos adotados. Já as medidas de pH e Condutividade Elétrica sugerem uma correlação com a origem botânica dos méis. A capacitação profissional nas comunidades permitiu uma sensibilização sobre a qualidade e composição do mel

Evaluation of appendicitis risk prediction models in adults with suspected appendicitis

Background Appendicitis is the most common general surgical emergency worldwide, but its diagnosis remains challenging. The aim of this study was to determine whether existing risk prediction models can reliably identify patients presenting to hospital in the UK with acute right iliac fossa (RIF) pain who are at low risk of appendicitis. Methods A systematic search was completed to identify all existing appendicitis risk prediction models. Models were validated using UK data from an international prospective cohort study that captured consecutive patients aged 16–45 years presenting to hospital with acute RIF in March to June 2017. The main outcome was best achievable model specificity (proportion of patients who did not have appendicitis correctly classified as low risk) whilst maintaining a failure rate below 5 per cent (proportion of patients identified as low risk who actually had appendicitis). Results Some 5345 patients across 154 UK hospitals were identified, of which two‐thirds (3613 of 5345, 67·6 per cent) were women. Women were more than twice as likely to undergo surgery with removal of a histologically normal appendix (272 of 964, 28·2 per cent) than men (120 of 993, 12·1 per cent) (relative risk 2·33, 95 per cent c.i. 1·92 to 2·84; P < 0·001). Of 15 validated risk prediction models, the Adult Appendicitis Score performed best (cut‐off score 8 or less, specificity 63·1 per cent, failure rate 3·7 per cent). The Appendicitis Inflammatory Response Score performed best for men (cut‐off score 2 or less, specificity 24·7 per cent, failure rate 2·4 per cent). Conclusion Women in the UK had a disproportionate risk of admission without surgical intervention and had high rates of normal appendicectomy. Risk prediction models to support shared decision‐making by identifying adults in the UK at low risk of appendicitis were identified

Morfologia de esporos de pteridófitas do Parque Estadual das Fontes do Ipiranga (São Paulo, Brasil). Família: 21-Tectariaceae

Foram estudados os esporos de sete espécies de pteridófitas pertencentes à família Tectariaceae no Parque Estadual das Fontes do Ipiranga. A família está representada na área por três gêneros: Ctenitis, Lastreopsis e Megalastrum. Para todos os táxons estudados são apresentadas descrições, ilustrações, observações, bem como chave para a identificação das espécies