Qualitative Analysis of the Carbohydrate Composition of Apolipoprotein H

The specific binding of digoxigenin-labeled lectins to carbohydrate moieties is used to characterize the carbohydrate chains bound to apolipoprotein H. Our results show that apolipoprotein H is rich in sialic acid linked (2–6) to galactose or N-acetylgalactosamine. Sialic acid is not (2–3)-linked to galactose. Galactose is (1–4)-linked to N-acetylglucosamine and (1–3)-linked to N-acetylgalactosamine. High-mannose N-glycan chains are barely detectable. After N-glycosidase F treatment the molecular weight is substantially reduced. The main band is 32,500 daltons. Carbohydrate O-linked chains, which are mainly represented by sialic acid, are (2–6)-linked to galactose or N-acetylgalactosamine. Galactose is also organized in O-linked chains and it is (1–4)-linked to N-acetylglucosamine and (1–3)-linked to acetylgalactosamine. Biochemical analysis of carbohydrate structures reveals that no specific carbohydrate complex is bound to a single isoform.     

Evaluation of the phenotypic characteristics of coliform bacteria recovered with two methods: the European Drinking Water Directive reference method and the Colilert 18/Quanti-Tray™ system

Summary The taxonomy of the species belonging to Enterobacteriaceae has undergone a series of revisions. As a consequence, the new classification has caused the analytical detection methods to be updated. According to the European Drinking Water Directive 98/83/CE coliforms/Escherichia coli have to be determined with the ISO 9308-1 reference method. Many technical drawbacks of the procedure as well as limitations regarding the recent taxonomy of coliforms have been pointed out by laboratories working in water quality control. In our investigation, water was analyzed in parallel with the reference method and the rapid Colilert 18/Quanti-Tray™ system. Phenotypic characteristics of isolates were recorded for the verification of the response of the two methods to the new taxonomic approach. Results obtained with the ISO standard confirmed the limitations of the test. In fact, in addition to difficulties linked to the readability of results, it failed to detect a significant proportion of coliforms and E. coli in water. Furthermore, it allowed the growth of microorganisms belonging to other groups. The Colilert 18/Quanti-Tray™ system detected a qualitatively and quantitatively higher number of target microorganisms. It also provided results in a shorter time, allowing the simultaneous detection of E. coli and coliforms with no further confirmation steps.     

The inhibitory activity of wine yeast starters on malolactic bacteria

Malolactic fermentation is a process that is influenced by various factors that can inhibit the growth of the malolactic bacteria. Inhibitory metabolites produced by yeast may have an important role in the correct development of malolactic fermentation. For these reasons, we have investigated the effects of such metabolites on the growth of malolactic bacteria under different environmental conditions, to aid in our understanding of the significance of these interactions in the wine-making environment. Our screening methods to detect interactions between yeast and malolactic bacteria showed a variable and wide diffusion of yeast inhibitory activity on the growth of the malolactic bacteria. However, this first approach to determine this inhibitory activity of yeast gave an overestimation when compared to the results obtained under actual wine-making conditions. The evaluation of malic acid consumption indicated that under inhibitory conditions a partial L-malic acid degradation was seen, indicating that the malolactic activity continued without bacterial growth. However, these yeast-inhibiting effects in addition to other environmental factors could cause a complete failure of malolactic fermentation.     

Mountain cheese factory wastewater treatment with the use of a hybrid constructed wetland

This paper describes the activity period of an experimental hybrid wetland system placed in a cold climate region. The aim is to determine the efficiency of the system in reducing TSS, BOD₅, COD and other pollutants. The constructed wetland consists of a fat-removal unit and a basin for the storage and the distribution of the wastewater which precedes three phytoremediation beds: the first two are parallel and they work as submerged vertical flow wetland with gravel medium for an area of 180 m²; the last is a submerged horizontal flow wetland with sand medium and an area of 360 m². The CW was designed to process a total estimated BOD₅ loading rate of about 24 g m⁻² d⁻¹, which was less than half of the average actual loading rate. The wastewater treatment did not meet the required Italian law outflow limits, most likely due to BOD₅ overloading.     

Clinical Evaluation of Rega 8: An Updated Genotypic Interpretation System That Significantly Predicts HIV-Therapy Response

Introduction

Clinically evaluating genotypic interpretation systems is essential to provide optimal guidance in designing potent individualized HIV-regimens. This study aimed at investigating the ability of the latest Rega algorithm to predict virological response on a short and longer period.

Materials & Methods

9231 treatment changes episodes were extracted from an integrated patient database. The virological response after 8, 24 and 48 weeks was dichotomized to success and failure. Success was defined as a viral load below 50 copies/ml or alternatively, a 2 log decrease from the baseline viral load at 8 weeks. The predictive ability of Rega version 8 was analysed in comparison with that of previous evaluated version Rega 5 and two other algorithms (ANRS v2011.05 and Stanford HIVdb v6.0.11). A logistic model based on the genotypic susceptibility score was used to predict virological response, and additionally, confounding factors were added to the model. Performance of the models was compared using the area under the ROC curve (AUC) and a Wilcoxon signed-rank test.

Results

Per unit increase of the GSS reported by Rega 8, the odds on having a successful therapy response on week 8 increased significantly by 81% (OR = 1.81, CI = [1.76–1.86]), on week 24 by 73% (OR = 1.73, CI = [1.69–1.78]) and on week 48 by 85% (OR = 1.85, CI = [1.80–1.91]). No significant differences in AUC were found between the performance of Rega 8 and Rega 5, ANRS v2011.05 and Stanford HIVdb v6.0.11, however Rega 8 had the highest sensitivity: 76.9%, 76.5% and 77.2% on 8, 24 and 48 weeks respectively. Inclusion of additional factors increased the performance significantly.

Conclusion

Rega 8 is a significant predictor for virological response with a better sensitivity than previously, and with rules for recently approved drugs. Additional variables should be taken into account to ensure an effective regimen.     

Modeling multi-sensory feedback control of zebrafish in a flow

Understanding how animals navigate complex environments is a fundamental challenge in biology and a source of inspiration for the design of autonomous systems in engineering. Animal orientation and navigation is a complex process that integrates multiple senses, whose function and contribution are yet to be fully clarified. Here, we propose a data-driven mathematical model of adult zebrafish engaging in counter-flow swimming, an innate behavior known as rheotaxis. Zebrafish locomotion in a two-dimensional fluid flow is described within the finite-dipole model, which consists of a pair of vortices separated by a constant distance. The strength of these vortices is adjusted in real time by the fish to afford orientation and navigation control, in response to of the multi-sensory input from vision, lateral line, and touch. Model parameters for the resulting stochastic differential equations are calibrated through a series of experiments, in which zebrafish swam in a water channel under different illumination conditions. The accuracy of the model is validated through the study of a series of measures of rheotactic behavior, contrasting results of real and in-silico experiments. Our results point at a critical role of hydromechanical feedback during rheotaxis, in the form of a gradient-following strategy.     

'Click' synthesis of a triazole-based inhibitor of Met functions in cancer cells

The use of Cu(I)-catalyzed azide-alkyne 1,3-dipolar cycloaddition permitted the synthesis of a new compound that is able to inhibit the HGF-induced scattering of MDCK (epithelial cells) and in vitro tumorigenesis of H1437 (non-small-cell lung cancer) and GTL-16 (human gastric carcinoma). In agreement with biochemical and biological results, docking studies within the ATP binding site of Met suggested for the new synthesized compound a binding mode similar to that of the active compound Triflorcas previously reported.     

Light as an information carrier underwater

Light is a source of both energy and information for the biota.The spatial, temporal and spectral variability of light experiencedby marine phytoplankton differs significantly from that experiencedby terrestrial plants, due to the selective attenuation of solarirradiance in the aquatic medium. In the present study we analysedsuch variability and focused, in particular, on those bandswithin the spectrum that may act as potential signals for physiologicalresponses. Our results demonstrate that the spectral variationof the light field carries information on the time of day, thevertical position and the presence of very close neighbours,also underwater. This is consistent with the recent findingsof a widespread occurrence of photoreceptors in marine algae.We show also that red photoreceptors, whose presence in marinealgae was difficult to reconcile with the strong attenuationof long wavelengths by water, may be triggered at depth by thered light generated by transpectral processes.     

Superoxide dismutase is preferentially degraded by a proteolytic system from red blood cells following oxidative modification by hydrogen peroxide

The cupro-zinc enzyme superoxide dismutase (SOD) undergoes an irreversible (oxidative) inactivation when exposed to its product, hydrogen peroxide (H2O2). Recent studies have shown that several oxidatively modified proteins (e.g., hemoglobin, albumin, catalase, etc.) are preferentially degraded by a novel proteolytic pathway in the red blood cell. We report that bovine SOD is oxidatively inactivated by exposure to H2O2, and that the inactivated enzyme is selectively degraded by proteolytic enzymes in cell-free extracts of bovine erythrocytes. For example, 95% inactivation of SOD by 1.5 mM H2O2 was accompanied by a 106 fold increase in the proteolytic susceptibility of the enzyme during (a subsequent) incubation with red cell extract. Both SOD inactivation and proteolytic susceptibility increased with H2O2 concentration and/or time of exposure to H2O2. Pre-incubation of red cell extracts with metal chelators, serine reagents, or sulfhydryl reagents inhibited the (subsequent) preferential degradation of H2O2-modified SOD. Furthermore, a slight inhibition of degradation was observed with the addition of ATP. We suggest that H2O2-inactivated SOD is recognized and preferentially degraded by the same. ATP-independent, metallo- serine- and sulfhydryl- proteinase pathway which degrades other oxidatively denatured red cell proteins. Related work in this laboratory suggests that this novel proteolytic pathway may actually consist of a 700 kDa enzyme complex of proteolytic activities. Mature red cells have no capacity for de novo protein synthesis but do have extremely high concentrations of SOD. Red cell SOD generates (and is, therefore, exposed to) H2O2 on a continuous basis, by dismutation of superoxide (from hemoglobin autooxidation and the interaction of hemoglobin with numerous xenobiotics).(ABSTRACT TRUNCATED AT 250 WORDS)