Interações complexas na hipercolesterolemia familiar: um estudo de caso de aterosclerose multivascular e variantes genéticas / Complex interactions in familial hypercholesterolemia: a case study of multivascular atherosclerosis and genetic variants

INTRODUÇÃO: A hipercolesterolemia familiar (HF) é um distúrbio genético bem reconhecido que se manifesta como níveis significativamente elevados de LDL-c sérico devido a aberrações no metabolismo das lipoproteínas. A maioria dos casos de HF, especificamente 85-90%, está associada a mutações patogênicas no gene do receptor de LDL (LDLR). Além disso, a presença de mutações de perda de função (LOF) no gene PCSK9 tem sido associada a um risco reduzido de doença cardiovascular, um fenômeno observado mesmo quando variantes patogênicas de LDLR coexistem. MÉTODOS: Este estudo incluiu um homem de 46 anos com dislipidemia e uso inconsistente de rosuvastatina. Na história familiar consta uma irmã (caso índice) e um sobrinho diagnosticados com HF portadores de variante patogênica de LDLR (c.313+1G>A, heterozigoto). RESULTADOS: O paciente relatou o aparecimento de dor precordial típica um mês antes da consulta. O exame físico revelou arco corneano. Os resultados laboratoriais iniciais mostraram colesterol total em 254 mg/dL, LDL-c em 191 mg/dL, HDL-c em 49 mg/dL e triglicerídeos em 69 mg/dL. Um escore Dutch MedPed de 11 confirmou HF. A ecocardiografia transtorácica indicou função ventricular esquerda normal. A angiografia coronária revelou calcificação significativa e estenose da artéria coronária direita (RCA) e da artéria circunflexa esquerda proximal (LCx). A intervenção incluiu angioplastia LCx e manejo conservador da RCA. Medicamentos pós-alta incluíram AAS 100mg/dia, Clopidogrel 75mg/dia, Rosuvastatina 40mg/ dia e Ezetimibe 10mg/dia. O nível de LDL-c diminuiu para 76 mg/dL no acompanhamento. A análise genética confirmou a variante familiar de LDLR e identificou uma mutação LOF coexistente de PCSK9 (R46L, heterozigoto). CONCLUSÃO: Este caso destaca a gravidade e complexidade da aterosclerose multivascular associada à HF. Embora a variante LOF de PCSK9 seja potencialmente protetora, sua influência nos resultados clínicos permanece incerta. Essa ambiguidade reforça a progressão multifatorial da aterosclerose e a resposta variável ao tratamento em pacientes com HF. Isso ressalta a importância de identificar fatores genéticos e não genéticos adicionais que contribuem para a doença, além dos loci genéticos conhecidos para entender e gerenciar melhor essa condição clínica.

Unraveling the role of miRNAs as biomarkers in Chagas cardiomyopathy: Insights into molecular pathophysiology

BACKGROUND: Chagas cardiomyopathy (ChCM) is a severe form of Chagas disease and a major cause of cardiovascular morbidity and mortality. The dysregulation of the immune response leads to cardiac remodeling and functional disruptions, resulting in life-threatening complications. Conventional diagnostic methods have limitations, and therapeutic response evaluation is challenging. MicroRNAs (miRNAs), important regulators of gene expression, show potential as biomarkers for diagnosis and prognosis. AIM: This review aims to summarize experimental findings on miRNA expression in ChCM and explore the potential of these miRNAs as biomarkers of Chagas disease. METHODS: The search was conducted in the US National Library of Medicine MEDLINE/PubMed public database using the terms "Chagas cardiomyopathy" OR "Chagas disease" AND "microRNA" OR "miRNA" OR "miR." Additionally, bioinformatics analysis was performed to investigate miRNA-target interactions and explore enrichment pathways of gene ontology biological processes and molecular functions. RESULTS: The miR-21, miR-146b, miR-146a, and miR-155 consistently exhibited up-regulation, whereas miR-145 was down-regulated in ChCM. These specific miRNAs have been linked to fibrosis, immune response, and inflammatory processes in heart tissue. Moreover, the findings from various studies indicate that these miRNAs have the potential as biomarkers for the disease and could be targeted in therapeutic strategies for ChCM. CONCLUSION: In this review, we point out miR-21, miR-146b, miR-146a, miR-155, and miR-145-5p role in the complex mechanisms of ChCM. These miRNAs have been shown as potential biomarkers for precise diagnosis, reliable prognostic evaluation, and effective treatment strategies in the ChCM.

Unraveling the role of miRNAs as biomarkers in Chagas cardiomyopathy: Insights into molecular pathophysiology.

BACKGROUND: Chagas cardiomyopathy (ChCM) is a severe form of Chagas disease and a major cause of cardiovascular morbidity and mortality. The dysregulation of the immune response leads to cardiac remodeling and functional disruptions, resulting in life-threatening complications. Conventional diagnostic methods have limitations, and therapeutic response evaluation is challenging. MicroRNAs (miRNAs), important regulators of gene expression, show potential as biomarkers for diagnosis and prognosis. AIM: This review aims to summarize experimental findings on miRNA expression in ChCM and explore the potential of these miRNAs as biomarkers of Chagas disease. METHODS: The search was conducted in the US National Library of Medicine MEDLINE/PubMed public database using the terms "Chagas cardiomyopathy" OR "Chagas disease" AND "microRNA" OR "miRNA" OR "miR." Additionally, bioinformatics analysis was performed to investigate miRNA-target interactions and explore enrichment pathways of gene ontology biological processes and molecular functions. RESULTS: The miR-21, miR-146b, miR-146a, and miR-155 consistently exhibited up-regulation, whereas miR-145 was down-regulated in ChCM. These specific miRNAs have been linked to fibrosis, immune response, and inflammatory processes in heart tissue. Moreover, the findings from various studies indicate that these miRNAs have the potential as biomarkers for the disease and could be targeted in therapeutic strategies for ChCM. CONCLUSION: In this review, we point out miR-21, miR-146b, miR-146a, miR-155, and miR-145-5p role in the complex mechanisms of ChCM. These miRNAs have been shown as potential biomarkers for precise diagnosis, reliable prognostic evaluation, and effective treatment strategies in the ChCM.

Outcomes of preoperative chemotherapy for colorectal cancer with peritoneal metastasis underwent cytoreductive surgery.

BACKGROUND: This study aims to assess and compare the extent to which preoperative chemotherapy prior to CRS improves survival in patients diagnosed with CRCPM. METHODS: We included 251 patients from 2012 to 2019 in our center. Inverse probability of treatment weighting (IPTW) analysis was used to minimize the selection bias. Survival analysis was performed to compare the survival outcomes. Multivariate Cox regression analysis was conducted to identify prognostic factors. RESULT: The baseline characteristics were well balanced using IPTW (standardized mean difference < 0.1). Preoperative chemotherapy cannot significantly improve overall survival (HR, 1.03; 95% CI 0.71-1.49; P = 0.88). In subgroup analysis, we found that intestinal obstruction after preoperative chemotherapy significantly reduced survival (HR, 2.25; 95% CI 1.01-5.03; P = 0.048), while in the upfront surgery group, intestinal obstruction had no impact on prognosis. CONCLUSION: For CRCPM patients treated with CRS, preoperative chemotherapy does not seem to prolong overall survival. Furthermore, the emergence of intestinal obstruction after chemotherapy may compromise the effectiveness of treatment, resulting in a worse prognosis. This finding has important clinical implications for treatment decisions.

Mitochondrial DNA haplogroups and variants predispose to chagas disease cardiomyopathy

Cardiomyopathies are major causes of heart failure. Chagas disease (CD) is caused by the parasite Trypanosoma cruzi, and it is endemic in Central and South America. Thirty percent of cases evolve into chronic chagas cardiomyopathy (CCC), which has worse prognosis as compared with other cardiomyopathies. In vivo bioenergetic analysis and ex vivo proteomic analysis of myocardial tissues highlighted worse mitochondrial dysfunction in CCC, and previous studies identified nuclear-encoded mitochondrial gene variants segregating with CCC. Here, we assessed the role of the mitochondrial genome through mtDNA copy number variations and mtDNA haplotyping and sequencing from heart or blood tissues of severe, moderate CCC and asymptomatic/indeterminate Chagas disease as well as healthy controls as an attempt to help decipher mitochondrial-intrinsic genetic involvement in Chagas disease development. We have found that the mtDNA copy number was significantly lower in CCC than in heart tissue from healthy individuals, while blood mtDNA content was similar among asymptomatic Chagas disease, moderate, and severe CCC patients. An MtDNA haplogrouping study has indicated that African haplogroups were over represented in the Chagas subject groups in comparison with healthy Brazilian individuals. The European lineage is associated with protection against cardiomyopathy and the macro haplogroup H is associated with increased risk towards CCC. Using mitochondria DNA sequencing, 84 mtDNA-encoded protein sequence pathogenic variants were associated with CCC. Among them, two variants were associated to left ventricular non-compaction and two to hypertrophic cardiomyopathy. The finding that mitochondrial protein-coding SNPs and mitochondrial haplogroups associate with risk of evolving to CCC is consistent with a key role of mitochondrial DNA in the development of chronic chagas disease cardiomyopathy.

Prenatal ethanol exposure and changes in fetal neuroendocrine metabolic programming.

Prenatal ethanol exposure (PEE) (mainly through maternal alcohol consumption) has become widespread. However, studies suggest that it can cause intrauterine growth retardation (IUGR) and multi-organ developmental toxicity in offspring, and susceptibility to various chronic diseases (such as neuropsychiatric diseases, metabolic syndrome, and related diseases) in adults. Through ethanol's direct effects and its indirect effects mediated by maternal-derived glucocorticoids, PEE alters epigenetic modifications and organ developmental programming during fetal development, which damages the offspring health and increases susceptibility to various chronic diseases after birth. Ethanol directly leads to the developmental toxicity of multiple tissues and organs in many ways. Regarding maternal-derived glucocorticoid-mediated IUGR, developmental programming, and susceptibility to multiple conditions after birth, ethanol induces programmed changes in the neuroendocrine axes of offspring, such as the hypothalamus-pituitary-adrenal (HPA) and glucocorticoid-insulin-like growth factor 1 (GC-IGF1) axes. In addition, the differences in ethanol metabolic enzymes, placental glucocorticoid barrier function, and the sensitivity to glucocorticoids in various tissues and organs mediate the severity and sex differences in the developmental toxicity of ethanol exposure during pregnancy. Offspring exposed to ethanol during pregnancy have a "thrifty phenotype" in the fetal period, and show "catch-up growth" in the case of abundant nutrition after birth; when encountering adverse environments, these offspring are more likely to develop diseases. Here, we review the developmental toxicity, functional alterations in multiple organs, and neuroendocrine metabolic programming mechanisms induced by PEE based on our research and that of other investigators. This should provide new perspectives for the effective prevention and treatment of ethanol developmental toxicity and the early prevention of related fetal-originated diseases.

De novo purine metabolism is a metabolic vulnerability of cancers with low p16 expression.

p16 is a tumor suppressor encoded by the CDKN2A gene whose expression is lost in ~50% of all human cancers. In its canonical role, p16 inhibits the G1-S phase cell cycle progression through suppression of cyclin dependent kinases. Interestingly, p16 also has roles in metabolic reprogramming, and we previously published that loss of p16 promotes nucleotide synthesis via the pentose phosphate pathway. Whether other nucleotide metabolic genes and pathways are affected by p16/CDKN2A loss and if these can be specifically targeted in p16/CDKN2A-low tumors has not been previously explored. Using CRISPR KO libraries in multiple isogenic human and mouse melanoma cell lines, we determined that many nucleotide metabolism genes are negatively enriched in p16/CDKN2A knockdown cells compared to controls. Indeed, many of the genes that are required for survival in the context of low p16/CDKN2A expression based on our CRISPR screens are upregulated in p16 knockdown melanoma cells and those with endogenously low CDKN2A expression. We determined that cells with low p16/Cdkn2a expression are sensitive to multiple inhibitors of de novo purine synthesis, including anti-folates. Tumors with p16 knockdown were more sensitive to the anti-folate methotrexate in vivo than control tumors. Together, our data provide evidence to reevaluate the utility of these drugs in patients with p16/CDKN2A-low tumors as loss of p16/CDKN2A may provide a therapeutic window for these agents.

Identification of pathogenic variants in the brazilian cohort with familial hypercholesterolemia using exon-targeted gene sequencing

Familial hypercholesterolemia (FH) is a monogenic disease characterized by high plasma low-density lipoprotein cholesterol (LDL-c) levels and increased risk of premature atherosclerotic cardiovascular disease. Mutations in FH-related genes account for 40% of FH cases worldwide. In this study, we aimed to assess the pathogenic variants in FH-related genes in the Brazilian FH cohort FHBGEP using exon-targeted gene sequencing (ETGS) strategy. FH patients (n = 210) were enrolled at five clinical sites and peripheral blood samples were obtained for laboratory testing and genomic DNA extraction. ETGS was performed using MiSeq platform (Illumina). To identify deleterious variants in LDLR, APOB, PCSK9, and LDLRAP1, the long-reads were subjected to Burrows-Wheeler Aligner (BWA) for alignment and mapping, followed by variant calling using Genome Analysis Toolkit (GATK) and ANNOVAR for variant annotation. The variants were further filtered using in-house custom scripts and classified according to the American College Medical Genetics and Genomics (ACMG) guidelines. A total of 174 variants were identified including 85 missense, 3 stop-gain, 9 splice-site, 6 InDel, and 71 in regulatory regions (3'UTR and 5'UTR). Fifty-two patients (24.7%) had 30 known pathogenic or likely pathogenic variants in FH-related genes according to the American College Medical and Genetics and Genomics guidelines. Fifty-three known variants were classified as benign, or likely benign and 87 known variants have shown uncertain significance. Four novel variants were discovered and classified as such due to their absence in existing databases. In conclusion, ETGS and in silico prediction studies are useful tools for screening deleterious variants and identification of novel variants in FH-related genes, they also contribute to the molecular diagnosis in the FHBGEP cohort.

Identification of pathogenic variants in the Brazilian cohort with Familial hypercholesterolemia using exon-targeted gene sequencing.

Familial hypercholesterolemia (FH) is a monogenic disease characterized by high plasma low-density lipoprotein cholesterol (LDL-c) levels and increased risk of premature atherosclerotic cardiovascular disease. Mutations in FH-related genes account for 40% of FH cases worldwide. In this study, we aimed to assess the pathogenic variants in FH-related genes in the Brazilian FH cohort FHBGEP using exon-targeted gene sequencing (ETGS) strategy. FH patients (n = 210) were enrolled at five clinical sites and peripheral blood samples were obtained for laboratory testing and genomic DNA extraction. ETGS was performed using MiSeq platform (Illumina). To identify deleterious variants in LDLR, APOB, PCSK9, and LDLRAP1, the long-reads were subjected to Burrows-Wheeler Aligner (BWA) for alignment and mapping, followed by variant calling using Genome Analysis Toolkit (GATK) and ANNOVAR for variant annotation. The variants were further filtered using in-house custom scripts and classified according to the American College Medical Genetics and Genomics (ACMG) guidelines. A total of 174 variants were identified including 85 missense, 3 stop-gain, 9 splice-site, 6 InDel, and 71 in regulatory regions (3'UTR and 5'UTR). Fifty-two patients (24.7%) had 30 known pathogenic or likely pathogenic variants in FH-related genes according to the American College Medical and Genetics and Genomics guidelines. Fifty-three known variants were classified as benign, or likely benign and 87 known variants have shown uncertain significance. Four novel variants were discovered and classified as such due to their absence in existing databases. In conclusion, ETGS and in silico prediction studies are useful tools for screening deleterious variants and identification of novel variants in FH-related genes, they also contribute to the molecular diagnosis in the FHBGEP cohort.

Correlação entre o escore SAME-TT2R2 e o polimorfismo VKORC1: C. 174-136C>T (RS9934438) como preditores da qualidade da anticoagulação entre pacientes idosos brasileiros usuários de varfarina / Correlation between the SAME-TT2R2 score and the VKORC1:C.174-136C>T (RS9934438) polymorphism as predictors of anticoagulation quality among elderly Brazilian patients using warfarin

INTRODUÇÃO: A varfarina é um anticoagulante utilizado na prevenção e tratamento de doenças tromboembólicas. Nos idosos, a principal indicação é fibrilação atrial. O exame mais utilizado para controle da anticoagulação é o tempo de protrombina, através do cálculo da razão normalizada internacional (RNI), na faixa 2-3, que requerem um monitoramento especial devido a idade, dificuldades na adesão, polimedicação ou comorbidades. Embora seja conhecido que os fatores genéticos influenciam na resposta terapêutica, na maioria dos hospitais a farmacogenética ainda não é considerada no cálculo de ajuste de dose. Portadores do genótipo rs9934438 AA requerem menor dose comparado com genótipo AG ou GG. O escore SAMe- -TT2R2 obtido por meio de algumas variáveis visa predizer quais os pacientes em uso de varfarina atingirão taxas de INR aceitáveis e, por consequência, um tempo na faixa terapêutica (TFT) adequado. Apostolakis, propuseram e validaram o escore SAMe-TT2R2 (S=sexo; A =idade OBJETIVO: Visando estabelecer uma conduta terapêutica personalizada, o presente estudo tem como objetivo avaliar correlação entre o polimorfismo rs9934438 do gene VKORC1 e correlaciona-lo com o escore SAME-TT2R2 na predição da qualidade da anticoagulação em uso de varfarina. MÉTODOS e RESULTADOS: Foram incluídos 29 pacientes, idade de 81,72 (±4,07), de ambos os sexos e em uso de varfarina. A análise do polimorfismofoi realizada através da PCR em tempo real utilizando os reagentes do kit TaqMan™ Sample-to-SNP™ e o sistema de detecção TaqMan® SNP Genotyping Assay. As análises estatísticas foram realizadas utilizando o pacote SPSS v. 16.0 e nível de significância adotado foi de 5% e o Escore SAME-TT2R2 foi calculado após entrevista. Os Escores Chadsvasc2 = 4,34 (±1,17), Hasbled = 2,24 (±0,68) e SAME-TT2R2 = 2,31(±1,28), a dose semanal variou de 10 a 55 mg. Apesar de ser um estudo piloto, com baixo amostra, a distribuição dos genótipos está em equilíbrio gênico, segundo Hardy-Weinberg (AA=24,3%, AG=41,3%, GG= 34,4%). Os portadores do genótipo AA necessitaram de menor dose para atingir a TFT quando comparados aos portadores do genótipo GG: 26,6 versus 39,7 mg/semana, corroborando o resultado do SAME-TT2R2 = 2 no grupo AA comparado ao GG que foi de 3. CONCLUSÃO: Os resultados mostram que o polimorfismo rs9934438 está associado com a dose semanal de varfarina administrada aos pacientes idosos de pelo menos 30% e com correspondência ao Escore SAME-TT2R2 que demostrou um índice menor.

Morphological Effects of Cigarette Smoking on Respiratory System and Related Functions Based on AQPs Expression

SUMMARY: A great deal of attention of air pollution on respiratory health is increasing, particularly in relation to haze days. It is that exposure to cigarette smoke augments the toxicity of common air contaminants, thereby increasing the complexity of respiratory diseases. Although there are various mechanisms involved to respiratory diseases caused or worsen by cigarette smoking, in which the role of AQPs in the lung with regard to fluid homeostasis still remains elusive. In this paper, we copied the rat models based on smoke generator, and investigated the morphological changes of mucosa and related functions depending on the balance of lining liquid of alveoli via AQPs expression. Compared with normal group, weak labelling of AQP1 and AQP5 protein abundance were clearly detected in the corresponding part of smoke exposure groups compared with normal group. Hence, it is suggested that the contribution of AQPs in the lung is diminished, thereby causing perturbed balancing between resorptive and secretory fluid homeostasis under cigarette smoking.

Cada vez se presta más atención a la contaminación del aire en la salud respiratoria, particularmente, en relación con los días de neblina. En consecuencia la exposición al humo del cigarrillo aumenta la toxicidad de los contaminantes comunes del aire, lo que además aumenta la complejidad de las enfermedades respiratorias. Aunque existen varios mecanismos involucrados en las enfermedades respiratorias causadas o empeoradas por el tabaquismo, en las que el papel de las AQP en el pulmón respecto a la homeostasis de líquidos sigue siendo difícil de alcanzar. En este artículo, copiamos los modelos de rata basados en el generador de humo e investigamos los cambios morfológicos de la mucosa y las funciones relacionadas según el equilibrio del líquido de revestimiento de los alvéolos a través de la expresión de AQP. En comparación con el grupo normal, se detectó claramente un etiquetado débil de la abundancia de proteínas AQP1 y AQP5 en la parte correspondiente de los grupos de exposición al humo en comparación con el grupo control. Por lo tanto, se sugiere que la contribución de las AQP en el pulmón está disminuida, provocando así un equilibrio perturbado entre la homeostasis del líquido secretor y de reabsorción bajo el hábito de fumar cigarrillos.

Longitudinal study comparing IgG antibodies induced by heterologous prime­boost COVID­19 vaccine

ABSTRACT: Vaccines are critical cost­effective tools to control the COVID­19 pandemic. The heterologous prime­boost vaccination has been used by many countries to overcome supply issues, so the effectiveness and safety of this strategy need to be better clarified. This study aims to verify the effect of heterologous prime­boost COVID­19 vaccination on healthcare professionals from Dante Pazzanese Hospital in Brazil. It was performed serological assays of vaccinated individuals after 2­dose of CoronaVac (Sinovac; n = 89) or ChAdOx1 nCoV­19 (Oxford­AstraZeneca; n = 166) followed by a BNT162b2 booster (Pfizer­BioNTech; n = 255). The serum antibodies anti­S (spike), anti­N (nucleocapsid), and anti­RBD (receptor binding domain) were assessed by enzyme­linked immunosorbent assay. The heterologous booster dose induced a 10­fold higher anti­Spike antibody regardless of the 2­dose of a prime vaccine. It was strikingly observed that BNT162b2 enhanced levels of anti­spike antibodies, even in those individuals who did not previously respond to the 2­dose of CoronaVac. In conclusion, the heterologous scheme of vaccination using mRNA as a booster vaccine efficiently enhanced the antibody response against SARS­CoV­2, especially benefiting those elderly who were seronegative with a virus­inactivated vaccine.

Longitudinal study comparing IgG antibodies induced by heterologous prime-boost COVID-19 vaccine.

Vaccines are critical cost-effective tools to control the COVID-19 pandemic. The heterologous prime-boost vaccination has been used by many countries to overcome supply issues, so the effectiveness and safety of this strategy need to be better clarified. This study aims to verify the effect of heterologous prime-boost COVID-19 vaccination on healthcare professionals from Dante Pazzanese Hospital in Brazil. It was performed serological assays of vaccinated individuals after 2-dose of CoronaVac (Sinovac; n = 89) or ChAdOx1 nCoV-19 (Oxford-AstraZeneca; n = 166) followed by a BNT162b2 booster (Pfizer-BioNTech; n = 255). The serum antibodies anti-S (spike), anti-N (nucleocapsid), and anti-RBD (receptor binding domain) were assessed by enzyme-linked immunosorbent assay. The heterologous booster dose induced a 10-fold higher anti-Spike antibody regardless of the 2-dose of a prime vaccine. It was strikingly observed that BNT162b2 enhanced levels of anti-spike antibodies, even in those individuals who did not previously respond to the 2-dose of CoronaVac. In conclusion, the heterologous scheme of vaccination using mRNA as a booster vaccine efficiently enhanced the antibody response against SARS-CoV-2, especially benefiting those elderly who were seronegative with a virus-inactivated vaccine.

Injury prevention in the training of triple jump athletes / Prevenção de lesões no treinamento de esportistas de salto triplo / Prevención de lesiones en el entrenamiento de atletas de triple salto

ABSTRACT Introduction: The triple jump technique is one of the techniques of the most difficult sport in athletics, requiring extremely high physical qualities such as muscular strength, explosive power, and coordination capacity of athletes. Training its young practitioners should prioritize the specific teaching of sports techniques and the prevention of injuries intrinsic to the sport. Objective: Promote preventive interventions in training young triple jump athletes. Methods: The research method was used with interviews with athletes, coaches, and specialists. A statistical analysis was performed to identify the most relevant injuries in the sport. A protocol of preventive activities to be added to the athletes' training was elaborated through the data obtained. Results: A survey of young triple jumpers in sports schools in some cities in Province A found that there were 62 junior triple jumpers, of which 48 had injuries, the injury rate was 77.4%, and the injury rate of junior triple jumpers was relatively high, mainly in the knee. After analyzing the factors that cause sports injuries, it is shown that by rationalizing the training plan for the triple jump and the scientific content of training, athletes gradually increase their adaptability and self-protective ability to the influence of external factors, mastering the methods of prevention and rehabilitation of sports injuries of the knee. Conclusion: The preventive interventions presented allow triple jump athletes to maintain their best competitive state and create competitive results at the professional level. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: A técnica do salto triplo é uma das técnicas esportivas mais difíceis do atletismo, requerendo qualidades físicas extremamente elevadas como força muscular, poder explosivo e capacidade de coordenação dos atletas. O treinamento de seus jovens praticantes deve priorizar tanto o ensino específico das técnicas esportivas quanto a prevenção de lesões intrínsecas do esporte. Objetivo: Fomentar intervenções preventivas no treinamento de jovens atletas praticantes de salto triplo. Métodos: Utilizou-se o método de pesquisa com entrevistas entre atletas, treinadores e especialistas. Uma análise estatística foi elaborada para levantar as lesões mais relevantes ao esporte. Através dos dados obtidos, foi elaborado um protocolo de atividades preventivas a serem adicionadas no treinamento dos esportistas. Resultados: Uma pesquisa com jovens saltadores triplos em escolas esportivas de algumas cidades da Província A constatou que havia 62 saltadores triplos juniores, dos quais 48 tinham lesões, a taxa de lesões era de 77,4%, e a taxa de lesões dos saltadores triplos juniores era relativamente alta, principalmente no joelho. Após analisar os fatores que causam lesões esportivas, mostra-se que, racionalizando o plano de treinamento do salto triplo e o conteúdo científico do treinamento, os atletas aumentam gradualmente sua adaptabilidade e capacidade de autoproteção à influência de fatores externos, dominando os métodos de prevenção e reabilitação das lesões esportivas do joelho. Conclusão: As intervenções preventivas apresentadas permitem que os atletas do salto triplo mantenham seu melhor estado competitivo e criem resultados de competição a nível profissional. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: La técnica del triple salto es una de las técnicas deportivas más difíciles del atletismo, ya que requiere cualidades físicas muy elevadas como la fuerza muscular, la potencia explosiva y la capacidad de coordinación de los atletas. La formación de sus jóvenes practicantes debe priorizar tanto la enseñanza específica de las técnicas deportivas como la prevención de las lesiones intrínsecas del deporte. Objetivo: Promover intervenciones preventivas en el entrenamiento de jóvenes atletas de triple salto. Métodos: Se utilizó el método de investigación con entrevistas a atletas, entrenadores y especialistas. Se elaboró un análisis estadístico para identificar las lesiones más relevantes en el deporte. A través de los datos obtenidos, se elaboró un protocolo de actividades preventivas a añadir en el entrenamiento de los deportistas. Resultados: Una encuesta realizada a jóvenes saltadores de triple en escuelas deportivas de algunas ciudades de la provincia A reveló que había 62 saltadores de triple junior, de los cuales 48 tenían lesiones, la tasa de lesiones era del 77,4%, y la tasa de lesiones de los saltadores de triple junior era relativamente alta, principalmente en la rodilla. Después de analizar los factores que causan las lesiones deportivas, se demuestra que al racionalizar el plan de entrenamiento del triple salto y el contenido científico del entrenamiento, los atletas aumentan gradualmente su capacidad de adaptación y autoprotección a la influencia de los factores externos, dominando los métodos de prevención y rehabilitación de las lesiones deportivas de rodilla. Conclusión: Las intervenciones preventivas presentadas permiten a los atletas de triple salto mantener su mejor estado competitivo y crear resultados de competición a nivel profesional. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Blood DNA methylation marks discriminate Chagas cardiomyopathy disease clinical forms

Chagas disease is a parasitic disease from South America, affecting around 7 million people worldwide. Decades after the infection, 30% of people develop chronic forms, including Chronic Chagas Cardiomyopathy (CCC), for which no treatment exists. Two stages characterized this form: the moderate form, characterized by a heart ejection fraction (EF) ≥ 0.4, and the severe form, associated to an EF < 0.4. We propose two sets of DNA methylation biomarkers which can predict in blood CCC occurrence, and CCC stage. This analysis, based on machine learning algorithms, makes predictions with more than 95% accuracy in a test cohort. Beyond their predictive capacity, these CpGs are located near genes involved in the immune response, the nervous system, ion transport or ATP synthesis, pathways known to be deregulated in CCCs. Among these genes, some are also differentially expressed in heart tissues. Interestingly, the CpGs of interest are tagged to genes mainly involved in nervous and ionic processes. Given the close link between methylation and gene expression, these lists of CpGs promise to be not only good biomarkers, but also good indicators of key elements in the development of this pathology.

Complement Is Required for Microbe-Driven Induction of Th17 and Periodontitis.

In both mice and humans, complement and Th17 cells have been implicated in periodontitis, an oral microbiota-driven inflammatory disease associated with systemic disorders. A recent clinical trial showed that a complement C3 inhibitor (AMY-101) causes sustainable resolution of periodontal inflammation, the main effector of tissue destruction in this oral disease. Although both complement and Th17 are required for periodontitis, it is uncertain how these immune components cooperate in disease development. In this study, we dissected the complement-Th17 relationship in the setting of ligature-induced periodontitis (LIP), a model that previously established that microbial dysbiosis drives Th17 cell expansion and periodontal bone loss. Complement was readily activated in the periodontal tissue of LIP-subjected mice but not when the mice were placed on broad-spectrum antibiotics. Microbiota-induced complement activation generated critical cytokines, IL-6 and IL-23, which are required for Th17 cell expansion. These cytokines as well as Th17 accumulation and IL-17 expression were significantly suppressed in LIP-subjected C3-deficient mice relative to wild-type controls. As IL-23 has been extensively studied in periodontitis, we focused on IL-6 and showed that LIP-induced IL-17 and bone loss required intact IL-6 receptor signaling in the periodontium. LIP-induced IL-6 was predominantly produced by gingival epithelial cells that upregulated C3a receptor upon LIP challenge. Experiments in human gingival epithelial cells showed that C3a upregulated IL-6 production in cooperation with microbial stimuli that upregulated C3a receptor expression in ERK1/2- and JNK-dependent manner. In conclusion, complement links the periodontal microbiota challenge to Th17 cell accumulation and thus integrates complement- and Th17-driven immunopathology in periodontitis.

Epigenetic regulation of transcription factor binding motifs promotes Th1 response in Chagas disease cardiomyopathy

Abstract: Chagas disease, caused by the protozoan Trypanosoma cruzi, is an endemic parasitic disease of Latin America, affecting 7 million people. Although most patients are asymptomatic, 30% develop complications, including the often-fatal Chronic Chagasic Cardiomyopathy (CCC). Although previous studies have demonstrated some genetic deregulations associated with CCCs, the causes of their deregulations remain poorly described. Based on bulk RNA-seq and whole genome DNA methylation data, we investigated the genetic and epigenetic deregulations present in the moderate and severe stages of CCC. Analysis of heart tissue gene expression profile allowed us to identify 1407 differentially expressed transcripts (DEGs) specific from CCC patients. A tissue DNA methylation analysis done on the same tissue has permitted the identification of 92 regulatory Differentially Methylated Regions (DMR) localized in the promoter of DEGs. An in-depth study of the transcription factors binding sites (TFBS) in the DMRs corroborated the importance of TFBS's DNA methylation for gene expression in CCC myocardium. TBX21, RUNX3 and EBF1 are the transcription factors whose binding motif appears to be affected by DNA methylation in the largest number of genes. By combining both transcriptomic and methylomic analysis on heart tissue, and methylomic analysis on blood, 4 biological processes affected by severe CCC have been identified, including immune response, ion transport, cardiac muscle processes and nervous system. An additional study on blood methylation of moderate CCC samples put forward the importance of ion transport and nervous system in the development of the disease.

Perfil de citocinas em biópsia miocárdica de paciente transplantado com COVID-19 / Cytokine profile in myocardial biopsy of a transplant patient with COVID-19

FUNDAMENTO: A inflamação pode estar envolvida na ativação imune do transplante, resultando em disfunção do enxerto. A COVID-19 é caracterizada pela liberação descontrolada e elevada de citocinas pró-inflamatórias e imunidade suprimida. Descrevemos um caso de paciente com transplante de coração e COVID-19 que desenvolveu disfunção do enxerto e os achados do perfil de citocinas na biopsia endomiocárdica. RELATO DE CASO: Paciente com transplante cardíaco há 13 anos apresentou-se no PS com queixa de diarreia com duração de 7 dias e teste rápido para COVID-19 positivo, evoluiu com disfunção ventricular (FE=56% anterior de 66%) com ressonância magnética compatível com miocardite aguda e elevação de troponina. Após cinco dias de pulsoterapia com corticoide em altas doses e melhora da função ventricular e redução da troponina, realizou biópsia endomicárdica, com resultado 0R. A tomografia computadorizada de coronárias mostrou ausência de calcificação coronária ou redução luminal coronária significativa. A última vacina para COVID foi há 6 meses antes da internação e ela não tomou a terceira dose porque estava gripada na época. Recebeu alta hospitalar após 11 dias com recuperação da função do enxerto. No dia da biópsia, o teste rápido para COVID-19 foi negativo e o RT-PCR no tecido miocárdico não detectou o vírus SARS-CoV-2. Usando a mesma amostra de biópsia, a expressão de 84 genes relacionados à regulação da inflamação também foi analisada usando o método RT2 Profiler PCR Array (Qiagen Cat. No. 330213 PAHS-077Z). A interpretação dos resultados foi comparada com uma amostra controle, que teoricamente não tinha COVID-19. Encontramos 15 genes mais expressos em tecidos que tinham COVID-19 (tabela). Por exemplo, o gene CCL16 foi 8,5 vezes mais expresso em tecido com COVID-19 do que o controle. DISCUSSÃO: A literatura recente descreve que os genes CXCL5 e NFKB1 têm envolvimento na biologia do coronavírus e estão envolvidos na resposta imune ou atividade antiviral, IL17A tem envolvimento na biologia do coronavírus e está envolvido na resposta inflamatória da tempestade de citocinas, e KNG1 tem envolvimento na biologia do coronavírus e é relevante para o processo da doença. CONCLUSÃO: Este é um dos primeiros relatos sobre o uso de um perfil de citocinas analisado em uma biópsia miocárdica de um paciente transplantado com COVID-19. Além da detecção de genes que regulam a inflamação, foram identificados alguns genes relacionados ao envolvimento da biologia e resposta imune ou atividade viral, o que pode ser o primeiro passo para o desenvolvimento de futuros estudos clínicos.

Sequential IgG antibody monitoring for virus-inactivated and adenovirus-vectored COVID-19 vaccine in Brazilian healthcare workers.

Vaccination certainly is the best way to fight against the COVID-19 pandemic. In this study, the seroconversion effectiveness of two vaccines against severe acute respiratory syndrome coronavirus 2 was assessed in healthcare workers: virus-inactivated CoronaVac (CV, n = 303), and adenovirus-vectored Oxford-AstraZeneca (AZ, n = 447). The immunoglobulin G (IgG) antibodies anti-spike glycoprotein and anti-nucleocapsid protein were assessed by enzyme-linked immunosorbent assay at the time before vaccination (T1), before the second dose (T2), and 30 days after the second dose (T3). Of all individuals vaccinated with AZ, 100% (n = 447) exhibited seroconversion, compared to 91% (n = 276) that were given CV vaccine. Among individuals who did not respond to the CV, only three individuals showed a significant increase in the antibody level 4 months later the booster dose. A lower seroconversion rate was observed in elders immunized with the CV vaccine probably due to the natural immune senescence, or peculiarity of this vaccine. The AZ vaccine induced a higher humoral response; however, more common side effects were also observed. Nonvaccinated convalescent individuals revealed a similar rate of anti-spike IgG to individuals that were given two doses of CV vaccine, which suggests that only a one-shot COVID-19 vaccine could produce an effective immune response in convalescents.

Sequential IgG antibody monitoring for virus­inactivated and adenovirus­vectored COVID­19 vaccine in Brazilian health care workers

Vaccination certainly is the best way to fight against the COVID­19 pandemic. In this study, the seroconversion effectiveness of two vaccines against severe acute respiratory syndrome coronavirus 2 was assessed in healthcare workers: virus­inactivated CoronaVac (CV, n= 303), and adenovirus­vectored Oxford­AstraZeneca (AZ, n= 447). The immunoglobulin G (IgG) antibodies anti­spike glycoprotein and anti­nucleocapsid protein were assessed by enzyme­linked immunosorbent assay at the time before vaccination (T1), before the second dose (T2), and 30 days after the second dose (T3). Of all individuals vaccinated with AZ, 100% (n= 447) exhibited seroconversion, compared to 91% (n= 276) that were given CV vaccine. Among individuals who did not respond to the CV, only three individuals showed a significant increase in the antibody level 4 months later the booster dose. A lower seroconversion rate was observed in elders immunized with the CV vaccine probably due to the natural immune senescence, or peculiarity of this vaccine. The AZ vaccine induced a higher humoral response; however, more common side effects were also observed. Nonvaccinated convalescent individuals revealed a similar rate of anti­spike IgG to individuals that were given two doses of CV vaccine, which suggests that only a one­shot COVID­19 vaccine could produce an effective immune response in convalescents.