RESUMO
OBJECTIVES: This study aimed to assess the morphological differences in the articular disc (AD) between hemophilic patients and healthy individuals of the control group for further association with signs and symptoms. METHODS: Fourteen severe hemophilic patients had their AD evaluated by magnetic resonance imaging (MRI). The morphological findings were compared to those of a control group consisting of 14 healthy individuals. MRI was used to evaluate all the components of the temporomandibular (TMJ), including the AD, resulting in sequential T1-weighted parasagittal images. All the images were acquired with teeth in maximum intercuspation position. RESULTS: Morphological alterations showed significant statistical differences (P-value = 0.0068), whereas no statistical differences were found in the other variables, including TMJ pain, headache, bruxism and mouth opening limitation. In the group of non-hemophilic individuals, only two (14.29%) presented AD with non-biconcave features, whereas in the group of hemophilic patients, nine (64.29%) presented AD with a morphology other than biconcave. CONCLUSIONS: In patients with severe hemophilia, there seems to be a pattern of morphological alterations in the articular disc over time. The standard biconcave morphology of AD tends to change into other ones, particularly biplanar, hemiconvex and folded.
Assuntos
Hemofilia A , Transtornos da Articulação Temporomandibular , Humanos , Disco da Articulação Temporomandibular/diagnóstico por imagem , Hemofilia A/diagnóstico por imagem , Hemofilia A/patologia , Transtornos da Articulação Temporomandibular/diagnóstico por imagem , Transtornos da Articulação Temporomandibular/patologia , Articulação Temporomandibular , Imageamento por Ressonância Magnética/métodosRESUMO
BACKGROUND AND OBJECTIVES: Hemophilia A (HA) is an X-linked blood disorder. It is caused by pathogenic F8 gene variants, among which missense mutations are the most prevalent. The resulting amino acid substitutions may have different impacts on physicochemical properties and, consequently, on protein functionality. Regular prediction tools do not include structural elements and their physiological significance, which hampers our ability to functionally link variants to disease phenotype, opening an ample field for investigation. The present study aims to elucidate how physicochemical changes generated by substitutions in different protein domains relate to HA, and which of these features are more consequential to protein function and its impact on HA phenotype. METHODS: An in silico evaluation of 71 F8 variants found in patients with different HA phenotypes (mild, moderate, severe) was performed to understand protein modifications and functional impact. Homology modeling was used for the structural analysis of physicochemical changes including electrostatic potential, hydrophobicity, solvent-accessible/excluded surface areas, disulfide disruptions, and substitutions indexes. These variants and properties were analyzed by hierarchical clustering analysis (HCA) and principal component analysis (PCA), independently and in combination, to investigate their relative contribution. RESULTS: About 69% of variants show electrostatic changes, and almost all show hydrophobicity and surface area modifications. HCA combining all physicochemical properties analyzed was better in reflecting the impact of different variants in disease severity, more so than the single feature analysis. On the other hand, PCA led to the identification of prominent properties involved in the clustering results for variants of different domains. CONCLUSIONS: The methodology developed here enables the assessment of structural features not available in other prediction tools (e.g., surface distribution of electrostatic potential), evaluating what kind of physicochemical changes are involved in FVIII functional disruption. HCA results allow distinguishing substitutions according to their properties, and yielded clusters which were more homogeneous in phenotype. All evaluated properties are involved in determining disease severity. The nature, as well as the position of the variants in the protein, were shown to be relevant for physicochemical changes, demonstrating that all these aspects must be collectively considered to fine-tune an approach to predict HA severity.
Assuntos
Fator VIII/química , Hemofilia A , Fator VIII/genética , Fator VIII/metabolismo , Hemofilia A/genética , Hemofilia A/patologia , Humanos , Mutação , Mutação de Sentido Incorreto , Fenótipo , Eletricidade EstáticaRESUMO
Mechanisms thought to regulate activated factor VIII (FVIIIa) cofactor function include A2-domain dissociation and activated protein C (APC) cleavage. Unlike A2-domain dissociation, there is no known phenotype associated with altered APC cleavage of FVIII, and biochemical studies have suggested APC plays a marginal role in FVIIIa regulation. However, the in vivo contribution of FVIIIa inactivation by APC is unexplored. Here we compared wild-type B-domainless FVIII (FVIII-WT) recombinant protein with an APC-resistant FVIII variant (FVIII-R336Q/R562Q; FVIII-QQ). FVIII-QQ demonstrated expected APC resistance without other changes in procoagulant function or A2-domain dissociation. In plasma-based studies, FVIII-WT/FVIIIa-WT demonstrated dose-dependent sensitivity to APC with or without protein S, whereas FVIII-QQ/FVIIIa-QQ did not. Importantly, FVIII-QQ demonstrated approximately fivefold increased procoagulant function relative to FVIII-WT in the tail clip and ferric chloride injury models in hemophilia A (HA) mice. To minimize the contribution of FV inactivation by APC in vivo, a tail clip assay was performed in homozygous HA/FV Leiden (FVL) mice infused with FVIII-QQ or FVIII-WT in the presence or absence of monoclonal antibody 1609, an antibody that blocks murine PC/APC hemostatic function. FVIII-QQ again demonstrated enhanced hemostatic function in HA/FVL mice; however, FVIII-QQ and FVIII-WT performed analogously in the presence of the PC/APC inhibitory antibody, indicating the increased hemostatic effect of FVIII-QQ was APC specific. Our data demonstrate APC contributes to the in vivo regulation of FVIIIa, which has the potential to be exploited to develop novel HA therapeutics.
Assuntos
Fator VIII/metabolismo , Hemofilia A/patologia , Hemostasia , Proteína C/metabolismo , Proteínas Recombinantes/metabolismo , Animais , Cloretos/toxicidade , Fator VIII/genética , Feminino , Compostos Férricos/toxicidade , Hemofilia A/induzido quimicamente , Hemofilia A/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína C/genética , Proteínas Recombinantes/genéticaRESUMO
INTRODUCTION: Chronic pain is common in individuals with severe and moderate haemophilia who did not receive prophylaxis during childhood. OBJECTIVE: To verify the effectiveness of acupuncture in reducing intensity in chronic pain, changes in quality of life, joint function and impact on treatment satisfaction of haemophilia patients. METHODS: Single-blinded randomized clinical trial with 28 participants divided into two groups: Acupuncture (G1) treated with traditional unilateral acupuncture (side of greatest referred pain) and Control (G2) treated with transcutaneous electrical nerve stimulation (TENS), with electrodes on the joint of most intense pain. Both groups had a 20-minute session per week, total of 05 consecutive sessions. Before starting treatment, participants underwent sociodemographic assessment, physical assessment (HJHS), quality of life questionnaire (Haem-a-Qol) and treatment expectation (Likert scale). After the end of the fifth session, Haem-a-Qol, HJHS and degree of satisfaction (Likert) were performed. The assessment of pain intensity using the visual analogue scale (VAS) was performed before the beginning and after the end of all sessions in both groups. Statistical analysis was performed using ANOVA, Bonferroni, t test and chi-square test (P < .05). RESULTS: There was a statistical difference within and between groups G1 and G2 in reduction of VAS. In Haem-a-Qol, the groups showed similarity in quality of life. Both groups had high expectations for treatment. G1 presented a better degree of treatment satisfaction than G2. Total HJHS showed no difference within and between groups. CONCLUSION: Acupuncture was effective in reducing pain intensity in haemophilia patients with chronic joint disease when compared to TENS.
Assuntos
Terapia por Acupuntura/métodos , Dor Crônica/terapia , Hemofilia A/terapia , Adolescente , Adulto , Idoso , Dor Crônica/patologia , Feminino , Hemofilia A/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Hemophilia is a hereditary disorder that can be life-threatening in individuals who have severe spontaneous bleeding resulting from minor trauma or surgery. Although replacement therapy of the missing exogenous factor has improved patients' quality of life, it has not been possible to establish a long-term treatment. Due to the severity of the disease and the need for repetitive doses throughout the patient's life, replacement therapy has become a high-cost treatment option; therefore, the development of self-sustainable long-term therapies is critical. Hemophilia is a good candidate for gene therapy because it is a monogenic disease that can be counteracted by expression of the missing factor. In this article, we review some of the most relevant advances in gene therapy for this illness.
Assuntos
Terapia Genética , Hemofilia A/terapia , Hemorragia/genética , Vetores Genéticos/genética , Vetores Genéticos/uso terapêutico , Hemofilia A/genética , Hemofilia A/patologia , Hemorragia/patologia , Humanos , Qualidade de VidaRESUMO
INTRODUCTION: Haemophilic pseudotumour (HP) is an encapsulated haematoma in patients with haemophilia (PWH) which has a tendency to progress and produce clinical symptoms related to its anatomical location. AIM: To show the experience of one surgeon who has been using mini-invasive technique to treat pseudotumours of limbs in PWH with and without inhibitors at one centre for 28 years. MATERIALS AND METHODS: Thirty-three patients with 39 HP were treated. All patients had haemophilia A. Twenty-four patients had no inhibitors (72.8%), and 9 had inhibitors (27.2%). The mean follow-up was 16 years (1-25). All patients had x-rays and MRIs. All of them received Buenos Aires protocol as conservative treatment for 6 weeks. MRIs were repeated after 6 weeks' treatment to assess response to treatment. Surgery was performed in patients who did not respond to conservative treatment. RESULTS: After Buenos Aires protocol, four pseudotumours did not shrink (10.24%), 33 (84.61%) shrank, and two (5.12%) healed. Thirty-seven pseudotumours had surgery, 35 pseudotumours (94.59%) healed with minimally invasive treatment, and two did not heal (5.41%). No infection was observed with this treatment. The mortality rate for the series was 0%. CONCLUSION: The minimally invasive treatment of pseudotumours was effective in 95% of the cases and resulted in no mortality in this series after 28 years.
Assuntos
Extremidades/patologia , Hematoma/cirurgia , Hemofilia A/patologia , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Adolescente , Adulto , Criança , Coagulantes/administração & dosagem , Coagulantes/uso terapêutico , Tratamento Conservador/métodos , Extremidades/diagnóstico por imagem , Fator VIIa/administração & dosagem , Fator VIIa/uso terapêutico , Hematoma/tratamento farmacológico , Hemofilia A/complicações , Humanos , Infusões Intravenosas , Imageamento por Ressonância Magnética/métodos , Pessoa de Meia-Idade , Radiografia/métodos , Neoplasias de Tecidos Moles/diagnóstico por imagem , Neoplasias de Tecidos Moles/patologia , Resultado do Tratamento , Adulto JovemRESUMO
INTRODUCTION: People with haemophilic arthropathy (PWHA) have impairments in postural control. However, little is known about the effects of demanding conditions, including the unipedal stance and dual tasks, on postural control in PWHA. AIM: Determine the effects of performing dual tasks while in the one-leg stance on postural sway and postural control complexity in PWHA vs. healthy active (HAG) and non-active (HNAG) groups of individuals. METHODS: Fifteen PWHA and 34 healthy subjects (18 active and 16 non-active) were recruited. Vertical (V), mediolateral (ML) and anteroposterior (AP) centre of mass signals were acquired using a 3-axis accelerometer placed at the L3/L4 vertebrae of subjects as they performed the one-leg stance under single and dual-task conditions. Sway balance and the complexity of postural control were studied via root mean square (RMS) acceleration and sample entropy, respectively. Increased complexity of postural sway was attributed to increased automatism of postural control. RESULTS: RMS values for PWHA were higher than HAG under both conditions for the V and ML axes, and higher than HNAG under the dual-task condition for the ML axis. Sample entropy was lower in PWHA than healthy individuals under the dual-task condition for V and ML axes, and the single-task condition for the ML axis (P < .05). CONCLUSION: PWHA had poorer postural sway and decreased postural control complexity when performing a one-leg stance than healthy people, especially when the dual-task condition was applied. These results may help to design new approaches to assess and improve postural control in PWHA.
Assuntos
Hemofilia A/complicações , Artropatias/reabilitação , Equilíbrio Postural/fisiologia , Adulto , Estudos de Casos e Controles , Feminino , Hemofilia A/patologia , Humanos , Artropatias/etiologia , Masculino , Adulto JovemAssuntos
Fator VIII/genética , Hemofilia A/diagnóstico , Tolerância Imunológica , Inibidores dos Fatores de Coagulação Sanguínea/sangue , Inversão Cromossômica , Fator VIII/uso terapêutico , Feminino , Rearranjo Gênico , Genótipo , Hemofilia A/tratamento farmacológico , Hemofilia A/patologia , Humanos , Lactente , Íntrons , Cariótipo , LinhagemAssuntos
Fator VIII/genética , Hemofilia A/genética , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Alelos , Sítios de Ligação , Brasil , Fator VIII/antagonistas & inibidores , Fator VIII/metabolismo , Frequência do Gene , Mutação em Linhagem Germinativa , Hemofilia A/patologia , Humanos , Masculino , MicroRNAs/genética , Fenótipo , Polimorfismo de Nucleotídeo Único , Índice de Gravidade de DoençaRESUMO
A hemofilia é uma doença que afeta a coagulação do sangue pela falta ou diminuição do fator de coagulação VIII ou IX. Esta deficiência faz com que a pessoa sangre por um tempo maior do que uma pessoa normal se não for medicada. Foi avaliado um indivíduo do sexo masculino de 31 anos com hemofilia A grave, artropatia hemofílica em cotovelo esquerdo e tornozelo direito. Foram realizadas 20 meses de treinamento resistido e aeróbio. As musculaturas envolvidas nos exercícios resistidos foram o peitoral maior, latíssimo do dorso, bíceps braquial, tríceps braquial, deltoide, quadríceps e isquiotibiais, sendo realizadas duas séries de 10 repetições para cada um e intervalo de 45 segundos, com intensidade de acordo com o teste inicial de 10 RM baseados na percepção de esforço de 11 a 13 da Escala de Borg. O exercício aeróbio foi realizado em bicicleta ergométrica horizontal com duração de 20 minutos, sendo aferida a frequência cardíaca de repouso, aos 10 e aos 20 minutos do exercício, e após três minutos do término. Nos seis meses antes de iniciar o programa o paciente sofreu três hemorragias, duas espontâneas, em cotovelo e tornozelo esquerdos e uma em coxa direita por pequeno trauma não identificado - em todas fazendo uso do FAH. Durante os 20 meses, o paciente teve uma hemorragia após oito meses de tratamento, no cotovelo esquerdo por trauma durante os exercícios após aumento de carga. Após este episódio, o paciente teve outro sangramento, porém espontâneo, na mesma articulação 12 meses depois. O menor ganho de força foi de tríceps braquial com 33% e o maior foi 257% em extensores de joelho, sendo a média de ganho geral de força muscular de 121%. A prática de exercícios físicos de forma supervisionada é um importante instrumento auxiliar no tratamento das pessoas com hemofilia, demonstrando a necessidade de treino de força e resistência muscular específico para este grupo de pessoas quanto à prevenção de lesões, evitando o desgaste e comprometimento do sistema musculoesquelético. O propósito deste estudo é apresentar os resultados obtidos em um programa de Condicionamento Físico, na prevenção de hemorragias de uma pessoa com hemofilia A grave e sua ação profilática, sem a administração do fator anti-hemofílico (FAH)
Hemophilia is a blood clotting disorder that causes the decrease or absence of blood coagulation factors VIII or IX. This disease causes a person to bleed longer than a normal person if it is not treated. A 31-year-old male with hemophilic arthropathy in the left elbow and right ankle was evaluated. The program lasted 20 months, with aerobic and muscle strength training. The muscles involved were the pectoralis major, latissimus dorsi, biceps brachii, triceps brachii, deltoideus, quadriceps femoris, and hamstring. The patient performed two series of 10 repetitions with 45 seconds between series. The intensity was based on the initial test of 10 maximum repetitions in accordance with the rate of perceived exertion on the Borg Scale. Aerobic exercise was performed on a horizontal stationary bike for 20 minutes. Pulse rate was registered at rest, after 10 and then 20 minutes of workout, and again after three minutes of recovery. Six months before the initiation of the fitness program the patient suffered three hemorrhages: two spontaneous in the left elbow and left ankle and one in the right leg due to a small unspecified trauma. All three hemorrhages were treated by means of clotting factors (CF's). During the program period, the first hemorrhage treated with CF's occurred in the left elbow by trauma after increasing the exercise load. Twelve months later, the patient had spontaneous bleeding in the same joint. The least improvement in strength was found for the triceps brachii with 33%, whereas the knee extensors improved the most with 257%. The average muscle strength increase was 121%. Monitored physical exercise is an important vehicle in treating of people with hemophilia, presenting the need for resistance training specific for hemophilic patients to prevent injuries and avoiding early wear of the musculoskeletal system. The objective of this study was to present the results of a physical activity program to prevent bleeding episodes in a person with severe hemophilia A and the prophylaxis without administering any clotting factors
Assuntos
Humanos , Exercício Físico , Força Muscular , Condicionamento Físico Humano , Hemofilia A/patologiaRESUMO
INTRODUCTION: The development of factor VIII (FVIII) inhibitor is the main complication of replacement therapy in patients with haemophilia A (HA). A ratio of 5-7% of individuals HA develops antibodies (inhibitors) against the FVIII infused during the treatment, thereby reducing their pro-coagulant activity. The immunomodulatory cytokine genes have been related to the risk of development of alloantibodies in several studies, mainly in HA with severe form. AIM: We investigated the polymorphisms in regulatory regions of cytokine genes (IL1A, IL1B, IL1R, IL1RA, IL4RA, IL12, INFG, TGFB1, TNF, IL2, IL4, IL6, IL10) that could influence the risk of developing inhibitors in patients with severe HA. METHODS: The genotyping of cytokine genes of 117 patients with HA was performed by polymerase chain reaction with sequence-specific primers (PCR-SSP) using the protocol recommended by the manufacturer (Invitrogen kit Cytokines(®) , Canoga Park, USA) RESULTS: From the cohort of 117 patients with severe HA, 35 developed inhibitors. There was a higher frequency of +874 T allele in INFG and of +869 TT and TG/TG in TGFB1 genes on patients with inhibitors. CONCLUSION: This suggests that polymorphisms in INFG and in TGFB1 genes are related to risk of developing inhibitor, and could contribute to a genetic profile of the individual HA for the risk of inhibitors development to FVIII.
Assuntos
Inibidores dos Fatores de Coagulação Sanguínea/sangue , Hemofilia A/genética , Interferon gama/genética , Fator de Crescimento Transformador beta1/genética , Adolescente , Adulto , Idoso , Alelos , Criança , Pré-Escolar , Fator VIII/imunologia , Fator VIII/uso terapêutico , Frequência do Gene , Genótipo , Haplótipos , Hemofilia A/tratamento farmacológico , Hemofilia A/patologia , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Índice de Gravidade de Doença , Adulto JovemRESUMO
In contrast to haemophilia B, allergic manifestations are rare complications in haemophilia A (HA) patients treated with factor VIII (FVIII) concentrates. Nevertheless, it can be serious and hamper replacement therapy in these cases. The aims of this study were to evaluate the frequency of allergic reaction in a cohort of HA patients treated only with plasma-derived FVIII (pdFVIII) concentrates, and assess the possible immune mechanisms involved. History of allergic reaction was retrospectively assessed. Patients with allergic manifestations were followed, and had plasma samples collected in different timepoints in relation to the allergic episode. These samples were analysed for the presence of inhibitor and anti-FVIII immunoglobulins subclasses. Three of 322 HA patients (0.9%) developed allergic reaction after exposure to pdFVIII products during the last 15 years in our centre. The first patient, with severe HA, without inhibitor, had anti-pdFVIII IgE and IgG4, but no anti-recombinant FVIII (rFVIII) IgE. The second patient, with severe HA, and high-responding inhibitor, presented allergic manifestation with both, pdFVIII concentrate and activated prothrombin complex concentrate. Although anti-pdFVIII and anti-rFVIII IgG4 were detected, no anti-FVIII IgE was present. The third patient, with moderate HA without inhibitor, atopic, had no anti-FVIII immunoglobulin detected, and allergic symptoms disappeared after switching to rFVIII concentrate. This study corroborates the low incidence of allergic reactions in HA patients. In the three cases presented, the anti-FVIII immunoglobulin profile demonstrated that the allergic manifestation was triggered by other proteins contained in pdFVIII products, and not directed to FVIII.
Assuntos
Fator VIII/uso terapêutico , Hemofilia A/tratamento farmacológico , Adulto , Pré-Escolar , Coagulantes/efeitos adversos , Coagulantes/imunologia , Coagulantes/uso terapêutico , Estudos de Coortes , Ensaio de Imunoadsorção Enzimática , Fator VIII/efeitos adversos , Fator VIII/imunologia , Hemofilia A/patologia , Humanos , Hipersensibilidade/etiologia , Hipersensibilidade/patologia , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Índice de Gravidade de DoençaRESUMO
Haemophilia and its treatment interfere with patients' life and may affect adherence to treatment. This study explored the impact of severe haemophilia A on patients' health status, especially in young adults (YA), using data from guardian(™) 1, a multinational, open-label, non-controlled phase 3 trial investigating safety and efficacy of turoctocog alfa (NovoEight(®) ) in previously treated patients aged 12 years and older with severe haemophilia A (FVIII ≤ 1%). Health status was assessed using the EuroQoL-5 dimensions (EQ-5D-3L), covering 5 dimensions of health (mobility, self-care, usual activities, pain/discomfort and anxiety/depression), and a visual analogue scale (VAS) measuring self-rated overall health status. EQ-5D was administered pretreatment (screening/baseline) and posttreatment (end-of-trial). Baseline responses to the EQ-5D dimensions and VAS were described overall and by age and compared to reference values from UK general population. Guardian(™) 1 included 150 patients (16 adolescents, 83 YA aged 16-29 and 51 adults aged 30+). All five dimensions of patients' health status were impacted at baseline. The percentage of haemophilia patients reporting problems was consistently significantly greater than age-matched general population reference values. Likewise, for all age groups mean baseline EQ-5D VAS score was significantly lower for haemophilia patients (YA: 78.0) than for the general population (YA aged 18-29: 87.3). The health status of patients with severe haemophilia A entering guardian(™) 1 was markedly poorer than that of the general population, particularly regarding mobility and pain. YA patients reported better health status than older patients, but considerably lower than that of the general YA population.
Assuntos
Fator VIII/uso terapêutico , Nível de Saúde , Hemofilia A/tratamento farmacológico , Adolescente , Adulto , Criança , Hemofilia A/economia , Hemofilia A/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Qualidade de Vida , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
The production of factor VIII inhibitor antibodies remains the most costly and serious complication in replacement therapy of hemophilia A. We investigated the clinical significance of CD4(+)CD25(high) T regulatory (Treg) cells in hemophilia patients. Our trial included 6 severe hemophilia A patients with factor VIII inhibitors, 6 hemophilia patients without inhibition of factor VIII, and 6 healthy persons (controls). Plasma factor VIII: c was measured by clotting assay. Peripheral blood samples were examined using mutiparameter flow cytometry with fluorescent-labeled monoclonal antibodies. Plasma levels of IFN-γ, IL-2, IL-10, and TGF-ß were measured by ELISA. The frequency of CD4(+)CD25(high) Treg cells in CD4(+) cells was 1.07 ± 0.38% in inhibitor patients and 0.57 ± 0.14% in non-inhibitor patients. The proportion of Treg cells in healthy controls was similar to that of the non-inhibitor patients. However, there were significant differences between the inhibitor and non-inhibitor patients in levels of IFN-γ, IL-2, IL-10, and TGF-ß. We conclude that the proportions of Treg cells and the concentrations of T cell cytokines in inhibitor patients are higher than those in non-inhibitor patients. The increased number of Treg cells and increased T-cell cytokines may be related to the development and efficiency of the factor VIII inhibitor.
Assuntos
Linfócitos T CD4-Positivos/metabolismo , Fator VIII/antagonistas & inibidores , Hemofilia A/tratamento farmacológico , Hemofilia A/genética , Linfócitos T Reguladores/metabolismo , Fator VIII/administração & dosagem , Fator VIII/imunologia , Citometria de Fluxo , Hemofilia A/patologia , Humanos , Imunofenotipagem , Interleucina-2/genética , Interleucina-2/metabolismo , Subunidade alfa de Receptor de Interleucina-2/genética , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/patologia , Ativação Linfocitária/genéticaRESUMO
A total of 76 unrelated male patients with mild (n = 55) or moderate (n = 21) haemophilia A living in the southern Brazilian state of Rio Grande do Sul were studied by direct sequencing of all F8 26 exons, the 5' UTR and 3' UTR, intron-exon junctions and the promoter region. When no mutation was found, a multiplex ligation-dependent probe amplification analysis was performed. We identified the disease-causing mutations in 69 patients, who showed 33 different mutations: 27 missense, one small deletion, two small duplications and three splice site mutations. Seven missense and two splice site mutations were not previously reported in HAMSTeRS and were not identified in any current literature search. Nine recurrent mutations were found, one of them never described before (p.Tyr1786Phe). Haplotype analysis indicated that this mutation had originated in the Brazilian population as a single event in a common ancestor. The possible influence of these mutations in the determination of the disease was carefully considered, including bioinformatic tools. These data add to the general knowledge of the disease and can also be useful for HA diagnosis and detection of carriers in the southern Brazilian population.
Assuntos
Fator VIII/genética , Hemofilia A/genética , Hemofilia A/patologia , Mutação , Regiões 3' não Traduzidas , Regiões 5' não Traduzidas , Adolescente , Adulto , Idoso , Brasil , Criança , Pré-Escolar , Éxons , Genótipo , Haplótipos , Hemofilia A/tratamento farmacológico , Humanos , Lactente , Íntrons , Masculino , Pessoa de Meia-Idade , Patologia Molecular , Fenótipo , Adulto JovemRESUMO
Haemophilia A is a hereditary bleeding disorder linked to the X chromosome characterized by a deficiency or defect in the coagulation factor VIII (FVIII). Individuals with this coagulopathy require constant infusions of FVIII to maintain their physical integrity and haemostasis. During treatment, some patients develop an immune response that produces antibodies to FVIII, also called inhibitors, affecting the pro-coagulant activity of this protein. Despite the clinical relevance of FVIII inhibitors, the immune mechanisms that lead to their production are not known. This study investigated the immunological cytokine profile using plasma from HA patients which were either positive or negative for FVIII inhibitors and from healthy individuals. The results showed that healthy individuals and HA patients that do not develop FVIII inhibitors have a mixed immune response profile with high secretion of IFN-γ, TNF-α IL-2 and IL-5. In contrast, HA patients with FVIII inhibitors exhibited an anti-inflammatory/regulatory immune response characterized by low levels of all measured cytokines except for IL-4 and IL-10. This profile may be related to the development and maintenance of the FVIII inhibitors. By comparing the cytokine profiles of the three different groups we have established a model explaining the immune activation resulting in the production of FVIII inhibitors in haemophilia A patients.
Assuntos
Inibidores dos Fatores de Coagulação Sanguínea/sangue , Citocinas/sangue , Fator VIII/antagonistas & inibidores , Hemofilia A/metabolismo , Adolescente , Adulto , Criança , Pré-Escolar , Fator VIII/metabolismo , Fator VIII/uso terapêutico , Hemofilia A/tratamento farmacológico , Hemofilia A/patologia , Humanos , Interferon gama/sangue , Interleucina-10/sangue , Interleucina-2/sangue , Interleucina-4/sangue , Interleucina-5/sangue , Masculino , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/sangue , Adulto JovemAssuntos
Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/metabolismo , Fator VIII/metabolismo , Glicoproteínas/metabolismo , 8-Bromo Monofosfato de Adenosina Cíclica/farmacologia , Animais , Asparagina/metabolismo , Bovinos , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , Chaperona BiP do Retículo Endoplasmático , Células Endoteliais/citologia , Fator VIII/genética , Glicoproteínas/genética , Glicosilação/efeitos dos fármacos , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Hemofilia A/genética , Hemofilia A/metabolismo , Hemofilia A/patologia , Humanos , Insulina/farmacologia , Neovascularização Patológica/metabolismo , Neovascularização Fisiológica/efeitos dos fármacos , Estrutura Terciária de Proteína , Transdução de Sinais/efeitos dos fármacos , Tunicamicina/farmacologiaRESUMO
JUSTIFICATIVA E OBJETIVOS: Há mais de 25 anos não se discute, na Revista Brasileira de Anestesiologia, de maneira geral, o manuseio do paciente hemofílico durante o peri-operatório. Apesar da hemofilia ter sido definida como doença no início do século XIX, existem, até hoje, muitas descobertas relacionadas a ela. O objetivo dessa revisão é apontar os cuidados relacionados ao paciente hemofílico durante o período peri-operatório, ressaltando o manuseio da hemofilia tipo A e o papel do anestesiologista na equipe multidisciplinar. CONTEUDO: Estão definidas as características da hemofilia quanto à clínica e aos achados laboratoriais, a terapia farmacológica atual e os cuidados com o manuseio do paciente hemofílico no intra-operatório. CONCLUSÕES: O manuseio do paciente hemofílico foi aprimorado. Como conseqüência desse avanço, percebe-se a importância e a necessidade de que novos conhecimentos, principalmente em relação à terapia de reposição, sejam dominados por hematologistas e por todo o corpo clínico-cirúrgico. Quando pacientes hemofílicos submetem-se a procedimentos cirúrgicos é necessário o envolvimento de uma equipe multidisciplinar da qual o anestesiologista faz parte. A este profissional cabe a responsabilidade de tomar as condutas mais adequadas frente ao paciente hemofílico, participando e comunicando-se ativamente com os membros da equipe multidisciplinar.
Assuntos
Humanos , Período Pós-Operatório , Período Pré-Operatório , Hemofilia A/patologia , Anestesia/métodosRESUMO
Besides intron 22 factor VIII gene inversion (Inv22), intron 1 inversion (Inv1) has recently been reported as a further recurrent mutation that causes approximately 5% of severe haemophilia A (HA) cases. We analysed the presence of the Inv1 in a group of 64 severe HA-affected families from Argentina, and found only one positive case. This Inv1 patient has not developed a factor VIII inhibitor, and the screening for small mutations in the coding sequences of the factor VIII gene did not detect any additional defect in this case. The Inv1 genotyping was further applied to analyse the haemophilia carrier status of the proband's sister. In addition, we studied the accuracy of the current polymerase chain reaction-based method to investigate the Inv1, and confirmed the absence of amplimer length polymorphisms associated to the Inv1-specific polymerase chain reaction amplifications in 101 X-chromosome haplotypes from unrelated Argentinian healthy males. In order to discuss Inv1 mutation frequency in severe HA and the risk of inhibitor formation, a review of the literature was included. Our data highlight the importance of analysis of the Inv1 in Inv22-negative severe HA cases. This will benefit both genetic counselling and the study of the relationship between genotype and inhibitor development.