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Abstract The synthetic lethal effect observed with the use of PARP inhibitors (PARPi) with tumors characterized by the loss of key players in the homologous recombination (HR) pathway, commonly referred to as “BRCAness”, is maintaining high interest in oncology. While BRCAness is a well-established feature in breast, ovarian, prostate, and pancreatic carcinomas, our recent findings indicate that up to 15% of colorectal cancers (CRC) also harbor defects in the HR pathway, presenting promising opportunities for innovative therapeutic strategies in CRC patients. We developed a new tool called HRDirect, which builds upon the HRDetect algorithm and is able to predict HR deficiency (HRD) from reference-free tumor samples. We validated HRDirect using matched breast cancer and CRC patient samples. Subsequently, we assessed its efficacy in predicting response to the PARP inhibitor olaparib by comparing it with two other commercial assays: AmoyDx HRD by Amoy Diagnostics and the TruSight Oncology 500 HRD (TSO500-HRD) panel by Illumina NGS technology. While all three approaches successfully identified the most PARPi-sensitive CRC models, HRDirect demonstrated superior precision in distinguishing resistant models compared to AmoyDX and TSO500-HRD, which exhibited overlapping scores between sensitive and resistant cells. Furthermore, we propose integrating HRDirect scoring with ATM and RAD51C immunohistochemical analysis as part of our “composite biomarker approach” to enhance the identification of HRD tumors, with an immediate translational and clinical impact for CRC personalized treatment.

Abstract Background Liquid biopsy based on cell-free DNA (cfDNA) analysis holds significant promise as a minimally invasive approach for the diagnosis, genotyping, and monitoring of solid malignancies. Human tumors release cfDNA in the bloodstream through a combination of events, including cell death, active and passive release. However, the precise mechanisms leading to cfDNA shedding remain to be characterized. Addressing this question in patients is confounded by several factors, such as tumor burden extent, anatomical and vasculature barriers, and release of nucleic acids from normal cells. In this work, we exploited cancer models to dissect basic mechanisms of DNA release. Methods We measured cell loss ratio, doubling time, and cfDNA release in the supernatant of a colorectal cancer (CRC) cell line collection (N = 76) representative of the molecular subtypes previously identified in cancer patients. Association analyses between quantitative parameters of cfDNA release, cell proliferation, and molecular features were evaluated. Functional experiments were performed to test the impact of modulating DNA methylation on cfDNA release. Results Higher levels of supernatant cfDNA were significantly associated with slower cell cycling and increased cell death. In addition, a higher cfDNA shedding was found in non-CpG Island Methylator Phenotype (CIMP) models. These results indicate a positive correlation between lower methylation and increased cfDNA levels. To explore this further, we exploited methylation microarrays to identify a subset of probes significantly associated with cfDNA shedding and derive a methylation signature capable of discriminating high from low cfDNA releasers. We applied this signature to an independent set of 176 CRC cell lines and patient derived organoids to select 14 models predicted to be low or high releasers. The methylation profile successfully predicted the amount of cfDNA released in the supernatant. At the functional level, genetic ablation of DNA methyl-transferases increased chromatin accessibility and DNA fragmentation, leading to increased cfDNA release in isogenic CRC cell lines. Furthermore, in vitro treatment of five low releaser CRC cells with a demethylating agent was able to induce a significant increase in cfDNA shedding. Conclusions Methylation status of cancer cell lines contributes to the variability of cfDNA shedding in vitro. Changes in methylation pattern are associated with cfDNA release levels and might be exploited to increase sensitivity of liquid biopsy assays.

Abstract Background Despite recent therapeutic advances, combating cancer resistance remains a formidable challenge. The 78-kilodalton glucose-regulated protein (GRP78), a key stress-inducible endoplasmic reticulum (ER) chaperone, plays a crucial role in both cancer cell survival and stress adaptation. GRP78 is also upregulated during SARS-CoV-2 infection and acts as a critical host factor. Recently, we discovered cardiac glycosides (CGs) as novel suppressors of GRP78 stress induction through a high-throughput screen of clinically relevant compound libraries. This study aims to test the possibility that agents capable of blocking stress induction of GRP78 could dually suppress cancer and COVID-19. Results Here we report that oleandrin (OLN), is the most potent among the CGs in inhibiting acute stress induction of total GRP78, which also results in reduced cell surface and nuclear forms of GRP78 in stressed cells. The inhibition of stress induction of GRP78 is at the post-transcriptional level, independent of protein degradation and autophagy and may involve translational control as OLN blocks stress-induced loading of ribosomes onto GRP78 mRNAs. Moreover, the human Na+/K+-ATPase α3 isoform is critical for OLN suppression of GRP78 stress induction. OLN, in nanomolar range, enhances apoptosis, sensitizes colorectal cancer cells to chemotherapeutic agents, and reduces the viability of patient-derived colon cancer organoids. Likewise, OLN, suppresses GRP78 expression and impedes tumor growth in an orthotopic breast cancer xenograft model. Furthermore, OLN blocks infection by SARS-CoV-2 and its variants and enhances existing anti-viral therapies. Notably, GRP78 overexpression mitigates OLN-mediated cancer cell apoptotic onset and suppression of virus release. Conclusion Our findings validate GRP78 as a target of OLN anti-cancer and anti-viral activities. These proof-of-principle studies support further investigation of OLN as a readily accessible compound to dually combat cancer and COVID-19.

Multiple strategies are continuously being explored to expand the drug target repertoire in solid tumors. We devised a novel computational workflow for transcriptome‐wide gene expression outlier analysis that allows the systematic identification of both overexpression and underexpression events in cancer cells. Here, it was applied to expression values obtained through RNA sequencing in 226 colorectal cancer (CRC) cell lines that were also characterized by whole‐exome sequencing and microarray‐based DNA methylation profiling. We found cell models displaying an abnormally high or low expression level for 3533 and 965 genes, respectively. Gene expression abnormalities that have been previously associated with clinically relevant features of CRC cell lines were confirmed. Moreover, by integrating multi‐omics data, we identified both genetic and epigenetic alternations underlying outlier expression values. Importantly, our atlas of CRC gene expression outliers can guide the discovery of novel drug targets and biomarkers. As a proof of concept, we found that CRC cell lines lacking expression of the MTAP gene are sensitive to treatment with a PRMT5‐MTA inhibitor (MRTX1719). Finally, other tumor types may also benefit from this approach.

Background: Inhibitor eradication to restore factor (F)VIII efficacy is the treatment goal for persons with severe hemophilia A (HA) and inhibitors. Immune tolerance induction (ITI) is demanding and successful in about 70% of people. Until now, it has remained difficult to quantify the probability of ITI success or failure, complicating the decision to initiate or not initiate ITI. Estimating the individual chance of ITI success allows clinicians, patients, and their families to support shared decision-making. Objectives: We aimed to identify clinical predictors of ITI success and to develop a clinical prediction model to estimate the chance of successful ITI in persons with severe HA. Methods: This multicenter study included persons with severe HA who received ITI. Clinical data were collected. Successful ITI was defined by a negative inhibitor titer and an adequate response to FVIII concentrates. A multivariable logistic regression model was developed. Model performance and internal validation were performed. Results: Of 206 participants with a median age of 19.8 months (IQR, 12.1-38.8) at ITI start, 148 (71.8%) achieved ITI success. Our clinical prediction model included 4 predictors of ITI success: cumulative number of FVIII exposure days at inhibitor development, peak inhibitor titer, ethnicity, and F8 mutation type. The C statistic was 0.801 (95% CI, 0.70-0.87). Conclusion: In our study, including 206 people with severe HA and inhibitors, we developed a clinical prediction model to estimate the chance of successful ITI. After future external validation, this clinical prediction model may be useful for informing clinicians and families.

Background: Eradicating inhibitors to restore the effectiveness of Factor VIII (FVIII) is a desirable treatment goal for People with Severe Hemophilia A (PwSHA) and inhibitors, as this enables treatment of bleeding episodes with FVIII concentrates. However, since Immune Tolerance Induction (ITI) is a burdensome and costly treatment, it is important to identify determinants for ITI success to decide if it is worth undertaking ITI. For more effective treatment allocation, there is a need to identify which persons will or will not benefit from ITI. Aim: We aimed to identify determinants of successful ITI in PwSHA and to use these determinants in a clinical prediction model. Methods: German, Brazilian, Dutch, Canadian, and Italian PwSHA who underwent ITI were included. The primary outcome was clinical ITI success, defined by (1) a negative inhibitor titer, and (2) an adequate clinical response to FVIII concentrates. Clinical determinants included F8 genotype, race, age, cumulative exposure days to FVIII, inhibitor titers, and ITI regimen. Genetic determinants included in the analyses were FCGR gene, IL10 CA short tandem repeat, and gene variants. Crude Relative Risks (RR) were calculated for ITI success with 95% Confidence Intervals (95% CI). Results: In total, 224 PwSHA were included (61 German, 51 Brazilian, 45 Dutch, 39 Canadian, and 28 Italian). The median ages at inhibitor development and ITI start were 2-years (Interquartile Range [IQR 1‒3]) and 2-years (IQR 1‒6). The median interval between inhibitor diagnosis and ITI start was 17-weeks (IQR 2‒64). Most ITI trials started with recombinant FVIII (130; 58%) and the median prescribed regimen at ITI start was 43 IU/kg/day (IQR 21‒200). The need for adjuvant therapy and central venous access infection were related to failure, while inhibitor titer at detection < 10 BU/mL, inhibitor titer immediately before ITI start < 10 BU/mL, and peak inhibitor titer ever measured < 100 BU/mL were related to success. Inhibitor peak titer below 100 BU/mL showed the highest chance for ITI success (OR = 11.5, 95% CI 5.5‒24.3, p < 0.001). Conclusion: Historical peak titer below 100 BU/mL was the strongest determinant for ITI success. Genetic analyses will be available by the due date, and we will develop a prediction model to estimate the chance of success.

Immune Tolerance Induction (ITI) is the only treatment to eradicate inhibitors in people with Severe Hemophilia A (SHA). Successful ITI restores Factor VIII (FVIII) tolerance. ITI is demanding and successful in approximately 70% of people. Therefore, identifying predictors of ITI outcome is essential to guide clinical decision-making. We aimed to identify genetic predictors of ITI success in people with SHA and inhibitors who underwent ITI. This observational multicenter study included people with SHA who underwent ITI, between 2015 and 2023. Clinical and patient data including factor VIII gene (F8) mutation type and DNA samples were collected. Successful ITI was defined by a negative inhibitor titer and an adequate response to FVIII concentrates. The associations between ITI success and F8 genotype and 216 candidate predictors including single nucleotide polymorphisms (SNPs) and human leukocyte antigen (HLA)-variants employing a global screening array (GSA), CA dinucleotide Short Tandem Repeat (STR) polymorphisms in the Interleukin (IL)-10 promoter region, and FCGR2/3 gene locus variations were analyzed. Of 204 participants, 147 (72.1%) achieved ITI success. The majority (52.0%) of participants had F8 intron 22 inversion. None of the candidate SNPs/HLA-variants, IL-10 CA dinucleotide STR, or FCGR2/3 gene locus variations were associated with ITI success. F8 large deletions were negatively associated with ITI success (OR = 0.15, 95% CI 0.04‒0.51, p = 0.002). Our study including 204 people with SHA identified F8 large deletions as a predictor of ITI failure. Pooling cohorts may allow the identification of additional genetic predictors of ITI success in the future.

Introdução: A dengue é uma das doenças virais transmitidas por vetores mais importantes da atualidade. O Brasil vive sua maior epidemia da doença, onde a infecção pelo vírus da dengue (DENV) causa doenças que variam amplamente em severidade, desde dengue febril auto-limitada a dengue hemorrágica com risco de morte. Além dos efeitos diretos do vírus, tem sido proposta imunopatogênese associada a auto-anticorpos, onde a infecção viral da dengue induz anticorpos autoreativos contra plaquetas, células do endotélio e moléculas da coagulação, podendo também estar associada a outros mecanismos. Caso clínico: KCLTS, 29 anos, primigesta, entra em trabalho de parto por bolsa rota, parto normal. RN com 2,680 kg, icterícia neonatal zona 4-5, apresentando Hb = 18,7 g/dL, bilirrubina total 26,88 mg/dL, bilirrubina indireta 25,20 mg/dL. Solicitado exsanguíneo transfusão e RN encaminhado para fototerapia dupla. RN A RhD positivo, TAD positvo 3+, mãe O RhD positivo PAI positiva 2+ em ambas as células de triagem. Painel Liss/Coombs demonstrou anticorpo reativo 2+ em todas as células, inclusive autocontrole, negativando em painel enzimático. A gestante nunca havia recebido transfusão. Painel do eluato da amostra do RN todo negativo. Diante destes resultados realizado auto-adsorção com hemácias da mãe tratadas por bromelina. Após 3 adsorções realizou-se novamente o painel de identificação de anticorpos, que se mostrou negativo em todas as células confirmando a presença de auto-anticorpos. Após pesquisa de patologias pregressas ao parto, soube-se que a gestante estava positiva para o vírus da dengue 8 dias antes do parto, além de diabetes gestacional, sem outras comorbidades. Não foi possível determinar o subtipo do vírus. Optou-se pelo cancelamento da transfusão e manutenção da fototerapia além da administração de imunoglobulina endovenosa 1g/kg. RN teve alta em 16 dias do nascimento com bilirrubina total 5,61 mg/dL e hematócito de 30,10 %. Conclusão: A icterícia apresentada pelo RN provavelmente se deve aos autoanticorpos formados pela mãe devido à infecção pelo vírus da dengue.

Abstract The aim of this study was to evaluate the impact of oral conditions and health-related quality of life (HRQoL) on oral health-related quality of life (OHRQoL) in children and adolescents with blood coagulation disorders and hemoglobinopathies (BCDH). The study was cross-sectional and included 61 individuals aged 2 to 18 years with BCDH. Exams for dental caries (dmft/DMFT index), oral hygiene (simplified oral hygiene index – OHI-S), and gingival health (modified gingival index – MGI) were performed. The pediatric quality of life inventory™ (PedsQL™) generic core scale and oral health scale were used to measure HRQoL and OHRQoL. Spearman’s correlation coefficient (ρ) and the Mann-Whitney test (α = 0.05) were conducted to assess the relationship between covariates and the PedsQL™ oral health scale. The mean PedsQL™ oral health scale score was 76.66 (SD = 21.36). Worse OHRQoL was correlated with poor oral hygiene (ρ = -0.383; p: 0.004), poor gingival health (ρ = -0.327; p = 0.014), and better HRQoL (ρ = 0.488; p < 0.001). Greater untreated dental caries experience was associated with worse OHRQoL (p = 0.009). Worse oral health status in children and adolescents with BCDH negatively impacts OHRQoL, and OHRQoL and quality of life analyzed from a generic perspective are positively correlated constructs in this population.

Introduction: Treatment of hemophilia A in Brazil is offered to all patients at no cost. However, several unmet medical needs exist. Method: In this study, we applied the Delphi method to discuss with seven hemophilia A specialists the challenges that patients and the health system face regarding hemophilia A treatment and opportunities for improvement. Results: A consensus was obtained regarding the number of weekly infusions and patient adherence to treatment. The bleeding profile, unfavourable pharmacokinetics (PKs), low adherence and high daily activity were patient profiles that would benefit from using the extended half-life (EHL) recombinant factor VIII (rFVIII). The advantages of treatment with the EHL rFVIII were the lower number of infusions per week, which could increase patient adherence and decrease the risk of bleeds, due to a more constant plasma level, a lower value. Additionally, the EHL rFVIII could improve quality of life, especially in patients with high daily activity, such as adolescents and young adults. The panelists mentioned that EHL rFVIII, if available, could be offered first to the priority group (adolescents between 12 and 19 years old), followed by adults (20 to 64 years old) and elderly people (over 65 years old). Conclusion: In summary, the EHL rFVIII offers the optimal prophylaxis by decreasing the dose frequency, increasing the treatment adherence and improving the QoL, without compromising safety and efficacy.

Succession following the abandonment of traditional management practices can pose severe consequences for the conservation of semi-natural dry grassland communities. In the present study, we investigated whether the quantity of floral resources changes during succession of semi-natural dry grasslands and how this is related to pollinator richness and the number of pollination interactions at the community level. We addressed this issue by quantifying floral resources (i.e., number of flowers, nectar volume and number of pollen grains) and monitoring pollination interactions in dry grassland communities at different stages of succession, defined as the total cover of plant species of forest edges. The relationship between the quantity of floral resources and cover of plant species of forest edges was significantly hump-shaped, i.e., regardless of the type of floral resource, all peaked at intermediate values of cover of plant species of forest edges. The richness of animal-pollinated plants in bloom also showed a hump-shaped relationship with the cover of plant species of forest edges, while the richness of pollinator species and the number of pollination contacts were indirectly related to the cover of plant species of forest edges, as they were significantly associated with the number of flowers and the richness of animal-pollinated plants in bloom. Results suggest that succession of dry grasslands after abandonment may affect a crucial function in terrestrial ecosystems, namely animal-mediated pollination. Nevertheless, the conditions of early succession, which could be achieved by the presence of scattered shrubs, could ultimately be favourable for the pollination function in dry grasslands.

Roots play a fundamental role in forest ecosystems, but obtaining samples from deep layers remains a challenging process due to the methodological and financial efforts required. In our quest to understand the dynamics of Eucalyptus roots, we raise three fundamental questions. First, we inquire about the average extent of the roots of two contrasting Eucalyptus genotypes. Next, we explore the factors that directly influence the growth and depth of these roots, addressing elements such as soil type, climate, and water availability. Lastly, we investigate how the variation in Eucalyptus species may impact root growth patterns, biomass, and carbon stock. In this study, we observed that the maximum root depth increased by an average of 20% when genotypes were grown on sites with higher water availability (wet site). E. urophylla stands had a higher biomass and carbon stock (5.7 Mg C ha−1) of fine roots when cultivated on dry sites (annual rainfall~727 mm) than the wet sites (annual rainfall~1590 mm). In E. grandis × E. camaldulensis stands, no significant differences were observed in the stock of fine root biomass (3.2 Mg C ha−1) between the studied environments. Our results demonstrated that genotypes with greater drought tolerance (E. grandis × E. camaldulensis) tend to maintain higher stocks of fine root biomass (3.2–6.3 Mg ha−1) compared to those classified as plastic (E. urophylla), regardless of the edaphoclimatic conditions of the cultivation site. Finally, our research helps understand how Eucalyptus trees adapt to their environment, aiding sustainable forest management and climate change mitigation. We also provide a practical tool to estimate underground biomass, assisting forest managers and policymakers in ensuring long-term forest sustainability.

Introdução/Objetivos: A Imunotolerância (IT) é o tratamento padrão para erradicar aloanticorpos neutralizantes contra o fator VIII (FVIII) em pessoas com hemofilia A (PcHA). A IT foi implementada como política pública no Brasil em 2011. Até o presente, nenhum estudo avaliou os custos do consumo de fator de coagulação com as doses preconizadas pelo protocolo brasileiro de IT. O objetivo deste estudo foi descrever os custos da IT de acordo com o seu desfecho. Método: Estudo de coorte retrospectivo, conduzido em 4 Centros de Hemofilia, com dados de janeiro/2011 a julho/2021. As PcHA foram tratadas conforme protocolo brasileiro de IT (50 unidades internacionais de FVIII/kilograma (kg), 3x por semana). Foram avaliados o custo dos fatores de coagulação em 3 períodos: (i) pré-IT (12 meses antes do início da IT); (ii) durante a IT (período variável) e (iii) pós-IT (12 meses após seu término) e analisados conforme seu desfecho (sucesso total, sucesso parcial ou falha). Os custos foram corrigidos pelo Índice Nacional de Preços ao Consumidor Amplo (IPCA/junho de 2022), ajustados pela paridade do dólar americano (purchasing power parity – PPP dólar/2022) e avaliados com nível de significância de 0,05. Resultados: Foram incluídas 91 PcHA, das quais 87 (95,60%) eram graves (FVIII

Objetivo: Descrever um caso de manejo de pseudotumor de retroperitônio em paciente com Hemofilia A grave. Caso clínico: Paciente masculino, 52 anos, portador de Hemofilia A grave em uso de profilaxia com concentrado de Fator VIII plasmático 2500UI 3 vezes na semana, procura atendimento por hematoma volumoso em região femoral direita. Exame de ressonância magnética realizado há 2 meses mostrava hematoma expansivo no ventre do iliopsoas se estendendo ao compartimento anterior da coxa com volume estimado de 1760 cm3. Evoluiu sem melhora no período, com nova ressonância magnética mostrando lesão expansiva, heterogênea, lobulada e multisseptada se estendendo da fossa ilíaca direita até a diáfise distal do fêmur direito, com volume estimado de 5113 cm3, compatível com pseudotumor. Paciente internado, iniciada reposição de FVIII 1000UI a cada 6 horas. Após 6 semanas a ressonância magnética foi repetida e mostrou persistência da lesão, agora com volume de 4574 cm3. Solicitada pesquisa de inibidor de FVIII que resultou negativa. Decidido por tratamento com embolização de ramo circunflexo superficial da artéria ilíaca externa direita como tentativa de conter o sangramento. Devido a persistência da lesão, optado por ressecção cirúrgica do pseudotumor. Ressecção de pseudotumor realizada pela ortopedia sem sangramento importante durante o procedimento. Pseudotumor de 3300 g ressecado inteiro, hemostasia realizada com selante de fibrina. Paciente evoluiu com reabilitação com fisioterapia e hoje possui membro funcionante em uso de profilaxia com FVIII. Discussão: Pseudotumor é uma complicação infrequente porém grave em pacientes hemofílicos. A incidência estimada é de 1-2% dos pacientes com Hemofilia grave, chegando a 10% em paciente com inibidor de FVIII. Consiste na formação de uma lesão cística resultante de sangramentos recorrentes em tecidos moles ou ósseos, e se não tratado pode evoluir com calcificação e ossificação, levando a destruição óssea, articular e fraturas patológicas. Não há consenso sobre o tratamento, sendo descrito na literatura diferentes modalidades terapêuticas de tratamento cirúrgico, conservador, embolização e radioterapia. A abordagem cirúrgica é necessária para pseudotumores grandes, profundos e com mais tempo de evolução, apesar da alta incidência de complicações como sangramento perioperatório, formação de fístula, infecção, desenvolvimento de inibidor e necessidade de amputação. Sua realização depende de uma equipe multidisciplinar experiente. Conclusão: Relatamos um caso raro de um grande pseudotumor em paciente com Hemofilia A grave e os desafios de seu manejo.

Introdução/Objetivos: Ainda não é possível identificar quais pacientes com hemofilia A (HA) desenvolverão inibidor. O objetivo desse estudo foi construir um modelo de predição de desenvolvimento inibidores usando uma rede de variáveis através da inteligência artificial (IA). Métodos: Crianças com HA (CHA) moderada/grave (FVIII5UB). O algoritmo identificou associação entre as variáveis e o desfecho, das quais linfócitos TCD4, linfócitos B ativos, monócitos, micropartícula derivada de monócito e IL17 foram as mais relevantes. Os valores preditivos positivo e negativo do modelo foram, respectivamente, de 74,2% e 98.4%. O recall foi de 95,8% e o F1-score foi de 83,7%. O modelo foi capaz de identificar corretamente o desfecho de todas as crianças com robustez acima de 23,5%. Ao restringir a análise aos pacientes com mutações de alto risco, a acurácia para identificar, em T0, as CHA que desenvolveram inibidor, aumentou de 74% para 82,1%. Os resultados derivados da validação cruzada leave-one-out demonstraram a capacidade da rede de prever com precisão o desenvolvimento do inibidor. Discussão: Este modelo de IA avaliou diferentes variáveis, agrupando as CHA de acordo com o perfil de redes em comum e demonstrou que indivíduos que desenvolvem inibidores exibem padrões de classificação distintos em comparação com aqueles que não os desenvolvem. Este modelo foi capaz de identificar diferenças topológicas nas redes conforme os perfis clínicos, genéticos e imunológicos de CHA em T0. A acurácia do modelo para identificar crianças que desenvolveram inibidor aumentou após considerar as mutações de alto risco. Conclusão: Nosso algoritmo identificou, ao diagnóstico da HA, uma diferença entre a rede de variáveis em CHA moderada/grave que desenvolveram e que não desenvolveram inibidor, com uma acurácia média de 90.5%.

O Hemocentro Coordenador do Paraná possui equipe de triadores composta por médicos e enfermeiros qualificados, capacitados e conhecedores das regras previstas na legislação vigente. O grupo se reúne a cada seis meses, presencialmente e on-line , com o intuito de discutir ocorrências de inaptidões não normatizadas na legislação ministerial, atualizar o Manual de Triagem Clínica e adequar protocolos às determinações de auditorias internas e externas. A equipe conta com assessoria de convidados especialistas de áreas de conhecimento como Endocrinologia, Odontologia, Infectologia e outros para esclarecimentos de eventos pontuais. Foram temas recentemente discutidos e elucidados: cirurgia bariátrica, uso de testosterona em candidatos trans, enxerto ósseo bovino e contato domiciliar nas Hepatites B e C. Os temas a serem discutidos são enviados previamente à equipe no grupo de WhatsApp “Apoio Triadores Hemepar”. No caso de não haver consenso, ou persistirem dúvidas, a demanda é enviada à responsável técnica institucional, que compõe a equipe, para esclarecimento posterior ou inclusão na pauta da reunião seguinte. Com as normatizações institucionais publicadas no Manual de Triagem Clínica, as Ouvidorias com reclamações geradas por doadores diminuíram consideravelmente, promovendo melhora nos indicadores institucionais. Os critérios gerais para a doação de sangue aprovados pela equipe de triadores são disponibilizados para as unidades da hemorrede paranaense no site da Secretaria de Estado da Saúde, no link https://www.saude.pr.gov.br/Pagina/Doacao-de-Sangue . Nesse momento, os candidatos à doação têm a oportunidade de conhecer alguns dos principais critérios com antecedência, evitando deslocamentos que geram custo e frustração, desnecessariamente.

A hemofilia A é um distúrbio hemorrágico caracterizado pela deficiência ou anormalidade da atividade coagulante do fator VIII. Uma de suas complicações é o desenvolvimento de inibidor que trata-se de um anticorpo capaz de inibir a ação coagulante do fator em reposição, o que resulta em um tratamento não responsivo e no aumento das intercorrências hemorrágicas. A alternativa para desensibilização do indivíduo aloimunizado e erradicação do inibidor contra o fator VIII é o protocolo de imunotolerância (IT). Entretanto, 20 a 40% dos casos podem não responder à indução de IT, demandando um tratamento menos eficaz com o uso contínuo de agentes de bypassing. Em 2021, no Brasil, foi aprovada uma nova alternativa terapêutica para a pessoa com hemofilia A e inibidor com falha na IT, qual seja o protocolo para uso de emicizumabe. Esse anticorpo monoclonal mimetiza a ação do fator VIII ativado na cascata de coagulação e promove a hemostasia. Sendo assim, o objetivo desse estudo é relatar o caso de um adolescente (P1) com hemofilia A grave e inibidor persistente após IT após um 20 meses em tratamento com emicizumabe com vistas às melhorias de sua condição de saúde. O método é descritivo-exploratório com base nos registros da equipe multidisciplinar obtidos no prontuário e através de entrevista aberta ao paciente e seu responsável. Para essa análise, os dados de sangramento foram analisados no período de outubro de 2020 até julho de 2023, o que corresponde há um ano antes do início do emicizumabe até o presente momento. P1, sexo masculino, 15 anos, HAG, diagnosticado no primeiro ano de vida, fez uso de fator VIII por demanda. Desenvolveu inibidor no quinto ano de vida, realizou tratamento de IT (2014-2018) com falha terapêutica e profilaxia de longa duração com agente de bypassing. Da análise dos registros da avaliação musculo-esquelética antes do tratamento com emicizumabe consta: auxílio de órtese externa para mobilidade; artrofia grau IV na musculatura de membro inferior esquerdo; sinovite crônica em joelho esquerdo com piora da perimetria e também sinovite em joelho e cotovelo direito. Apresentou hemartroses recorrentes no período, com taxa anual de sangramento = 7 e três hospitalizações, sendo duas delas para tratamento hospitalar de sangramento episódico e outra para sinovectomia artroscópica de joelho esquerdo. Apresentava problemas com autoestima, isolamento social e absenteísmo escolar. A falta de registros no diário de infusão também sugeriam uma má adesão ao tratamento proposto. A partir do uso do anticorpo monoclonal, em outubro de 2021, não houve sangramentos espontâneos (taxa sangramento anual = 0); melhora significativa da sinovite de ambos os joelhos com diminuição do flexo do joelho esquerdo e deambulação sem auxílio. Nos demais aspectos relacionados a saúde, evidenciou-se a retomada da rotina escolar e das atividades sociais e de lazer, com percepções positivas acerca da sua autoimagem. Dessa forma, sugere-se que o impacto do tratamento atual na saúde articular e a diminuição da frequência de aplicação das doses profiláticas, além da via de administração, foram facilitadores para o desenvolvimento do auto-cuidado e para um melhor engajamento do indivíduo e sua família em relação ao tratamento proposto. Além disso, a redução da taxa de sangramento é um achado que corrobora com os resultados de eficácia da droga publicados nos estudos recentes.

Resumo Introdução Inúmeras pessoas são afetadas por transtornos mentais ao longo da vida, acarretando diminuição da expectativa de vida saudável e redução da saúde geral. Objetivo Descrever a prevalência de transtornos mentais e sua associação com variáveis demográficas, econômicas, comportamentais e de saúde de usuários da Atenção Primária à Saúde (APS). Método Estudo transversal realizado em 35 unidades básicas de saúde da zona urbana da cidade de Pelotas-RS. Foram entrevistados 525 pessoas com 18 anos ou mais. O desfecho do estudo foi os transtornos mentais, verificado pela seguinte questão: “Em alguma consulta, algum médico lhe disse que o Sr.(a) possui alguma doença? Se sim, quais?”. Considerou-se com transtorno mental os usuários que relataram as seguintes doenças: depressão, ansiedade, transtorno bipolar, problemas psicológicos e problemas emocionais. Resultados A prevalência de transtornos mentais foi de 12,1%. A depressão foi a doença mais relatada pelos usuários (74,6%). Indivíduos fumantes apresentaram prevalência de transtornos mentais duas vezes maior quando comparados a indivíduos que nunca fumaram. Pessoas que ingeriam três ou mais medicamentos de maneira contínua tinham probabilidade de possuir transtornos mentais sete vezes maior do que os indivíduos que não utilizavam medicamentos. Conclusão Apresentaram transtornos mentais 12,1% dos usuários das unidades básicas de saúde, sendo a depressão a doença mais prevalente. Uso de medicamentos de forma contínua e fumo foram associadas à maior prevalência de transtornos mentais.

Abstract Telomere maintenance is necessary to maintain cancer cell unlimited viability. However, the mechanisms maintaining telomere length in colorectal cancer (CRC) have not been extensively investigated. Telomere maintenance mechanisms (TMM) include the re-expression of telomerase or alternative lengthening of telomeres (ALT). ALT is genetically associated with somatic alterations in alpha-thalassemia/mental retardation X-linked (ATRX) and death domain-associated protein (DAXX) genes. Cells displaying ALT present distinctive features including C-circles made of telomeric DNA, long and heterogenous telomeric tracts, and telomeric DNA co-localized with promyelocytic leukemia (PML) bodies forming so-called ALT-associated PML bodies (APBs). Here, we identified mutations in ATRX and/or DAXX genes in an extensive collection of CRC samples including 119 patient-derived organoids (PDOs) and 232 established CRC cell lines. C-circles measured in CRC PDOs and cell lines showed low levels overall. We also observed that CRC PDOs and cell lines did not display a significant accumulation of APBs or long telomeres with no appreciable differences between wild-type and mutated ATRX/DAXX samples. Overall, our extensive analyses indicate that CRC is not prone to engage ALT, even when carrying genetic lesions in ATRX and/or DAXX, and support the notion that ATRX/DAXX genomic footprints are not reliable predictors of ALT.

Abstract Phishing is an attack characterized by attempted fraud against users. The attacker develops a malicious page that is a trusted environment, inducing its victims to submit sensitive data. There are several platforms, such as PhishTank and OpenPhish, that maintain databases on malicious pages to support anti-phishing solutions, such as, for example, block lists and machine learning. A problem with this scenario is that many of these databases are disorganized, inconsistent, and have some limitations regarding integrity and balance. In addition, because phishing is so volatile, considerable effort is put into preserving temporal information from each malicious page. To contribute, this article built a phishing database with consistent and balanced data, temporal information, and a significant number of occurrences, totaling 942,471 records over the 5 years between 2016 and 2021. Of these records, 135,542 preserve the page’s source code, 258,416 have the attack target brand detected, 70,597 have the hosting service identified, and 15,008 have the shortener service discovered. Additionally, 123,285 records store WHOIS information of the domain registered in 2021. The data is available on the website https://piracema.io/repository.

Abstract N-acylhydrazones are considered privileged structures in medicinal chemistry, being part of antimicrobial compounds (for example). In this study we show the activity of N-acylhydrazone compounds, namely AH1, AH2, AH4, AH5 in in vitro tests against the chloroquine-resistant strain of Plasmodium falciparum (W2) and against WI26 VA-4 human cell lines. All compounds showed low cytotoxicity (LC50 > 100 µM). The AH5 compound was the most active against Plasmodium falciparum, with an IC50 value of 0.07 μM. AH4 and AH5 were selected among the tested compounds for molecular docking calculations to elucidate possible targets involved in their mechanism of action and the SwissADME analysis to predict their pharmacokinetic profile. The AH5 compound showed affinity for 12 targets with low selectivity, while the AH4 compound had greater affinity for only one target (3PHC). These compounds met Lipinski's standards in the ADME in silico tests, indicating good bioavailability results. These results demonstrate that these N-acylhydrazone compounds are good candidates for future preclinical studies against malaria. Graphical Abstract