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1.
Parasite Immunol ; 40(7): e12537, 2018 07.
Article in English | MEDLINE | ID: mdl-29777623

ABSTRACT

We aimed to investigate the association between plasma levels of complement Factor H (FH) with cardiac involvement, inflammatory and cardiometabolic parameters in patients with chronic Chagas' disease (CD). FH plasmatic levels were determined in 80 chronic CD patients. Glycaemic index, lipidogram (high-density lipoprotein cholesterol HDL-C, low-density lipoprotein cholesterol LDL-C, triglycerides and total cholesterol) and Ultrasensitive C-Reactive Protein (uCRP) values were obtained from medical records. Height, weight, body mass index (BMI) blood pressure and left ventricular ejection fraction (LVEF) were obtained from echocardiography examinations. Comparisons between chronic CD clinical forms were performed using Mann-Whitney test and correlation Spearman's test. FH levels were correlated positively with triglycerides (P = .001, r = .39), LDL-C (P = .009, r = .3), cholesterol (P = .02, r = .28), uCRP (P = .029, r = .31) and BMI (P = .008, r = .34); and negatively with HDL-C (P = .03, r = -.25) levels. Dyslipidemic patients showed higher FH levels compared to normolipidemic, although no difference for FH levels was observed between chronic CD clinical forms. Alternative pathway of complement may be a link between immune response and metabolic disorders, with important immunoregulatory role in chronic CD.


Subject(s)
Biomarkers/blood , Chagas Disease/immunology , Adult , Aged , Chronic Disease , Complement Factor H/analysis , Female , Humans , Male , Middle Aged
2.
Lupus ; 27(3): 372-379, 2018 Mar.
Article in English | MEDLINE | ID: mdl-28786770

ABSTRACT

The complement system contributes to the pathogenesis of systemic lupus erythematosus (SLE). Mannose-binding lectin (MBL) is a key molecule of the lectin pathway of complement and seems to be related to the clinical manifestations of this disease. We evaluated the serum levels of MBL and its relationship with disease onset and clinical findings in SLE patients. Serum samples were analysed in 195 patients and 145 healthy controls from southern Brazil. Patients with high MBL levels (above 2000 ng/ml) showed a significant increase in the frequency of thrombocytopaenia ( p = 0.007; OR = 2.71; 95% CI = 1.32-5.55); and seizures ( p = 0.034; OR = 2.61; 95% CI = 1.07-6.37). A positive correlation between disease activity and MBL levels (>2000 ng/ml; p = 0.031, rho = 0.279) as well as of MBL concentration with accumulated organ damage ( p = 0.021; rho = 0.232) was observed. Our results suggest a role for MBL in the development of clinical manifestations such as thrombocytopaenia and seizures in SLE patients. These findings corroborate the participation of the lectin pathway of complement in the pathophysiologic mechanisms underlying clinical manifestations of SLE.


Subject(s)
Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/complications , Mannose-Binding Lectin/blood , Seizures/blood , Thrombocytopenia/blood , Adult , Brazil , Case-Control Studies , Complement C3/metabolism , Complement C4/metabolism , Female , Humans , Logistic Models , Male , Mannose-Binding Lectin/genetics , Middle Aged , Seizures/etiology , Severity of Illness Index , Thrombocytopenia/etiology
3.
Cell Microbiol ; 19(4)2017 04.
Article in English | MEDLINE | ID: mdl-27665486

ABSTRACT

Extracellular vesicles released from pathogens may alter host cell functions. We previously demonstrated the involvement of host cell-derived microvesicles (MVs) during early interaction between Trypanosoma cruzi metacyclic trypomastigote (META) stage and THP-1 cells. Here, we aim to understand the contribution of different parasite stages and their extracellular vesicles in the interaction with host cells. First, we observed that infective host cell-derived trypomastigote (tissue culture-derived trypomastigote [TCT]), META, and noninfective epimastigote (EPI) stages were able to induce different levels of MV release from THP-1 cells; however, only META and TCT could increase host cell invasion. Fluorescence resonance energy transfer microscopy revealed that THP-1-derived MVs can fuse with parasite-derived MVs. Furthermore, MVs derived from the TCT-THP-1 interaction showed a higher fusogenic capacity than those from META- or EPI-THP-1 interaction. However, a higher presence of proteins from META (25%) than TCT (12%) or EPI (5%) was observed in MVs from parasite-THP-1 interaction, as determined by proteomics. Finally, sera from patients with chronic Chagas disease at the indeterminate or cardiac phase differentially recognized antigens in THP-1-derived MVs resulting only from interaction with infective stages. The understanding of intracellular trafficking and the effect of MVs modulating the immune system may provide important clues about Chagas disease pathophysiology.


Subject(s)
Cell-Derived Microparticles/metabolism , Chagas Disease/parasitology , Monocytes/parasitology , Trypanosoma cruzi/physiology , Animals , Antigens, Protozoan/immunology , Cell-Derived Microparticles/parasitology , Chagas Disease/immunology , Chagas Disease/metabolism , Chlorocebus aethiops , Host-Parasite Interactions , Humans , Membrane Fusion , Mice, Inbred BALB C , Monocytes/metabolism , Proteome/metabolism , Vero Cells
4.
Scand J Immunol ; 83(5): 314-20, 2016 May.
Article in English | MEDLINE | ID: mdl-26972106

ABSTRACT

Gum arabic and cashew nut tree gum exudate polysaccharide (CNTG) are plant polysaccharides composed of galactose and arabinose known as arabinogalactans (AGs). Although these fractions are used in food and pharmaceutical industry, cases of allergic reactions were described in clinical reports. As AGs were reported as modulators of the classical (CP) and alternative pathways (AP) of complement system (CS), in the present work, we investigate whether gum arabic and CNTG have an effect on both CS pathways. The complement fixation tests were performed with (CP-30 and AP-30) and without pre-incubation (CP-0 and AP-0). For CP-30, CNTG and gum arabic (833 µg/ml) showed a reduction of 28.0% (P = 0.000174) and 48.5% (P = 0.000143), respectively, on CP-induced haemolysis. However, no effect was observed for CP-0 in the CP-induced haemolysis. For AP-30, both CNTG and gum arabic (833 µg/ml) showed 87% reduction on the CP-induced haemolysis, with IC50 values of 100 and 7 µg/ml, respectively. For AP-0, a reduction of 11.3% for gum arabic and no effect for the CNTG on the CP-induced haemolysis were observed. These results suggested that gum arabic and CNTG could be acting as activators of the CS. Thus, this effect on the CS, especially on the AP, which accounts for up to 80-90% of total CS activation, indicates that both fractions may be harmful because of their potential pro-inflammatory action. Considering that CS activation induces inflammatory response, further studies confirming this immunomodulatory effect of these fractions are required to insure their safe use.


Subject(s)
Allergens/immunology , Complement Pathway, Alternative , Complement Pathway, Classical , Complement System Proteins/metabolism , Galactans/immunology , Hypersensitivity/immunology , Acacia/immunology , Anacardium/immunology , Animals , Cattle , Galactans/chemistry , Gum Arabic/chemistry , Hemolytic Plaque Technique , Humans , Rabbits
5.
Rev. bras. plantas med ; 18(1): 1-8, jan.-mar. 2016. tab
Article in Portuguese | LILACS | ID: lil-780051

ABSTRACT

RESUMO Verbena minutiflora Briq. ex Moldenke (gervai) tem seu uso medicinal relatado popularmente para tratamento de doenças hepáticas, diarreia e outros problemas de saúde. Entretanto, pouco se conhece a respeito de seus componentes químicos e estudos que comprovem suas propriedades medicinais são escassos. O objetivo desse estudo foi avaliar a composição química dos extratos aquosos e etanólicos de flores de V. minutiflora e otimizar processos de obtenção de extratos com maiores capacidades antioxidantes e maiores concentrações de flavonoides. O método de extração foi desenhado por planejamento fatorial, onde as variáveis para a determinação da capacidade antioxidante foram: pH, extração líquida, método e tempo de extração. Para a determinação de flavonoides totais as variáveis avaliadas por planejamento fatorial foram: concentração de hexametilenotetramina, tipo de ácido, volume de ácido e tempo de aquecimento. Os resultados das análises químicas dos extratos mostraram: aminogrupos, taninos e ácidos fixos (extrato aquoso) aminogrupos, flavonoides, triterpenos, esteroides, alcaloides e cumarinas (extrado hidroetanólico). Os resultados dos planejamentos fatoriais mostraram que o melhor método de extração para a capacidade antioxidante foi o que usou vórtex, por 35 min, com água:etanol 50:50, com pH1, obtendo 0,1899± 5,8.10-3 mmol expressos em ácido ascórbico g-1 nos extratos de V. minutiflora. Enquanto, para as dosagens de flavonoides totais as variáveis significantes foram: tipo de ácido e volume de ácido. A melhor extração obtida foi: 6,748. 10-2± 2,085 10-3% expressos em quercetina. Os resultados mostraram que o planejamento fatorial é uma importante ferramenta para a otimização de extração de componentes químicos em produtos naturais.


ABSTRACT Verbena minutiflora Briq. ex Moldenke (gervai) has its popular use reported for liver disorders treatments, diarrhea, and other health problems. However, little is known about its chemical components and studies that proves its medicinal properties are rare. The aim of this study was to evaluate the chemical composition of aqueous and ethanolic extracts from flowers of V. minutiflora and to optimize processes to obtain extracts with higher antioxidant capacity and greater concentration of flavonoids. The methods of extraction were designed by factorial planning, where the variables to determine the antioxidant capacity were: pH; extraction liquid; method and extraction time. To determinate the total flavonoids the variables evaluated by factorial design were: concentration of hexamethylenetetramine; type of acid; volume of acid and warming time. The results of chemical analysis of the extracts showed: amino groups, tannins and fixed acids (aqueous extract) amino groups, flavonoids, triterpenes, steroids, alkaloids and coumarins (hydroalcoholic extract). The factorial designs results showed that the best extraction method for the antioxidant capacity was the one that uses vortex, for 35 min, with water: ethanol 50:50, at pH 1, getting 0,1899 ± 5,8.10-3 mmol expressed in ascorbic acid g-1 in extracts of V. minutiflora . While, for dosages of total flavonoids the significant variables were the type of acid and volume of acid. The best extraction obtained was: 6,748. 10-2± 2,085 10-3% expressed in quercetin. These data showed that the factorial design is an important tool in optimizing the extraction of chemical components in natural products.


Subject(s)
/analysis , Chemistry , Verbena/chemistry , Process Optimization/classification , Pharmacognosy/methods , Factor Analysis, Statistical
6.
Int J Immunogenet ; 42(6): 439-44, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26385602

ABSTRACT

The aim of the study was to investigate the allotypic variability of complement factor B (BF) in patients and relatives with rheumatoid arthritis (RA) and its association with serological biomarkers and clinical features of the disease. BF allotypes were determined by high-voltage agarose gel electrophoresis in serum samples of 180 patients with RA, 198 relatives and 98 controls from Southern Brazil. Anticyclic citrullinated peptide (anti-CCP), antimutated citrullinated vimentin (anti-MCV) and IgA-rheumatoid factor (RF) were determined by ELISA and IgM-RF by latex agglutination in all samples. No significant differences were found in the allotypic variants of BF between patients with RA, relatives and controls, nor associations with gender and age of RA onset. BF*S07 allotype was significantly associated with extra-articular manifestations (EAMs; Secondary Sjögren Syndrome, pneumonitis, rheumatoid nodules) in patients with RA (P = 0.02; OR = 6.62). Patients with phenotype BF F had lower positivity for anti-MCV biomarker (P = 0.02; OR = 0.22) and those with allotype BF*S had higher prevalence of this autoantibody (P = 0.02; OR = 3.77). An increased frequency of RF-IgA was detected in relatives of patients with RA with BF FS07 phenotype (P = 0.02; OR = 7.78). Complement BF variability did not influence the development of RA in the studied patients, but BF variants may act as markers of disease prognosis, such as development of EAMs, corroborating with the role of the alternative pathway in the pathogenesis of RA.


Subject(s)
Arthritis, Rheumatoid/genetics , Arthritis, Rheumatoid/immunology , Autoantibodies/immunology , Complement Factor B/genetics , Complement Factor B/immunology , Family , Genetic Association Studies , Immunoglobulin Allotypes/immunology , Adult , Aged , Aged, 80 and over , Arthritis, Rheumatoid/blood , Autoantibodies/blood , Biomarkers , Brazil , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin Allotypes/blood , Male , Middle Aged , Odds Ratio , Phenotype , Young Adult
7.
Int J Immunogenet ; 41(3): 231-5, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24495190

ABSTRACT

Leprosy is one of the most neglected infectious tropical diseases of the skin and the nerves caused by the intracellular pathogen Mycobacterium leprae. The inducible NOS isoform encoded by NOS2A plays a vital role in host defence against bacterial infections. The functional promoter polymorphisms in NOS2A are associated with various autoimmune and infectious diseases. We investigated the association of NOS2A variants with progression of leprosy in a Brazilian cohort including 221 clinically classified patients and 103 unrelated healthy controls. We observed a novel variant ss528838018A/G in the promoter region at position -6558. The other functional variants were observed with low frequency of minor allele (<0.005). NOS2A promoter variant (-954G/C) was not observed in Brazilian populations, and the new observed promoter variant (ss528838018A/G) as well as other promoter variants were not associated with any clinical forms of leprosy in the Brazilian populations.


Subject(s)
Leprosy/genetics , Nitric Oxide Synthase Type II/genetics , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Adolescent , Adult , Aged , Aged, 80 and over , Alleles , Brazil , Female , Gene Frequency , Genotype , Humans , Leprosy/ethnology , Leprosy/microbiology , Male , Middle Aged , Mycobacterium leprae/physiology , Racial Groups
8.
Hum Immunol ; 72(9): 753-60, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21683108

ABSTRACT

Deficiency of mannan-binding lectin-associated serine protease 2 (MASP-2) has been associated with infections, whereas high levels appear to increase the risk of inflammatory disorders. Nevertheless, MASP2 haplotypes have been poorly investigated. To overcome haplotyping cost and time consumption, we developed multiplex polymerase chain reactions with sequence-specific primers (PCR-SSP) for 8 single nucleotide polymorphisms (SNPs), reducing the number of necessary reactions from 18 to 7. SNPs were distributed from the promoter to the last exon, and a single PCR-SSP was used for p.D120G. We evaluated the phylogenetic relationships and global distribution of 10 identified haplotypes in 338 Danish individuals with known MASP-2 and MAp19 levels and 309 South Brazilians. Four haplotypes were associated with reduced MASP-2 levels in plasma (lower than 200 ng/mL). Simultaneous association with the highest MASP-2 (over 600 ng/mL) and lowest MAp19 levels (lower than 200 ng/mL) was demonstrated with the intron 9 mutation (Kruskal-Wallis p < 0.0001). Cumulative genotype frequencies predict approximately 0.4% severely deficient and 25% overproducing individuals in both populations. Rapid and low-cost screening of patients with multiplex MASP2 PCR-SSP could be used to identify clinical conditions where MASP-2 (or MAp19) levels may be disease modifying, possibly improving disease outcome through early therapeutic and preventive measures.


Subject(s)
Autoimmune Diseases/genetics , Ethnicity , Infections/genetics , Mannose-Binding Protein-Associated Serine Proteases/metabolism , Alternative Splicing/genetics , Biomarkers/metabolism , Brazil/ethnology , Denmark/epidemiology , Gene Frequency , Genetic Predisposition to Disease , Haplotypes , High-Throughput Screening Assays , Humans , Mannose-Binding Protein-Associated Serine Proteases/genetics , Multiplex Polymerase Chain Reaction , Phylogeny , Polymorphism, Single Nucleotide
9.
Clin Exp Immunol ; 157(3): 395-9, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19664148

ABSTRACT

Ficolins are pattern-recognition proteins involved in innate immunity, which upon binding to their specific pathogen-associated molecular patterns on the microbial surfaces trigger the immune response either by binding to collectin cellular receptors or by initiating the complement lectin pathway. In humans, three ficolin genes have been identified, which encode ficolin-1 (M-ficolin), ficolin-2 (L-ficolin) and ficolin-3 (H-ficolin or Hakata antigen). Ficolin-2 was shown to bind to lipoteichoic acid, a cell wall constituent in all Gram-positive bacteria such as Streptococcus pyogenes, which is the aetiological agent of rheumatic fever (RF) and its most severe sequelae, chronic rheumatic heart disease (CRHD). Here we investigated polymorphisms in the promoter region of the FCN2 gene (at positions -986/-602 and +4) in 122 patients with RF and CRHD and in 210 healthy subjects from the same geographic region and socioeconomic background. The haplotype -986/-602/-4 G/G/A, which is related to low levels of L-ficolin, was observed more frequently in the CRHD group when compared to the healthy subjects [99/162, 61.1% versus 211/420, 50.2%, odds ratio (OR) 1.6, confidence interval (CI) 95% 1.1-2.3, P = 0.021]. The haplotype -986/-602/-4 A/G/A was observed more frequently in the healthy group when compared to the affected (RF plus CRHD) subjects (31/420, 7.4% versus 6/244, 2.5%, OR 3.2, CI 95% 0.13-0.77, P = 0.008). Based on those findings, one can conclude that polymorphisms associated with low levels of L-ficolin level may predispose an individual to recurrent and/or more severe streptococcal infection.


Subject(s)
Lectins/genetics , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Rheumatic Fever/genetics , Streptococcal Infections/genetics , Adolescent , Adult , Aged , Case-Control Studies , Child , Chronic Disease , Female , Genetic Predisposition to Disease , Haplotypes , Humans , Lectins/blood , Lectins/deficiency , Logistic Models , Male , Middle Aged , Rheumatic Heart Disease/genetics , Risk , Young Adult , Ficolins
10.
Int J Cardiol ; 109(2): 275-6, 2006 May 10.
Article in English | MEDLINE | ID: mdl-15946754

ABSTRACT

Chronic rheumatic valve disease (CRVD) is a late sequel of Rheumatic Fever (RF) which appears in approximately 30% of RF patients, leading to valve injury. Advanced Oxidation Protein Products (AOPP) and high sensitive C-Reactive Protein (hs-CRP) plasma levels were measured in patients with CRVD in order to evaluate the presence of oxidative stress and systemic inflammation. A total of 90 patients (70 female, 20 male, mean age 46.01 +/- 11.72 years, range 24-69 years) with CRVD, who have or have not undergone valve replacement due to rheumatic etiology, and 46 healthy subjects (27 female, 19 male, mean age 41.89 +/- 9.02 years range 28-60 years) were studied. Levels of AOPP were measured by the determination of optical density (OD) at 340 nm under acidic conditions and hs-CRP by enhanced immunonephelometric assays. Significantly elevated levels of AOPP and hs-CRP were observed in CRVD patients when compared to the controls (AOPP 212.62 +/- 34.14 umol/l vs. 126.97 +/- 27.74 umol/l p < 0.00006 and for hs-CRP 5.40 +/- 1.98 mg/l vs. 2.66 +/- 1.36 mg/l p < 0.05). In addition, high levels of AOPP were associated to the presence of prosthetic valve and time after surgery (p < 0.0008 and p < 0.005, respectively). No correlation was observed between the levels of AOPP and hs-CRP with age, sex and degree of mitral valve stenosis. No correlation was found between AOPP and hs-CRP plasma values. These results suggest the involvement of oxidative stress and systemic inflammation in the pathogenesis of CRVD.


Subject(s)
C-Reactive Protein/metabolism , Inflammation Mediators/blood , Mitral Valve Stenosis/blood , Oxidative Stress , Rheumatic Heart Disease/blood , Adult , Aged , Brazil/epidemiology , Case-Control Studies , Chronic Disease , Female , Heart Valve Prosthesis Implantation , Humans , Male , Middle Aged , Mitral Valve Stenosis/physiopathology , Mitral Valve Stenosis/surgery , Rheumatic Heart Disease/physiopathology , Rheumatic Heart Disease/surgery , Treatment Outcome
11.
Braz. j. infect. dis ; 9(6): 459-463, Dec. 2005. tab
Article in English | LILACS | ID: lil-419677

ABSTRACT

The use of highly active antiretroviral therapy (HAART) for the treatment of HIV infection has been associated with a marked reduction in the incidence of most opportunistic infections. From April 2001 to February 2002, 80 blood samples from patients who were suspected to have disseminated mycobacterial infection, presenting fever and (preferably) a CD4 T cell count < 100.0 cell/mL were investigated. Twelve (15 percent) of the 80 blood cultures were positive for mycobacteria, with Mycobacterium avium being identified in 7 (8.8 percent) samples and M. tuberculosis in 5 (6.2 percent). The TCD4+ count at the time of M. avium bacteremia ranged from 7cells/æL (average of 48.5 cell/æL), while in M. tuberculosis bacteremia it ranged from 50.0 cells/æL (average of 80.0 cell/æL). The prevalence of M. avium bacteremia in our study follows the expected decline in opportunistic infections observed after the introduction of HAART; however, mycobacteremia by M. tuberculosis still indicates a high prevalence of tuberculosis infection in AIDS patients.


Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , AIDS-Related Opportunistic Infections/epidemiology , Mycobacterium avium Complex/isolation & purification , Mycobacterium avium-intracellulare Infection/epidemiology , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Pulmonary/epidemiology , AIDS-Related Opportunistic Infections/diagnosis , AIDS-Related Opportunistic Infections/microbiology , Antiretroviral Therapy, Highly Active , Brazil/epidemiology , Mycobacterium avium-intracellulare Infection/diagnosis , Mycobacterium avium-intracellulare Infection/microbiology , Prevalence , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/microbiology
12.
Parasite Immunol ; 27(9): 333-40, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16149991

ABSTRACT

The activation of complement on the surface of Leishmania promastigotes appears to be an important factor for parasite infectivity in the mammalian host, allowing their attachment and the invasion of macrophages via complement receptors. Mannose-binding lectin (MBL) is a well-known complement activator and an efficient opsonine. We have investigated here whether serum and purified MBL bind to and promote lysis of live promastigotes of L. braziliensis; and evaluated the deposition of MBL, C1q, C4 and C3 on the parasite surface after interaction with non-immune normal human serum (NHS). We observed that both serum MBL and the purified MBL-MASP complex bind to the surface of L. braziliensis and that this binding occurred via the carbohydrate recognition domains of MBL. The binding of MBL, however, did not affect the lytic effect of complement on the parasites. The deposition of C1q, C4, C3 and parasite lysis was observed after incubation with NHS. EDTA but not EGTA abolished C3 deposition on the parasite surface, indicating the involvement of the alternative pathway in this process. Our results indicate that MBL binds to L. braziliensis and that this is mediated by a specific carbohydrate on the surface of parasites and provides evidence for antibody-independent mechanisms that complement activation on the parasite surface.


Subject(s)
Complement Activation , Leishmania braziliensis/immunology , Mannose-Binding Lectin/immunology , Mannose-Binding Lectin/metabolism , Membrane Glycoproteins/immunology , Membrane Glycoproteins/metabolism , Animals , Complement C1q/metabolism , Complement C3/metabolism , Complement C4 , Cytotoxicity, Immunologic , Humans , Immunohistochemistry , Microscopy, Confocal , Protein Binding , Protein Interaction Mapping , Protein Structure, Tertiary , Protozoan Proteins/immunology , Protozoan Proteins/metabolism
13.
Braz J Infect Dis ; 9(6): 459-63, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16410940

ABSTRACT

The use of highly active antiretroviral therapy (HAART) for the treatment of HIV infection has been associated with a marked reduction in the incidence of most opportunistic infections. From April 2001 to February 2002, 80 blood samples from patients who were suspected to have disseminated mycobacterial infection, presenting fever and (preferably) a CD4 T cell count < 100.0 cell/mL were investigated. Twelve (15%) of the 80 blood cultures were positive for mycobacteria, with Mycobacterium avium being identified in 7 (8.8%) samples and M. tuberculosis in 5 (6.2%). The TCD4+ count at the time of M. avium bacteremia ranged from 7 cells/microL (average of 48.5 cell/microL), while in M. tuberculosis bacteremia it ranged from 50.0 cells/microL (average of 80.0 cell/microL). The prevalence of M. avium bacteremia in our study follows the expected decline in opportunistic infections observed after the introduction of HAART; however, mycobacteremia by M. tuberculosis still indicates a high prevalence of tuberculosis infection in AIDS patients.


Subject(s)
AIDS-Related Opportunistic Infections/epidemiology , Mycobacterium avium Complex/isolation & purification , Mycobacterium avium-intracellulare Infection/epidemiology , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Pulmonary/epidemiology , AIDS-Related Opportunistic Infections/diagnosis , AIDS-Related Opportunistic Infections/microbiology , Adolescent , Adult , Antiretroviral Therapy, Highly Active , Brazil/epidemiology , CD4 Lymphocyte Count , Child , Female , Humans , Male , Middle Aged , Mycobacterium avium-intracellulare Infection/diagnosis , Mycobacterium avium-intracellulare Infection/microbiology , Prevalence , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/microbiology
14.
Eur J Clin Microbiol Infect Dis ; 23(11): 851-4, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15558344

ABSTRACT

The study presented here evaluated the utility of several methods of extracting mycobacterial nucleic acids from positive blood culture samples and examined the effect of each method on the performance of an in-house PCR used directly in the peripheral blood of 80 patients with AIDS to identify Mycobacterium spp. The modified Boom method for extracting DNA from blood cultures proved to be the most efficient, with subsequent PCR analysis yielding 100% positivity (7 samples positive for M. avium and 5 for M. tuberculosis). Only three of 12 patients with a positive blood culture had a PCR result positive for M. avium in peripheral blood. The identification of mycobacteria by PCR in blood culture took about 3 days, reducing the time to diagnosis by several weeks. These results demonstrate that PCR is a sensitive and quick method for identifying mycobacteria, especially when a good DNA extraction method is applied.


Subject(s)
DNA, Bacterial/isolation & purification , Mycobacterium avium Complex/isolation & purification , Mycobacterium tuberculosis/isolation & purification , Acquired Immunodeficiency Syndrome/complications , Adolescent , Adult , Child , DNA, Bacterial/blood , Female , Humans , Male , Middle Aged , Mycobacterium Infections/diagnosis , Polymerase Chain Reaction , Sensitivity and Specificity
15.
Clin Exp Immunol ; 138(3): 521-5, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15544631

ABSTRACT

Although mannose-binding lectin (MBL) is known to be involved in the primary defense against microorganisms, there are emerging lines of evidence for an active proinflammatory role for MBL in different chronic diseases. In this study we determined the circulating levels of MBL in patients with rheumatic heart disease (RHD). A total of 100 patients (77 women, 23 men; mean age 45.8 +/- 11 years, range 19-76 years) with chronic RHD, and a previous diagnosis of rheumatic fever, were studied. Transthoracic echocardiography was performed in all patients to evaluate valvular heart disease. Ninety-nine healthy individuals matched for age, sex and ethnic origin were included as controls. MBL concentration was measured by enzyme-linked immunosorbent assay and C3 and C4 levels by turbidimetry. MBL levels were significantly higher in patients with RHD than in healthy subjects (mean +/- SEM: 3036.2 +/- 298.9 ng/ml versus 1942.6 +/- 185.5 ng/ml, P <0.003). In addition, MBL deficiency was more prevalent in controls (17.1%) than in patients (9% P <0.09). Concentrations of C4 were within the normal range (22.7 +/- 0.8 mg/dl, normal: 10.0-40.0 mg/dl), while C3 concentrations were found to be elevated (109.2 +/- 3.6 mg/dl, normal: 50.0-90.0 mg/dl). No correlation was observed between serum MBL levels and valve area or the type of surgical procedure. The significantly elevated circulating MBL levels in patients with RHD together with the greater prevalence of MBL deficiency in controls suggest that MBL may cause undesirable complement activation contributing to the pathogenesis of RHD.


Subject(s)
Mannose-Binding Lectin/blood , Rheumatic Heart Disease/blood , Adult , Aged , Complement C3/analysis , Complement C4/analysis , Female , Humans , Male , Mannose-Binding Lectin/deficiency , Mannose-Binding Lectin/immunology , Middle Aged , Rheumatic Heart Disease/immunology , Rheumatic Heart Disease/surgery
16.
Tissue Antigens ; 62(4): 308-12, 2003 Oct.
Article in English | MEDLINE | ID: mdl-12974797

ABSTRACT

The heterogeneity in the clinical expression of Chagas disease gives strong evidences for the involvement of genetic factors on its pathogenesis. Several studies have indicated different markers of genetic susceptibility to Chagas cardiomyopathy. In the present study, we present evidence of association between complement C3 and BF allotypes, and the susceptibility to Chagas disease and the development of cardiomyopathy. C3, BF, C4A, C4B and C2 polymorphism were determined in 100 seropositive Chagasic patients [cardiomyopathic (CARD), n = 57; asymptomatic indetermined (IND), n = 43] and in 100 non-related seronegative healthy controls. Patients and controls were matched according to their ethnic and geographical origin. A significantly increased frequency of C3F was observed in patients with the CARD form (8/57 14.03%), when compared with those presenting the IND form (0/43, 0%; RR 7.0) and with the healthy controls (5/100, 5%; RR 3.1). A negative association of the BF S allotype was observed in the CARD patients (19/57 33.33%) and in the Chagas total (38/100 38.0%), when compared with the controls (55/100, 55.0%; RR 0.4). All other C3, BF, C4A, C4B and C2 alleles showed no significant differences. These results suggest the allele C3F as a susceptible marker for the progression of the CARD form. On the other hand, BF S may represent a protective role against severe CARD disease. These results corroborate the importance of the alternative pathway in Trypanosoma cruzi infection and indicate possible genetic markers of Chagas cardiomyopathy.


Subject(s)
Chagas Cardiomyopathy/genetics , Complement C3/genetics , Genetic Predisposition to Disease , Adult , Aged , Blood Group Antigens/genetics , Chagas Cardiomyopathy/blood , Chagas Cardiomyopathy/immunology , Complement Activation , Complement Factor B/genetics , Female , Humans , Immunoglobulin Allotypes/genetics , Male , Middle Aged , Myocardium/pathology , Polymorphism, Genetic
17.
Clin Exp Immunol ; 127(2): 310-5, 2002 Feb.
Article in English | MEDLINE | ID: mdl-11876755

ABSTRACT

In an infectious process complement activation is necessary for a proper immune and inflammatory response, but when exacerbated may cause tissue injuries. In infective endocarditis (IE) patients tend to develop high titres of circulating immune complexes (CIC) that activate complement. The aim of this study was to evaluate for the first time complement activation in IE for possible correlation with extracardiac manifestations and clinical prognosis. Twenty patients with IE, 14 healthy controls and 15 patients presenting mitral and aortic valve lesions (with no signs of either infection or other associated diseases), were studied. Plasma levels of C3adesArg, SC5b-9, C1rs-C1Inh and C3b(Bb)P were determined by ELISA and C3d by double decker immunoelectrophoresis. C3 and C4 levels were assayed by turbidimetry and CIC by ELISA. Elevation of plasma levels of all complement activation products, with the exception of C3b(Bb)P, indicated a significant classical pathway activation in IE patients when compared to controls (C3d: P < 0.00004; C3adesArg: P < 0.03, SC5b-9: P < 0.01, C1rs-C1Inh: P < 0.00007). CIC levels were significantly increased (P < 0.005) and C3 reduced in IE patients (P < 0.05). Elevated C3d (P < 0.02) and C3adesArg (P < 0.03) levels were associated with pulmonary manifestations. In addition, C3d was significantly elevated in the patients who died when compared to those who had a good recovery (P < 0.02). Our data demonstrate the activation of the complement classical pathway, most probably mediated by CIC, in IE and suggests C3d and C3adesArg as possible markers for extracardiac lesion and severity of the disease.


Subject(s)
Complement Activation , Endocarditis, Bacterial/immunology , Adolescent , Adult , Aged , Antigen-Antibody Complex/blood , Bacteremia/complications , Central Nervous System Diseases/etiology , Complement C3d/analysis , Complement Membrane Attack Complex/analysis , Endocarditis, Bacterial/blood , Endocarditis, Bacterial/complications , Endocarditis, Bacterial/mortality , Female , Humans , Kidney Diseases/etiology , Lung Diseases/etiology , Male , Middle Aged , Prognosis , Rheumatoid Factor/blood , Splenomegaly/etiology
18.
Dig Dis Sci ; 46(12): 2624-30, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11768251

ABSTRACT

The coexistence of celiac disease together with a range of autoimmune disorders has already been reported. The aims of this study were to perform a broad spectrum of autoantibodies in celiac patients (N = 56), their first-degree relatives (N = 118), and compare the data with healthy controls (N = 101) and patients with inflammatory bowel disease (N = 42; Crohn's disease, N = 18 and ulcerative colitis, N = 24). All serum samples were tested by indirect immunofluorescence to the anti-endomysium antibodies (EmA), anti-neutrophil cytoplasmic (ANCA), anti-smooth-muscle (SMA), anti-mitochondrial (AMA), anti-nuclear (ANA), anti-liver-kidney microsomal (LKM), anti-gastric parietal cells (GPCA), and anti-thyroid microsome (TMA). EmA were detected in 100% of celiac patients ingesting gluten and in 16.1% of the first-degree relatives, while ANCA were positive only in patients with ulcerative colitis (45.6%) and Crohn's disease (16.5%). Fourteen CD patients (25%) were positive for at least one of the other autoantibodies, with significant prevalence of TMA, ANA, and GPCA, while the relatives showed 17.8% of positivity, with an increased prevalence of ANA and TMA. These results emphasize the value of screening for different autoantibodies in celiac patients and their relatives and corroborate the need for evaluation and follow-up of these individuals.


Subject(s)
Autoantibodies/blood , Celiac Disease/immunology , Colitis, Ulcerative/immunology , Crohn Disease/immunology , Adolescent , Adult , Aged , Antibodies, Antineutrophil Cytoplasmic/blood , Child , Child, Preschool , Female , Fluorescent Antibody Technique, Indirect , Humans , Male , Middle Aged
19.
J Investig Allergol Clin Immunol ; 10(4): 209-14, 2000.
Article in English | MEDLINE | ID: mdl-11039837

ABSTRACT

The aim of this study was to evaluate the complement activation in Brazilian patients with preeclampsia and to correlate it with the severity and clinical outcome of the disease. Plasma levels of C3d, SC5b-9, C3 and C4 were measured in 16 patients with preeclampsia and in 17 normotensive pregnant women. Ten patients developed severe and six mild disease. C3 and C4 levels were determined by turbidimetry using polyclonal specific antisera. C3d concentrations were evaluated through double-decker rocket immunoelectrophoresis and SC5b-9 was assayed by a double-antibody enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody against a neoantigen expressed in the formed complex. The mean levels of all variables were significantly higher in the preeclamptic group (before the delivery) when compared to the normal pregnancies. The complex SC5b-9 followed by C3d showed the most significant results for those comparisons (p < or = 0.00001). The levels of all parameters in the preeclampsia patients decreased significantly after the delivery. Again, the complex SC5b-9 and C3d showed the most significant results (p < or = 0.0004). None of the studied variables showed statistically significant differences regarding the severity of preeclampsia. These results confirm the activation of complement in preeclampsia, suggesting that this activation is related to the disease manifestation. Our findings further emphasize the involvement of complement activation in the pathological manifestations of preeclampsia.


Subject(s)
Complement Activation , Pre-Eclampsia/immunology , Adolescent , Adult , Brazil , Complement Activation/immunology , Complement System Proteins/analysis , Female , Humans , Pregnancy , Severity of Illness Index
20.
Rev Panam Salud Publica ; 7(6): 371-6, 2000 Jun.
Article in Portuguese | MEDLINE | ID: mdl-10949897

ABSTRACT

This study investigated the autoantibody profile of 241 blood samples from 176 Kaingang and 65 Guarani Indians from three populations living on the Rio das Cobras and Ivaí reservations, in the state of Paraná, in southern Brazil. The presence of antimitochondrial, anti-smooth muscle, antinuclear, anti-parietal cell, and anti-liver-kidney microsome antibodies was determined by indirect immunofluorescence. These results were compared with samples from 100 healthy Caucasian individuals from the general population of the state. Total positivity was 9% for the indigenous population and 4% for the control population. The prevalence of anti-smooth muscle antibodies was significantly higher among the Guarani and Kaingang individuals from the Rio das Cobras reservation (P = 0.03). It is likely that the increased exposure that these indigenous Brazilians have to infectious diseases that were previously unknown to them comes from more contact with non-native populations, growing acculturation, and cultural practices that include scarification and tattooing. The presence of auto-antibodies in these Brazilian Indians may be related to mechanisms of molecular mimicry with viral or bacterial antigens.


Subject(s)
Autoantibodies/analysis , Indians, South American , Adolescent , Adult , Antibodies, Antinuclear/analysis , Brazil , Female , Fluorescent Antibody Technique, Indirect , Humans , Male , Middle Aged
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