ALBERT

Description: This is a retrospective analysis of 1084 patients who received an allogeneic HSCT in 10 Brazilian Centers, from February 1983 to March 2003, aiming to validate the EBMT risk score. Data from transplanted patients and donors regarding patients age, disease stage at transplantation, HLA full-match sibling donor or full-match unrelated donors, donor-recipient gender match and the interval from diagnosis to transplantation, were used as variables to calculate the EBMT risk score. This score was analyzed using Cox Proportional Hazards Model. The OS, DFS, TRM and relapse were analyzed using Kaplan-Meier and cumulative incidence, whenever appropriate. In all there were 647 (60%) males and 437 (40%) females, the median age was 32 years old (range 1 – 59); 898 (83%) were in chronic phase, 146 (13%) were in accelerated phase and 40 (4%) were in blast crisis; 151 (14%) were younger than 20 years old, 620 (57%) were between 20 and 40 and 313 (29%) were older than 40; 1025 (95%) received HLA full-match sibling transplant and only 59 (5%) received an unrelated transplant. Female donor to male recipient occurred in 283 (26%) transplants. The interval from diagnosis to transplantation was less than 12 months in 223 (21%) cases and greater in 861 (79%). The OS, DFS, TRM and, relapse were 49%, 50%, 45%, 25%, respectively. The risk score 0–1 occurred in 179 (17%), score 2 in 397 (37%), score 3 in 345 (32%), score 4 in 135 (12%), and score 5–6 in 28 (2%). The risk scores 0–1 and 2 did not show any difference in terms of OS (58% and 55%, respectively) but they were significantly better than scores 3 or more (score 3 – 44%, 4 – 36 % and, 5-6 - 27%, respectively) (P

Description: Uncertainty still exists with the effects of allogeneic PBSCT on the clinical outcomes of patients with hematological malignancies. Our aim was analyzed retrospectively the clinical outcomes of 483 patients who underwent an allo PBSCT in 9 Brazilians centers from May 1994 to February 2004. The analyzes included patients with hematological malignancies who underwent PBSCT from HLA identical siblings donors treated with G-CSF at a dose of 10mgr/kg/day, 5 days. Median age was 34ys old (2–57), advanced disease was present in 58%, conditioning without irradiation was 85%; GVHD prophylaxis with MTX/CsA was 91%; CD34+ median was 4.7X106/kg (0.51–71.6); the median follow-up for surviving patients was 797 days (8–3420); median day for neutrophils and platelets engraftment was 15 and 14, respectively; cumulative incidence (CI) for ³ 2 aGVHD was 38%; extensive cGVHD 63%; CI for transplant relate mortality (TRM) 59%; CI for relapse 37%; the estimates of OS and DFS at 9 ys are 33% and 42, respectively. The following factors were significantly associated with better outcomes for engraftment, aGVHD, cGVHD, OS, DFS, relapse, and TRM in univariate analyzes, respectively: neutrophils and platelets engraftment: CD34+ cell dose 〉 2.8X106/kg, GVHD prophylaxis other than MTX/CsA, and sex match other than female donor for male recipient; aGVHD: age 4.7X106/kg; cGVHD: age 〈 25ys, sex match other than female donor for male recipient, advanced disease and CD3+ dose 〈 170X106/kg; OS: early disease; DFS: early disease, CD34+ dose 〉 4.7X106/kg; relapse: age〉 25ys, CD34+ cell dose 〉 2.8X106/kg, and early disease; TRM: early disease. All the results remained significant in multivariate analyzes, but CD34+ dose and platelet engraftment; age and CD34+ dose in aGVHD; age and CD3+ in cGVHD, and age in relapse. In our experience, sex match, CD34+ dose, GVHD prophylaxis may influence the engrafment, and sex match and disease phase the cGVHD. Furthermore, advanced disease had a negative impact on OS and TRM;. CD34+ higher dose and early disease were associated with better DFS and lower relapse.

Description: The safety and efficacy of administering ex vivo expanded peripheral blood progenitor cells (PBPC) to patients with breast cancer who undergo high-dose chemotherapy and PBPC transplantation was investigated. Unselected PBPC were cultured in gas-permeable bags containing 1-L serum-free media, granulocyte colony-stimulating factor, stem cell factor, and pegylated megakaryocyte growth and development factor for 9 days. Cell dose cohorts were assigned to have between 2 and 24 × 109 PBPC cultured at 1, 2, or 3 × 106 cells/mL. Twenty-four patients received high-dose chemotherapy followed by infusion of the cultured PBPC and at least 5 × 106 CD34+ uncultured cryopreserved PBPC per kilogram. No toxicities resulted from infusions of the ex vivo expanded PBPC. The study patients had shorter times to neutrophil (P = .0001) and platelet (P = .01) recovery and fewer red cell transfusions (P = .02) than 48 historical controls who received the same conditioning regimen and posttransplantation care and at least 5 × 106CD34+ PBPC per kilogram. Improvements in all these endpoints were significantly correlated with the expanded cell dose. Nine of 24 (38%) patients recovered neutrophil counts above 500/μL by day 5 or 6 after transplantation, whereas none of the controls had neutrophil recovery before the eighth day. Seven (29%) patients had neutropenia for 3 or fewer days, and 9 (38%) patients did not experience neutropenic fevers or require broad-spectrum antibiotics. Therefore, ex vivo expanded PBPC are capable of ameliorating posttransplantation neutropenia, thrombocytopenia, and anemia in patients receiving high-dose chemotherapy.

Description: The safety and efficacy of administering ex vivo expanded peripheral blood progenitor cells (PBPC) to patients with breast cancer who undergo high-dose chemotherapy and PBPC transplantation was investigated. Unselected PBPC were cultured in gas-permeable bags containing 1-L serum-free media, granulocyte colony-stimulating factor, stem cell factor, and pegylated megakaryocyte growth and development factor for 9 days. Cell dose cohorts were assigned to have between 2 and 24 × 109 PBPC cultured at 1, 2, or 3 × 106 cells/mL. Twenty-four patients received high-dose chemotherapy followed by infusion of the cultured PBPC and at least 5 × 106 CD34+ uncultured cryopreserved PBPC per kilogram. No toxicities resulted from infusions of the ex vivo expanded PBPC. The study patients had shorter times to neutrophil (P = .0001) and platelet (P = .01) recovery and fewer red cell transfusions (P = .02) than 48 historical controls who received the same conditioning regimen and posttransplantation care and at least 5 × 106CD34+ PBPC per kilogram. Improvements in all these endpoints were significantly correlated with the expanded cell dose. Nine of 24 (38%) patients recovered neutrophil counts above 500/μL by day 5 or 6 after transplantation, whereas none of the controls had neutrophil recovery before the eighth day. Seven (29%) patients had neutropenia for 3 or fewer days, and 9 (38%) patients did not experience neutropenic fevers or require broad-spectrum antibiotics. Therefore, ex vivo expanded PBPC are capable of ameliorating posttransplantation neutropenia, thrombocytopenia, and anemia in patients receiving high-dose chemotherapy.

Description: Heme, a ubiquitous iron-containing compound, is present in large amounts in many cells and is inherently dangerous, particularly when it escapes from intracellular sites. The release of heme from damaged cells and tissues is supposed to be higher in diseases such as malaria and hemolytic anemia or in trauma and hemorrhage. We investigated here the role of free ferriprotoporphyrin IX (hemin) as a proinflammatory molecule, with particular attention to its ability to activate neutrophil responses. Injecting hemin into the rat pleural cavity resulted in a dose-dependent migration of neutrophils, indicating that hemin is able to promote the recruitment of these cells in vivo. In vitro, hemin induced human neutrophil chemotaxis and cytoskeleton reorganization, as revealed by the increase of neutrophil actin polymerization. Exposure of human neutrophils to 3 μM hemin activated the expression of the chemokine interleukin-8, as demonstrated by quantitative reverse-transcription polymerase chain reaction, indicating a putative molecular mechanism by which hemin induces chemotaxis in vivo. Brief incubation of human neutrophils with micromolar concentrations of hemin (1-20 μM) triggered the oxidative burst, and the production of reactive oxygen species was directly proportional to the concentration of hemin added to the cells. Finally, we observed that human neutrophil protein kinase C was activated by hemin in vitro, with a K1/2 of 5 μM. Taken together, these results suggest a role for hemin as a proinflammatory agent able to induce polymorphonuclear neutrophil activation in situations of clinical relevance, such as hemolysis or hemoglobinemia.

Description: Purpose: Imatinib Mesilate is currently considered the most advanced therapy for CML due to its innovative molecular targeting mechanism of action, resulting in a high level of efficacy with a favorable toxicity profile which results in health-related quality of life (HRQL) benefits. We therefore decided to evaluate 230 patients with chronic myeloid leukemia (CML) treated with Imatinib Mesilate (Gleevec) as second line therapy, along 12 months, after failing Interferon (IFN) based therapy, due to lack of efficacy or toxicity, in an a Quality of Life (QoL) Study in Brazil, here presented as an interim analysis. Patients and Methods: From July 2002 until December 2003, 230 Patients from 21 Brazilian Medical Institutions, were enrolled in a Quality of Life (QoL) Study, where patients completed FACT-BRM questionnaire at baseline and during treatment. FACT-BRM includes 4 subscales appropriate for any chronic illness or treatment [physical (PWB), social/family (SFWB), emotional (EWB), functional (FWB)] and 2 treatment-specific subscales [BRM-physical (BRMP). The primary endpoint was the composite Trial Outcomes Index (TOI=PWB + FWB +BRMP + BRMCE). The questionnaire was applied at baseline (visit 0),monthly thereafter during the first 6 months on imatinib therapy (visits 1 through 7) and finally after 12 months on treatment (visit 8). Study population was distributed as following: gender 55,9% male and 44,1% female, mean age 46 years (range 18 to 76), Karnofsky status 100% in 56.7% and 80–90% in 29.9%. Initial Imatinib Mesilate dose was 400mg for 81,3% and 600mg for 18,7%. All patients were evaluated at baseline and at visits 7 (six months) and 8 (12 months) 147 (63.9%) and 45 (19.5%) patients were evaluated, respectively. Results: An increase of 5 or more from baseline was defined as a clinically relevant improvement 6. Patients had clinically relevant mean estimated improvement in TOI along treatment. After 1 treatment month TOI had an estimated mean increase of 5.4 (p

Description: Transcription factors, including GATA-1 protein, play a key role in controlling the cellular proliferation and differentiation of hematopoietic stem cells. Inherited missense mutations in exon 4 of GATA-1 have been found in some families, leading to a variable degree of macrothrombocytopenia with or without abnormalities in the erythrocyte lineage. Acquired mutations in exon 2 of GATA-1 have been found in Down syndrome patients with transient myeloproliferative disorder (TMD) or megakaryoblastic leukemia (AMKL). These mutations prevent the synthesis of the full length GATA-1, but allow the synthesis of a smaller form of the protein (GATA-1s). Here, we report a novel inherited mutation, in exon 2 of the GATA-1 gene, which allows only the synthesis of GATA-1s in members of a family, who present anemia and neutropenia. A man, 19 years, was referred to our hospital due to anemia manifested at 4 years of age. Blood count showed: RBC: 3.0 x106/μl, HGB: 2.9g/dl, HCT: 8.3%, MCV: 94.4fl, MCH: 32.7pg, Retic: 0.6%, WBC: 2.6x103/μl, neutrophil: 1.0x103/μl, and PLT: 250.0x103/μl. The peripheral blood film demonstrated anisocytosis, macrocytosis, poikilocytosis, and neutrophils with pseudo Pelger-Hüet anomaly. The bone marrow aspirate and biopsy showed moderate hypocellularity with trilineage dysplasia. The presumptive diagnosis of hypocellular MDS was established. Karyoptype: 46, XY. DEB: negative. CFU-GM: 54 colonies/ml (normal range: 0.5 x104 to 1.0x104). Platelet aggregation test: 0.82 mg/ml (normal range: 0.7 to 1.2), platelet aggregation to 3μM adrenaline: absent (normal: present), and platelet aggregation to 3μM ADP: second wave absent (normal: present). The number of platelets expressing GPIX glycoprotein as well as those expressing GPIb, GPIIb, and GPIIIa were similar in patient and controls. No significant differences in the mean fluorescence indexes (MFI) for GPIX (1,214.0 vs 1,015.5), GPIb (154.0 vs 136.5), and GPIIb (100.0 vs 96.1) platelet contents were found in patient and controls. In contrast, the MFI for measurement of GPIIIa was slightly lower in patient than in controls (1,538.6 vs 1,928.1). Hypocellular MDS and similar laboratory findings were also identified in 6 additional males of the same family. Hemizygous and heterozygous 332G/C mutations in exon 2 of GATA-1 were found by sequencing PCR products in 7 males and in 4 phenotypically normal females of the family, comprising three generations. Only GATA-1s, probably resulting from alternative splicing of exon 2 was identified by RT- PCR and sequencing in affected males. In heterozygous females and normal members, both isoforms of the protein were found. These results indicate that the inherited 332G/C mutation in GATA-1 is associated with a clinical and hematological picture similar to that of MDS syndrome in contrast with the other mutations in exon 2 observed in Down syndrome, which are associated with AMKL or TMD.

Description: Multidrug resistance (MDR) to chemotherapeutic drugs is a major obstacle in the treatment of patients with cancer. One of the best characterized resistance mechanisms of resistance in tumors cells is the MDR mediated by P-glycoprotein (Pgp) and multidrug resistant related protein (MRP). MDR expression in non-Hodgkin lymphoma (NHL) range widely being dependent on factors as the different antibodies, detection methods used, and clinical status (untreated patients or after failure of chemotherapy). AIDS-related NHL is associated with high rate of relapse and short overall median survivals. Previous study demonstrated that MDR positive AIDS-related LNH patients had a lower complete remission compared with MDR negative patients. In this study we analysed expression of Pgp and MRP by immunohistochemistry (IHC) in 62 high grade B-cell types NHL in an attempt to determine if MDR expression is dependent on previous anti-HIV therapy, a group of drugs well-known substrates for MDR expression. IHC detection of Pgp and MRP was performed using monoclonal antibodies according avidin-biotin method. Expression of Pgp was observed in 30 of 62 (48.4%) cases of AIDS-related NHL. MRP expression was examined in 27 specimens being 7 cases considered positives (25.9%). Co-expression of Pgp and MRP was detected in 5 out of 27 (18.5%) samples, whose patients had not previous NHL diagnosis. Anti-HIV therapy was used in 31/62 patients prior NHL diagnosis. Sixteen out of 30 patients that exhibited positive Pgp cells had not received anti-HIV therapy before NHL. Our study shows that expression of Pgp did not correlate to previous anti-HIV therapy and can be also observed in newly diagnosed AIDS-related NHL. The biological linkages between NHL and MDR phenotype are not fully understood. However, our results showed an intrinsic resistance in these patients indicating that the MDR modulation during chemotherapy could be a new approach for treatment in AIDS-related NHL.

Description: As mouse models have become commonplace for studying hemostasis and thrombosis, we considered whether the mouse system had utility for assessing genetic alterations in platelet receptors. Platelets from 5 mouse strains (C57BL/6 [C57], FVB/N [FVB], BALB/c, C3H/He, and 129Sv) showed only minor differences in the expression of integrin αIIb, integrin β3, glycoprotein (GP) Ibα, or GPVI across strains. However, FVB platelets expressed approximately 50% the level of integrin α2 as platelets from other strains (P 〈 .0001). We bred FVB mice with C57 and assessed α2 expression in FVB/C57xFVB/C57 (F2) offspring. Linkage analysis demonstrated the gene responsible for α2 levels is tightly linked to the D13mit260 marker (log odds [lod] score 6.7) near the α2 gene. FVB platelets showed reduced aggregation and a longer lag phase to collagen. FVB and C57 platelets aggregated similarly to collagen-related peptide, but FVB platelets showed a reduction in rhodocytin-induced Syk and PLCγ2 tyrosine phosphorylation. Thus, FVB platelets express half the level of α2 as other mouse strains, a trait linked to the α2 gene and seemingly responsible for reduced platelet aggregation to collagen. These strain differences serve as a useful model for the 2-fold difference in human platelet α2β1 expression and demonstrate that α2β1 participates in signaling during platelet activation. (Blood. 2004;103:3396-3402)

Description: In 1997, a measles outbreak was identified in São Paulo. Between February and December, 20 185 cases were confirmed. From April to July 1997, a seroepidemiologic survey was conducted to identify the recipients of bone marrow (BM) transplants who were susceptible to measles and the occurrence of measles in this population. A total of 156 patients were screened by enzyme immunoassay (EIA). Patients with IgG titers more than 100 mIU/mL were considered immune. Measles reimmunization records were also reviewed. Thirty-two vaccinated patients underwent serologic evaluation. Six of 22 patients (27.3%) within 3 years after vaccination lost measles immunity, in contrast to 7 of 10 patients (70%) vaccinated longer than 3 years previously (P = .049). Among the 122 nonvaccinated patients, 41 (33.6%) were susceptible to measles: 4 of 47 patients (8.5%) within the first year after BM transplantation (BMT), and 37 of the 75 patients (49.3%) after the first year after BMT (P