RESUMO
Seventy-nine patients with AML in CR1 received allo-SCT between May 2006 and May 2011, and the prognostic impact of FMS-like tyrosine kinase 3/internal tandem duplication (FLT3/ITD) mutation was evaluated in the context of other clinical prognostic factors. Patients with FLT3/ITD + AML had significantly inferior DFS (2-year DFS: 19% vs 64%, P = 0.0027), increased risk of relapse (1-year: 59% vs 19%, P = 0.01), and a trend towards decreased OS (P = 0.08) compared with patients without FLT3/ITD. Multivariate analysis confirmed FLT3/ITD + independently predicted a shorter DFS (HR, 3.0; 95% CI), 1.4-6.5; P = 0.01) and increased risk of relapse (HR, 4.9; 95% CI, 2.0-12.3, P = 0.01). Time to relapse in patients with FLT3/ITD + was short with 100-day cumulative risk of 45% (95% CI, 33-57). Our data suggest that the poor prognostic implication of FLT3/ITD positivity remains even after early allo-SCT in patients with FLT3/ITD + AML, and patients remain at high risk of early relapse. FLT3/ITD positivity also outweighs other conventional prognostic markers in predicting relapse.
Assuntos
Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/cirurgia , Transplante de Células-Tronco/métodos , Tirosina Quinase 3 Semelhante a fms/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Terapia Combinada , Feminino , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/enzimologia , Masculino , Pessoa de Meia-Idade , Mutação , Prognóstico , Análise de Sobrevida , Sequências de Repetição em Tandem , Transplante Homólogo , Resultado do Tratamento , Adulto JovemRESUMO
The promise of "personalized medicine" guided by an understanding of each individual's genome has been fostered by increasingly powerful and economical methods to acquire clinically relevant information. We describe the operational implementation of prospective genotyping linked to an advanced clinical decision-support system to guide individualized health care in a large academic health center. This approach to personalized medicine entails engagement between patient and health-care provider, identification of relevant genetic variations for implementation, assay reliability, point-of-care decision support, and necessary institutional investments. In one year, approximately 3,000 patients, most of whom were scheduled for cardiac catheterization, were genotyped on a multiplexed platform that included genotyping for CYP2C19 variants that modulate response to the widely used antiplatelet drug clopidogrel. These data are deposited into the electronic medical record (EMR), and point-of-care decision support is deployed when clopidogrel is prescribed for those with variant genotypes. The establishment of programs such as this is a first step toward implementing and evaluating strategies for personalized medicine.
Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , Cateterismo Cardíaco/efeitos dos fármacos , Farmacogenética , Medicina de Precisão , Ticlopidina/análogos & derivados , Cateterismo Cardíaco/métodos , Clopidogrel , Desenho Assistido por Computador , Citocromo P-450 CYP2C19 , Sistemas de Apoio a Decisões Clínicas , Variação Genética , Técnicas de Genotipagem/métodos , Humanos , Seleção de Pacientes , Farmacogenética/métodos , Farmacogenética/tendências , Inibidores da Agregação Plaquetária/uso terapêutico , Medicina de Precisão/métodos , Medicina de Precisão/tendências , Ticlopidina/uso terapêuticoRESUMO
RATIONALE: Mounting data suggest that immune cell abnormalities participate in the pathogenesis of pulmonary arterial hypertension (PAH). OBJECTIVE: To determine whether the T lymphocyte subset composition in the systemic circulation and peripheral lung is altered in PAH. METHODS: Flow cytometric analyses were performed to determine the phenotypic profile of peripheral blood lymphocytes in idiopathic PAH (IPAH) patients (n=18) and healthy controls (n=17). Immunocytochemical analyses of lymphocytes and T cell subsets were used to examine lung tissue from PAH patients (n=11) and controls (n=11). MEASUREMENTS AND MAIN RESULTS: IPAH patients have abnormal CD8+ T lymphocyte subsets, with a significant increase in CD45RA+ CCR7- peripheral cytotoxic effector-memory cells (p=0.02) and reduction of CD45RA+ CCR7+ naive CD8+ cells versus controls (p=0.001). Further, IPAH patients have a higher proportion of circulating regulatory T cells (T(reg)) and 4-fold increases in the number of CD3+ and CD8+ cells in the peripheral lung compared with controls (p<0.01). CONCLUSIONS: Alterations in circulating T cell subsets, particularly CD8+ T lymphocytes and CD4+ T(reg), in patients with PAH suggest that a dysfunctional immune system contributes to disease pathogenesis. A preponderance of CD3+ and CD8+ T lymphocytes in the peripheral lung of PAH patients supports this concept.
Assuntos
Hipertensão Pulmonar/imunologia , Pulmão/imunologia , Subpopulações de Linfócitos T/imunologia , Adulto , Feminino , Citometria de Fluxo , Humanos , Memória Imunológica/imunologia , Pulmão/química , Masculino , Pessoa de Meia-IdadeAssuntos
Sangue , Medula Óssea , Transplante de Células-Tronco , Quimeras de Transplante , Adolescente , Adulto , Criança , Feminino , Neoplasias Hematológicas/terapia , Humanos , Masculino , Repetições de Microssatélites , Pessoa de Meia-Idade , Estudos Retrospectivos , Sensibilidade e Especificidade , Transplante Homólogo , Resultado do TratamentoRESUMO
Previous reports have suggested that expansion of the CGG repeat located in the fragile X mental retardation 1 (FMR1) gene might be responsible for a significant number of patients with the multiple system atrophy (MSA) phenotype. Analysis of 65 MSA patients found only 4.6% displayed CGG expansions in the suspected range. This is similar to the frequency reported in the normal population, suggesting that this expansion does not play a major role in the MSA phenotype.
Assuntos
Atrofia de Múltiplos Sistemas/genética , Proteínas do Tecido Nervoso/genética , Proteínas de Ligação a RNA/genética , Expansão das Repetições de Trinucleotídeos , Idoso , Idoso de 80 Anos ou mais , Feminino , Proteína do X Frágil da Deficiência Intelectual , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , RNA Mensageiro/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fatores SexuaisRESUMO
Described is a patient with concurrent discrete gliomas: a pleomorphic xanthoastrocytoma with anaplastic features and an anaplastic oligoastrocytoma. The distinct and morphologically dissimilar tumors demonstrated similar genetic abnormalities by loss of heterozygosity and comparative genome hybridization. Clonality and proteomic analyses highlighted an independent origin for the two tumors. Proteomic methods may prove useful in cases where the differential diagnosis and pathogenetic origin of tumors are uncertain, as well as more globally for its ability to provide insight into specific expression of proteins that may serve as unique markers of tumorigenesis or as novel targets of therapy.
Assuntos
Neoplasias Encefálicas/química , Neoplasias Encefálicas/genética , Glioma/química , Glioma/genética , Proteínas de Neoplasias/análise , Neoplasias Primárias Múltiplas/química , Neoplasias Primárias Múltiplas/genética , Proteoma/análise , Neoplasias Encefálicas/patologia , Mapeamento Cromossômico , Eletroforese em Gel Bidimensional , Feminino , Glioma/patologia , Humanos , Perda de Heterozigosidade , Pessoa de Meia-Idade , Neoplasias Primárias Múltiplas/patologia , Hibridização de Ácido NucleicoAssuntos
Rearranjo Gênico da Cadeia gama dos Receptores de Antígenos dos Linfócitos T/genética , Erupções Liquenoides/imunologia , Púrpura/imunologia , Dermatopatias/imunologia , Linfócitos T/imunologia , Pré-Escolar , Células Clonais , Feminino , Humanos , Lactente , Masculino , Reação em Cadeia da PolimeraseRESUMO
Hepatosplenic gammadelta T-cell lymphoma is a distinct entity, characterized by occurrence in young adult males with hepatosplenomegaly, B-symptoms, peripheral blood cytopenias, and no lymphadenopathy; lymphomatous infiltrates in the splenic red pulp, hepatic sinusoids, and bone marrow sinuses; T-cell receptor (TCR) gammadelta chains and a cytotoxic T-cell phenotype; isochromosome 7q; and an aggressive clinical course. In comparison, this study describes the clinicopathologic features of 14 hepatosplenic T-cell lymphomas expressing TCR alphabeta chains. They occurred in 11 women and 3 men with a median age of 36 years. Clinical presentation was similar to that described previously for hepatosplenic gammadelta T-cell lymphomas, except for the female preponderance and age distribution (5 patients younger than 13 years of age and 5 patients older than 50 years of age). Disease distribution was primarily in the splenic red pulp and hepatic sinusoids, although liver infiltrates were largely periportal in four cases. Bone marrow involvement, observed in eight patients, was usually interstitial and/or within the sinuses. Lymph nodes were involved in five patients, although lymphadenopathy was demonstrable in only two. Ten cases were composed of intermediate-size tumor cells with round/oval nuclei, slightly dispersed chromatin, inconspicuous nucleoli, and scant to moderate amounts of cytoplasm. Four lymphomas contained primarily large cells with irregular nuclei, dispersed chromatin, discernible nucleoli, and moderate to abundant cytoplasm. Tumor cells in all 14 lymphomas were cytotoxic alphabeta T-cells; 13 co-expressed natural killer cell-associated antigens and showed T-cell clonality. Three lymphomas were associated with Epstein-Barr virus. Two of four cases had an isochromosome 7q. Eleven patients are dead, eight within a year of diagnosis, and two patients have maintained complete remissions after combination chemotherapy. These data show that hepatosplenic T-cell lymphomas include an alphabeta-subtype. This group, along with the previously recognized gammadelta group, should be recognized as phenotypically heterogeneous subtypes of the same disease entity.
Assuntos
Neoplasias Hepáticas/patologia , Linfoma de Células T/patologia , Receptores de Antígenos de Linfócitos T alfa-beta/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Neoplasias Esplênicas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Medula Óssea/patologia , Criança , Pré-Escolar , DNA de Neoplasias/análise , Feminino , Rearranjo Gênico da Cadeia gama dos Receptores de Antígenos dos Linfócitos T/genética , Humanos , Lactente , Recém-Nascido , Cariotipagem , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Linfonodos/patologia , Linfoma de Células T/classificação , Linfoma de Células T/genética , Linfoma de Células T/metabolismo , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Antígenos de Linfócitos T gama-delta/genética , Neoplasias Esplênicas/genética , Neoplasias Esplênicas/metabolismoRESUMO
The Mohr-Tranebjaerg syndrome (MIM 304700) and the Jensen syndrome (MIM 311150) were previously reported as separate X-linked recessive deafness syndromes associated with progressive visual deterioration, dystonia, dementia, and psychiatric abnormalities. In the most extensively studied Norwegian family, the Mohr-Tranebjaerg syndrome was reported to be caused by a one-basepair deletion (151delT) in the deafness/dystonia peptide (DDP) gene at Xq22. This gene has been renamed TIMM8a. We identified a stop mutation (E24X) in the TIMM8a gene segregating with the disease in the original Danish family with the Jensen syndrome, which confirms that the two disorders are allelic conditions. We also report abnormal VEP examinations and neuropathological abnormalities in affected males from the two unrelated families with different mutations. The findings included neuronal cell loss in the optic nerve, retina, striate cortex, basal ganglia, and dorsal roots of the spinal cord. The demonstration of mitochondrial abnormalities in skeletal muscle biopsies in some patients is compatible with the suggestion from recent research that the TIMM8a protein is the human counterpart of an intermembrane mitochondrial transport protein, Tim8p, recently characterized in yeast. The clinical and neuropathological abnormalities associated with mutations in the TIMM8a gene support that this X-linked deafness-dystonia-optic neuropathy syndrome is an example of progressive neurodegeneration due to mutations in a nuclear gene necessary for some, yet unknown mitochondrial transport function. We recommend sequencing the TIMM8a gene, thorough ophthalmological examination, and measuring visual evoked potentials in clinically suspected male patients with either progressive hearing impairment, dystonia, or visual disability in order to establish an early diagnosis and provide appropriate genetic counselling.
Assuntos
Surdez/genética , Distonia/genética , Doenças Mitocondriais/genética , Mutação/genética , Doenças do Nervo Óptico/genética , Proteínas/genética , Córtex Visual/patologia , Cromossomo X/genética , Adolescente , Adulto , Idoso , Morte Celular , Criança , Análise Mutacional de DNA , Surdez/patologia , Distonia/patologia , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Potenciais Evocados Visuais , Feminino , Genes Recessivos , Ligação Genética , Humanos , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-Idade , Doenças Mitocondriais/fisiopatologia , Neurônios/patologia , Doenças do Nervo Óptico/patologia , Linhagem , Fosfopiruvato Hidratase/metabolismo , Reação em Cadeia da Polimerase , SíndromeRESUMO
CAG repeat number in the androgen receptor (AR) has been associated with decreased prostate cancer risk, and AR expression has been found in female breast cancer, often associated with apocrine differentiation. Because trinucleotide expansion can alter gene expression and protein function, we hypothesized that it might occur in breast neoplasms. We used a repeat expansion detection technique to determine CAG repeat lengths in DNA from breast biopsies. Three lesion types were microdissected: fibroadenoma (48 cases), ductal carcinoma in situ (DCIS, 24 cases), and invasive mammary carcinoma (18 cases). The maximum number of CAG repeats in either allele of each patient in these three groups was compared. Microsatellite repeat lengths in DCIS were longer than in fibroadenomas or invasive carcinomas (P= 0.017 comparing DCIS vs invasive carcinomas). Two cases of apocrine DCIS had very long repeat lengths, both exhibiting microsatellite lengths at the longest range of normal (32 and 33). Inherited differences in AR CAG length might influence the transition from DCIS to invasive breast cancer, perhaps by modulating function of AR in breast tissue. AR microsatellite polymorphisms could influence cellular differentiation in DCIS lesions, promoting formation of the apocrine subtype in the presence of longer CAG repeats.
RESUMO
We report a case of severe haemophilia A (<1% factor VIII level) in a female resulting from an interesting and improbable combination of events. The patient inherited a factor VIII intron 22 inversion from her carrier mother, as well as a second factor VIII inversion involving intron 22 that arose de novo on her paternally derived X chromosome. In addition, the patient's paternally derived X chromosome had been preferentially inactivated in 95+% of her somatic cells. The patient's mother, who was clinically unaffected, carried an intron 22 inversion as well and also showed nonrandom X-inactivation. The patient's mother had a brother with severe haemophilia A. It is therefore likely that the mother's inversion was on her maternally derived X chromosome. Since she was unaffected, it is likely that her inversion-bearing X was the one that was preferentially inactivated.
Assuntos
Hemofilia A/genética , Aberrações dos Cromossomos Sexuais/genética , Southern Blotting , Inversão Cromossômica , Metilação de DNA , Fator VIII/genética , Saúde da Família , Feminino , Inativação Gênica , Heterozigoto , Humanos , Lactente , Cromossomo X/genéticaRESUMO
Technological advances in molecular biology, coupled with the Human Genome Project, has led to the isolation and characterization of thousands of human genes. Subsequently, many of these discoveries have been promptly translated into clinical assays by DNA laboratories for immediate patient evaluation and management. Since a variety of mutation detection techniques exist, the technique most appropriate for clinical testing of a particular disease is determined by: both the type and number of different mutations associated with the disease; the frequency of referrals; and the required turn-around time for appropriate patient management. This review discusses some of the more commonly inherited diseases for which molecular testing is available. It describes and illustrates the techniques used for direct mutation analysis of expanded trinucleotide repeats, point mutations, deletions, gene rearrangements, uniparental disomy, and linkage analysis.
Assuntos
Doenças Genéticas Inatas/diagnóstico , Técnicas Genéticas , Testes Genéticos/métodos , Mapeamento Cromossômico , Análise Mutacional de DNA , Bases de Dados Factuais , Feminino , Deleção de Genes , Ligação Genética , Genoma Humano , Humanos , Masculino , Linhagem , Mutação PuntualRESUMO
The cell of origin of parafollicular (monocytoid) B cell lymphoma (PBCL), splenic marginal zone lymphoma (SMZL), and hairy cell leukemia (HCL) is controversial. To better understand the relationship between these low-grade B-cell neoplasms, we analyzed the nucleotide sequences of the rearranged immunoglobulin heavy (IgH) chain variable (V) region of the clonal population of cells in five cases of PBCL, four cases of SMZL, and seven cases of HCL to determine whether these neoplasms could be differentiated by the degree of somatic mutation in the IgH V gene or by the IgH V gene family usage. DNA was extracted from diagnostic material and clonality confirmed by PCR. The DNA was reamplified using V heavy chain family specific primers, and the amplicons were sequenced. Sequences were compared with germline IgH V gene sequences, and base changes were determined to be silent or to represent amino acid replacements by using three different methods. Four of five (80%) cases of PBCL, three of four (75%) cases of SMZL, and three of seven (43%) cases of HCL showed evidence of antigen selection, suggesting that these neoplasms involved clonal expansions of postgerminal center memory lymphocytes. Only SMZL showed a preferential usage of V(H)1 family genes.
Assuntos
Cadeias Pesadas de Imunoglobulinas/genética , Região Variável de Imunoglobulina/genética , Leucemia de Células Pilosas/imunologia , Linfoma de Células B/imunologia , Neoplasias Esplênicas/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Feminino , Rearranjo Gênico , Humanos , Imuno-Histoquímica , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , MutaçãoRESUMO
Microsatellite instability (MSI) is a recently described type of genetic alteration resulting from defects in the DNA mismatch repair genes that appears to play an integral role in neoplastic transformation. MSI has been described in a wide variety of malignancies; however, data regarding the role of MSI in the pathogenesis of non-Hodgkin's lymphoma (NHL) are limited. MSI appears to be important in some T-cell lymphomas, including ALCL arising in immunocompromised patients. In addition, MSI has recently been identified in CD30+ cutaneous lymphoproliferative processes and lymphoblastic lymphoma. In this study, we have analyzed five well-characterized cases of systemic T-cell ALCL arising in immunocompetent patients for the presence of MSI. Genomic DNA isolated from paired normal and tumor tissue was analyzed at seven microsatellite loci by polymerase chain reaction. We were unable to identify MSI or loss of heterozygosity (LOH) in our cases, suggesting that abnormalities in the DNA mismatch repair system do not play a major role in the pathogenesis of most systemic ALCL. Our data provide additional molecular evidence that the various subgroups of lymphoma with ALCL morphology are biologically distinct processes.
Assuntos
Linfoma Anaplásico de Células Grandes/genética , Adolescente , Adulto , Idoso , Feminino , Humanos , Imunocompetência , Perda de Heterozigosidade , Linfoma Anaplásico de Células Grandes/patologia , Masculino , Repetições de MicrossatélitesRESUMO
Cystic fibrosis (CF) is an autosomal recessive disease caused by at least 750 different mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The frequency of the most common mutation (DF508) in Brazilian patients of European origin is 47%. To determine the frequency of 4 other common CF mutations (G542X, G551D, R553X, and N1303K) in Brazilian patients of European origin, we used direct polymerase chain reaction (PCR) amplification of DNA obtained from dried blood spots on Guthrie cards. The DNA came from 247 non-DF508 chromosomes from 172 Brazilian CF patients ascertained from 5 different states of Brazil. The results show that the 4 mutations account for 17% of the non-DF508 alleles and only 9% of the total number of Brazilian CF alleles. Overall, the frequency of each mutation is different from northern European and North American populations but similar to southern European populations, mainly the Italian and Spanish populations. When Brazilian patients of European origin are grouped according to state of birth, the frequencies of the mutations are significantly different between southern and southeastern states of Brazil. Therefore there are serious implication for risk assessment of DNA-based tests in heterogeneous populations such as Brazilians. Further studies are needed to identify the remaining 44% of CF mutations for the different populations and regions of Brazil.
Assuntos
Fibrose Cística/epidemiologia , Fibrose Cística/genética , DNA Satélite/análise , Frequência do Gene , Heterogeneidade Genética , Mutação/genética , Adolescente , Adulto , Brasil/epidemiologia , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Europa (Continente)/etnologia , Humanos , Lactente , Masculino , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Estudos de Amostragem , Estudos SoroepidemiológicosRESUMO
We describe clinical and pathologic features of a patient with fatal familial insomnia (FFI) whose prion (PrP) genotype is D178N coupled with methionine at codon 129 on his mutant allele and valine at codon 129 on his normal allele. A cousin (genetic half brother) with identical PrP genotypes exhibited strikingly different clinical and pathologic changes. Comparison of these cousins shows the phenotypic heterogeneity of FFI and suggests that the phenotypic expression of D178N is influenced by multiple factors.
Assuntos
Saúde da Família , Heterogeneidade Genética , Mutação Puntual , Doenças Priônicas/genética , Doenças Priônicas/patologia , Adulto , Atrofia , Genótipo , Humanos , Masculino , Degeneração Neural/genética , Degeneração Neural/patologia , Núcleo Familiar , Príons/genética , Tálamo/patologiaRESUMO
Fibroadenoma is a common cause of benign breast masses in young women. These women have a slightly increased risk of subsequent breast cancer, particularly if their tumors have complex histologic patterns. We assessed monoclonality in fibroadenomas and correlated the results with histologic analysis. We performed a clonal analysis of 52 fibroadenomas from 43 patients using X-chromosome inactivation studies. The cases included fibroadenomas with complex and simple histology. Areas examined were predominantly stroma but epithelium was also present. DNA was isolated from paraffin-embedded tissue and was subjected to polymerase chain reaction amplification of the human androgen receptor gene with and without predigestion of the DNA with Hha 1. If a monoclonal process was identified, the epithelial and stromal components were subsequently microdissected and reanalyzed. 36/43 (83.7%) women were heterozygous. We studied 45 tumors in these 36 informative women. 1/20 (5% complex fibroadenomas and 1/25 (4%) simple fibroadenomas were monoclonal. The epithelial component of both monoclonal fibroadenomas was polyclonal. The one monoclonal simple fibroadenoma was also the only one with mixed features to contain a phyllodes component. In this case, monoclonality was found in the stroma of both the fibroadenoma and phyllodes regions. Monoclonality has been previously associated with phyllodes phenotype, but not with fibroadenomas, except for 3 fibroadenomas that recurred as phyllodes tumor. We report that monoclonality may also be seen occasionally in complex fibroadenomas, and was found in a tumors with mixed fibroadenoma/phyllodes features without clinical recurrence for 4 years.
Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Mecanismo Genético de Compensação de Dose , Fibroadenoma/genética , Fibroadenoma/patologia , Tumor Filoide/genética , Tumor Filoide/patologia , Primers do DNA , Feminino , Humanos , Reação em Cadeia da PolimeraseRESUMO
Chagas' disease is caused by the hemoflagellate protozoan Trypanosoma cruzi. The most common, serious manifestation of Chagas' disease is a progressive inflammatory cardiomyopathy, which occurs decades after primary infection. The inability to consistently demonstrate T. cruzi by histologic techniques in inflammatory cardiac lesions has suggested that the parasites' persistence may not be required for the pathology of the chronic phase. In this report we further analyze the persistence and localization of T. cruzi DNA in the hearts of seven patients with chronic chagasic cardiomyopathy, along with four indeterminate patients and seven control patients seronegative for T. cruzi infection. In the seven patients with chronic chagasic cardiomyopathy, we extracted DNA from selected inflammatory foci-positive (IFP) and inflammatory foci-negative (IFN) areas of' hematoxylin and eosin-stained cardiac tissue. We then used polymerase chain reaction methodology to amplify three different T. cruzi sequences (a minicircle sequence [MCS], a satellite repetitive sequence [RS], and, a low copy number sequence within the gene coding for a flagellar protein [FPS]). The MCS was detected in approximately 100% of both the IFP and IFN areas analyzed. The RS was detected in 37.5% and 23% of the IFP and IFN areas, respectively (difference not statistically significant; P > 0.10, degrees of freedom = 1, G test of independence = 1.9522). The FPS was rarely detected (2%), and was only present in DNA extracted from IFP areas. The MCS was also detected in most indeterminate cases (none of whom had inflammatory lesions) although with a markedly diminished amplification signal relative to cardiomyopathy cases. The MCS was not amplified from the cardiac tissues from seronegative controls. These results suggest that the quantity of T. cruzi DNA persisting in hearts of patients with Chagas' disease correlates with cardiomyopathy, but may not be preferentially associated with inflammatory foci.
Assuntos
Cardiomiopatia Chagásica/parasitologia , DNA de Protozoário/análise , Coração/parasitologia , Reação em Cadeia da Polimerase , Trypanosoma cruzi/isolamento & purificação , Animais , Humanos , Trypanosoma cruzi/genéticaRESUMO
BACKGROUND: The APOE genotype has been shown to influence the risk of developing sporadic and familial AD. This effect is isoform-dependent, the APOE epsilon4 allele increasing susceptibility and the APOE epsilon2 allele providing protection. Amyloid formation is an important part of the pathogenesis in AD as well as in spongiform encephalopathies; apoE deposition in amyloid plaques has been documented in both conditions. METHODS: We examined the frequency of the APOE alleles in patients with various forms of transmissible spongiform encephalopathies, or prion diseases, including sporadic and iatrogenic Creutzfeldt-Jakob disease; familial Creutzfeldt-Jakob disease associated with PRNP 178N/129V and 200K/129M point mutations and a 24-nucleotide repeat expansion; fatal familial insomnia caused by the 178N/129M mutation; Gerstmann-Sträussler-Scheinker disease associated with 102L/129M mutation; and kuru. RESULTS: None of the groups we studied had a significant excess of APOE epsilon4 allele when compared with appropriate controls. CONCLUSION: Our results do not support the contention that the APOE epsilon4 allele is a risk factor for developing Creutzfeldt-Jakob disease or related disorders.
Assuntos
Amiloidose/genética , Apolipoproteínas E/genética , Doenças Priônicas/genética , Alelos , Síndrome de Creutzfeldt-Jakob/genética , Genótipo , Doença de Gerstmann-Straussler-Scheinker/genética , Humanos , Doença Iatrogênica , Kuru/genética , Mutação , Periodicidade , Fatores de RiscoRESUMO
The rate of parvovirus B19 (PV) infection in cases of "idiopathic" nonimmune hydrops fetalis (NIHF) is reported to be approximately 16% with polymerase chain reaction (PCR)-based methods. Antibodies for use in paraffin-embedded tissue have not been systematically compared with PCR or with the presence of inclusions at varying gestational ages. All autopsy cases of NIHF and those with effusions of multiple serous membranes examined between 1991 and 1996 (n = 29) were evaluated for the presence of PV DNA by PCR analysis of paraffin-embedded liver tissue. PCR-positive cases and "idiopathic" cases were examined for the presence of inclusions in routine histological sections and for PV protein using a monoclonal antibody (NovoCastra R92F6). Among the four clinically idiopathic cases, one (25%) was positive for PV using PCR. The three negative idiopathic cases had no inclusions and were negative for PV by PCR and immunohistochemistry (IHC); all were third-trimester gestations (28, 31, and 32 weeks). Identifiable risk factors for NIHF other than PV in the remaining 25 cases included cystic hygroma, seven (three 45,X; two 46,XX; two no growth); complex cardiac anomaly, six; infection, three (two CMV, one chlamydia); twin-twin transfusion, two; lymphangiectasia, two; diaphragmatic hernia, tracheal atresia, trisomy 21, congenital cystic adenomatoid malformation, one each. One of these nonidiopathic cases, a fetus with cystic hygroma and a 45,X karyotype, was positive for PV DNA only on the blot, consistent with a low titer; no inclusions were present, and IHC was negative in multiple organs in this instance. One of four (25%) cases of idiopathic NIHF cases contained PV DNA by PCR analysis; there were abundant inclusions in multiple organs, and IHC was strongly positive as well. Of 25 cases of nonidiopathic NIHF, one (4%) was also positive for PV DNA by PCR. PV protein was detected by IHC only in the presence of inclusions; IHC thus may be useful for highlighting sparse inclusions. No second-trimester case of NIHF was unexplained. Late (third-trimester) cases of "idiopathic" NIHF are likely to be negative by all methods, either because they are not attributable to PV infection or because PV protein and DNA are below detectable levels or are no longer present. Maternal serology for PV and TORCH agents may be the best method for investigating third-trimester losses to otherwise unexplained NIHF.