Study of cosolvent-induced α-chymotrypsin fibrillogenesis: Does protein surface hydrophobicity trigger early stages of aggregation reaction?

The misfolding of specific proteins is often associated with their assembly into fibrillar aggregates, commonly termed amyloid fibrils. Despite the many efforts expended to characterize amyloid formation in vitro, there is no deep knowledge about the environment (in which aggregation occurs) as well as mechanism of this type of protein aggregation. Alpha-chymotrypsin was recently driven toward amyloid aggregation by the addition of intermediate concentrations of trifluoroethanol. In the present study, approaches such as turbidimetric, thermodynamic, intrinsic fluorescence and quenching studies as well as chemical modification have been successfully used to elucidate the underlying role of hydrophobic interactions (involved in early stages of amyloid formation) in α-chymotrypsin-based experimental system. © 2009 Springer Science+Business Media, LLC

Alpha-2-Macroglobulin Is Acutely Sensitive to Freezing and Lyophilization: Implications for Structural and Functional Studies.

Alpha-2-macroglobulin is an abundant secreted protein that is of particular interest because of its diverse ligand binding profile and multifunctional nature, which includes roles as a protease inhibitor and as a molecular chaperone. The activities of alpha-2-macroglobulin are typically dependent on whether its conformation is native or transformed (i.e. adopts a more compact conformation after interactions with proteases or small nucleophiles), and are also influenced by dissociation of the native alpha-2-macroglobulin tetramer into stable dimers. Alpha-2-macroglobulin is predominately present as the native tetramer in vivo; once purified from human blood plasma, however, alpha-2-macroglobulin can undergo a number of conformational changes during storage, including transformation, aggregation or dissociation. We demonstrate that, particularly in the presence of sodium chloride or amine containing compounds, freezing and/or lyophilization of alpha-2-macroglobulin induces conformational changes with functional consequences. These conformational changes in alpha-2-macroglobulin are not always detected by standard native polyacrylamide gel electrophoresis, but can be measured using bisANS fluorescence assays. Increased surface hydrophobicity of alpha-2-macroglobulin, as assessed by bisANS fluorescence measurements, is accompanied by (i) reduced trypsin binding activity, (ii) increased chaperone activity, and (iii) increased binding to the surfaces of SH-SY5Y neurons, in part, via lipoprotein receptors. We show that sucrose (but not glycine) effectively protects native alpha-2-macroglobulin from denaturation during freezing and/or lyophilization, thereby providing a reproducible method for the handling and long-term storage of this protein.Early Career Fellowship from the National Health and Medical Research Council GNT1012521(A.R.W.); Wellcome Trust Programme Grant (J.R.K., C.M.D.) 094425/Z/10/Z; Samsung GRO Grant (M.R.W.)This is the final version of the article. It first appeared from PLoS via http://dx.doi.org/10.1371/journal.pone.013003

Strategic crossing of biomass and harvest index—source and sink—achieves genetic gains in wheat

To accelerate genetic gains in breeding, physiological trait (PT) characterization of candidate parents can help make more strategic crosses, increasing the probability of accumulating favorable alleles compared to crossing relatively uncharacterized lines. In this study, crosses were designed to complement “source” with “sink” traits, where at least one parent was selected for favorable expression of biomass and/or radiation use efficiency—source—and the other for sink-related traits like harvest-index, kernel weight and grains per spike. Female parents were selected from among genetic resources—including landraces and products of wide-crossing (i.e. synthetic wheat)—that had been evaluated in Mexico at high yield potential or under heat stress, while elite lines were used as males. Progeny of crosses were advanced to the F4 generation within Mexico, and F4-derived F5 and F6 generations were yield tested to populate four international nurseries, targeted to high yield environments (2nd and 3rd WYCYT) for yield potential, and heat stressed environments (2nd and 4th SATYN) for climate resilience, respectively. Each nursery was grown as multi-location yield trials. Genetic gains were achieved in both temperate and hot environments, with most new PT-derived lines expressing superior yield and biomass compared to local checks at almost all international sites. Furthermore, the tendency across all four nurseries indicated either the superiority of the best new PT lines compared with the CIMMYT elite checks, or the superiority of all new PT lines as a group compared with all checks, and in some cases, both. Results support—in a realistic breeding context—the hypothesis that yield and radiation use efficiency can be increased by improving source:sink balance, and validate the feasibility of incorporating exotic germplasm into mainstream breeding efforts to accelerate genetic gains for yield potential and climate resilience

Evaluation of CIMMYT synthetic hexaploid wheats for resistance to septoria tritici blotch

Of 128 genotypes, 60 synthetic hexaploid wheat genotypes showed no resistance response, whereas the remaining genotypes showed specific resistance to one or more isolates. Interestingly, nine genotypes were resistant to all isolates tested. Isolate RM46 collected from Khuzestan Province was virulent on 87% of SHWs, suggesting that it has the lowest number of avirulence genes. Isolate RM155 collected from Golestan Province displayed the highest number of incompatible interactions (n=42), indicating that it possesses the highest number of avirulence genes. RM151 was the most aggressive isolate with the highest mean disease severity (69%), whereas RM41 was the least aggressive isolate with the lowest mean disease severity (37%). The present study was the first conducted to evaluate CIMMYT SHWs for resistance to Iranian Z. tritici isolates. Our results showed that some SHWs possess a broad spectrum of resistance gene(s) or a combination of a set of effective genes against various STB isolates

Comparison between Removal Efficiency of Slag, zeolite, and Conventional media in slow sand Filter for Removal of Lead and Cadmium from Water Resources

Introduction: Heavy metals owing to their health hazards and high toxicity in low concentration for human and environment have very concern and attention. Slow sand filter is one of the simple and cost-effective for removal of these pollutants. In this method, media play an important role for removal of pollutant. Therefore, the aim of this study was investigation of different media like slag, zeolite, and conventional media in slow sand filter for removal of lead and cadmium. Methods: In this research there are three beds filter include typical filter bed, slag and zeolite that used in pilot plant for investigation of lead and cadmium removal at three concentration of 0.1T 1 and 10 ppm. Each of filters has an internal diameter of 8 cm and a height of 120 cm with Plexiglas, which have a continuous flow operation. Results: The removal efficiency of turbidity by three typical filter bed, slag, and zeolite with initial turbidity of 13 NTU was 46%, 77%, and 89% respectively. Removal efficiency of lead without turbidity was 70.3%, 79%, and 59.8% respectively for 0.1 ppm lead. For 1 ppm, concentration of lead removal efficiency was 51.8%, 52.7% and 52.6% respectively and for 10 ppm it was 53.4%, 57.8%, and 59.8% respectively. Cadmium removal for these media was 23.4%, 37.5%, and 59.4% respectively at 0.1 ppm cadmium. At 1 ppm of cadmium concentration, it was 37.9%, 45% and 41.3% respectively and at 10 ppm concentration of cadmium it was 68.3%, 68.6% and 67% respectively. Conclusion: Slag and zeolite beds are more efficiently than the conventional sand beds in the slow sand filter, so it can be used instead of the usual sand for removing lead and cadmium from resources water

Peptides NAP and SAL attenuate human tau granular-shaped oligomers in vitro and in SH-SY5Y cells

Accumulation of human tau protein in the central nervous system is an outstanding feature of Alzheimer's Diseases and other tauopathies. Among the aggregate species, granular-shaped oligomers are recently reported as toxic species and associated with neuronal loss. Many interests have been focused on the reducing of these neurotoxic oligomers in tauopathies. It has been approved that peptides NAP (NAPVSIPQ) and SAL (SALLRSIPA), snipped from activity-dependent neurotrophic protein (ADNP) and activity-dependent neurotrophic factor (ADNF), protect brain against a wide range of toxins in neurodegenerative models. The neuroprotection mechanisms of NAP and SAL are under investigation. We report here that the aggregation pathway of human tau protein, in the presence of NAP or SAL, could be affected and moved toward the less granular-shaped species. Fluorophore binding assays, kinetic parameters and circular dichroism records, showed the presence of unusual aggregated species upon treatment with NAP or SAL, as well as hydrophobicity studies. Also, sizing and morphological investigation of aggregates, using dynamic scattered light and Atomic Force Microscopy, elucidated a significant reduction of granular-shaped oligomers under 50 nm. Furthermore, immunospecific detection of oligomers in SH-SY5Y cell line, using flow cytometric assessment supported our results. We conclude that one of the possible mechanisms of neuronal protection by NAP or SAL could be the attenuation of granular-shaped oligomeric tau under stress condition. © 201