RESUMEN
We report a case of idiopathic hypereosinophilic syndrome (HES) presenting with cutaneous infarction and subsequent extensive deep vein thrombosis. The eosinophilia improved dramatically with systemic corticosteroid therapy. A variety of skin disorders have been associated with HES, although there are no previous reports of HES associated with cutaneous infarction. HES is a rare disorder characterized by a sustained overproduction of eosinophils and multisystem disease. The aetiology of the eosinophilia remains uncertain but clonal populations of abnormal T-cells producing interleukin 5 may be implicated.
Asunto(s)
Síndrome Hipereosinofílico/diagnóstico , Infarto/diagnóstico , Piel/irrigación sanguínea , Trombosis de la Vena/diagnóstico , Adulto , Estudios de Seguimiento , Glucocorticoides/uso terapéutico , Humanos , Síndrome Hipereosinofílico/tratamiento farmacológico , Masculino , Prednisolona/uso terapéuticoRESUMEN
BACKGROUND: A thrombotic etiology for inflammatory bowel disease (IBD) has been proposed as a result of its association with thromboembolic complications, smoking, the oral contraceptive pill, and the response of ulcerative colitis (UC) patients to heparin. We have previously demonstrated an increased prevalence of the Factor V Leiden mutation in UC and wished to investigate the frequency of the recently discovered prothrombin G20210A gene mutation in IBD. The aim of the study was to investigate the hypothesis that the prothrombic state associated with the prothrombin G20210A gene mutation is involved in the etiology of IBD. PATIENTS AND METHODS: A prospective cohort study of patients attending the Bristol Royal Infirmary and Gloucestershire Royal Hospital's IBD clinics was performed. Thirty-nine patients with IBD (24 with Crohn's disease and 15 with UC) and 100 historical controls were screened for the presence of the prothrombin gene mutation using a heteroduplex-based polymerase chain reaction technique. None of the patients with IBD had a personal history of thromboembolism, while three of them had a family history. RESULTS: No IBD patients had the prothrombin gene mutation compared with four (4%) controls (allelic frequency 2%). CONCLUSION: There does not appear to be an association of the prothrombin gene mutation with IBD and therefore it is unlikely to be involved in the etiology of IBD.
Asunto(s)
Colitis Ulcerosa/genética , Enfermedad de Crohn/genética , Mutación , Protrombina/genética , Estudios de Cohortes , Frecuencia de los Genes , Análisis Heterodúplex/métodos , Humanos , Estudios Prospectivos , Trombosis/genéticaRESUMEN
Peripheral T cell lymphocytosis is a rare finding in association with malignant thymomas. In the majority of previous cases, the tumours have behaved aggressively with symptoms arising from local invasion. We describe a patient with ocular myasthenia gravis who presented with a rapidly progressive polymyositis and neuropathy and who was subsequently found to have a thymic mass and a mild T cell lymphocytosis. The thymoma did not give rise to local symptoms and showed no evidence of progression over a 14-month period of follow-up. The possibility of an underlying thymic tumour should be considered in any patient with chronic T cell lymphocytosis if the circulating cells show mature morphology and there is no molecular evidence of monoclonality.
Asunto(s)
Linfocitosis/patología , Miastenia Gravis/patología , Polimiositis/patología , Linfocitos T/patología , Timoma/patología , Neoplasias del Timo/patología , Anciano , Humanos , MasculinoRESUMEN
CD56+ lymphomas derived from natural killer (NK) cell lineage are rarely encountered in Western populations and their clinical and pathological features have not been fully defined. The majority of reported cases are lymphomas of the nasal cavity, which are most commonly seen in Asia. A subtype of CD56+ lymphoma has recently been described (blastoid NK-cell lymphoma) which characteristically presents in older patients with cutaneous infiltrates and disease at other nodal and extranodal sites. We describe a case that correlates well with the clinicopathological features of blastoid NK-cell lymphoma. An unusual feature in our patient was that the cutaneous features of the lymphoma showed complete resolution shortly following commencement of oral steroid therapy.
Asunto(s)
Antiinflamatorios/uso terapéutico , Antineoplásicos Hormonales/uso terapéutico , Células Asesinas Naturales/patología , Linfoma Cutáneo de Células T/tratamiento farmacológico , Prednisolona/uso terapéutico , Anciano , Antígeno CD56/inmunología , Resultado Fatal , Humanos , Linfoma Cutáneo de Células T/inmunología , Linfoma Cutáneo de Células T/patología , Masculino , Inducción de RemisiónRESUMEN
BACKGROUND: A thrombotic aetiology for inflammatory bowel disease (IBD) has been proposed as a result of its association with thrombo-embolic complications, smoking, the oral contraceptive pill and the response of ulcerative colitis (UC) patients to heparin. The factor V Leiden (FVL) mutation is the commonest inherited risk factor for thrombo-embolism. AIM: The aim of the study was to investigate the hypothesis that the pro-thrombotic state associated with the FVL mutation is involved in the aetiology of IBD. PATIENTS AND METHODS: A prospective cohort study of patients attending the Bristol Royal Infirmary IBD outpatient clinic was performed. Fifty-four patients with IBD (30 with Crohn's disease (CD) and 24 with UC) and 55 historical controls were screened for the presence of FVL using the activated protein C (APC) ratio. Abnormal APC ratios were confirmed to be due to FVL using a heteroduplex-based polymerase chain reaction (PCR) technique. RESULTS: Five patients had the FVL mutation, compared to two controls. One of the patients was homozygous. Two of the patients had CD and three UC. The differences between controls and IBD patients was significant when the allelic frequency of the FVL mutation in patients with UC was compared with controls, with a risk ratio of 2.27, but with limited data. CONCLUSION: There appears to be a weak association between FVL and UC. This association is not strong enough to imply a causal relationship, but may be responsible for some of the thrombo-embolic complications.
Asunto(s)
Factor V/genética , Ligamiento Genético , Enfermedades Inflamatorias del Intestino/genética , Resistencia a la Proteína C Activada/genética , Adulto , Anciano , Alelos , Estudios de Cohortes , Colitis Ulcerosa/genética , Enfermedad de Crohn/genética , Femenino , Frecuencia de los Genes , Pruebas Genéticas , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Mutación , Reacción en Cadena de la Polimerasa , Estudios ProspectivosRESUMEN
Molecular studies have been performed on a Greek family with factor XIII-A subunit deficiency. The 15 exons of the A subunit gene were amplified by polymerase chain reaction and analysed by direct nucleotide sequencing. A homozygous single base insertion (1286insC) in exon 9 of the gene was identified in three affected family members. The insertion results in a frameshift and a premature stop signal a short distance downstream at codon 403. Any A subunit protein expressed is likely to be unstable and lack part of the catalytic core domain together with both beta barrel domains towards the C-terminal of the molecule. This study contributes to our knowledge of the mutational spectrum in patients with factor XIII-A deficiency.
Asunto(s)
Mutación del Sistema de Lectura , Transglutaminasas/genética , Elementos Transponibles de ADN/genética , Exones/genética , Femenino , Humanos , Masculino , LinajeRESUMEN
The majority of patients with type 2N von Willebrand disease (VWD type 2N) have mutations in the region of the von Willebrand factor (VWF) gene encoding the factor VIII binding domain of VWF. Two mutations predominate among VWD type 2N patients: G2811A and C2696T, which respectively bring about the amino acid substitutions R854Q and R816W in VWF. Several other mutations have been found in VWD type 2N, including T2701A (H817Q) and G2823T (C858F). We have developed a genetic test which permits rapid screening for these four mutations in a single polymerase chain reaction (PCR). The test employs induced heteroduplex formation using two universal heteroduplex generators, one of which detects G2811A (R854Q) and G2823T (C858F), the other detects C2696T (R816W) and T2701A (H817Q). The allele frequency of the common G2811A (R854Q) mutation was investigated in the local (S. Wales) population by examination of 216 VWF genes (108 individuals) and was found to be 0.01. The heteroduplex-based test additionally detected a novel candidate type 2N mutation, C2810T (R854W) and a previously described polymorphism, G2805A (R852Q). The polymorphism showed allele frequencies of 0.92 (G nucleotide) and 0.08 (A nucleotide) in the population study.
Asunto(s)
Factor VIII/metabolismo , Estructura Terciaria de Proteína , Enfermedades de von Willebrand/genética , Factor de von Willebrand/genética , Secuencia de Bases , Clonación Molecular , Estudios Transversales , Exones , Humanos , Datos de Secuencia Molecular , Mutación , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Enfermedades de von Willebrand/diagnósticoRESUMEN
We have shown previously that the Rhesus (Rh) polypeptides are the commonest targets for pathogenic anti-red blood cell (RBC) autoantibodies in patients with autoimmune hemolytic anemia (AIHA). The aim of the current work was to determine whether activated T cells from such patients also mount recall responses to epitopes on these proteins. Two panels of overlapping 15-mer peptides, corresponding to the sequences of the 30-kD Rh proteins associated with expression of the D and Cc/Ee blood group antigens, were synthesized and screened for the ability to stimulate the in vitro proliferation of mononuclear cells from the peripheral blood or spleen of nine AIHA cases. Culture conditions were chosen that favor recall proliferation by previously activated T cells, rather than primary responses. In seven of the patients, including all four cases with autoantibody to the Rh proteins, two or more peptides elicited proliferation, but cells from eight of nine patients with other anemias and seven of nine healthy donors failed to respond to the panels. Multiple peptides were also stimulatory in two positive control donors who had been alloimmunized with Rh D-positive RBCs. Six different profiles of peptides elicited responses in the AIHA patients, and this variation may reflect the different HLA types in the group. Stimulatory peptides were identified throughout domains shared between, or specific to, each of the related 30-kD Rh proteins, but T cells that responded to nonconserved regions did not cross-react with the alternative sequences. Anti-major histocompatibility complex class II antibodies blocked the responses and depletion experiments confirmed that the proliferating mononuclear cells were T cells. Notably, splenic T cells that proliferated against multiple Rh peptides also responded to intact RBCs. We propose that pathogenic autoantibody production in many cases of AIHA is driven by the activation of T-helper cells specific for previously cryptic epitopes on the Rh proteins.
Asunto(s)
Anemia Hemolítica Autoinmune/inmunología , Enfermedades Autoinmunes/inmunología , Epítopos/inmunología , Sistema del Grupo Sanguíneo Rh-Hr/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Adulto , Anciano , Secuencia de Aminoácidos , Anemia/inmunología , Autoanticuerpos/inmunología , Células Cultivadas , Femenino , Humanos , Inmunidad Celular , Memoria Inmunológica , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Isoinmunización Rh/inmunologíaRESUMEN
Hereditary haemochromatosis is an autosomal recessive disorder characterised by life-long excessive accumulation of iron. A candidate gene for hereditary haemochromatosis has recently been reported (HLA-H) and a specific missense mutation (Cys282Tyr) has been identified in 85% of patients with the disorder. We describe the rapid detection of this mutation using the polymerase chain reaction and restriction endonuclease digestion. The usefulness of this test for early diagnosis of hereditary haemochromatosis in asymptomatic family members is highlighted.
Asunto(s)
Genes MHC Clase I , Hemocromatosis/diagnóstico , Mutación , Cisteína/genética , Análisis Mutacional de ADN , Femenino , Hemocromatosis/genética , Humanos , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Tirosina/genéticaRESUMEN
Molecular analysis has been performed on a Malaysian patient with a severe bleeding disorder due to factor XIII(A) subunit deficiency. Total mRNA was isolated from the patient's leucocytes and four overlapping segments corresponding to the entire coding region of the A subunit cDNA were amplified by RT-PCR. The cDNA segments amplified efficiently and were of expected size. Direct sequencing of the complete reading frame revealed a single homozygous base change (nt 1327G-T) in exon 10 corresponding to a missense mutation, Val414Phe, in the catalytic core domain of the A subunit monomer. The mutation eliminates a BsaJ1 restriction site and family screening showed that both parents were heterozygous for the defect. The base substitution was absent in 55 normal individuals. Val414 is a highly conserved residue in the calcium-dependent transglutaminase enzyme family. Computer modelling based on 3D crystallographic data predicts that the bulky aromatic side chain of the substituted phenylalanine residue distorts protein folding and destabilizes the molecule. In addition, conformation changes in the adjacent catalytic and calcium binding regions of the A subunit are likely to impair the enzymatic activity of any protein synthesized.
Asunto(s)
Deficiencia del Factor XIII/genética , Factor XIII/genética , Mutación/genética , Adulto , Secuencia de Aminoácidos , Secuencia de Bases , ADN/genética , Exones/genética , Factor XIII/química , Humanos , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN/genética , Análisis de Secuencia de ADNAsunto(s)
Factor V/genética , Mutación , Técnicas Genéticas , Humanos , Sensibilidad y EspecificidadRESUMEN
Activated protein C resistance caused by factor V Leiden is an important thrombophilia disorder which predisposes to venous thromboembolism. Some studies also suggest a role in the pathogenesis of arterial thrombosis and atherosclerosis. The authors have investigated the prevalence of activated protein C resistance and factor V Leiden in a series of 45 patients with peripheral vascular disease. Twelve patients were receiving warfarin. The activated protein C resistance ratios were significantly lower in the group of 33 non-warfarinized patients with peripheral vascular disease (median 2.82 (range 1.36-3.83)) compared with 33 age- and sex-matched controls (median 2.97 range 2.24-4.11); P<0.005; Wilcoxon rank sum). Eight patients (24%) had activated protein C resistance (ratio <2.2). The prevalence of factor V Leiden in patients with peripheral vascular disease was 17.8% (8/45). This is significantly increased compared with the local population and UK published frequency of 3.5% for this genotype. The presence of factor V Leiden did not affect the late outcome of arterial reconstructive surgery in terms of graft patency (P=0.5, Fisher's Exact test).
Asunto(s)
Arteriosclerosis/sangre , Factor V/metabolismo , Oclusión de Injerto Vascular/sangre , Isquemia/sangre , Pierna/irrigación sanguínea , Proteína C/metabolismo , Anciano , Arteriosclerosis/cirugía , Pruebas de Coagulación Sanguínea , Activación Enzimática/efectos de los fármacos , Activación Enzimática/fisiología , Femenino , Oclusión de Injerto Vascular/cirugía , Humanos , Isquemia/cirugía , Masculino , Persona de Mediana Edad , Reoperación , Factores de Riesgo , Warfarina/administración & dosificaciónRESUMEN
A new genetic test has been developed for detection of the mutation known as factor V Leiden. The test employs heteroduplex technology and comprises a single PCR reaction followed immediately by PCR product analysis. It therefore represents the minimum practical route from blood/tissue sample to genetic result. A cohort of 100 patients with a history of thrombosis have been screened using both the new heteroduplex test and a previously described PCR-restriction endonuclease test. Results gave 100% correlation: normals 75 (75%), heterozygotes 24 (24%) and homozygotes 1 (1%). The heteroduplex test has been shown to give straightforward diagnosis in three different analytical systems: standard polyacrylamide gel electrophoresis (PAGE), mini-gel PAGE and capillary electrophoresis. The latter system is semiautomated, therefore rapid through-put of large sample numbers is now possible.
Asunto(s)
Factor V/genética , Técnicas Genéticas , Mutación , Humanos , Reacción en Cadena de la Polimerasa/métodosRESUMEN
We investigated the molecular basis of factor XIII(A) subunit deficiency in a Greek family. Each of the 15 exons of the A subunit gene were individually amplified by polymerase chain reaction, using previously reported oligoprimers. The proband with severe deficiency was found to have a homozygous 13-base pair deletion in the 3' half of exon 3. The deleted sequence, extending from codons 82-86, results in a frameshift and generates a downstream termination codon in exon 4. Single strand conformation polymorphism (SSCP) analysis detected no additional mutations in the coding or consensus splice sequences of the A subunit gene. Both parents of the proband were heterozygous for the defect. Only one previous microdeletion (AG dinucleotide) has been reported in the A subunit gene, and was located at the intron B-exon 3 boundary. Further studies are necessary to determine whether this region of the gene is a "hot spot" for microdeletion mutations.
Asunto(s)
Factor XIII/genética , Eliminación de Gen , Transglutaminasas/deficiencia , Secuencia de Aminoácidos , Secuencia de Bases , Niño , Cristalografía por Rayos X , Exones , Femenino , Homocigoto , Humanos , Masculino , Datos de Secuencia Molecular , Linaje , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple , Conformación Proteica , Transglutaminasas/genéticaRESUMEN
Residual donor leucocytes are responsible for many adverse transfusion reactions. Prestorage leucodepletion may ameliorate these effects and enhance product quality. We studied a bottom and top (BAT) system incorporating an integral filter for whole blood leucodepletion. Our evaluation assessed leucodepletion efficiency as well as in vitro SAG-M red cell quality and storage characteristics. Sixty-six units of blood were collected; test units into the Optipac-pLuS system and controls into the standard triple pack configuration. Test units were held for 4-6 h at room temperature (rt) or 12-18 h at 4 degrees C. The mean leucocyte counts for the SAG-M red cells in the quality and storage trial were 0.6 x 10(6) (rt hold), 0.05 x 10(6) (4 degrees C hold) and 2500 x 10(6) (controls). We observed no significant differences between the groups for Na+, ATP, 2,3-DPG, glucose, lactate and pH during the 49 d storage. The control group, however, showed a greater increase in haemolysis and K+ with time. Autologous in vivo 24 h red cell recovery, after 42 d storage, was > 75%. Adjustment of processing parameters in subsequent studies gave leucodepleted SAG-M red cells with minimal cell loss (9.19%) plus acceptable haemoglobin content (46-76 g/U) and haematocrit (54-62%). This system achieved > 3.5 log leucodepletion with all but one unit containing < 1 x 10(6) leucocytes. The product quality is good and the system suitable for routine use in blood centres.
Asunto(s)
Eliminación de Componentes Sanguíneos/métodos , Conservación de la Sangre/métodos , Separación Celular/métodos , Eritrocitos , Filtración , Humanos , LeucocitosRESUMEN
Activated protein C resistance resulting from Factor V Leiden is an important inherited thrombophilia disorder which is found in 3.5% of people in the UK. The genetic defect can be detected using the PCR and the diagnosis can be made postmortem from paraffin wax embedded tissue. The presence of Factor V Leiden should be sought in all cases of unexplained sudden death resulting from venous thromboembolism.