Probing Protein Folding with Sequence-Reversed α-Helical Bundles

Recurrent protein folding motifs include various types of helical bundles formed by α-helices that supercoil around each other. While specific patterns of amino acid residues (heptad repeats) characterize the highly versatile folding motif of four-α-helical bundles, the significance of the polypeptide chain directionality is not sufficiently understood, although it determines sequence patterns, helical dipoles, and other parameters for the folding and oligomerization processes of bundles. To investigate directionality aspects in sequence-structure relationships, we reversed the amino acid sequences of two well-characterized, highly regular four-α-helical bundle proteins and studied the folding, oligomerization, and structural properties of the retro-proteins, using Circular Dichroism Spectroscopy (CD), Size Exclusion Chromatography combined with Multi-Angle Laser Light Scattering (SEC-MALS), and Small Angle X-ray Scattering (SAXS). The comparison of the parent proteins with their retro-counterparts reveals that while the α-helical character of the parents is affected to varying degrees by sequence reversal, the folding states, oligomerization propensities, structural stabilities, and shapes of the new molecules strongly depend on the characteristics of the heptad repeat patterns. The highest similarities between parent and retro-proteins are associated with the presence of uninterrupted heptad patterns in helical bundles sequences.     

Poly[(N‐isopropylacrylamide)‐co‐(itaconic acid)] hydrogels with poly(ethylene glycol)

The aim of the work reported here was to investigate temperature‐ and pH‐sensitive hydrogels of N‐isopropylacrylamide (NIPAM) and itaconic acid (IA) and their semi‐interpenetrating polymer networks (semi‐IPNs) with varying contents of poly(ethylene glycol) (PEG). The stimuli responsiveness, swelling behaviour and mechanical properties of the hydrogels and semi‐IPNs were studied in order to investigate the effect of various amounts of PEG. Pulsed‐gradient spin‐echo NMR experiments were carried out to investigate the diffusion process. The pH sensitivity increased with an increasing amount of PEG in the semi‐IPNs, while the overall rate of water uptake was diffusion‐controlled (n < 0.5). For certain PEG contents (5 and 10 wt%), the semi‐IPNs exhibited better mechanical properties than the poly(NIPAM‐co‐IA) copolymer. The calculated values of the self‐diffusion coefficients of water indicated facilitated diffusion of water through the system with increased amounts of PEG, while the self‐diffusion coefficients of a model compound, metoprolol tartrate, showed no significant dependence on the amount of PEG. According to the results obtained and compared to results reported in the literature, the investigated semi‐IPNs may have potential applications in the controlled release of macromolecular active agents such as proteins and peptides. Copyright © 2009 Society of Chemical Industry     

Synthesis,characterization and application of poly(N-isopropylacrylamide-co-itaconic acid) hydrogels as supports for lipase immobilization

Temperature- and pH-sensitive hydrogels based on N-isopropylacrylamide (NiPAAm) and itaconic acid (IA) were applied for immobilization of lipase from Candida rugosa (CRL). The hydrogels were synthesized by free radical crosslinking copolymerization in the presence of lipase. Characterization of samples by swelling studies, at pH 2.20 and 6.80 at a temperature of 37 °C, scanning electron microscopy (SEM) and Fourier transform infrared analysis (FT-IR) confirmed that the degree of crosslinking, the non-ionic/ionic (NiPAAm/IA) ratio and the enzyme content had impacts on the hydrogel structure, mechanical properties, morphology and swelling kinetics. All hydrogels demonstrated protein loading efficiencies as high as 95 wt.%. A specific activity of the immobilized lipase of around 38 IU/g was attained for an enzyme loading of 20.0 wt.%. As a result, improved pH and temperature optima values were obtained for the immobilized systems in relation to those for the free lipase.     

Visual Change With Moving Displays: More Evidence for Color Feature Map Inhibition During Preview Search.

Preview search with moving stimuli was investigated. The stimuli moved in multiple directions, and preview items could change either their color or their shape before onset of the new (search) displays. In Experiments 1 and 2, the authors found that (a) a preview benefit occurred even when more than 5 moving items had to be ignored, and (b) color change, but not shape change, disrupted preview search in moving stimuli. In contrast, shape change, but not color change, disrupted preview search in static stimuli (Experiments 3 and 4). Results suggest that preview search with moving displays is influenced by inhibition of a color map, whereas preview search with static displays is influenced by inhibition of locations of old distractors. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Energy‐dependent effects of resveratrol in Saccharomyces cerevisiae

The metabolic effects induced by resveratrol have been associated mainly with the consumption of high‐calorie diets; however, its effects with standard or low‐calorie diets remain unclear. To better understand the interactions between resveratrol and cellular energy levels, we used Saccharomyces cerevisiae as a model. Herein it is shown that resveratrol: (a) decreased cell viability in an energy‐dependent manner; (b) lessening of cell viability occurred specifically when cells were under cellular respiration; and (c) inhibition of oxygen consumption in state 4 occurred at low and standard energy levels, whereas at high energy levels oxygen consumption was promoted. These findings indicate that the effects of resveratrol are dependent on the cellular energy status and linked to metabolic respiration. Importantly, our study also revealed that S. cerevisiae is a suitable and useful model to elucidate the molecular targets of resveratrol under different nutritional statuses. Copyright © 2016 John Wiley & Sons, Ltd.     

Connective Tissue Reaction of Rats to a New Zinc‐Oxide‐Eugenol Endodontic Sealer

The aim of this study was to evaluate the biocompatibility in rat subcutaneous connective tissue of a new zinc oxide endodontic sealer (Endomethasone N) compared to those provided by Endofill and Sealer 26. Polyethylene tubes containing the test materials were implanted into dorsal subcutaneous connective tissue of Wistar albino rats. After 7 and 42 days, the implants with the surrounding tissue were collected, fixed, and processed for histologic evaluation. Sections were evaluated for the presence of inflammatory cells (poly or monomorfonuclear), blood vessels, necrosis area, and thickness of fibrous capsule. Comparisons between groups and time‐periods were performed with Kruskal–Wallis and Mann–Whitney U non‐parametric tests for 5% significance level. No differences in the biocompatibility patterns among the materials for the 2 experimental periods were observed. Independently of the sealer, the tissue behavior showed a tendency to decrease the irritation effect over time. It can be concluded that all sealers are irritant, but its toxicity decreased with time. Endomethásone N showed biocompatible characteristics comparable with those provided by Endofill and Sealer 26. Microsc. Res. Tech. 76:1292–1296, 2013. © 2013 Wiley Periodicals, Inc.     

Spatio‐temporal variability in underwater light climate in a turbid river‐floodplain system: Driving factors and estimation using Secchi disc

The underwater light climate has important effects on primary producers. The aim of this research was to evaluate its variability in a turbid river‐floodplain system. Photosynthetically active radiation (PAR) was measured in the Middle Paraná River during different hydrological phases to (a) analyse the photosynthetically active radiation attenuation coefficient (k) and euphotic depth (Z_eu) as well as their associations with optically active components and (b) develop and evaluate indices and regression models based on Secchi disc (SD) measurements to estimate k and Z_eu. Values of k were higher in the fluvial system than in the floodplain and during low‐water stage than high‐water stage. Particulate components controlled the light climate variability. Chromophoric dissolved organic matter and chlorophyll‐a had significant effects during floods. The estimation of k and Z_eu was sensitive to temporal but not to spatial variations. The highest prediction accuracy was observed when using specific non‐linear regressions for each hydrological phase, especially for Z_eu estimation (low stage: k = 1.76 × SD^?0.80, Z_eu = 2.62 × 1/SD^?0.80; high stage: k = 2.04 × SD^?0.53, Z_eu = 2.26 × 1/SD^?0.53). The indices k × SD and Z_eu/SD were significantly different from those proposed for clear water environments. It is concluded that temporal variations should be considered when estimating k and Z_eu in turbid river‐floodplain systems because of the temporal heterogeneity in optically active components. Considering that ecological implication of the light climate depends on Z_eu:depth ratio, we propose to estimate Z_eu instead of k. Finally, indices proposed for clear water environments are not recommended to be applied to turbid environments.