Cytotoxic isolates of Helicobacter pylori from Peptic Ulcer Diseases decrease K(+)-dependent ATPase Activity in HeLa cells

BACKGROUND: Helicobacter pylori is a Gram negative bacterium that plays a central role in the etiology of chronic gastritis and peptic ulcer diseases. However, not all H. pylori positive cases develop advanced disease. This discriminatory behavior has been attributed to the difference in virulence of the bacteria. Among all virulence factors, cytotoxin released by H. pylori is the most important factor. In this work, we studied variation in H. pylori isolates from Indian dyspeptic patients on the basis of cytotoxin production and associated changes in K(+)-dependent ATPase (one of its targets) enzyme activity in HeLa cells. METHODS: The patients were retrospectively grouped on the basis of endoscopic and histopathological observation as having gastritis or peptic ulcer. The HeLa cells were incubated with the broth culture filtrates (BCFs) of H. pylori isolates from patients of both groups and observed for the cytopathic effects: morphological changes and viability. In addition, the K(+)-dependent ATPase activity was measured in HeLa cells extracts. RESULTS: The cytotoxin production was observed in 3/7 (gastritis) and 4/4 (peptic ulcer) H. pylori isolates. The BCFs of cytotoxin producing H. pylori strains reduced the ATPase activity of HeLa cells to 40% of that measured with non-cytotoxin producing H. pylori strains (1.33 μmole Pi/mg protein and 3.36 μmole Pi/mg protein, respectively, p < 0.05). The decreased activity of ATPase enzyme or the release of cytotoxin also correlated with the increased pathogenicity indices of the patients. CONCLUSIONS: Our results suggest that the isolation of cytotoxic H. pylori is more common in severe form of acid peptic diseases (peptic ulcer) than in gastritis patients from India. Also the cytotoxin released by H. pylori impairs the ion-transporting ATPase and is a measure of cytotoxicity

Signal transduction-related responses to phytohormones and environmental challenges in sugarcane

BACKGROUND: Sugarcane is an increasingly economically and environmentally important C4 grass, used for the production of sugar and bioethanol, a low-carbon emission fuel. Sugarcane originated from crosses of Saccharum species and is noted for its unique capacity to accumulate high amounts of sucrose in its stems. Environmental stresses limit enormously sugarcane productivity worldwide. To investigate transcriptome changes in response to environmental inputs that alter yield we used cDNA microarrays to profile expression of 1,545 genes in plants submitted to drought, phosphate starvation, herbivory and N(2)-fixing endophytic bacteria. We also investigated the response to phytohormones (abscisic acid and methyl jasmonate). The arrayed elements correspond mostly to genes involved in signal transduction, hormone biosynthesis, transcription factors, novel genes and genes corresponding to unknown proteins. RESULTS: Adopting an outliers searching method 179 genes with strikingly different expression levels were identified as differentially expressed in at least one of the treatments analysed. Self Organizing Maps were used to cluster the expression profiles of 695 genes that showed a highly correlated expression pattern among replicates. The expression data for 22 genes was evaluated for 36 experimental data points by quantitative RT-PCR indicating a validation rate of 80.5% using three biological experimental replicates. The SUCAST Database was created that provides public access to the data described in this work, linked to tissue expression profiling and the SUCAST gene category and sequence analysis. The SUCAST database also includes a categorization of the sugarcane kinome based on a phylogenetic grouping that included 182 undefined kinases. CONCLUSION: An extensive study on the sugarcane transcriptome was performed. Sugarcane genes responsive to phytohormones and to challenges sugarcane commonly deals with in the field were identified. Additionally, the protein kinases were annotated based on a phylogenetic approach. The experimental design and statistical analysis applied proved robust to unravel genes associated with a diverse array of conditions attributing novel functions to previously unknown or undefined genes. The data consolidated in the SUCAST database resource can guide further studies and be useful for the development of improved sugarcane varieties

Bupivacaine in alginate and chitosan nanoparticles: an in vivo evaluation of efficacy, pharmacokinetics, and local toxicity

C&iacute;ntia Maria Saia Cereda,1 Daniel Sebbe Mecatti,2 Juliana Zampoli Boava Papini,1 Diego Val&eacute;rio Bueno,2 Michelle Franz-Montan,3 Thalita Rocha,2 Jos&eacute; Pedrazzoli J&uacute;nior,2 Eneida de Paula,4 Daniele Ribeiro de Ara&uacute;jo,5 Renato Grillo,6 Leonardo Fernandes Fraceto,7 Silvana Aparecida Calafatti,2 Giovana Radomille Tofoli1 1Institute and Research Center S&atilde;o Leopoldo Mandic, Campinas, S&atilde;o Paulo, Brazil; 2UNIFAG, S&atilde;o Francisco University, Bragan&ccedil;a Paulista, S&atilde;o Paulo, Brazil; 3Department of Physiological Sciences, University of Campinas, Piracicaba, S&atilde;o Paulo, Brazil; 4Department of Biochemistry and Tissue Biology, University of Campinas, Campinas, São Paulo, Brazil; 5Human and Natural Science Centre, Federal University of ABC, Santo Andr&eacute;, S&atilde;o Paulo, Brazil; 6Department of Physics and Chemistry, School of Engineering, S&atilde;o Paulo State University (UNESP), Ilha Solteira, S&atilde;o Paulo, Brazil; 7Department of Environmental Engineering, S&atilde;o Paulo State University (UNESP), Sorocaba, S&atilde;o Paulo, Brazil Objective: This study reports a preclinical evaluation of an alginate/chitosan nanoparticle formulation containing NovaBupi&reg;, a racemic bupivacaine (BVC) containing 25% dextrobupivacaine and 75% levobupivacaine. Methods: New Zealand White rabbits (n=6) received intraoral or intrathecal injections of BVC 0.5% or BVC 0.5%-loaded alginate&ndash;chitosan nanoparticles (BVCALG). BVC plasma levels and pharmacokinetic parameters were determined in blood samples of these rabbits. An infraorbital nerve blockade was performed in male Wistar rats (n=7) with the same formulations and the vehicle (NPALG). Histological evaluation of local toxicity after 6 hours and 24 hours of the treatments was performed in rats&rsquo; (n=6) oral tissues. Results: No statistically significant difference was observed between plasma concentrations and pharmacokinetic parameters (p&gt;0.05) after intraoral injections. However, after intrathecal injection BVCALG changed approximately three times the values of volume of distribution and area under the curve (AUC0&ndash;t; p&lt;0.05). The total analgesic effect of BVC after infraorbital nerve blockade was improved by 1.4-fold (p&lt;0.001) with BVCALG. BVC and BVCALG did not induce significant local inflammatory reaction. Conclusion: The encapsulation of BVC prolongs the local anesthetic effect after infraorbital nerve blockade and altered the pharmacokinetics after intrathecal injection. Keywords: local anesthetics, bupivacaine, polymeric nanoparticle, drug delivery, preclinical stud

Identification of Sensitive Enzymes in the Photosynthetic Carbon Metabolism

Understanding and optimizing the CO2 fixation process would allow human beings to address better current energy and biotechnology issues. We focused on modeling the C3 photosynthetic Carbon metabolism pathway with the aim of identifying the minimal set of enzymes whose biotechnological alteration could allow a functional re-engineering of the pathway. To achieve this result we merged in a single powerful pipe-line Sensitivity Analysis (SA), Single- (SO) and Multi-Objective Optimization (MO), and Robustness Analysis (RA). By using our recently developed multipurpose optimization algorithms (PAO and PMO2) here we extend our work exploring a large combinatorial solution space and most importantly, here we present an important reduction of the problem search space. From the initial number of 23 enzymes we have identified 11 enzymes whose targeting in the C3 photosynthetic Carbon metabolism would provide about 90% of the overall functional optimization. Both in terms of maximal CO2 Uptake and minimal Nitrogen consumption, these 11 sensitive enzymes are confirmed to play a key role. Finally we present a RA to confirm our findings. © 2012 Springer Science+Business Media, LLC

Gender Differences in Positive and Negative Feelings Between Adolescents and their Fathers and Mothers

Given the connections between feelings and well-being within the family, we addressed gender differences and similarities of feelings within the family. We explored adolescents’, mothers’, and fathers’ positive and negative affect within a community sample of 224 families from the southeast of the United States. Adolescents ranged in age from 12 to 19 years. Findings suggest more gender differences in positive and negative affect for parental gender (i.e., mothers versus fathers) than for adolescent gender (i.e., boys versus girls). There were more consistent associations within the family for positive affect as opposed to negative affect. We discussed the results in the context of gender differences within the family within the family systems theoretical perspective