RESUMO
In brief: Ubiquitination plays a pivotal role in a multitude of cellular functions; however, the precise contributions of various ubiquitin ligases in governing early developmental processes remain largely unexplored. This study revealed that the E3 ubiquitin ligases DCAF13 and RNF114 are both necessary for the normal regulation of early porcine embryo development. Abstract: Ubiquitylation is required for normal regulation of many biological functions by modulating several protein facets such as structure, stability, interaction, localization, and degradation. In this study, we explored the roles of two E3 ubiquitin ligases (E3s), the DDB1- and CUL4-associated factor 13 (DCAF13) and the Ring finger protein 114 (RNF114), in the regulation of porcine embryo development. Attenuation of DCAF13 mRNA decreased embryo development at the blastocyst stage, while the development of RNF114-attenuated embryos was not significantly different than that of control embryos. The average number of cells per blastocyst was decreased in DCAF13-attenuated embryos and increased in RNF114-attenuated embryos compared to controls. The relative mRNA abundance of the histone methyltransferase SUV39H1, which regulates histone H3 lysine 9 trimethylation (H3K9me3), was increased in both DCAF13- and RNF114-attenuated embryos, but nuclear immunofluorescence signal for H3K9me3 on day 3 embryos was not significantly altered between attenuated and control embryos. Nuclear immunofluorescence signal for H3K4m3 was decreased in DCAF13-attenuated embryos, but it was increased in RNF114-attenuated embryos compared to controls. Attenuation of DCAF13 and RNF114 mRNAs increased transcript levels for the DNA recombinase RAD51 and decreased expression of phosphorylated histone H2A.X (γH2AX), which suggests an impact on DNA damage repair. In addition, lower mRNA expression of the lysine demethylases 5B (KDM5B) and 5C (KDM5C), both involved in embryo genome activation and DNA repair, was detected in DCAF13-attenuated embryos. These findings indicated that both DCAF13 and RNF114 have important roles in the regulation of the early development of porcine embryos.
Assuntos
Desenvolvimento Embrionário , Fator XIII , Suínos , Ubiquitina-Proteína Ligases , Animais , Blastocisto , Desenvolvimento Embrionário/genética , Fator XIII/metabolismo , Lisina/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Suínos/embriologia , Proteínas de Ligação a RNA , Ubiquitina-Proteína Ligases/metabolismoRESUMO
INTRODUCCIÓN. La hemofilia es una condición rara hereditaria, crónica, potencialmente discapacitante e incapacitante, caracterizada por frecuentes sangrados debidos al déficit del factor VIII coagulante, Hemofilia A o del factor IX Hemofilia B. Las evaluaciones de calidad de vida en personas con hemofilia, basadas principalmente en el aspecto biológico, llevaron a considerar un importante enfoque bioético que evalúe la afectación de la autonomía y dignidad debida a la enfermedad. OBJETIVO. Registrar la percepción de la autonomía y dignidad de personas que viven con hemofilia. MATERIALES Y MÉTODOS. Estudio descriptivo transversal. Población de 92 y muestra de 28 varones mayores de 18 años con diagnóstico de hemofilia, atendidos en la Clínica de Coagulopatías Congénitas del Hospital de Especialidades Carlos Andrade Marín en el periodo marzo 2021 a agosto del 2021. Se excluyó a varones menores de 18 años atendidos en otras instituciones del Sistema Nacional de Salud. Estudio basado en el desarrollo de las capacidades centrales descritas por Martha Nussbaum. Se aplicó el test The Hemophilia Well Being Index que evaluó calidad de vida con relación al bienestar personal asociado a salud, y la herramienta Body Mapping que analizó en base al interpretativismo fenomenológico. RESULTADOS. El 100% de personas presentaron afectación en algún área de la vida investigada por el Hemophilia Well Being Index, que se confirma con las expresiones escritas y gráficas recopiladas por el Body Mapping. CONCLUSIÓN. La autonomía y dignidad se encuentran afectadas en las personas que viven con hemofilia, al igual que las capacidades centrales; es importante valorar cómo estos parámetros afectan la consecución de logros, lo que se debe considerar en estudios futuros.
INTRODUCTION. Hemophilia is a rare hereditary, chronic, potentially disabling and incapacitating condition, characterized by frequent bleeds due to deficiency of clotting factor VIII, Hemophilia A or factor IX Hemophilia B. Quality of life assessments in people with hemophilia, mainly based on the biological aspect, led to consider an important bioethical approach that evaluates the impairment of autonomy and dignity due to the disease. OBJECTIVE. To record the perception of autonomy and dignity of people living with hemophilia. MATERIALS AND METHODS. Cross-sectional descriptive study. Population of 92 and sample of 28 males over 18 years of age with a diagnosis of hemophilia, attended at the Congenital Coagulopathy Clinic of the Carlos Andrade Marin Specialty Hospital in the period March 2021 to August 2021. Males under 18 years of age attended in other institutions of the National Health System were excluded. The study was based on the development of the central capabilities described by Martha Nussbaum. The test The Hemophilia Well Being Index was applied, which evaluated quality of life in relation to personal wellbeing associated with health, and the tool Body Mapping which analyzed based on phenomenological interpretivism. RESULTS. 100% of people presented affectation in some area of life investigated by the Hemophilia Well Being Index, which is confirmed by the written and graphic expressions collected by the Body Mapping. CONCLUSION. Autonomy and dignity are affected in people living with hemophilia, as are core capacities; it is important to assess how these parameters affect achievement, which should be considered in future studies.
Assuntos
Humanos , Masculino , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Adulto Jovem , Percepção , Qualidade de Vida , Hemofilia B , Autonomia Pessoal , Assistência ao Paciente , Hemofilia A , Coagulação Sanguínea , Fatores de Coagulação Sanguínea , Fator IX , Fator XIII , Doença Crônica , Direitos Civis , Indicadores de Doenças CrônicasRESUMO
INTRODUCTION: Factor XIII (FXIII) deficiency may cause bleeding under certain clinical circumstances. Therapeutic plasma exchange (TPE) may lead to a transient deficiency. OBJECTIVES: To describe the clinical evolution of patients with acquired FXIII deficiency secondary to TPE. METHODS: We respectively studied a cohort of consecutive patients from 2014 to 2019 who were treated with TPE with FXIII levels <50%. The FXIII was measured after the start of the TPE course, on days between the TPE sessions, due to suspected acquired deficiency. All TPE were performed using continuous flow cell separator. In all cases, the initial replacement fluid applied was albumin. Apheresis procedures were held at 24to 48 hours intervals. RESULTS: Eighteen patients were included, 13 of them were recipients of kidney transplants. The main TPE prescription was humoral rejection. Median FXIII at diagnosis (measured on days between sessions of the TPE course) was 19%(IQR17-25). The median of apheresis procedures before measurement of FXIII was 3(IQR2-4). Among the total cohort, 10 patients suffered hemorrhages. None of the patients without history of kidney transplants had bleeding (n = 5), however, 10/13 with kidney transplants did. Five kidney transplant patients received therapy with FXIII concentrate because of life-threatening bleeding. In all cases, the bleeding stopped within the first 24 hours. All patients had their FXIII levels measured again after finishing the TPE course, with normal results. CONCLUSIONS: TPE is an under-diagnosed cause of acquired FXIII deficiency since routine coagulation tests remain unaltered. It might cause major bleeding, particularly in patients with a recent history of surgery like kidney transplants.
Assuntos
Deficiência do Fator XIII/etiologia , Troca Plasmática/efeitos adversos , Adulto , Fator XIII/análise , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto JovemRESUMO
El Instituto de Evaluación de Tecnologías en Salud e Investigación (IETSI) de EsSalud planteó por lo tanto la realización de una guía de práctica clínica (GPC) peruana para el manejo de hemofilia.
Assuntos
Humanos , Masculino , Hemofilia B/diagnóstico , Hemofilia B/tratamento farmacológico , Hemofilia A/diagnóstico , Hemofilia A/tratamento farmacológico , Índice de Gravidade de Doença , Fator IX/administração & dosagem , Fator XIII/administração & dosagemRESUMO
The evaluation of patients with a bleeding tendency represents a challenge as the routinely available tests for evaluating bleeding disorders are limited, complicating the laboratory determination of the clinically observed bleeding tendency. As a result, some bleeding disorders remain undiagnosed. The aim of the study was to evaluate whether global coagulation tests would contribute to the laboratory analysis of patients with undiagnosed bleeding disorders. Patients were evaluated for coagulation and fibrinolysis activities by thrombin generation test and euglobulin lysis time. In addition, plasma activity of factor XIII, plasminogen, α-2 antiplasmin, plasminogen activator inhibitor-1, and thrombin-activatable fibrinolysis inhibitor was also obtained. Forty-five patients were included. Eight per cent presented a mild bleeding disorder and 20% a moderate bleeding disorder. The thrombin generation test results were similar between patients and controls. Euglobulin lysis time results, however, were lower in patients than in controls, both before (median 175 vs. 250âmin, respectively; Pâ=â0.003) and after (median 145 vs. 115âmin, respectively; Pâ≤â0.001) arm constriction, suggesting that they were experiencing hyperfibrinolysis. Interestingly, patients' median thrombin-activatable fibrinolysis inhibitor activity was higher than in controls (21.2 vs. 19.46âµg/ml; Pâ=â0.016). However, plasminogen, α-2 antiplasmin, plasminogen activator inhibitor-1, and factor XIII activities did not differ between the groups. Global coagulation and fibrinolysis tests proved to be limited in detecting the hemostatic disorders in some patients with a relevant bleeding tendency and may not be adequate to address their bleeding risk. Bleeding scores are currently the available medical approach for the evaluation of these patients.
Assuntos
Coagulação Sanguínea , Transtornos Hemorrágicos/diagnóstico , Projetos de Pesquisa , Adolescente , Adulto , Estudos de Casos e Controles , Fator XIII/metabolismo , Feminino , Tempo de Lise do Coágulo de Fibrina , Transtornos Hemorrágicos/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Plasminogênio/metabolismo , Inibidor 1 de Ativador de Plasminogênio/sangue , Trombina/metabolismo , Ativador de Plasminogênio Tecidual/sangue , alfa 2-Antiplasmina/metabolismoRESUMO
Nijmegen-Bethesda assay is the gold standard to assess inhibitory antibodies against factor (F) VIII. This method has some limitations, including high coefficient of variation and possible interference of residual endogenous or exogenous factor VIII. Heat-treatment of samples at 56 °C for 30 min could be a strategy to improve the sensitivity of this test. The aim of this study was to compare inhibitor quantification in hemophilia patients with and without inhibitor performed in previously heated and non-heated samples. A total of 109 analyses from 46 patients with severe hemophilia A were performed. Patients were divided into three groups: 20 patients with no history of inhibitor, recently and not recently exposed to FVIII (group I), 21 patients with history of inhibitor not exposed to FVIII (group II), and 5 patients (68 samples) undergoing an immune tolerance induction (ITI) protocol (group III). For patients with no history of inhibitor, heat-treatment did not modify the results (p=0.24). However, differences in inhibitor levels between heated and non-heated samples were observed in patients with history of inhibitor (group II, p<0.05) and in patients in ITI (group III, p<0.001). In 11 samples, inhibitor quantification shifted from negative to positive. Additionally, a longitudinal evaluation of each ITI patient showed similar trend line for the results of heated and non-heated samples. In this study, we demonstrated that heating samples increase sensitivity of Nijmegen-Bethesda assay, with no shift from negative to positive results in patients with no history of inhibitor. Furthermore, this procedure has an important role to patients undergoing an ITI protocol.
Assuntos
Fator XIII/química , Testes Hematológicos/métodos , Hemofilia A/imunologia , Autoanticorpos/imunologia , Brasil , Fator VIII/imunologia , Reações Falso-Positivas , Hemofilia A/sangue , Temperatura Alta , Humanos , Sistema Imunitário , Tolerância ImunológicaRESUMO
PURPOSE: To investigate hemostatic effects of supplementary factor XIII and desmopressin (DDAVP) in resuscitation of uncontrolled bleeding. METHODS: Fifty-four rabbits were randomized in nine groups: G1: Sham; G2: FXIII and normotensive resuscitation (NBP); G3: FXIII and permissive hypotension (PH) (MAP 60% baseline); G4: FXIII/DDAVP/NBP; G5: FXIII/DDAVP/PH; G6: NBP only; G7: FXIII no hemorrhage; G8: FXIII/DDAVP no hemorrhage; G9: PH only. Thromboelastometry and intra-abdominal blood loss were assessed. Scanning electron microscopy (EM) of the clots was performed. RESULTS: Compared to Sham, only G8 (FXIII/DDAVP w/o hemorrhage) showed clotting time (CT) significantly lower (p<0.05). NBP alone (G6) resulted in significantly prolonged CT compared to G2, G3 and G5 (p<0.05). Similarly, median alpha angle was significantly larger in G3,4,5, and 9 compared to G6 (p<0.05). Area under the curve was significantly greater in G5 than G2. Intra-abdominal blood loss was lower in G5 and G9 compared to G2 and G6. FXIII/DDAVP and PH resulted in more robust fibrin mesh by EM. CONCLUSIONS: Normotensive resuscitation provokes more bleeding and worsens coagulation compared to pH, that is partially reversed by factor XIII and desmopressin. FXIII and DDAVP can synergistically improve coagulation. Permissive hypotension reduces bleeding regardless of those agents.
Assuntos
Desamino Arginina Vasopressina/uso terapêutico , Fator XIII/uso terapêutico , Hemostasia/efeitos dos fármacos , Hemostáticos/uso terapêutico , Ressuscitação/métodos , Choque Hemorrágico/tratamento farmacológico , Animais , Adesivo Tecidual de Fibrina/uso terapêutico , Hemodinâmica/efeitos dos fármacos , Masculino , Microscopia Eletrônica de Varredura , Coelhos , Distribuição Aleatória , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
It is reported the case of the use of cryoprecipitate in a canine patient, Yorkshire Terrier, who presented acute heavy bleeding disorder during arthroplasty procedure. At the time of bleeding disorder the patient had values of activated partial thromboplastin time (aPTT) above 240 seconds, and prothrombin time (PT) of 14.9 seconds. It was opted for the use of plasma components to correct the bleeding disorders, and since the patient had low weight (4.8kg), it was decided by the transfusion of one unit of cryoprecipitate (20 mL), quickly (20 minutes), even during surgery. The aPTT and PT post-transfusion values were 45.5 and 10.2 seconds respectively, with rapid control of bleeding. On return after 13 days the values of APTT and PT were within the normal range for the species. Because it contains high concentration of clotting factors and reduced volume, cryoprecipitate may be a good choice for patients with secondary bleeding disorders, especially in small animals. The effectiveness in transfusion therapy directed to the rational use of blood components illustrates the need for knowledge of the different blood components and individual study of each case.(AU)
Relata-se o caso do uso de crioprecipitado em um paciente canino, Yorkshire Terrier, que apresentou sangramento agudo intenso durante procedimento de artroplastia. No momento do distúrbio hemorrágico o paciente possuía valores de tromboplastina parcial ativada (TTPA) acima de 240 segundos, e de tempo de protrombina (TP) de 14,9 segundos. Optou-se pelo uso de hemocomponentes plasmáticos a fim de corrigir os distúrbios hemorrágicos, e uma vez que o paciente possuía peso reduzido (4,8kg), foi decidido pela transfusão de uma unidade de crioprecipitado (20 mL), de forma rápida (20 minutos), ainda no trans-operatório. Os valores de TTPA e TP pós-transfusionais foram de 45,5 e 10,2 segundos, respectivamente, com rápido controle do sangramento. No retorno após 13 dias os valores de TTPA e TP estavam dentro dos valores de normalidade para a espécie. Conclui-se que o crioprecipitado ofereceu os fatores de coagulação necessários para correção do distúrbio hemorrágico, controlando o sangramento e melhorando os valores de TTPA e TP. Por conter alta concentração de fatores de coagulação e volume reduzido, o crioprecipitado pode ser uma boa escolha para pacientes com coagulopatias secundárias, especialmente em animais de pequeno porte. A eficácia na terapia transfusional direcionada com o uso racional do melhor hemocomponente para o caso ilustra a necessidade de conhecimento dos diferentes hemocomponentes e estudo individual dos casos.(AU)
Assuntos
Animais , Cães , Hemorragia/veterinária , Hemostasia , Fator VIII/administração & dosagem , Fator XIII/administração & dosagem , Fibrinogênio/administração & dosagem , Artroplastia/veterinária , Transfusão de Componentes Sanguíneos/veterináriaRESUMO
This article presents the case of a young woman with massive hemoptysis (1,000 mL in 6 hours) due to tuberculosis, which could not be controlled by insertion of a Fogarty catheter through a fiber-optic bronchoscope. Because of asphyxia and persistent bleeding risk we instilled fibrinogen-thrombin through a fiber-optic bronchoscope inserted catheter, achieving bleeding cessation and permitting the placing of a double-lumen oro-tracheal tube. Later on, the patient underwent lobectomy and anti-tuberculosis treatment. The fibrinogen-thrombin could be considered as a bridge, transitory measure for massive hemoptysis, while definitive treatment could be established.