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Dules auriga, a native Brazilian teleost, was applied as a sentinel species regarding metal contamination at Ilha Grande Bay, previously considered a reference site in Southeastern Brazil. Cytosolic (S50) and metallothionein-bound (HTS50) hepatic iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), cadmium (Cd), and silver (Ag) were determined by inductively coupled plasma optical emission spectrometry (ICP-OES), while metallothionein (MT) concentrations were determined by polarography. Ag concentrations in both cytosolic fractions were below the limit of detection. All other HTS50 metal contents were significantly lower than S50 contents. No significant associations were found for MT. Fe and Mn S50 were positively and moderately correlated to total length, as well as HTS50 Mn, while total weight was correlated to both Mn fractions, suggesting that environmental Mn and Fe concentrations may influence fish growth. A moderate correlation between the condition factor and the S50 Cu fraction was observed, also indicating that Cu may affect fish growth. Inter-element correlations were observed, including between Cd, a toxic element, and Mn and Zn, both essential elements. Calculated molar ratios indicate that both Mn and Zn are in molar excesses compared with Cd, corroborating literature assessments regarding protective Mn and Zn effects against Cd. Lack of MT correlations suggests that metal concentrations may not be high enough to reach an MT induction threshold and that MT variability is probably linked to environmental metal concentrations. Therefore, the increased environmental contaminant levels observed in the study area indicate the need for biomonitoring efforts aiming at the application of efficient mitigation measures.

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The identification of estuarine sentinel species is of paramount importance. The potential of the species Dules auriga sampled from Guanabara Bay, Brazil regarding metal contamination was assessed. Hepatic metallothionein (MT) and Fe, Zn, Cu, Mn, Cd and Ag concentrations were determined in cytosolic fractions (S50 and HT S50) by polarography and ICP-OES, respectively. HT S50 Fe, Mn and Zn were lower than in the S50 fraction, indicating MT-detoxification. MT was correlated to HT S50 Zn, indicating Zn homeostasis. Zn was negatively correlated to weight (TW) and length (TL), suggesting environmental Zn influence. A moderate negative correlation between HT S50 Cu and the condition factor (CF) was observed, indicating that increasing environmental Cu concentrations may decrease D. auriga CF. Several inter-elemental correlations were observed. No MT, TW and TL correlations were found. Thus, MT variability is probably linked to environmental metal concentrations. D. auriga fulfills most sentinel requisites, and MT may be an adequate metal contamination biomarker in this species.

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Metal speciation analysis in MTs was carried out in two tropical fish species of Brazil, the freshwater fish pearl cichlid (Geophagus brasiliensis) and the marine fish white sea catfish (Netuma barba), that are presently used to monitor the effects of heavy metal pollution in aquatic ecosystems in Brazil. In order to obtain the MT fraction, liver cytosols from both fish species where subjected to size exclusion fractionation, monitoring on-line the metal signal (Cd, Cu and Zn) by ICP-MS while protein elution was followed by on-line UV detection. That MT fraction was then separated by anion-exchange (AE)-FPLC, whose optimal chromatographic conditions were optimized for the separation of the different hepatic MT isoforms existing in both fish species. Specific detection of separated metalloforms was carried out again by the hyphenation of the AE chromatographic system with the ICP-MS instrument. The analytical results showed that MTs of these fish species, unknown so far, exhibited unique characteristics in comparison with standard MTs and other fish liver MTs. In fact, MT isoforms of N. barba turned out to be very anionic, as indicated by their high retention in the Mono Q column and the strong ionic strength required to separate them. As for G. brasiliensis, cadmium was exclusively present in only one of the peaks of the MT isoforms showing a unique metal-binding behavior for MT in this fish species. The differences between the MTs among these species and the different association of metals in particular MT isoforms display the importance of the metal speciation analysis in these proteins prior to its use as bioindicators.

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The use of isotope dilution analysis (IDA) with inductively coupled plasma-mass spectrometry (ICP-MS) for the determination of oxidized metallothionein (MT) by a Cd-saturation method is investigated. The method developed here is a modification of an earlier methodology which used a radioactive Cd isotope ((109)Cd). While retaining the many advantages of this previous approach, the procedure presented here uses stable isotope ratio measurements ((114)Cd/(111)Cd) for the determination of MT. Experimental parameters governing the instrumental precision and accuracy for isotope ratio measurements of Cd by ICP-MS were characterized. Systematic errors, including mass bias, detector dead time, and spectroscopic interferences, could be easily corrected. The isotope dilution ICP-MS method was validated by the determination of very low levels of cadmium in biological certified reference materials (NIST SRM 2670 freeze-dried urine, IAEA H-8 horse kidney, and BCR TP-25 lichens). Finally, the IDA procedure was evaluated for the determination of oxidized MT by a Cd-saturation method previously developed using radioactive (109)Cd. The final procedure was applied to the quantification of MT in Long-Evans Cinnamon rat liver cytosol samples and the results were compared with data obtained for the same samples using the reference (109)Cd methodology. A good agreement between the analytical values obtained by both methods was observed.

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Chagas disease is becoming a public health problem in Latin America due to the wide distribution, the high prevalence, the magnitude of the damage caused and the difficulties to control it. In Paraguay, the disease is mainly distributed in the departments of Paraguarí, Cordillera and Central. Prevalence in marginal zones, where migrations from rural populations and endemic areas make possible the urbanization of the disease, has no been studied yet. This is a descriptive study with a cross-sectional sampling and a probabilistic system recruitment carried out in school aged children from marginal zones of Asunción to determine the prevalence of Chagas' disease. Serological methods, parasite isolation and questionnaires were used to achieve the goals. Nine hundred and fifty three children were studied to determine the prevalence of Chagas' disease in marginal zones which was 1.4%.

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In order to evaluate the significance of alpha- and beta-fragments of metallothionein with regard to Cd binding in biosynthetic processes, the Cd-binding ability of four mutant metallothioneins was examined using the Escherichia coli expression system. The features of the mutant metallothioneins were proteins in which cysteine residues in the alpha- or beta-fragment were replaced with alanine residues, or that the sequential order of the fragments was altered. The expressed mutant metallothioneins having an intact alpha-fragment showed the constructive abilities of the Cd-thiolate cluster. On the other hand, mutant metallothionein having an alpha-fragment lacking metal-binding sites exhibited no Cd-binding ability. The condition for maintaining the Cd-binding capability of metallothionein was that the alpha-fragment remains intact irrespective of the sequential order of the two fragments. The alpha-fragment is an indispensable component in metal-binding processes of Cd-metallothionein.

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Five bisbenzylisoquinoline (BBI) alkaloids, curine, cycleanine, isotet:andrine, limacine and pheanthine were tested for trypanocidal activity in C 3H He mice infected with Y or CL strain of Trypanosoma cruzi. The activity was compared with the baseline drug, benznidazole. Oral treatment was more effective with curine at 10 mg/kg or with cycleanine at 2 mg/kg daily for 10 days in mice infected with Y or CL strain. In these groups, the parasitemias were negative after 5-7 weeks after inoculation and mortality time 50 (MT(50)) was significantly higher than untreated mice. Benznidazole was effective in mice infected with CL strain but not in mice infected with Y strain. The other BBI showed a relative efficacy against both strains. The effect of BBI alkaloids could be due to a blocking of the Ca2+ channel for the regulation of T. cruzi infectivity to invade host cells or their selective immunosuppressive properties.

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We examined the independent self-assembly of the alpha- and beta-fragments of human metallothionein (MT) into cadmium-binding conformation in an Escherichia coli expression system, in addition to wild-type MT expression. The expressed alpha-fragment formed independently the structure of a metal-binding cluster without the aid of the beta-fragment. The alpha-fragment and wild-type MT expressed in E.coli were purified and analyzed for their biochemical and spectroscopic properties. The apparent cadmium binding of the alpha-fragment was approximately 12-fold greater than that for the wild-type MT, whereas in other respects the studied biochemical properties were similar. In contrast, we were unable to obtain any independently expressed beta-fragment as the cadmium-binding form in this study. Possible explanations for this phenomenon are discussed.

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A genetic approach was undertaken to investigate the physiological roles of human metallothionein-2. A constructed expression plasmid, pEXPMTII, in which human metallothionein-IIA cDNA was inserted downstream of a tryptophan-lactose promoter, was used to transform Escherichia coli JM105 strain. Cadmium-binding metallothionein was successfully expressed in E. coli in the medium containing cadmium, while copper and zinc-metallothioneins were scarcely observed in copper- or zinc-containing medium. The amino acid composition and sequence of the biosynthesized cadmium-metallothionein were analyzed. The selectivity of metals bound to metallothionein and the stability of metal-binding forms of metallothionein in E. coli were discussed. In addition, cadmium, zinc, or copper resistance of the cells expressing metallothionein was examined. Cells transformed with the plasmid pEXPMTII and cultured in a medium containing cadmium exhibited tolerance only to cadmium. It was demonstrated that human metallothionein-2 functioned for cadmium detoxification in E. coli.

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Insecticidal residual effect and triatomine infestation rates in houses of a community fumigated with lambdcyhalothrin (Icon) are reported. No mortality was observed in 5th-instar Triatoma infestans nymphs in 72-hr exposure test on three different surfaces, one month after fumigation for a dose of 31.5 mg am/m2. However, during post-exposure observation a mortality of 60% was recorded for those insect exposed on sprayed woodboard. The results observed with mud-containing treated walls, were markedly poorer (0% of mortality). Twelve month after spraying 40% of mortality was observed on first-instar T. infestans nymphs in 72-hr exposure test on woodboard, but lower mortality rates were observed in mud-containing materials. When the effect of deltamethrin (109 mg ai/m2) and lambdcyhalothrin (94 mg ai/m2) was compared, the former did not appear to be superior at similar loads. Both have showed a mortality rate of 30% on 5th-instar T. infestans nymphs three months post-fumigation. The dose utilized in the field fumigation was enough to get a significant (p < 0.0001) control of triatomine domestic infestation, since it was sufficient to keep 95% of the houses uninfested throughout 21 months following treatment, when compared with baseline situation. A remarkable knock-down effect on adult and nymphs forms of the insect and a high in situ mortality were observed as a result of its application, even at very low doses.

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Animal reservoirs for Trypanosoma cruzi infection were investigated in 5 communities in the Department of San Pedro, currently one of Paraguay's most highly endemic areas. A total of 112 domestic animals (37 cattle, 2 horses, 1 ass, 20 pigs, 44 dogs and 8 cats) and 4 wild animals (1 white-eared opossum, 2 yellow armadillos and 1 common long-nosed armadillo) were examined for blood. Although no trypomastigotes were found by 2 direct observation methods, the microhaematocrit and Giemsa stained thick and thin smears methods, several forms of trypanosoma flagellates morphologically identical to T. cruzi were detected in the liver infusion tryptose (LIT) medium from a single sample taken from a yellow armadillo, Euphractus sexicintus. When serum samples of all the animals were examined for antibody to T. cruzi by direct agglutination (DA) test, 3 cattle, 2 pigs, 16 dogs and 3 cats had positive titers (1:32 or greater), but no wild animals showed positive reactions. T. cruzi was not found by culture nor microscopic examination of samples from any of the seropositive animals. However, domestic animals such as cattle, pigs, dogs and cats which were found to be seropositive in this study, possibly act as an animal reservoir in this endemic area as well as armadillos in which T. cruzi was observed.

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Eleven Paraguayan strains of Trypanosoma cruzi, from Chagas' disease patients and the bug vectors, were examined by restriction endonuclease analysis of kinetoplast DNA using Hae III, Msp I, Eco RI, HinfI, Taq I and Rsa I. Four schizodeme-profile groups were identified. Group 1 had much simpler profiles than groups 2, 3 and 4 and, although there were homogeneous profiles in the latter three groups, each group could be distinguished from the others. The profiles of group 1 could not be matched with any of the standard strains from Brazil, Chile and Columbia included in the schizodeme comparison. The profiles of groups 3 and 4 shared most features with those standards of the Brazilian Z2 zymodeme.

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The authors discuss the production and use of Brazilian labor market data. Noting improvements in data collection during the 1980s, "this study aims to point out the common points, the differences and the problems of the various data sources [that call] for urgent revisions. It concludes that any attempt in this way will necessarily lead to a more intense and democratic process of consultation among the producers and...the users...." (SUMMARY IN ENG)

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A list of fifty-two plant species used for medicinal purposes or to control arthropods by peasants near Belo Horizonte, Minas Gerais (Brazil) has been compiled. Preparation procedures are also noted. The use of plants for controlling insects and to heal a condition known as "ferida brava" is discussed. The role of indigenous plant remedies within the local health care system is worthy of a closer investigation.

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Twenty four Cebus apella monkeys were studied as a biological model for the cardiac chronic form of Chagas' disease. Twelve were inoculated with Trypanosoma cruzi trypomastigotes, seven with the Brazilian Y strain and five with the Argentinian RA strain. Twelve monkeys were uninfected controls. The following parameters were studied: body weight, body temperature, direct parasitemia, xenodiagnosis, specific antibodies by IFA, clinical chemistry, hematology, ECG and chest X-ray. Three monkeys infected with Y strain were sacrificed at 4 months and 4 monkeys at 12 months after inoculation. Monkeys inoculated with RA strain were sacrificed at 48 months. Direct parasitemia was positive within a week after inoculation in all monkeys. Xenodiagnosis was positive until 49.0 +/- 3.0 and 79.0 +/- 6.0 weeks p.i. for Y and RA strains, respectively. In all inoculated monkeys an increase in antibody titers was detected within 3 weeks after inoculation. In all monkeys inoculated with the Y strain and 3/5 with the RA strain abnormal ECGs were observed within 1 or 2 weeks p.i., becoming more severe in the chronic phase. Y strain inoculated monkeys sacrificed at 4 months presented only a slight concentric hypertrophy of the heart left ventricle. Those sacrificed at 12 months had concentric left ventricle hypertrophy and 3/4 had an aneurism of the apex. Four out of 5 RA strain inoculated monkeys had an enlarged, flaccid heart; 3/5 aneurism of the apex and 2/5 concentric hypertrophy of the left ventricle.(ABSTRACT TRUNCATED AT 250 WORDS)

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PIP: This is the second part of a study evaluating Brazil's demographic and agricultural censuses and their divergent representations of change in agricultural employment. Consideration is given to occupational status, economic activity, regional variations, and residence characteristics of agricultural workers. The effect of censal differences on the measurement of seasonal fluctuations in employment and the labor force participation of women and children is noted. (SUMMARY IN ENG)^ieng

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Eugenia uniflora is widely used in Paraguayan folk medicine. A hydroalcoholic extract of the leaves showed some central nervous system activity in hippocratic screening when given intraperitoneally, but little to no acute or subacute toxicity in doses up to 4200 mg/kg orally in BALB c mice. The LD50 of the extract was 220 mg/kg i.p. in mice. A decoction or infusion of the leaves is recommended for treating gout by native herbalists. The known flavonoids quercitrin, quercetin, myricitrin and myricetin were found to be responsible for the xanthine oxidase inhibitory action of the plant extract.

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Data collected from population and agricultural censuses concerning agricultural employment in Brazil are compared. "This study aims at analyzing the origin, the form and the significance of the differences between the two censuses. In this first part a detailed comparison is made between their respective concepts, their methodologies, their limitations and their outcomes. The comparison goes back even to the first modern censuses in order to evaluate the evolution of censal instruments and its importance for the...understanding of employment in agriculture." Other data sources are also considered. (SUMMARY IN ENG)

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