Functional properties and potential application of ethanol tolerant ß-glucosidases from Pichia ofunaensis and Trichosporon multisporum yeasts.

ß-Glucosidases have been extensively investigated to integrate the enzyme complex for cellulose fiber saccharification and for improving the aroma of wine. To produce these enzymes, greater attention has been given to filamentous fungi and bacteria, and few investigations have targeted the potential applications of enzymes secreted by yeasts. Addressing this issue, in this study, ß-glucosidases were produced by the Pichia ofunaensis and Trichosporon multisporum yeasts, via solid state fermentation with wheat bran as a substrate. When using p-Nitrophenyl ß-d-glucopyranoside (pNPG) as an enzyme substrate, maximum ß-glucosidase activities were detected at pH 5.5-6.0 and 50-60 °C for P. ofunaensis, and pH 5-6 and 55 °C for T. multisporum. Both enzymes were able to hydrolyze cellobiose and exhibited stability over a wide range of pH (3.5-9.0) for 24 h at 4 °C, thermostability up to 50 °C for 1 h and tolerance to 10 mM phenolic compounds. Negative modulation on enzyme activity was observed in the presence of Cu2+, Fe3+, Zn2+, Al3+ and Hg2+, while both ß-glucosidases were tolerant to 30% methanol, isopropanol and acetone. In the presence of ethanol and glucose, enzymes from P. ofunaensis were the more active and stable of the two. These enzymes, especially the P. ofunaensis ß-glucosidases, could be tested in enology for improving the aroma of wine and for integrating a cellulolytic complex to produce 2G ethanol.

A Collagenolytic Aspartic Protease from Thermomucor indicae-seudaticae Expressed in Escherichia coli and Pichia pastoris.

Proteases are produced by the most diverse microorganisms and have a wide spectrum of applications. However, the use of wild microorganisms, mainly fungi, for enzyme production has some drawbacks. They are subject to physiological instability due to metabolic adaptations, causing complications and impairments in the production process. Thus, the objective of this work was to promote the heterologous expression of a collagenolytic aspartic protease (ProTiN31) from Thermomucor indicae seudaticae in Escherichia coli and Pichia pastoris. The pET_28a (+) and pPICZαA vectors were synthesized containing the gene of the enzyme and transformed into E. coli and P. pastoris, respectively. The recombinant enzymes produced by E. coli and P. pastoris showed maximum activity at pH 5.0 and 50 °C, and pH 5.0 and 60 °C, respectively. The enzyme produced by P. pastoris showed better thermostability when compared to that produced by E. coli. Both enzymes were stable at pH 6.0 and 6.5 for 24 h at 4 °C, and sensitive to pepstatin A, ß-mercaptoethanol, and Hg2+. Comparing the commercial collagen hydrolysate (Artrogen duo/Brazil) and gelatin degradation using protease from P. pastoris, they showed similar peptide profiles. There are its potential applications in a wide array of industrial sectors that use collagenolytic enzymes.

Shorter Maintenance Therapy in Childhood Acute Lymphoblastic Leukemia: The Experience of the Prospective, Randomized Brazilian GBTLI ALL-93 Protocol.

AIM: Maintenance therapy is an important phase of the childhood ALL treatment, requiring 2-year long therapy adherence of the patients and families. Weekly methotrexate with daily 6-mercaptopurine (6MP) constitutes the backbone of maintenance therapy. Reduction in the maintenance therapy could overweight problems related with poverty of children with ALL living in limited-income countries (LIC). OBJECTIVE: To compare, prospectively, the EFS rates of children with ALL treated according to two maintenance regimens: 18 vs. 24 months duration. MATERIALS AND METHODS: From October 1993 to September 1999, 867 consecutive untreated ALL patients <18 years of age were treated according to the Brazilian Cooperative Group for Childhood ALL Treatment (GBTLI) ALL-93 protocol. Risk classification was based exclusively on patient's age and leukocyte count (NCI risk group) and clinical extra medullary involvement of the disease. Data were analyzed by the intention-to-treat approach. RESULTS: Fourteen patients (1.6%) were excluded: wrong diagnosis (n = 7) and previous corticosteroid (n = 7). Of the 853 eligible patients, 421 were randomly allocated, at study enrollment, to receive 18-month (group 1) and 432 to receive 24-month (group 2) maintenance therapy. Complete remission rate was achieved in 96% of the patients (817/853). Twenty-eight patients (3.4%) died during the induction phase. Thirty-four patients (4.0%) were lost to follow-up. The overall EFS was 66.1 ± 1.7% at 15 years. No difference was seen according to maintenance: EFS15y was 65.8 ± 2.3% (group 1) and 66.3 ± 2.3% (group 2; p = 0.79). No difference between regimens was detected after stratifying the analyses according to factors associated with adverse prognosis in this study (age group <1 year or >10 years and high WBC at diagnosis). Overall death in remission rate was 6.85% (56 patients). Deaths during maintenance were 13 in group 1 and 12 in group 2, all due to infection. Over 15 years of follow-up, two patients both from group 2 presented a second malignancy (Hodgkin's disease and thyroid carcinoma) after 8.3 and 11 years off therapy, respectively. CONCLUSION: Six-month reduction of maintenance therapy in ALL children treated according to the GBTLI ALL-93 protocol provided the same overall outcome as 2-year duration regimen.

Carboplatin in the treatment of Ewing sarcoma: Results of the first Brazilian collaborative study group for Ewing sarcoma family tumors-EWING1.

BACKGROUND: Large cooperative group studies have shown the efficacy of risk-adapted treatment for Ewing sarcoma. However, validation and local adaptation by National cooperative groups is needed. A multicenter protocol to determine the efficacy and safety of a risk-adapted intensive regimen was developed by the Brazilian cooperative group. PROCEDURE: Patients <30 years old with Ewing sarcoma were eligible. Induction chemotherapy consisted of two cycles of ICE (ifosfamide, carboplatin, and etoposide) followed by two cycles of VDC (vincristine, doxorubicin, and cyclophosphamide), followed by local control. Patients with low risk (LR) disease (localized resectable with normal LDH) received 10 additional alternating courses of IE with VDC. For patients with high-risk (HR) disease (unresectable, pelvic, metastatic, or high LDH), two additional cycles of ICE were given. RESULTS: One-hundred seventy five patients (39% metastatic) were enrolled. Fifty-two patients (29.7%) were LR and 123 (70.3%) were HR. Overall response rate at end of induction was 27.4%. Five-year event-free survival (EFS) and overall survival (OS) estimates were 51.4% and 54.4%, respectively. Patients with localized disease had better outcomes than patients with metastases (5-year EFS 67.9% vs. 25.5%, and 5-year OS 70.3% vs. 29.1%, respectively). On multivariate analysis, the presence of metastatic disease was the only prognostic factor (P < 0.01). CONCLUSION: The VDC/ICE protocol was feasible, and considering the high tumor burden in our population, resulted in comparable results to those reported by cooperative groups in high-income countries. Further adaptation to maximize efficacy and minimize toxicity will be required.

Análise da mobilização e coleta de células-tronco hematopoiéticasdo sangue periférico (CTHSP) para transplante autólogo no HospitalUniversitário de Santa Maria, RS / Analysis of the mobilization and collection of peripheral blood stem cells (PBSC) for autologous transplantation in the University Hospital of Santa Maria, RS

Objetivo: Avaliar os resultados da mobilização e coleta de células-tronco hematopoiéticas do sangue periférico (CTHSP) e identificar as características dos pacientes atendidos na instituição. Métodos: Foram revisados retrospectivamente os prontuários de 176 pacientes com diagnósticos de Mieloma Múltiplo (MM), Linfoma de Hodgkin (LH),Linfoma não Hodgkin (LNH), Leucemia Mieloide Aguda (LMA) e outras neoplasias no período dedezembro de 1996 e 2011. Resultados: A mediana de idade dos pacientes foi de 42 anos, sendo 76,6% adultos e 23,3% pediátricos. Os diagnósticos mais frequentes foram MM (30,7%), LH (26,7%) e LNH (19,9%). Os métodos e regimes de mobilização incluíram o uso de G-CSF associado ou não a diferentes quimioterápicos: ciclofosfamida; ifosfamida, carboplatina e etoposide (ICE) e quimioterápicos relacionados ao tratamento do paciente. Foram realizadas 203 mobilizações e 474 coletas com média de 2,33 coletas porpaciente. O regime de mobilização com melhor rendimento de células CD34+ foi G-CSF associado a quimioterápicos relacionados ao tratamento do paciente. A mediana de rendimento de células nucleadas totais obtida no estudo foi 7,01x108/kg e de célulasCD34+ foi 2,63x106/kg. Dezoito pacientes (10,2%) não atingiram o número desejado após mobilização e coleta, sendo que a maioria dos pacientes (89,8%) atingiu a contagemde células CD34+ esperada. Conclusão: Constatou-se que fatores como o diagnóstico, o tratamento prévio de quimioterapia e radioterapia, o regime de mobilização, a contagem de leucócitos no início da coleta, os dias de administração de G-CSF e os diaspós-quimioterapia de mobilização necessitam ser considerados para a coleta de CTHSP, sendo que antes dos procedimentos cada paciente deve ser avaliado em relação à sua doença e fase de evolução.

Transplante autólogo de células-tronco do sangue periférico no Hospital Universitário de Santa Maria / Autologous peripheral blood stem cells transplantation at the University Hospital of Santa Maria

Introdução: O principal objetivo deste estudo foi identificar as características clínicas dos pacientes transplantados na instituição e avaliar os resultados obtidos com a infusão autóloga de células-tronco hematopoiéticas do sangue periférico (CTHSP), a mortalidade relacionada ao transplante (MRT) e a sobrevida global (SG). Métodos: Através da revisão e avaliação retrospectiva dos prontuários dos 120 pacientes submetidos a transplante autólogo no período de dezembro de 1996 a dezembro de 2011. Resultados: Cento e vinte pacientes receberam quimioterapia mieloablativa e resgate com infusões de CTHSP, sendo 78,3% adultos, com mediana de idade de 47 anos e predomínio do sexo masculino. Os diagnósticos foram 32,5% para Mieloma Múltiplo (MM), 35,8% para Linfoma de Hodgkin (LH), 16,7% para Linfoma não Hodgkin (LNH) 4,2% para Leucemia Mieloide Aguda (LMA) e 10,8% para outras neoplasias como Tumor de Wilms, Câncer de Mama Neuroblastoma, Sarcoma de Ewing, Tumor de Testículo, Meduloblastoma, Macroglobulinemia, Amiloidose e Tumor de SNC. A mediana do número de células nucleadas totais infundidas foi de 6,46x108/kg e a de células CD34+ foi de 3,17x106/kg. A mediana de tempo para recuperação de neutrófilos foi de 10 dias e para plaquetas, de 12 dias. Foi encontrada uma correlação entre a quantidade de células CD34+ infundidas e a recuperação de neutrófilos e plaquetas. Para o grupo em geral, a MRT encontrada foi de 5%, e a probabilidade de SG em cinco anos de 55,1%. Conclusão: Os resultados obtidos com os transplantes autólogos em nossa instituição são semelhantes aos descritos na literatura internacional (AU)

Introduction: The aim of this study was to identify the clinical characteristics of patients transplanted in the institution and evaluate the results obtained with the autologous infusion of hematopoietic stem cells from peripheral blood (PBSC), transplant-related mortality (TRM) and overall survival (OS). Methods: A review and retrospective assessment of the charts of 120 patients who underwent autologous transplantation from December 1996 to December 2011. Results: One hundred and twenty patients received myeloablative chemotherapy and rescue with infusions PBSC, of whom 78.3% were adults, with a median age of 47 years and male predominance. The diagnoses were 32.5 % for Multiple Myeloma (MM), 35.8% for Hodgkin lymphoma (HL), 16.7 % for non-Hodgkin lymphoma (NHL), 4.2 % for Acute Myeloid Leukemia (AML ), and 10.8% for other cancers such as Wilms Tumor, breast cancer, neuroblastoma, Ewing's sarcoma, Testicular Tumor, medulloblastoma , macroglobulinemia , amyloidosis and CNS tumor. The median number of total nucleated cells infused was 6.46 x108/kg and of CD34+ cells was 3.17 x106/kg. The median time for neutrophil recovery was 10 days and for platelets 12 days. A correlation was found between number of CD34+ cells infused and recovery of neutrophils and platelets. For the overall group, the MRT was found to be 5% and the probability of OS at five years was 55.1 %. Conclusion: The results obtained with autologous transplantation at our institution are similar to those described in the international literature (AU)

[Dipodascus capitatus (Geotrichum capitatum): fatal systemic infection on patient with acute myeloid leukemia]. / Dipodascus capitatus (Geotrichum capitatum): infecção sistêmica fatal em paciente com leucemia mielocítica aguda.

The infections caused by Dipodascus capitatus are rare, and the treatment is difficult. We reported a case of a patient with acute myeloid leukemia. The fungus was first isolated from hemocultures, and the phenotypic identification was based on mycological methods. The genotyping was carried out by sequencing the region D1/D2 from 26 rDNA. The susceptibility tests were assayed by Etest® and by the microdilution technique. None of the antifungal treatments employed were effective. The patient died on day 17 after the mycological diagnosis. The authors discussed the emergence of such infections as well as the difficulty regarding the diagnosis and treatment.

Dipodascus capitatus (Geotrichum capitatum): infecção sistêmica fatal em paciente com leucemia mielocítica aguda / Dipodascus capitatus (Geotrichum capitatum): fatal systemic infection on patient with acute myeloid leukemia

As infecções causadas por Dipodascus capitatus são raras e de difícil tratamento. Aqui se relata um caso em paciente com leucemia mielocítica aguda. O isolamento fúngico ocorreu a partir de hemocultura e a identificação fenotípica baseou-se em métodos micológicos clássicos; a identificação genotípica foi realizada através do sequenciamento da região D1/D2 do 26 rDNA. Os testes de suscetibilidade foram realizados através do Etest® e microdiluição em caldo. A antifungicoterapia foi ineficaz, registrando-se óbito da paciente no 17° dia após o diagnóstico. Os autores comparam o caso com relatos similares e discutem a emergência destas infecções bem como suas dificuldades diagnósticas e terapêuticas.

The infections caused by Dipodascus capitatus are rare, and the treatment is difficult. We reported a case of a patient with acute myeloid leukemia. The fungus was first isolated from hemocultures, and the phenotypic identification was based on mycological methods. The genotyping was carried out by sequencing the region D1/D2 from 26 rDNA. The susceptibility tests were assayed by Etest® and by the microdilution technique. None of the antifungal treatments employed were effective. The patient died on day 17 after the mycological diagnosis. The authors discussed the emergence of such infections as well as the difficulty regarding the diagnosis and treatment.

Benefits of the intermittent use of 6-mercaptopurine and methotrexate in maintenance treatment for low-risk acute lymphoblastic leukemia in children: randomized trial from the Brazilian Childhood Cooperative Group--protocol ALL-99.

PURPOSE To describe event-free survival (EFS) and toxicities in children with low-risk acute lymphoblastic leukemia (ALL) assigned to receive either continuous 6-mercaptopurine (6-MP) and weekly methotrexate (MTX) or intermittent 6-MP with intermediate-dose MTX, as maintenance treatment. PATIENTS AND METHODS Between October 1, 2000, and December 31, 2007, 635 patients with low-risk ALL were enrolled onto Brazilian Childhood Cooperative Group for ALL Treatment (GBTLI) ALL-99 protocol. Eligible children (n = 544) were randomly allocated to receive either continuous 6-MP/MTX (group 1, n = 272) or intermittent 6-MP (100 mg/m(2)/d for 10 days, with 11 days resting) and MTX (200 mg/m(2) every 3 weeks; group 2, n = 272). RESULTS The 5-year overall survival (OS) and EFS were 92.5% +/- 1.5% SE and 83.6% +/- 2.1% SE, respectively. According to maintenance regimen, the OS was 91.4% +/- 2.2% SE (group 1) and 93.6% +/- 2.1% SE (group 2; P = .28) and EFS 80.9% +/- 3.2% SE (group 1) and 86.5% +/- 2.8% SE (group 2; P = .089). Remarkably, the intermittent regimen led to significantly higher EFS among boys (85.7% v 74.9% SE; P = .027), while no difference was seen for girls (87.0% v 88.8% SE; P = .78). Toxic episodes were recorded in 226 and 237 children, respectively. Grade 3 to 4 toxic events for groups 1 and 2 were, respectively, 273 and 166 for hepatic dysfunction (P = .002), and 772 and 636 for hematologic episodes (P = .005). Deaths on maintenance were: seven (group 1) and one (group 2). CONCLUSION The intermittent use of 6-MP and MTX in maintenance is a less toxic regimen, with a trend toward better long-term EFS. Boys treated with the intermittent schedule had significantly better EFS.

External auditory canal and middle ear relapse of acute promyelocytic leukemia treated with arsenic trioxide: case report and review of the literature.

Extramedullary involvement occurs infrequently in acute promyelocytic leukemia and is said to be more common after treatment with all-trans retinoic acid. We describe a 9-year-old girl who had an isolated external auditory canal and middle ear relapse after treatment with all-trans retinoic acid and chemotherapy. A patient with cytogenetically and molecularly confirmed acute promyelocytic leukemia developed isolated extramedullary relapse in the auditory canal and middle ear 4 years and 9 months after initial diagnosis, while in hematologic and molecular remission, successfully treated with arsenic trioxide alone.

Aspectos epidemiológicos, biotipologia e evolução do tratamento da leucemia linfocítica aguda na infância e adolescência no Rio Grande do Sul / Epidemiological aspects, biotipology and evolution of the treatment of acute lymphocytic leukemia in childhood and adolescence in Rio Grande do Sul

A Leucemia Linfocítica Aguda na infância e adolescência é uma neoplasia de precursores linfóides de natureza heterogênea. Foi a primeira neoplasia disseminada a tornar-se curável pela quimioterapia. No Brasil os estudos cooperativos para o seu tratamento foram iniciados em 1980 com a criação do primeiro protocolo para o tratamento desta leucemia, denominado Grupo Brasileiro para o Tratamento da Leucemia na Infância. Na seqüência destes estudos foram observadas Sobrevidas Livre de Eventos de 50%, 58% e 70% nos protocolos 80, 82 e 85 respectivamente. Durante as décadas de 1980 e 1990, com a divulgação dos excelentes resultados alcançados com os regimes dos protocolos do grupo Berlin Frankfurt Münster, uma série de instituições em nosso País passou a adotá-los. Apesar de termos conhecimento dos dados referentes a evolução dos pacientes protocolados no GBTLI e dos demais estudos divulgados pelas instituições de origem, não tínhamos, porém, uma avaliação epidemiológica nem o conhecimento dos índices de sobrevivência alcançados no estado do Rio Grande do Sul. Neste trabalho foram avaliados 1472 pacientes com LLA, 833 (56,59%) do sexo masculino, 639 (43,41%) do sexo feminino, com idades entre zero e 20 anos, média de 7,40 anos (desvio padrão 5,14) e mediana de 5,70 anos (amplitude 0,06 a 20,76), no período de 1980 a 2008 provenientes deste Estado. Os dados foram colhidos individualmente dos prontuários dos pacientes admitidos nas principais instituições hospitalares que mantém assistência a pacientes pediátricos com neoplasias hematológicas. No presente estudo 487 pacientes (39,40%) foram registrados oficialmente nos protocolos do GBTLI; 678 (54,85%) receberam tratamento baseados nos regimes do grupo BFM e 71 (5,75%) por outros regimes incluindo os tratados segundo o protocolo UKALL. Os casos não foram, no entanto, protocolados e, na grande maioria não foram observados a totalidade dos itens exigidos para os pacientes registrados oficialmente...

BACKGROUND: The cryopreservation of parathyroid tissue is employed in the surgical treatment of secondary hyperparathyroidism in patients with chronic kidney disease. During the period between surgical resection and cryopreservation of tissue, which requires a specialized laboratory, the tissue is stored in a cell culture solution, at 4 °C (solution for transport from the operating room to the laboratory). There is no data showing for how long the human parathyroid tissue can remain viable in this solution, before being cryopreserved. The present study evaluates the time that the tissue of human hyperplastic parathyroid gland could remain in solution for transportation, without showing ultrastructural damages. METHODS: This prospective study included 11 patients, who underwent total parathyroidectomy with heterotopic autotransplantation and cryopreservation of parathyroid tissue fragments. Part of the tissue intended for pathological examination was selected for storage at solution for transportation. Five periods were defined, related to the storage time of parathyroid fragments at solution for transportation. At time 1, the material was fixed at the time of surgical resection, without contact with the solution for transport, this time was used as control. At time 2, the fragments of tissue remained stored at the solution for transportation for 2 hours, at time 3, this period was 6 hours, and Times 4 and 5, corresponded to the parathyroid fragments stored in the transport solution for 12 and 24 hours, respectively. At the end of each period the fragments were removed from the transport solution and fixed with 2% glutaraldehyde, followed by preparation of material for ultrathin sections. The analysis by electron microscopy was used to evaluate cell adhesion and integrity of plasma membranes, nuclei and mitochondria, and the presence of edema and cell vacuoles. RESULTS: Of the 11 cases studied, 10 showed ultrastructural findings consistent with the normal...

Aspectos epidemiológicos, biotipologia e evolução do tratamento da leucemia linfocítica aguda na infância e adolescência no Rio Grande do Sul / Epidemiologic aspects, biotypology and evolution of acute lymphocytic leukemia in childhood and adolescence in Rio Grande do Sul

A leucemia linfocítica aguda na infância e adolescência é umaneoplasia de precursores linfoides de natureza heterogênea. NoBrasil, os estudos cooperativos foram iniciados em 1980. Nasequência destes estudos foram observadas sobrevidas livre deeventos de 50%, 58% e 70% nos protocolos 80, 82 e 85 respectivamente.Neste trabalho foram avaliados 1.472 pacientes comLLA, 56,59% masculinos e 43,41% femininos, com idades entre 0e 20 anos, média de 7,40 anos, no período de 1980 a 2008 provenientesdo RS. Os dados foram colhidos dos prontuários dos pacientesdas principais instituições hospitalares que assistemneoplasias hematológicas pediátricas. Neste estudo, 487 pacientes(39,40%) foram registrados nos protocolos do GBTLI; 678(54,85%) receberam tratamento baseados nos regimes do grupoBFM e 71 (5,75%) por outros regimes. A sobrevida livre de eventosdos pacientes protocolados foi significativamente superiorcomparada aos não protocolados, 62,41% ± 2,43% e 53,86% ±2,04% respectivamente, em cinco anos. Pacientes com idade de 15a 19 anos tiveram um índice de SLE de 37,98% ± 4,72% em cincoanos, inferior quando comparado aos de 0 a 4 anos e 5 a 9 anosrespectivamente: 62,78% ± 2,28% e 62,43% ± 2,84%. Foi observada,na população estudada, uma SG de 63,73% ± 1,49% e SLEde 57,27% ± 1,57%. A incidência da LLA com progenitores Bseguiu o padrão observado em países desenvolvidos com um picode frequência absoluta entre as idades de 2 a 4 anos. Houve diferençasignificativa entre a população da região urbana ou rural:SLE em cinco anos de 61,76% ± 1,76% e 49,81% ± 4,28% respectivamente.A SLE e a SG em lactentes e portadores de síndrome deDown foi inferior aos resultados obtidos em instituições dos paísesdesenvolvidos.

Acute lymphocytic leukemia (ALL) in childhood andadolescence is a neoplastic disease of varied lymphoid precursors.In Brazil, cooperative studies on the treatment of ALL started in1980. According to these studies, the event free survival (EFS)was 50%, 58% and 70% in the protocols of 1980, 1982 and 1985,respectively. In this work, 1472 ALL patients were evaluated;56.59% were male and 43.41% were female with ages that rangedbetween 0 and 20 years old (mean age of 7.4 years old). Data wascollected from the records of patients with hematologic neoplasiasin medical institutions that offered treatment for pediatricneoplasias in the period from 1980 to 2008. In this study, 487patients (39.40%) were registered in the GBTLI protocol; 678(54.85%) received treatment based on the BFM group regimenand 71 (5.75%) were treated according to other regimens. TheEFS at five years of the patients on the GBTLI protocol wassignificantly higher than those who were not on this protocol(62.41% ± 2.43% and 53.86% ± 2.04%, respectively). In respectto the age range, the patients who were between 15 and 20 yearsold had an EFS of 37.98% ± 4.72% at five years, which is lowerthan the rate for 0 to 4-year-old and 5 to 9-year-old patients(62.78% ± 2.28% and 62.543± 2.84%, respectively). An overallsurvival of 63.73% ± 1.49% and EFS of 57.27% ± 1.57% wereobserved in the studied population. Epidemiologically, theincidence of ALL in B progenitors followed the pattern observedin developed countries with an absolute frequency peak in the agerange of 2 to 4 year olds. The outcome was better for patientsfrom urban areas compared to those from rural areas. EFS andoverall survival in infant and Down syndrome patients were worsethan the results obtained in developed countries.

Estimated number of cases, regional distribution and survival of patients diagnosed with acute myeloid leukemia between 1996 and 2000 in Rio Grande do Sul, Brazil.

Rio Grande do Sul (RS), in South Brazil, with about 10 million inhabitants, is known for its agricultural activities and consequent increased human exposure to toxic agents. Patients with de novo acute myeloid leukemia (AML) were included based on information retrieved from all referral hospitals in RS between 1996 and 2000. A total of 532 patients were registered. Median age at diagnosis was 42 years. The estimated annual incidence was 1.11 cases/100,000 inhabitants/year. There was an estimated incidence of 0.5-1 case per 100,000 inhabitants up to the age of 45 years, and of 3.5 cases per 100,000 inhabitants aged 70 years and older, with no geographical clusters. The mean 5-year survival rate was 17% for all cases. There was an increased number of M3 cases, as already described for individuals of Latin-American and the mortality rate was similar to that described in the literature.

The association of ICAM-1 Exon 6 (E469K) but not of ICAM-1 Exon 4 (G241R) and PECAM-1 Exon 3 (L125V) polymorphisms with the development of differentiation syndrome in acute promyelocytic leukemia.

The use of all trans-retinoic acid (ATRA) is the basis of treatment of acute promyelocytic leukemia (APL) and represents the paradigm of differentiation therapy. In general, ATRA is well-tolerated but may be associated with a potentially lethal side-effect, referred to as retinoic acid or differentiation syndrome (DS). The cellular and molecular mechanisms of DS are poorly understood and involve changes in the adhesive qualities and cytokine secretion of leukemic cells during ATRA-induced differentiation. As leukocyte extravasation is a key event in DS pathogenesis, we analyzed the association between the polymorphisms at Exon 4 (G241R) and Exon 6 (E469K) of ICAM-1 and Exon 3 (L125V) of PECAM-1 genes with DS development in APL patients treated with ATRA and anthracyclines. DS was diagnosed in 23/127 (18.1%) APL patients at an average of 11.5 days after the start of ATRA. All patients presented respiratory distress associated with increased ground-glass opacity in chest radiographies. Other accompanying symptoms were: fever not attributable to infection (65.2%), generalized edema (37.5%), weight gain (37.5%), and impairment of renal function (8.6%). We detected an association between development of DS and the AA genotype at Codon 469 of ICAM-1 (odds ratio of 3.5; 95% confidence interval: 1.2-10.2). Conversely, no significant association was detected between G241R or L125V polymorphisms at Exon 4 of ICAM-1 and Exon 3 of PECAM-1, respectively. Our results suggest that susceptibility to DS in APL patients may be influenced by genetic variation in adhesion molecule loci.

High frequency of t(12;21)(p13;q22) in children with acute lymphoblastic leukemia and known clinical outcome in southern Brazil.

The presence of the t(12;21)(p13;q22) distinguishes a subset of children with acute lymphoblastic leukemia (ALL) that present a favorable prognosis. This is a cryptic translocation difficult to detect through conventional cytogenetics. In this study, bone marrow samples from 30 children with ALL from southern Brazil were evaluated by fluorescence in situ hybridization (FISH) for the t(12;21), using locus specific probes to detect the TEL/AML1 rearrangement. The selection criteria included: age (0-12 years old); FAB classification (L1 or L2), absence of specific clonal chromosomal aberrations; and adequate cellular integrity to perform FISH analysis. A frequency of 40% of the t(12;21) was observed, in addition to extra copies of the AML1 gene in 7.5% of patients. These findings were analyzed in relation to the patient's clinical parameters and compared with other pediatric populations.

Abordagem na assistência à criança com câncer do Serviço de Hemato/Oncologia do Hospital Universitário de Santa Maria: percepção dos profissionais e voluntários / Approaching in the assistance to the child with cancer of the Serviço de Hemato/Oncologia of the Hospital Universitário de Santa Maria: perception of the professionals and volunteers

Sabe-se que a abordagem na assistência à saúde da criança é centrada na patologia, no sujeito ou no sujeito e sua família. Com o objetivo de identificar qual a percepção dos profissionais e voluntários sobre a abordagem de assistência à criança hospitalizada no Serviço de Hemato/Oncologia do HUSM, elaborou-se um questionário com 33 questões fechadas e 16 questões abertas, que foi enviado aos funcionários e voluntários que atuam no referido serviço, tendo sido garantido o anonimato. Foram devolvidos 54 questionários dos 120 enviados. Para análise das informações utilizou-se o modelo Processo-Pessoa-Contexto combinado com Cronossistema, da Teoria dos Sistemas Ecológicos de Bronfenbrenner. Com relação ao contexto, as respostas foram categorizadas de acordo com os parâmetros do contexto (microsistema, mesosistema, macrosistema) e considerando o modelo de Cronossistema propomos a análise nos tempos: no diagnóstico, durante o tratamento e após alta do tratamento. Verificou-se que o modelo de assistência à criança empregado tem uma tendência a ser centrado na criança e família, pois 47 por cento dos consultados informam que sua atividade profissional está direcionada à criança e sua família; 36 por cento referem-se que seu foco de atenção é a criança e apenas 6 por cento referem-se que sua assistência é centrada na patologia.