Cultivating global antimicrobial stewardship: linguistic and cultural validation of the Australian National Antimicrobial Prescribing Survey appropriateness assessment definitions for Portugal.

BACKGROUND: Antimicrobial resistance is driven by inappropriate antimicrobial prescribing. The National Antimicrobial Prescribing Survey (NAPS) is an Australian-developed auditing platform to assist in the assessments of antimicrobial quality by antimicrobial stewardship programmes using consensus-based definitions. The NAPS has demonstrated to be transferable to other countries. Its adaptation to Portugal could improve knowledge about the quality of antimicrobial prescribing in the country. OBJECTIVES: To translate, culturally adapt, and validate the Australian Hospital NAPS appropriateness assessment definitions of antimicrobial prescribing for Portugal. METHODS: International recommendations on translation and adaptation of instruments were followed. Two panels of experts participated in the process, using Zoom® for discussions and interviews, and Google Forms® for assessing vignettes. A native English-speaking person proficient in Portuguese conducted the back-translation. SPSS v.28 and Excel® were used for validity calculation. RESULTS: The Portuguese version was well accepted, its implementation being perceived as desirable and feasible by the experts. Validation process showed a Fleiss' κ score of 0.483 (95% CI, 0.415-0.551, P < 0.005) for appropriateness, and an average agreement with the Australian NAPS team of 0.8 and 0.9, respectively, for appropriateness and reasons for inappropriateness. CONCLUSIONS: The Portuguese version of the Australian Hospital NAPS appropriateness assessment definitions of antimicrobial prescribing, the first to be translated from English, was deemed non-inferior to the original, was well accepted, considered to be desirable and feasible, and could inspire other countries, particularly other Portuguese-speaking countries, to adapt and validate them in their own contexts, reinforcing the possibility of transferring NAPS use beyond Australia.

Resistance determinants of emerging pathogens isolated from an intensive care unit as a parameter of population health conditions of the Legal Amazon microregion.

Bacteria responsible for causing infections are common in hospital environments, water, soil, and food products. The infection risk is intensified by the absence of public sanitation, poor quality of life, and food scarcity. These external factors promote the dissemination of pathogens by direct contamination or biofilm formation. In this work, we identified bacterial isolates obtained from intensive care units in the southern region of Tocantins, Brazil. We compared matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques and 16S ribosomal ribonucleic acid (rRNA) molecular analysis; we also performed phenotypic characterization. Fifty-six isolates characterized using morphotinctorial tests were classified as gram-positive (80.4%; n = 45) and gram-negative (19.6%; n = 11) and were resistant to several antibiotic classes; notably, we identified the blaOXA-23 resistance gene in the ILH10 isolate. Microbial identification using MALDI-TOF MS resulted in the identification of Sphingomonas paucimobilis and Bacillus circulans. 16S rRNA sequencing revealed four isolates belonging to the genera Bacillus and Acinetobacter. The similarity was superior to 99% for Acinetobacter schindleri in the Basic Local Alignment Search Tool (BLAST), grouped in the clade superior to 90%. Several strains isolated from intensive care units (ICU) were resistant to various antibiotic classes. These techniques allowed for the identification of several microorganisms of importance in public health, enabling improvements in human infection control and proving the quality of inputs, food, and water.

Developing a social prescribing local system in a European Mediterranean country: a feasibility study to promote active and healthy aging.

BACKGROUND: Social Prescribing (SP) is an innovative strategy to respond to the non-clinical health needs of the population. A Social Prescribing Local System (SPLS) can be defined as a set of joined community, health, and social organizations to foster SP-oriented activities. This study aimed to develop and assess the feasibility of an SPLS implemented in a Mediterranean country, to promote health and wellbeing and contribute to active and healthy aging. METHODS: A mixed-methods approach was followed, including three sequential components: 1) Cross-sectional online survey targeting health professionals (HP) working in a primary health care cluster, Portugal's southern region; 2) Pilot study implementing an on-the-job training program for HP, designed to meet identified training needs in the survey; 3) Focus group (FG) with the HP who participated in the pilot study, two individual interviews, with an elderly patient and a community provider for assessing the satisfaction with the pilot test. RESULTS: Sixty-five HP completed the survey; of these, 13 completed the theoretical part of the on-the-job training program; and six (out of these 13) completed the full program. Five HP participated in the FG, one patient and one community provider were interviewed. The surveyed HP perceived as facilitators to implement SP: an automatic system of notifications to prompt the use of SP, contribute to patient satisfaction, human and community resources' stability. The survey also highlighted barriers to SP implementation: length of appointments, shortage of human resources, data records confidentiality, low patient adherence rates, bureaucratic issues, time constraints, and financial costs. Participants were satisfied with the training. Identified SPLS implementation benefits were grouped into four dimensions (from the qualitative approach): gains for patients' health and wellbeing, support for the health services, sustainability of the community resources, and HP' professional satisfaction. CONCLUSIONS: Our study took the first steps towards the implementation of an SPLS. Findings reinforce that training HP in SP and on-the-job training seems feasible. This approach was well received and appears to represent a suitable and sustainable strategy. It can promote professional satisfaction, support health services, contribute to the stability of community resources, improve health and promote active and healthy aging.

Person-specific calibration of a partial body counter used for individualised Am241skull measurements.

Incorporation of bone seeking alpha-emitting radionuclides such as241Am are of special concern, due to the potential of alpha particles to damage the extremely radiation-sensitive bone marrow. In the case of an internal contamination with241Am, directin vivomeasurements using Gamma-detectors are typically used to quantify the incorporated activity. Such detectors need to be calibrated with an anatomical phantom, for example of the skull, of known241Am activity that reproduces the anatomy of the measured individual as closely as possible. Any difference in anatomy and material composition between phantom and individual will bias the estimation of the incorporated activity. Consequently, in this work the impact of the most important anatomical parameters on detection efficiency of one of the germanium detectors of the Helmholtz Center Munich (HMGU) partial body counter were systematically studied. For that a detailed model of the germanium detector was implemented in the Monte Carlo codes GEANT4 and MCNPX. To simulate the detector efficiency, various skull voxel phantoms were used. By changing the phantom dimensions and geometry the impact of parameters such as shape and size of the skull, thickness of tissue covering the skull bone, distribution of241Am across the scull and within the skull bone matrix, on the detector efficiency was studied. Approaches to correct for these parameters were specifically developed for three physical skull phantoms for which Voxel phantoms were available: Case 102 USTUR phantom, Max-06 phantom, BfS phantom. Based on the impact of each parameter, correction factors for an 'individual-specific' calibration were calculated and applied to a real241Am contamination case reported in 2014. It was found that the incorporated241Am activity measured with the HMGU partial body counter was about twice as large as that estimated when using the BfS skull phantom without applying any correction factor for person-specific parameters. It is concluded that the approach developed in the present study should in the future be applied routinely for skull phantom measurements, because it allows for a considerably improved reconstruction of incorporated241Am using partial body counters.

Violacein antimicrobial activity on Staphylococcus epidermidis biofilm.

The aim of this study was to evaluate the antimicrobial potential of violacein (VIO) on Staphylococcus epidermidis biofilm. The minimum biofilm inhibition concentration (MBIC) and minimum biofilm eradication concentration (MBEC) were determined, as well as the effect of VIO exposure time on microbial viability in mature biofilm. Violacein showed good antibiofilm action, inhibiting biofilm formation and eradicating mature biofilm of S. epidermidis at concentrations of 20 µg.mL-1 and 160 µg.mL-1, respectively. At concentrations equal to MBEC and 2x MBEC, the biofilm was eradicated in 3 h and 2h30min of incubation, respectively.When evaluating VIO modulating effect on the action of clinically-used drugs (vancomycin, cefepime, ciprofloxacin and meropenem), especial synergism was observed in the violacein-ciprofloxacin association, it can completely erradicated the mature biofilm at the concentration of 1/2xMBEC and 1/4xMBEC, respectively. VIO shows good antimicrobial action on S. epidermidis biofilm and has the potential to synergistically modulate the activity of clinically-used antimicrobials.

Antichagasic effect of violacein from Chromobacterium violaceum.

AIMS: Violacein (VIO), a bacterial pigment produced by Chromobacterium violaceum, was examined to evaluate the antichagasic activity and its action mechanism against Trypanosoma cruzi Y strain. METHODS AND RESULTS: Violacein was tested against the epimastigote, trypomastigote and amastigote forms of T. cruzi Y strain (benznidazole-resistant strain). VIO inhibited all T. cruzi developmental forms, including amastigotes, which is implicated in the burden of infection in the chronic phase of Chagas disease (CD). VIO induced cell death in T. cruzi through apoptosis, as determined by flow cytometry analyses with specific molecular probes and morphological alterations, such as involvement of reactive oxygen species and changes in mitochondrial membrane potential and cell shrinkage. CONCLUSION: The results suggest antichagasic activity of VIO against T. cruzi Y strain with apoptotic involvement. SIGNIFICANCE AND IMPACT OF THE STUDY: The treatment of CD has limited efficacy and side effects that restrict patient tolerability and compliance. The VIO molecule could be used as a model for therapeutic alternatives for this disease.

Monte Carlo simulation of dose coefficients for a fish eye lens model exposed to monoenergetic electrons.

Vision is an important sense for the majority of the wildlife species, affecting their ability to find food and escape predation. Currently, no study on radiation induced cataract frequency on the fish eyes lens has been done. However, any thorough future study of this subject will require more accurate dose estimates for the fish eye lens than those currently available. For this purpose, the eye lens absorbed dose per unit fluence conversion coefficients for electron irradiation were calculated using the MCNPX Monte Carlo radiation transport code package. All results were validated against three different fish voxel models. The discrepancies between model results mainly originate from the different fish eye dimensions used in the different studies and in two of the cases the lack of a defined eye lens region. The dose conversion coefficients calculated in this work can be used to estimate the dose to the fish eye lens based on the activity concentration of the surrounding water. The model developed in this work has also demonstrated that the mathematical models still have several advantages over the voxel models.

Capacity to report on mortality attributable to chronic hepatitis B and C infections by Member States: An exercise to monitor progress towards viral hepatitis elimination.

Viral hepatitis is globally leading causes of death, and 96% of these are due to hepatitis B and C (HBV/HCV) late outcomes. The first Global Health Sector Strategy (GHSS) aims to reduce by 65% the mortality associated with HBV/HCV, and an indicator (C10) is proposed to monitor progress. Data on viral hepatitis and liver-related mortality are required, and different methods of estimation can be used, depending on availability and quality of sources. We aimed to understand the current situation and practicality of calculating C10, accessing available sources to estimate initial figure for Europe. We listed and compiled regional and national data sources reporting deaths from HCC, cirrhosis and chronic liver disease (CLD) and available estimates of attributable fraction. We critically appraised quality of data, highlighting gaps in current data and estimated mortality attributable to HBV and HCV, for 31 EU/EEA countries from 2010 to 2015. Mortality data are available for 30/31 countries. Quality varies but 60% of national sources report with specificity as required by WHO indicator. Attributable fraction is only available through the literature search. We estimated C10 for 87.6% country-years. Deaths attributable to HBV/HCV for this period and region were 292 600, while HCV deaths were three times higher. Incomplete data for 2015 prevented calculation of time trends. Regional sources are outdated for monitoring C10, but national sources are capable of reporting mortality data. Sources for attributable fraction are sparse, outdated and much needed. We recommend improvement of death registration allowing measuring this indicator. Studies measuring attributable fraction on national and subnational levels are crucial.

Equity-focused health impact assessment of Portuguese tobacco control legislation.

The World Health Organization recommend the Equity-Focused Health Impact Assessment (HIA) as a means to assess the impact of social and economic policies on the health of populations, and acknowledges their contribution to health inequality. We describe the application of the Equity-focused Impact Assessment methodology on the Portuguese law on Smoking Prevention and Tobacco Control (Law No. 37/2007). A rapid assessment was carried out to issue recommendations which could be incorporated into the law during a revision in 2014. Quantitative (consumption and health status indicators; equity analysis) and qualitative (Focus Group) approaches were taken to evaluate the impact of the law and formulate recommendations. Young people, men and women of low socioeconomic status, and pregnant women were identified as requiring specific and appropriate interventions to prevent smoking and support smoking cessation.

Violacein antimicrobial activity on Staphylococcus epidermidis and synergistic effect on commercially available antibiotics.

AIMS: The study aimed to assess whether violacein has antimicrobial activity on Staphylococcus epidermidis and synergistically modulates the action of commercially available antimicrobial drugs. METHODS AND RESULTS: Violacein showed excellent antimicrobial activity on biofilm-forming and nonbiofilm-forming S. epidermidis strains (ATCC 35984) (ATCC 12228), with bacteriostatic (MIC = 20 µg ml-1 and 10 µg ml-1 respectively) and bactericidal effects (MBC = 20 µg ml-1 for both strains), observed in short periods of exposure. The violacein bactericidal concentration led to S. epidermidis death after 2-3 h of exposure. Additionally, violacein synergistically modulated the activity of different antimicrobial classes on S. epidermidis ATCC 12228 (81·8%; n = 9) and on S. epidermidis ATCC 35984 (54·5%; n = 6), reducing the MIC of these antibiotics by up to 16-fold. CONCLUSION: Violacein shows excellent antimicrobial activity on S. epidermidis strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Violacein shows the potential for the development of a new drug for the treatment of infections caused by S. epidermidis.

Caries prevalence and treatment needs in young people in Portugal: the third national study.

OBJECTIVE: To evaluate caries prevalence and dental treatment needs in Portuguese children and teenagers, as well as fluorosis prevalence in 12-year-old children, in order to address public oral health program strategies. PARTICIPANTS: A representative stratified random cluster sample of 3,710 participants of 6, 12, and 18 years old was selected. A questionnaire was applied to determine sociodemographic data and some oral health determinants. The clinical examination was based on the ICDAS criteria, then adapted to DMFS index, and Dean's index for fluorosis. RESULTS: Caries prevalence at 6, 12, and 18 years old was 45.2%, 47.0%, and 67.6%, respectively. D5MFT scores were 1.18 (SD 0.06) and 2.51 (SD 0.10), respectively. Treatment needs at 12 and 18 years old were associated with 0.37 (SD 0.03) and 0.75 (SD 0.06) values in the "decayed" (D5) variable. SiC index at 12 years old was 2.68 (SD 1.68). Sealants were identified in 55% of 12-year-old children and the mean of sealants per individual was 3.61; also, moderate (2.2%) and severe (0.2%) levels of fluorosis were detected. CONCLUSIONS: The oral health situation in Portugal is favorable for young people, resulting in low treatment needs. The National Oral Health Promotion Program should be extended to include 18-year-olds.

Aqueous extract of Chrysobalanus icaco leaves, in lower doses, prevent fat gain in obese high-fat fed mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Due to the rise in obesity, the necessity for resources and treatments that could reduce the morbidity and mortality associated to this pandemia has emerged. The development of new anti-obesity drugs through herbal sources has been increasing in the past decades which are being used not only as medicine but also as food supplements. Previous studies with the aqueous extract of Chrysobalanus icaco L (AECI) have demonstrated activity on lowering blood glucose levels and body weight. AIM OF THE STUDY: Investigate C. icaco effects in overall adiposity and glycemic homeostasis. MATERIAL AND METHODS: C57BL/6J mice were randomly assigned to standard chow (SC) or high-fat diet (HFD) and treated with AECI in 0.35mg/mL or 0.7mg/mL concentrations ad libitum. Food intake, feed efficiency, metabolic efficiency, body, fat pads and gastrocnemius weight, adiposity index, serum lipids, fecal lipid excretion, locomotor activity in the open field test and insulin and glucose tolerance tests were analyzed and compared. The major components of the extract were demonstrated through HPLC and its antioxidant activity analyzed through DPPH and lipid peroxidation. RESULTS: The AECI in the 0.35mg/mL concentration did not affect food intake or body weight. However, it promoted lower adipose tissue gain, TG levels, and fecal lipid excretion, increased locomotor activity and lean mass weight, and normalized insulin sensitivity and glucose tolerance. Moreover, AECI showed the presence of myricetin 3-O-glucuronide, rutin, quercitrin and myricitrin and demonstrated high-antioxidant activity. CONCLUSIONS: AECI in lower concentrations can prevent fat storage or enhance fat utilization through the increase of locomotor activity. Also, this reinforces its ability to maintain glucose homeostasis through the normalization of insulin sensitivity and glucose tolerance despite the high-fat diet intake. These activities could be associated to the extract's polyphenol content.

EFFICIENCY STUDY OF A LEGe DETECTOR SYSTEM FOR THE ASSESSMENT OF 241Am IN SKULL AT CIEMAT WHOLE BODY COUNTER.

(241)Am incorporation due to an incident or chronic exposure causes an internal dose, which can be evaluated from the total activity of this isotope in the skeleton several months after the intake. For this purpose, it is necessary to perform in vivo measurements of this bone-seeker radionuclide in appropriate counting bone geometries with very low attenuation of surrounded tissue and to extrapolate to total activity in the skeleton (ICRP 89, Basic anatomical and physiological data for use in radiological protection: reference values. 2001. 265). The work here presented refers to direct measurements of americium in the Cohen skull phantom at the CIEMAT Whole Body Counter (WBC) using low-energy germanium (LEGe) detectors inside a shielding room. The main goal was to determinate the most adequate head counting geometry for the in vivo detection of americium in the bone. The calibration of the in vivo LEGe system was performed with four detectors with 2 cm of distance to Cohen phantom. Two geometries were measured, on junction of frontal to parietal bones and frontal bone. The efficiencies are very similar in both geometries, the preferred counting geometry is the most comfortable for the person, with the LEGe detectors in the highest part of the frontal bone, near the junction with the parietal bone, CIEMAT WBC participated in a skull intercomparison exercise organised by WG7 of EURADOS (European Radiation Dosimetry Group e.V.). Efficiencies using three different skull phantoms were obtained. Measurements were performed for different head counting positions, four of them in the plane of symmetry and others over the temporal bone. The detector was placed in parallel with the calibration phantom at a distance of 1 cm. The main gamma emission of (241)Am, 59.5 keV (36 %), was used for comparing efficiency values. The lower efficiency was obtained over the frontal and occipital bones. Measurement with one LEGe detector over the parietal bone is the most efficient. The activity of each skull phantom was calculated using CIEMAT head calibration. Results of the EURADOS intercomparison are presented here for discussion.

The effects of graphene oxide on green algae Raphidocelis subcapitata.

Graphene represents a new class of nanomaterials that has attracted great interest due to its unique electrical, thermal, and mechanical properties. Once disposed in the environment, graphene can interact with biological systems and is expected to exhibit toxicological effects. The ecotoxicity of graphene and its derivatives, viz.: graphene oxide (GO) depends on their physicochemical properties, including purity, diameter, length, surface charge, functionalization and aggregation state. In this study we evaluated the effects of graphene oxide (GO) on green algae Raphidocelis subcapitata. The algae were exposed to different concentrations of GO pre-equilibrated for 24h with oligotrophic freshwater medium (20ml) during incubation in a growth chamber under controlled conditions: 120µEm(-2)s(-1) illumination; 12:12h light dark cycle and constant temperature of 22±2°C. Algal growth was monitored daily for 96h by direct cell counting. Reactive oxygen species level (ROS), membrane damage (cell viability) and autofluorescence (chl-a fluorescence) were evaluated using fluorescent staining and further analyzed by flow cytometry. The toxic effects from GO, as observed in algal density and autofluorescence, started at concentrations from 20 and 10µgmL(-1), respectively. Such toxicity is probably the result of ROS generation and membrane damage (cell viability). The shading effect caused by GO agglomeration in culture medium may also contribute to reduce algal density. The results reported here provide knowledge regarding the GO toxicity on green algae, contributing to a better understanding of its environmental behavior and impacts.

Collaborative Brazilian Pediatric Renal Transplant Registry (CoBrazPed-RTx): A Report From 2004 to 2013.

BACKGROUND: The Collaborative Brazilian Pediatric Renal Transplant Registry started in 2004 as a multicenter initiative aiming to analyze, report, and share the results of pediatric kidney transplantation in Brazil. Data from all pediatric kidney transplants performed between January 2004 and December 2013 were recorded electronically and periodically updated. All patients under 18 years old from the participating centers were enrolled. Demographic data, etiology of chronic kidney disease, and patient and graft survival were analyzed. From a total of 2443 pediatric kidney transplants performed in Brazil during the study period, we report data from 1751 pediatric renal transplants performed in 13 centers enrolled in the collaborative study. Median age at transplantation was 12.4 years, and most of recipients were male (56%). The most common underlying renal etiologies were obstructive uropathy (31%) and glomerulopathy (26%). METHODS: According to donor source, 1155 (66%) of transplants were performed with deceased donors (DD). Initial immunosuppression consisted mainly of tacrolimus, mycophenolate, steroids, and induction therapy with anti-IL-2R antibodies. RESULTS: One-year graft survival (death-censored) was 93% and 90% (log rank test, P < .01), respectively, for living donor (LD) and DD. Graft losses (15%) were most frequently caused by vascular thrombosis, chronic allograft nephropathy, death with functioning kidney, acute rejection, and recurrent renal disease. Recipients of DD had 2.02 (95% confidence interval: 1.14-3.59) times the hazard of graft loss compared with those of LD (P = .015). Patient survival rates at 1 and 5 years were 98% and 97% for LD and 97% and 93% for DD, respectively. The mortality rate was 3.8%, mainly as the result of infection and cardiovascular disease. CONCLUSIONS: The results of this collaborative pediatric transplant study are comparable to international registries. Our effort has been able to maintain an exchange of information, both among the participating centers and with other international registries.

Immunological profile of periapical endodontic infections from HIV- and HIV+ patients.

AIM: To evaluate CD4(+) CD28(+) and CD8(+) T-cell genes and the gene expression of IFN-γ, TNF-α, IL-1-ß, IL-17A, IL-10, CCL-2/MCP-1, CCL-4, CCL-5 (RANTES), CXCR4, CCR5 and RANKL from cells in the periapical interstitial fluid from root canal infections in healthy patients (HIV-) and HIV-positive individuals (HIV+). METHODOLOGY: Subjects included 20 HIV- and 23 HIV+ patients referred to the School of Dentistry at the Universidade Federal de Minas Gerais (Belo Horizonte, MG, Brazil). Almost all HIV+ patients were undergoing highly active antiretroviral therapy (HAART). Clinical samples were taken from teeth with pulp necrosis, and no patients had acute periapical symptoms at the time of the appointments. After cleaning and drying, 3 paper points were introduced into the root canal, passing passively through the root apex (2 mm) into the periapical tissues for 1 min. The samples were collected immediately after root canal cleaning and 7 days later (restrained root canal bacterial load) to characterize those gene expressions using real-time PCR. RESULTS: Significantly higher levels of CD4(+) CD28(+) and CD8(+) T cells in teeth with restrained bacterial loads (second collection) compared with the first collection were observed in both HIV- and HIV+ samples. In HIV- patients, an increase in IL-10 and CXCR4 expression was demonstrated as well as a decrease in pro-inflammatory cytokines such as RANKL, IFN-γ, IL1-ß and CCL5. However, in HIV+ patients an increase in cytokines IFN-γ, IL-1-ß, TNF-α and IL-17A, and chemokines CCL-2, CXCR4 and CCR5 were observed. The chemokine CCL-5 was not detected in HIV+ individuals. CONCLUSIONS: These findings suggest that after reducing the root canal bacterial load in HIV- individuals an anti-inflammatory response is generated whilst in HIV+ patients a pro-inflammatory response is sustained in the periapical area.

Method for obtaining high-resolution proteomic analysis from pericarps of guarana.

Guarana has great agricultural potential and is largely used therapeutically and in the production of non-alcoholic energy drinks. Genomic and proteomic studies are crucial to identify proteins that play central roles in the maintenance and viability of fruits, as well as to identify proteins related to the main metabolic pathways. However, the success of any protein analysis starts with the protein extract preparation, which needs to offer an extract that is free of contaminants. This study aimed to evaluate different extraction methods to obtain high-quantity and high-quality extracts that are compatible with analysis by 2-dimensional electrophoresis and tandem mass spectrometry protein identification. Three different methods were tested: trichloroacetic acid (TCA)/acetone, sodium dodecyl sulfate (SDS)/phenol, and polyvinylpolypyrrolidone (PVPP)/SDS/phenol. The extract obtained from the TCA/acetone precipitation presented low solubility and contamination with lipids and carbohydrates. On the other hand, the quality of the extract gradually improved after using phenol and PVPP/phenol, enabling a yield up to 2 mg/g macerated tissues and the detection of 457 spots by 2-dimensional electrophoresis. The effectiveness of the procedure used was validated by identification of 10 randomly selected proteins by mass spectrometry. The procedure described here can be a starting point for applications using tissues of other organs of guarana or tissues of species that are similar to guarana.

Involvement of cerebral nervous system areas and cytokines on antihyperalgesic and anti-inflammatory activities of Kielmeyera rugosa Choisy (Calophyllaceae) in rodents.

Kielmeyera rugosa is a medicinal plant known in Northeastern Brazil as 'pau-santo', and it is used in the treatment of several tropical diseases such as malaria, schistosomiasis, and leishmaniasis. We evaluated antihyperalgesic and anti-inflammatory activities of methanol stem extract of K. rugosa (MEKR) in mice. The mechanical hyperalgesia induced by carrageenan and tumor necrosis factor-alpha (TNF-α), prostaglandin E2 , and dopamine were assessed. We also investigated the anti-inflammatory effect of MEKR on carrageenan-induced pleurisy and paw edema. Ninety minutes after the treatment, the animals were submitted to an imunofluorescence for Fos protein. MEKR (100, 200, and 400 mg/kg; p.o.) inhibited the development of mechanical hypernociception and edema. MEKR significantly decreased TNF-α and interleukin 1ß levels in pleural lavage and suppressed the recruitment of leukocytes. MEKR (1, 10, and 100 mg/mL) did not produce cytotoxicity, determined using the methyl-thiazolyl-tetrazolium assay in vitro. The locomotor activity was not affected. MEKR activated significantly the bulb olfactory, piriform cortex, and periaqueductal gray of the central nervous system. Our results provide first time evidence to propose that MEKR attenuates mechanical hyperalgesia and inflammation, in part, through an activation of central nervous system areas, mainly the periaqueductal gray and piriform cortex areas.