RESUMEN
Marfan syndrome (MFS) is an inherited disorder of connective tissue due to mutations in FBN1 (90%) and TGFBR1 and TGFBR2 (5 to 10%) genes. Clinical and differential diagnosis is difficult because of the inter- and intrafamiliar marked heterogeneity and the variable onset age of clinical manifestations. Among the disorders, in differential diagnosis, thoracic aortic aneurysm (TAA) and Ullrich scleroatonic muscular dystrophy (UCMD) are reported. We evaluate the possibility of utilizing autofluorescence (AF) analysis as a diagnostic tool in the clinical and/or differential diagnosis of MFS and related disorders and in the investigation of the molecular mechanisms involved. Both multispectral imaging autofluorescence microscopy (MIAM) and autofluorescence microspectroscopy (AMS) have been used to characterize AF emission of fibroblasts from patients affected by inherited connective tissue disorders. Our preliminary results show significant differences in AF emission between normal and pathological fibroblasts, suggesting possible improvement in diagnostics of connective tissue disorders by AF analysis.
Asunto(s)
Fibroblastos/patología , Proteínas Luminiscentes/análisis , Síndrome de Marfan/diagnóstico , Síndrome de Marfan/metabolismo , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Espectrometría de Fluorescencia/métodos , Adulto , Biomarcadores/análisis , Células Cultivadas , Preescolar , Diagnóstico Diferencial , Femenino , Predicción , Humanos , Lactante , Masculino , Proyectos PilotoRESUMEN
PURPOSE: Ectopia lentis (EL) and Marfan syndrome (MFS) are considered two distinct clinical entities. We performed genetic and clinical studies to investigate whether EL is actually distinct from MFS or if it is a mild phenotypic expression of it. METHODS: Seven patients with EL were followed for 5-10 years. Mutation screening analysis of the 65 exons of FBN1 was performed by polymerase chain reaction (PCR) amplification of genomic DNA, denaturing high pressure liquid chromatography analysis, and direct sequencing of heteroduplexes. RESULTS: Yearly examinations during the 10 years of follow-up allowed the detection of a late onset of dural ectasia in six out of seven patients (age range: 32-64 years versus 8-55 years in MFS previously reported). We also detected the onset of mild thoracic aortic dilatation in a sporadic case (age 45). Six out of seven index cases of EL turned out to be mild forms of Marfan syndrome with possible late cardiovascular involvement as detected in one case. Four novel missense mutations and one known splicing mutation were detected in five out of seven (71%) patients. Their localization confirmed the presence of a first hot spot within exons 1-15 and suggested the presence of a second one between exons 31-39. CONCLUSIONS: The presence of a second major criterion in six EL patients shifted the clinical diagnosis from EL to MFS. These data demonstrate that some cases, which are initially diagnosed as EL, turn out to be mild Marfan patients. A clinical cardiovascular follow-up is therefore highly recommended for all EL patients since they may develop thoracic aortic aneurysm (TAA) or dissection later in life. Also magnetic resonance imaging (MRI) for dural ectasia (DE) should be performed in a complete follow up for a MFS diagnosis.
Asunto(s)
Desplazamiento del Cristalino/complicaciones , Síndrome de Marfan/complicaciones , Adolescente , Adulto , Niño , Desplazamiento del Cristalino/genética , Femenino , Fibrilina-1 , Fibrilinas , Estudios de Seguimiento , Humanos , Masculino , Proteínas de Microfilamentos/genética , Persona de Mediana Edad , Linaje , Fenotipo , Factores de TiempoRESUMEN
OBJECTIVES: Myocardial fibrosis is a major component of ventricular remodeling after large myocardial infarction (MI). The present study tests the hypothesis that post-ischemic myocardial fibrosis can occur independent of hemodynamic changes. METHODS: A mouse model of distal left coronary artery ligation was established to induce a small infarct (less than 15% of the left ventricle) in order to avoid significant mechanical overload after permanent myocardial ischemia. Left heart catheterization was performed to evaluate the post-infarct hemodynamics. Tissues from both ischemic and non-ischemic myocardium were examined for mRNA and protein expression at 24, 72 h and 7 days after ligation. RESULTS: Heart/body weight ratio after ligation was increased by approximately 10% over sham control although there is no statistically significant difference in hemodynamic parameters between the two groups. Non-ischemic myocardium distant from the infarct site showed molecular evidence of myocardial fibrosis 72 h and 7 days after ligation. There was marked up-regulation of mRNAs for extracellular matrix (ECM) proteins and their cross-linking enzyme, such as collagens type I, III and VI, and lysyl oxidase. Immunohistochemical study confirmed that the expression of these ECM proteins was significantly increased in the non-ischemic myocardium after 7 days. TGF-beta1 was up-regulated after 72 h in both ischemic and non-ischemic myocardium. CONCLUSIONS: Molecular and histopathological findings demonstrate that abnormal myocardial fibrosis can be induced by a small infarct independent of secondary hemodynamic changes.
Asunto(s)
Hemodinámica , Isquemia Miocárdica/patología , Remodelación Ventricular , Animales , Colágeno/análisis , Proteínas de la Matriz Extracelular/análisis , Fibrosis , Frecuencia Cardíaca , Inmunohistoquímica/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Miocardio/patología , Tamaño de los Órganos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Volumen Sistólico , Factores de Tiempo , Factor de Crecimiento Transformador beta/análisisRESUMEN
BACKGROUND: The management of pT1a-b pN0 HER2-positive breast cancer is controversial and no data about the efficacy of trastuzumab in this setting are available from randomized clinical trials. The aims of this retrospective study were to assess how patients are managed in clinical practice in Italy, which clinical or biological characteristics influenced the choice of adjuvant systemic therapy and the outcome of patients. METHODS: Data of consecutive patients who underwent surgery from January 2007 to December 2012 for HER2-positive, pT1a-b pN0 M0 breast cancer were retrospectively collected from 28 Italian centres. Analysis of contingency tables and multivariate generalized logit models were used to investigate the association between the baseline clinical and biological features and the treatment strategy adopted. RESULTS: Among 303 enrolled patients, 204 received adjuvant systemic therapy with trastuzumab, 65 adjuvant systemic therapy without trastuzumab and 34 did not receive adjuvant systemic therapy. At the multivariate analysis age, tumor size, proliferation index and hormone receptor status were significantly associated with the treatment choice. Five-year disease-free survival (DFS) probability was 95%, 94.3% and 69.6% for patients treated with adjuvant systemic therapy and trastuzumab, with adjuvant systemic therapy without trastuzumab and for patients who did not receive adjuvant systemic therapy, respectively (p<0.001). CONCLUSIONS: The majority of patients (66%) with pT1a-b pN0 HER2-positive breast cancer enrolled in this retrospective study received adjuvant systemic therapy with trastuzumab, whereas only 11% patients did not receive any adjuvant systemic therapy. It should be emphasized, however, that in the adjuvant systemic therapy without trastuzumab group 94% of tumors were hormone receptor positive and 89% of patients were treated with endocrine therapy only [corrected]. The 5-year DFS probability was significantly higher for patients receiving adjuvant systemic therapy with trastuzumab compared with patients not receiving adjuvant systemic therapy or receiving adjuvant systemic therapy without trastuzumab.
Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Carcinoma Ductal de Mama/tratamiento farmacológico , Receptor ErbB-2/genética , Trastuzumab/uso terapéutico , Adyuvantes Farmacéuticos/uso terapéutico , Adulto , Factores de Edad , Anciano , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/cirugía , Carcinoma Ductal de Mama/diagnóstico , Carcinoma Ductal de Mama/mortalidad , Carcinoma Ductal de Mama/cirugía , Quimioterapia Adyuvante , Manejo de la Enfermedad , Femenino , Expresión Génica , Humanos , Persona de Mediana Edad , Análisis Multivariante , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Pronóstico , Receptor ErbB-2/metabolismo , Estudios Retrospectivos , Análisis de Supervivencia , Carga TumoralRESUMEN
By mRNA differential display in mouse hearts, lysyl oxidase (Lox), a key enzyme catalyzing cross-links in elastin and collagens, was found to be up-regulated between embryonic days 11 (E11) and 13 (E13). This was confirmed by semiquantitative RT-PCR. We analyzed its spatio-temporal expression pattern by in situ hybridization in regard to the development of myocardial cells, endocardial cushion tissue, aortic arch vessels, and epicardium.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Corazón/embriología , Proteína-Lisina 6-Oxidasa/metabolismo , Animales , Corazón/crecimiento & desarrollo , Ratones , Regulación hacia ArribaRESUMEN
INTRODUCTION: Moderate hyperhomocysteinemia is considered a risk factor for both venous and arterial thrombosis. A prevalence of up to 30% of fasting hyperhomocysteinemia has been recently reported in patients with retinal vein occlusion (RVO) whereas conflicting data exist on the role of C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene as a risk factor for RVO. No report has been published on cystathionine beta-synthase (CBS) 844ins68 polymorphism (another genetic determinant of blood Hcy levels) in RVO patients. Moreover, scarce information is available on the usefulness of measuring homocysteine also after methionine loading to increase the diagnostic efficacy of hyperhomocysteinemia in RVO patients. MATERIALS AND METHODS: In 55 consecutive patients with diagnosis of RVO and 65 matched controls, plasma fasting total homocysteine (Hcy) levels and CBS and MTHFR polymorphisms were evaluated. In patients with normal fasting Hcy levels, post-methionine Hcy levels were determined. RESULTS: Moderate fasting hyperhomocysteinemia was detected in 18/55 patients (32.7%). In the remaining 37 patients, Hcy was measured again post-methionine loading (PML). Only 3/37 (8.1%) patients had PML hyperhomocysteinemia. Thus, the total prevalence of moderate hyperhomocysteinemia in this cohort of RVO patients was 21/55 (38.2%). The prevalence of homozygosity for C677T MTHFR genotype, but not that of heterozygosity for CBS844ins68, was significantly higher in RVO patients than in controls. CONCLUSIONS: Differently from what has been reported for arterial and/or venous thrombosis, a single fasting Hcy measurement is able to detect most of RVO patients (85.7%) with moderate hyperhomocysteinemia. C677T MTHFR, but not CBS 844ins68, genotype may play a role as risk factor for RVO.
Asunto(s)
Cistationina betasintasa/genética , Hiperhomocisteinemia/genética , Metionina , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Oclusión de la Vena Retiniana/sangre , Trombofilia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Sustitución de Aminoácidos , Ayuno/sangre , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Hiperhomocisteinemia/complicaciones , Masculino , Metionina/farmacología , Persona de Mediana Edad , Mutagénesis Insercional , Mutación Missense , Mutación Puntual , Polimorfismo Genético , Oclusión de la Vena Retiniana/etiología , Oclusión de la Vena Retiniana/genética , Factores de Riesgo , Trombofilia/complicacionesRESUMEN
Marfan syndrome is an autosomal dominant disorder of connective tissue caused by mutations in the gene encoding fibrillin-1 (FBN1), a matrix component of microfibrils. Dural ectasia, i.e. enlargement of the neural canal mainly located in the lower lumbar and sacral region, frequently occurs in Marfan patients. The aim of our study was to investigate the role of dural ectasia in raising the diagnosis of Marfan syndrome and its association with FBN1 mutations. We studied 40 unrelated patients suspected for MFS, who underwent magnetic resonance imaging searching for dural ectasia. In all of them FBN1 gene analysis was also performed. Thirty-seven patients resulted affected by Marfan syndrome according to the '96 Ghent criteria; in 30 of them the diagnosis was confirmed when revaluated by the recently revised criteria (2010). Thirty-six patients resulted positive for dural ectasia. The degree of dural ectasia was grade 1 in 19 patients, grade 2 in 11 patients, and grade 3 in 6 patients. In 7 (24%) patients, the presence of dural ectasia allowed to reach a positive score for systemic feature criterion. Twenty-four patients carried an FBN1 mutation, that were represented by 13 missense (54%), and 11 (46%) mutations generating a premature termination codon (PTC, frameshifts and stop codons). No mutation was detected in the remaining 16 (6 patients with MFS and 10 with related disorders according to revised Ghent criteria). The prevalence of severe (grade 2 and grade 3) involvement of dura mater was higher in patients harbouring premature termination codon (PTC) mutations than those carrying missense-mutations (8/11 vs 2/13, P = 0.0111). Our data emphasizes the importance of dural ectasia screening to reach the diagnosis of Marfan syndrome especially when it is uncertain and indicates an association between PTC mutations and severe dural ectasia in Marfan patients.
Asunto(s)
Síndrome de Marfan/diagnóstico , Proteínas de Microfilamentos/genética , Mutación , Defectos del Tubo Neural/diagnóstico , Médula Espinal/patología , Adolescente , Adulto , Niño , Dilatación Patológica , Femenino , Fibrilina-1 , Fibrilinas , Pruebas Genéticas , Humanos , Imagen por Resonancia Magnética , Masculino , Síndrome de Marfan/genética , Persona de Mediana EdadRESUMEN
Marfan syndrome, a human disease involving cardiovascular and skeletal apparatuses and ocular and central nervous systems, is associated to mutations in FBN1 gene; heterozygous mutations in TGFBR2 and TGFBR1 genes were found associated to MFS type 2, characterized by the presence of skeletal and cardiovascular major criteria and absence of eye major criterion. We screened the TGFBR1 gene in 46 Marfan patients in whom mutations in FBN1 and TGFBR2 genes were excluded and the analysis of Ex1 was extended to additional 114 Marfan patients and 237 controls. We detected two potentially pathological sequence variants: the TGFBR1 6Ala allele whose frequency was higher in the group of Marfan patients (0.13) than in the controls (0.08) (p=0.013; OR=1.69) and an insertion of 20 nucleotides in the 5'UTR that turned out to be a familial silent rare polymorphism. We hypothesize that TGFBR1 sequence variants may act not only as major, but also as low penetrance alleles of the clinical phenotype in Marfan syndrome.
Asunto(s)
Alelos , Síndrome de Marfan/genética , Penetrancia , Proteínas Serina-Treonina Quinasas/genética , Receptores de Factores de Crecimiento Transformadores beta/genética , Adolescente , Adulto , Secuencia de Bases , Niño , Femenino , Marcadores Genéticos/genética , Variación Genética/genética , Humanos , Masculino , Síndrome de Marfan/diagnóstico , Persona de Mediana Edad , Datos de Secuencia Molecular , Receptor Tipo I de Factor de Crecimiento Transformador beta , Adulto JovenRESUMEN
Homocystinuria is a metabolic disorder associated with defects in genes encoding for methionine metabolism enzymes. Vascular and connective tissue manifestations such as deep venous thrombosis, ectopia lentis and skeletal alterations are the major clinical features. We investigated the clinical manifestations of 5 Italian homocystinuric patients, performed mutation screening analysis on cystationine beta-synthase (CBS) gene and searched for genotype/phenotype correlations. We detected mild cardiovascular and skin connective tissue stigmas in these patients, never reported in homocystinuric patients before. We found 1 novel and 7 known mutations. Our patients carried no other mutation associated with venous thrombosis. Our data stress the importance of extending the clinical investigation for connective tissue manifestations in homocystinuric patients to all the organs/systems involved in Marfan syndrome, also suggesting long term follow-ups for cardiovascular manifestations.
Asunto(s)
Pruebas Genéticas , Homocistinuria/genética , Síndrome de Marfan/genética , Embolia Pulmonar/genética , Trombosis de la Vena/genética , Adulto , Vasos Sanguíneos , Tejido Conectivo , Femenino , Humanos , Italia , Masculino , Adulto JovenRESUMEN
Zac1 is a zinc finger transcription factor that elicits antiproliferative activity and is a potential tumor suppressor gene. Through a detailed spatiotemporal study by in situ hybridization of mouse embryos, we have found that Zac1 transcript is predominantly localized in developing chondrogenic tissue, in addition to the central nervous system as reported elsewhere. Zac1 is also expressed transiently in the myocardium, skeletal muscle, and basal aspect of the stratified embryonic epithelia. During cartilage development, the pattern of Zac1 expression is in close accordance with the distribution of type II collagen mRNA in mesenchymal condensation and prehypertrophic chondrocytes. In mouse ATDC5 cells undergoing in vitro chondrogenesis, the Zac1 mRNA is up-regulated in parallel with genes expressed in precartilage but the Zac1 expression is low when type II collagen mRNA is markedly increased in differentiated cells. Together, these results suggest that Zac1 is a potential regulatory gene involved in chondrogenic differentiation.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Condrogénesis/fisiología , Desarrollo Embrionario y Fetal/fisiología , Factores de Transcripción/metabolismo , Animales , Sistema Nervioso Central/citología , Sistema Nervioso Central/embriología , Epitelio/embriología , Regulación del Desarrollo de la Expresión Génica/fisiología , Genes Supresores de Tumor , Hibridación in Situ , Ratones , Ratones Endogámicos , Músculo Esquelético/citología , Músculo Esquelético/embriología , Miocardio/citología , Miocardio/metabolismo , Especificidad de Órganos , Células Tumorales Cultivadas , Dedos de ZincRESUMEN
AIMS: The aim of this study was to evaluate (1) homocysteinemia and the prevalence of the C677T MTHFR polymorphism in Marfan patients and (2) whether the severity of cardiovascular manifestations is associated with homocysteinemia and/or C677T polymorphism. METHODS AND RESULTS: We studied 107 patients subdivided into three subgroups based on the severity of cardiovascular manifestations: (A) no involvement (n=4); (B) mild involvement (n=45); (C) aortic dilatation or aortic dissection (n=58), and 189 controls. Total homocysteine (tHcy) was significantly higher in subgroup C than in subgroup B. In subgroup C patients with dissection tHcy was higher than in those without dissection. In subgroup C the prevalence of 677T homozygotes was higher, but not significantly, than in the subgroup B. In patients with dissection the prevalence of 677T homozygotes was significantly higher than in those without dissection and than in subgroup B. In the logistic regression analysis, severe cardiovascular manifestations and aortic dissection in Marfan patients were associated with tHcy plasma levels. CONCLUSIONS: Our data indicate an association between the severity of the cardiovascular manifestations, in particular aortic dissection, and elevated tHcy levels. This suggests an important role for tHcy in determining phenotypic variability in Marfan patients and provides further evidence for the association of homocysteinemia with the damage of the vascular system.
Asunto(s)
Enfermedades Cardiovasculares/genética , Hiperhomocisteinemia/genética , Síndrome de Marfan/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Polimorfismo Genético , Adolescente , Adulto , Disección Aórtica/sangre , Disección Aórtica/genética , Aneurisma de la Aorta/sangre , Aneurisma de la Aorta/genética , Enfermedades Cardiovasculares/sangre , Femenino , Homocisteína/sangre , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Fenotipo , Índice de Severidad de la EnfermedadRESUMEN
BACKGROUND: Activated clotting time (ACT) is used to monitor heparin therapy during cardiopulmonary bypass, interventional cardiology, and hemodialysis. Traditionally, ACT is performed by use of the Hemochron system. Recently, a new device, the i-STAT system, has been introduced to measure ACT. The aim of this study was to correlate the performances of these two systems and to compare ACT values with heparin levels. METHODS: One hundred sixty-five samples from 29 patients undergoing cardiopulmonary bypass or hemodialysis were assayed in duplicate with two Hemochron and two i-STAT devices. Heparin levels were determined by anti-factor Xa assay. RESULTS: The Hemochron ACT ranged from 88 to 1,028 s, and the i-STAT ACT ranged from 80 to 786 s. Heparin plasma levels ranged from 0.01 to 10.8 U/mL. Bland-Altman analysis showed a mean difference between the two methods of 24 +/- 101 s. Strong relationships between anti-factor Xa activity and Hemochron ACTs (r2 = 0.69, P < 0.001) and i-STAT ACTs (r2 = 0.79, P < 0.001) were observed. During cardiac surgery, significant correlations were found: Hemochron, r2 = 0.61, P < 0.001 and i-STAT, r2 = 0.74, P < 0.001. During hemodialysis, relationships between anti-factor Xa activity and ACTs were found: Hemochron, r2 = 0.62, P < 0.001 and i-STAT, r2 = 0.55, P < 0.001. CONCLUSIONS: During cardiopulmonary bypass procedure and hemodialysis, i-STAT provides measurements of clotting time quite similar to Hemochron ACT, which were significantly correlated with heparin levels.