ATPergic signaling disruption in human sepsis as a potential source of biomarkers for clinical use.

Sepsis is a life-threatening organ dysfunction caused by a dysregulated inflammatory response to infection. To date, there is no specific treatment established for sepsis. In the extracellular compartment, purines such as adenosine triphosphate (ATP) and adenosine play essential roles in the immune/inflammatory responses during sepsis and septic shock. The balance of extracellular levels among ATP and adenosine is intimately involved in the signals related to immune stimulation/immunosuppression balance. Specialized enzymes, including CD39, CD73, and adenosine deaminase (ADA), are responsible to metabolize ATP to adenosine which will further sensitize the P2 and P1 purinoceptors, respectively. Disruption of the purinergic pathway had been described in the sepsis pathophysiology. Although purinergic signaling has been suggested as a potential target for sepsis treatment, the majority of data available were obtained using pre-clinical approaches. We hypothesized that, as a reflection of deregulation on purinergic signaling, septic patients exhibit differential measurements of serum, neutrophils and monocytes purinergic pathway markers when compared to two types of controls (healthy and ward). It was observed that ATP and ADP serum levels were increased in septic patients, as well as the A2a mRNA expression in neutrophils and monocytes. Both ATPase/ADPase activities were increased during sepsis. Serum ATP and ADP levels, and both ATPase and ADPase activities were associated with the diagnosis of sepsis, representing potential biomarkers candidates. In conclusion, our results advance the translation of purinergic signaling from pre-clinical models into the clinical setting opening opportunities for so much needed new strategies for sepsis and septic shock diagnostics and treatment.

Ketoprofen-loaded rose hip oil nanocapsules attenuate chronic inflammatory response in a pre-clinical trial in mice.

This study aimed to develop nanocapsules containing ketoprofen using rose hip oil (Keto-NC) as oil core, and to evaluate their anti-inflammatory activity in acute and chronic ear edema models in mice. Physicochemical characterization, drug release, photostability and cytotoxicity assays were performed for the developed Keto-NC formulations and compared to ketoprofen-loaded nanocapsules using medium chain triglycerides as oil core (Keto-MCT-NC). Anti-inflammatory activity of orally delivered KP (Ketoprofen-free; 10 mg.kg-1) or Keto-NC (2.5; 5; 10 mg.kg-1) was assessed in mouse acute and chronic ear edema induced by croton oil (CO). Edema histological characteristics were determined by H&E stain, and redox parameters were analyzed in blood plasma and erythrocytes. Keto-MCT-NC and Keto-NC did not exhibit differences regarding physicochemical parameters, including size diameters, polydispersity index, pH, Ketoprofen content, and encapsulation efficiency. However, Keto-NC, which contains rose hip oil as lipid core, decreased drug photodegradation under UVC radiation when compared to Keto-MCT-NC. KP or Keto-NC were not cytotoxic to keratinocyte cultures and produced equal edema inhibition in the acute protocol. Conversely, in the chronic protocol, Keto-NC was more effective in reducing edema (~60-70% on 7-9th days of treatment) when compared to KP (~40% on 8-9th days of treatment). This result was confirmed by histological analysis, which indicated reduction of edema and inflammatory infiltrate. A sub-therapeutic dose of Keto-NC (5 mg.kg-1) significantly reduced edema when compared to control. Finally, KP and Keto-NC exhibited similar effects on redox parameters, suggesting that the advantages associated with Ketoprofen nanoencapsulation did not involve oxidative stress pathways. The results showed that Keto-NC was more efficient than KP in reducing chronic inflammation. These data may be important for the development of strategies aiming treatment of chronic inflammatory diseases with fewer adverse effects.

Ventilation distribution, pulmonary diffusion and peripheral muscle endurance as determinants of exercise intolerance in elderly patients with chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is a progressive and disabling disease that has been associated with aging. Several factors may potentially impair performance during exercise in elderly patients with COPD. This study was conducted to evaluate what characteristics related to lung function, peripheral muscle strength and endurance can predict the performance of elderly patients with COPD during cardiopulmonary exercise testing (CPET). Forty elderly patients with COPD underwent resting lung function tests, knee isokinetic dynamometry, and CPET. Three models were developed to explain the variability in peak oxygen uptake (VO(2) peak) after controlling for age as an independent confounder. The pulmonary function model showed the highest explained variance (65.6 %); in this model, ventilation distribution (p<0.001) and pulmonary diffusion (0.013) were found to be independent predictors. Finally, the models that included the muscle strength and endurance variables presented explained variances of 51 % and 57.4 %, respectively. In these models that involved muscular dysfunction, however, only the endurance variables were found to be independent predictors (p<0.05). In conclusion, ventilation distribution and pulmonary diffusion, but not the degree of airway obstruction, independently predict CPET performance in elderly patients with COPD. In addition, peripheral muscle endurance, but not strength, also predicts CPET performance in these subjects.

Periodontal disease detection in primary and mixed dentitions.

AIM: This was to compare the periodontal status of children with primary and mixed dentition at the time of their first consultation. METHODS: Children (200), aged 0-12 years (156 with mixed and 44 with primary dentition), were examined by assessing their simplified plaque index (PI) and simplified periodontal record (PSR). Statistical analysis (Chi-square test) was performed with appropriate software to find any significant associations between sex, type of dentition and PI with the PSR codes (0, 1, 2, 3 and 4). RESULTS: There was no statistically significant difference with regard to gender (p = 0.82). Generalised PI was associated more significantly with mixed dentition (p = 0.025 and higher PSR scores (p < 0.001). There was no significant relationship between sex and PSR codes (p = 0.82). CONCLUSION: Children presenting with a mixed dentition had worse PI values and PSR scores. It is important to perform periodontal examination in children to diagnose and prevent future periodontal disease and maintain their dentition as well as to identify any associated systemic conditions.

Outbreak of herpangina in the Brazilian Amazon in 2009 caused by Enterovirus B.

In October 2009, our laboratory was contacted by a Brazilian Public Health organization regarding a severe community outbreak of an acute exanthematic and febrile disease in the Brazilian Amazon that primarily affected children. A total of 44 patients with febrile disease were identified by the local public health system, 37 of whom were children between 1 and 9 years of age. Molecular virological and phylogenetic characterization revealed that enterovirus B was the etiological agent of this outbreak, which was characterized by a clinical presentation known as herpangina.

A 15-year follow-up study on schistosomiasis in a low-endemic area in Rio de Janeiro State, Brazil.

Fifteen years after our first investigation, a follow-up study was carried out with the purpose of assessing the evolution of schistosomiasis in the locality of Sabugo, Paracambi, state of Rio de Janeiro, Brazil, an area with low prevalence of the disease. The coprological techniques adopted were spontaneous sedimentation and Kato-Katz. Out of the 1356 individuals assessed, 13 (1%) were infected with Schistosoma mansoni. From those, 10 were males, 12 were over 15 years old, and at least 11 had been infected in Sabugo. All patients presented either the intestinal or the hepato-intestinal form of the disease, and 8 (61.5%) harboured light parasitic loads. In 1990, there were 27 (2.7%) infected individuals; less than half harboured light parasitic loads, with the predominance of moderate and heavy forms. Although our results indicate an improvement in the epidemiological situation of schistosomiasis in Sabugo, transmission of the disease in the locality is still active, especially among young males, and tends to be acquired during leisure activities.

Flow rate, pH and calcium concentration of saliva of children and adolescents with type 1 diabetes mellitus

Alterations in salivary parameters may increase the caries risk in diabetic children, but, contradictory data on this issue have been reported. The aims of this study were to compare salivary parameters (flow rate, pH and calcium concentration) between healthy and type 1 diabetes mellitus (T1DM) individuals. The sample consisted of 7- to 18-year-old individuals divided into two groups: 30 subjects with T1DM (group A) and 30 healthy control subjects (group B). Fasting glucose levels were determined. Unstimulated and stimulated saliva was collected. The pH of unstimulated saliva was measured with paper strips and an electrode. Calcium concentrations in stimulated saliva were determined with a selective electrode. Group A individuals had inadequate blood glucose control (HbA1C >9%), with means ± SD unstimulated salivary flow rate of 0.15 ± 0.1 mL/min compared to 0.36 ± 0.2 mL/min for group B (P < 0.01). Stimulated salivary flow rate was similar by both groups and above 2.0 mL/min. Saliva pH was 6.0 ± 0.8 for group A and significantly different from 7.0 ± 0.6 for group B (P < 0.01). Salivary calcium was 14.7 ± 8.1 mg/L for group A and significantly higher than 9.9 ± 6.4 mg/L for group B (P < 0.01). Except for elevated calcium concentrations in saliva, salivary parameters favoring caries such as low saliva pH and unstimulated salivary flow rate were observed in T1DM individuals.

Flow rate, pH and calcium concentration of saliva of children and adolescents with type 1 diabetes mellitus.

Alterations in salivary parameters may increase the caries risk in diabetic children, but, contradictory data on this issue have been reported. The aims of this study were to compare salivary parameters (flow rate, pH and calcium concentration) between healthy and type 1 diabetes mellitus (T1DM) individuals. The sample consisted of 7- to 18-year-old individuals divided into two groups: 30 subjects with T1DM (group A) and 30 healthy control subjects (group B). Fasting glucose levels were determined. Unstimulated and stimulated saliva was collected. The pH of unstimulated saliva was measured with paper strips and an electrode. Calcium concentrations in stimulated saliva were determined with a selective electrode. Group A individuals had inadequate blood glucose control (HbA(1C) >9%), with means +/- SD unstimulated salivary flow rate of 0.15 +/- 0.1 mL/min compared to 0.36 +/- 0.2 mL/min for group B (P < 0.01). Stimulated salivary flow rate was similar by both groups and above 2.0 mL/min. Saliva pH was 6.0 +/- 0.8 for group A and significantly different from 7.0 +/- 0.6 for group B (P < 0.01). Salivary calcium was 14.7 +/- 8.1 mg/L for group A and significantly higher than 9.9 +/- 6.4 mg/L for group B (P < 0.01). Except for elevated calcium concentrations in saliva, salivary parameters favoring caries such as low saliva pH and unstimulated salivary flow rate were observed in T1DM individuals.

[Abundance and infection of Biomphalaria glabrata with Schistosoma glabrata in the state of Rio de Janeiro, Brazil]. / Abundância e infecção do molusco Biomphalaria glabrata pelo Schistosoma mansoni no Estado do Rio de Janeiro, Brasil.

OBJECTIVES: To investigate the spatial distribution, abundance and natural schistosomiasis infection levels in the snail Biomphalaria glabrata, the intermediate host of Schistosoma mansoni in an area of the State of Rio de Janeiro, Brazil. METHODS: In the Pamparrão area, Sumidouro county, RJ, Brazil, snail captures were carried out every other month from June 1991 to November 1995. There were 23 collecting sites along the Pamparrão stream and its three tributaries (A, B and C). Captured snails were examined in the laboratory for infection. Data analyses were performed using Spearman coefficient (0.5% significance level) and Qui-square test. RESULTS: The abundance of B. glabrata was variable in both time and space. Most of the collection sites showed a negative correlation with rainfall. The tributary B had the highest and most steady infection rates (more than 25% in some sites). There were found significantly more infected snail in the dry season (chi2 = 20.08; p=0.001). CONCLUSIONS: B. glabrata population in the Pamparrão valley was negatively influenced by rainfall, especially at the Pamparrão stream. The dry season seems to promote the infection probably due to the lower water volume, increasing the chance of interaction between the intermediate host and the parasite.

Effects of cadmium on Euglena gracilis membrane lipids.

The toxic effects of cadmium (2 micrograms/ml) on membrane lipids and growth of Euglena gracilis were studied using autotrophic (AUTO), heterotrophic (DARK) and mixotrophic (LIGHT) cells. Cadmium caused inhibition of cellular proliferation (IC50 1.2 micrograms/ml) and morphological alterations which were most pronounced in chloroplasts. The chlorophyll content of LIGHT cadmium-treated cells was reduced 42.5%. Cadmium also caused an increase in protein and total lipid content per cell in all three cell types. Among the membrane lipids, cholesterol content was lower in cadmium-treated cells cultivated under illumination (AUTO: 0.40 +/- 0.02 vs 0.64 +/- 0.08 and LIGHT: 0.40 +/- 0.09 vs 0.53 +/- 0.01 microgram/10(5) cells). There were no changes in total phospholipid content, although cardiolipin content was altered in all three cell types, and in mixotrophic cells there was an increase in phosphatidylglycerol, a phospholipid typically found in chloroplasts. These results suggest that cadmium has an overall toxic effect on Euglena gracilis and that part of the effect can be ascribed to defects in the structure of chloroplasts and mitochondrial membranes.

Effects of cadmium on Euglena gracilis membrane lipids

The toxic effects of cadmium (2 mug/ml) on membrane lipids and growth of Euglena gracilis were studied using autotrophic (AUTO), heterotrophic (DARK) and mixotrophic (LIGHT) cells. Cadmium caused inhibition of cellular proliferation (IC50 1.2 mug/ml) and morphological alterations which were most pronounced in chloroplasts. The chlorophyll content of LIGHT cadmium-treated cells was reduced 42.5 per cent. Cadmium also caused an increase in protein and total lipid content per cell in all three cell types. Among the membrane lipids, cholesterol content was lower in cadmium-treated cells cultivated under illumination (AUTO: 0.40 ñ 0.02 vs 0.64 ñ 0.08 and LIGHT: 0.40 ñ 0.09 vs 0.53 ñ 0.01 mug/l0(5) cells). There were no changes in total phospholipid content, although cardiolipin content was altered in all three cell types, and in mixotrophic cells there was an increase in phosphatidylglycerol, a phospholipid typically found in chloroplasts. These results suggest that cadmium has an overall toxic effect on Euglena gracilis and that part of the effect can be ascribed to defects in the structure of chloroplasts and mitochondrial membranes.

Schistosomiasis in a low prevalence area: incomplete urbanization increasing risk of infection in Paracambi, RJ, Brazil.

The risk of schistosomiasis infection and heavy infection in the locality of Sabugo was evaluated in relation to housing in areas with different urbanization development and to residential supply with snail-infested water. Critical sanitary conditions were found in areas of incomplete urbanization, where healthy water supply sources were scarce, and draining of sewage, without previous treatment, was made directly to the water-bodies used for domestic and leisure activities, despite being Biomphalaria tenagophila snail breeding-places. Stool examinations (Kato-Katz and Lutz methods) showed prevalence of 2.9% mean intensity of 79 eggs per gram of stool and 47% of positive cases presenting intense infection. The use of snail-contaminated water for domestic purposes was considered a risk factor for infection. It is concluded that incomplete urbanization would facilitate transmission, probably enhancing the intensity of infection and that a low prevalence could hide a highly focal transmission. The relevance of these facts upon the efficiency of epidemiologic study methods and disease control planning are then discussed.

Rheotaxis of Biomphalaria glabrata on vertical substrates and its role in the recolonization of habitats treated with molluscicides

Os autores observaram exemplares de Biomphalaria glabrata subindo contra corrente em uma parede vertical de uma vala, constatando que os caramujos que apresentavam esse comportamento durante a aplicaçäo de miluscicida neste criadouro sobreviveram e, provavelmente, tiveram papel de repovoamento, que foi observado três meses após. Essas observaçöes suscitaram investigaçöes de campo e de laboratório, através das quais concluiram que: a) esta espécie é capaz de subir em superfícies verticais em situaçöes de campo e de laboratório; b) a corrente da água enquanto estímulo físico, é suficiente para desencadear esse comportamento (reotaxia); c) a reotaxia em superfícies verticais depende da existência de uma corrente de água necessariamente moderada; d) há indícios de que B. glabrata sofra habituaçäo em relaçäo à reotaxia em paredes verticais; e) a reotoxia em superfiícies verticais pode facilitar a dispersäo das populaçöes, devendo sua ocorrência ser considerada no planejamento das campanhas de controle da transmissäo da esquistossomose. Os autores apresentam propostas para evitar a manifestaçäo desse comportamento em algumas situaçöes de campo; f) a relaçäo entre a reotaxia e habituaçäo deve ser considerada um fator de agrupamento dos caramujos nas coleçöes d'água, que pode contribuir para sua localizaçäo

Studies on survival, biological activities and behavior of Biomphalaria glabrata, the host snail of Schistosomiasis, submitted to increased hydrostatic pressure: a technique

Para estudar mudanças na sobrevivência, atividades biológicas e comportamento de planorbídeos submetidos a aumento de pressäo hidrostática, desenvolvemos uma técnica que utiliza duas câmaras transparentes e um pistäo hidráulico. O aparelho permitiu a renovaçäo do meio líquido sem variaçöes substanciais na pressäo, eliminando assim os produtos de excreçäo e mantendo o nível de O2 disseolvido desejado, e desse modo permitido-nos avaliar o efeito da pressäo independente da ocorrência de anoxia. A pressäo foi mantida sem nenhum contato do meio líquido com o ar comprimido, situaçäo que produziu com relativa fidelildade o que ocorre na natureza, e assegurou a presença da mesma quantidade de gases nas duas câmaras de observaçäo. Biomphalaria glabrata foi capaz de sobreviver pelo menos 48 horas quando submetida a 49,02 x 10 Pa (equivalente a 48 m de profundidade) continuando a pôr massas ovíferas, e mostrando poucas modificaçöes comportamentais quando comparada com o grupo de controle

Studies on survival, biological activities and behavior of Biomphalaria glabrata, the host snail of schistosomiasis, submitted to increased hydrostatic pressure: a technique.

To study changes in survival, in biological activities and behavior of planorbids submitted to increased hydrostatic pressure, we developed a technique using two transparent chambers and a hydraulic piston. The apparatus permitted renewal of the liquid medium without substantial variations in pressure, thus eliminating excretion products and maintaining the desired O2 level and thereby permitting us to evaluate the effects of pressure independently of the occurrence of anoxia. Pressure was maintained without any contact of the liquid medium with compressed air, a situation which reproduced with relative fidelity what occurs in nature and assured the presence of the same amounts of gases in the two observation chambers (Control and Experimental). Biomphalaria glabrata was found to be able to survive at least 48 hours when submitted to 49.02 x 10(4) Pa (equivalent to a water depth of 48.8 m), continuing to lay egg masses and showing few behavioral changes when compared with the control group.

Rheotaxis of Biomphalaria glabrata on vertical substrates and its role in the recolonization of habitats treated with molluscicides.

The authors observed specimens of Biomphalaria glabrata climbing up the vertical wall of a ditch against the current. The snails that showed this behavior during application of a molluscicide in the breeding site survived and probably played a role in repopulation, which was observed three months later. These observations motivated field and laboratory investigations which led the authors to conclude that: a) this species is able to climb vertical surfaces both in field and laboratory situations; b) the current of water, as a physical stimulus, is sufficient to trigger this behavior (rheotaxis); c) rheotaxis on vertical surfaces depends on the presence of a necessarily moderate current; d) there are indications that B. glabrata may undergo habituation with respect to rheotaxis on vertical walls; e) the relationship between rheotaxis and habituation should be considered as a factor causing snail grouping in water bodies which may contribute to their localization in the field; f) rheotaxis on vertical surfaces may facilitate population dispersal, and its occurrence should be considered when campaigns for the control of schistosomiasis transmission are planned. The authors present some proposals to avoid the manifestation of this behavior in some field situations.

Behavior of Biomphalaria glabrata, the intermediate host snail of Schistosoma mansoni, at different depths in water in laboratory conditions

Investigamos a possibilidade de Biomphalaria glabrata deslocar-se para regiöes profundas, em três colunas de diferentes profundidades (1,10 m, 8,40 m e 10,40 m). Na coluna de 1,10 m evidenciamos que os deslocamentos podem ser de duas maneiras: 1) Quando o pé está em contato com o substrato: (i) deslizar para baixo; (ii) deslizar para cima; (iii) arrastar para baixo; (iiii) arrastar para cima. 2) Quando o pé näo está em contato com o substrato: (i) descida súbida sem saída de bolhas de ar; (ii) descida súbida com saída de bolhas de ar; (iii) subida súbita. Na coluna de 8,40 m em que havia alimento no fundo (grupo experimental) observamos que os caramujos permaneceram maior tempo nesta profundidade em relaçäo ao grupo que näo recebeu alimento (grupo de controle). Verificamos que o comportamento de deslizar é característico dos deslocamentos em torno de 0 a 1 m, tanto para subir como para descer. O comportamento de arrastar foi típico de subida dos caramujos que atingiram níveis mais profundos. Quando os caramujos arrastavam-se a concha ficava pendente como se estivesse mais pesada, o que pode estar relacionado à entrada de água na câmara pulmonar. Na coluna de 10,40 m verificamos que os caramujos deslizavam para baixo até 4 m ou apresentavam descidas súbitas até o fundo. As subidas se davam pelo padräo comportamental denominado arrastar, do fundo até a superfície. Nas colunas de 8,40 m e 10,40 m verificamos cópula, alimentaçäo e desova nos níveis mais profundos

Behavior of Biomphalaria glabrata, the intermediate host snail of Schistosoma mansoni, at different depths in water in laboratory conditions.

Using three columns of different depths (1.10m, 8.40m and 10.40m), we investigated the possibility of Biomphalaria glabrata moving towards deep regions. In the 1.10m column, we noted that locomotion can occur in two manners: 1) when the foot is in contact with the substrate: a) sliding descent; b) sliding ascent; c) creeping descent; d) creeping ascent, 2) when the foot is not in contact with the substrate: a) sudden descent without emission of air bules; b) sudden descent with emission of air bules; c) sudden ascent. In the 8.40m column containing food on the bottom (experimental group), the snails remained longer at this depth when compared to those of the group which received no food (control). The sliding behavior was characteristic of locomotion occurring at 0 to 1m both in upward and downward directions. Creeping behavior was typical for the ascent of the snails that reached deeper levels. When the snails were creeping, the shell remained hanging as if it were heavier, a fact that may have been due to water entering the pulmonary chamber. In the 10.40m column, the snails slid downward to a depth of 4m or descended suddenly all the way to the bottom. Ascent occurred by creeping from the bottom to the surface. In the 8.40m and 10.40m columns, copulation, feeding and oviposition occurred at the deepest levels.