Influência de estratégia de educação em saúde mediada por álbum seriado sobre a autoeficácia materna para amamentar / Influence of health education strategy mediated by a self-efficacy breastfeeding serial album / Influencia de estrategia de educación en salud mediada por álbum ilustrado acerca de la autoeficacia materna para amamantar

Objetivou-se verificar a autoeficácia da puérpera em amamentar, antes e após a intervenção educativa. Optou-se por um estudo quantitativo, desenvolvido com 100 puérperas de uma maternidade pública de Fortaleza-CE. Antes e após aplicação do álbum seriado "Eu posso amamentar o meu filho", foram realizadas entrevistas utilizando dois formulários abordando dados de identificação da puérpera e a Breastfeeding Self-Efficacy Scale Short Form. Constatou-se um aumento dos escores da escala após a utilização da intervenção educativa, sobretudo em puérperas com características específicas, já que se observou associação estatisticamente significante entre as seguintes variáveis: idade entre 20-29 anos; estado civil casado/união consensual; número de cinco a sete moradores na casa; puérperas que exerciam atividades remuneradas fora do lar e rendas familiares de dois a oito salários mínimos. Conclui-se que a tecnologia educativa implementada às puérperas foi eficaz no aumento da autoeficácia materna em amamentar, podendo resultar, consequentemente, no alcance de boas taxas de aleitamento materno.

The objective was to verify the self-efficacy of new mothers before and after an educational intervention. We opted for a quantitative study involving 100 new mothers of a public maternity hospital in Fortaleza-CE, Brazil. Before and after the application of the serial album "I can breastfeed my child", interviews were performed using two forms: the first was used to collect identification data of the participants and the second was the Breastfeeding Self-Efficacy Scale Short Form. We found an increase in scores on the scale after the educational intervention, particularly in mothers with specific characteristics, with a statistically significant association between the following variables: age between 20 and 29 years; married/consensual union; from five to seven household members; engaged in paid work outside the home; and family income from two to eight minimum wages. We concluded that the educational intervention implemented with women who had just given birth was effective in increasing maternal breastfeeding self-efficacy, which may result in improving breastfeeding rates.

El objetivo fue verificar la autoeficacia de puérperas en amamantar, antes y después de la intervención educativa. Estudio cuantitativo, con 100 mujeres de maternidad pública de Fortaleza-CE, Brasil. Antes y después de la aplicación del álbum ilustrado "Yo puedo amamantar a mi hijo", fueron realizadas entrevistas con dos encuestas para abordar datos de identificación de la puérpera y Breastfeeding Self-Efficacy Scale Short Form. Hubo aumento en las puntuaciones de la escala después de la intervención educativa, especialmente en las madres con características específicas, con asociación estadísticamente significativa entre las variables: edades entre 20-29 años; estado civil casado/unión libre; número de cinco a siete miembros del hogar; madres que realizaban trabajo remunerado fuera del hogar; rentas familiares de dos a ocho sueldos mínimos. La tecnología educativa implementada a las puérperas fue eficaz cuanto al aumento de la autoeficacia materna en amamantar, resultando, por lo tanto, en el alcance de buenos índices de lactancia materna.

A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni

Schistosomiasis is a major neglected tropical disease caused by trematodes from the genus Schistosoma. Because schistosomes exhibit a complex life cycle and numerous mechanisms for regulating gene expression, it is believed that spliced leader (SL) trans-splicing could play an important role in the biology of these parasites. The purpose of this study was to investigate the function of trans-splicing in Schistosoma mansoni through analysis of genes that may be regulated by this mechanism and via silencing SL-containing transcripts through RNA interference. Here, we report our analysis of SL transcript-enriched cDNA libraries from different S. mansoni life stages. Our results show that the trans-splicing mechanism is apparently not associated with specific genes, subcellular localisations or life stages. In cross-species comparisons, even though the sets of genes that are subject to SL trans-splicing regulation appear to differ between organisms, several commonly shared orthologues were observed. Knockdown of trans-spliced transcripts in sporocysts resulted in a systemic reduction of the expression levels of all tested trans-spliced transcripts; however, the only phenotypic effect observed was diminished larval size. Further studies involving the findings from this work will provide new insights into the role of trans-splicing in the biology of S. mansoni and other organisms. All Expressed Sequence Tags generated in this study were submitted to dbEST as five different libraries. The accessions for each library and for the individual sequences are as follows: (i) adult worms of mixed sexes (LIBEST_027999: JZ139310 - JZ139779), (ii) female adult worms (LIBEST_028000: JZ139780 - JZ140379), (iii) male adult worms (LIBEST_028001: JZ140380 - JZ141002), (iv) eggs (LIBEST_028002: JZ141003 - JZ141497) and (v) schistosomula (LIBEST_028003: JZ141498 - JZ141974).

Genetic diversity of Leishmania infantum field populations from Brazil

Leishmania infantum (syn. Leishmania chagasi) is the etiological agent of visceral leishmaniasis (VL) in Brazil. The epidemiology of VL is poorly understood. Therefore, a more detailed molecular characterization at an intraspecific level is certainly needed. Herein, three independent molecular methods, multilocus microsatellite typing (MLMT), random amplification of polymorphic DNA (RAPD) and simple sequence repeats-polymerase chain reaction (SSR-PCR), were used to evaluate the genetic diversity of 53 L. infantum isolates from five different endemic areas in Brazil. Population structures were inferred by distance-based and Bayesian-based approaches. Eighteen very similar genotypes were detected by MLMT, most of them differed in only one locus and no correlation was found between MLMT profiles, geographical origin or the estimated population structure. However, complex profiles composed of 182 bands obtained by both RAPD and SSR-PCR assays gave different results. Unweighted pair group method with arithmetic mean trees built from these data revealed a high degree of homogeneity within isolates of L. infantum. Interestingly, despite this genetic homogeneity, most of the isolates clustered according to their geographical origin.

Biological, biochemical and molecular features of Trypanosoma cruzi strains isolated from patients infected through oral transmission during a 2005 outbreak in the state of Santa Catarina, Brazil: its correspondence with the new T. cruzi Taxonomy Consensus (2009)

We examined strains of Trypanosoma cruzi isolated from patients with acute Chagas disease that had been acquired by oral transmission in the state of Santa Catarina, Brazil (2005) and two isolates that had been obtained from a marsupial (Didelphis aurita) and a vector (Triatoma tibiamaculata). These strains were characterised through their biological behaviour and isoenzymic profiles and genotyped according to the new Taxonomy Consensus (2009) based on the discrete typing unities, that is, T. cruzi genotypes I-VI. All strains exhibited the biological behaviour of biodeme type II. In six isolates, late peaks of parasitaemia, beyond the 20th day, suggested a double infection with biodemes II + III. Isoenzymes revealed Z2 or mixed Z1 and Z2 profiles. Genotyping was performed using three polymorphic genes (cytochrome oxidase II, spliced leader intergenic region and 24Sα rRNA) and the restriction fragment length polymorphism of the kDNA minicircles. Based on these markers, all but four isolates were characterised as T. cruzi II genotypes. Four mixed populations were identified: SC90, SC93 and SC97 (T. cruzi I + T. cruzi II) and SC95 (T. cruzi I + T. cruzi VI). Comparison of the results obtained by different methods was essential for the correct identification of the mixed populations and major lineages involved indicating that characterisation by different methods can provide new insights into the relationship between phenotypic and genotypic aspects of parasite behaviour.

Trypanosoma cruzi benznidazole susceptibility in vitro does not predict the therapeutic outcome of human Chagas disease

Therapeutic failure of benznidazole (BZ) is widely documented in Chagas disease and has been primarily associated with variations in the drug susceptibility of Trypanosoma cruzi strains. In humans, therapeutic success has been assessed by the negativation of anti-T. cruzi antibodies, a process that may take up to 10 years. A protocol for early screening of the drug resistance of infective strains would be valuable for orienting physicians towards alternative therapies, with a combination of existing drugs or new anti-T. cruzi agents. We developed a procedure that couples the isolation of parasites by haemoculture with quantification of BZ susceptibility in the resultant epimastigote forms. BZ activity was standardized with reference strains, which showed IC50 to BZ between 7.6-32 µM. The assay was then applied to isolates from seven chronic patients prior to administration of BZ therapy. The IC50 of the strains varied from 15.6 ± 3-51.4 ± 1 µM. Comparison of BZ susceptibility of the pre-treatment isolates of patients considered cured by several criteria and of non-cured patients indicates that the assay does not predict therapeutic outcome. A two-fold increase in BZ resistance in the post-treatment isolates of two patients was verified. Based on the profile of nine microsatellite loci, sub-population selection in non-cured patients was ruled out.

Differential tissue tropism of Trypanosoma cruzi strains: an in vitro study

We have previously demonstrated selection favoring the JG strain of Trypanosoma cruziin hearts of BALB/c mice that were chronically infected with an equal mixture of the monoclonal JG strain and a clone of the Colombian strain, Col1.7G2. To evaluate whether cell invasion efficiency drives this selection, we infected primary cultures of BALB/c cardiomyocytes using these same T. cruzi populations. Contrary to expectation, Col1.7G2 parasites invaded heart cell cultures in higher numbers than JG parasites; however, intracellular multiplication of JG parasites was more efficient than that of Col1.7G2 parasites. This phenomenon was only observed for cardiomyocytes and not for cultured Vero cells. Double infections (Col1.7G2 + JG) showed similar results. Even though invasion might influence tissue selection, our data strongly suggest that intracellular development is important to determine parasite tissue tropism.

Trypanosoma cruzi: ancestral genomes and population structure

Although the genome of Trypanosoma cruzi has been completely sequenced, little is known about its population structure and evolution. Since 1999, two major evolutionary lineages presenting distinct epidemiological characteristics have been recognised: T. cruzi I and T. cruzi II. We describe new and important aspects of the population structure of the parasite, and unequivocally characterise a third ancestral lineage that we propose to name T. cruzi III. Through a careful analysis of haplotypes (blocks of genes that are stably transmitted from generation to generation of the parasite), we inferred at least two hybridisation events between the parental lineages T. cruzi II and T. cruzi III. The strain CL Brener, whose genome was sequenced, is one such hybrid. Based on these results, we propose a simple evolutionary model based on three ancestral genomes, T. cruzi I, T. cruzi II and T. cruzi III. At least two hybridisation events produced evolutionarily viable progeny, and T. cruzi III was the cytoplasmic donor for the resulting offspring (as identified by the mitochondrial clade of the hybrid strains) in both events. This model should be useful to inform evolutionary and pathogenetic hypotheses regarding T. cruzi.

Functional complementation of a yeast knockout strain by Schistosoma mansoni Rho1 GTPase in the presence of caffeine, an agent that affects mutants defective in the protein kinase C signal transduction pathway

In a previous study, the Schistosoma mansoni Rho1 protein was able to complement Rho1 null mutant Saccharomyces cerevisiae cells at restrictive temperatures and under osmotic stress (low calcium concentration) better than the human homologue (RhoA). It is known that under osmotic stress, the S. cerevisiae Rho1 triggers two distinct pathways: activation of the membrane 1,3-beta-glucan synthase enzymatic complex and activation of the protein kinase C1 signal transduction pathway, promoting the transcription of response genes. In the present work the SmRho1 protein and its mutants smrho1E97P, smrho1L101T, and smrho1E97P, L101T were used to try to clarify the basis for the differential complementation of Rho1 knockout yeast strain by the human and S. mansoni genes. Experiments of functional complementation in the presence of caffeine and in the presence of the osmotic regulator sorbitol were conducted. SmRho1 and its mutants showed a differential complementation of the yeast cells in the presence of caffeine, since smrho1E97P and smrho1E97P, L101T mutants showed a delay in the growth when compared to the yeast complemented with the wild type SmRho1. However, in the presence of sorbitol and caffeine the wild type SmRho1 and mutants showed a similar complementation phenotype, as they allowed yeast growth in all caffeine concentrations tested.

Developmental effects of additional ultraviolet a radiation, growth regulators and tyrosine in Alternanthera brasiliana (L) Kuntze cultured in vitro

Culturas de tecidos de Alternanthera brasiliana (L.) Kuntze foram tratadas com diferentes reguladores de crescimento (Cinetina e 2,4-D), tirosina e com ultravioleta longo (UV-A; 320 -400 nm) adicional com o intuito de observar seus efeitos no desenvolvimento e produção de pigmentos. Segmentos nodais de plantas crescidas a partir de sementes foram inoculados nos meios de cultura testados e mantidos sob os diferentes tipos de iluminação. Após 8 semanas este material foi utilizado para avaliação da produção de biomassa, clorofilas e betacianinas. O meio de Murashige and Skoog (MS) + cinetina proporcionou plântulas com até 4 brotos/explante. Este meio iluminado com luz branca (tipo luz do dia) foi a combinação mais adequada para micropropagação, pois apresenta maior porcentagem de enraizamento e maior produção de betacianinas. Plântulas crescidas sob iluminação com ultravioleta adicional tiveram diminuídas tanto a produção de biomassa quanto a relação Clor a/ Clor b. A adição de 2,4-D ao meio de cultura resultou na inibição da produção de pigmentos e no crescimento das plântulas.

Trypanosoma cruzi: genetic structure of populations and relevance of genetic variability to the pathogenesis of chagas disease

Chagas disease, caused by the protozoan Trypanosoma cruzi, has a variable clinical course, ranging from symptomless infection to severe chronic disease with cardiovascular or gastrointestinal involvement or, occasionally, overwhelming acute episodes. The factors influencing this clinical variability have not been elucidated, but it is likely that the genetic variability of both the host and the parasite are of importance. In this work we review the the genetic structure of T. cruzi populations and analyze the importance of genetic variation of the parasite in the pathogenesis of the disease under the light of the histotropic-clonal model.

Usefulness of microsatellite typing in population genetic studies of Trypanosoma cruzi

Through microsatellite analysis of 53 monoclonal populations of Trypanosoma cruzi, we found a remarkable degree of genetic polymorphism with no single multilocus genotype being observed more than once. The microsatellite profile proved to be stable during 70 generations of the CL Brener clone in culture. The microsatellite profiling presented also high diagnostic sensitivity since DNA amplifications could be achieved with less than 100 fg DNA, corresponding to half parasite total DNA content. Based on these technical attributes the microsatellite assay turns out to be an important tool for direct typing T. cruzi in biological samples. By using this approach we were able to type T. cruzi in feces of artificially infected bugs and in single cells sorted by FACS. The microsatellites have shown to be excellent markers for T. cruzi phylogenetic reconstruction. We used maximum parsimony based on the minimum number of mutational steps to build an unrooted Wagner network, which confirms previous conclusions based on the analysis of the D7 domain of the LSU rDNA gene that T. cruzi is composed by two major groups. We also obtained evidence that strains belonging to rRNA group 2 are subdivided into two genetically distant clusters, and that one of these clusters is more related to rRNA group 1/2. These results suggest different origins for these strains

The population structure of Trypanosoma cruzi: expanded analysis of 54 strains using eight polymorphic CA-repeat microsatellites

Recently we cloned and sequenced the first eight Trypanosoma cruzi polymorphic microsatellite loci and studied 31 clones and strains to obtain valuable information about the population structure of the parasite. We have now studied 23 further strains, increasing from 11 to 31 the number of strains obtained from patients with chronic Chagas disease. This expanded set of 54 strains and clones analyzed with the eight microsatellites markers confirmed the previously observed diploidy, clonal population organization and very high polymorphism of T. cruzi. Moreover, this new study disclosed two new features of the population genetic structure of T. cruzi. The first was the discovery that, similarly to what we had previously shown for strains isolated from insect vectors, mammals and humans with acute disease, isolates from patients in the chronic phase of Chagas disease could also be multiclonal, albeit at a reduced proportion. Second, when we used parsimony to display the genetic relationship among the clonal lineages in an unrooted Wagner network we observed, like before, a good correlation of the tree topography with the classification in three clusters on the basis of single locus analysis of the ribosomal RNA genes. However, a significant new finding was that now the strains belonging to cluster 2 split in two distant sub-clusters. This observation suggests that the evolutionary history of T. cruzi may be more complex than we previously thought.

Effectes of high temperature on survival, symbiotic performance and genomic modifications of bean nodulating Rhizobium strains

High temperatures can affect the survival, establishment and symbiotic properties of "Rhizobium" strains. Bean nodulating "Rhizobium" strains are considered particularly sensitive because on this strains genetic recombinations and/or deletions occur frequently, thus compromising the use of these bacteria as inoculants. In this study "R. tropici" and "R. leguminosarum" bv. "phaseoli" strains isolated from Cerrado soils were exposed to thermal stress and the strains' growth, survival and symbiotic relationships as well as alterations in their genotypic and phenotypic were analysed. After successive thermal shocks at 45ºC for four hours, survival capacity appeared to be strain-specifc, independent of thermo-tolerance and was more apparent in "R. tropici" strains (with the exception of FJ2.21) were more stable than "R. leguminosarum" bv. "phaseoli" strains because no significant phenotypic alterations were observed following thermal treatments and they maintained their original genotypic pattern after innoculation in plants.

Biological parameters and molecular markers of clone CL brener - the reference organism of the Trypanosoma cruzi genome project

Clone CL Brener is the reference organism used in the Trypanosma cruzi Genome Project. Some biological paramenters of CL Brener were determined: (a) the doubling time of epimastigote forms cultured in liver infusion-tryptose (LIT) medium at 28ºC is 58ñ13 hr; (b) differentiation of epimastigotes to metacyclic trypomastigotes is obtained by incubation in LIT-20 per cent Grace's medium; (c) trypomastigotes infect mammalian cultured cells and perform the complete intracellular cycle at 33 and 37ºC; (c) blood forms are highly infective to mice; (e) blood forms are susceptible to nifurtimox and benznidazole. The molecular typing of CL Brener has been determined: (a) isoenzymatic profiles are characteristic of zymodeme ZB; (b) PCR amplification of a 24 alpha ribosomal RNA sequence indicates it belongs to T. cruzi lineage 1; (c) schizodeme, randomly amplified polymorphic DNA (RAPD) and DNA fingerprinting analyses were performed.

Paternity testing in Brazil by DNA fingerprinting with the multilocus probes F10

Empirical analysis of 200 paternity cases with the F10 multilocus DNA fingerprinting probe showed that it was capable of distinguishing fathers from nonfathers in every case. The average exclusion probability was0.99998. A very effective discrimination parameter was the proportion of nonmaternal (test) bands which cannot be detected in the alleged father (unassignable bands) among all test bands. Values below 0.2 were seen in true fathers while in all cases of nonfathers the values were above 0.35. Minisatellite mutations occurred at a rate of 0.004 per band per child. The distribution of band-sharing among first degree relatives and unrelated individuals showed only a small overlap. Thus, band-sharing of the F10 fingerprints should provide a useful statistics for testing genetic relationships in deficiency cases.