Prenatal toxoplasmosis diagnosis from amniotic fluid by PCR

Toxoplasmosis is one of the most common infections all over the world. Most cases are asymptomatic, except in immunosuppressed individuals and fetuses, which can be seriously damaged. Prenatal diagnosis should be made as soon as possible since treatment of the mother can minimize fetal sequelae. Our aim in this study was to test the polymerase chain reaction technique (PCR) in 86 samples of amniotic fluid from women who seroconverted during pregnancy. DNA was amplified using external primers and, in a second step, internal primers, in a nested PCR system. Samples were also inoculated into mice and the newborn were evaluated by T. gondii serology, skull x-ray, transfontanel ultrasound, fundoscopic examination, lumbar puncture and clinical examination. PCR was positive in seven cases and negative in 79. Among PCR-positive cases, two were negative by inoculation into mice and by clinical evaluation; among PCR-negative ones, three had clinical evidence of toxoplasmosis and one was positive after inoculation into mice. PCR showed values of sensitivity = 62.5% and specificity = 97.4%; the values of inoculation into mice where 42.9% and 100%, respectively. Although PCR should not be used alone for prenatal diagnosis of congenital toxoplasmosis, it is a promising method and deserves more studies to improve its efficacy.

Time-variations of equivalent water heights'from Grace Mission and in-situ river stages in the Amazon basin

Gravity Recovery and Climate Experiment (GRACE) mission is dedicated to measuring temporal variations of the Earth's gravity field. In this study, the Stokes coefficients made available by Groupe de Recherche en Géodésie Spatiale (GRGS) at a 10-day interval were converted into equivalent water height (EWH) for a ~4-year period in the Amazon basin (from July-2002 to May-2006). The seasonal amplitudes of EWH signal are the largest on the surface of Earth and reach ~ 1250mm at that basin's center. Error budget represents ~130 mm of EWH, including formal errors on Stokes coefficient, leakage errors (12 ~ 21 mm) and spectrum truncation (10 ~ 15 mm). Comparison between in situ river level time series measured at 233 ground-based hydrometric stations (HS) in the Amazon basin and vertically-integrated EWH derived from GRACE is carried out in this paper. Although EWH and HS measure different water bodies, in most of the cases a high correlation (up to ~80%) is detected between the HS series and EWH series at the same site. This correlation allows adjusting linear relationships between in situ and GRACE-based series for the major tributaries of the Amazon river. The regression coefficients decrease from up to down stream along the rivers reaching the theoretical value 1 at the Amazon's mouth in the Atlantic Ocean. The variation of the regression coefficients versus the distance from estuary is analysed for the largest rivers in the basin. In a second step, a classification of the proportionality between in situ and GRACE time-series is proposed.

Quantitative method of viral pollution determination for large volume of water using ferric hydroxide gel impregnated on the surface of glassfibre cartridge

Quantitative method of viral pollution determination for large volume of water using ferric hydroxide gel impregnated on the surface of glassfibre cartridge filter. The use of ferric hydroxide gel, impregnated on the surface of glassfibre cartridge filter enable us to recover 62.5% of virus (Poliomylitis type I, Lsc strain) exsogeneously added to 400 liters of tap-water. The virus concentrator system consists of four cartridge filters, in which the three first one are clarifiers, where the contaminants are removed physically, without significant virus loss at this stage. The last cartridge filter is impregnated with ferric hydroxide gel, where the virus is adsorbed. After the required volume of water has been processed, the last filter is removed from the system and the viruses are recovered from the gel, using 1 liter of glycine/NaOH buffer, at pH 11. Immediately the eluate is clarified through series of cellulose acetate membranes mounted in a 142mm Millipore filter. For the second step of virus concentration, HC1 1N is added slowly to the eluate to achieve pH 3.5-4. MgC1, is added to give a final concentration of 0.05M and the viruses are readsorbed on a 0.45 , porosity (HA) cellulose acetate membrane, mounted in a 90 mm Millipore filter. The viruses are recovered using the same eluent plus 10% of fetal calf serum, to a final volume of 3 ml. In this way, it was possible to concentrate virus from 400 liters of tap-water, into 1 liter in the first stage of virus concentration and just to 3 ml of final volume in a second step. The efficiency, simplicity and low operational cost, provded by the method, make it feasible to study viral pollution of recreational and tap-water sources.

Doenças tropicais negligenciadas: uma nova era de desafios e oportunidades

In an article recently published in Química Nova, entitled "Chemistry Without Borders" ("Química Sem Fronteiras") [Pinto, A. C.; Zucco, C.; Galembeck, F.; Andrade, J. B.; Vieira, P. C. Quim. Nova 2012, 35, 2092], the authors highlighted the important aspects of science and technology with special emphasis on the field of Chemistry and its contributions toward a more prosperous Brazil of future. As a second step in that direction, this article extends the discussion of a key issue for the country in the framework of the chemistry community through the so called position papers in strategic areas. This document is a part of the contribution of the Brazilian Chemical Society to the World Science Forum to be held in Rio de Janeiro in November 2013. In this context, the present paper provides a brief discussion on neglected tropical diseases (NTDs) with emphasis on the current challenges and opportunities towards the development and evolution of the field. NTDs leads to illness, long-term disability or death, and has severe social, economic and psychological consequences for millions of men, women, and children worldwide. In most cases, the available treatments are inadequate and extremely limited in terms of efficacy and safety, leading to an urgent demand for new drugs. In addition to the traditional challenges involved in any drug discovery process, it is widely recognized that there is an innovation gap and a lack of investment for research and development (R&D) in the area of NTDs. In the last few decades, methods toward combating, eradication, prevention, and treatment of NTDs have been repeatedly emphasized in the major international agendas. Developments in these strategies and alliances have continued to have an essential impact, particularly in the area of drug discovery, both in Brazil and globally and should be encouraged and supported. Several examples of international activities dedicated to the reduction of the devastating global impact of NTDs can be provided. Despite the beneficial developments in the past 30 years, NTDs continue to devastate poor communities in remote and vulnerable areas, in large part, due to market failures and public policies. Recent studies have shown that among 756 new drugs approved between 2000 and 2011, only four new chemical entities (NCEs) were identified for the treatment of malaria, while none were developed against NTDs or tuberculosis. Furthermore, only 1.4% of approximately 150,000 clinical trials were registered for neglected diseases, with a smaller number of trials for NCEs. Establishment and strengthening of global strategies involving the triad "government-academia-industry" is fundamental to the success in R&D of new drugs for NTDs. National and international public-private initiatives that aim to create, encourage, and invest in R&D projects have been implemented and therefore are of utmost importance to successfully integrate Brazil into this new paradigm. It is essential to lay the foundation for mechanisms that will intensify investments in infrastructure, training, and qualification of personnel with an ultimate strategic vision that foresees continuity. Our research group has made significant contributions to the development of this field with the goal of forging new frontiers while tackling both current and future challenges that include indispensable elements such as innovation and integration.

OBTENÇÃO DE COMPÓSITOS HÍBRIDOS BASEADOS EM HIDROGÉIS E CIMENTO PORTLAND

In this study, a novel hybrid composite based on biodegradable hydrogel and Portland cement with promising technological properties was reported. In the first step, a full 23 with central point factorial design was utilized to obtain the enhanced polyacrylamide-carboxymethylcellulose hydrogel compositions. A mathematical model was devised, indicating that the 3 main variables were significant and the AAm and MBAAm variables positively contributed to the mode and showing that the CMC variable had the opposite contribution. In the second step, these compositions were mixed with Portland cement to obtain the hybrid composites. The presence of cement improved the mechanical properties of polymeric matrices, and electronic microscopic micrographics revealed that the hydrogels were well adhered to the cement phase and no phase separation between hydrogel and cement was detected. Finally, using the energy dispersive X-ray technique, the elements Na, Mg, Al, Si, S, K, Ca and Fe were detected in the polymeric matrix, consistent with the hybrid composite formation.

Control of lettuce bottom rot by isolates of Trichoderma spp

Bottom rot, caused by Rhizoctonia solani AG 1-IB, is an important disease affecting lettuce in Brazil, where its biological control with Trichoderma was not developed yet. The present study was carried out with the aim of selecting Trichoderma isolates to be used in the control of lettuce bottom rot. Forty-six Trichoderma isolates, obtained with baits containing mycelia of the pathogen, were evaluated in experiments carried out in vitro and in vivo in a greenhouse in two steps. In the laboratory, the isolates were evaluated for their capabilities of parasitizing and producing toxic metabolic substances that could inhibit the pathogen mycelial growth. In the first step of the in vivo experiments, the number and the dry weight of lettuce seedlings of the cultivar White Boston were evaluated. In the second step, 12 isolates that were efficient in the first step and showed rapid growth and abundant sporulation in the laboratory were tested for their capability of controlling bottom rot in two repeated experiments, and had their species identified. The majority of the isolates of Trichoderma spp. (76%) showed high capacity for parasitism and 50% of them produced toxic metabolites capable of inhibiting 60-100% of R. solani AG1-IB mycelial growth. Twenty-four isolates increased the number and 23 isolates increased the dry weight of lettuce seedlings inoculated with the pathogen in the first step of the in vivo experiments.In both experiments of the second step, two isolates of T. virens, IBLF 04 and IBLF 50, reduced the severity of bottom rot and increased the number and the dry weight of lettuce seedlings inoculated with R. solani AG1-IB. These isolates had shown a high capacity for parasitism and production of toxic metabolic substances, indicating that the in vitro and in vivo steps employed in the present study were efficient in selecting antagonists to be used for the control of lettuce bottom rot.

Comparative analysis of different meat traceability systems using multiple criteria and a social network approach

The adoption of a proper traceability system is being incorporated into meat production practices as a method of gaining consumer confidence. The various partners operating in the chain of meat production can be considered a social network, and they have the common goal of generating a communication process that can ensure each characteristic of the product, including safety. This study aimed to select the most appropriate meat traceability system “from farm to fork” that could be applied to Brazilian beef and pork production for international trade. The research was done in three steps. The first used the analytical hierarchy process (AHP) for selecting the best on-farm livestock traceability. In the second step, the actors in the meat production chain were identified to build a framework and defined each role in the network. In the third step, the selection of the traceability system was done. Results indicated that with an electronic traceability system, it is possible to acquire better connections between the links in the chain and to provide the means for managing uncertainties by creating structures that facilitate information flow more efficiently.

Banda gástrica ajustável laparoscópica: alternativa técnica

In these paper we are presenting a technical alternative to laparoscopic adjustable gastric banding. From January 1999 to April 2000, 60 patients with mean body mass index (BMI) of 40,7 kg/m2 underwent laparoscopic adjustable gastric banding. The new technique is performed in two steps. In the first step, an isolation instrument (laparoscopic finger) is inserted through the lesser sac, next to the junction of diaphragmatic crura, including the lesser omentum in order to pull the band catheter. The second step separates the lesser omentum from the right side of the stomach.There was no mortality and the morbidity was 11,6% (1 slippage of the band and 6 trocar port seroma). The new technique was performed in all patients with no conversion to open procedure. We didn't have respiratory complications. This technical alternative is safe and easily performed, helping to prevent transoperative perforations.

Translation into Brazilian Portuguese, cultural adaptation and evaluation of the reliability of the Disabilities of the Arm, Shoulder and Hand Questionnaire

The objective of the present study was to translate, adapt and validate a Brazilian Portuguese version of the Disabilities of the Arm, Shoulder and Hand (DASH) Questionnaire. The study was carried out in two steps. The first was to translate the DASH into Portuguese and to perform cultural adaptation and the second involved the determination of the reliability and validity of the DASH for the Brazilian population. For this purpose, 65 rheumatoid arthritis patients of either sex (according to the classification criteria of the American College of Rheumatology), ranging in age from 18 to 60 years and presenting no other diseases involving the upper limbs, were interviewed. The patients were selected consecutively at the rheumatology outpatient clinic of UNIFESP. The following results were obtained: in the first step (translation and cultural adaptation), all patients answered the questions. In the second step, Spearman's correlation coefficients for interobserver evaluation ranged from 0.762 to 0.995, values considered to be highly reliable. In addition, intraclass correlation coefficients ranged from 0.97 to 0.99, also highly reliable values. Spearman's correlation coefficients and the intraclass correlation coefficients obtained during intra-observer evaluation ranged from 0.731 to 0.937 and from 0.90 to 0.96, respectively, being highly reliable values. The Ritchie Index showed a weak correlation with Brazilian DASH scores, while the visual analog scale of pain showed a good correlation with DASH score. We conclude that the Portuguese version of the DASH is a reliable instrument.

Asthma: where is it going?

Asthma is characterized by reversible airway obstruction, airway hyperresponsiveness, and airway inflammation. Although our understanding of its pathophysiological mechanisms continues to evolve, the relative contributions of airway hyperresponsiveness and inflammation are still debated. The first mechanism identified as important for asthma was bronchial hyperresponsiveness. In a second step, asthma was recognized also as an inflammatory disease, with chronic inflammation inducing structural changes or remodeling. However, persistence of airway dysfunction despite inflammatory control is observed in chronic severe asthma of both adults and children. More recently, a potential role for epithelial-mesenchymal communication or transition is emerging, with epithelial injury often resulting in a self-sustaining phenotype of wound repair modulation by activation/reactivation of the epithelial-mesenchymal trophic unit, suggesting that chronic asthma can be more than an inflammatory disease. It is noteworthy that the gene-environmental interactions critical for the development of a full asthma phenotype involve processes similar to those occurring in branching morphogenesis. In addition, a central role for airway smooth muscle in the pathogenesis of the disease has been explored, highlighting its secretory function as well as different intrinsic properties compared to normal subjects. These new concepts can potentially shed light on the mechanisms underlying some asthma phenotypes and improve our understanding of the disease in terms of the therapeutic strategies to be applied. How we understand asthma and its mechanisms along time will be the focus of this overview.

Single-step purification of crotapotin and crotactine from Crotalus durissus terrificus venom using preparative isoelectric focusing

We describe the isolation of crotoxin, a presynaptic B-neurotoxin, as well as its subunits B (crotactine) and A (crotapotin) from lyophilized Crotalus durissus terrificus venom by a single-step preparative isoelectric focusing procedure. From 98 mg of dried venom protein 20.1 mg of crotactine and 13.1 mg of crotapotin were recovered in the first step of focalization and 4.2 mg in a second run. These values correspond to 35.7% of the total venom protein applied. Crotactine separated in the 9.3-7.0 pH range (tubes 1-6) and crotapotin in the 1.8-2.8 pH range (tubes 15-19) and both were homogeneous by SDS-PAGE and N-terminal amino acid analysis. Crotactine, a 12-kDa protein, presented hemolytic and phospholipase A2 activity. Thus, using isoelectric focusing we simultaneously purified both toxins in high yields. This method can be used as an alternative for the purification and characterization of proteins from other snake venoms under conditions in which biological activity is retained

Sustainable Tourism and Eradication of Poverty (Step): impact assessment of a tourism development program in Brazil

This paper aims to evaluate the social impacts of the Tourism Development Program (Prodetur) in the northeastern town of Porto Seguro, Bahia, Brazil. The method used is based on the difference in difference technique applied to the 1991 and 2000 Census microdata. The results suggest social advances following from poverty relief based on income - where the benefits are distributed, generally, in a relatively equal manner between the native and migrant population. There is a relative deterioration in the sanitary situation, which consists of a very serious problem in the mid- and long-term, whose costs are mostly borne by the native population. Therefore, maintaining the natural capital is the main aspect that distances Porto Seguro’s tourism supply from the concept of sustainability. The article also relies on difference in difference estimators to assess the impacts of local public policies related to the sector.

Brazilian doctors' perspective on the second opinion strategy before a C-section

OBJECTIVE: To describe the opinion of doctors who participated in the Latin American Study on Cesarean section in Brazil regarding the second opinion strategy when faced with the decision of performing a C-section. METHODS: Seventy-two doctors from the hospitals where the study took place (where the second opinion was routinely sought) and 70 from the control group answered a pre-tested self-administered structured questionnaire. Descriptive tables were prepared based on the frequency of relevant variables on opinion of physicians regarding: effectiveness of the application of the second opinion strategy; on whether they would recommend implementation of this strategy and reasons for not recommending it in private institutions; feasibility of the strategy implementation and reasons for not considering this implementation feasible in private institutions. RESULTS: Half of the doctors from the intervention hospitals (50%) and about two thirds of those in the control group (65%) evaluated the second opinion as being or having the potential of being effective/very effective in their institutions. The great majority of those interviewed from both intervention and control hospitals considered this strategy feasible in public (87% and 95% respectively) but not in private hospitals (64% and 70% respectively), mainly because in the latter the doctors would not accept interference from a colleague in their decision-making process. CONCLUSION: Although the second opinion strategy was perceived as effective in reducing C-section rates, doctors did not regard it feasible outside the public health system in Brazil.

Towards a unified theory of health-disease: II. Holopathogenesis

This article presents a systematic framework for modeling several classes of illness-sickness-disease named as Holopathogenesis. Holopathogenesis is defined as processes of over-determination of diseases and related conditions taken as a whole, comprising selected facets of the complex object Health. First, a conceptual background of Holopathogenesis is presented as a series of significant interfaces (biomolecular-immunological, physiopathological-clinical, epidemiological-ecosocial). Second, propositions derived from Holopathogenesis are introduced in order to allow drawing the disease-illness-sickness complex as a hierarchical network of networks. Third, a formalization of intra- and inter-level correspondences, over-determination processes, effects and links of Holopathogenesis models is proposed. Finally, the Holopathogenesis frame is evaluated as a comprehensive theoretical pathology taken as a preliminary step towards a unified theory of health-disease.