Axon micro-dissection and transcriptome profiling reveals the in vivo RNA content of fully differentiated myelinated motor axons

Axonal protein synthesis has been shown to play a role in developmental and regenerative growth, as well as in the maintenance of the axoplasm in steady state. Recent studies have begun to identify the mRNAs localized in axons, which could be translated locally under different conditions. Despite that now hundreds or thousands of mRNAs have been shown to be localized into the axonal compartment of cultured neurons in vitro, knowledge of which mRNAs are localized in mature myelinated axons is quite limited. With the purpose of characterizing the transcriptome of mature myelinated motor axons of peripheral nervous system, we modified the axon micro-dissection method devised by Koenig, enabling the isolation of the axoplasm RNA to perform RNA-seq analysis. The transcriptome analysis indicates that the number of RNAs detected in mature axons is lower in comparison with in vitro data, is depleted of glial markers and enriched in neuronal markers. The mature myelinated axons are enriched for mRNAs related to cytoskeleton, translation and oxidative phosphorylation. Moreover, it was possible to define core genes present in axons when comparing our data with transcriptomic data of axons grown in different conditions. This work provides evidence that axon micro-dissection is a valuable method to obtain data at genome-wide levels of mature and myelinated axons of the peripheral nervous system, and could be especially useful for the study of axonal involvement in neurodegenerative pathologies of motor neurons such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophies (SMA). Faria

Effectiveness of school-based nutrition education interventions to prevent and reduce excessive weight gain in children and adolescents: a systematic review

OBJETIVO: Avaliar a efetividade da educação nutricional em escolas na redução ou prevenção de sobrepeso e obesidade em crianças e adolescentes. FONTES DOS DADOS: Busca sistemática em 14 bases de dados e cinco revisões sistemáticas por ensaios controlados randomizados realizados em escolas com o objetivo de reduzir ou prevenir o sobrepeso em crianças e adolescentes. O índice de massa corporal e o consumo de frutas e verduras foram utilizados como desfechos primário e secundário, respectivamente. Não houve restrição de data de publicação ou idioma, exceto para idiomas com logogramas. Excluímos estudos em populações com distúrbios alimentares, dislipidemia, diabetes e deficiência física ou mental, e que utilizassem medicamentos ou suplementos alimentares. A avaliação por título e resumo e a avaliação de qualidade foram realizadas de forma independente por dois pesquisadores. Utilizamos as orientações do Centre for Reviews and Dissemination para a elaboração de artigos de revisão em saúde e o programa EPPI-Reviewer 3. SÍNTESE DOS DADOS: Das 4.809 referências inicialmente encontradas, 24 artigos preencheram os critérios de inclusão. Os dados extraídos apresentam evidências de efeitos antropométricos positivos e de aumento no consumo de frutas e verduras. As intervenções com as seguintes características demonstraram ser efetivas: duração > 1 ano, introdução como atividade regular da escola, envolvimento dos pais, introdução da educação nutricional no currículo regular e fornecimento de frutas e verduras pelos serviços de alimentação da escola. CONCLUSÃO: As intervenções para reduzir o sobrepeso e a obesidade e aumentar o consumo de frutas e verduras foram efetivas nos estudos bem conduzidos.OBJECTIVE: To evaluate the effectiveness of school-based nutrition education in reducing or preventing overweight and obesity in children and adolescents. SOURCES: Systematic search in 14 databases and five systematic reviews for randomized controlled trials conducted in schools to reduce or prevent overweight in children and adolescents. Body mass index and fruit and vegetable intake were used as primary and secondary measures of outcome, respectively. There was no restriction by date of publication or language, except for languages with structured logograms. We excluded studies on specific populations presenting eating disorders, dyslipidemia, diabetes, and physical or mental disabilities, as well as studies that used drugs or food supplements as components of the intervention. The assessment by title and abstract and the quality assessment were performed independently by two researchers. We used the Centre for Reviews and Dissemination's guidance for undertaking reviews in health care and the software EPPI-Reviewer 3. SUMMARY OF THE FINDINGS: From the initially retrieved 4,809 references, 24 articles met the inclusion criteria. The extracted data show that there is evidence of positive effects on anthropometry and of increase in fruit and vegetable consumption. Characteristics of the interventions that demonstrated effectiveness are: duration > 1 year, introduction into the regular activities of the school, parental involvement, introduction of nutrition education into the regular curriculum, and provision of fruits and vegetables by school food services. CONCLUSION: Interventions in schools to reduce overweight and obesity, as well as to increase fruits and vegetable consumption, have demonstrated effectiveness in the best-conducted studies

Evaluation of phosphorus levels in bones and serum of buffaloes (Bubalus bubalis) before and after supplementation with a selective mineral mixture

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação Amazônia de Amparo a Estudos e Pesquisas do Estado do Pará (FAPESPA), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) – Finance Code 001, Programa de Pós-graduação em Reprodução Animal na Amazônia (ReproAmazon), Instituto de Medicina Veterinária (IMV) and to Pró Reitora de Pesquisa e Pós-Graduação of the Universidade Federal do Pará (PROPESP-UFPA), Programa de Apoio à Publicação Qualificada - edital PAPQ/2022Universidade Federal do Pará-Campus Castanhal. Instituto de Medicina Veterinária. Castanhal, PA, Brazil.Universidade Federal do Sul e Sudeste do Pará. Instituto de Estudos do Trópico Úmido. Xinguara, PA, Brazil.Universidade Federal do Pará-Campus Castanhal. Instituto de Medicina Veterinária. Castanhal, PA, Brazil.Universidade Federal do Pará-Campus Castanhal. Instituto de Medicina Veterinária. Castanhal, PA, Brazil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Laboratório de Toxicologia. Ananindeua, PA, Brasil.Universidade Federal Rural da Amazônia. Instituto de Ciências Agrárias. Belém, PA, Brazil.Universidade Federal do Pará-Campus Castanhal. Instituto de Medicina Veterinária. Castanhal, PA, Brazil.Universidade Federal do Pará-Campus Castanhal. Instituto de Medicina Veterinária. Castanhal, PA, Brazil.This study aimed to evaluate phosphorus (P) concentration in serum and bone, the percentage of ash and the specific bone density of buffaloes on Ilha de Marajó before and after mineral supplementation. For this study, 14 crossbred buffaloes of Murrah and Mediterranean descent aged between 18 and 36 months were used. The average values of P before supplementation in serum and bone, the percentage of bone ash and the specific bone density were 5.68mg/dL±1.18, 16.53%±0.53, 59.95%±1.96 and 1.52g/cm3 ±0.32, respectively, which demonstrated P deficiency in animals raised on Ilha de Marajó. After supplementation with P for a period of seven months, the values were 6.61mg/dL±0.87, 16.90%±0.56 and 60.30%±0.95 and 1.71g/cm3 ±0.21, respectively. These results showed a significant increase in P concentration in blood serum, specific bone density and percentage of P in ash (P<0.05), but there was no significant increase in the percentage of ash. The average increase in P in the serum and ash did not reach normal levels in all animals; however, 28.6% of the animals had normal values of P in serum and 50% in the ash, and 64.3% had normal specific bone density values. The nonre-establishment, in some of the animals, of the variables of P serum and bone after supplementation for seven months may have occurred as a result of the low intake of the mineral mixture and by the low concentration of P in the Brachiaria brizantha cv. Marandu used for feeding animals during the experiment

Educating for earthquake science and risk in a tectonically slowly deforming region

Over the past decade, scientists have been called to participate more actively in public education and outreach (E&O). This is particularly true in fields of significant societal impact, such as earthquake science. Local earthquake risk culture plays a role in the way that the public engages in educational efforts. In this article, we describe an adapted E&O program for earthquake science and risk. The program is tailored for a region of slow tectonic deformation, where large earthquakes are extreme events that occur with long return periods. The adapted program has two main goals: (1) to increase the awareness and preparedness of the population to earthquake and related risks (tsunami, liquefaction, fires, etc.), and (2) to increase the quality of earthquake science education, so as to attract talented students to geosciences. Our integrated program relies on activities tuned for different population groups who have different interests and abilities, namely young children, teenagers, young adults, and professionals

Root Decomposition of Elephant Grass Pastures Grazed at Different Management Intensities

Grazing management may alter chemical composition of plant components affecting nutrient cycling. Among pasture management tools, adjustment of stocking rate (SR) and N fertilization have potential to affect nutrient cycling in the grassland ecosystem (Dubeux et al. 2007). Excreta from grazing animal and litter are the two major pathways of nutrient return on grazed pastures (Thomas 1992). Fertilization and SR may alter these pathways by different forms. Increasing fertilization generally increases pasture net primary productivity. Stocking rate affects different pasture and animal responses. Regarding nutrient cycling, increasing SR will likely increase proportion of nutrient returned through excreta as opposed to litter, increasing as a result nutrient losses (Dubeux et al. 2006). Root system may also be affected by management intensity. Frequent defoliation and low plant nutrition level may reduce root biomass (Richards 1993) and affect its decomposition. This study evaluated the effect of different SR´s and N fertilization levels on the decomposition of elephant grass roots

The usefulness of an Online Simplified Screening Questionnaire (SSQ) in identifying work-related cancers

To obtain a history of occupational exposure in the workplace, the questionnaire is one of the main sources of information. The aim of this study was to develop an online questionnaire using the REDCap data management platform based on the Work-Related Cancer Surveillance Guidelines, reported by the Brazilian National Cancer Institute. Several issues were taken into consideration for its routine application. It should be simple, easy, capable of being applied in a short time and used in the clinical setting of collecting information on the occupational history of the cancer patient. Consequently, this could enable the compulsory notification of work-related cancer. The questionnaire was developed based on questions about the use of and exposure to carcinogenic factors at work and due to smoking. An entirely electronic version of the cancer patient interview was performed using tablets. The online questionnaire was applied at the Barretos Cancer Hospital, Barretos, to newly diagnosed patients from July 2016 to 2018. A total of 1063 patients were included, and 550 indicated positively when asked “Do you work, or have you worked with this substance and/or in this function?/job?” Of these potentially notified patients, 38 subsequently had compulsorily reported work-related cancer. Another important result of this study was the creation and development of a website. In conclusion, we developed an online tool that could facilitate hospital routines, contributing to generating data for the compulsory notification of work-related cancer and triggering investigations and surveillance actions in Brazil.This research was funded by Barretos Cancer Hospital and the Public Ministry of Labor Campinas (Research, Prevention and Education of Occupational Cancer). Vazquez, F.L. is supported by the Public Ministry of Labor Campinas (Research, Prevention, and Education of Occupational Cancer) in Campinas, Brazil

The Trypanosoma cruzi nucleic acid binding protein Tc38 presents changes in the intramitochondrial distribution during the cell cycle

Background: Tc38 of Trypanosoma cruzi has been isolated as a single stranded DNA binding protein with high specificity for the poly [dT-dG] sequence. It is present only in Kinetoplastidae protozoa and its sequence lacks homology to known functional domains. Tc38 orthologues present in Trypanosoma brucei and Leishmania were proposed to participate in quite different cellular processes. To further understand the function of this protein in Trypanosoma cruzi, we examined its in vitro binding to biologically relevant [dT-dG] enriched sequences, its expression and subcellular localization during the cell cycle and through the parasite life stages.Results: By using specific antibodies, we found that Tc38 protein from epimastigote extracts participates in complexes with the poly [dT-dG] probe as well as with the universal minicircle sequence (UMS), a related repeated sequence found in maxicircle DNA, and the telomeric repeat. However, we found that Tc38 predominantly localizes into the mitochondrion. Though Tc38 is constitutively expressed through non-replicating and replicating life stages of T. cruzi, its subcellular localization in the unique parasite mitochondrion changes according to the cell cycle stage. in epimastigotes, Tc38 is found only in association with kDNA in G1 phase. From the S to G2 phase the protein localizes in two defined and connected spots flanking the kDNA. These spots disappear in late G2 turning into a diffuse dotted signal which extends beyond the kinetoplast. This later pattern is more evident in mitosis and cytokinesis. Finally, late in cytokinesis Tc38 reacquires its association with the kinetoplast. in non-replicating parasite stages such as trypomastigotes, the protein is found only surrounding the entire kinetoplast structure.Conclusions: the dynamics of Tc38 subcellular localization observed during the cell cycle and life stages support a major role for Tc38 related to kDNA replication and maintenance.FIRCAFondo Clemente Estable (DICyT)FAPESConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)PROSULPEDECIBAAMSUD-PasteurFac Ciencias, Lab Interacc Mol, Montevideo, UruguayFac Med, Dept Genet, Montevideo, UruguayFac Ciencias, Dept Biol Celular & Mol, Montevideo, UruguayInst Invest Biol Clemente Estable Montevideo Urug, Dept Neurobiol Celular & Mol, Montevideo, UruguayUniv Nacl Gen San Martin, CONICET, INTECH, Inst Invest Biotecnol, Buenos Aires, DF, ArgentinaUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, São Paulo, BrazilSUNY Buffalo, Dept Microbiol & Immunol, Buffalo, NY 14260 USAInst Biol Mol Parana, Curitiba, Parana, BrazilUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, São Paulo, BrazilFIRCA: R03 TW05665-01Fondo Clemente Estable (DICyT): 7109Web of Scienc

Analysis of binding properties and specificity through identification of the interface forming residues (IFR) for serine proteases in silico docked to different inhibitors

<p>Abstract</p> <p>Background</p> <p>Enzymes belonging to the same super family of proteins in general operate on variety of substrates and are inhibited by wide selection of inhibitors. In this work our main objective was to expand the scope of studies that consider only the catalytic and binding pocket amino acids while analyzing enzyme specificity and instead, include a wider category which we have named the Interface Forming Residues (IFR). We were motivated to identify those amino acids with decreased accessibility to solvent after docking of different types of inhibitors to sub classes of serine proteases and then create a table (matrix) of all amino acid positions at the interface as well as their respective occupancies. Our goal is to establish a platform for analysis of the relationship between IFR characteristics and binding properties/specificity for bi-molecular complexes.</p> <p>Results</p> <p>We propose a novel method for describing binding properties and delineating serine proteases specificity by compiling an exhaustive table of interface forming residues (IFR) for serine proteases and their inhibitors. Currently, the Protein Data Bank (PDB) does not contain all the data that our analysis would require. Therefore, an <it>in silico </it>approach was designed for building corresponding complexes</p> <p>The IFRs are obtained by "rigid body docking" among 70 structurally aligned, sequence wise non-redundant, serine protease structures with 3 inhibitors: bovine pancreatic trypsin inhibitor (BPTI), ecotine and ovomucoid third domain inhibitor. The table (matrix) of all amino acid positions at the interface and their respective occupancy is created. We also developed a new computational protocol for predicting IFRs for those complexes which were not deciphered experimentally so far, achieving accuracy of at least 0.97.</p> <p>Conclusions</p> <p>The serine proteases interfaces prefer polar (including glycine) residues (with some exceptions). Charged residues were found to be uniquely prevalent at the interfaces between the "miscellaneous-virus" subfamily and the three inhibitors. This prompts speculation about how important this difference in IFR characteristics is for maintaining virulence of those organisms.</p> <p>Our work here provides a unique tool for both structure/function relationship analysis as well as a compilation of indicators detailing how the specificity of various serine proteases may have been achieved and/or could be altered. It also indicates that the interface forming residues which also determine specificity of serine protease subfamily can not be presented in a canonical way but rather as a matrix of alternative populations of amino acids occupying variety of IFR positions.</p