Influence of the dopaminergic system, CREB, and transcription factor-B on cocaine neurotoxicity

Cocaine is a widely used drug and its abuse is associated with physical, psychiatric and social problems. Abnormalities in newborns have been demonstrated to be due to the toxic effects of cocaine during fetal development. The mechanism by which cocaine causes neurological damage is complex and involves interactions of the drug with several neurotransmitter systems, such as the increase of extracellular levels of dopamine and free radicals, and modulation of transcription factors. The aim of this review was to evaluate the importance of the dopaminergic system and the participation of inflammatory signaling in cocaine neurotoxicity. Our study showed that cocaine activates the transcription factors NF-κB and CREB, which regulate genes involved in cellular death. GBR 12909 (an inhibitor of dopamine reuptake), lidocaine (a local anesthetic), and dopamine did not activate NF-κB in the same way as cocaine. However, the attenuation of NF-κB activity after the pretreatment of the cells with SCH 23390, a D1 receptor antagonist, suggests that the activation of NF-κB by cocaine is, at least partially, due to activation of D1 receptors. NF-κB seems to have a protective role in these cells because its inhibition increased cellular death caused by cocaine. The increase in BDNF (brain-derived neurotrophic factor) mRNA can also be related to the protective role of both CREB and NF-κB transcription factors. An understanding of the mechanisms by which cocaine induces cell death in the brain will contribute to the development of new therapies for drug abusers, which can help to slow down the progress of degenerative processes.

Effectiveness and biological compatibility of different generations of dentin adhesives

Besides possessing good mechanical properties, dental materials should present a good biological behavior and should not injure the involved tissues. Bond strength and biocompatibility are both highly significant properties of dentin adhesives. For that matter, these properties of four generations of adhesive systems (Multi-Purpose/Single Bond/SE Plus/Easy Bond) were evaluated.Eighty bovine teeth had their dentin exposed (500- and 200-mu m thickness). Adhesive was applied on the dentin layer of each specimen. Following that, the microshearing test was performed for all samples. A dentin barrier test was used for the cytotoxicity evaluation. Cell cultures (SV3NeoB) were collected from testing materials by means of 200- or 500-mu m-thick dentin slices and placed in a cell culture perfusion chamber. Cell viability was measured 24 h post-exposition by means of a photometrical test (MTT test).The best bonding performance was shown by the single-step adhesive Easy Bond (21 MPa, 200 mu m; 27 MPa, 500 mu m) followed by Single Bond (15.6 MPa, 200 mu m; 23.4 MPa, 500 mu m), SE Plus (18.2 MPa, 200 mu m; 20 MPa, 500 mu m), and Multi-Purpose (15.2 MPa, 200 mu m; 17.9 MPa, 500 mu m). Regarding the cytotoxicity, Multi-Purpose slightly reduced the cell viability to 92 % (200 mu m)/93 % (500 mu m). Single Bond was reasonably cytotoxic, reducing cell viability to 71 % (200 mu m)/64 % (500 mu m). The self-etching adhesive Scotchbond SE decreased cell viability to 85 % (200 mu m)/71 % (500 mu m). Conversely, Easy Bond did not reduce cell viability in this test, regardless of the dentin thickness.Results showed that the one-step system had the best bond strength performance and was the least toxic to pulp cells. In multiple-step systems, a correct bonding technique must be done, and a pulp capping strategy is necessary for achieving good performance in both properties.The study showed a promising system (one-step self-etching), referring to it as a good alternative for specific cases, mainly due to its technical simplicity and good biological responses.

Assessing melatonin and its oxidative metabolites amounts in biological fluid and culture medium by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS)

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

Biological control of phytophagous arthropods in the physic nut tree Jatropha curcas L. in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biological associations of color variation in the Indo-Pacific swimming crab Charybdis hellerii

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Immunolocalization of myosin Va in the developing nervous system of embryonic chicks

Myosins are molecular motors associated with the actin cytoskeleton that participate in the mechanisms of cellular motility. During the development of the nervous system, migration of nerve cells to specific sites, extension of growth cones, and axonal transport are dramatic manifestations of cellular motility. We demonstrate, via immunoblots, the expression of myosin Va during early stages of embryonic development in chicks, extending from the blastocyst period to the beginning of the fetal period. The expression of myosin Va in specific regions and cellular structures of the nervous system during these early stages was determined by immunocytochemistry using a polyclonal antibody. Whole mounts of chick embryos at 24-30-h stages showed intense immunoreactivity of the neural tube in formation along its full extent. Cross-sections at these stages of development showed strong labeling in neuroepithelial cells at the basal and apical regions of the neural tube wall. Embryos at more advanced periods of development (48h and 72 h) showed distinctive immunolabeling of neuroepithelial cells, neuroblasts and their cytoplasmic extensions in the mantle layer of the stratified neural tube wall, and neuroblasts and their cytoplasmic extensions in the internal wall of the optic cup, as well as a striking labeling of cells in the apparent nuclei of cranial nerves and budding fibers. These immunolocalization studies indicate temporal and site-specific expression of myosin Va during chick embryo development, suggesting that myosin Va expression is related to recruitment for specific cellular tasks.

Chemical and biological characterisation of Machaerium hirtum (Vell.) Stellfeld: absence of cytotoxicity and mutagenicity and possible chemopreventive potential

Machaerium hirtum (Vell.) Stellfeld (M.hirtum) is a plant known as 'jacarandá-bico-de-pato' whose bark is commonly used against diarrhea, cough and cancer. The aim of this study was to phytochemically characterise the hydroethanolic extract of this plant, investigate its antimutagenic activities using the Ames test and evaluate its effects on cell viability, genomic instability, gene expression and cell protection in human hepatocellular carcinoma cells (HepG2). Antimutagenic activity was assessed by simultaneous pre- and post-treatment with direct and indirect mutagens, such as 4-nitro-o-phenylenediamine (NPD), mitomycin C (MMC), benzo[a]pyrene (B[a]P) and aflatoxin B1 (AFB1), using the Ames test, cytokinesis blocking micronucleus and apoptosis assays. Only 3 of the 10 concentrations evaluated in the MTT assay were cytotoxic in HepG2 cells. Micronucleated or apoptotic cells were not observed with any of the tested concentrations, and there were no mutagenic effects in the bacterial system. However, the Nuclear Division Index and flow cytometry data showed a decrease in cell proliferation. The extract showed an inhibitory effect against direct (NPD) and indirect mutagens (B[a]P and AFB1). Furthermore, pre- and post-treated cells showed significant reduction in the number of apoptotic and micronucleated cells. This effect is not likely to be associated with the modulation of antioxidant genes, as shown by the RT-qPCR results. Six known flavonoids were identified in the hydroethanolic extract of Machaerium hirtum leaves, and their structures were elucidated by spectroscopic and spectrophotometric methods. The presence of the antioxidants apigenin and luteolin may explain these protective effects, because these components can inhibit the formation of reactive species and prevent apoptosis and DNA damage. In conclusion, the M.hirtum extract showed chemopreventive potential and was not hazardous at the tested concentrations in the experiments presented here. Moreover, this extract should be investigated further as a chemopreventive agent.

Surfactant-based transdermal system for fluconazole skin delivery

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

Limnology of a lateral lagoon system connected to a Neotropical reservoir (Rosana reservoir, São Paulo/Paraná, Brazil)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biological control of root rot caused by Pythium aphanidermatum and growth promotion in hydroponic cucumber with microorganisms from mangrove

The prospection of biological control agents in similar environments to the microbe application improves the chances of microorganisms establishment added to the environment. The low survival of these beneficial microorganisms added to hydroponic environment is a problem for the growth promotion and root rot biological control success in hydroponic crops. Because of the environmental similarity between hydroponic systems and mangrove ecosystems, the aim of this work was to evaluate the ability of mangrove microbes to control root rot caused by Pythium aphanidermatum and to improve plant growth in hydroponic cucumbers. Among the 28 strains evaluated for disease control in small-hydroponic system using cucumber seedlings, Gordonia rubripertincta SO-3B-2 alone or in combination with Pseudomonas stutzeri (MB-P3A- 49, MB-P3-C68 and SO-3L-3), and Bacillus cereus AVIC-3-6 increased the seedlings survival and were subsequently evaluated in hydroponic cucumbers in a greenhouse. Bacillus cereus AVIC-3-6 protected the plants from stunting caused by the pathogen and Gordonia rubripertincta SO-3B-2 and Pseudomonas stutzeri MB-P3A-49 increased the plant growth. We concluded that microorganisms from mangroves are useful as biocontrol agents and for improving plant growth in hydroponic crops.

Biological control of the bronze bug, Thaumastocoris peregrinus, in eucalyptus plantations in Brazil

The bronze bug Thaumastocoris peregrinus (Hemiptera: Thaumastocoridae) was detected in Brazil in 2008 and infested >180 000 ha of eucalyptus plantations in 2011. The bronze bug can cause a reduction of 10–15% in wood productivity after 2 years of heavy infestation. Although there is not an effective control method known, biological control is the main control strategy studied. An exotic egg parasitoid, Cleruchoides noackae (Hymenoptera: Mymaridae), was imported from Australia in 2012, reared in a laboratory and released in three Brazilian regions. Parasitoids were recovered at release points after 20–30 d. In 2013, preliminary evaluations demonstrated parasitoid establishment in these areas, and the parasitoid was recovered in adjacent areas after 1 year of release. Bioassays confi rmed egg parasitism of 15–20% by C. noackae. Other native natural enemies were studied. We found green lacewing Chrysoperla externa and predatory bugs Supputius cincticeps and Atopozelus opsimus preying on nymphs and/or adults of T. peregrinus. Another promising possibility is entomopathogenic fungi. Commercial formulations of Beauveria bassiana were tested with success in lab and fi eld conditions. Fusarium proliferatum and Paecilomyces cateniannulatus caused mortality of T. peregrinus in natural epizooties. After 5 years of research, it is possible to develop an integrated pest management system (IPM) for eucalyptus plantations based on biocontrol strategy for bronze bug.

Assessing the gain of biological data integration in gene networks inference

Background: A current challenge in gene annotation is to define the gene function in the context of the network of relationships instead of using single genes. The inference of gene networks (GNs) has emerged as an approach to better understand the biology of the system and to study how several components of this network interact with each other and keep their functions stable. However, in general there is no sufficient data to accurately recover the GNs from their expression levels leading to the curse of dimensionality, in which the number of variables is higher than samples. One way to mitigate this problem is to integrate biological data instead of using only the expression profiles in the inference process. Nowadays, the use of several biological information in inference methods had a significant increase in order to better recover the connections between genes and reduce the false positives. What makes this strategy so interesting is the possibility of confirming the known connections through the included biological data, and the possibility of discovering new relationships between genes when observed the expression data. Although several works in data integration have increased the performance of the network inference methods, the real contribution of adding each type of biological information in the obtained improvement is not clear. Methods: We propose a methodology to include biological information into an inference algorithm in order to assess its prediction gain by using biological information and expression profile together. We also evaluated and compared the gain of adding four types of biological information: (a) protein-protein interaction, (b) Rosetta stone fusion proteins, (c) KEGG and (d) KEGG+GO. Results and conclusions: This work presents a first comparison of the gain in the use of prior biological information in the inference of GNs by considering the eukaryote (P. falciparum) organism. Our results indicates that information based on direct interaction can produce a higher improvement in the gain than data about a less specific relationship as GO or KEGG. Also, as expected, the results show that the use of biological information is a very important approach for the improvement of the inference. We also compared the gain in the inference of the global network and only the hubs. The results indicates that the use of biological information can improve the identification of the most connected proteins.

Evaluation of Microbicidal Activity of Oxygen-Containing Plasmas using Biological Monitors with Different Lumen Calibers

New technologies and sterilization agents for heat-sensitive materials are under intense investigation. Plasma sterilization, an atoxic low-temperature substitute for conventional sterilization, uses various gases that are activated by an electrical discharge, generating reactive species that promote lethality in microorganisms. Here, assays were performed using pure O-2 and O-2 + H2O2 mixture gas plasmas against a standard load of Bacillus atrophaeus spores inoculated on glass carriers inside PVC catheters. The sterilization efficiency was studied as a function of plasma system (reactive ion etching or inductively coupled plasma), biological monitor lumen diameter, gas, radio frequency power, and sub-lethal exposition time. After sterilization, the biological monitors were disassembled and the surviving bacteria were grown in trypticase soy broth using the most probable number technique. Plasma antimicrobial activity depended on the catheter's internal diameter and radio frequency powers. The N-2 + H2O2 mixture exhibited higher microbial efficacy than pure N-2 in both plasma systems.

Gene expression profiles displayed by peripheral blood mononuclear cells from patients with type 2 diabetes mellitus focusing on biological processes implicated on the pathogenesis of the disease

Patients with type 2 diabetes mellitus (T2DM) exhibit insulin resistance associated with obesity and inflammatory response, besides an increased level of oxidative DNA damage as a consequence of the hyperglycemic condition and the generation of reactive oxygen species (ROS). In order to provide information on the mechanisms involved in the pathophysiology of T2DM, we analyzed the transcriptional expression patterns exhibited by peripheral blood mononuclear cells (PBMCs) from patients with T2DM compared to non-diabetic subjects, by investigating several biological processes: inflammatory and immune responses, responses to oxidative stress and hypoxia, fatty acid processing, and DNA repair. PBMCs were obtained from 20 T2DM patients and eight non-diabetic subjects. Total RNA was hybridized to Agilent whole human genome 4x44K one-color oligo-microarray. Microarray data were analyzed using the GeneSpring GX 11.0 software (Agilent). We used BRB-ArrayTools software (gene set analysis - GSA) to investigate significant gene sets and the Genomica tool to study a possible influence of clinical features on gene expression profiles. We showed that PBMCs from T2DM patients presented significant changes in gene expression, exhibiting 1320 differentially expressed genes compared to the control group. A great number of genes were involved in biological processes implicated in the pathogenesis of T2DM. Among the genes with high fold-change values, the up-regulated ones were associated with fatty acid metabolism and protection against lipid-induced oxidative stress, while the down-regulated ones were implicated in the suppression of pro-inflammatory cytokines production and DNA repair. Moreover, we identified two significant signaling pathways: adipocytokine, related to insulin resistance; and ceramide, related to oxidative stress and induction of apoptosis. In addition, expression profiles were not influenced by patient features, such as age, gender, obesity, pre/post-menopause age, neuropathy, glycemia, and HbA(1c) percentage. Hence, by studying expression profiles of PBMCs, we provided quantitative and qualitative differences and similarities between T2DM patients and non-diabetic individuals, contributing with new perspectives for a better understanding of the disease. (C) 2012 Elsevier B.V. All rights reserved.

The chemiluminescent peroxyoxalate system: state of the art almost 50 years from its discovery

Almost fifty years after the discovery of the peroxyoxalate reaction by E. A. Chandross in the early nineteen sixties, this review article intends to give a general overview on mechanistic aspects of this system and to describe the principles of its analytical application. After a short general introduction on the principles of chemiluminescence and the history of peroxyoxalate discovery, mechanistic aspects of high-energy intermediate formation, its structure and its reaction with an activator in the peroxyoxalate system are discussed. Finally, analytical applications of peroxyoxalate chemiluminescence are exemplified using representative recent examples, including oxalic acid detection in biological samples.