The role of cholinesterases in chagas disease.

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) hydrolyze acetylcholine (ACh) in neural synaptic clefts and are primarily found in erythrocytes and blood plasma, respectively. Besides inactivating ACh, cholinesterases may play a non-classical role in inflammation and in immune response. In a previous study, we reported that BChE levels were decreased in chronic Chagas disease patients presenting the mega syndromes. In this series, we reported that: i) the activity of AChE did not differ between patients and controls, irrespective of the presence or not of the 1057C > A ACHE polymorphism, and ii) the increased BChE levels modestly influenced the AChE activity in Chagas disease.

Water quality assessment of the Demetrio stream: an affluent of the Gravataí River in the South of Brazil / Qualidade da água do arroio Demetrio: um afluente do rio Gravataí no sul do Brasil

The Gravataí River basin, one of the main water sources of the metropolitan region of Porto Alegre, is among the ten most polluted rivers in Brazil. Water quality is monitored only through physico-chemical and microbiological parameters in Brazil, and in this context, considering the importance of the use of biomarkers in complementing the analysis of water, the present study aimed to evaluate the environmental quality of the main affluent of the Gravataí River, Demetrio stream, through physico-chemical, microbiological, and cytogenotoxic criteria, at the stream source (P1), whereas samples P2 and P3 were obtained from the upstream near the area with the highest urban density and the downstream near the meeting point with the Gravataí River, respectively. The results for copper concentration and color classified the Demetrio stream as Class 4 in general, that is, the water is suitable only for navigation and to landscape harmony. The main genotoxic alterations (micronuclei and nuclear buds) were observed in P2, in which were obtained the highest levels of copper, in addition to iron and manganese. Anthropic interventions were observed in P1 and P2; however, due to its low metal concentration, P3, near the Gravataí River, manifested an improvement in environmental quality.

A bacia do rio Gravataí, uma das principais fontes de água da região metropolitana de Porto Alegre, está entre os dez rios mais poluídos do Brasil. No Brasil a qualidade da água é monitorada apenas através de parâmetros físico-químicos e microbiológicos e, nesse contexto, considerando a importância do uso de biomarcadores para complementar a análise da água, o presente estudo teve como objetivo avaliar a qualidade ambiental do principal afluente do Rio Gravataí, o arroio Demétrio, através de critérios físico-químicos, microbiológicos e citogenotóxicos, na nascente do arroio (P1), a montante e próximo à área com maior densidade urbana (P2) e a jusante e próximo ao ponto de encontro com o rio Gravataí (P3). Os resultados para a cor da água e para a concentração de cobre classificaram o arroio Demétrio como Classe 4 em geral, ou seja, esta água é adequada apenas para navegação e harmonia da paisagem. As principais alterações genotóxicas (micronúcleos e brotos nucleares) foram observadas no P2, no qual foram obtidos os maiores teores de cobre, além de ferro e manganês. Intervenções antrópicas foram observadas em P1 e P2; no entanto, devido à sua baixa concentração de metais, o P3, próximo ao rio Gravataí, manifestou uma melhoria na qualidade ambiental.

Water quality assessment of the Demetrio stream: an affluent of the Gravataí River in the South of Brazil / Qualidade da água do arroio Demetrio: um afluente do rio Gravataí no sul do Brasil

The Gravataí River basin, one of the main water sources of the metropolitan region of Porto Alegre, is among the ten most polluted rivers in Brazil. Water quality is monitored only through physico-chemical and microbiological parameters in Brazil, and in this context, considering the importance of the use of biomarkers in complementing the analysis of water, the present study aimed to evaluate the environmental quality of the main affluent of the Gravataí River, Demetrio stream, through physico-chemical, microbiological, and cytogenotoxic criteria, at the stream source (P1), whereas samples P2 and P3 were obtained from the upstream near the area with the highest urban density and the downstream near the meeting point with the Gravataí River, respectively. The results for copper concentration and color classified the Demetrio stream as Class 4 in general, that is, the water is suitable only for navigation and to landscape harmony. The main genotoxic alterations (micronuclei and nuclear buds) were observed in P2, in which were obtained the highest levels of copper, in addition to iron and manganese. Anthropic interventions were observed in P1 and P2; however, due to its low metal concentration, P3, near the Gravataí River, manifested an improvement in environmental quality.

A bacia do rio Gravataí, uma das principais fontes de água da região metropolitana de Porto Alegre, está entre os dez rios mais poluídos do Brasil. No Brasil a qualidade da água é monitorada apenas através de parâmetros físico-químicos e microbiológicos e, nesse contexto, considerando a importância do uso de biomarcadores para complementar a análise da água, o presente estudo teve como objetivo avaliar a qualidade ambiental do principal afluente do Rio Gravataí, o arroio Demétrio, através de critérios físico-químicos, microbiológicos e citogenotóxicos, na nascente do arroio (P1), a montante e próximo à área com maior densidade urbana (P2) e a jusante e próximo ao ponto de encontro com o rio Gravataí (P3). Os resultados para a cor da água e para a concentração de cobre classificaram o arroio Demétrio como Classe 4 em geral, ou seja, esta água é adequada apenas para navegação e harmonia da paisagem. As principais alterações genotóxicas (micronúcleos e brotos nucleares) foram observadas no P2, no qual foram obtidos os maiores teores de cobre, além de ferro e manganês. Intervenções antrópicas foram observadas em P1 e P2; no entanto, devido à sua baixa concentração de metais, o P3, próximo ao rio Gravataí, manifestou uma melhoria na qualidade ambiental.

Water quality assessment of the Demetrio stream: an affluent of the Gravataí River in the South of Brazil

Abstract The Gravataí River basin, one of the main water sources of the metropolitan region of Porto Alegre, is among the ten most polluted rivers in Brazil. Water quality is monitored only through physico-chemical and microbiological parameters in Brazil, and in this context, considering the importance of the use of biomarkers in complementing the analysis of water, the present study aimed to evaluate the environmental quality of the main affluent of the Gravataí River, Demetrio stream, through physico-chemical, microbiological, and cytogenotoxic criteria, at the stream source (P1), whereas samples P2 and P3 were obtained from the upstream near the area with the highest urban density and the downstream near the meeting point with the Gravataí River, respectively. The results for copper concentration and color classified the Demetrio stream as Class 4 in general, that is, the water is suitable only for navigation and to landscape harmony. The main genotoxic alterations (micronuclei and nuclear buds) were observed in P2, in which were obtained the highest levels of copper, in addition to iron and manganese. Anthropic interventions were observed in P1 and P2; however, due to its low metal concentration, P3, near the Gravataí River, manifested an improvement in environmental quality.

Resumo A bacia do rio Gravataí, uma das principais fontes de água da região metropolitana de Porto Alegre, está entre os dez rios mais poluídos do Brasil. No Brasil a qualidade da água é monitorada apenas através de parâmetros físico-químicos e microbiológicos e, nesse contexto, considerando a importância do uso de biomarcadores para complementar a análise da água, o presente estudo teve como objetivo avaliar a qualidade ambiental do principal afluente do Rio Gravataí, o arroio Demétrio, através de critérios físico-químicos, microbiológicos e citogenotóxicos, na nascente do arroio (P1), a montante e próximo à área com maior densidade urbana (P2) e a jusante e próximo ao ponto de encontro com o rio Gravataí (P3). Os resultados para a cor da água e para a concentração de cobre classificaram o arroio Demétrio como Classe 4 em geral, ou seja, esta água é adequada apenas para navegação e harmonia da paisagem. As principais alterações genotóxicas (micronúcleos e brotos nucleares) foram observadas no P2, no qual foram obtidos os maiores teores de cobre, além de ferro e manganês. Intervenções antrópicas foram observadas em P1 e P2; no entanto, devido à sua baixa concentração de metais, o P3, próximo ao rio Gravataí, manifestou uma melhoria na qualidade ambiental.

Water quality assessment of the Demetrio stream: an affluent of the Gravataí River in the South of Brazil.

The Gravataí River basin, one of the main water sources of the metropolitan region of Porto Alegre, is among the ten most polluted rivers in Brazil. Water quality is monitored only through physico-chemical and microbiological parameters in Brazil, and in this context, considering the importance of the use of biomarkers in complementing the analysis of water, the present study aimed to evaluate the environmental quality of the main affluent of the Gravataí River, Demetrio stream, through physico-chemical, microbiological, and cytogenotoxic criteria, at the stream source (P1), whereas samples P2 and P3 were obtained from the upstream near the area with the highest urban density and the downstream near the meeting point with the Gravataí River, respectively. The results for copper concentration and color classified the Demetrio stream as Class 4 in general, that is, the water is suitable only for navigation and to landscape harmony. The main genotoxic alterations (micronuclei and nuclear buds) were observed in P2, in which were obtained the highest levels of copper, in addition to iron and manganese. Anthropic interventions were observed in P1 and P2; however, due to its low metal concentration, P3, near the Gravataí River, manifested an improvement in environmental quality.

Correlation of plasma butyrylcholinesterase concentration with Acethylcholinesterase H353N polymorphism in the inflammatory response of Chagas disease patients.

Trypanosoma cruzi infection triggers an intense production of pro-inflammatory cytokines mediated by T helper 1 lymphocytes, inducing the anti-inflammatory reflex of acetylcholine (ACh). The ACh concentration modulation is associated to the two major esterases, the acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). AChE H353N protein polymorphism is related to low Chagas chronic disease prognostic. In order to evaluate the correlation of plasmatic BuChE concentration and the presence of AChE H353N polymorphism in Chagas disease patients and healthy individuals, we studied two groups of individuals, one of 61 Chagas disease patients and another of 74 healthy individuals. Plasma concentration of BuChE was measured by the chemiluminescent method and AChE H353N polymorphism was investigated by PCR-RFLP and sequencing of the respective encoding AChE gene fragment. The BuChE concentration was statistically higher in Chagas disease patients, with no AChE genotype significant influence. AChE genotypes YT*A/YT*A, YT*A/YT*B and YT*B/YT*B, respectively, were expressed in 53 (86.88%), 7 (11.46%) and one (1.64%) chagasic patients, and in 68 (91.89%), 6 (8.10%) and none healthy individuals. BuChE activity may represent an important marker for chronic Chagas disease inflammatory process and prognostic. Lower BuChE concentration correlated with AChE YT*B allele, although without statistical power.

Paradoxical effects of vitamin C in Chagas disease.

Trypanosoma cruzi infection stimulates inflammatory mediators which cause oxidative stress, and the use of antioxidants can minimize the sequelae of Chagas disease. In order to evaluate the efficacy of vitamin C in minimizing oxidative damage in Chagas disease, we orally administered ascorbic acid to Swiss mice infected with 5.0 × 104 trypomastigote forms of T. cruzi QM2 strain. These animals were treated for 60 days to investigate the acute phase and 180 days for the chronic phase. During the acute phase, the animals in the infected and treated groups demonstrated lower parasitemia and inflammatory processes were seen in more mice in these groups, probably due to the higher concentration of nitric oxide, which led to the formation of peroxynitrite. The decrease in reduced glutathione concentration in this group showed a circulating oxidant state, and this antioxidant was used to regenerate vitamin C. During the chronic phase, the animals in the infected and treated group showed a decrease in ferric reducing ability of plasma and uric acid concentrations as well as mobilization of bilirubin (which had higher plasma concentration), demonstrating cooperation between endogenous non-enzymatic antioxidants to combat increased oxidative stress. However, lower ferrous oxidation in xylenol orange concentrations was found in the infected and treated group, suggesting that vitamin C provided biological protection by clearing the peroxynitrite, attenuating the chronic inflammatory process in the tissues and favoring greater survival in these animals. Complex interactions were observed between the antioxidant systems of the host and parasite, with paradoxical actions of vitamin C.

In vitro lingual bracket evaluation of indirect bonding with plasma arc, LED and halogen light.

OBJECTIVES: Evaluate the shear bond strength (SBS) and the adhesive remnant index (ARI) of indirect bonded lingual brackets using xenon plasma arc light, light-emitting diode (LED) and conventional quartz-tungsten-halogen light. MATERIAL AND METHODS: Lingual brackets were bonded indirectly to 60 premolars divided to three groups according to the curing light used: Group 1, plasma arc for 6 s; Group 2, LED for 10 s; and Group 3, halogen light for 40 s. After bonding, the specimens were subjected to a shear force until debonding. The debonding pattern was assessed and classified according to the ARI scores. The mean shear bond strengths were accessed by anova followed by the Student-Newman-Keuls test for multiple comparisons. ARI scores were assessed using the chi-square test. RESULTS: The three groups showed significant differences (p < 0.001), with the averages of group 1 < group 2 < group 3. Groups showed no differences regarding ARI scores. CONCLUSION: Bonding lingual brackets indirectly with plasma arc, during 60% of the time used for the LED, produced lower SBS than obtained with the latter. Using LED during 25% of the time of the halogen light produced lower SBS than obtained with the latter. These differences did not influence the debonding pattern and are clinically acceptable according to the literature.

[Susceptibility of Rhodnius neglectus, Rhodnius robustus and Triatoma infestans (Hemiptera, Reduviidae, Triatominae) to infection by two strains of Trypanosoma cruzi (Kinetoplastidae, Trypanosomatidae) using artificial xenodiagnosis]. / Susceptibilidade de Rhodnius neglectus, Rhodnius robustus et Triatoma infestans (Hemiptera, Reduviidae, Triatominae) à infecçao por duas cepas de Trypanosoma cruzi (Kinetoplastidae, Trypanosomatidae) utilizando xenodiagnóstico artificial.

The susceptibility of 3rd instar nymph of Triatominae Rhodnius neglectus, R. robustus and Triatoma infestans to Trypanosoma cruzi Y and AMJM strains was verified using artificial xenodiagnosis. After the accomplishment of the xenodiagnosis, the faeces of the Triatominae were analyzed on two-day intervals from day 5 until day 31 post infection, using the abdominal compression technique. The results showed differences in the susceptibility of the Triatominae for the two strains studied, and the optimal period reading differed from day 11 to day 19 for the Y strain and from day 11 to day 15 for the AMJM strain. For the Y strain, all three Triatominae species showed good susceptibility, whereas in the AMJM strain, the highest susceptibility was observed with R. neglectus, followed by T. infestans and R. robustus.

DNA vaccination against persistent viral infection.

This study shows that DNA vaccination can confer protection against a persistent viral infection by priming CD8+ cytotoxic T lymphocytes (CTL). Adult BALB/c (H-2d) mice were injected intramuscularly with a plasmid expressing the nucleoprotein (NP) gene of lymphocytic choriomeningitis virus (LCMV) under the control of the cytomegalovirus promoter. The LCMV NP contains the immunodominant CTL epitope (amino acids 118 to 126) recognized by mice of the H-2d haplotype. After three injections with 200 micrograms of NP DNA, the vaccinated mice were challenged with LCMV variants (clones 13 and 28b) that establish persistent infection in naive adult mice. Fifty percent of the DNA-vaccinated mice were protected, as evidenced by decreased levels of infectious virus in the blood and tissues, eventual clearance of viral antigen from all organs tested, the presence of an enhanced LCMV-specific CD8+ CTL response, and maintenance of memory CTL after clearance of virus infection. However, it should be noted that protection was seen in only half of the vaccinated mice, and we were unable to directly measure virus-specific immune responses in any of the DNA-vaccinated mice prior to LCMV challenge. Thus, at least in the system that we have used, gene immunization was a suboptimal method of inducing protective immunity and was several orders of magnitude less efficient than vaccination with live virus. In conclusion, our results show that DNA immunization works against a persistent viral infection but that efforts should be directed towards improving this novel method of vaccination.

Independent fluctuation of human immunodeficiency virus type 1 rev and gp41 quasispecies in vivo.

The human immunodeficiency virus type 1 (HIV-1) overlapping rev and env coding sequences have been examined from sequential peripheral blood mononuclear cell DNA samples from one individual. These were the same DNA samples from which sequence data for the tat and nef/long terminal repeat loci have been derived and span a 4-year period. The rev/env sequences were established by sequencing cloned polymerase chain reaction products. The structure of the populations of rev protein sequences increased in complexity with disease, while those of the corresponding env sequences remained complex. This suggests that the rev and env populations evolved differently, probably reflecting different selection pressures. No defective rev variants encoded substitutions in residues 76 through 79, indicating that the experimental finding of down regulation of rev activity by competitive inhibition may not necessarily occur in vivo. After having analyzed three HIV loci (15% of the genome) from the same individual over 4 years, it is clear that no two loci evolved similarly, indicating the difficulties in comparing data from different loci.

LAV revisited: origins of the early HIV-1 isolates from Institut Pasteur.

Two of the first human immunodeficiency virus type-1 (HIV-1) strains isolated were authenticated by reanalyzing original cultured samples stored at the Collection Nationale de Culture des Microorganismes as well as uncultured primary material. Cloned polymerase chain reaction products were used to analyze coding sequences of the V3 loop in the gp120 glycoprotein. The original isolate HIV-1 Bru, formerly called LAV, was derived from patient BRU. HIV-1 Lai was derived from patient LAI and contaminated a HIV-1 Bru culture between 20 July and 3 August 1983. The culture became, in effect, HIV-1 Lai, identifiable by a unique motif in the V3 loop. Because of this contamination two, rather than one, HIV-1 isolates were sent to the Laboratory of Tumor Cell Biology at the National Cancer Institute on 23 September 1983. Original HIV-1 Bru was indeed present in the sample marked JBB/LAV. However the M2T-/B sample harbored HIV-1 Lai, a strain capable of growing on established cell lines. The striking similarity between HIV-1 Lai (formerly LAV-Bru) and HTLV-3B sequences remains.

AIDS virus-specific cytotoxic T lymphocytes in lung disorders.

Human immunodeficiency virus (HIV) is implicated in the development of AIDS (acquired immune deficiency syndrome). HIV infection leads to the generation of HIV-specific thymus-derived (T) lymphocytes in humans and apes. We describe an experimental system permitting the quantitative and systematic analysis of HIV-specific cytotoxic T lymphocytes (CTL). Functional, HIV-specific CTL are obtained by broncho-alveolar lavage (BAL) from the lungs of seropositive patients with lymphocytic alveolitis. These alveolar CTL: (1) recognize and kill HIV-infected alveolar macrophages in vitro under autologous, but not heterologous, conditions; (2) correspond to standard CTL as they express the CD3 and CD8 surface markers, but not the CD4 marker; and (3) are restricted by class I HLA transplantation antigens in their cytotoxic activities. We propose the hypothesis that interactions between HIV-specific CTL and infected macrophages induce major inflammatory reactions in seropositive patients.

Avaliação da relação ântero-posterior dos maxilares: em descendentes de Japoneses Ortognatas e Prognatas / Analysis of maxillary anterior-posterior relation in prognate and orthognate japanese descendents

Os autores estudaram uma amostra de 50 indivíduos descendentes de japoneses residentes em Araraquara/SP; destes, 25 apresentavam oclusão aceitável e perfil harmônico e 25 apresentavam oclusões dentárias caracterizadas com más-oclusões de Classe III (ANGLE) e perfil côncavo. Telerradiografias destes indivíduos foram analisadas, com o objetivo de determinar as relações ântero-posteriores dos maxilares, segundo os critérios preconizados por RIEDEL (13) (ângulo ANB). JACOBSON (11) (distância AO-BO); DOWNS (5) (ângulo NPg.AB) e SCHWARZ (16) (PP.AB). Os resultados mostraram que estes indivíduos, divididos em grupos de prognatas e ortognatas, distinguem-se não só quanto ao conjunto dos critérios citados, como também, segundo cada um deles. Cada critério analisado foi, por si só, suficiente para discriminar as displasias ântero-posteriores dos maxilares. Os diferentes critérios utilizados mostraram correlação estatisticamente significante para o grupo dos prognatas, enquanto que para o grupo dos ortognatas não houve correlação estatisticamente significante com excessão de ANB com NPg.AB