Endogenous cholinergic tone modulates spontaneous network level neuronal activity in primary cortical cultures grown on multi-electrode arrays

Background Cortical cultures grown long-term on multi-electrode arrays (MEAs) are frequently and extensively used as models of cortical networks in studies of neuronal firing activity, neuropharmacology, toxicology and mechanisms underlying synaptic plasticity. However, in contrast to the predominantly asynchronous neuronal firing activity exhibited by intact cortex, electrophysiological activity of mature cortical cultures is dominated by spontaneous epileptiform-like global burst events which hinders their effective use in network-level studies, particularly for neurally-controlled animat (‘artificial animal’) applications. Thus, the identification of culture features that can be exploited to produce neuronal activity more representative of that seen in vivo could increase the utility and relevance of studies that employ these preparations. Acetylcholine has a recognised neuromodulatory role affecting excitability, rhythmicity, plasticity and information flow in vivo although its endogenous production by cortical cultures and subsequent functional influence upon neuronal excitability remains unknown. Results Consequently, using MEA electrophysiological recording supported by immunohistochemical and RT-qPCR methods, we demonstrate for the first time, the presence of intrinsic cholinergic neurons and significant, endogenous cholinergic tone in cortical cultures with a characterisation of the muscarinic and nicotinic components that underlie modulation of spontaneous neuronal activity. We found that tonic muscarinic ACh receptor (mAChR) activation affects global excitability and burst event regularity in a culture age-dependent manner whilst, in contrast, tonic nicotinic ACh receptor (nAChR) activation can modulate burst duration and the proportion of spikes occurring within bursts in a spatio-temporal fashion. Conclusions We suggest that the presence of significant endogenous cholinergic tone in cortical cultures and the comparability of its modulatory effects to those seen in intact brain tissues support emerging, exploitable commonalities between in vivo and in vitro preparations. We conclude that experimental manipulation of endogenous cholinergic tone could offer a novel opportunity to improve the use of cortical cultures for studies of network-level mechanisms in a manner that remains largely consistent with its functional role.

Inferring animal husbandry strategies in coastal zones through stable isotope analysis: new evidence from the Flemish coastal plain (Belgium, 1st-15th century AD)

In a proof-of-concept study, Britton et al. (2008) demonstrated that the isotopic composition of halophytic plants can be traced in the skeletal tissues of their animal consumers. Here we apply the method to domestic herbivore remains (n = 303) from nine archaeological sites in or near the Flemish coastal plain (Belgium), where, prior to embankments, salt-marshes offered extensive pasture grounds for domestic herbivores. The sites span a period of ∼1500 years (Roman to late medieval period), during which the coastal landscape was progressively transformed from little managed wetlands to a fully embanked polder area. The bulk collagen data show variations between sites and over time, which are consistent with this historical framework and are interpreted as reflecting environmental change and differences in animal management in the coastal plain throughout the late Holocene. The study demonstrates the immense value of faunal stable isotope analysis for characterising coastal husbandry strategies beyond the means of traditional zooarchaeological techniques.

Mediated action and sexual risk: Searching for 'culture' in discourses of homosexuality and AIDS prevention in China

This paper examines the concept of ‘culture’ and its relationship to HIV prevention. Culture is here seen as the interaction between human beings and the various ‘cultural tools’ they appropriate when taking action. Among these tools are ways of speaking which encode certain meanings, ideologies and social practices. When individuals take action with regard to AIDS, what they do is mediated through voices which they borrow strategically from their environ- ment. The textual tools that are available and the ways individuals adapt and combine them work to either limit or amplify their participation in HIV prevention. What are traditionally seen as ‘cultures’ or ‘sub-cultures’, or worse, ‘risk groups’, are, in this perspective, viewed as ‘communities of practice’, groups of individuals who share particular cultural tools and ways of using them. This conceptual framework is applied to recent discourses of homosexuality and AIDS prevention in China. An instance of ‘of� cial’ discourse in the form of an AIDS education pamphlet for ‘gays’ is analysed for the voices it contains and how these voices are strategically marshalled by the authors and mixed with other voices in ways which amplify participation in AIDS prevention for some and limit it for others. This ‘offical’ discourse is then compared to the discourse of homosexually active Chinese men recently interviewed in Beijing and Fuzhou to examine which of these of� cial voices and other voices they appropriate, and how they adapt these voices in responding to HIV.

Lymphocytes transfer [(14)C]-labeled fatty acids to skeletal muscle in culture; modulation by exercise

Previous studies have shown that lipids are transferred from lymphocytes (Ly) to different cell types including macrophages. enterocytes, and pancreatic beta cells in co-culture This study investigated whether [(14)C]-labeled fatty acids (FA) can be transferred from Ly to skeletal muscle (SM), and the effects of exercise on such phenomenon Ly obtained from exercised (EX) and control (C) male Wistar rats were preloaded with the [(14)C]-labeled free FA palmitic (PA), oleic (OA), linoleic (LA), or arachidonic (AA) Radioactively loaded Ly were then co-cultured with SM from the same Ly donor animals Substantial amounts of FA were transferred to SM being the profile PA = OA > AA > LA to the C group. and PA > OA > LA > AA to the EX group These FA were incorporated predominantly as phospholipids (PA = 66 75%: OA = 63 09%, LA = 43 86%, AA - 47 40%) in the C group and (PA = 63 99% OA = 52 72%, LA = 55 99%, AA = 63 40%) in the EX group Also in this group, the remaining radioactivity from AA, LA, and OA acids was mainly incorpoiated in structural and energetic lipids These results support the hypothesis that Ly are able to export lipids to SM in co-culture Furthermore. exercise modulates the lipid transference profile, and its incorporation on SM The overall significance of this phenomenon in vivo remains to be elucidated. Copyright (C) 2010 John Wiley & Sons, Ltd

Culture and exopolysaccharide production from sugarcane molasses by Gordonia polyisoprenivorans CCT 7137, isolated from contaminated groundwater in Brazil

Gordonia polyisoprenivorans CCT 7137 was isolated from groundwater contaminated with leachate in an old controlled landfill (Sauo Paulo, Brazil), and cultured in GYM medium at different concentrations of sugarcane molasses (2%, 6%, and 10%). The strain growth was analyzed by monitoring the viable cell counts (c.f.u. mL(-1)) and optical density and EPS production was evaluated at the end of the exponential phase and 24 h after it. The analysis of the viable cell counts showed that the medium that most favored bacterial growth was not the one that favored EPS production. The control medium (GYM) was the one that most favored the strain growth, at the maximum specific growth rate of 0.232 h(-1). Differences in bacterial growth when cultured at three different concentrations of molasses were not observed. Production of EPS, in all culture media used, began during the exponential phase and continued during the growth stationary phase. The highest total EPS production, after 24 h of stationary phase, was observed in 6% molasses medium (172.86 g L(-1)) and 10% (139.47 g L(-1)) and the specific total EPS production was higher in 10% molasses medium (39.03 x10(-11)g c.f.u.(-1)). After the exponential phase, in 2%, 6%, and 10% molasses media, a higher percentage of free exopolysaccharides (EPS) was observed, representing 88.4%, 62.4%, and 64.2% of the total, respectively. A different result was observed in pattern medium, which presented EPS made up of higher percentage of capsular EPS (66.4% of the total). This work is the first study on EPS production by G. polyisoprenivorans strain in GYM medium and in medium utilizing sugarcane molasses as the sole nutrient source and suggests its potential use for EPS production by G. polyisoprenivorans CCT 7137 aiming at application in biotechnological processes.

Density of sand flies (diptera: psychodidae) in domestic and wild animal shelters in an area of visceral leishmaniasis in the state of Rio Grande do Norte, Brazil

XIMENES, Maria de Fátima Freire de Melo; SOUZA, Maria de Fátima de; CASTELLON, Eloy Guilhermo. Density of sand flies (Diptera: Psychodidae) in domestic and wild animal shelters in an area of visceral Leishmaniasis in the State of Rio Grande do Norte, Brazil. Memorias do Instituto Oswaldo Cruz, Rio de Janeiro, v.94, n.4, p.427-432, jul./ago. 1999. Disponivel em: Acesso em: 4 out. 2010.

Solubilization of CaHPO4 and AlPO4 by Aspergillus niger in culture media with different carbon and nitrogen sources

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Growth and enzymatic responses of phytopathogenic fungi to glucose in culture media and soil

The effect of inoculation of Aspergillus flavus, Fusarium verticillioides, and Penicillium sp. in Dystrophic Red Latosol (DRL) and Eutroferric Red Latosol (ERL) soils with or without glucose on the total carbohydrate content and the dehydrogenase and amylase activities was studied. The fungal growth and spore production in culture medium with and without glucose were also evaluated. A completely randomized design with factorial arrangement was used. The addition of glucose in the culture medium increased the growth rate of A. flavus and Penicillium sp. but not of F. verticillioides. The number of spores increased 1.2 for F. verticillioides and 8.2 times for A. flavus in the medium with glucose, but was reduced 3.5 times for Penicillium sp. The total carbohydrates contents reduced significantly according to first and second degree equations. The consumption of total carbohydrates by A. flavus and Penicillium sp. was higher than the control or soil inoculated with F. verticillioides. The addition of glucose to soils benefited the use of carbohydrates, probably due to the stimulation of fungal growth. Dehydrogenase activity increased between 1.5 to 1.8 times (p <0.05) in soils with glucose and inoculated with the fungi (except F. verticillioides), in relation to soil without glucose. Amylase activity increased 1.3 to 1.5 times due to the addition of glucose in the soil. Increased amylase activity was observed in the DRL soil with glucose and inoculated with A. flavus and Penicillium sp. when compared to control.

Mineral Trioxide Aggregate-based Endodontic Sealer Stimulates Hydroxyapatite Nucleation in Human Osteoblast-like Cell Culture

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Repetitive DNA probe linked to sex chromosomes in hybrids between Neotropical fish Leporinus macrocephalus and Leporinus elongatus (Characiformes, Anostomidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Costs estimates and rentability of the guava culture (Psidium guajava L Paluma cv) in Monte Alto County, São Paulo State

This work intended to calculate the rentability of the guava culture Paluma cv. in Monte Alto region-SP, estimating the costs of the implantation, maintenance, production and the gross income of this culture in the first three years of cultivation. The production system utilized refers to the cultural treatments usually used in this culture in the region. The costs estimates were based on the total operational costs methodology used by the Agricultural Economics Institute (I.E.A.). The results obtained showed that the implantation and maintenance costs of the culture, in the two first years was R$7.402,3 l/alqueire*. Considering the possibility that in guava orchard, on irrigated conditions, reach in the fourthy year a productivity of 80t/alqueire and that the current price is R$0,20/kg, it is possible to estimate the gross income around of R$16.000,00/alqueire, resulting in a net income of R$9.497,06/alqueire.

MINERAL COMPOSITION of RAW MATERIAL, SUBSTRATE and FRUITING BODIES of Pleurotus ostreatus IN CULTURE

In a culture of a Pleurotus ostreatus (oyster mushroom) strain, macro and micronutrients of the raw material and the initial and spent substrates were evaluated. Substrates were formulated with sawdust from Simarouba amara Aubl. and Ochroma piramidale Cav. ex. Lam., crushed Bactris gasipaes Kunth and crushed Saccharum officinarum (sugar cane). Samples were solubilized by means of acid digestion (nitric-peridrol). Ca, Mg, Fe, Cu, Zn and Mn were determined by atomic absorption spectrophotometry, Na and K by atomic emission, and P by colorimetry. The mineral composition of the fruiting body varied with the substrates, which made possible the production of a fruiting body rich in K, P Mg and Fe. Potassium was the mineral with the highest content in the fruiting body in all substrates tested (36.83-42.18g.kg(-1)). There was an increase of protein and mineral content in the spent substrate in relation to the initial one.

Glucose-induced insulin secretion is impaired and insulin-induced phosphorylation of the insulin receptor and insulin receptor substrate-1 are increased in protein-deficient rats

Malnutrition is related to diabetes in tropical countries. In experimental animals, protein deficiency may affect insulin secretion. However, the effect of malnutrition on insulin receptor phosphorylation and further intracellular signaling events is not known. Therefore, we decided to evaluate the rate of insulin secretion and the early molecular steps of insulin action in insulin-sensitive tissues of an animal model of protein deficiency. Pancreatic islets isolated from rats fed a standard (17%) or a low (6%) protein diet were studied for their secretory response to increasing concentrations of glucose in the culture medium. Basal as well as maximal rates of insulin secretion were significantly lower in the islets isolated from rats fed a low protein diet. Moreover, the dose-response curve to glucose was significantly shifted to the right in the islets from malnourished rats compared with islets from control rats. During an oral glucose tolerance test, there were significantly lower circulating concentrations of insulin in the serum of rats fed a low protein diet in spite of no difference in serum glucose concentration between the groups, suggesting an increased peripheral insulin sensitivity. Immunoblotting and immunoprecipitation were used to study the phosphorylation of the insulin receptor and the insulin receptor substrate-1 as well as the insulin receptor substrate-1-p85 subunit of phosphatidylinositol 3-kinase association in response to insulin. Values were greater in hind-limb muscle from rats fed a low protein diet compared with controls. No differences were detected in the total amount of protein corresponding to the insulin receptor or insulin receptor substrate-1 between muscle from rats fed the two diets. Therefore, we conclude that a decreased glucose-induced insulin secretion in pancreatic islets from protein-malnourished rats is responsible, at least in part, for an increased phosphorylation of the insulin receptor, insulin receptor substrate-1 and its association with phosphatidylinositol 3-kinase. These might represent some of the factors influencing the equilibrium in glucose concentrations observed in animal models of malnutrition and undernourished subjects.

Costs estimates and rentability of the guava culture {psidium guajava l. patuma cv.) in monte alto county, Sao Paulo state

This work intended to calculate the rentability of the guava culture Paluma cv. in Monte Alto region-SP, estimating the costs of the implantation, maintenance, production and the gross income of this culture in the first three years of cultivation. The production system utilized refers to the cultural treatments usually used in this culture in the region. The costs estimates were based on the total operational costs methodology used by the Agricultural Economics Institute (I.E.A.). The results obtained showed that the implantation and maintenance costs of the culture, in the two first years was Rs7.402,31/alqueire. Considering the possibility that in guava orchard, on irrigated conditions, reach in the fourthy year a productivity of 80t/alqueire and that the current price is Rs0,20/kg, it is possible to estimate the gross income around of Rs16.000,00/alqueire, resulting in a net income of Rs9.497,06/alqueire.