Characterizing lipid constituents of B. moojeni snake venom: a comparative approach for chemical and biological investigations.

Snake venoms are complex mixtures majorly composed of proteins with well-studied biological effects. However, the exploration of non-protein components, especially lipids, remains limited despite their potential for discovering bioactive molecules. This study compares three liquid-liquid lipid extraction methods for both chemical and biological analyses of Bothrops moojeni snake venom. The methods evaluated include the Bligh and Dyer method (methanol, chloroform, water), considered standard; the Acunha method, a modification of the Bligh and Dyer protocol; and the Matyash method (MTBE/methanol/water), featuring an organic phase less dense than the aqueous phase. Lipidomic analysis using liquid chromatography with high-resolution mass spectrometry (LC-HRMS) system revealed comparable values of lipid constituents' peak intensity across different extraction methods. Our results show that all methods effectively extracted a similar quantity of lipid species, yielding approximately 17-18 subclasses per method. However, the Matyash and Acunha methods exhibited notably higher proportions of biologically active lipids compared to the Bligh and Dyer method, particularly in extracting lipid species crucial for cellular structure and function, such as sphingomyelins and phosphatidylinositol-phosphate. In conclusion, when selecting a lipid extraction method, it is essential to consider the study's objectives. For a biological approach, it is crucial to evaluate not only the total quantity of extracted lipids but also their quality and biological activity. The Matyash and Acunha methods show promise in this regard, potentially offering a superior option for extracting biologically active lipids compared to the Bligh and Dyer method.

The Implications of Brain-Derived Neurotrophic Factor in the Biological Activities of Platelet-Rich Plasma.

Platelet-rich plasma (PRP) is a biological blood-derived therapeutic obtained from whole blood that contains higher levels of platelets. PRP has been primarily used to mitigate joint degeneration and chronic pain in osteoarthritis (OA). This clinical applicability is based mechanistically on the release of several proteins by platelets that can restore joint homeostasis. Platelets are the primary source of brain-derived neurotrophic factor (BDNF) outside the central nervous system. Interestingly, BDNF and PRP share key biological activities with clinical applicability for OA management, such as anti-inflammatory, anti-apoptotic, and antioxidant. However, the role of BDNF in PRP therapeutic activities is still unknown. Thus, this work aimed to investigate the implications of BDNF in therapeutic outcomes provided by PRP therapy in vitro and in-vivo, using the MIA-OA animal model in male Wistar rats. Initially, the PRP was characterized, obtaining a leukocyte-poor-platelet-rich plasma (LP-PRP). Our assays indicated that platelets activated by Calcium release BDNF, and suppression of M1 macrophage polarization induced by LP-PRP depends on BDNF full-length receptor, Tropomyosin Kinase-B (TrkB). OA animals were given LP-PRP intra-articular and showed functional recovery in gait, joint pain, inflammation, and tissue damage caused by MIA. Immunohistochemistry for activating transcriptional factor-3 (ATF-3) on L4/L5 dorsal root ganglia showed the LP-PRP decreased the nerve injury induced by MIA. All these LP-PRP therapeutic activities were reversed in the presence of TrkB receptor antagonist. Our results suggest that the therapeutic effects of LP-PRP in alleviating OA symptoms in rats depend on BDNF/TrkB activity.

Blocking Pannexin 1 Channels Alleviates Peripheral Inflammatory Pain but not Paclitaxel-Induced Neuropathy.

BACKGROUND: Pannexin1 (Panx1) is a membrane channel expressed in different cells of the nervous system and is involved in several pathological conditions, including pain and inflammation. At the central nervous system, the role of Panx1 is already well-established. However, in the periphery, there is a lack of information regarding the participation of Panx1 in neuronal sensitization. The dorsal root ganglion (DRG) is a critical structure for pain processing and modulation. For this reason, understanding the molecular mechanism in the DRG associated with neuronal hypersensitivity has become highly relevant to discovering new possibilities for pain treatment. Here, we aimed to investigate the role of Panx1 in acute nociception and peripheral inflammatory and neuropathic pain by using two different approaches. METHODS: Rats were treated with a selective Panx1 blocker peptide (10Panx) into L5-DRG, followed by ipsilateral intraplantar injection of carrageenan, formalin, or capsaicin. DRG neuronal cells were pre-treated with 10Panx and stimulated by capsaicin to evaluate calcium influx. Panx1 knockout mice (Panx1-KO) received carrageenan or capsaicin into the paw and paclitaxel intraperitoneally. The von Frey test was performed to measure the mechanical threshold of rats' and mice's paws before and after each treatment. RESULTS: Pharmacological blockade of Panx1 in the DRG of rats resulted in a dose-dependent decrease of mechanical allodynia triggered by carrageenan, and nociception decreased in the second phase of formalin. Nociceptive behavior response induced by capsaicin was significantly lower in rats treated with Panx1 blockade into DRG. Neuronal cells with Panx1 blockage showed lower intracellular calcium response than untreated cells after capsaicin administration. Accordingly, Panx1-KO mice showed a robust reduction in mechanical allodynia after carrageenan and a lower nociceptive response to capsaicin. A single dose of paclitaxel promoted acute mechanical pain in wildtype (WT) but not in Panx1-KO mice. Four doses of chemotherapy promoted chronic mechanical allodynia in both genotypes, although Panx1-KO mice had significant ablation in the first eight days. CONCLUSION: Our findings suggest that Panx1 is critical for developing peripheral inflammatory pain and acute nociception involving transient receptor potential vanilloid subtype 1 (TRPV1) but is not essential for neuropathic pain chronicity.

Prevalence and clinical implications of germline pathogenic variants in cancer predisposing genes in young patients across sarcoma subtypes.

BACKGROUND: Sarcomas are a rare and diverse group of cancers occurring mainly in young individuals for which an underlying germline genetic cause remains unclear in most cases. METHODS: Germline DNA from 177 children, adolescents and young adults with soft tissue or bone sarcomas was tested using multigene panels with 113 or 126 cancer predisposing genes (CPGs) to describe the prevalence of germline pathogenic/likely pathogenic variants (GPVs). Subsequent testing of a subset of tumours for loss of heterozygosity (LOH) evaluation was performed to investigate the clinical and molecular significance of these variants. RESULTS: GPVs were detected in 21.5% (38/177) of the patients (15.8% in children and 21.6% in adolescents and young adults), with dominant CPGs being altered in 15.2% overall. These variants were found in genes previously associated with the risk of developing sarcomas (TP53, RB1, NF1, EXT1/2) but also in genes where that risk is still emerging/limited (ERCC2, TSC2 and BRCA2) or unknown (PALB2, RAD50, FANCM and others). The detection rates of GPVs varied from 0% to 33% across sarcoma subtypes and GPV carriers were more likely to present more than one primary tumour than non-carriers (21.1%×6.5%; p=0.012). Loss of the wild-type allele was detected in 48% of tumours from GPV carriers, mostly in genes definitively associated with sarcoma risk. CONCLUSION: Our findings reveal that a high proportion of young patients with sarcomas presented a GPV in a CPG, underscoring the urgency of establishing appropriate genetic screening strategies for these individuals and their families.

Do the miniscrews of the prefabricated systems for the "appliance first" protocol of miniscrew-assisted rapid palatal expansion have lengths that meet the palatal thickness of patients?

INTRODUCTION: In the "appliance first" protocol of miniscrew-assisted rapid palatal expansion, the prefabricated shape of the expanders limits the potential locations for miniscrew placement. Considering the influence of palatal thickness on the selection of the optimal length of miniscrews, this study aimed to evaluate the thickness of both bone and mucosa of the palate of patients aged 6-65 years and suggest optimal lengths of miniscrews for this approach. METHODS: Two hundred and eighty-two cone-beam computed tomography of patients of both sexes were divided into 3 groups according to age. The thicknesses of bone and mucosa were measured in the anterior and posterior regions of the palate. RESULTS: Males showed a greater thickness of palatal bone than females. The thickness of both bone and mucosa was greater in the anterior region of the palate. The young patients showed greater bone thickness than adults and mature adults. The mature adults showed thinner bone thickness in the posterior region of the palate and greater mucosal thickness along the palate than young patients and adults. Development of miniscrews with longer thread lengths is necessary. Miniscrews with a 3-mm thread length would prevent excessive extravasation in the posterior region of the palate. CONCLUSIONS: The thickness of both bone and mucosa of the palate is variable and influenced by sex and age. Manufacturing additional miniscrews with different lengths of thread is suggested to achieve bicortical anchorage in patients undergoing the "appliance first" protocol of miniscrew-assisted rapid palatal expansion.

Brazilian Amazon indigenous territories under deforestation pressure.

Studies showed that Brazilian Amazon indigenous territories (ITs) are efficient models for preserving forests by reducing deforestation, fires, and related carbon emissions. Considering the importance of ITs for conserving socio-environmental and cultural diversity and the recent climb in the Brazilian Amazon deforestation, we used official remote sensing datasets to analyze deforestation inside and outside indigenous territories within Brazil's Amazon biome during the 2013-2021 period. Deforestation has increased by 129% inside ITs since 2013, followed by an increase in illegal mining areas. In 2019-2021, deforestation was 195% higher and 30% farther from the borders towards the interior of indigenous territories than in previous years (2013-2018). Furthermore, about 59% of carbon dioxide (CO2) emissions within ITs in 2013-2021 (96 million tons) occurred in the last three years of analyzed years, revealing the magnitude of increasing deforestation to climate impacts. Therefore, curbing deforestation in indigenous territories must be a priority for the Brazilian government to secure these peoples' land rights, ensure the forests' protection and regulate the global climate.

Strategies to increase HIV testing among men who have sex with men and transgender women: an integrative review.

BACKGROUND: Men who have sex with men (MSM) and transgender women (TGW) are disproportionately affected by HIV, with much higher incidence and prevalence rates than in the general population in different countries. There are several barriers to testing among MSM and TGW, such as low risk perception, anticipation of HIV-related stigma, discrimination of sexual orientation, in addition to difficulties related to care and access to health services. Therefore, analyzing the available evidence of the effectiveness of strategies for scaling up HIV testing among key populations is essential to point out potential knowledge gaps which may need to be addressed and develop public health policies to promote testing and early diagnosis of HIV infection. METHODS: An integrative review was carried out to evaluate strategies for scaling up HIV testing in these populations. Search strategy was performed on eight electronic databases, without language restriction. We included clinical trials, quasi-experimental studies, and non-randomized studies. Study selection and data extraction were both performed independently by pairs and disagreements were solved by a third revisor. The screening of the studies was carried out through the selection of titles/abstracts and the reading of the full texts of the pre-selected studies based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Data extraction was performed using a structured form. RESULTS: Thirty-seven publications referring to 35 studies were included, mostly being carried out in the United States of America and Australia. No studies were found evaluating disaggregated data on TGW. The studies were grouped into four types of intervention strategies: self-test distribution system (n = 10), organization of health services (n = 9), peer education (n = 6), and social marketing campaign (n = 10). Strategies that focused on the first three groups, combined or not, were more effective in increasing HIV testing among MSM. CONCLUSIONS: Considering the diversity of interventions and the methodological heterogeneity of the included studies, strategies especially involving self-test distribution systems, associated with new information and communication technologies, should be evaluated in different communities and social contexts. Research evaluating specific studies on TGW population is still needed.

Germline pathogenic variants in patients with early-onset neuroendocrine neoplasms.

Neuroendocrine neoplasms (NENs) are a rare group of cancers with heterogeneous behaviour and mostly of unknown aetiology. Excluding some infrequent hereditary cancer syndromes, the extent and clinical significance of mutations in other cancer predisposing genes (CPGs) are not known. We aimed to investigate the frequency of pathogenic and likely germline pathogenic variants (GPVs) in known CPGs in young adults with NEN and the clinical and molecular characteristics of these patients. We recruited 108 patients with lung or digestive NEN diagnosed between 18 and 50 years and performed targeted sequencing of 113 CPGs on germline DNA. For some patients, tumour features such as loss of heterozygosity (LOH), tumour mutation burden and microsatellite instability were evaluated. GPVs were detected in 17 patients (15.7%). Median age, sex, stage at diagnosis, family history of NENs or any personal history of neoplasm were similar between patients with or without GPVs. GPV carriers had more gastric (P = 0.084), functioning NEN (P = 0.041), positive family history of cancer (P = 0.015) and exclusively well-differentiated histology. Genes affected were mostly involved in DNA repair (CHEK2, ERCC2, ERCC3, XPC, MSH6, POLE and SLX4), with most GPVs found in MUTYH (four cases). LOH was performed in eight tumours and detected only in an SLX4-positive case. Overall, our findings indicate a role of inherited genetic alterations, particularly in DNA repair genes, in NEN carcinogenesis in young adults. These patients more often had a family history of cancer and functioning NENs.

The globalizability of temporal discounting.

Economic inequality is associated with preferences for smaller, immediate gains over larger, delayed ones. Such temporal discounting may feed into rising global inequality, yet it is unclear whether it is a function of choice preferences or norms, or rather the absence of sufficient resources for immediate needs. It is also not clear whether these reflect true differences in choice patterns between income groups. We tested temporal discounting and five intertemporal choice anomalies using local currencies and value standards in 61 countries (N = 13,629). Across a diverse sample, we found consistent, robust rates of choice anomalies. Lower-income groups were not significantly different, but economic inequality and broader financial circumstances were clearly correlated with population choice patterns.

Neutrophil-Derived COX-2 has a Key Role during Inflammatory Hyperalgesia.

Inflammation is a vital process for the injured tissue restoration and one of its hallmarks is inflammatory hyperalgesia. The cyclooxygenase (COX) pathway is strongly related to the inflammatory and painful process. Usually, the COX-1 isoform is described as homeostatic, while COX-2 is characterized as inducible in inflammatory conditions. Although it is well known that neutrophil cells are the first to arrive at the inflamed site and the major source of COX-2 is still unknown, the specific role of neutrophil-derived COX-2 in the pain process is. Thus, in the present study, we demonstrate for the first time that neutrophil-derived COX-2 plays a key role in peripheral inflammatory hyperalgesia. Conditional knockout mice for COX-2 in neutrophils (COX-2 fl/fl: Mrp8cre±) exhibited higher pain sensitivity after carrageenan (CG) injection and long-lasting IL-1ß-induced hyperalgesia compared with the control group (COX-2 fl/fl). Also, CG-induced inflammation in COX-2 fl/fl: Mrp8cre± mice showed COX-1 overexpression, and increased neutrophil migration and pro-inflammatory cytokines (e.g., IL-1ß and CXCL1). These findings revealed that neutrophil COX-2 has an important role in the regulation of inflammatory hyperalgesia.

Evaluation of phenotypic methods for detection of polymyxin B-resistant bacteria.

Determination of sensitivity to polymyxins has always been a challenge, especially in clinical laboratory routines. This study evaluated two rapid, simple, and inexpensive phenotypic methods to test polymyxin B (PMB) susceptibility in Enterobacterales and non-fermenting Gram-negative bacilli. One hundred isolates were used in the tests. The isolates were collected in three hospitals in southern and southeastern Brazil from 1995 to 2019. We compared broth microdilution (reference method) with the broth disk elution test and modified drop test, using polymyxin B -disk or PMB -powder in 2 concentrations (12 and 16 µg/ml). For the broth disk elution and modified drop test with the concentration of 12 µg/ml, categorical agreement values exceeded 90%. The modified drop test with a concentration of 12 µg/ml and broth disk elution may be excellent for initial screening of polymyxin-resistance in laboratory routines. Moreover, these methods are simple and use inexpensive supplies, and may optimize therapeutic decisions.

#Clareamentoalaser: características das postagens em português no Instagram® / #Clareamentoalaser: characteristics of posts in Portuguese on Instagram®

Objetivo: O objetivo deste estudo foi descrever e avaliar o conteúdo das publicações realizadas usando a hashtag #clareamentoalaser na rede social Instagram®. Materiais e métodos: Foram avaliadas 622 postagens relacionadas ao procedimento de clareamento dentário com uso de luz/laser, publicadas no primeiro trimestre do ano de 2021 (janeiro a março). As características analisadas foram o tipo de perfil, gênero, opinião exposta no conteúdo da publicação, tipo de conteúdo exposto na publicação, nomenclatura do perfil e número de curtidas. A análise foi realizada através do teste qui-quadrado de Pearson, sendo estipulado um nível de significância considerando p-valor <0,05. Resultados:Dentre as 497 publicações incluídas, 77,26% mostraram-se favoráveis ao uso de luz/laser no clareamento. Os valores apurados na variável número de curtidas variam de 0 a 3.361, com média de 42,56 por publicação. Em relação ao tipo de perfil, 60,16% eram cirurgiões-dentistas e 37,83% empresas ou clínicas odontológicas, sendo o conteúdo da publicação majoritariamente comercial (75,65%). Ainda, 53,12% dos perfis foram identificados como do gênero feminino, 9,05% do gênero masculino e 37,83% outros, como empresas ou clínicas odontológicas. Mais de 70% das publicações feitas por profissionais, perfis leigos e empresas ou clínicas odontológicas foram favoráveis ao uso da luz/laser no clareamento dental (p< 0,0001). Discussão: Este estudo expressa a necessidade de educação em saúde, educação continuada e da prática odontológica ba-seada em evidências. Conclusão: Na rede social do Instagram®, houve predominância na divulgação de publicações favoráveis quanto a utilização da luz/laser no clareamento dentário, considerando o período do primeiro trimestre de 2021.

Aim: The aim of this study was to describe and assess the content of publications made using the hashtag #claramentoalaser on the social network Instagram®. Materials and methods: A total of 622 posts related to the tooth whitening procedure with the use of light/laser, published in the first quarter of 2021 (January to March) were evaluated. The characteristics analyzed were the type of profile, gender, opinion exposed in the content of the publication, content exposed in the publication, profile nomenclature and number of likes. The analysis was performed using Pearson's chi-square test, with a significance level considering p-value <0.05. Results: Among the 497 publications included, 77.26% had a positive opinion about the use of light/laser for dental bleaching. The values found in the variable number of likes range from 0 to 3,361, with an average of 42.56 per publication. Regarding the type of profile, 60.16% were dentists, 37.83% com-panies or dental clinics, with commercial content exposed in most publications (75.65%). Still, 53.12% of the profiles were identified as female, 9.05% male and 37.83% others, such as companies or dental clinics. More than 70% of publications made by dentists, lay profiles, and companies or dental clinics had a positive opinion about light/laser use in bleaching (p<0.0001). Discussion: This study expresses the need to adopt an evidence-based practice, health education and continuing education in dentistry. Conclusions: In the Instagram® social network, was identified predominance of publications with a positive opinion about the use of light/laser in tooth whitening in 2021, first quarter.

The non-explicit observational method is reproducible and valid in the analysis of occupational biomechanical exposure of workers.

BACKGROUND: An analysis of workers' exposure to biomechanical risk factors in the workplace with a high methodological quality allows for stronger evidence of the relationship between risk factors and musculoskeletal disorders. OBJECTIVE: To evaluate the measurement properties of non-explicit observational methods. METHODS: Psychometric study design that included 50 workers in an analysis of occupational tasks (n = 50 video tasks) using a non-explicit observational method, the Rapid Entire Body Assessment (REBA) method, and a workers' self-report on their perception of their biomechanical occupational exposure. The reliability was tested by a non-explicit method and the REBA method. The validity of the non-explicit method was tested using the REBA method and the workers' self-report, according to COSMIN guidelines. RESULTS: The intra- and inter-evaluator reliability of the non-explicit observational method ranged from moderate to strong (ICC2.1  = 0.45 to 0.87), and the agreement was acceptable (SEM = 0.60 to 2.79), expect for neck region. The REBA method showed intra-evaluator reliability to be moderate to excellent (ICC2.1  = 0.50 to 1.00), inter-evaluator reliability to be poor to strong (ICC2.1  = -0.17 to 0.83), and intra- and inter-evaluator agreement to be poor to moderate (SEM = 0.00 to 0.74). The concurrent validity between the non-explicit and REBA methods showed a strong positive correlation. CONCLUSION: The non-explicit observational method of biomechanical exposure analysis presented reproducible and valid measurement properties. Methods that present better measurement properties provide professionals with a better basis for analysis and assist in decision-making to reduce or eliminate risk exposure.

Investigação de causas genéticas associadas ao desenvolvimento de sarcomas em idade jovem / Investigation of genetic causes associated with the development of sarcomas at young age

Os avanços recentes na genômica permitiram o reconhecimento de quase uma centena de novos genes de predisposição ao câncer (GPC). Embora o rastreamento genético de uma parte destes genes esteja atualmente bem estabelecido para os tumores hereditários mais comuns, há uma série de tumores raros, como sarcomas, que podem estar associados a síndromes de câncer hereditário, mas cuja relação genótipo-fenótipo ainda é desconhecida. Dessa forma, o presente estudo propôs-se a: definir a frequência de variantes raras germinativas patogênicas em GPCs em crianças, adolescentes e adultos jovens com sarcomas; avaliar as características moleculares dos tumores para confirmar sua causa genética em casos selecionados, realizando análise de perda de heterozigose (LOH); identificar novos genes e vias relevantes associados ao desenvolvimento de condrossarcomas e sarcomas ultrarraros. Para isso, foram rastreadas variantes germinativas em 177 pacientes jovens diagnosticados com sarcoma abaixo de 40 anos, através da análise de sequenciamento de nova geração, com painéis customizados de 113/126 GPCs. Primeiramente, foi rastreada a variante fundadora do gene TP53 mais prevalente na população brasileira (p.Arg337His) e 5/177 pacientes foram detectados como portadores e excluídos da análise de painel. No painel, foram avaliados 172 pacientes e em 33 foram detectadas variantes patogênicas (P) ou provavelmente patogênicas (PP) (18,6%), sendo estas em genes previamente associados ao risco no desenvolvimento de sarcomas, como CHEK2, EXT1, EXT2, NF1, RB1 e TP53, mas também em genes no qual esse risco ainda é desconhecido (AKT1, ERCC3, ERCC4, FANCM, MITF, MUTYH, NTHL1, SLX4 e TSC2) ou é emergente (BRCA2, ERCC2, PALB2). Cento e trinta e nove (78,5%) pacientes tiveram variantes de significado incerto ou foram negativos. Portadores de variantes P/PP foram mais propensos a apresentar mais de um tumor primário do que não portadores (21,1% X 6,5%; p=0,012). Dos 25 tumores avaliados para LOH, a perda do alelo selvagem foi detectada em nove (36%), sendo um dos genes de associação desconhecida a sarcomas (MITF). Para explorar novos genes candidatos, 9 pacientes com condrossarcomas ou sarcomas ultrarraros negativos para variantes P/PP foram incluídos em análise de sequenciamento de exoma tumoral e germinativo. Foram priorizadas na análise germinativa variantes em 10 GPCs conhecidos e 22 genes com variantes de perda de função e missenses em que a relação com câncer hereditário é desconhecida. Foi encontrada somente uma variante em um gene associado a sarcomas (RECQL4) e em um gene emergente (BRCA2). Uma variante patogênica no gene RAD50 foi detectada em um paciente com sarcoma fibromixóide. Em dois tumores distintos, foi observada alteração no gene COL7A1. Na análise somática, em três genes (TP53, PTCH1, CREBBP) foram encontradas alterações potencialmente significantes clinicamente, visto que são genes de associação conhecida a sarcomas. Também foram identificados outros genes mutados, alguns deles incluídos em vias biológicas que podem ser interessantes para sarcomas, como VPS16 e MYF6. Contudo, para que seja possível realizar associações genótipofenótipo dos dados de exoma, tanto germinativo quanto somático, outras evidências são necessárias, como análise de transcriptoma e co-segregação. Com os dados do painel, nossos resultados destacam uma alta taxa de variantes P/PP em GPCs em pacientes jovens brasileiros com sarcoma (21,5% incluindo os pacientes portadores da variante TP53 p.Arg337His), mesmo em pacientes sem história familiar de câncer. As taxas de variantes P/PP variaram de 0% a 33% entre os subtipos de sarcoma e exibiram associações específicas entre subtipos e genes. Isso aponta a urgência de implementar estratégias de triagem genética adequadas para esses indivíduos e suas famílias.

Recent advances in genomics have allowed the recognition of nearly a hundred new cancer predisposing genes (CPGs). Although genetic screening in some of these genes is currently well established for the most common hereditary tumors, there are a number of rare tumors, such as sarcomas, that may be associated with hereditary cancer syndromes, but whose genotypephenotype relationship is still unknown. Thus, the present study aimed to: define the frequency of rare pathogenic germline variants in CPGs in children, adolescents and young adults with sarcomas; assess the molecular characteristics of tumors to confirm their genetic cause in selected cases, performing loss of heterozygosity (LOH) analysis; identify novel genes and relevant pathways associated with the development of chondrosarcomas and ultra-rare sarcomas. For this, we searched for germline variants in 177 young patients diagnosed with sarcoma under 40 years old, through next-generation sequencing analysis, with customized panels of 113/126 CPGs. First, we searched for the founder variant in TP53 gene (p.Arg337His) - most prevalent in the Brazilian population - and found that 5/177 patients were detected as carriers, being excluded from the panel analysis. In the panel, 172 patients were evaluated and in 33 (18.6%) there were detected germline pathogenic variants (GPVs), occurring in genes previously associated with the risk of developing sarcomas (TP53, RB1, NF1, CHEK2, EXT1 and EXT2), but also in genes where that risk is still unknown (ERCC2/3, TSC2, RAD50, FANCM, and others) or is emerging (PALB2, BRCA2). GPVs carriers were more likely to present more than one primary tumor than non-carriers (21.1% X 6.5%; p=0.012). One hundred and thirty-nine (78.5%) patients had variants of uncertain significance or were negative. Of the 25 tumors evaluated for LOH, loss of the wild-type allele was detected in nine (36%), in which one of the genes has an unknown association with sarcomas (MITF). To explore new candidate genes, 9 patients with chondrosarcomas or ultra-rare sarcomas negative for GPVs were included in tumor exome and germline sequencing analysis. In the germline analysis, there were prioritized variants in 10 known CPGs and 22 genes with loss-of-function and missense variants in which the relationship with hereditary cancer is unknown. Only one variant was found in a sarcoma-associated gene (RECQL4) and in an emergent gene (BRCA2). A pathogenic variant in the RAD50 gene was detected in a patient with fibromyxoid sarcoma. An alteration in COL7A1 gene was observed in two different tumors. In the somatic analysis, there were found potentially clinically significant alterations in three genes (TP53, PTCH1, CREBBP), since they are genes with a known association with sarcomas. Other mutated genes were also identified, some of them included in biological pathways that may be interesting for sarcomas, such as VPS16 and MYF6. However, in order to be able to perform genotype-phenotype associations of exome data, other evidences are needed for both germline and somatic evaluation, such as transcriptome analysis and co-segregation. With panel data, our results highlight a high rate of GPVsin CPGs in young Brazilian patients with sarcoma (21.5% including patients carrying the TP53:p.Arg337His variant), even in patients with no family history of cancer. Our findings reveal that 1 in each 5 patients with sarcomas under 40 years old presented a GPV, even in patients without family history of cancer. GPVs rates varied from 0% to 33% across sarcoma subtypes and exhibited specific gene-subtypes associations. This pinpoints the urgency of implementing appropriated genetic screening strategies for these individuals and their families.

Internações de idosos por condições crônicas sensíveis à atenção primárianuma região do Ceará / Hospitalizations of older adults due to chronic conditions sensitive to primary care in a Ceará region

Objetivo:analisar a evolução temporal das ICSAP crônicas em idosos e sua correlação com a cobertura da ESF e indicadores socioeconômicos de uma região de saúde. Método: estudo ecológico descritivo, cuja unidade de análise foram os municípios da 11ª Região de Saúde do Cearáno período de 2012 a 2017. Os dados foram extraídos do Sistema de Informação Hospitalar, Sistema de Informação e-Gestor e Atlas de Desenvolvimento Humano no Brasil e analisados no software STATA. Resultados: registradas 7.245 ICSAP,4.650 (64,18%) pertencem aos grupos de condições crônicas. A maior frequência de internações foi entre os idosos do sexo masculino 2.578(55,4%) e com idade de 70 a 74 anos com 1.873 (35,87%). As causas mais prevalentes foram: Insuficiência Cardíaca com 1.666(35,83%), DoençasCerebrovasculares com 1.048(22,54%) e Angina com 754 (16,22%). Houve tendência de redução das ICSAP crônicas e expansão da cobertura da ESF com forte correlação inversa (r= -0,80, p-valor =0,0301) e com o índice de Gini (R = 0,80, p =0,0291). Os demais indicadores socioeconômicos não apresentaram correlação com as taxas de ICSAP. Conclusão: a expansão da cobertura da ESF esteve fortemente correlacionada à redução das taxas de ICSAP crônicasem idosos. Assim, é oportuno exortar profissionais e gestores para desenvolver estratégias no território que favoreça um cuidado efetivo.

Objective:to analyze the time evolution of chronic HPCSCs in older adults and its correlation with the FHS coverage and socioeconomic indicators of a health region. Method:a descriptive and ecological study, whose unit of analysis were the municipalities of the 11thHealth Region of Ceará during the 2012-2017period. The data were extracted from the Hospital Information System, e-Gestor Information System and Atlas of Human Development in Brazil, and analyzed in the STATAsoftware.Results: a total of 7,245HPCSCswere recorded, of which 4,650(64.18%) belonged to the chronic condition groups. The highest frequency of hospitalizations was among male older adults, with 2,578(55.4%), and aged from 70to74years old, with 1,873(35.87%). The most prevalent causes werethe following: Heart Failure with 1,666(35.83%), Cerebrovascular Diseases with 1,048(22.54%), and Angina with 754(16.22%). There was a tendency towards a reduction of chronic HPCSCs and expansion of the FHS coverage with a stronginverse correlation(r=-0.80,p-value=0.0301), as well as with the Gini index(R=0.80, p=0.0291). The other socioeconomic indicators showed no correlation with the HPCSC rates. Conclusion: the expansion of the FHS coverage was strongly correlated with a reduction in therates of chronic HPCSCs in older adults. Thus, it is appropriate to urge professionals and managers to develop strategies in the territory that favor effective care

Resistant hypertension: Prevalence and profile of patients followed in a university ambulatory.

BACKGROUND: Hypertension affects about 36 million Brazilians. It is estimated that 10%-20% of these have resistant hypertension. These patients are at an increased risk of early target organ damage, as well as cardiovascular and renal events. OBJECTIVE: To estimate the prevalence of resistant hypertension in a specialized outpatient clinic, to describe the sociodemographic and clinical characteristics of these patients, and to identify possible factors associated with resistant hypertension. METHODS: Data collection from medical records of hypertensive patients treated using oral antihypertensive drugs in optimized doses at a specialized university clinic from March 2014 to December 2014, after ethical approval statement. All patients were using appropriate antihypertensive drugs in optimized doses and assisted at a teaching-assistance clinic of internal medicine of the Bahiana School of Medicine and Public Health in Brazil. RESULTS: A total of 104 patients were enrolled and 31.7% (n = 33) had criteria for resistant hypertension. Of the total participants, 75.7% were female and 54.8% were black or brown. The average age was 61.7 years (SD ± 10.1). In the resistant hypertension group, 63.6% had diabetes, compared to 32.4% in the hypertension group. Among resistant hypertensive patients, 51.5% had dyslipidemia. Regarding drug treatment, 75.8% of the resistant hypertension group and 51.4% of the hypertension group used statins. Among patients with resistant hypertension, 90.9% used angiotensin II receptor blockers and 66.7%, dihydropyridine calcium channel blockers. In the resistant hypertension group, 75.8% used beta-blockers, against 25.4% in the hypertension group. CONCLUSION: The prevalence of hypertension was higher than that described in the global literature, which may be associated with the high percentage of black and brown ("pardos") patients in the population studied, and also because the study was performed in a specialized outpatient clinic.

Lipid nanoparticles loaded with butamben and designed to improve anesthesia at inflamed tissues.

The most frequently used local anesthetics (LA) for local infiltration have an ionizable amine in the range of pH 7.6-8.9. Effective anesthesia of inflamed tissues is a great challenge, especially because the induced local acidosis decreases the fraction of the neutral (more potent) LA species in situ. To solve this limitation, the butyl-substituted benzocaine analogue butamben (BTB) - that has no ionizable amine group close to the physiological pH - could be useful if it was not for its low solubility. To overcome the solubility problem, an optimized formulation for BTB using nanostructured lipid carriers (NLC) was developed by a factorial design and characterized using DLS, XRD, DSC and cryo-EM. The release kinetics and cytotoxicity of the new formulation were measured in vitro, while the in vivo tests assessed its effectiveness on healthy and inflamed tissues, in rats. The optimized NLCBTB formulation showed desirable physicochemical properties (size = 235.6 ± 3.9 nm, polydispersity = 0.182 ± 0.006 and zeta potential = -23.6 ± 0.5 mV), high (99.5%) encapsulation efficiency and stability during 360 days of storage at room temperature. NLCBTB prolonged the release of butamben and decreased its in vitro cytotoxicity without inducing any in vivo toxic alteration. In the inflammatory hyperalgesia model, the NLCBTB formulation showed potential for the management of inflammatory pain, displaying greater analgesic effectiveness (40%) and a prolonged effect.

Regional inequalities in cervical cancer survival in Minas Gerais State, Brazil.

BACKGROUND: Cervical cancer survival is marked by socioeconomic and demographic inequalities. We investigated differences in survival across health regions in Minas Gerais, Brazil, in cervical cancer patients who underwent treatment in the Brazilian Public Health System. METHODS: From a database developed through probabilistic and deterministic linkage of data from information systems of the Brazilian Public Health System, we identified cervical cancer cases, diagnosed between 2002 and 2010, who underwent radiation and/or chemotherapy and lived in Minas Gerais, Brazil. Five-year overall and cause-specific survivals were estimated by the Kaplan-Meier method and compared using the log-rank test. We used extended Cox models to assess the relationship between the health region of residence and the overall and cause-specific death risk, adjusting for relevant variables. RESULTS: We included 5613 patients with a median age of 55.0 years. Median follow-up time was 70.0 months. Five-year overall and cause-specific survivals were 56.3 % and 63.6 %, respectively. Across the 13 health regions, 5-year survival ranged from 46.6%-64.2% (p < 0.001) in the overall analysis and from 52.0% to 72.0% (p < 0.001) in the cause-specific analysis. Multivariate models revealed a significantly higher death risk for most health regions in comparison to the reference health region (Norte). Adjustment by age, tumor stage, comorbidity, treatment, travel time, and year of diagnosis had little effect on the association. CONCLUSION: We found regional disparities in cervical cancer survival that persisted after relevant adjustments. Uneven regional provision of health services might be implicated in these disparities, affecting timely access to treatment for cervical cancer patients.