Interleukin 10 protects mice against staphylococcal enterotoxin B-induced lethal shock.

We investigated the ability of interleukin 10 (IL-10) to protect mice against lethal shock induced by staphylococcal enterotoxin B (SEB). Treatment of mice with IL-10 prevented the death of mice injected with SEB in a dose-dependent manner. IL-10-mediated protection was apparent when administered either prior to or concurrent with SEB but was less effective when administered following SEB injection. This observation indicates that IL-10 is capable of regulating T-cell activation in vivo.     

Housing and house-dust mites

Because the mite-allergen content in homes is highly variable even in the same geographic area, we tried to determine which variables influence mite infestation. We evaluated mite-allergen content in bedding relative to housing conditions and living habits. This cross-sectional study included 108 homes. Housing conditions were assessed by an architect and living habits by a researcher specialized in social and family economics. Group I allergen level was measured on the mattress dust with monoclonal antibodies, and relative humidity (RH) was monitored in the bedroom during a 2-week period. Homes with low RH did have low mite-allergen content. In contrast, homes with intermediate RH levels had very variable mite-allergen content. Using analysis of variance and a logistic regression analysis, we were unable to identify any variable predictive of mite-allergen content. Thus, factors other than relative humidity seem to influence mite infestation. Moreover, the absence of association between mite infestation and ventilation rate might be accounted for by the gentle climate in our area with notable outdoor RH.     

Cytogenetic analysis of 100 consecutive newly diagnosed cases of acute lymphoblastic leukemia in Rio de Janeiro

We report the cytogenetic analysis of newly diagnosed Brazilian children with acute lymphocytic leukemia (ALL). We investigated 100 ALL cases from four different institutions in Rio de Janeiro. The frequency of chromosomal abnormalities was 92.3%. The karyotype profile and recurrent abnormalities found in this study do not differ essentially from those described by other groups. Although the Brazilian population is usually the product of different ethnic groups, our results show that the frequency of each recurrent abnormality is similar to that found in populations without our degree of diverse ethnic composition. Hence, our results suggest that childhood ALL in Brazil has the same biological features as that in developed countries, supporting the use of similar treatment protocols. We can therefore expect to reach the same survival rates in the coming years, depending possibly on the efficacy of the support therapy and extent of social assistance.     

Bone marrow transplantation for diamond-blackfan anemia.

Patients with Diamond-Blackfan anemia (DBA) who are unresponsive to or intolerant of corticosteroids, experience treatment failure with other treatments, develop additional cytopenias or clonal disease, or opt for curative therapy are often treated with allogeneic bone marrow transplantation. We studied the transplantation outcomes of 61 DBA patients whose data were reported to the International Bone Marrow Transplant Registry between 1984 and 2000. The median age was 7 years (range, 1-32 years). Among 55 patients with available transfusion information, 35 (64%) had received > or =20 units of blood before transplantation. Most patients (67%) received their bone marrow grafts from an HLA-matched related donor. The median time to neutrophil recovery was 17 days (range, 10-119 days) and to platelet recovery was 23 days (range, 9-119 days). Five patients did not achieve neutrophil engraftment. The 100-day mortality was 18% (95% confidence interval, 10%-29%). Grade II to IV acute graft-versus-host disease occurred in 28% (range, 17%-39%) and chronic graft-versus-host disease in 26% (range, 15%-39%). The 3-year probability of overall survival was 64% (range, 50%-74%). In univariate analysis, a Karnofsky score > or =90 and transplantation from an HLA-identical sibling donor were associated with better survival. These data suggest that allogeneic bone marrow transplantation is effective for the treatment of DBA. Transplantation before deterioration of the performance status and from an HLA-identical sibling donor may improve survival.     

Surface carbohydrates as recognition determinants in non-opsonic interactions and intracellular viability of group B Streptococcus strains in murine macrophages

Mononuclear cells have been found to play a key role in phagocytosis and eventual killing of group B streptococci (GBS). The rich array of sugars on bacterial surface plus the presence of membrane-associated lectin-receptors on the macrophage suggests that this is a likely means for GBS recognition by these host defense cells. Macrophages have been shown to bind GBS in the absence of serum components. However, participation of carbohydrate moieties in GBS intracellular survival had not been completely elucidated. The aim of this study was to assess the involvement of sugars on adherence and intracellular viability in murine macrophages of GBS serotypes Ia (85147 and 90222 strains), III (80340 and 90356 strains) and V (88641 and 90186 strains) isolated from assymptomatic carriers and patients, respectively. Most isolates showed higher adherence within 2-h incubation. Only 90222-Ia strain exhibited progressive adherence rate until 12-h incubation. All strains showed intracellular viability during first 0.5-h of incubation. Except for 90186-V strain that survived only for 2 h, strains of all serotypes tested were found to survive 24 h into macrophages. Treatments of bacteria by glycosidases inhibited macrophage interaction with GBS strains at varied levels. Neuraminidase inhibited 90-97% adherence and 100% intracellular survival of GBS strains (P<0.0001). Host cell treatments with Rhamnose, N-acetyl-D-glucosaminidase and Fucose (5 mg/ml) inhibited adherence and intracellular viability of GBS strains at varied levels. Removal of GlcNAc residues of invasive GBS isolates enhanced intracellular viability, suggesting that GlcNAc residues may act by intercepting the expression of hidden receptors probably related with invasiveness and survival within macrophages. Lastly, our results demonstrate involvement of sialic acid specific receptors on macrophages and lectinophagocytosis in non-opsonic interaction and survival of GBS invasive isolates.     

On-line sensors for coagulation proteins: concept and progress report

The assessment of blood damage and of the activation of the coagulation, complement and/or inflammatory systems by cardiovascular and extracorporeal devices is difficult at best. Immunoassay methods are now available for the measurement of many of the proteins, enzymes and peptides involved in coagulation, thrombosis, complement and inflammation. We present a long-range project and plan to develop an array of remote, on-line, semicontinuous immunosensors for selected coagulation proteins, based on fluoroimmunoassay principles. The free/bound separation step is performed optically. Excitation of fluorescence is performed via an evanescent wave produced by total internal reflection and waveguide optics. Fluorescence emission is collected only in the near field. Means to deliver fluorescently-labelled reagent and to modify the antigen-antibody binding constant are presented and discussed. The results of non-specific binding, plasma-blood fluorescence, and blood compatibility are also discussed.     

Notes on parasitism by Amblyomma humerale (Acari: Ixodidae) in the state of Rondônia,western Amazon,Brazil

The tick Amblyomma humerale Koch is endemic to South America. All host records refer to the adult stage parasitizing tortoises, mostly yellow-footed tortoise, Geochelone denticulata (L.), and red-footed tortoise, Geochelone carbonaria (Spix). The current study reports the presence of A. humerale in the state of Rond?nia, Brazil. A total of 215 adult ticks (201 males, 14 females) was collected from six G denticulata in an Indian reserve and nine Geochelone sp. in rural Monte Negro County, giving an overall mean infestation of 14.3 +/- 12.0 (range: 2-44) ticks per tortoise. Male ticks always outnumbered females on the host and nine tortoises had only male ticks. Male ticks were mostly attached in clusters on the ventral sides of the carapace near the anterior and posterior margins, and more rarely on the outer margin of the plastron. All females were found attached to the tortoise skin, at different sites such as head, neck, shoulders or legs. Male ticks were rarely observed attached to the body skin. Seven engorged nymphs collected on small vertebrates from Monte Negro County molted to adults of A. humerale. This included one nymph each on the seven-colored lizard, Plica plica (L), green tree climber, Plica umbra (L.), and wide-foraging lizard, Kentropyx calcarata Spix,three nymphs on the common opossum, Didelphis marsupialis L., and one nymph on the silky anteater, Cyclopes didactylus L. These constitute the first host records for the immature stages of the tick A. humerale.     

Laboratory strategies for efficient handling of paraffin-embedded tissues for molecular detection of clonality in non-hodgkin lymphomas.

We herein present a technical strategy to optimize DNA isolation from paraffin-embedded tissue (PET). This includes the choice of adequate buffers for proteinase K digestion and multiplex PCR amplifications for assessing the appropriateness of DNA extracts for subsequent PCR assays for detecting clonality. We found that the association of proteinase K digestion in nonionic buffer and subsequent extract dilutions accounted for 79% of successful amplifications. A final efficiency of 88% was achieved by additional organic extractions and/or re-extractions. Comparisons were carried out with control DNA extracts from fresh samples to assess the efficiency of each clonality assay. Immunoglobulin CDRIII rearranged region amplification was more efficient for pregerminal center B-cell lymphomas in contrast to CDRII rearrangement detection, which was more effective for germinal and postgerminal lymphomas. T-cell clonality detection by TCRgamma PCR was less efficient in PET samples than in fresh tissues showing that DNA integrity is more critical for TCR than for IGH amplification. Two inconclusive cases without phenotypic markers and two other atypical lymphoproliferations masked by reactive T cells were diagnosed as plasmablastic lymphomas and as monoclonal B-proliferations, respectively, due to IGH rearrangements.