A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia.

Aims. To develop a fast and robust DNA-based assay to detect insertions and deletions mutations in exon 34 that encodes the PEST domain of NOTCH1 in order to evaluate patients with chronic lymphocytic leukemia (CLL). Methods. We designed a multiplexed allele-specific polymerase chain reaction (PCR) combined with a fragment analysis assay to detect specifically the mutation c.7544_7545delCT and possibly other insertions and deletions in exon 34 of NOTCH1. Results. We evaluated our assay in peripheral blood samples from two cohorts of patients with CLL. The frequency of NOTCH1 mutations was 8.4% in the first cohort of 71 unselected CLL patients. We then evaluated a second cohort of 26 CLL patients with known cytogenetic abnormalities that were enriched for patients with trisomy 12. NOTCH1 mutations were detected in 43.7% of the patients with trisomy 12. Conclusions. We have developed a fast and robust assay combining allele-specific PCR and fragment analysis able to detect NOTCH1 PEST domain insertions and deletions.

Brazilian experience using high dose sequential chemotherapy followed by autologous hematopoietic stem cell transplantation for malignant lymphomas

OBJECTIVE: To evaluate the use of high-dose sequential chemotherapy in a Brazilian population. METHODS: High-dose cyclophosphamide followed by autologous hematopoietic stem cell transplantation is an effective and feasible therapy for refractory/relapsed lymphomas; this regimen has never before been evaluated in a Brazilian population. All patients (106 with high-grade non-Hodgkin lymphoma and 77 with Hodgkin's lymphoma) submitted to this treatment between 1998 and 2006 were analyzed. Chemotherapy consisted of the sequential administration of high-dose cyclophosphamide (4 or 7 g/m²) and granulocyte-colony stimulating factor (300 µg/day), followed by peripheral blood progenitor cell harvesting, administration of etoposide (2g/m²) and methotrexate (8 g/m² only for Hodgkin's lymphoma) and autologous hematopoietic stem cell transplantation. RESULTS: At diagnosis, non-Hodgkin lymphoma patients had a median age of 45 (range: 8-65) years old, 78 percent had diffuse large B-cell lymphoma and 83 percent had stage III/IV disease. The Hodgkin's lymphoma patients had a median age of 23 (range: 7-68) years old, 64.9 percent had the nodular sclerosis subtype and 65 percent had stage III/IV disease. Nine Hodgkin's lymphoma patients (13 percent) and 10 (9 percent) non-Hodgkin lymphoma patients had some kind of cardiac toxicity. The overall survival, disease-free survival and progression-free survival in Hodgkin's lymphoma were 29 percent, 59 percent and 26 percent, respectively. In non-Hodgkin lymphoma, these values were 40 percent, 49 percent and 31 percent, respectively. High-dose cyclophosphamide-related mortality was 10 percent for Hodgkin's lymphoma and 5 percent for non-Hodgkin lymphoma patients. High-dose cyclophosphamide dosing had no impact on toxicity or survival for both groups. CONCLUSIONS: Despite a greater prevalence of poor prognostic factors, our results are comparable to the literature. The incidence of secondary neoplasias is noteworthy. ...

Brazilian experience using high dose sequential chemotherapy followed by autologous hematopoietic stem cell transplantation for malignant lymphomas.

OBJECTIVE: To evaluate the use of high-dose sequential chemotherapy in a Brazilian population. METHODS: High-dose cyclophosphamide followed by autologous hematopoietic stem cell transplantation is an effective and feasible therapy for refractory/relapsed lymphomas; this regimen has never before been evaluated in a Brazilian population. All patients (106 with high-grade non-Hodgkin lymphoma and 77 with Hodgkin's lymphoma) submitted to this treatment between 1998 and 2006 were analyzed. Chemotherapy consisted of the sequential administration of high-dose cyclophosphamide (4 or 7 g/m(2)) and granulocyte-colony stimulating factor (300 µg/day), followed by peripheral blood progenitor cell harvesting, administration of etoposide (2g/m(2)) and methotrexate (8 g/m(2) only for Hodgkin's lymphoma) and autologous hematopoietic stem cell transplantation. RESULTS: At diagnosis, non-Hodgkin lymphoma patients had a median age of 45 (range: 8-65) years old, 78% had diffuse large B-cell lymphoma and 83% had stage III/IV disease. The Hodgkin's lymphoma patients had a median age of 23 (range: 7-68) years old, 64.9% had the nodular sclerosis subtype and 65% had stage III/IV disease. Nine Hodgkin's lymphoma patients (13%) and 10 (9%) non-Hodgkin lymphoma patients had some kind of cardiac toxicity. The overall survival, disease-free survival and progression-free survival in Hodgkin's lymphoma were 29%, 59% and 26%, respectively. In non-Hodgkin lymphoma, these values were 40%, 49% and 31%, respectively. High-dose cyclophosphamide-related mortality was 10% for Hodgkin's lymphoma and 5% for non-Hodgkin lymphoma patients. High-dose cyclophosphamide dosing had no impact on toxicity or survival for both groups. CONCLUSIONS: Despite a greater prevalence of poor prognostic factors, our results are comparable to the literature. The incidence of secondary neoplasias is noteworthy. Our study suggests that this approach is efficient and feasible, regardless of toxicity-related mortality.

Incidence of aplastic anemia and agranulocytosis in Latin America – The LATIN study

CONTEXTO E OBJETIVO: A anemia aplástica e a agranulocitose são doenças raras, entretanto freqüentemente letais. Muitas vezes são causadas por medicações e outras exposições ambientais. A incidência dessas doenças parece variar consideravelmente entre diferentes regiões geográficas, e poucos dados sobre a incidência são disponíveis para os países da América Latina. O objetivo deste trabalho é determinar a incidência de anemia aplástica e agranulocitose no Brasil. TIPO DE ESTUDO E LOCAL: Estudo de incidência. Sete centros participaram da fase piloto do estudo representando as cinco regiões brasileiras. MÉTODOS: Cada centro realizou busca ativa por novos casos em uma região definida, por meio de contatos regulares com todos os hematologistas, principais laboratórios clínicos e clínicos de hospitais de sua região. RESULTADOS: Foram identificados 74 casos de anemia aplástica e 16 casos de agranulocitose. A mediana de idade dos pacientes com agranulocitose foi de 31 anos (intervalo inter-quartil – IIQ 12,5 – 48,2), 32,2% eram do sexo masculino e 81,2% eram da raça branca. A mediana de idade dos pacientes com anemia aplástica foi de 21 anos (IIQ 15,0-35,2), 62,2% eram do sexo masculino, 50,0% da raça branca e 39,2% da raça parda. A incidência de agranulocitose foi estimada em 0,5 casos/milhão de habitantes/ano, variando de 0,0 a 1,1 caso/milhão de habitantes/ano entre as diferentes regiões brasileiras. A incidência de anemia aplástica foi de 2,7 casos/milhão de habitantes/ano, variando de 1,1 a 7,1 casos/milhão de habitantes/ano entre as diferentes regiões. CONCLUSÕES: A anemia aplástica e a agranulocitose são doenças raras no Brasil. Entretanto existe considerável variabilidade na incidência destas doenças entre as diferentes regiões brasileiras.