RESUMO
Tumor necrosis factor (TNF) and interferon-gamma (IFNγ) are important inflammatory mediators in the development of cytokine storm syndrome (CSS). Single nucleotide polymorphisms (SNPs) regulate the expression of these cytokines, making host genetics a key factor in the prognosis of COVID-19. In this study, we investigated the associations of the TNF -308G/A and IFNG +874T/A polymorphisms with COVID-19. We analyzed the frequencies of the two polymorphisms in the control groups (CG: TNF -308G/A, n = 497; IFNG +874T/A, n = 397), a group of patients with COVID-19 (CoV, n = 222) and among the subgroups of patients with nonsevere (n = 150) and severe (n = 72) COVID-19. We found no significant difference between the genotypic and allelic frequencies of TNF -308G/A in the groups analyzed; however, both the frequencies of the high expression genotype (TT) (CoV: 13.51% vs. CG: 6.30%; p = 0.003) and the *T allele (CoV: 33.56% vs. CG: 24. 81%; p = 0.001) of the IFNG +874T/A polymorphism were higher in the COVID-19 group than in the control group, with no differences between the subgroups of patients with nonsevere and severe COVID-19. The *T allele of IFNG +874T/A (rs2430561) is associated with susceptibility to symptomatic COVID-19. These SNPs provided valuables clues about the potential mechanism involved in the susceptibility to developing symptomatic COVID-19.
Assuntos
COVID-19 , Predisposição Genética para Doença , Genótipo , Interferon gama , SARS-CoV-2 , Feminino , Humanos , Masculino , Alelos , COVID-19/genética , COVID-19/virologia , Síndrome da Liberação de Citocina/genética , Frequência do Gene , Interferon gama/genética , Polimorfismo de Nucleotídeo Único , SARS-CoV-2/patogenicidade , Fator de Necrose Tumoral alfa/genéticaRESUMO
Carbapenem-resistant Klebsiella pneumoniae (CRKP) exhibit high mortality rates in pediatric patients and usually belong to international high-risk clones. This study aimed to investigate the molecular epidemiology and carbapenem resistance mechanisms of K. pneumoniae isolates recovered from pediatric patients, and correlate them with phenotypical data. Twenty-five CRKP isolates were identified, and antimicrobial susceptibility was assessed using broth microdilution. Carbapenemase production and ß-lactamase genes were detected by phenotypic and genotypic tests. Multilocus sequence typing was performed to differentiate the strains and whole-genome sequencing was assessed to characterize a new sequence type. Admission to the intensive care unit and the use of catheters were significantly positive correlates of CRKP infection, and the mortality rate was 36%. Almost all isolates showed multidrug-resistant phenotype, and most frequent resistant gene was blaKPC. We observed the dissemination of ST307 and clones belonging to CG258, which are considered high risk. In pediatric patients, these clones present with high genomic plasticity, favoring adaptation of the KPC and NDM enzymes to healthcare environments.
Assuntos
Antibacterianos , Carbapenêmicos , Infecções por Klebsiella , Klebsiella pneumoniae , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , beta-Lactamases , Humanos , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Klebsiella pneumoniae/classificação , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/epidemiologia , Brasil , Criança , Antibacterianos/farmacologia , beta-Lactamases/genética , beta-Lactamases/metabolismo , Carbapenêmicos/farmacologia , Pré-Escolar , Lactente , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/classificação , Masculino , Feminino , Proteínas de Bactérias/genética , Sequenciamento Completo do Genoma , Adolescente , Genótipo , Epidemiologia Molecular , Farmacorresistência Bacteriana Múltipla/genéticaRESUMO
The cGAS-STING pathway appears to contribute to dysregulated inflammation during coronavirus disease 2019 (COVID-19); however, inflammatory factors related to long COVID are still being investigated. In the present study, we evaluated the association of cGAS and STING gene expression levels and plasma IFN-α, TNF-α and IL-6 levels with COVID-19 severity in acute infection and long COVID, based on analysis of blood samples from 148 individuals, 87 with acute COVID-19 and 61 in the post-COVID-19 period. Quantification of gene expression was performed by real-time PCR, and cytokine levels were quantified by ELISA and flow cytometry. In acute COVID-19, cGAS, STING, IFN-α, TNF-α, and IL-6 levels were higher in patients with severe disease than in those with nonsevere manifestations (p < 0.05). Long COVID was associated with elevated cGAS, STING and IFN-α levels (p < 0.05). Activation of the cGAS-STING pathway may contribute to an intense systemic inflammatory state in severe COVID-19 and, after infection resolution, induce an autoinflammatory disease in some tissues, resulting in long COVID.
Assuntos
COVID-19 , Síndrome de COVID-19 Pós-Aguda , Humanos , Interferon-alfa , Interleucina-6 , Nucleotidiltransferases/genética , Nucleotidiltransferases/metabolismo , Transdução de Sinais/genética , Fator de Necrose Tumoral alfa/genéticaRESUMO
In brief: Pyruvate metabolism is one of the main metabolic pathways during oocyte maturation. This study demonstrates that pyruvate metabolism also regulates the epigenetic and molecular maturation in bovine oocytes. Abstract: Pyruvate, the final product of glycolysis, undergoes conversion into acetyl-CoA within the mitochondria of oocytes, serving as a primary fuel source for the tricarboxylic acid (TCA) cycle. The citrate generated in the TCA cycle can be transported to the cytoplasm and converted back into acetyl-CoA. This acetyl-CoA can either fuel lipid synthesis or act as a substrate for histone acetylation. This study aimed to investigate how pyruvate metabolism influences lysine 9 histone 3 acetylation (H3K9ac) dynamics and RNA transcription in bovine oocytes during in vitro maturation (IVM). Bovine cumulus-oocyte complexes were cultured in vitro for 24 h, considering three experimental groups: Control (IVM medium only), DCA (IVM supplemented with sodium dichloroacetate, a stimulant of pyruvate oxidation into acetyl-CoA), or IA (IVM supplemented with sodium iodoacetate, a glycolysis inhibitor). The results revealed significant alterations in oocyte metabolism in both treatments, promoting the utilization of lipids as an energy source. These changes during IVM affected the dynamics of H3K9ac, subsequently influencing the oocyte's transcriptional activity. In the DCA and IA groups, a total of 148 and 356 differentially expressed genes were identified, respectively, compared to the control group. These findings suggest that modifications in pyruvate metabolism trigger the activation of metabolic pathways, particularly lipid metabolism, changing acetyl-CoA availability and H3K9ac levels, ultimately impacting the mRNA content of in vitro matured bovine oocytes.
Assuntos
Histonas , Técnicas de Maturação in Vitro de Oócitos , Animais , Bovinos , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Acetilcoenzima A/metabolismo , Histonas/metabolismo , Oócitos/metabolismo , Ácido Pirúvico/farmacologia , Ácido Pirúvico/metabolismo , Epigênese Genética , Células do CúmuloRESUMO
This study aims to identify the individual community strategies to avoid violence exposure most used by adolescents from public and private schools in the IX Administrative Region of Rio de Janeiro and investigate the profile of co-occurrence and its prevalence in specific population subgroups. This is a cross-sectional study with 693 individuals. A multidimensional questionnaire collected information regarding strategies to avoid community violence exposure and was self-completed in the classroom. The most used strategies were avoiding walking close to armed people (55.5%), avoiding walking alone (30.5%), and avoiding returning home at dawn (24.7%). Girls adopt more of all (concurrently) the four limiting behaviors to reduce their community violence exposure (53% vs. 32%). Notably, the adoption of such strategies differed by socioeconomic indicators and was higher among adolescents from lower-income households. These findings point to the high frequency of use of such strategies by adolescents, which may hinder and limit the full development of their social and cultural skills.
O objetivo do estudo é conhecer as estratégias individuais mais utilizadas por adolescentes de escolas públicas e privadas da IX Região Administrativa do município do Rio de Janeiro para evitar a exposição à violência comunitária, bem como investigar o perfil de coocorrência e sua prevalência em subgrupos populacionais específicos. Trata-se de um estudo seccional com 693 indivíduos. As informações referentes às estratégias para evitar a exposição à violência comunitária foram coletadas por meio de questionário multidimensional autopreenchido em sala de aula. As estratégias mais utilizadas foram: evitar passar onde há pessoas armadas (55,5%), evitar andar sozinho (30,5%) e evitar voltar para casa de madrugada (24,7%). Observou-se que as meninas adotam mais todos (concomitantemente) os quatro tipos de comportamento limitantes para reduzir sua exposição à violência comunitária (53% vs. 32%). Ressalta-se que a adoção de tais estratégias diferiu segundo os indicadores socioeconômicos, sendo maior entre os adolescentes oriundos de família de estratos de renda mais baixos. Tais achados chamam a atenção para a alta frequência de utilização de tais estratégias por adolescentes, o que pode cercear e limitar o pleno desenvolvimento de suas habilidades sociais e culturais.
Assuntos
Exposição à Violência , Feminino , Humanos , Adolescente , Brasil/epidemiologia , Estudos Transversais , Exposição à Violência/prevenção & controle , Renda , Instituições AcadêmicasRESUMO
The oviduct provides a suitable microenvironment from the gametes' final maturation until initial embryo development. Dynamic functional changes are observed in the oviduct cells, mainly controlled by steroid hormones and well-orchestrated during the estrous cycle. However, based on the roles played by the oviduct, additional layers of complexity might be present in its regulatory process. There is a cellular process that includes metabolic adaptation that can guide molecular modifications. This process is known as metaboloepigenetics. Therefore, we aimed to better understand how this crosstalk occurs in oviductal epithelial cells (OEC). Due to limited in situ access to the oviduct, we used the primary in vitro cell culture as a culture model and glucose as a metabolic disturbed factor. For that, cells derived from the oviductal epithelial layer were collected from cows at either follicular or luteal stages (n = 4 animals per group). They were cultured on a monolayer culture system under normoglycemic (2.7 mM glucose) or hyperglycemic conditions (27 mM glucose). On day five of culture, attached cells were submitted to analysis of mitochondrial metabolism (mitochondrial membrane potential - MMP) and epigenetics markers (5- methylcytosine - 5 mC and histone 3 lysine 9 acetylation - H3K9ac). Moreover, the culture media were submitted to the metabolites analysis profile by Raman spectrometry. Data were analyzed considering the effect of glucose level (normoglycemic vs. hyperglycemic), stages when OEC were harvested (follicular vs. luteal), and their interaction (glucose level * cycle stage) by two-way ANOVA. As a result, the high glucose level decreased the H3K9ac and MMP levels but did not affect the 5 mC. Regardless of the metabolic profile of the culture media, the glucose level was the only factor that changed the Raman shifts abundance. Although this present study evaluated oviductal epithelial cells after being submitted to an in vitro monolayer culture system, which is known to lead to cell dedifferentiation, yet, these results provide evidence of a relationship between epigenetic reprogramming and energy metabolism under these cell culture conditions. In conclusion, the levels of metabolites in culture media may be crucial for cellular function and differentiation, meaning that it should be considered in studies culturing oviductal cells.
Assuntos
Tubas Uterinas , Oviductos , Feminino , Animais , Bovinos , Oviductos/metabolismo , Células Epiteliais/metabolismo , Epigênese Genética , Meios de Cultura , Glucose/farmacologia , Glucose/metabolismoRESUMO
OBJECTIVES: We aimed to investigate an outbreak of vancomycin-resistant Enterococcus faecium (VREfm) in paediatric patients from Hospital Pequeno Príncipe. The susceptibility profile was determined, and whole-genome sequencing (WGS) was used to analyse the genetic context of the strains. METHODS: Five VREfm isolates were recovered from sterile sites and surveillance cultures of two paediatric patients with acute lymphoblastic leukaemia. Species identification was performed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and the minimum inhibitory concentration (MIC) was assessed according to the European Committee for Antimicrobial Susceptibility Testing (EUCAST). WGS was performed to analyse the genetic context of virulence and resistance genes, and in silico multilocus sequence typing was performed to identify the sequence typing of the strains. RESULTS: High-level vancomycin resistance was observed in all isolates (≥256 mg/L). WGS revealed the presence of mobile genetic elements, such as plasmids (rep2, rep11a, repUS15, rep17, and rep18a), insertion sequences, and phages. Multiple resistance genes (aac(6')-aph(2"), dfrG, ermB, and vanA) and virulence genes (acm and efaAfm) were identified. All the isolates were assigned to ST117 (ST1133 - via a novel MLST), an important epidemic lineage associated with nosocomial infections and outbreaks. CONCLUSION: Our results show that the ST117 (ST1133) VREfm isolates are circulating in paediatric patients, which raises a great concern. The development of new drugs as well as the implementation of an antimicrobial stewardship program are necessary for their correct management, limiting the spread of resistance in oncohematological patients.
Assuntos
Enterococcus faecium , Leucemia-Linfoma Linfoblástico de Células Precursoras , Enterococos Resistentes à Vancomicina , Humanos , Criança , Vancomicina/farmacologia , Tipagem de Sequências Multilocus , Brasil/epidemiologia , Genótipo , Enterococos Resistentes à Vancomicina/genética , Surtos de DoençasRESUMO
Resumo A pandemia de Covid-19 produziu uma série de agravos sociais, como crescimento exponencial da pobreza e da fome, aumento nas taxas de desemprego, interrupção de atividades de trabalhadores informais, piora das condições de moradia da população de baixa renda e transtornos psíquicos. No Brasil, essa realidade foi enfrentada de forma insuficiente no que se refere às ações minimizadoras do governo, incluindo programas de transferência de renda. Ainda assim, apenas no primeiro ano de pandemia, a concessão dos chamados benefícios eventuais passou de 3 para 7,8 milhões de pessoas. Apesar da existência de muitos estudos estatísticos sobre a piora das condições socioeconômicas da população, ainda são raros os estudos que investigam o problema na perspectiva das pessoas socioeconomicamente mais atingidas. A pesquisa ouviu 27 chefes de família de baixa renda, atendidos em um CRAS do DF, sobre os efeitos da pandemia em suas vidas. Os resultados permitiram compreender, em profundidade, os sofrimentos e dificuldades vividas em aspectos como: trabalho e renda; alimentação; educação de crianças; sofrimentos psíquicos; obtenção e uso de benefícios eventuais e apontam para novas estratégias, programas e políticas capazes de mitigar os efeitos da pandemia que perdurarão ainda por longo tempo.
Abstract The Covid-19 pandemic produced a series of social problems, such as the exponential growth of poverty and hunger, an increase in unemployment rates, interruption of activities of informal workers, worsening of the housing conditions of the low-income population and psychological disorders. In Brazil, this reality has been insufficiently addressed in terms of minimizing government actions, including income transfer programs. Even so, only in the first year of the pandemic, the granting of so-called occasional benefits went from 3 to 7.8 million people. Despite the existence of many statistical studies on the worsening of the socioeconomic conditions of the population, studies that investigate the problem from the perspective of the most socioeconomically affected people are still rare. The research heard 27 low-income family heads, assisted at a CRAS in the DF, Brazil, about the effects of the pandemic on their daily lives. The results made it possible to understand, in depth, the sufferings and difficulties experienced in aspects such as: work and income; food; children's education; psychic sufferings; obtaining and using eventual benefits and point to new strategies, programs and policies capable of mitigating the effects of the pandemic that will last for a long time.Resumo: A pandemia de Covid-19 produziu uma série de agravos sociais, como crescimento exponencial da pobreza e da fome, aumento nas taxas de desemprego, interrupção de atividades de trabalhadores informais, piora das condições de moradia da população de baixa renda e transtornos psíquicos. No Brasil, essa realidade foi enfrentada de forma insuficiente no que se refere às ações minimizadoras do governo, incluindo programas de transferência de renda. Ainda assim, apenas no primeiro ano de pandemia, a concessão dos chamados benefícios eventuais passou de 3 para 7,8 milhões de pessoas. Apesar da existência de muitos estudos estatísticos sobre a piora das condições socioeconômicas da população, ainda são raros os estudos que investigam o problema na perspectiva das pessoas socioeconomicamente mais atingidas. A pesquisa ouviu 27 chefes de família de baixa renda, atendidos em um CRAS do DF, sobre os efeitos da pandemia em suas vidas. Os resultados permitiram compreender, em profundidade, os sofrimentos e dificuldades vividas em aspectos como: trabalho e renda; alimentação; educação de crianças; sofrimentos psíquicos; obtenção e uso de benefícios eventuais e apontam para novas estratégias, programas e políticas capazes de mitigar os efeitos da pandemia que perdurarão ainda por longo tempo.
RESUMO
Resumo O objetivo do estudo é conhecer as estratégias individuais mais utilizadas por adolescentes de escolas públicas e privadas da IX Região Administrativa do município do Rio de Janeiro para evitar a exposição à violência comunitária, bem como investigar o perfil de coocorrência e sua prevalência em subgrupos populacionais específicos. Trata-se de um estudo seccional com 693 indivíduos. As informações referentes às estratégias para evitar a exposição à violência comunitária foram coletadas por meio de questionário multidimensional autopreenchido em sala de aula. As estratégias mais utilizadas foram: evitar passar onde há pessoas armadas (55,5%), evitar andar sozinho (30,5%) e evitar voltar para casa de madrugada (24,7%). Observou-se que as meninas adotam mais todos (concomitantemente) os quatro tipos de comportamento limitantes para reduzir sua exposição à violência comunitária (53% vs. 32%). Ressalta-se que a adoção de tais estratégias diferiu segundo os indicadores socioeconômicos, sendo maior entre os adolescentes oriundos de família de estratos de renda mais baixos. Tais achados chamam a atenção para a alta frequência de utilização de tais estratégias por adolescentes, o que pode cercear e limitar o pleno desenvolvimento de suas habilidades sociais e culturais.
Abstract This study aims to identify the individual community strategies to avoid violence exposure most used by adolescents from public and private schools in the IX Administrative Region of Rio de Janeiro and investigate the profile of co-occurrence and its prevalence in specific population subgroups. This is a cross-sectional study with 693 individuals. A multidimensional questionnaire collected information regarding strategies to avoid community violence exposure and was self-completed in the classroom. The most used strategies were avoiding walking close to armed people (55.5%), avoiding walking alone (30.5%), and avoiding returning home at dawn (24.7%). Girls adopt more of all (concurrently) the four limiting behaviors to reduce their community violence exposure (53% vs. 32%). Notably, the adoption of such strategies differed by socioeconomic indicators and was higher among adolescents from lower-income households. These findings point to the high frequency of use of such strategies by adolescents, which may hinder and limit the full development of their social and cultural skills.
RESUMO
The epigenetic reprogramming that occurs during the earliest stages of embryonic development has been described as crucial for the initial events of cell specification and differentiation. Recently, the metabolic status of the embryo has gained attention as one of the main factors coordinating epigenetic events. In this work, we investigate the link between pyruvate metabolism and epigenetic regulation by culturing bovine embryos from day 5 in the presence of dichloroacetate (DCA), a pyruvate analog that increases the pyruvate to acetyl-CoA conversion, and iodoacetate (IA), which inhibits the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), leading to glycolysis inhibition. After 8 h of incubation, both DCA and IA-derived embryos presented higher mitochondrial membrane potential. Nevertheless, in both cases, lower levels of acetyl-CoA, ATP-citrate lyase and mitochondrial membrane potential were found in blastocysts, suggesting an adaptative metabolic response, especially in the DCA group. The metabolic alteration found in blastocysts led to changes in the global pattern of H3K9 and H3K27 acetylation and H3K27 trimethylation. Transcriptome analysis revealed that such alterations resulted in molecular differences mainly associated to metabolic processes, establishment of epigenetic marks, control of gene expression and cell cycle. The latter was further confirmed by the alteration of total cell number and cell differentiation in both groups when compared to the control. These results corroborate previous evidence of the relationship between the energy metabolism and the epigenetic reprogramming in preimplantation bovine embryos, reinforcing that the culture system is decisive for precise epigenetic reprogramming, with consequences for the molecular control and differentiation of cells.
Assuntos
Epigênese Genética , Transcriptoma , Feminino , Gravidez , Animais , Bovinos , Acetilcoenzima A/metabolismo , Desenvolvimento Embrionário/genética , Blastocisto/metabolismo , Perfilação da Expressão Gênica , Piruvatos/metabolismoRESUMO
Interleukin-6 has been recognized as a major role player in COVID-19 severity, being an important regulator of the cytokine storm. Hence, the evaluation of the influence of polymorphisms in key genes of the IL-6 pathway, namely IL6, IL6R, and IL6ST, may provide valuable prognostic/predictive markers for COVID-19. The present cross-sectional study genotyped three SNPs (rs1800795, rs2228145, and rs7730934) at IL6. IL6R and IL6ST genes, respectively, in 227 COVID-19 patients (132 hospitalized and 95 non-hospitalized). Genotype frequencies were compared between these groups. As a control group, published data on gene and genotype frequencies were gathered from published studies before the pandemic started. Our major results point to an association of the IL6 C allele with COVID-19 severity. Moreover, IL-6 plasmatic levels were higher among IL6 CC genotype carriers. Additionally, the frequency of symptoms was higher at IL6 CC and IL6R CC genotypes. In conclusion, the data suggest an important role of IL6 C allele and IL6R CC genotype on COVID-19 severity, in agreement with indirect evidence from the literature about the association of these genotypes with mortality rates, pneumonia, and heightening of protein plasmatic levels pro-inflammatory driven effects.
Assuntos
COVID-19 , Interleucina-6 , Humanos , Interleucina-6/genética , Estudos Transversais , Receptores de Interleucina-6/genética , COVID-19/genética , Genótipo , Polimorfismo de Nucleotídeo Único , Predisposição Genética para Doença , Receptor gp130 de Citocina/genéticaRESUMO
Introduction: Mannose-binding lectin (MBL) promotes opsonization, favoring phagocytosis and activation of the complement system in response to different microorganisms, and may influence the synthesis of inflammatory cytokines. This study investigated the association of MBL2 gene polymorphisms with the plasma levels of MBL and inflammatory cytokines in COVID-19. Methods: Blood samples from 385 individuals (208 with acute COVID-19 and 117 post-COVID-19) were subjected to real-time PCR genotyping. Plasma measurements of MBL and cytokines were performed by enzyme-linked immunosorbent assay and flow cytometry, respectively. Results: The frequencies of the polymorphic MBL2 genotype (OO) and allele (O) were higher in patients with severe COVID-19 (p< 0.05). The polymorphic genotypes (AO and OO) were associated with lower MBL levels (p< 0.05). IL-6 and TNF-α were higher in patients with low MBL and severe COVID-19 (p< 0.05). No association of polymorphisms, MBL levels, or cytokine levels with long COVID was observed. Discussion: The results suggest that, besides MBL2 polymorphisms promoting a reduction in MBL levels and therefore in its function, they may also contribute to the development of a more intense inflammatory process responsible for the severity of COVID-19.
Assuntos
COVID-19 , Lectina de Ligação a Manose , Humanos , Fator de Necrose Tumoral alfa/genética , Interleucina-6/genética , Citocinas/genética , Síndrome de COVID-19 Pós-Aguda , COVID-19/genética , Polimorfismo Genético , Lectina de Ligação a Manose/genéticaRESUMO
The first case of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in Brazil was diagnosed on February 26, 2020. Due to the important epidemiological impact of COVID-19, the present study aimed to analyze the specificity of IgG antibody responses to the S1, S2 and N proteins of SARS-CoV-2 in different COVID-19 clinical profiles. This study enrolled 136 individuals who were diagnosed with or without COVID-19 based on clinical findings and laboratory results and classified as asymptomatic or as having mild, moderate or severe disease. Data collection was performed through a semistructured questionnaire to obtain demographic information and main clinical manifestations. IgG antibody responses to the S1 and S2 subunits of the spike (S) protein and the nucleocapsid (N) protein were evaluated using an enzyme-linked immunosorbent assay (ELISA) according to the manufacturer's instructions. The results showed that among the participants, 87.5% (119/136) exhibited IgG responses to the S1 subunit and 88.25% (120/136) to N. Conversely, only 14.44% of the subjects (21/136) displayed S2 subunit responses. When analyzing the IgG antibody response while considering the different proteins of the virus, patients with severe disease had significantly higher antibody responses to N and S1 than asymptomatic individuals (p ≤ 0.0001), whereas most of the participants had low antibody titers against the S2 subunit. In addition, individuals with long COVID-19 showed a greater IgG response profile than those with symptomatology of a short duration. Based on the results of this study, it is concluded that levels of IgG antibodies may be related to the clinical evolution of COVID-19, with high levels of IgG antibodies against S1 and N in severe cases and in individuals with long COVID-19.
Assuntos
COVID-19 , Humanos , Anticorpos Antivirais , Formação de Anticorpos , Imunoglobulina G , Proteínas do Nucleocapsídeo , Síndrome de COVID-19 Pós-Aguda , SARS-CoV-2 , Glicoproteína da Espícula de CoronavírusRESUMO
Lipids play a crucial role in various biological functions, including membrane composition, energy storage, cell signalling, and metabolic and epigenetic processes. Abnormal lipid accumulation and metabolism during in vitro maturation (IVM) of oocytes have been linked to the use of fetal bovine serum (FBS), even though it provides several beneficial molecules, contributing to the oocyte competence. Delipidating agents have been used to mitigate these deleterious effects, but they can have adverse effects on embryonic development. In this study, we explored how lipids present in fetal bovine serum (FBS) can impact the composition of oocytes and their resulting blastocysts in vitro. For that, we used organic solvents to separate the polar and nonpolar (lipid enriched) phase of FBS. Oocytes were in vitro matured in the presence of 10% whole FBS (control), 10% FBS plus 10% nonpolar lipids (lipid enriched - OL) or 10% polar lipids only (partially delipidated - ODL). After 24 h, part of the matured oocytes was collected and those remaining in each group underwent in vitro fertilization (IVF) and culture (IVC) under the same conditions and expanded blastocysts were collected at day 7 (control, BL and BDL). Oocytes and embryos were analysed by Multiple Reaction Monitoring mass spectrometry (MRM-MS) to determine their lipid composition. Interestingly, principal component analysis (PCA) revealed a clear distinction in the lipid profile of oocytes and blastocysts from both treatments compared to the control group. Control oocytes and blastocysts had higher triacylglycerol and cholesterol ester enrichment while the OL, ODL, BL and BDL groups had higher amounts of free fatty acids (FFAs). The structural and signalling phospholipids also differed among groups. Our findings suggest that the lipid-enriched fraction of FBS can be manipulated for IVM to ensure proper maturation, resulting in oocytes and blastocysts with less accumulated intracellular lipids and an improved metabolic status.
Assuntos
Oócitos , Soroalbumina Bovina , Gravidez , Feminino , Animais , Oócitos/metabolismo , Desenvolvimento Embrionário , Fertilização in vitro/veterinária , Blastocisto/metabolismo , Triglicerídeos/metabolismo , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodosRESUMO
Supplementation of culture media with IGF-1 during in vitro culture of embryos has had controversial results over the years. In the present study, we show that differences previously observed in response to IGF addition might be related to intrinsic heterogeneity of the embryos. In other words, the effects exerted by IGF-1 are dependent on the characteristics of the embryos and their ability to modulate metabolism and overcome stressful conditions, such as the ones found in a non-optimized in vitro culture system. To test this hypothesis, in vitro produced bovine embryos with distinct morphokinetics (fast- and slow-cleavage) were submitted to treatment with IGF-1 and then evaluated for embryo production rates, total cell number, gene expression and lipid profile. Our results show that remarkable differences were found when fast and slow embryos treated with IGF-1 were compared. Fast embryos respond by upregulating genes related to mitochondrial function, stress response, and lipid metabolism, whereas slow embryos presented lower mitochondrial efficiency and lipid accumulation. We conclude that indeed the treatment with IGF-1 selectively affects embryonic metabolism according to early morphokinetics phenotypes, and this information is relevant for decision-making in the design of more appropriate in vitro culture systems.
Assuntos
Desenvolvimento Embrionário , Fator de Crescimento Insulin-Like I , Animais , Bovinos , Fator de Crescimento Insulin-Like I/metabolismo , Desenvolvimento Embrionário/fisiologia , Blastocisto/fisiologia , Embrião de Mamíferos , Lipídeos , Fertilização in vitro/métodos , Fertilização in vitro/veterináriaRESUMO
Aiming to evaluate the role of ten functional polymorphisms in long COVID, involved in major inflammatory, immune response and thrombophilia pathways, a cross-sectional sample composed of 199 long COVID (LC) patients and a cohort composed of 79 COVID-19 patients whose follow-up by over six months did not reveal any evidence of long COVID (NLC) were investigated to detect genetic susceptibility to long COVID. Ten functional polymorphisms located in thrombophilia-related and immune response genes were genotyped by real time PCR. In terms of clinical outcomes, LC patients presented higher prevalence of heart disease as preexistent comorbidity. In general, the proportions of symptoms in acute phase of the disease were higher among LC patients. The genotype AA of the interferon gamma (IFNG) gene was observed in higher frequency among LC patients (60%; p = 0.033). Moreover, the genotype CC of the methylenetetrahydrofolate reductase (MTHFR) gene was also more frequent among LC patients (49%; p = 0.045). Additionally, the frequencies of LC symptoms were higher among carriers of IFNG genotypes AA than among non-AA genotypes (Z = 5.08; p < 0.0001). Two polymorphisms were associated with LC in both inflammatory and thrombophilia pathways, thus reinforcing their role in LC. The higher frequencies of acute phase symptoms among LC and higher frequency of underlying comorbidities might suggest that acute disease severity and the triggering of preexisting condition may play a role in LC development.
Assuntos
COVID-19 , Trombofilia , Humanos , Síndrome de COVID-19 Pós-Aguda , Frequência do Gene , Marcadores Genéticos , Estudos Transversais , COVID-19/genética , Genótipo , Predisposição Genética para Doença , Trombofilia/genética , Polimorfismo de Nucleotídeo Único , Estudos de Casos e ControlesRESUMO
The laboratory diagnosis of Clostridioides difficile infection (CDI) is challenging since this bacteria may be detected in healthy people and toxin production detection is not sensitive enough to be used alone. Thus, there is no single test with adequate sensitivity and specificity to be used in laboratory diagnosis. We evaluated the performance of tests used in the diagnosis of CDI in symptomatic patients with risk factors in hospitals in southern Brazil. Enzyme immunoassays (EIA) for glutamate dehydrogenase antigen (GDH) and toxins A/B, real-time polymerase chain reaction (qPCR), GeneXpert system, and a two-step algorithm comprising GDH/TOXIN EIA performed simultaneously followed by GeneXpert for outliers were evaluated. Toxigenic strain in stool culture was considered CDI positive (gold standard). Among 400 samples tested, 54 (13.5%) were positive for CDI and 346 (86.5%) were negative. The diagnosis of the two-step algorithm and qPCR had an excellent performance with an accuracy of 94.5% and 94.2%, respectively. The Youden index showed that GeneXpert as a single test (83.5%) and the two-step algorithm (82.8%) were the most effective assays. Diagnosing CDI and non-CDI diarrhea could be successfully attained by the combination of clinical data with accuracy of laboratory tests.
Assuntos
Toxinas Bacterianas , Clostridioides difficile , Infecções por Clostridium , Humanos , Toxinas Bacterianas/genética , Toxinas Bacterianas/análise , Clostridioides difficile/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/análise , Fezes/microbiologia , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/microbiologia , Enterotoxinas , Sensibilidade e Especificidade , Reação em Cadeia da Polimerase em Tempo Real , Glutamato Desidrogenase/análise , Técnicas de Laboratório ClínicoRESUMO
BACKGROUND: Fanconi-Debré-de Toni syndrome (also known as Fanconi renotubular syndrome, or FRST) profoundly increased the understanding of the functions of the proximal convoluted tubule (PCT) and provided important insights into the pathophysiology of several kidney diseases and drug toxicities. DATA SOURCES: We searched Pubmed and Scopus databases to find relevant articles about FRST. This review article focuses on the physiology of the PCT, as well as on the physiopathology of FRST in children, its diagnosis, and treatment. RESULTS: FRST encompasses a wide variety of inherited and acquired PCT alterations that lead to impairment of PCT reabsorption. In children, FRST often presents as a secondary feature of systemic disorders that impair energy supply, such as Lowe's syndrome, Dent's disease, cystinosis, hereditary fructose intolerance, galactosemia, tyrosinemia, Alport syndrome, and Wilson's disease. Although rare, congenital causes of FRST greatly impact the morbidity and mortality of patients and impose diagnostic challenges. Furthermore, its treatment is diverse and considers the ability of the clinician to identify the correct etiology of the disease. CONCLUSION: The early diagnosis and treatment of pediatric patients with FRST improve the prognosis and the quality of life.
Assuntos
Cistinose , Síndrome de Fanconi , Nefropatias , Síndrome Oculocerebrorrenal , Humanos , Criança , Síndrome de Fanconi/diagnóstico , Síndrome de Fanconi/genética , Síndrome de Fanconi/terapia , Qualidade de Vida , Cistinose/complicações , Síndrome Oculocerebrorrenal/complicaçõesRESUMO
The duration and severity of COVID-19 are related to age, comorbidities, and cytokine synthesis. This study evaluated the impact of these factors on patients with clinical presentations of COVID-19 in a Brazilian cohort. A total of 317 patients diagnosed with COVID-19 were included; cases were distributed according to clinical status as severe (n=91), moderate (n=56) and mild (n=170). Of these patients, 92 had acute COVID-19 at sample collection, 90 had already recovered from COVID-19 without sequelae, and 135 had sequelae (long COVID syndrome). In the acute COVID-19 group, patients with the severe form had higher IL-6 levels (p=0.0260). In the post-COVID-19 group, there was no significant difference in cytokine levels between groups with different clinical conditions. In the acute COVID-19 group, younger patients had higher levels of TNF-α, and patients without comorbidities had higher levels of TNF-α, IL-4 and IL-2 (p<0.05). In contrast, patients over age 60 with comorbidities had higher levels of IL-6. In the post-COVID-19 group, subjects with long COVID-19 had higher levels of IL-17 and IL-2 (p<0.05), and subjects without sequelae had higher levels of IL-10, IL-6 and IL- 4 (p<0.05). Our results suggest that advanced age, comorbidities and elevated serum IL-6 levels are associated with severe COVID-19 and are good markers to differentiate severe from mild cases. Furthermore, high serum levels of IL-17 and IL-2 and low levels of IL-4 and IL-10 appear to constitute a cytokine profile of long COVID-19, and these markers are potential targets for COVID-19 treatment and prevention strategies.
Assuntos
Tratamento Farmacológico da COVID-19 , COVID-19 , Biomarcadores , COVID-19/complicações , Citocinas , Humanos , Interleucina-10 , Interleucina-17 , Interleucina-2 , Interleucina-4 , Interleucina-6 , Pessoa de Meia-Idade , SARS-CoV-2 , Fator de Necrose Tumoral alfa , Síndrome de COVID-19 Pós-AgudaRESUMO
The search for gold-standard materials for bone regeneration is still a challenge in reconstruction surgery. The ratio between hydroxyapatite (HAp) and ß-tricalcium phosphate (ß-TCP) in biphasic calcium phosphate ceramics (BCPs) is one of the most important factors in osteoinduction promotion and controlled biodegradability, configurating what is currently considered as a possible gold standard material for bone substitution in reconstructive surgery. Exploring the natural genesis of the HAp and ß-TCP phases in fishbones during their postnatal growth, this study developed a biphasic bioceramic obtained from the calcination of Nile tilapia (Oreochromis niloticus) bones as a function of their ages. The natural genesis dynamics of the structural evolution of the ß-TCP and HAp phases were characterized by physicochemical methods, taking into account of the age of the fish and the material processing conditions. Thermal analysis (TGA / DTA) showed complete removal of the organic matter and transitions associated with the transformation of carbonated hydroxyapatite (CDHA) to HAp and ß-TCP phases. After calcination at 900 °C, the material was characterized by: X-ray diffraction (XRD) and refinement by the Rietveld method; Fourier Transform Infrared Spectroscopy with Attenuated Total Reflection (FTIR-ATR); Raman spectroscopy; Scanning Electron Microscopy (SEM) and Flame Atomic Absorption Spectroscopy (FAAS). The analysis allowed identification and quantitative estimate of the variations of the HAp and ß-TCP phases in the formation of the BCPs. The results showed that the decrease in ß-TCP against the increase in the HAp phases is symmetrical to the dynamics of the natural genesis of these phases, surprisingly maintaining the balanced phase proportion even when bones of young fishes were used. The microstructure analysis confirms the observed transformation. In addition, in vivo tests demonstrated the osteoinductive potential of BCP scaffolds implanted in an ectopic site, and their remarkable regenerative functionality, as bone graft, was demonstrated in alveolar bone after tooth extraction. MTT cytotoxicity assay for BCP samples for MC3T3-E1 pre-osteoblasts and L929 fibroblasts cells showed viability equal or higher than 100%. A logistic empirical model is presented to explain the three stages of HAp natural formation with fish age and it is also compared to the fish size evolution.