Hepatic damage caused by flaviviruses: A systematic review.

Flaviviruses infect arthropods and mammals and their pathologies are a considerable global health problem, affecting about 400 million people per year. The symptoms of these flaviviruses range from mild manifestations such as nausea, vomiting, and headache to more serious cases such as hemorrhage, meningitis, microcephaly, kidney, and liver failure. This review aims to compile the morphological changes that occur due to infections caused by dengue, yellow fever, and Zika viruses, as well as to describe possible mechanisms of action of such flaviviruses in the liver. PRISMA guidelines were used to search for studies associating flavivirus with liver disorders. Two independent reviewers selected the studies on PubMed/Medline, Web of Science, and Scopus search platforms. The SYRCLE software was used for the evaluation of the study's quality. Eighteen experimental articles were included. The experimental animals often used in experiments were monkeys (5 %), hamsters (10 %), chicken embryos (10 %), and mice (75 %). It is evident that there is a strong hepatic interaction with flaviviruses, and the main hepatic alterations found were steatosis, apoptosis, necrosis, hemorrhage, elevation of ALT and AST levels, and total bilirubin. Flavivirus infection, in general, trigger an upregulation of pro-inflammatory cytokines, leading to structural changes in mitochondria that activate cascades of cellular death and promote insulin resistance. The majority of the studies primarily focus on dengue and yellow fever viruses, while the findings related to Zika virus exposure are still relatively limited and require further investigation.

Chemical and ecotoxicological indicators for evaluating the treatment of coal mining wastes.

This paper consists of the evaluation in regards to the ecotoxicological effectiveness of a treatment applied to a coal mining waste. The treatment consisted of separating the particles based on gravimetric concentration in spirals, generating three fractions: heavy, intermediate, and light, with high, moderate, and low pyrite content, respectively. The intermediate fraction represents the larger disposal volume of the waste on soils. To evaluate the effectiveness of the treatment, metal determination and bioassays Eisenia andrei, Folsomia candida, Lactuca sativa, Daphnia similis, and Raphidocelis subcapitata were applied to the intermediary fraction. To evaluate the toxicity to aquatic organisms, elutriates were generated from the unprocessed waste and the intermediate fraction. The intermediate fraction showed a decrease of metal concentrations compared to the untreated waste. Metal concentrations in the intermediate fraction were below the Brazilian thresholds for soil quality. Avoidance bioassay with E. andrei and germination tests of L. sativa showed no significant effects. The bioassay with F. candida indicated a significant reduction in reproduction at the highest doses used (24% and 50%). Bioassays with D. similis and R. subcapitata revealed a reduction in toxicity of the intermediate fraction compared to the untreated waste. However, the toxicity levels of the intermediate fraction to aquatic organisms still require attention, especially in regards to pH that played a crucial role in the toxicity. Finally, the results suggest that the treatment performed on the coal waste was efficient, even though significant toxicity have still been detected in the treated waste and additional steps are still required for adequate final disposal.

Genipin and glutaraldehyde based laccase two-layers immobilization with improved properties: New biocatalysts with high potential for enzymatic removal of trace organic contaminants.

This research proposes the preparation of a two-layer laccase biocatalyst using genipin or/and glutaraldehyde as cross-linking agents. The multilayer biocatalysts were prepared using different combinations of genipin and glutaraldehyde in the individual preparation of the first and second laccase layers. First, chitosan was treated with genipin or glutaraldehyde, followed by the immobilization of the first laccase layer to form a single-layer biocatalyst. Then, the immobilized laccases were coated once again with genipin or glutaraldehyde, and a new laccase layer was immobilized onto the system, resulting in the final two-layer biocatalyst. Compared to the single-layer biocatalysts, catalytic activity increased 1.7- and 3.4-fold when glutaraldehyde coating was used to prepare the second laccase layer. However, adding a second layer did not always produce more active biocatalysts, since the two-layer biocatalysts prepared with genipin (GenLacGenLac and GluLacGenLac) presented a decrease in activity of 65% and 28%, respectively. However, these two-layer biocatalysts prepared with genipin maintained 100% of their initial activity after 5 cycles of ABTS oxidation. Nevertheless, the two-layer, genipin-coated biocatalyst resulted in a higher removal of trace organic contaminants, since it removed 100% of mefenamic acid and 66% of acetaminophen, compared with the glutaraldehyde-coated biocatalyst, which removed 20% of mefenamic acid, and 18% of acetaminophen.

The Osteogenic Assessment of Mineral Trioxide Aggregate-based Endodontic Sealers in an Organotypic Ex Vivo Bone Development Model.

INTRODUCTION: Mineral trioxide aggregate (MTA)-based sealers are endodontic materials with widespread success in distinct clinical applications, potentially embracing direct contact with the bone tissue. Bone response to these materials has been traditionally addressed in vitro. Nonetheless, translational data are limited by the absence of native cell-to-cell and cell-to-matrix interactions that hinder the representativeness of the analysis. Ex vivo organotypic systems, relying on the culture of explanted biological tissues, preserve the cell/tissue composition, reproducing the spatial and organizational in situ complexity. This study was grounded on an innovative research approach, relying on the assessment of an ex vivo organotypic bone tissue culture system to address the osteogenic response to 3 distinct MTA-based sealers. METHODS: Embryonic chick femurs were isolated and grown ex vivo for 11 days in the presence of MTA Plus (Avalon Biomed Inc, Bradenton, FL), ProRoot MTA (Dentsply Tulsa Dental, Hohnson City, Germany), Biodentine (Septodont, Saint Maurdes Fosses, France), or AH Plus (Dentsply Sirona, Konstanz, Germany); the latter was used as a control material. Femurs were characterized by histologic, histochemical, and histomorphometric analysis. Gene expression assessment of relevant osteogenic markers was conducted by quantitative polymerase chain reaction. RESULTS: All MTA-based sealers presented an enhanced osteogenic performance compared with AH Plus. Histochemical and histomorphometric analyses support the increased activation of the osteogenic program by MTA-based sealers, with enhanced collagenous matrix deposition and tissue mineralization. Gene expression analysis supported the enhanced activation of the osteogenic program. Comparatively, ProRoot MTA induced the highest osteogenic functionality on the characterized femurs. CONCLUSIONS: MTA-based sealers enhanced the osteogenic activity within the assayed organotypic bone model, which was found to be a sensitive system for the assessment of osteogenic modulation mediated by endodontic sealers.

No effect of prior Dengue virus 1 infection in mouse dams on long-term behavioral profiles in offspring infected with Zika virus during gestation.

Zika virus (ZIKV) is a mosquito-borne Flavivirus structurally and antigenically related to Dengue virus (DENV). Zika virus has been associated with congenital anomalies and most ZIKV outbreaks have occurred in endemic areas of DENV. The present study investigated the effects of prior DENV serotype 1 (DENV1) immunity in immunocompetent female Swiss mice on gestational ZIKV infection in offspring. Physical/reflex development, locomotor activity, anxiety, visual acuity, and brain-derived neurotrophic factor (BDNF) levels were evaluated in offspring during infancy and adolescence. Anti-DENV1 and anti-ZIKV antibodies were detected in sera of the progenitors, whereas no ZIKV genomes were detected in the offspring brain. Pups from dams with only DENV1 immunity presented alterations of physical/reflex development. Pups from all infected dams exhibited time-related impairments in locomotor activity and anxiolytic-like behavior. Offspring from DENV/ZIKV-infected dams exhibited impairments in visual acuity during infancy but not during adolescence, which was consistent with morphometric analysis of the optic nerve. Pups from DENV1-, ZIKV-, and DENV/ZIKV-infected dams exhibited a decrease in BDNF levels during infancy and an increase during adolescence in distinct brain regions. In summary, we found no influence of prior DENV1 immunity on gestational ZIKV infection in offspring, with the exception of alterations of early visual parameters, and an increase in BDNF levels in the hippocampus during adolescence.

Imbalanced cellular metabolism compromises cartilage homeostasis and joint function in a mouse model of mucolipidosis type III gamma.

Mucolipidosis type III (MLIII) gamma is a rare inherited lysosomal storage disorder caused by mutations in GNPTG encoding the γ-subunit of GlcNAc-1-phosphotransferase, the key enzyme ensuring proper intracellular location of multiple lysosomal enzymes. Patients with MLIII gamma typically present with osteoarthritis and joint stiffness, suggesting cartilage involvement. Using Gnptg knockout (Gnptgko ) mice as a model of the human disease, we showed that missorting of a number of lysosomal enzymes is associated with intracellular accumulation of chondroitin sulfate in Gnptgko chondrocytes and their impaired differentiation, as well as with altered microstructure of the cartilage extracellular matrix (ECM). We also demonstrated distinct functional and structural properties of the Achilles tendons isolated from Gnptgko and Gnptab knock-in (Gnptabki ) mice, the latter displaying a more severe phenotype resembling mucolipidosis type II (MLII) in humans. Together with comparative analyses of joint mobility in MLII and MLIII patients, these findings provide a basis for better understanding of the molecular reasons leading to joint pathology in these patients. Our data suggest that lack of GlcNAc-1-phosphotransferase activity due to defects in the γ-subunit causes structural changes within the ECM of connective and mechanosensitive tissues, such as cartilage and tendon, and eventually results in functional joint abnormalities typically observed in MLIII gamma patients. This idea was supported by a deficit of the limb motor function in Gnptgko mice challenged on a rotarod under fatigue-associated conditions, suggesting that the impaired motor performance of Gnptgko mice was caused by fatigue and/or pain at the joint.This article has an associated First Person interview with the first author of the paper.

Influence of maternal age and hospital characteristics on the mode of delivery.

OBJECTIVES: to analyze the relationship between maternal age and the source of healthcare payment with mode of delivery in public and private national hospitals between the years 2012 to 2017, and the length of hospital stay. METHODS: cross-sectional study of 91,894 women who had children in public and private hospitals between 2012 and 2017. Data were collected from the Diagnosis-Related Groups Brazil system and a comparative analysis was performed between patients in public care and those in supplementary healthcare. RESULTS: in public care, the majority were vaginal deliveries and the reverse occurred in supplementary health. The proportion of cesarean sections was higher in the age group 31 to 40 years old in both services. The hospital stay was longer among women who underwent a cesarean section. CONCLUSIONS: high maternal age and the source of healthcare payment influence the mode of delivery, which interfere with the length of hospital stay.

Echocardiographic study in children with osteogenesis imperfecta.

BACKGROUND: Osteogenesis imperfecta is a collagen type I bone disorder. Recently, extra-skeletal manifestations have been described, including many cardiovascular alterations. This study aims to report echocardiogram study in children with osteogenesis imperfecta compared to a control group. METHODS: A cross-sectional comparative study took place in the Reference Center for Treatment of Osteogenesis Imperfecta in Southern Brazil. Fifty-four patients with osteogenesis imperfecta were paired with 54 controls, based on body surface area, and echocardiogram findings were compared. RESULTS: All cases were asymptomatic for cardiac manifestations. The case group presented significant larger values in aortic diameter, left atrium diameter, left ventricule end-diastolic diameter, left ventricule end-systolic diameter, and right ventricle diameter compared with the control group. The analysis considering the severity of osteogenesis imperfecta shows that in mild osteogenesis imperfecta, the aortic diameter (p < 0.001), left atrium diameter (p = 0.002), left ventricule end-diastolic diameter (p = 0.001), left ventricule end-systolic diameter (p = 0.026), and right ventricle diameter (p < 0.001) were significantly larger than in the control group. Patients with moderate/severe osteogenesis imperfecta had similar results, with aortic diameter (p < 0.001), left atrium diameter (p < 0.001), left ventricule end-diastolic diameter (p = 0.013), and left ventricule end-systolic diameter (0.004) statistically larger than controls. Twenty-six (48.1%) of the cases had physiological tricuspid regurgitation and in controls this finding was observed in eight (14.8%) (p < 0.001). CONCLUSION: Children with osteogenesis imperfecta presented cardiac function within the normal pattern, but dimensions of left ventricular dimensions were increased compared to the ones of the controls.

PathMolD-AB: Spatiotemporal pathways of protein folding using parallel molecular dynamics with a coarse-grained model.

Solving the protein folding problem (PFP) is one of the grand challenges still open in computational biophysics. Globular proteins are believed to evolve from initial configurations through folding pathways connecting several thermodynamically accessible states in a free energy landscape until reaching its minimum, inhabited by the stable native structures. Despite its huge computational burden, molecular dynamics (MD) is the leading approach in the PFP studies by preserving the Newtonian temporal evolution in the canonical ensemble. Non-trivial improvements are provided by highly parallel implementations of MD in cost-effective GPUs, concomitant to multiscale descriptions of proteins by coarse-grained minimalist models. In this vein, we present the PathMolD-AB framework, a comprehensive software package for massively parallel MD simulations using the canonical ensemble, structural analysis, and visualization of the folding pathways using the minimalist AB-model. It has, also, a tool to compare the results with proteins re-scaled from the PDB. We simulate and analyze, as case studies, the folding of four proteins: 13FIBO, 2GB1, 1PLC and 5ANZ, with 13, 55, 99 and 223 amino acids, respectively. The datasets generated from simulations correspond to the MD evolution of 3500 folding pathways, encompassing 35×106 states, which contains the spatial amino acid positions, the protein free energies and radii of gyration at each time step. Results indicate that the speedup of our approach grows logarithmically with the protein length and, therefore, it is suited for most of the proteins in the PDB. The predicted structures simulated by PathMolD-AB were similar to the re-scaled biological structures, indicating that it is promising for the study of the PFP study.

Influence of maternal age and hospital characteristics on the mode of delivery / Influencia de la edad materna y las características del hospital en la ruta de nacimiento / Influência da idade materna e das características hospitalares nas vias de nascimento

ABSTRACT Objectives: to analyze the relationship between maternal age and the source of healthcare payment with mode of delivery in public and private national hospitals between the years 2012 to 2017, and the length of hospital stay. Methods: cross-sectional study of 91,894 women who had children in public and private hospitals between 2012 and 2017. Data were collected from the Diagnosis-Related Groups Brazil system and a comparative analysis was performed between patients in public care and those in supplementary healthcare. Results: in public care, the majority were vaginal deliveries and the reverse occurred in supplementary health. The proportion of cesarean sections was higher in the age group 31 to 40 years old in both services. The hospital stay was longer among women who underwent a cesarean section. Conclusions: high maternal age and the source of healthcare payment influence the mode of delivery, which interfere with the length of hospital stay.

RESUMEN Objetivos: analizar la relación entre la edad materna y la fuente de pago de la atención con las rutas de nacimiento en hospitales nacionales públicos y privados entre los años 2012 a 2017, y la duración de la estancia hospitalaria. Métodos: estudio transversal con 91.894 mujeres que tuvieron hijos en hospitales públicos y privados entre 2012 y 2017. Se recopilaron datos del sistema Diagnosis-Related Groups Brasil y se realizó un análisis comparativo entre pacientes en atención pública y aquellos en atención médica complementaria. Resultados: en la atención pública, la mayoría fueron partos vaginales y lo contrario ocurrió en la atención complementaria. La proporción de cesáreas fue mayor en el grupo de edad de 31 a 40 años en ambos servicios. La estadía en el hospital fue más larga entre las mujeres que se sometieron a una cesárea. Conclusiones: La edad materna elevada y la fuente de pago influyen en la ruta de nacimiento, lo que interfiere con la duración de la estancia hospitalaria.

RESUMO Objetivos: analisar a relação da idade materna e da fonte pagadora da assistência com as vias de nascimento em hospitais nacionais públicos e privados entre os anos de 2012 a 2017, e o tempo de permanência hospitalar. Métodos: estudo transversal com 91.894 mulheres que tiveram seus filhos em hospitais públicos e privados entre 2012 a 2017. Os dados foram coletados a partir do sistema Diagnosis-Related Groups Brasil e submetidos à análise comparativa entre pacientes do atendimento público e da saúde suplementar. Resultados: no atendimento público, a maioria dos nascimentos ocorreram por via vaginal e o inverso ocorreu na saúde suplementar. Observou-se que a proporção de cesariana foi maior na faixa etária de 31 a 40 anos, em ambos os serviços, e maior tempo de permanência hospitalar nas mulheres submetidas à cesariana. Conclusões: idade materna elevada e fonte de financiamento influenciam nas vias de nascimento, que interferem no tempo de permanência hospitalar.

Influence of maternal age and hospital characteristics on the mode of delivery / Influencia de la edad materna y las características del hospital en la ruta de nacimiento / Influência da idade materna e das características hospitalares nas vias de nascimento

ABSTRACT Objectives: to analyze the relationship between maternal age and the source of healthcare payment with mode of delivery in public and private national hospitals between the years 2012 to 2017, and the length of hospital stay. Methods: cross-sectional study of 91,894 women who had children in public and private hospitals between 2012 and 2017. Data were collected from the Diagnosis-Related Groups Brazil system and a comparative analysis was performed between patients in public care and those in supplementary healthcare. Results: in public care, the majority were vaginal deliveries and the reverse occurred in supplementary health. The proportion of cesarean sections was higher in the age group 31 to 40 years old in both services. The hospital stay was longer among women who underwent a cesarean section. Conclusions: high maternal age and the source of healthcare payment influence the mode of delivery, which interfere with the length of hospital stay.

RESUMEN Objetivos: analizar la relación entre la edad materna y la fuente de pago de la atención con las rutas de nacimiento en hospitales nacionales públicos y privados entre los años 2012 a 2017, y la duración de la estancia hospitalaria. Métodos: estudio transversal con 91.894 mujeres que tuvieron hijos en hospitales públicos y privados entre 2012 y 2017. Se recopilaron datos del sistema Diagnosis-Related Groups Brasil y se realizó un análisis comparativo entre pacientes en atención pública y aquellos en atención médica complementaria. Resultados: en la atención pública, la mayoría fueron partos vaginales y lo contrario ocurrió en la atención complementaria. La proporción de cesáreas fue mayor en el grupo de edad de 31 a 40 años en ambos servicios. La estadía en el hospital fue más larga entre las mujeres que se sometieron a una cesárea. Conclusiones: La edad materna elevada y la fuente de pago influyen en la ruta de nacimiento, lo que interfiere con la duración de la estancia hospitalaria.

RESUMO Objetivos: analisar a relação da idade materna e da fonte pagadora da assistência com as vias de nascimento em hospitais nacionais públicos e privados entre os anos de 2012 a 2017, e o tempo de permanência hospitalar. Métodos: estudo transversal com 91.894 mulheres que tiveram seus filhos em hospitais públicos e privados entre 2012 a 2017. Os dados foram coletados a partir do sistema Diagnosis-Related Groups Brasil e submetidos à análise comparativa entre pacientes do atendimento público e da saúde suplementar. Resultados: no atendimento público, a maioria dos nascimentos ocorreram por via vaginal e o inverso ocorreu na saúde suplementar. Observou-se que a proporção de cesariana foi maior na faixa etária de 31 a 40 anos, em ambos os serviços, e maior tempo de permanência hospitalar nas mulheres submetidas à cesariana. Conclusões: idade materna elevada e fonte de financiamento influenciam nas vias de nascimento, que interferem no tempo de permanência hospitalar.

Formação profissional em uma atividade vivencial em saúde mental: Grupo Comunitário de Saúde Mental / Professional education in an experiential activity in mental health: Mental Health Community Group / Formación Profesional en una actividad vivencial en salud mental: Grupo Comunitario de Salud Mental

Objetivou-se compreender as experiências de estudantes e profissionais em processo de formação com uma atividade vivencial em saúde mental, denominada Grupo Comunitário de Saúde Mental (GCSM) e os desdobramentos de sua apropriação no percurso de formação profissional. Adotou-se uma abordagem qualitativa, de natureza exploratória, tendo como instrumento a entrevista aberta. Participaram cinco estudantes e 10 profissionais em formação de diferentes áreas da saúde. Para a análise utilizou-se o método de Análise de Conteúdo Temática. Foram elaboradas duas categorias: O lugar profissional no GCSM; A horizontalidade das relações no GCSM. Considera-se que a proposta vivencial, presente no modelo do GCSM apresenta contribuições tanto do ponto de vista técnico da formação, quanto em relação ao desenvolvimento humano dos profissionais. Destaca-se que este estudo traz avanços para se pensar a formação realizada em serviços-escola no campo da saúde mental e que tal compreensão pode colaborar com a consolidação do modelo psicossocial de atenção.

The objective of this study was to understand the experiences of students and professionals in process of education in an experiential activity in mental health, named Mental Health Community Group (MHCG), and the unfolding of its appropriation throughout professional education. A qualitative exploratory approach was adopted, and open interviews were used as data collection instrument. Study participants were five students and 10 professionals studying in different healthcare areas. Analysis was conducted using the thematic content analysis technique. Two categories were created: The professional place in the MHCG, and The horizontality of relationships in the MHCG. The experiential proposal, in the MHCG model, presents contributions both from the technical point of view of education, and in relation to the human development of professionals. This study promotes reflection on the education provided in teaching services in the mental health area and this understanding can contribute to consolidate the psychosocial care model.

Se trató de comprender las experiencias de estudiantes y profesionales en proceso de formación con una actividad vivencial en salud mental, denominada Grupo Comunitario de Salud Mental (GCSM) y los desdoblamientos de su apropiación en el curso de formación profesional. Se adoptó un enfoque cualitativo, de naturaleza exploratoria, teniendo como instrumento la entrevista abierta. Participaron cinco estudiantes y 10 profesionales en formación de diferentes áreas de la salud. Para el análisis se utilizó el método de Análisis de Contenido Temático. Se elaboraron dos categorías: El lugar profesional en el GCSM y la horizontalidad de las relaciones en el GCSM. Se considera que la propuesta vivencial, presente en el modelo del GCSM ofrece contribuciones tanto desde el punto de vista técnico de la formación, como con relación al desarrollo humano de los profesionales. Se destaca que este estudio trae avances para pensar la formación realizada en servicios-escuela en el campo de la salud mental y que tal comprensión puede colaborar con la consolidación del modelo psicosocial de atención.

Immobilization of Lipase A from Candida antarctica onto Chitosan-Coated Magnetic Nanoparticles.

In this communication, lipase A from Candida antarctica (CALA) was immobilized by covalent bonding on magnetic nanoparticles coated with chitosan and activated with glutaraldehyde, labelled CALA-MNP, (immobilization parameters: 84.1% ± 1.0 for immobilization yield and 208.0 ± 3.0 U/g ± 1.1 for derivative activity). CALA-MNP biocatalyst was characterized by X-ray Powder Diffraction (XRPD), Fourier Transform Infrared (FTIR) spectroscopy, Thermogravimetry (TG) and Scanning Electron Microscope (SEM), proving the incorporation of magnetite and the immobilization of CALA in the chitosan matrix. Besides, the immobilized biocatalyst showed a half-life 8-11 times higher than that of the soluble enzyme at pH 5-9. CALA showed the highest activity at pH 7, while CALA-MNP presented the highest activity at pH 10. The immobilized enzyme was more active than the free enzyme at all studied pH values, except pH 7.

Cyclic pamidronate treatment for osteogenesis imperfecta: Report from a Brazilian reference center.

Treatment of moderate and severe forms of osteogenesis imperfecta (OI) with cyclic pamidronate at the Reference Center for OI Treatment in Southern Brazil was studied. A retrospective cohort study was conducted from 2002 to 2012. Data were obtained during inpatient (drug infusion) and outpatient care. Clinical data, including the presence of blue sclerae, dentinogenesis imperfecta, history and site of the fractures, biochemical data, including calcium, phosphorus, and alkaline phosphatase levels, were systematically collected. Bone mineral density (BMD) was measured using dual energy X-ray absorptiometry (DXA). Forty-five patients (26 females) were included in the study, and the age of the patients at the time of diagnosis ranged from 1 to 144 months, with a median age (p25-p75) of 38 (5-96) months. Most cases presented OI-4 (51.1%), and the median age of the patients at the start of treatment was 3.3 years (25-75 percentiles: 0.5 - 8.7 years). Twenty-four patients (54.5%) had some adverse events or intercurrences during treatment, and the treatment compliance mean was 92.3% (± 10.7). The treatment with intravenous pamidronate has shown to be safe, well-tolerated, and effective in regard to the improvement of BMD and the reduction of the number of fractures in children and adolescents with OI.

Chitosan activated with divinyl sulfone: a new heterofunctional support for enzyme immobilization. Application in the immobilization of lipase B from Candida antarctica.

A novel heterofunctional support for enzyme immobilization, chitosan-divinyl sulfone, was assessed in this study. The activation of chitosan with DVS was carried out at three different pHs (10.0, 12.5 and 14.0) and a Candida antarctica Lipase B (CALB) was selected as the model enzyme. After immobilization, the biocatalysts were incubated under alkaline conditions in a buffer to facilitate the multipoint covalent attachment, followed by incubation in ethylenediamine (EDA) aiming at blocking the remaining reactive groups. The highest thermal stability was obtained when pH 10.0 was used during support activation. These results were shown to be better than those obtained when using glutaraldehyde as the support-activating reagent. Subsequently, the immobilization pH was investigated (5.0, 7.0 and 10.0) prior to alkaline incubation, with the highest enzyme stability levels found at pH 10.0. Finally, the selected biocatalyst was used in the hydrolysis of ethyl hexanoate and presented an activity of 14,520.37 U/g of immobilized lipase at pH 5.0. These results show that chitosan activated with divinyl sulfone is a very promising support for enzyme immobilization and the proposed protocol is able to successfully improve enzyme stability.

Efficient biotechnological synthesis of flavor esters using a low-cost biocatalyst with immobilized Rhizomucor miehei lipase.

In this work, the synthesis of two fruit flavor esters, namely methyl and ethyl butyrate, by lipase from Rhizomucor miehei immobilized onto chitosan in the presence of the surfactant sodium dodecyl sulfate SDS was investigated. In the optimized conditions, maximum esterification yield for ethyl butyrate and methyl butyrate was (92 ± 1%) and (89 ± 1%), respectively. Esterification yields for both reactions were comparable or even superior to the ones achieved when the synthesis was catalyzed by a commercial enzyme, Lipozyme®, at the same reaction conditions. For ethyl butyrate, the developed biocatalyst was used for seven consecutive cycles of reaction with retention of its catalytic activity. For methyl butyrate synthesis the biocatalyst was used for four consecutive cycles without loss of its catalytic activity. The results show that chitosan may be employed in obtaining biocatalysts with high catalytic efficiency and can successfully replace the currently commercial available biocatalysts.

Moderate endemicity of the human T-lymphotropic virus infection in the metropolitan region of Belém, Pará, Brazil. / Moderada endemicidade da infecção pelo vírus linfotrópico-T humano na região metropolitana de Belém, Pará, Brasil.

INTRODUCTION: The spread of the HTLV infection in families living in the metropolitan area of Belém, Pará, Brazil, and the lack of studies in the general population requires studies to better understand its prevalence in the region. METHODS: An anti-HTLV-1/HTLV-2 antibodies test was carried out on random adults in public places in Belém between November 2014 and November 2015. A proviral DNA test detected if the person was infected, and then a clinical evaluation and an intrafamilial investigation were carried out. RESULTS: Of the 1059 individuals being investigated, 21 (2.0%) had seroreagent samples, 15 (1.4%) had HTLV-1, 5 (0.5%) had HTLV-2, and proviral DNA was undetectable in one case. The mean age of the infected people (57.2) was higher than that of those that were uninfected (46.2) (p = 0.0010). The prevalence of infection increased with age, especially in individuals with a family income equal to or less than a minimum wage. Intrafamilial transmission seems to have occurred in all of the families being studied. Among the patients with HTLV-1, 30% (3/10) already had some symptom related to the infection. DISCUSSION: The increase in prevalence rates according to age may be due to late seroconversion of a previously acquired infection, or the cumulative risk of new infections, especially in women. CONCLUSION: There was a moderate prevalence of the HTLV infection among adult individuals from the metropolitan area of Belém, with a predominance of HTLV-1. This infection was associated with low income and increasingly older women. It also presented intrafamily spread and negligence in the diagnosis of associated diseases.

INTRODUÇÃO: A disseminação da infecção pelo vírus linfotrópico-T humano (HTLV) em famílias da área metropolitana de Belém, Pará, Brasil, e a ausência de estudos na população em geral requisitam investigações que esclareçam melhor a sua prevalência na região. METODOLOGIA: Foi realizada pesquisa de anticorpos anti-HTLV-1/HTLV-2 em indivíduos adultos transeuntes de logradouros públicos de Belém, entre novembro de 2014 e novembro de 2015. A infecção foi confirmada por pesquisa de DNA proviral e foi realizada avaliação clínica e investigação intrafamiliar dos infectados. RESULTADOS: Dos 1.059 indivíduos investigados, 21 (2,0%) apresentaram amostras sororeagentes, 15 (1,4%) confirmados para HTLV-1, 5 (0,5%) para HTLV-2 e o DNA proviral foi indetectável em 1 caso. A média de idade dos infectados (57,2) foi maior que a dos não infectados (46,2) (p = 0,0010). A infecção aumentou com a idade e se destacou nos indivíduos com renda familiar menor ou igual a um salário mínimo. A transmissão intrafamiliar parece ter ocorrido em todas as famílias investigadas. Dentre os portadores de HTLV-1, 30% (3/10) já apresentavam algum sintoma relacionado à infecção. DISCUSSÃO: O aumento da infecção de acordo com a idade pode ocorrer por soroconversão tardia de infecção pré-adquirida ou pelo risco cumulativo de novas infecções, sobretudo em mulheres. CONCLUSÃO: A infecção por HTLV demonstrou moderada prevalência na população estudada, com predomínio do HTLV-1. Essa mostrou-se associada à baixa renda e ao aumento da idade das mulheres. Também apresentou disseminação intrafamiliar e negligência no diagnóstico das doenças associadas.

Stability of the HTLV-1 glycoprotein 46 (gp46) gene in an endemic region of the Brazilian Amazon and the presence of a significant mutation (N93D) in symptomatic patients.

BACKGROUND: The human T-lymphotropic virus type 1 (HTLV-1) affects 2-5 million people worldwide, and is associated with a number of degenerative and infectious diseases. The Envelope glycoproteins (gp) are highly conserved among the different HTLV-1 isolates, although nucleotide substitutions in the region that codifies these proteins may influence both the infectivity and the replication of the virus. The gp46 gene has functional domains which have been associated with the inhibition of the formation of the syncytium, cell-cell transmission, and the production of antibodies. The present study investigated the genetic stability of the gp46 gene of HTLV-1 in an endemic region of Brazilian Amazonia. METHODS: Index case (IC - a sample of a given family group) carriers of HTLV-1 were investigated in the metropolitan region of Belém (Pará, Brazil) between January 2010 (registered retrospectively) and December 2015. The sequences that codify the gp46 were amplified by PCR, purified and sequenced (MF084788-MF084825). The gene was characterized using bioinformatics and Bayesian Inference. RESULTS: The 40 patients analyzed had a mean age of 45.2 years and 70% presented some type of symptom, with a predominance of pain and sensitivity, dysautonomia, and motor disorders. All patients presented the aA (Transcontinental Cosmopolitan) genotype, with an extremely low mutation rate, which is characteristic of the codifying region (aA - 1.83 × 10-4 mutations per site per year). The gp46 gene had a nucleotide diversity of between 0.00% and 2.0%. Amino acid mutations were present in 66.6% of the samples of individuals with signs/symptoms or diseases associated with HTLV-1 (p = 0.0091). Of the three most frequent mutations, the previously undescribed N93D mutant was invariably associated with symptomatic cases. CONCLUSIONS: The aA HTLV-1 subtype is predominant in the metropolitan region of Belém and presented a high degree of genetic stability in the codifying region. The rare N93D amino acid mutation may be associated with the clinical manifestations of this viral infection. IMPORTANCE: Little is known of the phylogeny of HTLV-1 in the endemic region of Brazilian Amazonia, and few complete gene sequences are available for the gp46 glycoprotein from the local population. The nucleotide sequences of the viral gp46 gene recorded in the present study confirmed the genetic stability of the region, and pointed to a homogeneous viral group, with local geographic characteristics. Further research will be necessary to more fully understand the molecular diversity of this protein, given the potential of this codifying region as a model for an effective HTLV-1 vaccine. The identification of a rare mutation (N93D), present only in symptomatic patients, should also be investigated further as a potential clinical marker. TRIAL REGISTRATION: ISRCTN 12345678, registered 28 September 2014.

Low genetic diversity of the Human T-cell Lymphotropic Virus (HTLV-1) in an endemic area of the Brazilian Amazon basin.

The Human T-cell Lymphotropic Virus (HTLV-1) is a Deltaretrovírus that was first isolated in the 1970s, and associated with Adult T-cell Leucemia-Lymphoma (ATLL), and subsequently to Tropical Spastic Paraparesis-Myelopathy (TSP/HAM). The genetic diversity of the virus varies among geographic regions, although its mutation rate is very low (approximately 1% per thousand years) in comparison with other viruses. The present study determined the genetic diversity of HTLV-1 in the metropolitan region of Belém, in northern Brazil. Blood samples were obtained from patients at the UFPA Tropical Medicine Nucleus between January 2010 and December 2013. The DNA was extracted and the PX region of the HTLV was amplified using nested PCR. The positive samples were then digested using the Taq1 enzyme for the identification and differentiation of the HTLV-1 and HTLV-2. The 5'LTR region of the positive HTLV-1 samples were amplified by nested PCR, and then sequenced genetically. The phylogenetic analysis of the samples was based on the maximum likelihood method and the evolutionary profile was analyzed by the Bayesian approach. Overall, 78 samples tested positive for HTLV-1, and 44 were analyzed here. The aA (cosmopolitan-transcontinental) subtype was recorded in all the samples. The following evolutionary rates were recorded for the different subtypes-a: 2.10-3, b: 2.69. 10-2, c: 6.23. 10-2, d: 3.08. 10-2, e: 6. 10-2, f: 1.78. 10-3, g: 2.2. 10-2 mutations per site per year. The positive HTLV-1 samples tested in the present study were characterized by their low genetic diversity and high degree of stability.

Health-related quality of life of children and adolescents with osteogenesis imperfecta: a cross-sectional study using PedsQL™.

BACKGROUND: Osteogenesis imperfecta (OI) is a disorder of bone formation leading to low mineral density and fractures. Children and adolescents with OI require periodic medical follow up, corrective surgery, drug therapy and physical therapy, as well as specific daily care practices. In addition, they have an increased incidence of fractures, which require immobilization and cause severe discomfort and short-term disability. This study evaluated the health-related quality of life of children and adolescents with OI in two reference centers for OI treatment in southern Brazil. METHODS: In this prospective cross-sectional study, the Pediatric Quality of Life Inventory (PedsQLTM) was applied in two university-affiliated reference centers for OI treatment in southern Brazil. Children and adolescents aged ≥ 5 years with clinical diagnoses of OI were included. Clinical data and socioeconomic status was evaluated. RESULTS: The sample consisted of 52 children and adolescents with OI (aged 5-17 years); 26 (50%) participants with type I OI, 13 (25%) type IV, 12 (23.1 %) type III, and 1 (1.9%) type V OI. Physical and social functioning domains differed significantly according to clinical presentation of OI with lowest scores in the severe type (OI type III). Pain seems to be the variable that is most associated with impact on the PedsQL domains. CONCLUSIONS: Overall, this study revealed differences in physical functioning and social functioning in relation to OI clinical presentation. These results reinforcing the importance of the clinical management of these patients with the aim of functional improvement and importance of pain control.