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.

Excesso de peso e hipertensão arterial em trabalhadores de empresas beneficiadas pelo Programa de Alimentação do Trabalhador (PAT)

OBJETIVO: Avaliar as prevalências de excesso de peso (EP), hipertensão arterial (HA) e fatores associados em trabalhadores de empresas beneficiadas pelo Programa de Alimentação do Trabalhador (PAT) da cidade de São Paulo. MÉTODOS: Estudo transversal com 1.339 trabalhadores de 30 empresas. A coleta de dados envolveu a aplicação de um questionário com dados de caracterização dos trabalhadores e peso e altura auto-referidos. Foi realizada a aferição da pressão arterial e o estado nutricional foi classificado segundo o Índice de Massa Corporal (IMC). Odds ratios foram estimadas na avaliação dos fatores de risco associados a HA e EP. RESULTADOS: Os trabalhadores apresentaram, em média, 36,4 anos (dp = 10,3) e 9,9 anos de estudo (dp = 2,3), sendo 60% da amostra pertencente ao sexo masculino. Na comparação com homens, mulheres apresentaram valores significativamente menores de idade, pressão arterial sistólica (PAS) e diastólica (PAD) e IMC e maior escolaridade. As prevalências em homens de EP (25 kg/m2) (56%) e de HA (PAS > 140 mmHg e/ou PAD > 90 mmHg ou uso de medicações anti-hipertensivas) (38%), foram aproximadamente o dobro da registrada em mulheres (30% e 19%), respectivamente. Idade foi fator de risco para a ocorrência de EP e HA em ambos os sexos, enquanto que a escolaridade foi fator de proteção para EP e HA em mulheres e fator de risco para o desenvolvimento de EP em homens. CONCLUSÃO: Os trabalhadores do sexo masculino constituíram uma população de maior risco para ocorrência de HA e EP e devem ser priorizados nos programas que visam a prevenção dessas doenças. Neste sentido, o PAT pode representar um lugar de destaque nas ações de saúde e nutrição no ambiente de trabalho.

Tuberculose bovina no Estado da Paraíba: estudo retrospectivo

O objetivo do presente trabalho foi determinar a frequência de propriedades positivas (focos) e de animais positivos para a tuberculose bovina no Estado da Paraíba. Foram utilizados dados da Agência de Defesa Agropecuária do Estado, coletados de suas 23 microrregiões, durante o período de janeiro de 2008 a julho de 2009. Durante esse período, foram examinadas 10.963 propriedades e 54.472 bovinos foram submetidos ao teste de tuberculinização. Para o diagnóstico foi utilizada, como prova de triagem, a tuberculinização cervical simples para gado de leite e a tuberculinização na prega caudal para gado de corte; como prova confirmatória foi utilizada a tuberculinização cervical comparativa. Uma propriedade foi considerada foco quando apresentou pelo menos um animal soropositivo. Das propriedades investigadas, 62 (0,57%) apresentaram pelo menos um animal positivo e dos animais analisados, 136 (0,25%) foram positivos. Houve diferença significativa (p<0,001) na proporção de fêmeas (0,32%) e machos (0,04%) positivos. A despeito da baixa freqüência de focos de brucelose e de animais soropositivos, é necessária a condução de medidas que incluem a conscientização dos produtores, fiscalização nas barreiras sanitárias e levantamentos periódicos da situação epidemiológica desta doença, principalmente nas microrregiões com maior frequência da infecção, com o objetivo de evitar, ou pelo menos minimizar, a disseminação do agente.

Serosurvey of antibodies against spotted fever group Rickettsia spp. in horse farms in Northern Paraná, Brazil

Brazilian spotted fever (BSF) is an emerging disease most likely caused by Rickettsia rickettsii. The objective of the present study was to estimate the seroprevalence of BSF rickettsia infections in equines from six horse farms located in Londrina County, Paraná, Southern Brazil. Six owners of horse farms situated in Cambé, Santa Fé, Guaraci and Londrina municipalities participated in the study. All farms were located in areas where BSF has not been reported. A total of 273 horses were sampled and their sera were tested by indirect Immunofluorescence assay (IFA) using R. rickettsii and R. parkeri antigens. Titers equal to and greater than 64 were considered positive. Of 273 sera tested, 15 (5.5%) reacted to R. rickettsii and 5 (1.8%) to R. parkeri. Five out of the six farms studied revealed seropositive animals and seropositivity rate ranged from 0 to 13%. The titers ranged from 64 to 512, and four samples had a titer of 512. Nine animals reacted to R. rickettsii with titers four-fold higher than those for R. parkeri. These results suggest that horses in Northern Paraná may have been exposed to rickettsiae identical or closely related to R. rickettsii.

Rapid detection of bovine coronavirus by a semi-nested RT-PCR

Bovine coronavirus (BCoV) is a member of the group 2 of the Coronavirus (Nidovirales: Coronaviridae) and the causative agent of enteritis in both calves and adult bovine, as well as respiratory disease in calves. The present study aimed to develop a semi-nested RT-PCR for the detection of BCoV based on representative up-to-date sequences of the nucleocapsid gene, a conserved region of coronavirus genome. Three primers were designed, the first round with a 463bp and the second (semi-nested) with a 306bp predicted fragment. The analytical sensitivity was determined by 10-fold serial dilutions of the BCoV Kakegawa strain (HA titre: 256) in DEPC treated ultra-pure water, in fetal bovine serum (FBS) and in a BCoV-free fecal suspension, when positive results were found up to the 10-2, 10-3 and 10-7 dilutions, respectively, which suggests that the total amount of RNA in the sample influence the precipitation of pellets by the method of extraction used. When fecal samples was used, a large quantity of total RNA serves as carrier of BCoV RNA, demonstrating a high analytical sensitivity and lack of possible substances inhibiting the PCR. The final semi-nested RT-PCR protocol was applied to 25 fecal samples from adult cows, previously tested by a nested RT-PCR RdRp used as a reference test, resulting in 20 and 17 positives for the first and second tests, respectively, and a substantial agreement was found by kappa statistics (0.694). The high sensitivity and specificity of the new proposed method and the fact that primers were designed based on current BCoV sequences give basis to a more accurate diagnosis of BCoV-caused diseases, as well as to further insights on protocols for the detection of other Coronavirus representatives of both Animal and Public Health importance.

Associação da medroxiprogesterona ao protocolo Ovsynch para inseminação artificial em tempo fixo de búfalas cíclicas (Bubalus bubalis) criadas na Amazônia Oriental

O presente estudo visou avaliar os efeitos da associação da medroxiprogesterona (análogo sintético da progesterona) ao protocolo Ovsynch sobre o crescimento folicular, a ovulação e a taxa de concepção de búfalas criadas na Amazônia Oriental (Tracuateua-PA). Vinte e sete fêmeas adultas (G1 n=14 e G2 n=13), cíclicas, sem bezerro ao pé e com ECC 3,5 foram submetidas a Ovsynch. Os animais do G2 receberam 60 mg de medroxiprogesterona entre D0 e D7 (D0=início do tratamento). A ultra-sonografia ovariana foi realizada nos D 0, 7, 9 e 10. O contingente de folículos pequenos diferiu no D7 (G1: 4,57±0,60 versus G2: 6,54±0,67; P=0,05). Tempo e tratamento influenciaram o diâmetro folicular no D7. O crescimento do folículo dominante entre D7 e D9 foi maior nos animais tratados (G1: 2,05±0,49 mm/dia versus 3,48±0,41 mm/dia; P<0,05). Mais animais do G1 ovularam precocemente (35,71% versus 30,77%), porém isso não afetou as taxas de concepção (G1: 50,00% e G2: 30,77%; P>0,05). Os achados sugerem que a medroxiprogesterona (1) aumenta recrutamento folicular e retarda o crescimento dos folículos com diâmetro maior que 5,0 mm entre D0 e D7; (2) sua retirada incrementa em 1,7 vezes o crescimento folicular do D7 ao D9; (3) pode contribuir para a ovulação de folículos maiores e, em tese, para maior formação de tecido luteínico; (4) não promove ovulação precoce após o Ovsynch; (5) não eleva as taxas de concepção após sincronização de fêmeas cíclicas e com bom escore corporal, devendo ser avaliada para uso em fêmeas acíclicas ou com ECC mais baixo.

Violência física contra menores de 15 anos: estudo epidemiológico em cidade do sul do Brasil

OBJETIVO: apresentar o perfil de casos notificados de violência física contra menores de 15 anos em Londrina, Paraná, no ano de 2006. MÉTODO: Estudo transversal, com coleta retrospectiva nos prontuários dos Conselhos Tutelares e serviços de atendimento do município. Os dados foram processados e tabulados pelo programa Epi Info. RESULTADOS: Foram estudados 479 casos de violência por força corporal e 9 casos de violência por outros meios (7 por instrumentos, 1 por objeto cortante e 1 por substância corrosiva). Na violência por força corporal, predominaram vítimas do sexo feminino (53,4 por cento ) e maior risco na idade de seis anos (12,2 por 1.000). O pai foi o agressor mais frequente (48,8 por cento ) e o alcoolismo esteve presente em 64,0 por cento dos casos. A violência por instrumentos foi praticada através de cinta (42,9 por cento ), fio (28,6 por cento ), ferro (14,3 por cento ) e instrumento de cozinha (14,3 por cento ), com vítimas do sexo feminino (85,7 por cento ), na faixa etária de doze anos (33,3 por cento ), sendo o pai (71,4 por cento ) e a mãe (28,6 por cento ) os únicos agressores, com o alcoolismo presente em 57,1 por cento destas situações. A vítima de violência por objeto cortante era do sexo masculino, 13 anos e o agressor, desconhecido, tinha de 15 a 19 anos. A violência por substância corrosiva teve como vítima um adolescente de 13 anos, do sexo masculino, cujo agressor foi o pai, sendo o alcoolismo a situação presente. CONCLUSÕES: Os resultados apontam para a importância epidemiológica do abuso físico contra crianças e adolescentes e podem contribuir para a elaboração de estratégias de prevenção e acompanhamento das vítimas

Physical analysis of compost from composting plants in Brazil

Nowadays the composting process has shown itself to be an alternative in the treatment of municipal solid wastes by composting plants. However, although more than 50% of the waste generated by the Brazilian population is composed of matter susceptible to organic composting, this process is, still today, insufficiently developed in Brazil, due to low compost quality and lack of investments in the sector. The objective of this work was to use physical analyses to evaluate the quality of the compost produced at 14 operative composting plants in the Sao Paulo State in Brazil. For this purpose, size distribution and total inert content tests were done. The results were analyzed by grouping the plants according to their productive processes: plants with a rotating drum, plants with shredders or mills, and plants without treatment after the sorting conveyor belt. Compost quality was analyzed considering the limits imposed by the Brazilian Legislation and the European standards for inert contents. The size distribution tests showed the influence of the machinery after the sorting conveyer on the granule sizes as well as the inert content, which contributes to the presence of materials that reduce the quality of the final product

Association between neuromuscular tests and kumite performance on the Brazilian Karate National Team

The aim of this study was to verify the relationship of strength and power with performance on an international level karate team during official kumite simulations. Fourteen male black belt karate athletes were submitted to anthropometric data collection and then performed the following tests on two different days: vertical jump test, bench press and squat maximum dynamic strength (1RM) tests. We also tested power production for both exercises at 30 and 60% 1RM and performed a kumite match simulation. Blood samples were obtained at rest and immediately after the kumite matches to measure blood lactate concentration. Karate players were separated by performance (winners vs. defeated) on the kumite matches. We found no significant differences between winners and defeated for strength, vertical jump height, anthropometric data and blood lactate concentration. Interestingly, winners were more powerful in the bench press and squat exercises at 30% 1RM. Maximum strength was correlated with absolute (30% 1RM r = 0.92; 60% 1RM r = 0.63) and relative power (30% 1RM r = 0.74; 60% 1RM r = 0.11, p > 0.05) for the bench press exercise. We concluded that international level karate players' kumite match performance are influenced by higher levels of upper and lower limbs power production.

Systematic review of genus Cerradomys Weksler, percequillo and Voss, 2006 (Rodentia : Cricetidae : Sigmodontinae : Oryzomyini), with description of two new species from eastern Brazil

Cerradomy's is a monophyletic genus that includes four known species, Cerradomys subflavus, C maracajuensis, C. marinhus, and C. scotti, distributed throughout the open vegetation belt across South America, from northeastern Brazil to southeastern Bolivia, and from eastern to northwestern Paraguay. We revised the status of the species currently assigned to this genus by analyzing skins, skulls, karyotypes, and cytochrome b DNA sequences. We also described two novel species, one distributed in the Brazilian states of Minas Gerais, Bahia, and Sergipe, and the other in the states of Paraiba, Pernambuco, Piaui, Ceara, and Maranhao. Molecular analysis suggested the following phylogenetics arrangement: (((C. subflavus-C. sp.n.2) C. sp.n.1) C scotti)(C. marinhus-C. maracajuensis)). Apparently, both novel species inhabit the Caatinga domain and penetrated the coastal Atlantic rainforest, differing from the remaining congeneric species that are typical open-area inhabitants.

Evaluation of the spouted bed dried leaf extract of Bauhinia forficata for the treatment of experimental diabetes in rats

Previously, we have demonstrated that treatment of experimental diabetes with a decoction of Bauhinia forficata leaves is beneficial. In this study, we prepared a two-fold concentrate of this extract and tested its effects on physiological, biochemical and toxicity markers in streptozotocin-diabetic rats. Dried and ground leaves were extracted with warm 70% hydroethanol and the filtrate concentrated by evaporation at 50 degrees C. This solution was mixed with colloidal silicon dioxide (Tixosil-333 (R)) and dried in a spouted bed (BfT). Rats were treated with water, insulin and Tixosil particles at low or high doses, alone or coated with dried BfT. Animals were periodically weighed and monitored for water and food intake; urinary volume, glucose, urea and protein; blood glucose, serum lipids, liver toxicity markers transaminase and phosphatase and masses of adipose tissue and skeletal muscle. Insulin treatment gave best rat growth and lowest values for all other markers. No other treatment affected any diabetic marker, but the enzyme activities were changed by diabetes and BfT. Thus, BfT toxicity could arise from secondary products of plant constituents or Tixosil interaction. Therefore, BfT prepared in the spouted bed as described, is unsuitable for treatment of diabetes, which implies that the method of preparation of any medicine is critical for its efficacy and toxicity.

Protection against tuberculosis by a single intranasal administration of DNA-hsp65 vaccine complexed with cationic liposomes

Background: The greatest challenges in vaccine development include optimization of DNA vaccines for use in humans, creation of effective single-dose vaccines, development of delivery systems that do not involve live viruses, and the identification of effective new adjuvants. Herein, we describe a novel, simple technique for efficiently vaccinating mice against tuberculosis (TB). Our technique consists of a single-dose, genetic vaccine formulation of DNA-hsp65 complexed with cationic liposomes and administered intranasally. Results: We developed a novel and non-toxic formulation of cationic liposomes, in which the DNA-hsp65 vaccine was entrapped (ENTR-hsp65) or complexed (COMP-hsp65), and used to immunize mice by intramuscular or intranasal routes. Although both liposome formulations induced a typical Th1 pattern of immune response, the intramuscular route of delivery did not reduce the number of bacilli. However, a single intranasal immunization with COMP-hsp65, carrying as few as 25 mu g of plasmid DNA, leads to a remarkable reduction of the amount of bacilli in lungs. These effects were accompanied by increasing levels of IFN-gamma and lung parenchyma preservation, results similar to those found in mice vaccinated intramuscularly four times with naked DNA-hsp65 (total of 400 mu g). Conclusion: Our objective was to overcome the significant obstacles currently facing DNA vaccine development. Our results in the mouse TB model showed that a single intranasal dose of COMP-hsp65 elicited a cellular immune response that was as strong as that induced by four intramuscular doses of naked-DNA. This formulation allowed a 16-fold reduction in the amount of DNA administered. Moreover, we demonstrated that this vaccine is safe, biocompatible, stable, and easily manufactured at a low cost. We believe that this strategy can be applied to human vaccines to TB in a single dose or in prime-boost protocols, leading to a tremendous impact on the control of this infectious disease.

Structural basis of GC-1 selectivity for thyroid hormone receptor isoforms

Background: Thyroid receptors, TRa and TR beta, are involved in important physiological functions such as metabolism, cholesterol level and heart activities. Whereas metabolism increase and cholesterol level lowering could be achieved by TR beta isoform activation, TRa activation affects heart rates. Therefore, beta-selective thyromimetics have been developed as promising drug-candidates for treatment of obesity and elevated cholesterol level. GC-1 [ 3,5-dimethyl-4-(4'-hydroxy- 3'-isopropylbenzyl)-phenoxy acetic acid] has ability to lower LDL cholesterol with 600-to 1400-fold more potency and approximately two-to threefold more efficacy than atorvastatin (Lipitor(C)) in studies in rats, mice and monkeys. Results: To investigate GC-1 specificity, we solved crystal structures and performed molecular dynamics simulations of both isoforms complexed with GC-1. Crystal structures reveal that, in TRa Arg228 is observed in multiple conformations, an effect triggered by the differences in the interactions between GC-1 and Ser277 or the corresponding asparagine (Asn331) of TR beta. The corresponding Arg282 of TR beta is observed in only one single stable conformation, interacting effectively with the ligand. Molecular dynamics support this model: our simulations show that the multiple conformations can be observed for the Arg228 in TR alpha, in which the ligand interacts either strongly with the ligand or with the Ser277 residue. In contrast, a single stable Arg282 conformation is observed for TR beta, in which it strongly interacts with both GC-1 and the Asn331. Conclusion: Our analysis suggests that the key factors for GC-1 selectivity are the presence of an oxyacetic acid ester oxygen and the absence of the amino group relative to T(3). These results shed light into the beta-selectivity of GC-1 and may assist the development of new compounds with potential as drug candidates to the treatment of hypercholesterolemia and obesity.

Hypothalamic-pituitary axis and peripheral tissue responses to TRH stimulation and Liothyronine suppression tests in normal subjects evaluated by current methods

Objective: To reevaluate the responses of thyrotropin-releasing hormone ( TRH) stimulation test in baseline condition as well as after the administration of graded supraphysiological doses of liothyronine ( L- T-3) in normal subjects. Design: To assess various parameters related to the hypothalamic-pituitary axis and peripheral tissue responses to L- T-3 in 22 normal individuals ( median age: 30.5 years). Subjects were submitted to an intravenous TRH test at baseline condition and also to the oral administration of sequential and graded doses of L- T-3 ( 50, 100, and 200 mu g/day), each given over 3 days, at an outpatient clinic. Blood samples were obtained for thyrotropin (TSH) and prolactin (PRL) at basal and then 15, 30, and 60 minutes after the TRH injection. Effects of L- T3 administration on cholesterol, creatine kinase, retinol, ferritin, and sex hormone-binding globulin ( SHBG) were also measured at basal and after the oral administration of L- T-3. Main outcome: TRH administration resulted in an increase of 4-to 14-fold rise in serum TSH ( 8.3 +/- 2.5-fold), and in a slight rise in serum PRL concentrations ( 3.8 +/- 1.5-fold). Administration of graded doses of triiodothyronine ( T-3) resulted in a dose-dependent suppression of TSH and PRL. Basal thyroxine- binding globulin (TBG) and cholesterol levels decreased, and ferritin and SHBG increased after L- T-3 administration, while creatine kinase and retinol did not change throughout the study. There was a positive correlation between basal TSH and TSH peak response to TRH at basal condition and after each sequential L- T-3 doses. On the other hand, TSH peak response to the TRH test did not predict cholesterol, TBG, ferritin, or SHBG values. Conclusion: Using the current methods on hormone and biochemical analysis, we standardized the response of many parameters to TRH stimulation test after sequential and graded T-3 suppression test in normal subjects. Our data suggest that the evaluation of the responses of the hypothalamus-pituitary axis to TRH test as well as the impact of L- T-3 on peripheral tissues were not modified by the current methods.

Prognostic value of TP53 Pro47Ser and Arg72Pro single nucleotide polymorphisms and the susceptibility to gliomas in individuals from Southeast Brazil

The TP53 tumor suppressor gene codifies a protein responsible for preventing cells with genetic damage from growing and dividing by blocking cell growth or apoptosis pathways. A common single nucleotide polymorphism (SNP) in TP53 codon 72 (Arg72Pro) induces a 15-fold decrease of apoptosis-inducing ability and has been associated with susceptibility to human cancers. Recently, another TP53 SNP at codon 47 (Pro47Ser) was reported to have a low apoptosis-inducing ability; however, there are no association studies between this SNP and cancer. Aiming to study the role of TP53 Pro47Ser and Arg72Pro on glioma susceptibility and oncologic prognosis of patients, we investigated the genotype distribution of these SNPs in 94 gliomas (81 astrocytomas, 8 ependymomas and 5 oligodendrogliomas) and in 100 healthy subjects by the polymerase chain reaction-restriction fragment length polymorphism approach. Chi-square and Fisher exact test comparisons for genotype distributions and allele frequencies did not reveal any significant difference between patients and control groups. Overall and disease-free survivals were calculated by the Kaplan-Meier method, and the log-rank test was used for comparisons, but no significant statistical difference was observed between the two groups. Our data suggest that TP53 Pro47Ser and Arg72Pro SNPs are not involved either in susceptibility to developing gliomas or in patient survival, at least in the Brazilian population.