CHEK2 germline variants identified in familial nonmedullary thyroid cancer lead to impaired protein structure and function.

Approximately 5 to 15% of nonmedullary thyroid cancers (NMTC) present in a familial form (familial nonmedullary thyroid cancers [FNMTC]). The genetic basis of FNMTC remains largely unknown, representing a limitation for diagnostic and clinical management. Recently, germline mutations in DNA repair-related genes have been described in cases with thyroid cancer (TC), suggesting a role in FNMTC etiology. Here, two FNMTC families were studied, each with two members affected with TC. Ninety-four hereditary cancer predisposition genes were analyzed through next-generation sequencing, revealing two germline CHEK2 missense variants (c.962A > C, p.E321A and c.470T > C, p.I157T), which segregated with TC in each FNMTC family. p.E321A, located in the CHK2 protein kinase domain, is a rare variant, previously unreported in the literature. Conversely, p.I157T, located in CHK2 forkhead-associated domain, has been extensively described, having conflicting interpretations of pathogenicity. CHK2 proteins (WT and variants) were characterized using biophysical methods, molecular dynamics simulations, and immunohistochemistry. Overall, biophysical characterization of these CHK2 variants showed that they have compromised structural and conformational stability and impaired kinase activity, compared to the WT protein. CHK2 appears to aggregate into amyloid-like fibrils in vitro, which opens future perspectives toward positioning CHK2 in cancer pathophysiology. CHK2 variants exhibited higher propensity for this conformational change, also displaying higher expression in thyroid tumors. The present findings support the utility of complementary biophysical and in silico approaches toward understanding the impact of genetic variants in protein structure and function, improving the current knowledge on CHEK2 variants' role in FNMTC genetic basis, with prospective clinical translation.

Identification of Germline FOXE1 and Somatic MAPK Pathway Gene Alterations in Patients with Malignant Struma Ovarii, Cleft Palate and Thyroid Cancer.

Germline variants in the FOXE1 transcription factor have been associated with thyroid ectopy, cleft palate (CP) and thyroid cancer (TC). Here, we aimed to clarify the role of FOXE1 in Portuguese families (F1 and F2) with members diagnosed with malignant struma ovarii (MSO), an ovarian teratoma with ectopic malignant thyroid tissue, papillary TC (PTC) and CP. Two rare germline heterozygous variants in the FOXE1 promoter were identified: F1) c.-522G>C, in the proband (MSO) and her mother (asymptomatic); F2) c.9C>T, in the proband (PTC), her sister and her mother (CP). Functional studies using rat normal thyroid (PCCL3) and human PTC (TPC-1) cells revealed that c.9C>T decreased FOXE1 promoter transcriptional activity in both cell models, while c.-522G>C led to opposing activities in the two models, when compared to the wild type. Immunohistochemistry and RT-qPCR analyses of patients' thyroid tumours revealed lower FOXE1 expression compared to adjacent normal and hyperplastic thyroid tissues. The patient with MSO also harboured a novel germline AXIN1 variant, presenting a loss of heterozygosity in its benign and malignant teratoma tissues and observable ß-catenin cytoplasmic accumulation. The sequencing of the F1 (MSO) and F2 (PTC) probands' tumours unveiled somatic BRAF and HRAS variants, respectively. Germline FOXE1 and AXIN1 variants might have a role in thyroid ectopy and cleft palate, which, together with MAPK pathway activation, may contribute to tumours' malignant transformation.

Clinical and molecular characterisation of metastatic papillary thyroid cancer according to radioiodine therapy outcomes.

PURPOSE: Radioiodine (RAI) therapy remains the gold-standard approach for distant metastatic differentiated thyroid cancer (TC). The main objective of our work was to identify the clinical and molecular markers that may help to predict RAI avidity and RAI therapy response of metastatic lesions in a cohort of papillary thyroid cancer (PTC) patients. METHODS: We performed a retrospective analysis of 122 PTC patients submitted to RAI therapy due to distant metastatic disease. We also analysed, through next-generation sequencing, a custom panel of 78 genes and rearrangements, in a smaller cohort of 31 metastatic PTC, with complete follow-up, available RAI therapy data, and existing tumour sample at our centre. RESULTS: The most frequent outcome after RAI therapy was progression of disease in 59.0% of cases (n = 71), with median estimate progression-free survival of 30 months. RAI avidity was associated with PTC subtype, age and stimulated thyroglobulin at first RAI therapy for metastatic disease. The most frequently altered genes in the cohort of 31 PTC patients' primary tumours were RAS isoforms (54.8%) and TERT promoter (TERTp) (51.6%). The presence of BRAF p.V600E or RET/PTC alterations was associated with lower avidity (p = 0.012). TERTp mutations were not associated with avidity (p = 1.000) but portended a tendency for a higher rate of progression (p = 0.063); similar results were obtained when RAS and TERTp mutations coexisted (p = 1.000 and p = 0.073, respectively). CONCLUSIONS: Early identification of molecular markers in primary tumours may help to predict RAI therapy avidity, the response of metastatic lesions and to select the patients that may benefit the most from other systemic therapies.

SPRY4 as a Potential Mediator of the Anti-Tumoral Role of Macrophages in Anaplastic Thyroid Cancer Cells.

Anaplastic thyroid carcinoma (ATC) is the most lethal subtype of thyroid cancer, with high invasive and metastatic potential, not responding to conventional treatments. Its aggressiveness may be influenced by macrophages, which are abundant cells in the tumor microenvironment. To investigate the role of macrophages in ATC aggressiveness, indirect co-cultures were established between ATC cell lines and THP-1-derived macrophages. Macrophages significantly increased both the migration and invasion of T235 cells (p < 0.01; p < 0.01), contrasting with a decrease in C3948 (p < 0.001; p < 0.05), with mild effects in T238 migration (p < 0.01) and C643 invasion (p < 0.05). Flow cytometry showed upregulation of CD80 (pro-inflammatory, anti-tumoral) and downregulation of CD163 (anti-inflammatory, pro-tumoral) in macrophages from co-culture with T235 (p < 0.05) and C3948 (p < 0.05), respectively. Accordingly, we found an upregulation of secreted pro-inflammatory mediators (e.g., GM-CSF, IL-1α; p < 0.05) in C3948-macrophage co-cultures. Proteomic analysis showed the upregulation of SPRY4, an inhibitor of the MAPK pathway, in C3948 cells from co-culture. SPRY4 silencing promoted cancer cell invasion, reverting the reduced invasion of C3948 caused by macrophages. Our findings support that macrophages play a role in ATC cell aggressiveness. SPRY4 is a possible modulator of macrophage-ATC cell communication, with a tumor suppressor role relevant for therapeutic purposes.

Target therapy for BRAF mutated anaplastic thyroid cancer: a clinical and molecular study.

OBJECTIVES: Anaplastic thyroid carcinoma (ATC) has a poor survival. The combination of Dabrafenib plus Trametinib (DT) had a significant impact in survival of BRAF p.V600E patients. However, durable responses may be compromised by resistance. We aim to present our experience with DT in BRAF positive ATC patients and compare the outcomes with usual therapy, and to study tumor molecular alterations in the DT group. METHODS: Patients treated between May 2018 and April 2022 in a tertiary referral center, assessed for BRAF status were included. Patients were divided in three groups: BRAF p.V600E treated with DT, BRAF wild type (WT) under multimodal therapy (MT), and BRAF WT under compassionate care (CC). Response was assessed monthly in the first 6 months and every 3 months afterwards, by RECIST 1.1. Overall survival (OS) and progression-free survival (PFS) were estimated with the Kaplan-Meier method and compared with the log-rank test. RESULTS: Twenty-seven ATC patients were included (DT = 9, MT = 8, and CC = 10). Median OS was 475 days for DT, 156 days for MT, and 39 days for CC (P < .001). At 12 months, only patients in the DT group were alive (71%). Median PFS was 270 days, in the DT group, compared with less than 32 days in BRAF WT (P < .001). No severe adverse events were reported. Molecular profiling showed that in one of the four clinical progressions, a pathogenic NRAS mutation was found. CONCLUSIONS: Our results show a significant real-world efficacy of Dabrafenib plus Trametinib in both survival and recurrence compared with standard treatment, with a good safety profile.

Systematics and biogeography of the Atlantic Forest endemic genus Juliomys (Rodentia: Cricetidae): a test of diversification hypothesis using mitochondrial data

The Atlantic Forest harbors a large species richness and high levels of endemism, but the processes that shaped its biodiversity are poorly studied, especially for mammals. Among them are the endemic mice Juliomys, which comprise forest dwellers distributed in southeastern and southern Brazil, northeastern Argentina, and eastern Paraguay. In this study, we investigate the phylogenetic relationships among species and perform phylogeographic analyses to evaluate the population structure and demographic scenarios through mitochondrial gene cytochrome b sequences. We investigate three hypotheses of diversification (forest refuges, montane isolate, and geomorphological events) to understand the evolution of the Juliomys species. Phylogenetic analyses recovered five clades/lineages, four of which are congruent with species currently recognized. The fifth lineage expands the range of the genus 659 km to the north and may represent a new species. The observed demographic and geographic structure of genetic diversity does not match the forest refuge hypothesis as mechanism to explain the diversification in Juliomys. Our results recovered J. rimofrons and J. ximenezi as sister species, supporting predictions of montane isolate hypothesis. We also detected a shallow genetic structure in J. pictipes and J. ossitenuis. Both phylogeographic breaks were congruent with limits of the São Paulo Basin, an area that has undergone Neogene reactivations of tectonic faults. It is suggested that geomorphological events led to a deformed landscape that influenced the dynamics of sedimentary basins and promoted an incipient population structure in J. pictipes and J. ossitenuis. Our findings demonstrate that the divergences whithin Juliomys species occurred during the Quaternary, too recently to have produced strong geographic structure.

Systematics and biogeography of the atlantic forest endemic genus Juliomys (Rodentia: Cricetidae): A test of diversification hypothesis using mitochondrial data

The Atlantic Forest harbors a large species richness and high levels of endemism, but the processes that shaped its biodiversity are poorly studied, especially for mammals. Among them are the endemic mice Juliomys, which comprise forest dwellers distributed in southeastern and southern Brazil, northeastern Argentina, and eastern Paraguay. In this study, we investigate the phylogenetic relationships among species and perform phylogeographic analyses to evaluate the population structure and demographic scenarios through mitochondrial gene cytochrome b sequences. We investigate three hypotheses of diversification (forest refuges, montane isolate, and geomorphological events) to understand the evolution of the Juliomys species. Phylogenetic analyses recovered five clades/lineages, four of which are congruent with species currently recognized. The fifth lineage expands the range of the genus 659 km to the north and may represent a new species. The observed demographic and geographic structure of genetic diversity does not match the forest refuge hypothesis as mechanism to explain the diversification in Juliomys. Our results recovered J. rimofrons and J. ximenezi as sister species, supporting predictions of montane isolate hypothesis. We also detected a shallow genetic structure in J. pictipes and J. ossitenuis. Both phylogeographic breaks were congruent with limits of the São Paulo Basin, an area that has undergone Neogene reactivations of tectonic faults. It is suggested that geomorphological events led to a deformed landscape that influenced the dynamics of sedimentary basins and promoted an incipient population structure in J. pictipes and J. ossitenuis. Our findings demonstrate that the divergences whithin Juliomys species occurred during the Quaternary, too recently to have produced strong geographic structure.

Kawasaki Disease Associated with Covid-19 in a Pediatric Patient.

Kawasaki disease (KD) is a multisystemic vasculitis of small and medium-size vessels that mainly affects children under five years of age. The etiology of KD is still uncertain; however, evidence suggests that infectious agents and genetic susceptibility act as a trigger for its development, which could explain the increase in cases of children with COVID-19 who developed a classic or incomplete form of KD. The aim of this report is to discuss a case of a five-year-old boy diagnosed with incomplete KD associated with COVID-19.

Burden, family functioning, and psychological health of older caregivers of older adults: a path analysis / Sobrecarga, funcionalidade familiar e saúde psicológica de idosos cuidadores de idosos: uma path analysis

Objectives: To evaluate an explanatory model of direct and indirect associations regarding the psychological health of older caregivers of functionally dependent older adults. Methods: This is a cross-sectional study performed with older caregivers recruited in contexts of outpatient and home care. We collected information on sociodemographic characteristics, duration of caregiving, physical and cognitive function indicators of the older care recipients, perceived burden, family functioning, and psychological health measures (psychological need satisfaction and depressive symptoms). Results: We evaluated 133 caregivers (76% female, 69.5 ± 6.98 years). Variables that were significantly correlated with psychological health were selected to form an association model to be tested by structural equation modeling via path analysis. Depressive symptom variability was best explained by this model. Caregiver burden remained in the model as a mediator of indirect associations between physical function for instrumental activities of daily living and indicators of family functioning and psychological health. Three associative paths between caregiver burden and depressive symptoms were found ­ one of them was direct and the other two were mediated whether by family functioning or by the level of psychological need satisfaction. Conclusion: Depressive symptoms were the psychological health indicator best explained by the model involving instrumental functional demands that generate burden. Clinical consequences suggested by the model indicate interventions aimed at family functioning and opportunities of psychological need satisfaction as strategies for promoting caregivers' psychological health.

Objetivos: Avaliar um modelo de associações diretas e indiretas explicativo de saúde psicológica de idosos cuidadores de outros idosos funcionalmente dependentes. Metodologia: Estudo transversal realizado com idosos cuidadores recrutados em contexto ambulatorial e de atenção domiciliar. Foram levantadas informações sociodemográficas, tempo de exercício do cuidado, indicadores funcionais físicos e cognitivos dos idosos que recebem os cuidados, percepção de sobrecarga, funcionalidade familiar e medidas de saúde psicológica (satisfação de necessidades psicológicas e sintomatologia depressiva). Resultados: Foram avaliados 133 cuidadores (76% feminino, 69,5 ± 6,98 anos). As variáveis correlacionadas significativamente à saúde psicológica foram escolhidas para a composição do modelo de associações testado por análise de equações estruturais via path analysis. A variabilidade em sintomatologia depressiva foi melhor explicada pelo modelo. Sobrecarga permaneceu no modelo como mediadora das associações indiretas entre funcionalidade física para atividades instrumentais da vida diária e os indicadores de funcionalidade familiar e de saúde psicológica. Três caminhos associativos entre sobrecarga e sintomatologia depressiva foram encontrados ­ um direto e dois mediados, ora pela funcionalidade familiar, ora pelo nível de satisfação de necessidades psicológicas. Conclusão: Sintomatologia depressiva foi o indicador de saúde psicológica mais bem explicado pelo modelo que envolve a presença de demandas funcionais instrumentais geradoras de sobrecarga. Desdobramentos clínicos sugeridos pelo modelo apontam para intervenções com alvo em funcionalidade familiar e de oportunidades de satisfação de necessidades psicológicas como estratégias para promover a saúde psicológica do cuidador.

Identification of SPRY4 as a Novel Candidate Susceptibility Gene for Familial Nonmedullary Thyroid Cancer.

Background: The molecular basis of familial nonmedullary thyroid cancer (FNMTC) is still poorly understood, representing a limitation for molecular diagnosis and clinical management. In this study, we aimed to identify new susceptibility genes for FNMTC through whole-exome sequencing (WES) analysis of leukocyte DNA of patients from a highly informative FNMTC family. Methods: We selected six affected family members to conduct WES analysis. Bioinformatic analyses were undertaken to filter and select the genetic variants shared by the affected members, which were subsequently validated by Sanger sequencing. To select the most likely pathogenic variants, several studies were performed, including family segregation analysis, in silico impact characterization, and gene expression (messenger RNA and protein) depiction in databases. For the most promising variant identified, we performed in vitro studies to validate its pathogenicity. Results: Several potentially pathogenic variants were identified in different candidate genes. After filtering with appropriate criteria, the variant c.701C>T, p.Thr234Met in the SPRY4 gene was prioritized for in vitro functional characterization. This SPRY4 variant led to an increase in cell viability and colony formation, indicating that it confers a proliferative advantage and potentiates clonogenic capacity. Phosphokinase array and Western blot analyses suggested that the effects of the SPRY4 variant were mediated through the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway, which was further supported by a higher responsiveness of thyroid cancer cells with the SPRY4 variant to a MEK inhibitor. Conclusions: WES analysis in one family identified SPRY4 as a likely novel candidate susceptibility gene for FNMTC, allowing a better understanding of the cellular and molecular mechanisms underlying thyroid cancer development.

A pathogenic variant in CHEK2 shows a founder effect in Portuguese Roma patients with thyroid cancer.

PURPOSE: Germline mutations in DNA repair-related genes have been recently reported in cases with familial non-medullary thyroid carcinoma (FNMTC). A Portuguese family from the Roma ethnic group with four members affected with papillary thyroid carcinoma (PTC), and three members with multinodular goiter (MNG) was identified. The aim of this study was to investigate the involvement of DNA repair-related genes in the etiology of FNMTC in this family and in the Roma ethnic group. METHODS: Ninety-four hereditary cancer predisposition genes were analyzed through next-generation sequencing. Sanger sequencing was used for variant confirmation and screening. Twelve polymorphic markers were genotyped for haplotype analysis in the CHEK2 locus. RESULTS: A germline pathogenic frameshift variant in the CHEK2 gene [c.596dupA, p.(Tyr199Ter)] was detected in homozygosity in the proband (PTC) and in his brother (MNG), being heterozygous in his mother (PTC), two sisters (PTC), and one nephew (MNG). This variant was absent in 100 general population controls. The screening of the CHEK2 variant was extended to other Roma individuals, being detected in 2/33 Roma patients with thyroid cancer, and in 1/15 Roma controls. Haplotype segregation analysis identified a common ancestral core haplotype (Hcac), covering 10 Mb in the CHEK2 locus, shared by affected CHEK2 variant carriers. Analysis of 62 individuals CHEK2 wild-type indicated that none presented the Hcac haplotype. The estimated age for this variant suggested that it was transmitted by a relatively recent common ancestor. CONCLUSIONS: We identified a founder CHEK2 pathogenic variant, which is likely to underlie thyroid cancer and other cancer manifestations in the Roma population.

Clinical and post mortem examination of white worm lizards (Amphisbaena alba) in the State of Paraíba, Northeastern Brazil: morphological, pathological and radiographic findings of a secretive species / Exame clínico e post mortem de anfisbenas (Amphisbaena alba) no Estado da Paraíba, Nordeste do Brasil: achados morfológicos, patológicos e radiográficos de uma espécie reservada

Amphisbenians are limbless reptiles that belong to the order Squamata. Due to their fossorial and secrevie habits, little is known about their morphology, ecology and pathological conditions that may affect them. In this manuscript, we present a brief guide for identification of normal structures as well as traumatic injuries on radiography and necropsy of Amphisbaena alba. From April to September 2019, three cases of A. alba with suspected trauma were referred to the Veterinary Hospital of the Federal University of Paraíba (UFPB). In the clinical evaluation, traumatic injuries were observed, and support therapy was instituted, but they did not resist and died shortly after. Bone fractures and organ ruptures, in addition to specific structures of this species were identified on radiography. A systematic necropsy was performed of all amphisbaenians in order to evaluate external and internal structures, not only to identify lesions but also to investigate the morphological aspects of amphisbenids. Macroscopically, multiple organ fractures and ruptures observed in radiographs were confirmed, in addition to the presence of the cestodes Semenoviella amphisbaenae in the large intestine. Histologically, it was possible to identify normal characteristics and microscopic lesions in the tissues. This is the first study to incorporate morphological, clinical, and pathological aspects of A. alba. This manuscript brings essential information for wildlife veterinarians and pathologists who may have to treat or perform a necropsy on these unique reptiles.(AU)

Anfisbenas são répteis desprovidos de membros que pertencem a ordem Squamata. Devido a seus hábitos fossoriais e reclusos, pouco se conhece sobre sua morfologia, ecologia e condições patológicas que possam afetá-las. Neste artigo, um guia sucinto para a necropsia, identificação de tecidos e órgãos e lesões encontradas em Amphisbaena alba, é apresentado. Durante março e dezembro de 2019, três casos de A. alba com suspeita de trauma foram encaminhados ao Hospital Veterinário da Universidade Federal da Paraíba. Na avaliação clínica foram observadas lesões traumáticas e instituído um suporte terapêutico, porém não resistiram e morreram em seguida. Foram realizadas projeções radiográficas onde identificaram fraturas e ruptura de órgãos, além de identificar estruturas específicas dessa espécie. Uma avaliação completa de estruturas externas e internas foi conduzida para investigar aspectos morfológicos dos anfisbenídeos. Macroscopicamente foram confirmadas múltiplas fraturas e rupturas de órgãos observadas anteriormente nas radiografias, além disso evidenciou no intestino grosso presença de parasitos anoplocéfalos denominados Semenoviella amphisbaenae. Na histologia foi possível identificar características normais e lesões microscópicas nos tecidos. Esse é o primeiro estudo a incorporar aspectos morfológicos, clínicos e patológicos de A. alba. Esse manuscrito traz informações essenciais para clínicos e patologistas de animais selvagens que podem se deparar com casos clínicos ou de necropsia de Amphisbaena alba.(AU)

Clinical and post mortem examination of white worm lizards (Amphisbaena alba) in the State of Paraíba, Northeastern Brazil: morphological, pathological and radiographic findings of a secretive species

ABSTRACT: Amphisbenians are limbless reptiles that belong to the order Squamata. Due to their fossorial and secrevie habits, little is known about their morphology, ecology and pathological conditions that may affect them. In this manuscript, we present a brief guide for identification of normal structures as well as traumatic injuries on radiography and necropsy of Amphisbaena alba. From April to September 2019, three cases of A. alba with suspected trauma were referred to the Veterinary Hospital of the Federal University of Paraíba (UFPB). In the clinical evaluation, traumatic injuries were observed, and support therapy was instituted, but they did not resist and died shortly after. Bone fractures and organ ruptures, in addition to specific structures of this species were identified on radiography. A systematic necropsy was performed of all amphisbaenians in order to evaluate external and internal structures, not only to identify lesions but also to investigate the morphological aspects of amphisbenids. Macroscopically, multiple organ fractures and ruptures observed in radiographs were confirmed, in addition to the presence of the cestodes Semenoviella amphisbaenae in the large intestine. Histologically, it was possible to identify normal characteristics and microscopic lesions in the tissues. This is the first study to incorporate morphological, clinical, and pathological aspects of A. alba. This manuscript brings essential information for wildlife veterinarians and pathologists who may have to treat or perform a necropsy on these unique reptiles.

RESUMO: Anfisbenas são répteis desprovidos de membros que pertencem a ordem Squamata. Devido a seus hábitos fossoriais e reclusos, pouco se conhece sobre sua morfologia, ecologia e condições patológicas que possam afetá-las. Neste artigo, um guia sucinto para a necropsia, identificação de tecidos e órgãos e lesões encontradas em Amphisbaena alba, é apresentado. Durante março e dezembro de 2019, três casos de A. alba com suspeita de trauma foram encaminhados ao Hospital Veterinário da Universidade Federal da Paraíba. Na avaliação clínica foram observadas lesões traumáticas e instituído um suporte terapêutico, porém não resistiram e morreram em seguida. Foram realizadas projeções radiográficas onde identificaram fraturas e ruptura de órgãos, além de identificar estruturas específicas dessa espécie. Uma avaliação completa de estruturas externas e internas foi conduzida para investigar aspectos morfológicos dos anfisbenídeos. Macroscopicamente foram confirmadas múltiplas fraturas e rupturas de órgãos observadas anteriormente nas radiografias, além disso evidenciou no intestino grosso presença de parasitos anoplocéfalos denominados Semenoviella amphisbaenae. Na histologia foi possível identificar características normais e lesões microscópicas nos tecidos. Esse é o primeiro estudo a incorporar aspectos morfológicos, clínicos e patológicos de A. alba. Esse manuscrito traz informações essenciais para clínicos e patologistas de animais selvagens que podem se deparar com casos clínicos ou de necropsia de Amphisbaena alba.

Morphological variation in the genus Juliomys (Rodentia: Cricetidae: Sigmodontinae) and taxonomic status of Juliomys anoblepas (Winge 1887) from the Quaternary of Southeast Brazil.

The region of Lagoa Santa, Minas Gerais, Brazil, is one of the most important karstic areas of the Brazilian Quaternary due to the faunistic diversity of living and extinct forms. Among them, some taxa remain poorly studied, as is the case of Calomys anoblepas Winge 1887. Despite the recent allocation of the taxon within Juliomys, its description and morphological analysis are condensed, based on comparative few specimens and on few informative characters. In this study, we investigate characters proposed to distinguish species of Juliomys, and reevaluate the taxonomic status of the fossil Juliomys anoblepas. We analyzed 80 cranio-dental morphological characters in 233 specimens represented by the four species currently recognized: J. pictipes (Osgood 1933), J. rimofrons Oliveira Bonvicino 2002, J. ossitenuis Costa, Pavan, Leite Fagundes 2007, and J. ximenezi Christoff, Vieira, Oliveira, Gonçalves, Valiati Tomasi 2016. We also performed principal component analysis on eight craniodental measurements available for the J. anoblepas hypodigm. The review of morphological systems and the evaluation of the characters used in the literature revealed that there are no diagnostic characters in the anterior portion of the skull and in the molar series of Juliomys, being difficult to differentiate the fossil from the other living species. Only six qualitative characters were variable and applicable to the hypodigm of J. anoblepas. Characters are polymorphic, invariable, or the fossil is not sufficiently complete to determinate its states. The taxon could not be morphometrically differentiated from J. pictipes and J. ossitenuis. Based on the results presented herein, we consider J. anoblepas as a nomen dubium and restrict its name to the taxon's hypodigm.

Safety, Tumor Reduction, and Clinical Impact of Zika Virus Injection in Dogs with Advanced-Stage Brain Tumors.

Malignant brain tumors are among the most aggressive cancers with poor prognosis and no effective treatment. Recently, we reported the oncolytic potential of Zika virus infecting and destroying the human central nervous system (CNS) tumors in vitro and in immunodeficient mice model. However, translating this approach to humans requires pre-clinical trials in another immunocompetent animal model. Here, we analyzed the safety of Brazilian Zika virus (ZIKVBR) intrathecal injections in three dogs bearing spontaneous CNS tumors aiming an anti-tumoral therapy. We further assessed some aspects of the innate immune and inflammatory response that triggers the anti-tumoral response observed during the ZIKVBR administration in vivo and in vitro. For the first time, we showed that there were no negative clinical side effects following ZIKVBR CNS injections in dogs, confirming the safety of the procedure. Furthermore, the intrathecal ZIKVBR injections reduced tumor size in immunocompetent dogs bearing spontaneous intracranial tumors, improved their neurological clinical symptoms significantly, and extended their survival by inducing the destruction specifically of tumor cells, sparing normal neurons, and activating an immune response. These results open new perspectives for upcoming virotherapy using ZIKV to destroy and induce an anti-tumoral immune response in CNS tumors for which there are currently no effective treatments.

Atenção nutricional e práticas alimentares na perspectiva de gestantes com excesso de peso / Nutritional attention and food practices in the perspective of overweight pregnant women

Objetivo: Este estudo tem como objetivo investigar a atenção nutricional e as práticas alimentares, na perspectiva de gestantes com excesso de peso assistidas na Atenção Básica de Macaé, Rio de Janeiro. Métodos: Trata-se de pesquisa de abordagem qualitativa, fundamentada no paradigma interpretativista. Realizaram-se entrevistas em profundidade, cujos dados foram submetidos à análise de conteúdo temática adaptada de Bardin. Foram entrevistadas 12 gestantes com excesso de peso, maiores de 20 anos, residentes em Macaé-RJ. Resultados: O estudo verificou o estabelecimento de relações verticalizadas entre o profissional de saúde-usuário, descontinuidade do atendimento nutricional e a não adesão às orientações dietéticas por algumas participantes. Houve maior aceitação do ganho de peso gestacional excessivo e adoção de práticas alimentares ambíguas, permeadas tanto pelo consumo exagerado de alimentos quanto pela retirada de alimentos considerados "ruins e/ou besteiras". O discurso verticalizado do profissional de saúde, frequentemente autoritário, e a anulação da mulher enquanto sujeito ativo na construção do cuidado se manifestaram, de forma implícita, nas narrativas. Conclusão: A naturalização do excesso de peso gestacional, concepções construídas no imaginário social como o "comer por dois" e "desejos" durante a gestação, o conjunto das práticas alimentares nesse período e o suporte familiar constituem elementos relevantes a serem considerados pelas equipes de saúde para a organização da atenção nutricional. (AU)

Objective: The aim of this study is to investigate nutritional care and dietary practices, from the perspective of overweight pregnant women receiving primary health care in the town of Macaé, Rio de Janeiro state. Methods: This is a qualitative research study, based on the interpretive paradigm. In-depth interviews were conducted, and the data underwent thematic content analysis, adapted from Bardin. Twelve overweight pregnant women, over 20 years old, living in Macaé-RJ, were interviewed. Results: The study found the establishment of vertical relationships between the health workerpatient, discontinued nutritional care and non-adherence to dietary guidelines by some participants. There was greater acceptance of excessive gestational weight gain and the adoption of ambiguous eating practices, permeated both by the excessive intake of food and by the withdrawal of foods considered to be "bad and/or junk". The vertical discourse of health workers, which is often authoritarian, and the nullification of women as an active subject in the construction of care, were implicitly reported in the narratives. Conclusion: Naturalization of gestational excess weight, conceptions built in the social imaginary such as "eating for two" and "cravings" during pregnancy, the set of eating practices in this period and family support are relevant elements to be considered by health teams for the organization of nutritional care. (AU)

Evolução do excesso de peso em gestantes usuárias da Atenção Primária à Saúde do município de Macaé-RJ entre 2010-2018 / Evolution of overweight pregnant women in the Primary Health Care in the city of Macaé-RJ between 2010-2018

Introdução: O estudo visa investigar a taxa de prevalência e a tendência temporal do excesso de peso em gestantes usuárias da Atenção Primária à Saúde de Macaé, entre 2010 e 2018. Método: Realizou-se estudo quantitativo, de série temporal, com a base de dados do Sistema de Vigilância Alimentar e Nutricional (Sisvan WEB), de acesso público, referente ao diagnóstico nutricional de gestantes ≥20 anos usuárias da Atenção Primária à Saúde do município de Macaé, no período de 2010 a 2018. Analisaram-se 4.279 registros de gestantes no período de nove anos. As variáveis estudadas foram: ano de coleta e registro dos dados; baixo peso; peso adequado; sobrepeso; obesidade; excesso de peso (sobrepeso e obesidade). A análise de tendência do excesso de peso foi realizada por meio de modelos de regressão linear simples, considerando-se o nível de significância de 5%. Resultados: As prevalências do excesso de peso em gestantes foram elevadas e ascendentes: 2010 (40,2%), 2011 (37,5%), 2012 (46,8%), 2013 (48,8%), 2014 (50,9%), 2015 (51,8%), 2016 (54,4%), 2017 (57,6%) e 2018 (55,8%). Em todas as análises, detectou-se a tendência significativa e crescente, podendo-se destacar o desfecho do excesso de peso, que apresentou velocidade de incremento anual (ß1=2,35; p-valor<0,001) superior aos demais desfechos analisados em separado. Conclusão: Conclui-se que é elevada e crescente a prevalência do excesso de peso em gestantes usuárias da Atenção Primária à Saúde do município de Macaé no período estudado. (AU)

Introduction The study aims to investigate the prevalence rate and the time trend of excess weight in pregnant women receiving Primary Health Care in Macaé, between 2010 and 2018. Method: A quantitative, time series study was conducted with the open access database of the Food and Nutritional Surveillance System (Sisvan WEB), relative to the nutritional diagnosis of pregnant women> 20 years old, receiving Primary Health Care in the city of Macaé, between 2010 and 2018. Analyses were made of 4,279 records of pregnant women over a nine-year period. The study variables were: year of data collection and registration; low weight; adequate weight; overweight; obesity; excess weight (overweight and obesity). Trend analysis of excess weight was performed using simple linear regression models, considering the significance level of 5%. Results: Prevalence of overweight in pregnant women was high and rising: 2010 (40.2%), 2011 (37.5%), 2012 (46.8%), 2013 (48.8%), 2014 (50.9%), 2015 (51.8%), 2016 (54.4%), 2017 (57.6%) and 2018 (55.8%). All analyses showed a significant and growing trend, especially for the excess weight outcome, whose annual increase speed (£ 1 = 2.35; p-value <0.001) was higher than the other outcomes, when analyzed separately. Conclusion: Prevalence of excess weight in pregnant women receiving Primary Health Care in the city of Macaé was high and growing during the study period. (AU)

Microneedle Arrays of Polyhydroxyalkanoate by Laser-Based Micromolding Technique.

Although many delivery systems have been proposed to improve drug permeation through the skin, all of them suffer from several limitations. This drives a continuous search for innovative systems that can provide safe and effective transdermal drug delivery solutions. In line with this, microneedle (MN) arrays, a hybrid combination of hypodermic injections and transdermal patches, have been proposed. MNs consist of microscale needles that can pierce the skin by a simple, minimally invasive, and painless route, enabling the transport of drugs and macromolecules into the human body. This study reports, for the first time, the use of a biobased, biodegradable, and biocompatible polymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), P(3HB-co-3HV), for the fabrication of a biopolymer-based MN patch. Molds of poly(dimethylsiloxane) were prepared by direct laser writing technology, a low-cost and mask-less technology, and used to produce biodegradable P(3HB-co-3HV) MNs by a thermosetting process. The best results were obtained with a laser power of 30 W at 0.15 m/s with a spiral model as the pattern. The obtained MNs had a length of 0.69 mm and a diameter of 0.33 mm, ideal for painless penetration of the skin. Additionally, the produced MNs had good mechanical properties and the ability to be successfully impregnated with the fluorescent dye Rhodamine 6G. These features render P(3HB-co-3HV) a promising material for the development of MNs with improved functionality.