Antibody isotype responses in Balb/c mice immunized with the cytoplasmic repetitive antigen and flagellar repetitive antigen of Trypanosoma cruzi

In the present report we analyzed the levels of IgG1, IgG2a, IgG2b and IgG3 isotypes from Balb/c mice immunized with cytoplasmic repetitive antigen (CRA), and flagelar repetitive antigen (FRA) of Trypanosoma cruzi. The immunization was done by subcutaneous route three times (20 days apart) and the analysis was performed 14 days after each treatment. CRA-immunized mice produced high levels of all IgG isotypes, mainly IgG3 and IgG1. FRA-immunization elicited only high levels of IgG1.

Mycobacterium avium restriction fragment lenght polymorphism-IS IS1245 and the simple double repetitive element polymerase chain reaction typing method to screen genetic diversity in Brazilian strains

Simple double repetitive element polymerase chain reaction (MaDRE-PCR) and Pvu II-IS1245 restriction fragment length polymorphism (RFLP) typing methods were used to type 41 Mycobacterium avium isolates obtained from 14 Aids inpatients and 10 environment and animals specimens identified among 53 mycobacteria isolated from 237 food, chicken, and pig. All environmental and animals strains showed orphan patterns by both methods. By MaDRE-PCR four patients, with multiple isolates, showed different patterns, suggesting polyclonal infection that was confirmed by RFLP in two of them. This first evaluation of MaDRE-PCR on Brazilian M. avium strains demonstrated that the method seems to be useful as simple and less expensive typing method for screening genetic diversity in M. avium strains on selected epidemiological studies, although with limitation on analysis identical patterns except for one band.

CA88, a nuclear repetitive DNA sequence identified in Schistosoma mansoni, aids in the genotyping of nine Schistosoma species of medical and veterinary importance

CA88 is the first long nuclear repetitive DNA sequence identified in the blood fluke, Schistosoma mansoni. The assembled S. mansoni sequence, which contains the CA88 repeat, has 8,887 nucleotides and at least three repeat units of approximately 360 bp. In addition, CA88 also possesses an internal CA microsatellite, identified as SmBr18. Both PCR and BLAST analysis have been used to analyse and confirm the CA88 sequence in other S. mansoni sequences in the public database. PCR-acquired nuclear repetitive DNA sequence profiles from nine Schistosoma species were used to classify this organism into four genotypes. Included among the nine species analysed were five sequences of both African and Asian lineages that are known to infect humans. Within these genotypes, three of them refer to recognised species groups. A panel of four microsatellite loci, including SmBr18 and three previously published loci, has been used to characterise the nine Schistosoma species. Each species has been identified and classified based on its CA88 DNA fingerprint profile. Furthermore, microsatellite sequences and intra-specific variation have also been observed within the nine Schistosoma species sequences. Taken together, these results support the use of these markers in studying the population dynamics of Schistosoma isolates from endemic areas and also provide new methods for investigating the relationships between different populations of parasites. In addition, these data also indicate that Schistosoma magrebowiei is not a sister taxon to Schistosoma mattheei, prompting a new designation to a basal clade.

Comparison of spoligotyping, mycobacterial interspersed repetitive units typing and IS6110-RFLP in a study of genotypic diversity of Mycobacterium tuberculosis in Delhi, North India

The aim of the present study was to compare polymerase chain reaction (PCR)-based methods - spoligotyping and mycobacterial interspersed repetitive units (MIRU) typing - with the gold-standard IS6110 restriction fragment length polymorphism (RFLP) analysis in 101 isolates of Mycobacterium tuberculosis to determine the genetic diversity of M. tuberculosis clinical isolates from Delhi, North India. Spoligotyping resulted in 49 patterns (14 clusters); the largest cluster was composed of Spoligotype International Types (SITs)26 [Central-Asian (CAS)1-Delhi lineage], followed by SIT11 [East-African-Indian (EAI) 3-Indian lineage]. A large number of isolates (75%) belonged to genotypic lineages, such as CAS, EAI and Manu, with a high specificity for the Indian subcontinent, emphasising the complex diversity of the phylogenetically coherent M. tuberculosis in North India. MIRU typing, using 11 discriminatory loci, was able to distinguish between all but two strains based on individual patterns. IS6110-RFLP analysis (n = 80 strains) resulted in 67 unique isolates and four clusters containing 13 strains. MIRUs discriminated all 13 strains, whereas spoligotyping discriminated 11 strains. Our results validate the use of PCR-based molecular typing of M. tuberculosis using repetitive elements in Indian isolates and demonstrate the usefulness of MIRUs for discriminating low-IS6110-copy isolates, which accounted for more than one-fifth of the strains in the present study.

Coxiella burnetii, the agent of Q fever in Brazil: its hidden role in seronegative arthritis and the importance of molecular diagnosis based on the repetitive element IS1111 associated with the transposase gene

Coxiella burnetii is the agent of Q fever , an emergent worldwide zoonosis of wide clinical spectrum. Although C. burnetii infection is typically associated with acute infection, atypical pneumonia and flu-like symptoms, endocarditis, osteoarticular manifestations and severe disease are possible, especially when the patient has a suppressed immune system; however, these severe complications are typically neglected. This study reports the sequencing of the repetitive element IS1111 of the transposase gene of C. burnetii from blood and bronchoalveolar lavage (BAL) samples from a patient with severe pneumonia following methotrexate therapy, resulting in the molecular diagnosis of Q fever in a patient who had been diagnosed with active seronegative polyarthritis two years earlier. To the best of our knowledge, this represents the first documented case of the isolation of C. burnetii DNA from a BAL sample.

The remarkable journey of adaptation of the Plasmodium falciparum malaria parasite to New World anopheline mosquitoes

Plasmodium falciparum originated in Africa, dispersed around the world as a result of human migration and had to adapt to several different indigenous anopheline mosquitoes. Anophelines from the New World are evolutionary distant form African ones and this probably resulted in a more stringent selection of Plasmodium as it adapted to these vectors. It is thought that Plasmodium has been genetically selected by some anopheline species through unknown mechanisms. The mosquito immune system can greatly limit infection and P. falciparum evolved a strategy to evade these responses, at least in part mediated by Pfs47, a highly polymorphic gene. We propose that adaptation of P. falciparum to new vectors may require evasion of their immune system. Parasites with a Pfs47 haplotype compatible with the indigenous mosquito vector would be able to survive and be transmitted. The mosquito antiplasmodial response could be an important determinant of P. falciparum population structure and could affect malaria transmission in the Americas.

Translation and cultural adaptation of the Perceived Stigmatization Questionnaire for burn victims in Brazil

This methodological study aimed to describe the process of translation and cultural adaptation of the Perceived stigmatization Questionnaire (PSQ) and analyze the internal consistency of the items in the step of pre-testing. The PSQ was developed to evaluate the perception of stigmatizing behaviors of burn victims. The adaptation process was carried out from August 2012 to February 2013, comprising the steps outlined in the literature. As part of this process, the pre-test with 30 adult burn victims was held. All participants at this step reported to understand the instrument items and the scale of responses. There were no suggestions or changes in the tested version. The value of Cronbach’s alpha at pre-test was 0.87. The contribution of this study is to describe the operation of each of the steps of this methodological process and show the internal consistency of the items in the pre-test.

Cultural adaptation of the scale Pain Assessment in Advanced Dementia – PAINAD to Brazil

Objective: To translate and culturally adapt to Brazil the scale Pain Assessment in Advanced Dementia(PAINAD).Method: The cultural adaptation process followed the methodology of a theorical reference, in five steps: translation to Brazilian Portuguese, consensual version of translations, back-translation to the original language, revision by a committee of specialists in the field and a equivalency pre-test. The instrument was assessed and applied by 27 health professionals in the last step. Results: The Escala de Avaliação de Dor em Demência Avançada was culturally adapted to Brazil and presented semantic equivalency to the original, besides clarity, applicability and easy comprehension of the instrument items. Conclusion: This process secured the psychometric properties as the reliability and content validity of the referred scale.


Study of adaptation and validation of the Practice environment scale of the nursing work index for the portuguese reality

Objective: Testing the psychometric properties of the Portuguese version of the Practice Environment Scale of the Nursing Work Index. Method: A descriptive, analytical and cross-sectional study, for the cross-cultural adaptation and validation of the psychometric properties of the scale. The study participants were 236 nurses from two hospitals in the regions of Lisbon and Vale do Tejo. Results: The 0.92 Cronbach’s alpha was obtained for overall reliability and support of a five-dimension structure. Conclusion: The excellent quality of adjustment of analysis confirms the validity of the adapted version to hospital care settings, although there was no total coincidence of items in the five dimensions


Evaluation of professional knowledge and attitudes on dementia patient care: a trans-cultural adaptation of an evaluation instrument

Objective To describe the trans-cultural adaptation of the evaluation instrument entitled Atenció Sanitària de Les Demències: la visió de L' Atenció Primarià from Catalan into versions in Portuguese for doctors and nurses. This study evaluates the knowledge and perspectives of these professionals in their treatment of patients diagnosed with dementia in cases of primary care. Method The adaptation followed internationally accepted rules, which include the following steps: translation, synthesis, back-translation, revision by a committee of specialists, and a test run with 35 practicing doctors and 35 practicing nurses in Brazil's Family Health Strategy (Estratégia Saúde da Família, or ESF in Portuguese). Results The translation, synthesis, and back-translation steps were performed satisfactorily; only small adjustments were required. The committee of specialists verified the face validity in the version translated into Portuguese, and all of the items that received an agreement score lower than 80% during the initial evaluation were revised. In the test run, the difficulties presented by the health care professionals did not reach 15% of the sample, and therefore, no changes were made. Conclusion The Portuguese translation of the instrument can be considered semantically, idiomatically, culturally, and conceptually equivalent to the original Catalan version and is, therefore, appropriate for use in Brazil.

Cross-cultural adaptation of the Quality of Life Index Spinal Cord Injury - Version III

OBJECTIVE To translate and culturally adapt to Portuguese the Ferrans and Powers Quality of Life Index Spinal Cord Injury - Version III and characterize the sample in relation to sociodemographic and clinical aspects. METHOD A methodological study with view to cross-cultural adaptation, following the particular steps of this method: initial translation, translation synthesis, back-translation (translation back to the original language), review by a committee of judges and pretest of the final version. The pretest was carried out with 30 patients with spinal cord injury. RESULTS An index of 74 items divided into two parts (satisfaction/importance) was obtained. The criteria of semantic equivalence were evaluated as very adequate translation, higher than 87%, and vocabulary and were grammar higher than 86%. Idiomatic equivalence was higher than 74%, experimental greater than 78% and conceptual was greater than 70%. CONCLUSION After cross-cultural adaptation, the instrument proved semantic, idiomatic, experimental and conceptual adequacy, in addition to helping the evaluation of the quality of life of people with spinal cord injury.

Cross-cultural adaptation of the Caregiver Reaction Assessment for use in Brazil with informal caregivers of the elderly

This study aimed to carry out the cross-cultural adaptation of the Caregiver Reaction Assessment CRA for use in Brazil with informal caregivers of dependent elderly METHOD A methodological study, of five steps: initial translation, synthesis of translations, retro-translation, evaluation by a judge committee and a pre-test, with 30 informal caregivers of older persons in Fortaleza, Brazil. Content validity was assessed by five experts in gerontology and geriatrics. The cross-cultural adaptation was rigorously conducted, allowing for inferring credibility. RESULTS The Brazilian version of the CRA had a simple and fast application (ten minutes), easily understood by the target audience. It is semantically, idiomatically, experimentally and conceptually equivalent to the original version, with valid content to assess the burden of informal caregivers for the elderly (Content Validity Index = 0.883). CONCLUSION It is necessary that other psychometric properties of validity and reliability are tested before using in care practice and research.

Leaf area of common bean genotypes during early pod filling as related to plant adaptation to limited phosphorus supply

Low phosphorus supply markedly limits leaf growth and genotypes able to maintain adequate leaf area at low P could adapt better to limited-P conditions. This work aimed to investigate the relationship between leaf area production of common bean (Phaseolus vulgaris) genotypes during early pod filling and plant adaptation to limited P supply. Twenty-four genotypes, comprised of the four growth habits in the species and two weedy accessions, were grown at two P level applied to the soil (20 and 80 mg kg-1) in 4 kg pots and harvested at two growth stages (pod setting and early pod filling). High P level markedly increased the leaf number and leaf size (leaf area per leaf), slightly increased specific leaf area but did not affect the net assimilation rate. At low P level most genotypic variation for plant dry mass was associated with leaf size, whereas at high P level this variation was associated primarily with the number of leaves and secondarily with leaf size, specific leaf area playing a minor role at both P level. Determinate bush genotypes presented a smaller leaf area, fewer but larger leaves with higher specific leaf area and lower net assimilation rate. Climbing genotypes showed numerous leaves, smaller and thicker leaves with a higher net assimilation rate. Indeterminate bush and indeterminate prostrate genotypes presented the highest leaf area, achieved through intermediate leaf number, leaf size and specific leaf area. The latter groups were better adapted to limited P. It is concluded that improved growth at low P during early pod filling was associated with common bean genotypes able to maintain leaf expansion through leaves with greater individual leaf area.

Performance of food-type soybean genotypes and their possibility for adaptation to Brazilian latitudes

This work was conducted at the Universidade Estadual de Londrina (UEL), in Londrina, State of Paraná, Brazil, with the goal to study food-type soybean (Glycine max (L.) Merrill) genotypes performance for use in cultivation or crosses. A total of 104 genotypes were analyzed: 88 were food-type with large seeds, eight were food-type with small seeds, and eight-grain types adapted cultivars. The experimental plan was in randomized complete block design with four replications, and 12 traits of agronomic importance were considered. Genetic diversity was observed in the food-type germplasm. There were some genotypes with high yield adapted to a normal period of sowing. Soybean genetic improvement programs for direct human consumption in Brazil, either by means of Asiatic pure lines or by means of the incorporation of genes for late flowering in short-day conditions in this lines is highly viable.