Activities report - Basic and Applied Sciences Branch, Division of Water Supply and Pollution Control

Report covers fiscal year

Harmine and imipramine promote antioxidant activities in prefrontal cortex and hippocampus

A growing body of evidence has suggested that reactive oxygen species (ROS) may play an important role in the physiopathology of depression. Evidence has pointed to the beta-carboline harmine as a potential therapeutic target for the treatment of depression. The present study we evaluated the effects of acute and chronic administration of harmine (5, 10 and 15 mg/kg) and imipramine (10, 20 and 30 mg/kg) or saline in lipid and protein oxidation levels and superoxide dismutase (SOD) and catalase (CAT) activities in rat prefrontal cortex and hippocampus. Acute and chronic treatments with imipramine and harmine reduced lipid and protein oxidation, compared to control group in prefrontal cortex and hippocampus. The SOD and CAT activities increased with acute and chronic treatments with imipramine and harmine, compared to control group in prefrontal cortex and hippocampus. In conclusion, our results indicate positive effects of imipramine antidepressant and beta-carboline harmine of oxidative stress parameters, increasing SOD and CAT activities and decreasing lipid and protein oxidation.

Detection of polymorphisms of the mtDNA control region of Caretta caretta (Testudines: Cheloniidae) by PCR-SSCP

Marine turtles are increasingly being threatened worldwide by anthropogenic activities. Better understanding of their life cycle, behavior and population structure is imperative for the design of adequate conservation strategies. The mtDNA control region is a fast-evolving matrilineal marker that has been employed in the study of marine turtle populations. We developed and tested a simple molecular tracing system for Caretta caretta mtDNA haplotypes by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Using this technique, we were able to distinguish the SSCP patterns of 18 individuals of the haplotypes CC-A4, CC-A24 and CCxLO, which are commonly found in turtles sampled on the Brazilian coast. When we analyzed 15 turtles with previously unknown sequences, we detected two other haplotypes, in addition to the other four. Based on DNA sequencing, they were identified as the CC-A17 and CC-A1 haplotypes. Further analyses were made with the sea turtles, Chelonia mydas (N = 8), Lepidochelys olivacea (N = 3) and Eretmochelys imbricata (N = 1), demonstrating that the PCR-SSCP technique is able to distinguish intra-and interspecific variation in the family Cheloniidae. We found that this technique can be useful for identifying sea turtle mtDNA haplotypes, reducing the need for sequencing.

In vitro and in vivo antileishmanial activities of a Brazilian green propolis extract

The in vitro antileishmanial activity of Brazilian green propolis hydroalcoholic extract (BPE) were carried out on Leishmania (Viannia) braziliensis against both promastigote (doses ranging from 1 to 750 mu g mL(-1)) and amastigote (10, 100, and 250 mu g mL(-1)) assays in comparison with the positive (amphotericin B) and negative (dimethyl sulfoxide at 1% in physiologic solution) control groups. BPE displayed in vitro antileishmanial activities against promastigote forms of the parasite (p<0.05). However, it was inactive against its amastigote ones. In the in vitro cytotoxicity assay against Vero cells, BPE showed no cytotoxicity in the maximum doses tested. The high-performance liquid chromatography analysis allowed the identification of caffeic acid, p-coumaric acid, aromadendrine-4`-methyl-ether, 3-prenyl-p-coumaric acid (drupanin), and 3,5-diprenil-p-cumarico acid (artepillin C) as major compounds of BPE. In the in vivo assay, using a Balb/C lineage of Mus musculus male mice, groups of ten animals each were treated (1.5 mg kg day(-1)) with BPE orally (group 1), BPE topically (group 2), BPE orally and topically (group 3), and glucantime (group 4), using NaCl 0.9% (group 5) as the negative control group. Groups 1, 2, and 3 displayed a decrease on lesion development, after 90 days of treatment, by 78.6%, 84.3%, and 90.0%, respectively, while the glucantime-treated group showed 57.7% of decrease, all in comparison with the negative control group. It is the first time that the in vivo antileishmanial activity has been reported for Brazilian green propolis.

Characterization and in vitro activities of cell-free antigens from Histoplasma capsulatum-loaded biodegradable microspheres

In the last decades, the incidence of histoplasmosis, a pulmonary fungal disease caused by Histoplasma capsulatum, has increased worldwide. In this context, vaccines for the prevention of this infection or therapies are necessary. Cell-free antigens (CFAgs) from H. capsulatum when administered for murine immunization purposes are able to confer protection and control of the infection, since they activate cellular immunity. However the most of vaccination procedures need several anti, gens administrations and immunoadjuvants, which are not approved for use in humans. The aim of this study was to develop and characterize a vaccination approach using biodegradable PLGA microspheres (MS) that could allow the controlled and/or sustained release of the encapsulated antigens from H. capsulatum. CFAgs-loaded MS presented a size less than 10 mu m, were marked engulfed by bone marrow-derived macrophages (BMDM phi) and induced the nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) production by these cells. Our data show that CFAgs-loaded MS induce cell activation, suggesting an immunostimulant effect to be further investigated during immunization procedures. CFAgs-loaded MS present potential to be used as vaccine in order to confer protection against H. capsulatum infection. (C) 2009 Elsevier B.V. All rights reserved.

Determination of Kininogens Levels and Kallikrein/Kininase II Activities in Patients with Thromboangiitis Obliterans

Some components of the kinin system such as plasma kallikrein levels, the activities of tissue kallikrein (including saliva) and kininase II and the concentrations of kininogen fractions (low-molecular weight/LKg and high-molecular weight/HKg) were evaluated in the plasma of patients with thromboangiitis obliterans (TAO) presenting clinical symptoms of the condition. Twenty TAO were diagnosed by means of the traditional Shionoya and Olin criteria and later classified into non-smokers (n = 11) and active smokers (n = 9). Fifty-three normal, non-smoking/smoking individuals (control) were also studied. Kininogen levels were determined by ELISA; the activities of kallikreins and kininase II were determined using selective substrates. The levels of enzymes (kallikreins and kininase II) and protein (kininogens) were significantly higher in patients with TAO who were active smokers compared to the control groups (no matter whether control individuals were active smokers or non-smokers, P < 0.001 for all comparisons). Interestingly, regardless of the time of disease onset, a significant increase in the levels of these components of the kinin system was also observed in patients when TAO active smokers were compared with TAO ex-smokers (P < 0.01 for all analysed parameters). Activation of the kinin system in patients with TAO may indicate the involvement of vasodilatation in an attempt to control vascular changes, thereby favouring the deposition of immune complexes at the vascular level because of nicotine stimulation. Moreover, our results corroborate the idea that TAO can be an autoimmune disorder with specific mechanisms.

Increased activities of cardiac matrix metalloproteinases matrix metalloproteinase (MMP)-2 and MMP-9 are associated with mortality during the acute phase of experimental Trypanosoma cruzi infection

The strong inflammatory reaction that occurs in the heart during the acute phase of Trypanosoma cruzi infection is modulated by cytokines and chemokines produced by leukocytes and cardiomyocytes. Matrix metalloproteinases (MMPs) have recently emerged as modulators of cardiovascular inflammation. In the present study we investigated the role of MMP-2 and MMP-9 in T. cruzi-induced myocarditis, by use of immunohistochemical analysis, gelatin zymography, enzyme-linked immunosorbent assay, and real-time polymerase chain reaction to analyze the cardiac tissues of T. cruzi-infected C57BL/6 mice. Increased transcripts levels, immunoreactivity, and enzymatic activity for MMP-2 and MMP-9 were observed by day 14 after infection. Mice treated with an MMP inhibitor showed significantly decreased heart inflammation, delayed peak in parasitemia, and improved survival rates, compared with the control group. Reduced levels of cardiac tumor necrosis factor-alpha, interferon-gamma, serum nitrite, and serum nitrate were also observed in the treated group. These results suggest an important role for MMPs in the induction of T. cruzi-induced acute myocarditis.

Coupling Between Respiratory and Sympathetic Activities as a Novel Mechanism Underpinning Neurogenic Hypertension

Enhanced sympathetic outflow to the heart and resistance vessels greatly contributes to the onset and maintenance of neurogenic hypertension. There is a consensus that the development of hypertension (clinical and experimental) is associated with an impairment of sympathetic reflex control by arterial baroreceptors. More recently, chronic peripheral chemoreflex activation, as observed in obstructive sleep apnea, has been proposed as another important risk factor for hypertension. In this review, we present and discuss recent experimental evidence showing that changes in the respiratory pattern, elicited by chronic intermittent hypoxia, play a key role in increasing sympathetic activity and arterial pressure in rats. This concept parallels results observed in other models of neurogenic hypertension, such as spontaneously hypertensive rats and rats with angiotensin II-salt-induced hypertension, pointing out alterations in the central coupling of respiratory and sympathetic activities as a novel mechanism underlying the development of neurogenic hypertension.

Postural control and movement performance during upper limb tasks in children with developmental co-ordination disorder (DCD)

At least 6% of primary school aged children present with DCD, where co-ordination is substantially below the normal range for the child’s age and intelligence. Motor skill difficulties negatively affect academic achievement, recreation and activities of daily living. Poor upper-limb co-ordination is a common difficulty for children with DCD. A possible cause of this problem is deviant muscle timing in proximal muscle groups, which results in poor postural and movement control. While studies have been published investigating postural control in response to external perturbations, detail about postural muscle activity during voluntary movement is limited even in children with normal motor development. No studies have investigated the relationship between muscle timing, resultant arm motion and upper-limb coordination deficits. Objectives: To investigate the relationship between functional difficulties with upper-limb motor skills and neuromuscular components of postural stability and coordination. Specifically, to investigate onset-timing of muscle activity, timing of arm movement, and resultant three-dimensional (3D) arm co-ordination during rapid, voluntary arm movement and to analyse differences arising due to the presence of DCD. This study is part of a larger research program investigating postural stability and control of upper limb movement in children. Design: A controlled, cross-sectional study of differences between children with and without DCD. Methods: This study included 50 children aged eight to 10 years (25 with DCD and 25 without DCD). Children participated in assessment of motor skills according to the Movement ABC Test and a laboratory study of rapid, voluntary arm movements. Parameters investigated included muscle activation timing of shoulder and trunk muscles (surface electromyography), arm movement timing (light sensor) and resultant 3D arm motion (Fastrak). Results: A MANOVA is being used to analyse between-group differences. Preliminary results indicate children with DCD demonstrate altered muscle timing during a rapid arm raise when compared with the control group of children. Conclusion: Differences in proximal muscle timing in children with DCD support the hypothesis that altered proximal muscle activity may contribute to poor proximal stability and consequently poor arm movement control. This has implications for clinical physiotherapy.

Chagas' disease and social security: a case-control study in an urban area, Goiás, Brazil

One hundred and twenty subjects with Chagas' cardiopathy and 120 non-infected subjects were randomly selected from first time claimants of sickness benefits in the National Institute of Social Security (INPS) in Goiás. Cases of Chagas' cardiopathy were defined based on serological test, history of residence in an endemic area and, clinical and/or electrocardiogram (ECG) alterations suggestive of Chagas' cardiomyopathy. Controls were defined as subjects with at least two negative serological tests. Case and controls were compared in the analysis for age, sex, place of birth, migration history, socio-economic level, occupation, physical exertion at work, age at affiliation and years of contribution to the social security scheme, clinical course of their disease and ECG abnormalities. Chagas' disease patients were younger than other subjects and predominantly of rural origin. Non-infected subjects presented a better socio-economic level, were performing more skilled activities and had less changes of job than cases. No important difference was observed in relation to age at affiliation to INPS. About 60% of cases have claimed for benefits within the first four years of contribution while among controls this proportion was 38.5%. Cases were involved, proportionally more than controls, in "heavy" activities. A risk of 2.3 (95%CL 1.5 - 4.6) and 1.8 (95%CL 1.2- 3.5) was obtained comparing respectively "heavy" and "moderate" physical activity against "light". A relative risk of 8.5 (95%CL 4.9 - 14.8) associated with the presence of cardiopathy was estimated comparing the initial sample of seropositive subjects and controls. A high relative risk was observed in relation to right bundle branch block (RR = 37.1 95%CL = 8.8 - 155.6) and left anterior hemiblock (RR = 4.4, 95%CL = 2.1 - 9.1).

Physical activity during pregnancy and fetal outcomes: a case-control study

OBJECTIVE: To investigate the relationship between physical activity during the second trimester pregnancy and low birth weight, preterm birth, and intrauterine growth restriction. METHODS: Case-control study including 273 low birth weight newborns and 546 controls carried out in the city of São Paulo, Southeastern Brazil, in 2005. Low birth weight cases were grouped into two subsamples: preterm birth (n=117) and intrauterine growth restriction (n=134), with their related controls. Information was collected by means of interviews with mothers shortly after birth and transcription of medical records. Data were analyzed using conditional multiple and hierarchical logistic regression. RESULTS: Light physical activity for over 7 hours per day was shown to be protective against low birth weight (adjusted OR=0.61; 95% CI 0.39-0.94) with a dose-response relationship (p-value for trend=0.026). A similar trend was found for intrauterine growth restriction (adjusted OR=0.51; 95% CI 0.26-0.97). Homemaking activities were associated as a protective factor for both low birth weight and preterm birth (p-value for trend=0.013 and 0.035, respectively). Leisure-time walking was found to be protective against preterm birth. CONCLUSIONS: Mild physical activity during the second trimester of pregnancy such as walking has an independent protective effect on low birth weight, preterm birth, and intrauterine growth restriction.

Autonomic workflow activities: the award framework

Workflows have been successfully applied to express the decomposition of complex scientific applications. This has motivated many initiatives that have been developing scientific workflow tools. However the existing tools still lack adequate support to important aspects namely, decoupling the enactment engine from workflow tasks specification, decentralizing the control of workflow activities, and allowing their tasks to run autonomous in distributed infrastructures, for instance on Clouds. Furthermore many workflow tools only support the execution of Direct Acyclic Graphs (DAG) without the concept of iterations, where activities are executed millions of iterations during long periods of time and supporting dynamic workflow reconfigurations after certain iteration. We present the AWARD (Autonomic Workflow Activities Reconfigurable and Dynamic) model of computation, based on the Process Networks model, where the workflow activities (AWA) are autonomic processes with independent control that can run in parallel on distributed infrastructures, e. g. on Clouds. Each AWA executes a Task developed as a Java class that implements a generic interface allowing end-users to code their applications without concerns for low-level details. The data-driven coordination of AWA interactions is based on a shared tuple space that also enables support to dynamic workflow reconfiguration and monitoring of the execution of workflows. We describe how AWARD supports dynamic reconfiguration and discuss typical workflow reconfiguration scenarios. For evaluation we describe experimental results of AWARD workflow executions in several application scenarios, mapped to a small dedicated cluster and the Amazon (Elastic Computing EC2) Cloud.

Autonomic Activities in the Execution of Scientific Workflows: Evaluation of the AWARD Framework

Workflows have been successfully applied to express the decomposition of complex scientific applications. However the existing tools still lack adequate support to important aspects namely, decoupling the enactment engine from tasks specification, decentralizing the control of workflow activities allowing their tasks to run in distributed infrastructures, and supporting dynamic workflow reconfigurations. We present the AWARD (Autonomic Workflow Activities Reconfigurable and Dynamic) model of computation, based on Process Networks, where the workflow activities (AWA) are autonomic processes with independent control that can run in parallel on distributed infrastructures. Each AWA executes a task developed as a Java class with a generic interface allowing end-users to code their applications without low-level details. The data-driven coordination of AWA interactions is based on a shared tuple space that also enables dynamic workflow reconfiguration. For evaluation we describe experimental results of AWARD workflow executions in several application scenarios, mapped to the Amazon (Elastic Computing EC2) Cloud.

A municipal level approach to the management of schistosomiasis control in Peri-Peri, MG, Brazil

A schistosomiasis control program was implemented between 1974/87 in Peri-Peri,. MG (622 inhabitants). Molluscicide (niclosamide) was applied at three monthly intervals in water sources with Biomphalaria glabrata, and individuals eliminating Schistosoma mansoni eggs in the feces were treated annually with oxamniquine. From 1974 to 1983 the control measures were undertaken by staff of the "René Rachou" Research Center FIOCRUZ (CPqRR), and from 1984 to 1987 these measures were included in the Capim Branco basic health network activities. During both periods, the prevalence, incidence, intensity of infection and hepatosplenic form as well as the number of infected snails decreased significantly. The prevalence decreased from 43.5 to 4.4%, the incidence from 19.0 to 2.9%, the overall intensity of S. mansoni from 281 to 87 and of the hepatosplenic form from 5.9 to 0.0%. The results obtained suggest that the municipal management of control measures was as effective as the vertical program conducted by CPqRR staff.