Standardization of a multiplex assay to identify weak D types in a mixed-race Brazilian population.

RH allele variability is caused by several types of variants, resulting in altered RhD and RhCE phenotypes. Most of the weak D phenotypes in European-derived populations are weak D types 1, 2, or 3, which are not involved in alloimmunization episodes. However, the Brazilian population is racially diverse, and the accuracy of molecular and serologic tests developed in recent years has allowed for the identification of other RH variants, that are common in the Brazilian population, such as weak D type 38 or weak partial 11, the latter involved in alloimmunization cases. Furthermore, patients with these two weak D variants must be transfused with D- red blood cell units, as do patients with weak D type 4 or DAR, which are also common D variants in Brazil. Weak D type 38 and weak partial 11 can be serologically misclassified as weak D types 1, 2, or 3 in patients, based on European experience, or as D- in donors. Additionally, pregnant women may unnecessarily be identified as requiring Rh immune globulin. RhCE phenotypes are reliable indicators of RhD variants. For individuals with the Dce phenotype, the preferred approach is to specifically search for RHD*DAR. However, when encountering DCe or DcE phenotypes, we currently lack a developed method that assists us in rapidly identifying and determining the appropriate course of action for the patient or pregnant woman. Two multiplex assays were proposed: one for the identification of RHD*weak partial 11, RHD*weak D type 38, and RHD*weak D type 3 and another for RHD*weak D type 2 and RHD*weak D type 5. The multiplex assays were considered valid if the obtained results were equivalent to those obtained from sequencing. Expected results were obtained for all tested samples. The proposed multiplex allele-specific polymerase chain reaction assays can be used in the molecular investigation of women of childbearing age, patients, and blood donors presenting a weak D phenotype with DCe or DcE haplotypes in a mixed-race population, such as Brazil.

Comparative analysis of the pathological events involved in immune and non-immune TRALI models.

BACKGROUND AND OBJECTIVES: Transfusion-related acute lung injury (TRALI) is characterized by leukocyte transmigration and alveolar capillary leakage shortly after transfusion. TRALI pathogenesis has not been fully elucidated. In some cases, the infusion of alloantibodies (immune model), whereas in others the combination of neutrophil priming by proinflammatory molecules with the subsequent infusion of biological response modifiers (BRMs) in the hemocomponent (non-immune model) have been implicated. Our aim was to compare the pathological events involved in TRALI induced by antibodies or BRMs using murine models. MATERIALS AND METHODS: In the immune model, human HNA-2(+) neutrophils were incubated in vitro with a monoclonal antibody (anti-CD177, clone 7D8) directed against the HNA-2 antigen and injected i.v. in NOD/SCID mice. In the non-immune model, BALB/c mice were treated with low doses of lipopolysaccharide (LPS) followed by platelet-activating factor (PAF) infusion 2 h later. Forty minutes after PAF administration, or 6 h after neutrophil injection, lungs were isolated and histological analysis, determination of a variety of cytokines and chemokines including keratinocyte-derived chemokine (KC), MIP-2, the interleukins IL-1ß, IL-6, IL-8 as well as TNFα, cell influx and alveolar capillary leakage were performed. RESULTS: In both models, characteristic histological findings of TRALI and an increase in KC and MIP-2 levels were detected. In contrast to the immune model, in the non-immune model, there was a dramatic increase in IL-1ß and TNFα. However, capillary leakage was only detected if PAF was administrated. CONCLUSIONS: Regardless of the triggering event(s), KC, MIP-2 and integrins participate in TRALI pathogenesis, whereas PAF is essential for capillary leakage when two events are involved.

RHD alleles in Brazilian blood donors with weak D or D-negative phenotypes.

The RHD gene is highly polymorphic and the existence of a large number of alleles results in RhD variant phenotypes. RHD genotyping has been used to distinguish normal D antigen from D variants due to limitations of serologic methods. The purpose of this study was to determine the phenotypic frequency of RhD and RhCE antigens and to investigate the RHD alleles present in samples with the weak D or D- phenotypes from Brazilian blood donors. A total of 2007 donors were phenotyped for D, C, c, E and e antigens. Samples phenotyped as D- were genotyped by polymerase chain reaction-sequence specific primers, and exon 10 and intron 4 of the RHD gene were analysed. D- samples containing the RHD gene or samples considered weak D were further characterised using genotyping platform or nucleotide sequencing. Using serologic methods we found that 87.3% of the donors were D+, 11.9% D- and 0.8% weak D. The frequency of RHD gene in D- individuals was 9.2%. Five RHD alleles from phenotypically D- donors were characterised in six molecular backgrounds: RHDΨ, RHD-CE-D(s), RHD-CE-(2-9)-D, RHD/RHDΨ, RHDΨ/RHD-CE-D(s) and RHD-CE(2)-D. The most common weak D antigens types found were 1, 3, 4.0/4.1 and 4.2, whereas the most prevalent weak D type was 4.2 (or DAR). The RHD genotyping proved to be a necessary tool to characterise RHD alleles in donors phenotyped as D- or weak D to increase the transfusion safety in highly racial mixed population.

Molecular studies reveal that A134T, G156A and G1333A SNPs in the CD177 gene are associated with atypical expression of human neutrophil antigen-2.

BACKGROUND AND OBJECTIVES: The human neutrophil antigen-2 (HNA-2) is expressed on a subpopulation of neutrophils as most subjects present a negative plus a positive HNA-2 population of neutrophils. The number of neutrophils expressing HNA-2 is variable and may increase in pregnancy, infections, myeloproliferative disorders and after G-CSF. This study investigated the presence of polymorphisms in the gene encoding HNA-2 (CD177) in individuals presenting different patterns of antigen expression and determined the association of single nucleotide polymorphisms (SNPs) with the heterogeneous HNA-2 expression. MATERIALS AND METHODS: Flow cytometry was employed to analyse the HNA-2 expression on neutrophils from 135 healthy subjects using two monoclonal antibodies (TAG4, 7D8). Sequencing reactions were performed on subjects whose antigen expression was low (< or = 50%), high (> or = 80%) or atypical (a nonreactive population plus two distinct positive cell populations). RESULTS: Five SNPs were detected, two of them (A793C, G1084A) were related to a low expression of HNA-2 (P = 0.031 and P = 0.004). Atypical antigen expression was observed in 5.9% (8/135) of the individuals, three nonpregnant women and five men. In these cases, the cDNA sequences revealed three SNPs (A134T, G156A and G1333A) strongly related to this atypical HNA-2 expression (P = 0.004, P = 0.006 and P < 0.0001, respectively). CONCLUSIONS: Our data show that polymorphisms in the CD177 are associated with variations in the HNA-2 expression and may be the cause of atypical expressions.

Human neutrophil alloantigen-1a, -1b, -2, -3a and -4a frequencies in Brazilians.

Human neutrophil reactive antibodies may cause clinical disorders such as transfusion-related acute lung injury, febrile transfusion reactions, alloimmune neonatal neutropenia, immune neutropenia after stem cell transplantation, refractoriness to granulocyte transfusion, drug-induced neutropenia and autoimmune neutropenia. Using the granulocyte immunofluorescence test by flow cytometry, the phenotypic frequencies of the human neutrophil alloantigens (HNA)-1a, -1b, -2, -3a and -4a were determined in 100 healthy Brazilian persons. Neutrophils were separated from blood samples by sedimentation, centrifugated and incubated with HNA-specific alloantibody plus fluorescein isothiocyanate-labeled F(ab')(2) fragments of anti-human IgG. The results showed that the phenotype frequencies of HNA-1a, -1b, -2a, -3a and -4a were 65%, 83%, 97%, 95% and 94%, respectively. We detected that neutrophils from 17% of Brazilians typed positive only with anti-HNA-1a (HNA-1a/a), 35% only with anti-HNA-1b (HNA-1b/b) and 48% reacted with both antibodies (HNA-1a/b). The frequencies found for HNA-1a and -1b were quite similar to that reported among Africans and American-Africans, but different from those found in Japanese and Chinese. In addition, our data showed that the frequencies of HNA-2, -3a and -4a in Brazilians were comparable with those observed in Caucasians. The determination of HNAs frequencies among populations with distinct racial backgrounds is important not only for anthropological reasons, but also for neonatal typing in suspected cases of alloimmune neutropenia or when patients are severely neutropenic.

A very strict guideline reduces the number of erythrocyte transfusions in preterm infants.

BACKGROUND AND OBJECTIVES: Benefits of adopting restrictive guidelines for erythrocyte transfusions are still controversial. The objective of this study was to verify if a very strict guideline could reduce erythrocyte transfusions in preterm infants without adverse outcomes. MATERIALS AND METHODS: Two prospective cohorts of neonates with gestational age < 37 weeks and birth weight < 1500 g were studied. Neonates born in Period 1 were submitted to a strict guideline for erythrocyte transfusions. In Period 2, a new stricter protocol was introduced. Infants of both periods were compared regarding number of transfusions and clinical outcome. RESULTS: The median number of transfusions decreased from 2 (1 to 14) in Period 1 to 1 (1-9), P = 0.001, in Period 2. The linear regression multivariate analysis showed that the implementation of the stricter guideline was associated with a reduction in the number of transfusions received by patients by 0.55 (95% confidence interval: -0.08; -1.02) units/patients. Number of apnea episodes, weight at 28 days of life and days of hospital stay were similar in both periods. Intra-hospital death was lower in Period 2. CONCLUSION: A very strict guideline reduced the number of erythrocyte transfusions in preterm infants, without threatening their clinical course.

Anti-hepatitis C virus-positive blood donors: are women any different?

We sought to assess clinical, epidemiological, biochemical, serological and histological characteristics of anti-hepatitis C virus (HCV)-positive female blood donors and compare them with men. As women are frequently the minority among blood donors, studies evaluating this population usually reflect characteristics of male gender. This retrospective study included 380 blood donors with confirmed positive anti-HCV. The mean age was 36.9 +/- 11.3 years and 33.2% were women. Compared with men, female donors showed higher prevalence of prior transfusion of blood products (P = 0.031) and lower prevalence of intravenous drug use (P = 0.001) and alcohol abuse (P < 0.001). Women exhibited lower medians of alanine aminotransferase (P < 0.001) and gamma-glutamyltransferase (P < 0.001). They also showed higher platelet count (P < 0.001) and prothrombin activity (P = 0.049), and a lower frequency of antibody against core antigen of hepatitis B virus (anti-HBc) positivity (P = 0.032). A higher proportion of spontaneous viral clearance (P = 0.001) and a lower frequency of viraemia (P < 0.001) were observed among women. On liver biopsy, women had lower prevalence of fibrosis stage > or = 2. Multivariate analysis identified age (OR = 1.050, 95% CI: 1.019-1.081, P = 0.001) and anti-HBc positivity (OR = 2.184, 95% CI: 1.010-4.722, P = 0.047) as independent predictors of significant fibrosis. Female blood donors presented higher prevalence of spontaneous viral clearance as well as biochemical and histological evidence of less advanced liver disease. These findings could be because of intrinsic characteristics of female gender or secondary to associated factors such as younger age or anti-HBc positivity.

Analysis of polymorphism at site -174 G/C of interleukin-6 promoter region in multiple myeloma.

It is well established that interleukin-6 (IL-6) is an essential growth factor for multiple myeloma (MM) and patients with increased IL-6 levels have a poor prognosis. In healthy subjects, the presence of the C allele at a polymorphic site (-174 G/C) of the IL-6 gene is related to low IL-6 levels. In view of the potential association of this particular polymorphism with IL-6 concentration, and the relevance of IL-6 in MM pathogenesis, the objective of the present study was to investigate the prevalence of IL-6 (-174 G/C) promoter polymorphism and its association with development of MM in Brazilian individuals. We investigated the prevalence of these alleles in 52 patients and 60 healthy subjects (matched by age, sex, and race) of a Brazilian population. Thirty patients were male (42.4%), 24 (46.2%) were white and the median age at diagnosis was 58.5 years (range: 28 to 84 years). To determine the IL-6 (-174 G/C) polymorphism, molecular analysis was performed by polymerase chain reaction followed by endonuclease restriction digestion. The genotype distributions observed in the group of patients were 4% CC, 42% GC and 54% GG. The C allele frequency was 0.25. These results were similar to the control group, suggesting no impact of this polymorphism on the susceptibility to MM.

Plasmablastic multiple myeloma is associated with increased vascular endothelial growth factor immunoexpression.

The biologic basis of the negative prognosis of plasmablastic myeloma is not fully understood. To determine whether histologically aggressive multiple myeloma (MM) is associated with a more angiogenic marrow environment, bone marrow samples from 50 recently diagnosed MM patients were evaluated. Twelve percent (6/50) of patients presented plasmablastic MM, and this feature correlated with moderate/strong intensity of vascular endothelial growth factor staining of plasma cells (P = 0.036). Although plasmablastic MM was not associated with increasing of microvessel density, this new evidence of increased expression of vascular endothelial growth factor on plasmablasts suggests that the adverse prognosis conferred by plasmablastic disease may be due, at least in part, to secretion of this angiogenic cytokine, also suggesting that the subset of MM patients with plasmablastic features may derive particular benefit from antiangiogenic therapies.

Strategies for prevention of transfusion-associated Chagas' disease.

Available epidemiological data indicate that Chagas' disease, a zoonosis caused by the flagellate protozoan parasite T cruzi, is a very important medical and social problem in Latin America. More than 60% of T cruzi-infected individuals have migrated to urban areas, in both endemic and nonendemic countries. Thus, with the implementation and maintenance of regular vector control programs in some countries, allogeneic blood transfusions have been the main mechanism for the continuation of this endemy. The risk of infection after transfusion of a unit of T cruzi-infected blood product depends mainly on the amount of blood transfused, parasite concentration in the infected transfused blood unit, and the recipient's immunological status. Current strategies to prevent transfusion-associated Chagas' disease include the identification of T cruzi-infected blood donors by predonation questionnaire, serological tests for T cruzi antibodies, and the treatment of the blood collected with gentian violet. Because T cruzi infection is lifelong, and most infected persons are asymptomatic, the identification of high-risk blood donors by a predonation questionnaire is relevant in nonendemic countries but this strategy seems to be of limited usefulness for donor deferral in endemic areas. Because T cruzi antigens are shared by other parasites, the serological diagnosis of T cruzi infection is complex yielding both false-positive and false-negative results. Although sensitive, the tests currently available for the serodiagnosis of T cruzi infection lack specificity and a more specific, confirmatory test is still needed for the routine confirmation of T cruzi chronic infection. In areas of high endemicity or where serological screening is not available, the risk of T cruzi transmission by blood transfusion may be reduced by the addition of gentian violet to the collected blood. The use of gentian violet, alone or combined with ascorbic acid and light, effectively inactivate T cruzi present in donor blood; however, the long-term toxicity of this agent for blood recipients is still an open issue. In conclusion, the prevention of TA-CD is based on various strategies that are not mutually exclusive. Blood donor education, identification of putatively infectious blood donors by questionnaire or serological screening tests, and methods of parasite inactivation may significantly reduce the transmission of T cruzi by allogeneic blood transfusions.

Selectivity of human T lymphotropic virus type-1 (HTLV-1) and HTLV-2 infection among different populations in Brazil.

A seroprevalence study for human T lymphotropic virus type-1 (HTLV-1) and HTLV-2 was conducted in Sao Paulo, Brazil among 2,312 individuals that included following groups: 1,148 volunteer blood donors, 37 patients with tropical spastic paraparesis (TSP), 53 with lymphoproliferative disorders, 171 with a history of multiple blood transfusions, 268 human immunodeficiency virus type-1 (HIV-1) seropositive subjects, and 635 Amazonian Indians. Antibodies to HTLV-1/2 were screened by enzyme-linked immunosorbent assay (ELISA) and confirmed by Western blot and/or radioimmunoprecipitation. The differentiation of HTLV-1 and HTLV-2 was achieved using a synthetic recombinant peptide (rgp46) ELISA. We confirmed the presence of HTLV-1 infection in Brazil, both in blood donors (0.4%) and in patients exposed to blood transfusions (2.9%), as well as the occurrence of HTLV-1-associated TSP (11 patients, or 30% of all TSP cases) and adult T cell leukemia/lymphoma (two cases, or 3.5% of all hematologic malignancies). The HIV-1 infected individuals were shown to be coinfected (8.9%) with either HTLV-1 or HTLV-2. All HIV-1 and HTLV-2 coinfected individuals were intravenous drug abusers. In addition, we also demonstrated the presence of HTLV-2 (4.7%), and HTLV-1/2 (0.8%) in tribes of Amazonian Indians who lived in the eastern Amazon basin (southeastern State of Para). The selectivity of these retroviral infections in particular groups is emphasized, as well as the need for HTLV-1/2 screening of all blood donors in Brazil as a public health measure.

Immunosuppressive effects of allogeneic blood transfusions: implications for the patient with a malignancy.

A considerable amount of information has accumulated over the past decade indicating that the transfusion of allogeneic blood products may be associated with adverse effects to the recipient. These include the development of transfusion reactions, TA-GVHD, HLA-alloimmunization, and immunomodulatory effects. The latter might be beneficial for recipients of kidney allografts, in reducing the relapse rate in patients with Crohn's disease, and in ameliorating the rate of recurrence of spontaneous abortion in affected patients; however, the immunosuppressive effects associated with perioperative ABT might adversely affect overall prognosis in patients with a malignancy who undergo curative cancer surgery. In addition, ABT has been shown to be associated with an increased risk for postoperative bacterial infections. The ABT-induced immunomodulatory effects appear to be mediated immunologically by transfused allogeneic passenger leukocytes. The 3 log10 leukocyte reduction of cellular blood products, provided by the currently available commercial leukocyte filters, has been shown to minimize the occurrence of some of the ABT-associated deleterious effects; however, the actual clinical efficacy of leukodepletion has not yet been established, because the available data are largely from retrospective or uncontrolled clinical studies. Properly designed prospective clinical trials are essential to establish optimal conditions for the preparation of blood components destined for clinical use.

HTLV-I infection among relatives of patients with adult T-cell leukemia/lymphoma in Brazil: analysis of infection transmission.

We examined the presence of HTLV-I infection among 66 family members of 13 patients with well documented ATL to investigate the routes of HLTV-I transmission in a Southeast region of Brazil. HTLV-I infection was screened by an enzyme immunossay (ELISA) test and all repeatedly positive or indeterminate ELISA samples were further tested by a Western-Blot (WB) technique. Indeterminate and inconclusive WB samples were confirmed by a polymerase chain reaction (PCR). ELISA results showed that 40 (60.6%) individuals were not infected; 16 (24.2%) were positive; and 10 (15.2%) were undetermined. Among 16 ELISA positive subjects, 14 (87.5%) were confirmed to be positive by WB while 2 (12.5%) showed inconclusive results. Based on the laboratory data, questionnaire analysis, and family/epidemiological studies, we concluded that HTLV-I vertical transmission occurred in 6 of the 13 families. In 3 of these 6 families, the horizontal transmission also could be demonstrated. An isolated horizontal transmission was detected in one family, and in 6 families we did not find any infected family member. All HTLV-I-infected persons were clinically asymptomatic. The occurrence of an effective HTLV-I vertical transmission detected by the present study suggest that HTLV-I infection is endemic in the Southeast region of Brazil. Consistent with the modes of transmission, the HTLV-I antibody seroprevalence was greater in relatives of ATL patients than in the general blood donor Brazilian population (0.4%). In addition, the present data suggest that HTLV-I carries a high infectivity rate but a low virulence.

Importance of combined-modality therapy for primary bone lymphoma.

Primary bone lymphoma (PBL) is a rare entity and comprises about 5% of all extranodal non-Hodgkin's lymphomas (NHL) and 7% of all primary bone tumors. To date there is no consensus about the optimal treatment for PBL. We retrospectively reviewed all cases of PBL treated at Hospital São Paulo, Brazil, over a 10-year-period (January 1992-January 2002). Medical records of 7 patients with PBL were reviewed and information on age at diagnosis, sex, NHL clinical staging (CS), treatment and response to treatment were retrieved. Five patients (72%) received combined-modality therapy (CMT) and all of them are in complete remission (CR) with a median follow up of 19 months (ranging from 12 to 144 months). We conclude that PBL is a potentially curable malignancy and treatment should be undertaken in a multiprofessional approach, in order to provide the best support which probably has to include chemotherapy, radiotherapy and, for patients with IPI higher than 2, consolidation with stem-cell transplantation.

Level of fetal hemoglobin as an indicator of clinical complications in sickle cell anemia.

1. The fetal hemoglobin (HbF) level was used as an indicator of the development of severe clinical complications in 89 patients with sickle cell anemia (SCA). 2. HbF was determined by the alkali denaturation technique. The mean HbF level was 6.7 +/- 4.2% (range, 0.7 to 19.2%) of total hemoglobin. 3. Major organ failures were considered to be terminal events of morbidity and included 8 cerebrovascular accidents, 13 aseptic necroses of the femoral head and 17 leg ulcer episodes. 4. The characteristics of the test, including sensitivity, specificity and positive predictive value were analyzed for different levels of HbF. 5. The overall specificities were 76, 76, and 85% for HbF levels greater than or equal to 8, 10 and 12%, respectively. The sensitivity of the test was low. The positive predictive value reached 71% for children with HbF greater than or equal to 8%. The data suggest that HbF level may be a useful indicator of the possibility of a patient developing serious clinical complications.

Fetal hemoglobin in patients with chronic renal disease.

To investigate-whether hemoglobin (Hb) synthesis is affected by different treatment protocols used for end-stage renal disease, we analyzed the electrophoretic pattern of hemoglobin in 136 adult patients with chronic renal failure. Forty-seven patients were not in a dialysis program (ND), 29 individuals were on continuous ambulatory peritoneal dialysis (CAPD), 33 patients were on hemodialysis (HD), and 27 subjects had received a kidney transplant (KT). We found 3.6% hemoglobin C, 1.4% hemoglobin S and 3.6% B-thalassemia minor as reported in other studies of Brazilian patients. In addition, we found increased fetal hemoglobin (Hb F) levels in 7.4% of the patients which contrasts with the reported 0.01% prevalence rate of hereditary persistence of Hb F in Brazil. Seven out of ten patients with elevated Hb F belonged to either the CAPD or the KT group. We postulate that stress erythropoiesis is probably the mechanism responsible for the Hb F increase in these patients. However, properly designed clinical studies are still necessary to clarify these questions.

Measurement of red blood cell antibodies in autoimmune hemolytic anemia.

The enzyme-linked antiglobulin test (ELAT) was employed to measure the number of IgG molecules per red blood cell (IgG/RBC) in 11 patients with autoimmune hemolytic anemia (AIHA). All patients with AIHA had high levels of red cell-associated IgG (110-3, 650 IgG/RBC). The control group consisted of normal volunteers (N = 10) and patients with hereditary spherocytosis (N = 1), beta 0-thalassemia (N = 1), immunologic thrombocytopenic purpura (N = 3) and IgG multiple myeloma (N = 4). All control individuals presented low levels of red cell IgG (less than 38 IgG/RBC) with the exception of one of four patients with myeloma who had a mildly elevated value (50 IgG/RBC). Since the multiple myeloma patients had greater than 2 g/dl IgG, the possible nonspecific uptake of IgG onto the RBCs of patients with elevated serum IgG values did not interfere with the results of ELAT. ELAT proved to be a useful method for accurate quantification of the amount of IgG specifically bound on the surface of RBC of patients with AIHA.

Renal dysfunction in patients with sickle cell anemia or sickle cell trait.

Patients with sickle cell anemia (Hb SS) or sickle cell trait (Hb AS) may present several types of renal dysfunction; however, comparison of the prevalence of these abnormalities between these two groups and correlation with the duration of disease in a large number of patients have not been thoroughly investigated. In a cross-sectional study using immunoenzymometric assays to measure tubular proteinuria, microalbuminuria, measurement of creatinine clearance, urinary osmolality and analysis of urine sediment, we evaluated glomerular and tubular renal function in 106 adults and children with Hb SS (N = 66) or Hb AS (N = 40) with no renal failure (glomerular filtration rate (GFR) > 85 ml/min). The percentage of individuals with microalbuminuria was higher among Hb SS than among Hb AS patients (30 vs 8%, P < 0.0001). The prevalence of microhematuria was similar in both groups (26 vs 30%, respectively). Increased urinary levels of retinol-binding protein or beta 2-microglobulin were detected in only 3 Hb SS and 2 Hb AS patients. Urinary osmolality was reduced in patients with Hb SS or with Hb AS; however, it was particularly evident in Hb SS patients older than 15 years (median = 393 mOsm/kg, range = 366-469) compared with Hb AS patients (median = 541 mOsm/kg, range = 406-722). Thus, in addition to the frequently reported early reduction of urinary osmolality and increased GFR, nondysmorphic hematuria was found in 26 and 30% of patients with Hb SS or Hb AS, respectively. Microalbuminuria is an important marker of glomerular injury in patients with Hb SS and may also be demonstrated in some Hb AS individuals. Significant proximal tubular dysfunction is not a common feature in Hb SS and Hb AS population at this stage of the disease (i.e., GFR > 85 ml/min).

Treatment of T. cruzi infected human platelet concentrates with aminomethyltrimethyl psoralen (AMT) and ultraviolet A (UV-A) light: preliminary results.

The present measures adopted to prevent transfusion-associated Chagas' disease include screening of blood donors, and/or the inactivation of T. cruzi in collected blood using gentian violet (GV), as a trypanocidal agent. In this study, we investigated the efficacy of the combined use of AMT and UV-A in inactivating T. cruzi in infected human platelet concentrates. Human platelet concentrates were infected with T. cruzi (2 x 10(8)/ml) of the Y strain, transfered to PL 269 (Fenwal Laboratories) containers, and treated with GV (250 micrograms/ml), and ascorbic acid (1 mg/ml); GV, ascorbic acid and UV-A; GV and UV-A; AMT (40 microG/ml) and ascorbic acid; AMT, ascorbic acid and UV-A; AMT and UV-A; UV-A alone; and untreated (control). All UV-A treated platelet concentrates were exposed to UV-A doses of 24, 92, 184, 276, 368 and 644 kJ/m2, and the microscopical research of active T. cruzi was performed, using the microhematocrit technique, 1, 6 and 24 hours after each treatment. A high number of active forms of T. cruzi was observed in all condictions, except when GV was used as the trypanocidal agent, providing evidence of the failure of AMT and UV-A in inactivating T. cruzi in infected human platelet concentrates.