Molecular determinants of vesicular storage and release of the gliotransmitter D-serine from cortical astrocytes

Neuron-astrocyte reciprocal communication at synapses has emerged as a novel signalling pathway in brain function. Astrocytes sense the level of synaptic activity and, in turn, influence its efficacy through the regulated release of 'gliotransmitters' such as glutamate, ATP or D-serine. A calcium-dependent exocytosis is proposed to drive the release of gliotransmitters but its existence is still debated. Over the last years, we have been studying the molecular determinants governing D-serine release from glia using different approaches. Using a novel bioassay for D-serine, we have been able to show that D-serine release occurs mainly through a calcium- and SNARE proteindependent mechanism just supporting the idea that this amino acid is released by exocytosis from glia. We next have pursued our exploration by confocal imaging and tracking of the exocytotic routes for Dserine- mediated gliotransmission and have shown that D-serine releasable pools are confined to synaptobrevin2/cellubrevin-bearing vesicles. To shed light onto the mechanisms controlling the storage and the release of gliotransmitters and namely D-serine, we have developed a new method for the immunoisolation of synaptobrevin 2- positive vesicles from rat cortical astrocytes in culture while preserving their content in gliotransmitters. The purified organelles are clear round shape vesicles of excellent purity with homogeneous size (40 nm) as judged by electron microscopy. Immunoblotting analysis revealed that isolated vesicles contain most of the major proteins already described for neuron-derived vesicles like synaptic vesicle protein 2 (SV2) and the proton pump H?-ATPase. In addition, we have analyzed the content for various amino acids of these vesicles by means of chiral capillary electrophoresis coupled to laser-induced fluorescence detection. The purified vesicles contain large amount of D-serine. We also detect peaks corresponding to unidentified compounds that may correspond to others amino acids. Postembedding immunogold labelling of the rat neocortex further revealed the expression of D-serine in astrocytes processes contacting excitatory synapses. Finally, we have examined the uptake properties for Dserine and glutamate inside the isolated glial vesicles. Our results provide significant support for the existence of an uptake system for D-serine in secretory glial vesicles and for the storage of chemical substances like D-serine and glutamate. 11th International Congress on Amino Acids, Peptides and Proteins 763 123

The fauna of phlebotomines (Diptera, Psychodidae) in different phytogeographic regions of the state of Maranhão, Brazil

Phlebotomine specimens were captured in domiciliary and forest environments in 47 municipalities between 1982 and 2005 with the aid of CDC light traps. A total of 91 species were found, of which four belonged to genus Brumptomyia and 87 to genus Lutzomyia, distributed among the following subgenera: Evandromyia (6), Lutzomyia (5), Micropygomyia (2), Nyssomyia (9), Pintomyia (2), Pressatia (3), Psathyromyia (6), Psychodopygus (14), Sciopemyia (4), Trichophoromyia (2), Viannamyia (2); species groups: Aragaoi (2), Baityi (1), Dreisbachi (1), Migonei (12), Oswaldoi (8), Pilosa (1), Saulensis (2), Verrucarum (4) and ungrouped (1). Species diversity was greatest in areas where there was dense evergreen seasonal forest (52 species), ombrophilous forest (31) and meridional cerrados (23) and lowest in areas with mixed forest (forest with babassu palms, cerrado and caatinga). The greatest similarity index was observed for restinga and open evergreen seasonal forest (J=0.48). Dense evergreen seasonal forest had greatest similarity with ombrophilous forest (J=0.38). The phlebotomine fauna was species rich and unevenly distributed in Maranhão, reflecting the phytogeographical complexity of the state, which is a result of the great variety of ecosystems and climate zones.

Phlebotomines (Diptera, Psychodidae) in the Speleological Province of the Ribeira Valley: 3. Serra district - area of hostels for tourists who visit the Parque Estadual do Alto Ribeira (PETAR), state of São Paulo, Brazil

Phlebotomines (Diptera, Psychodidae) in the Speleological Province of the Ribeira Valley: 3. Area of hostels for tourists who visit the Parque Estadual do Alto Ribeira (PETAR), state of São Paulo, Brazil. The study characterizes some ecological aspects of the phlebotomine fauna in an endemic area of cutaneous leishmaniasis (CL) situated in the Serra district, Iporanga municipality where the hostels for tourists visiting the PETAR are located. Captures were undertaken on a smallholding and a small farm situated near the hostels, monthly between January/2001 and December/2003 with automatic light traps (ALT) in pigsty, hen-house and veranda of a domicile at the two sites, and in peridomicile of the small farm also with black/white Shannon traps. With the ALT a total of 87,224 phlebotomines representing 19 species and also two hybrids of Nyssomyia intermedia (Lutz & Neiva) and Nyssomyia neivai (Pinto) and two anomalous specimens were captured. The standardized index species abundance was for Ny. intermedia = 1.0 and Ny. neivai = 0.935. The highest frequencies of the smallholding occurred in the pigsty, the Williams' mean/capture for Ny. intermedia being 63.7 specimens and for Ny. neivai 29.2, and on the small farm, in the hen-house, Ny. intermedia 402.6 and Ny. neivai 116.2. A total of 863 phlebotomines (Ny. intermedia: 75.4%; Ny. neivai: 24.3%) were captured with black/white Shannon traps; females of both species being predominant in the white trap. The high frequencies of Ny. intermedia and Ny. neivai, both implicated in CL transmission, indicate the areas presenting risk of the disease.

Diversity of Braconidae (Insecta, Hymenoptera) of the Parque Natural Municipal de Porto Velho, Rondonia, Brazil

Braconidae is a highly diversified family of Hymenoptera and usually known by their role in biological control both in agricultural and natural ecosystems. Despite of that, little is known about its diversity in the Amazon region. The present work inventoried the braconid fauna of an Open Ombrophylous Forest with Palm Trees of the Parque Natural Municipal de Porto Velho, RO. Insects were collect from June/2008 to May/2009 using six Malaise traps in different parts of the reserve. A total of 377 wasps were captured, 17 subfamilies and 56 genera identified. Braconinae, Microgastrinae, Doryctinae and Rogadinae subfamilies were very abundant, and also the genera Aleiodes, Bracon, Capitonius, Compsobracon, Heterospilus, Hymenochaonia, Opius, Pedinotus, Rogas and Stantonia. The calculated Shannon diversity index was 2.15 and 3.3 for subfamily and genera, respectively, which were, generally, higher than the values found for other regions in Brazil. Generally, parasitoids were more abundant during the rainy season. The present work contributes with new genera records and faunistic data of Braconidae in Rondonia State, western Amazon.

Orthoptera assemblages associated with macrophytes of floodplain lakes of the Paraná River

Orthoptera assemblages associated with macrophytes of floodplain lakes of the Paraná River. The Orthoptera assemblage composition varies considerably, depending on habitat type. This study examines the spatiotemporal relationship between plant diversity, hydrometric level, environmental variables and the Orthoptera richness and abundance in floodplain lakes connected permanently or temporarily with the main channel of the Paraná River. The grasshoppers were collected fortnightly (April 2006May 2007). A total of 17 species were recorded and classified according to their frequency of occurrence in constant (7), accessory (4), or accidental (6) species. In the two lakes, the greater species richness and abundance was recorded in summer, thereby coinciding with the highest water level of the Paraná River. The most significant correlation between the orthopteran richness and abundance was with the water level. The aquatic plant richness was significantly different between the lakes, but the vegetation was dominated by Eichhornia crassipes (Mart.) Solms. (Liliales, Pontederiaceae). The lake, which was connected permanently, presented the highest values of diversity and abundance, proving to be a more diverse assemblage. The beta diversity was higher in the temporary connected lake than in the permanently connected one. The orthopterans assemblages were different between the lakes, Cornops aquaticum and Tucayaca gracilis were the species that contributed more to the level of dissimilarity. C. aquaticum was more representative in the lake temporarily connected, while T. gracilis in the permanent connected one. The water level of the Paraná River and the connectivity of the floodplain lakes play an important role to explain the abundance and richness of their orthopteran assemblages.

Foraging strategies of the ant Ectatomma vizottoi (Hymenoptera, Formicidae)

Foraging strategies of the ant Ectatomma vizottoi (Hymenoptera, Formicidae). Foraging activity may be limited by temperature, humidity, radiation, wind, and other abiotic factors, all of which can affect energy costs during foraging. Ectatomma vizottoi's biology has only recently been studied, and no detailed information is available on its foraging patterns or diet in the field. For this reason, and because foraging activity is an important part of the ecological success of social insects, the present study aimed to investigate E. vizottoi's foraging strategies and dietary habits. First, we determined how abiotic factors constrained E. vizottoi's foraging patterns in the field by monitoring the foraging activity of 16 colonies on eight different days across two seasons. Second, we characterized E. vizottoi's diet by monitoring another set of 26 colonies during peak foraging activity. Our results show that E. vizottoi has foraging strategies that are similar to those of congeneric species. In spite of having a low efficiency index, colonies adopted strategies that allowed them to successfully obtain food resources while avoiding adverse conditions. These strategies included preying on other ant species, a foraging tactic that could arise if a wide variety of food items are not available in the environment or if E. vizottoi simply prefers, regardless of resource availability, to prey on other invertebrates and especially on other ant species.

Assessment of the in vitro synergy of daptomycin plus linezolid against multidrug-resistant enterococci

The widespread incidence of enterococci resistant to ampicillin, vancomycin and aminoglycosides, the first-line anti-enterococcal antibiotics, has made the treatment of severe enterococcal infections difficult and alternatives should be explored. We investigated the activity of daptomycin combined with linezolid against three Enterococcus faecalis and four Enterococcus faecium strains resistant to standard drugs used for therapy. Minimum inhibitory concentrations (MICs) were determined by the broth dilution method. Drug interactions were assessed by the checkerboard and time-kill methods. Synergy was defined by a fractional inhibitory concentration index (FICI) of ≤0.5 or a ≥2 log10 CFU/mL killing at 24 h with the combination in comparison with killing by the most active single agent. Indifference was defined by a FICI > 0.5-4.0 or a 1-2 log10 CFU/mL killing compared with the most active single agent. MICs of daptomycin were 2-4 μg/mL for E. faecalis and 2-8 μg/mL for E. faecium. MICs of linezolid were 1-2 μg/mL for all bacteria. In the checkerboard assay, five isolates showed synergism (FICI < 0.5) and two showed indifference (FICIs of 0.53 and 2). Killing studies revealed synergy of daptomycin plus linezolid against four isolates (2.2-3.7 log10 CFU/mL kill) and indifference (1.1-1.6 log10 CFU/mL kill) for the other three strains. Antagonism was not observed. In conclusion, the combination of daptomycin and linezolid had a synergistic or indifferent effect against multidrug-resistant enterococci. Additional studies are needed to explore the potential of this combination for severe enterococcal infections when first-line antibiotic combinations cannot be used.

The burden of musculoskeletal disorders in Catalonia

INTRODUCTION: Musculoskeletal disorders(MSDs)are extremely common and one of the major causes of disease burden around the world. The original Global Burden of Disease Study was commisioned by the World Bank in 1991 and established the DALY(Disability Ajusted Life Years) as an indicator of disease burden. The DALY calculation allows an assessment of not only mortality but also disability, in terms of time lived in health states worse than perfect health.OBJECTIVE: To estimate the burden of musculoskeletal disorders in Catalonia in 2010, through calculation of the DALY index.

Official Positions for FRAX® clinical regarding international differences from Joint Official Positions Development Conference of the International Society for Clinical Densitometry and International Osteoporosis Foundation on FRAX®.

Osteoporosis is a serious worldwide epidemic. Increased risk of fractures is the hallmark of the disease and is associated with increased morbidity, mortality and economic burden. FRAX® is a web-based tool developed by the Sheffield WHO Collaborating Center team, that integrates clinical risk factors, femoral neck BMD, country specific mortality and fracture data and calculates the 10 year fracture probability in order to help health care professionals identify patients who need treatment. However, only 31 countries have a FRAX® calculator at the time paper was accepted for publication. In the absence of a FRAX® model for a particular country, it has been suggested to use a surrogate country for which the epidemiology of osteoporosis most closely approximates the index country. More specific recommendations for clinicians in these countries are not available. In North America, concerns have also been raised regarding the assumptions used to construct the US ethnic specific FRAX® calculators with respect to the correction factors applied to derive fracture probabilities in Blacks, Asians and Hispanics in comparison to Whites. In addition, questions were raised about calculating fracture risk in other ethnic groups e.g., Native Americans and First Canadians. In order to provide additional guidance to clinicians, a FRAX® International Task Force was formed to address specific questions raised by physicians in countries without FRAX® calculators and seeking to integrate FRAX® into their clinical practice. The main questions that the task force tried to answer were the following: The Task Force members conducted appropriate literature reviews and developed preliminary statements that were discussed and graded by a panel of experts at the ISCD-IOF joint conference. The statements approved by the panel of experts are discussed in the current paper.

Growth and reproduction of the dolphinfish (Coryphaena equiselis and Coryphaena hippurus) in the Canary Islands, Central-East Atlantic (preliminary results)

The size, growth and spawning characteristics of pompano dolphin (N=150) and common dolphinfish (N=36) caught off the Canary Islands between May and September 1995 and between July and September 1996 were examined. Fork length (FL) of pompano dolphin was in the range 28.3-62.8 cm. In 1995, the mean length increased significantly from June to September. However, in 1996, the mean length was significantly larger in July than in September. The overall length-weight relationship was W=0.0287*FL2.774 (r=0.97), while these relationships by sex were as follows: W=0.031*FL2.758 (r=0.98) and W=0.0282*FL2.776 (r=0.97), for males and females respectively. Spawning takes place at the beginning of the Summer (June-July). All the individuals obtained showed developing gonads, but females showed a higher gonadosomatic index (GSI) than males. The highest GSI values were obtained in June (x- =3.10±1.73), and decreased progressively towards the end of the season (September), when the average of this index was x- = 1.86±0.87. Similarly, the condition index decreased significantly from June to September. The proportion of females was always significantly higher than males, except in July 1996 when it was 1:1. There was a high correspondence between growth rates determined by annuli scale interpretation and modal progression analysis. According to scale annuli interpretation, the individuals caught showed more than five age classes. However, there are doubts about age assignation from scales. Fork length of common dolphinfish was in the range of 76.5-103.0 cm. The length-weight relationships obtained for all the specimens caught was W=0.00095FL3.527 (r=0.96), while these relationships by sex were as follows: W=0.00398FL3.222 (r=0.94) and W=0.01656FL2.873 (r=0.91), for males and females respectively. Spawning probably takes place at the beginning of the Summer. All the individuals obtained showed developing gonads, although the GSI of females were higher than males. The highest GSI values were obtained in June (x- =5.50±2.17). In the same way, the condition index decreased from May to June. The proportion of females was always slightly higher than males (1:1.4), but the ratio was not significantly different from 1:1

Lipid (energy) reserves of european hake (Merluccius Merluccius) in the North-Western Mediterranean

This study analyses for the first time the lipid (energy) reserves of European hake (Merluccius merluccius) in the north-western Mediterranean from an ecophysiological perspective. Results show that there is a progressive accumulation of lipids in the liver of maturing hake -where the bulk of the fat is stored- as individuals grow. Results also indicate that female pre-spawners expend much energy on reproductive activities since they present lower liver lipid reserves than juveniles and maturing individuals. Furthermore, results show that female pre-spawners with higher lipid reserves in their livers had a higher amount of lipids in their ovaries, suggesting that maternal condition (spawner quality) may affect the reproductive potential of hake. Overall, the results of this study suggest that the analysis of liver lipid reserves during pre-spawning, along with the evaluation of the gonadosomatic index and the consideration of the reproductive stage, can contribute to improve the estimation of the reproductive potential of gadoid species such as hake

Validation of the Mini Nutritional Assessment short-form (MNA-SF): a practical tool for identification of nutritional status.

OBJECTIVE: To validate a revision of the Mini Nutritional Assessment short-form (MNA(R)-SF) against the full MNA, a standard tool for nutritional evaluation. METHODS: A literature search identified studies that used the MNA for nutritional screening in geriatric patients. The contacted authors submitted original datasets that were merged into a single database. Various combinations of the questions on the current MNA-SF were tested using this database through combination analysis and ROC based derivation of classification thresholds. RESULTS: Twenty-seven datasets (n=6257 participants) were initially processed from which twelve were used in the current analysis on a sample of 2032 study participants (mean age 82.3y) with complete information on all MNA items. The original MNA-SF was a combination of six questions from the full MNA. A revised MNA-SF included calf circumference (CC) substituted for BMI performed equally well. A revised three-category scoring classification for this revised MNA-SF, using BMI and/or CC, had good sensitivity compared to the full MNA. CONCLUSION: The newly revised MNA-SF is a valid nutritional screening tool applicable to geriatric health care professionals with the option of using CC when BMI cannot be calculated. This revised MNA-SF increases the applicability of this rapid screening tool in clinical practice through the inclusion of a "malnourished" category.

Between Equity and Differentiation: An Analytical Schema of the Social Function of Education

The educational sphere has an internal function relatively agreed by social scientists. Nonetheless, the contribution that educational systems provide to the society (i.e., their social function) does not have the same degree of consensus. Taking into consideration such theoretical precedent, the current article raises an analytical schema to grasp the social function of education considering a sociological perspective. Starting from the assumption that there is an intrinsic relationship between the internal and social functions of social systems, we suggest there are particular stratification determinants modifying the internal pedagogical function of education, which impact on its social function by creating simultaneous conditions of equity and differentiation. Throughout the paper this social function is considered a paradoxical mechanism. We highlight how this paradoxical dynamic is deployed in different structural levels of the educational sphere. Additionally, we discuss eventual consequences of this paradoxical social function for the inclusion possibilities that educational systems offer to individuals.