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Monteiro GC Hirata R Andrade AF Mattos-Guaraldi AL Nagao PE 《International journal of molecular medicine》2004,13(1):175-180
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. 相似文献
54.
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. 相似文献
55.
Claudio Gustavo Stefanoff Rocío Hassan Ana Carolina Gonzalez Luiz Ant?nio B Andrade Daniel G Tabak Sérgio Romano Ilana R Zalcberg 《Diagnostic molecular pathology》2003,12(2):79-87
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. 相似文献
56.
Structural response and relative strength of a laminated composite hip prosthesis: effects of functional activity 总被引:1,自引:0,他引:1
To obtain a better appreciation for the structural performance of a laminated composite hip prosthesis (CP), we examined in situ prosthesis structural response and relative strengths as a function of walking and stair climbing using our previously developed analysis guidelines. Accordingly, we examined overall prosthesis structural response utilizing a global continuum level modeling approach and prosthesis relative strengths using a local microstructural (or ply-level) modeling approach. As a reference and control, we examined the structural performance of the intact natural femur (NAT) and a titanium alloy (Ti) based hip prosthesis. In terms of the overall structural response, i.e., the femur/prosthesis deformational response, stem/bone interfacial stress transfer, and calcar strain energy density restored, the performance of the CP prosthesis was moderately improved over that of the control Ti prosthesis and better approximates the NAT response. In terms of relative strength, we found that the neck of the CP prosthesis failed for all activities with the exception of the mid-stance phase of level walking. However, the prosthesis appears to have sufficient relative strength for function at positions distal to the neck of the prosthesis. While these results dampen enthusiasm for consideration of laminated composite hip prostheses designed with a shape based on a metal alloy implant, they indirectly support consideration of alternate hip prosthesis structural designs such as using a better supported prosthesis neck or utilizing metal/composite hybrid constructions. Importantly, our simulation and analysis approach could be utilized in the design of other laminated composite biomedical structural components. 相似文献
57.
Maspin is expressed in the nuclei of breast myoepithelial cells 总被引:4,自引:0,他引:4
58.
Thalidomide, clinically used as an antiinflammatory and antitumoral drug, inhibited sponge-induced angiogenesis when administered systemically (100 mg/kg–1) in mice. However, it failed to inhibit solid Ehrlich tumor in the same mouse strain. We have used functional, biochemical and histological parameters to assess neovascularization and fibrovascular tissue infiltration of the mice sponge granuloma. The neovascularization growth as detected by development of blood flow and hemoglobin content extracted from the implants showed that thalidomide inhibited fibrovascular tissue formation by 40%. The functional and biochemical parameters correlated well with the histological study. Thalidomide had no inhibitory effect in the development of Ehrlich tumor. The detection of this selective action using the same animal strain bearing two different processes, supports the hypothesis that rather than species specificity, thalidomide is tissue specific. This approach may be used to identify the specificity of other therapeutic agents against distinct angiogenesis-dependent diseases. 相似文献
59.
Malaria, a major endemic tropical disease, is caused by the infection of blood cells by Plasmodium protozoa. Most patients control their parasitemia by a not fully understood spleen-dependent mechanism. SDF-1alpha is a chemokine produced by stromal cells such as reticular spleen cells. Nitric oxide (NO) has several immune functions, including killing of intracellular pathogens and its function in malaria is debated. We have previously shown that SDF-1alpha production peaks during the ascending parasitemia in Plasmodium chabaudi infection and its supplementation in lethal models could reduce the parasitemia. In the present study, we analyzed SDF-1 production by spleen cells as related to NO metabolism in the P. chabaudi rodent malaria model using IFN-gamma; TNFR and iNOS-knockout mice or iNOS-blocked, L-NAME- or aminoguanidine-treated mice. Parasitemia and production of SDF-1alpha and SDF-1beta were determined by RT-PCR. In vitro NO production by spleen adherent cells was also tested. The data showed that parasitemia was less intense in both iNOS(-/-) or NO-inhibited mice than in controls, with increased and long-lasting production of SDF-1alpha mRNA. In the absence of cytokines involved in the final regulation of NO production by effector cells, as is the case for TNFR(-/-) and GKO mice, the infection progressed in an uncontrolled manner regardless of SDF-1alpha production, suggesting that these cytokines must be involved in the control of parasitemia after the SDF-1alpha dependent process. The SDF-1beta isoform was constitutive in all experiments, with elevated levels only clearly seen in TNFR(-/-) mice. We conclude that SDF-1 is involved in the promotion of parasitemia control in malaria, and excessive NO could affect its production. 相似文献
60.
Ertekin-Taner N Ronald J Feuk L Prince J Tucker M Younkin L Hella M Jain S Hackett A Scanlin L Kelly J Kihiko-Ehman M Neltner M Hersh L Kindy M Markesbery W Hutton M de Andrade M Petersen RC Graff-Radford N Estus S Brookes AJ Younkin SG 《Human molecular genetics》2005,14(3):447-460
Plasma amyloid beta protein (Abeta42) levels and late onset Alzheimer's disease (LOAD) have been linked to the same region on chromosome 10q. The PLAU gene within this region encodes urokinase-type plasminogen activator, which converts plasminogen to plasmin. Abeta aggregates induce PLAU expression thereby increasing plasmin, which degrades both aggregated and non-aggregated forms of Abeta. We evaluated single nucleotide polymorphisms (SNPs) in PLAU for association with Abeta42 and LOAD. PLAU SNP compound genotypes composed of haplotype pairs showed significant association with AD in three independent case-control series. PLAU SNP haplotypes associated significantly with plasma Abeta42 in 10 extended LOAD families. One of the SNPs analyzed was a missense C/T polymorphism in exon 6 of PLAU (PLAU_1=rs2227564), which causes a proline to leucine change (P141L). We analyzed PLAU_1 for association with AD in six case-control series and 24 extended LOAD families. The CT and TT PLAU_1 genotypes showed association (P=0.05) with an overall estimated odds ratio of 1.2 (1.0-1.5). The CT and TT genotypes of PLAU_1 were also associated with significant age-dependent elevation of plasma Abeta42 in 24 extended LOAD families (P=0.0006). In knockout mice lacking the PLAU gene, plasma--but not brain--Abeta42 as well as Abeta40 was significantly elevated, also in an age-dependent manner. The PLAU_1 associations were independent of the associations we found among plasma Abeta42, LOAD and variants in the IDE or VR22 region. These results provide strong evidence that PLAU or a nearby gene is involved in the development of LOAD. PLAU_1 is a plausible pathogenic mutation that could act by increasing Abeta42, but additional biological experiments are required to show this definitively. 相似文献