Urban malaria in the Brazilian Western Amazon Region I: high prevalence of asymptomatic carriers in an urban riverside district is associated with a high level of clinical malaria

Cross sectional studies on malaria prevalence was performed in 2001, 2002, and 2004 in Vila Candelária, an urban riverside area of Porto Velho, Rondônia, in the Brazilian Western Amazon, followed by longitudinal surveys on malaria incidence. Vila Candelária is a working class district, provided with electricity, water supply, and basic sanitation. Previous preliminary surveys indicated high malaria incidence in this community. At the end of year 2000 regular diagnostic and treatment measures for malaria were introduced, with active search of febrile cases among residents. Despite of both rapid treatment of cases and relative good sanitary and housing conditions, the malaria incidence persisted at high levels during the following years with an annual parasite index of 150 to 300/1000 inhabitants. Parasite surveys in 2001, 2002, and 2004 achieved through microscopy and polymerase chain reaction to diagnose malaria showed a constant high prevalence of asymptomatic carriers for both Plasmodium falciparum and P. vivax parasites. It was concluded that asymptomatic carriers represent an important reservoirs of parasites and that the carriers might contribute to maintaining the high level of transmission. Comparing our findings to similar geo-demographic situations found in other important urban communities of the Brazilian Amazon, we propose that asymptomatic carriers could explain malaria's outbreaks like the one recently observed in Manaus.

Prevalence and level of antibodies to the circumsporozoite protein of human malaria parasites in five states of the Amazon region of Brazil

The aim of this study was to determine the prevalence of malaria infection and antibodies against the repetitive epitopes of the circumsporozoite (CS) proteins of Plasmodium falciparum, P. malariae, P. vivax VK210, P. vivax VK247, and P. vivax-like in individuals living in the states of Rondônia, Pará, Mato Grosso, Amazonas, and Acre. Active malaria transmission was occurring in all studied sites, except in Acre. P. falciparum was the predominant species in Pará and Rondônia and P. vivax in Mato Grosso. Infection by P. malariae was low but this Plasmodium species was detected in Rondônia (3.5%), Mato Grosso (2.5%), and Pará (0.8%). High prevalence and levels of serological reactivity against the CS repeat peptides of P. falciparum were detected in Rondônia (93%) and Pará (85%). Sera containing antibodies against the CS repeat of P. malariae occurred more frequently in Rondônia (79%), Pará (76%), and Amazonas (68%). Antibodies against the repeat epitope of the standard CS protein of P. vivax VK210, P. vivax VK247, and P. vivax-like were more frequent in Rondônia, Pará, and Mato Grosso. The high frequency of reactions to P. malariae in most of the areas suggests that the infection by this Plasmodium species has been underestimated in Brazil.

The level of ascorbate peroxidase is enhanced in benznidazole-resistant populations of Trypanosoma cruzi and its expression is modulated by stress generated by hydrogen peroxide

Ascorbate peroxidases (APX) are class I heme-containing enzymes that convert hydrogen peroxide into water molecules. The gene encoding APX has been characterized in 11 strains of Trypanosoma cruzi that are sensitive or resistant to benznidazole (BZ). Bioinformatic analysis revealed the presence of two complete copies of the T. cruzi APX (TcAPX) gene in the genome of the parasite, while karyotype analysis showed that the gene was present in the 2.000-kb chromosome of all of the strains analyzed. The sequence of TcAPX exhibited greater levels of similarity to those of orthologous enzymes from Leishmania spp than to APXs from the higher plant Arabidopsis thaliana. Northern blot and real-time reverse transcriptase polymerase chain reaction (RT-PCR) analyses revealed no significant differences in TcAPX mRNA levels between the T. cruzi strains analyzed. On the other hand, Western blots showed that the expression levels of TcAPX protein were, respectively, two and three-fold higher in T. cruzi populations with in vitro induced (17 LER) and in vivo selected (BZR) resistance to BZ, in comparison with their corresponding susceptible counterparts. Moreover, the two BZ-resistant populations exhibited higher tolerances to exogenous hydrogen peroxide than their susceptible counterparts and showed TcAPX levels that increased in a dose-dependent manner following exposure to 100 and 200 µM hydrogen peroxide.

Correlation between the infestation level of Sitophilus zeamais Motschulsky, 1855 (Coleoptera, Curculionidae) and the quality factors of stored corn, Zea mays L. (Poaceae)

This experiment was carried out in order to evaluate the effect of Sitophilus zeamais on physical, physiological and sanitary quality of stored corn. Samples of 500 g of the hybrid OC-705, in three replicates, were conditioned in glasses covered with a screened lid, and kept in chamber at 25±2ºC, 70±5% RH and 12 h of photophase, for 150 days. The infestation levels were 0, 5, 15 and 50 adults/replicate, for the storage periods of 30, 60, 90, 120 and 150 days. The moisture content, classification, weight loss, germination and internal infestation were evaluated monthly. Significant inverse correlations were verified between the number of insects and both the germination and the weight loss; also between the internal infestation and the germination and the standard type. The presence of S. zeamais showed a positive correlation with the weight loss, what means that the internal and external infestations contribute to the reduction of physiological and physical quality of corn seeds. The mean dry matter loss was 0,36%/day, corresponding to a consumption of 0,0001%/insect/month. As the result of those damages, the product suffered reduction of the commercial grade in 30 days, with significant loss in all quality factors.

Rice yield estimation based on weather conditions and on technological level of production systems in Brazil

The objective of this work was to evaluate an estimation system for rice yield in Brazil, based on simple agrometeorological models and on the technological level of production systems. This estimation system incorporates the conceptual basis proposed by Doorenbos & Kassam for potential and attainable yields with empirical adjusts for maximum yield and crop sensitivity to water deficit, considering five categories of rice yield. Rice yield was estimated from 2000/2001 to 2007/2008, and compared to IBGE yield data. Regression analyses between model estimates and data from IBGE surveys resulted in significant coefficients of determination, with less dispersion in the South than in the North and Northeast regions of the country. Index of model efficiency (E1') ranged from 0.01 in the lower yield classes to 0.45 in higher ones, and mean absolute error ranged from 58 to 250 kg ha‑1, respectively.

Relation between the level of self-mutilation and theconcentration of fecal metabolites of glucocorticoids incaptive chimpanzees (Pan troglodytes)

The influence of stress in an environment, according with the behavioral and endocrine variables of primates, are increasingly being studied by a diversity of authors, and have shown that abnormal behaviors associated with increased glucocorticoids may be directly related with the impairment of their well-being. In this work were used 22 adult chimpanzees (Pan troglodytes), 11 males and 11 females, kept in captivity in three different institutions. All animals had their behavior registered by focal session using a 30 seconds sample interval, during six months, totaling 4,800 registries per each animal. During this period, fecal samples were collected 3 times a week for the extraction and measurement of the concentration of fecal metabolites of glucocorticoid by radioimmunoassay. Of the total observed, stereotypical behaviors represented 13,45±2.76%, and among them, self-mutilation represented 38.28±3.98 %. The animals were classified into three different scores, according with the percentage of body surface with alopecia due to self-mutilation. It was found a positive correlation of high intensity between the scores of alopecia due to the observed mutilation and the average concentrations of fecal metabolites of glucocorticoids. This result strongly suggests that this measurement of self-mutilation in a chimpanzee can be used as an important auxiliary tool to evaluate de conditions of adaptation of an animal in captivity, functioning as a direct indicator of the presence of chronic stress.

Coupling of palmitate to ovalbumin inhibits the induction of oral tolerance

Oral tolerance is a phenomenon that may occur in animals exposed to protein antigens for the first time by the oral route. They become unable to produce immune responses at the levels normally observed when they are immunized parenterally with antigen in the presence of adjuvants. Lipids have been used as adjuvants for both parenteral and oral immunization. In the present study we coupled ovalbumin with palmitate residues by incubating the protein with the N-hydroxysuccinimide palmitate ester and tested the preparation for its ability to induce oral tolerance. This was performed by giving 20 mg of antigen to mice by the oral route 7 days prior to parenteral immunization in the presence of Al(OH)3. Mice were bled one week after receiving a booster that was given 2 weeks after primary immunization. Specific antibodies were detected by ELISA. Despite the fact that the conjugates are as immunogenic as the unmodified protein when parenterally injected in mice, they failed to induce oral tolerance. This discrepancy could be explained by differences in the intestinal absorption of the two forms of the antigen. In fact, when compared to the non-conjugated ovalbumin, a fast and high absorption of the lipid-conjugated form of ovalbumin was observed by "sandwich" ELISA.

Indirect effects of oral tolerance to ovalbumin interfere with the immune responses triggered by Schistosoma mansoni eggs

The objective of the present study was to investigate whether the injection of a tolerated protein (indirect effects) affects the formation of granulomas around Schistosoma mansoni eggs trapped in the lungs after intravenous (iv) injection into normal (noninfected) C57BL/6 mice (6 animals per group). To induce oral tolerance to chicken egg ovalbumin a 1/5 dilution of egg white in water was offered ad libitum in a drinking bottle for 3 days. Control mice received water. After 7 days, control and experimental animals were injected iv with 2,000 S. mansoni eggs through a tail vein. In some mice of both groups the iv injection of eggs was immediately followed by intraperitoneal (ip) immunization with 10 µg of dinitrophenylated conjugates of ovalbumin (DNP-Ova) emulsified in complete Freund's adjuvant (CFA) or only CFA; 18 days later, mice were bled and killed by ether inhalation. The lungs were fixed in formalin and embedded in paraffin. Serial sections of 5 µm were stained with Giemsa, Gomori's silver reticulin and Sirius red (pH 10.2). Granuloma diameters were measured in histological sections previously stained with Gomori's reticulin. Anti-DNP and anti-soluble egg antigen (SEA) antibodies were analyzed by ELISA. In mice orally tolerant to ovalbumin the concomitant ip injection of DNP-Ova resulted in significantly lower anti-SEA antibodies (ELISA*: 1395 ± 352 in non-tolerant and 462 ± 146 in tolerant mice) and affected granuloma formation around eggs, significantly decreasing granuloma size (area: 22,260 ± 2478 to 12,993 ± 3242 µm²). Active mechanisms triggered by injection of tolerated antigen (ovalbumin) reduce granuloma formation.

Induction of oral tolerance and the effect of interleukin-4 on murine skin allograft rejection

We studied the effect of oral and portal vein administration of alloantigens on mouse skin allograft survival. Graft receptor BALB/c mice received spleen cells (30, 90, 150 or 375 x 10(6)) from donor C57BL/6 mice intragastrically on three successive days, starting seven days before the skin graft. Allograft survival was significantly increased with the feeding of 150 x 10(6) allogeneic spleen cells by one gavage (median survival of 12 vs 14 days, P <= 0.005) or when 300 x 10(6) cells were given in six gavage (12 vs 14 days, P < 0.04). A similar effect was observed when 150 x 10(6) spleen cells were injected into the portal vein (12 vs 14 days, P <= 0.03). Furthermore, prolonged allograft survival was observed with subcutaneous (12 vs 16 days, P <= 0.002) or systemic (12 vs 15 days, P <= 0.016) application of murine interleukin-4 (IL-4), alone or in combination with spleen cell injection into the portal vein (12 vs 18 days, P <= 0.0018). Taken together, these results showed that tolerance induction with spleen cells expressing fully incompatible antigens by oral administration or intraportal injection partially down-modulates skin allograft rejection. Furthermore, these findings demonstrated for the first time the effect of subcutaneous or systemic IL-4 application on allograft skin survival suggesting its use as a beneficial support therapy in combination with a tolerance induction protocol.

The time course of ethanol tolerance: associative learning

The effect of different contextual stimuli on different ethanol-induced internal states was investigated during the time course of both the hypothermic effect of the drug and of drug tolerance. Minimitters were surgically implanted in 16 Wistar rats to assess changes in their body temperature under the effect of ethanol. Rat groups were submitted to ethanol or saline trials every other day. The animals were divided into two groups, one receiving a constant dose (CD) of ethanol injected intraperitoneally, and the other receiving increasing doses (ID) during the 10 training sessions. During the ethanol training sessions, conditioned stimuli A (tone) and B (buzzer) were presented at "state +" (35 min after drug injection) and "state -" (170 min after drug injection), respectively. Conditioned stimuli C (bip) and D (white noise) were presented at moments equivalent to stimuli A and B, respectively, but during the saline training sessions. All stimuli lasted 15 min. The CD group, but not the ID group, developed tolerance to the hypothermic effect of ethanol. Stimulus A (associated with drug "state +") induced hyperthermia with saline injection in the ID group. Stimulus B (associated with drug "state -") reduced ethanol tolerance in the CD group and modulated the hypothermic effect of the drug in the ID group. These results indicate that contextual stimuli acquire modulatory conditioned properties that are associated with the time course of both the action of the drug and the development of drug tolerance.

Influence of the moisture at harvest and drying process of the grains on the level of carotenoids in maize (Zea mays)

AbstractMaize is considered a source of carotenoids; however, these compounds are highly unstable, degraded by high temperatures, exposure to light and presence of oxygen. The objective of this work was to evaluate the influence of the moisture and type of drying applied to grains on the level of carotenoids in yellow maize. The experiment was conducted in a completely randomized design (2 × 4 factorial), two levels of initial moisture at the harvest (22 and 19%) and three types of drying (in the sun; in the shade and in a dryer) and control (no drying). The samples of grains after drying with 12% of final moisture were analyzed by concentration of total carotenoids, carotenes (α-carotene + β-carotene), monohydroxilated carotenoids (β-cryptoxanthin), and xanthophylls (lutein + zeaxanthin). Initial moisture, type of drying and the interaction between moisture versus drying influence (p≤0.05) the levels of carotenoids in grains. This is the first report about the drying conditions and harvest’s initial moisture as influence on the profile and content of carotenoids in maize grains. Based on the results, this work suggested that the harvest be carried out preferably when the grains present 22% humidity, with drying in a dryer or in shade for further use or storage.

Poverty levels and children's health status: study of risk factors in an urban population of low socioeconomic level

To test the hypothesis that the low socioeconomic population living is shanty towns in Porto Alegre presents different levels of poverty which are reflected on its health status, a cross-sectional study was designed involving 477 families living in Vila Grande Cruzeiro, Porto Alegre, Brazil. The poverty level of the families was measured by using an instrument specifically designed for poor urban populations. Children from families living in extreme poverty (poorest quartile) were found to have higher infant mortality rate, lower birth weights, more hospitalizations, and higher malnutrition rates, in addition to belonging to more numerous families. Thus, the shanty town population of Porto Alegre is not homogeneous, and priority should be given to the more vulnerable subgroups.

The effect of Schistosoma mansoni infection on child morbidity in the State in Bahia, Brazil: II — Analysis at the individual level

The present investigation was carried out on a sample of 840 children (5 to 16 years old) from ten small towns of the State of Bahia in northeastern Brazil. The objetive was to study, by using a cross sectional methodology, the evolution of schistosomiasis morbidity (hepatic and splenic enlargement) in children, and the role of the intensity of S. mansoni infection in this process. The children were analised in three age groups (5 to 8, to 12 and 13 to 16 years old) and classified as uninfected, mildly infected, moderately infected and heavily infected according to the number of eggs in the stool. In children aged 5 to 8 years, increasing egg counts were not associated with increasing frequencies of hepatic or splenic enlargement. In the 9 to 12 years old group and association was observed with the prevalence of hepatic enlargement, but not with the prevalence of spleen enlargement. In the oldest group, 13 to 16 years old, an association was observed with the prevalence of enlargement of both organs. It was evident that in this population schistosomiasis morbidity develops in the early period of life as a gradual process starting with liver enlargement and followed by spleen enlargement some years later. It was found that the intensity of infection has a fundamental role in this process, although there is a latent period of some years before clinical splenomegaly appears in moderate-heavily infected children. The Authors suggest that the prevalence of splenomegaly in the 13 to 16 years old group is a good measure of the community level of schistosomiasis morbidity and could be used to measure the impact of control programs.

Reduced capacity of peritoneal exudate cells for oxidative killing of Schistosoma mansoni schistosomula

Peritoneal exudate cells from mice infected with Schistosoma mansoni (S-PEC) can kill schistosomula in vitro in the presence of immune serum. S-PEC produce a low level of respiratory burst, and schistosomula mortality in their presence is not reduced when exogenous antioxidants are added, suggesting that with S-PEC, oxidative killing is not important. Hydrogen peroxide (H2O2) and superoxide production by S-PEC, and cells from BCG and thioglycollate (THGL) injected non-infected mice, non-specifically stimulated with opsonized zymosan, were measured. Levels of H2O2 produced by S-PEC were significantly lower than BCG or THGL PEC, and were below the H2O2 threshold for schistosomula killing. This resulted in lower levels of cell-mediated killing of schistosomula in vitro by S-PEC than by BCG or THGL PEC. Superoxide levels, however, were similar between the three cell populations. The efficiency of PEC to kill schistosomules in vitro correlated with H2O2 rather than superoxide levels. The lower tolerance of schistosomula, compared to adult S. mansoni to GSH depleting agents increases their sensitivity to oxidative attack and resulted in higher levels of cell-mediated killing in vitro.