Lysozyme inactivation and aggregation in stirred-reactor

Mechanisms of enzyme inactivation and aggregation are still poorly understood. In this work, we are considering the characterisation of both inactivation and aggregation in stirred tank reactor, with lysozyme as the model enzyme.

The inactivation kinetics are first order. For stirring speeds in the range of 0–700 rpm, the kinetic constant is found to be proportional to the power brought by the impeller. It suggests that inactivation depends on collisions between enzyme molecules. Efficient collisions between native and inactive molecules induce native molecules to turn into inactive molecules and lead to lysozyme aggregation.

During inactivation, enzymes are found to aggregate as shown by light scattering measurements. The structure of aggregates was studied on samples treated for chemical denaturation and reduction. The aggregates are supramolecular edifices, mainly made up of inactivated enzymes linked by weak forces. But aggregates are also made up of dimers and trimers of lysozyme, linked by disulfide bridges. Dimers and trimers are 18% and 5%, respectively, of the total amount of lysozyme aggregates.

Whatever the stage of aggregate formation and the initial enzyme concentration are, these aggregates are irreversibly inactivated. Enzyme activity is definitely lost even if stirring is stopped and/or temperature decreased.

This study points out the importance of hydrodynamics in bioreactors and highlights the nature of the aggregates resulting from the interactions between native and inactive enzymes.     

Scale specific determinants of tree diversity in an old growth temperate forest in China

The major processes generating pattern in plant community composition depend upon the spatial scale and resolution of observation, therefore understanding the role played by spatial scale on species patterns is of major concern. In this study, we investigate how well environmental (topography and soil variables) and spatial variables explain variation in species composition in a 25-ha temperate forest in northeastern China. We used new variation partitioning approaches to discover the spatial scale (using multi-scale spatial PCNM variables) at which environmental heterogeneity and other spatially structured processes influence tree community composition. We also test the effect of changing grain of the study (i.e. quadrat size) on the variation partitioning results. Our results indicate that (1) species composition in the Changbai mixed broadleaf-conifer forest was controlled mainly by spatially structured soil variation at broad scales, while at finer scales most of the explained variation was of a spatial nature, pointing to the importance of biotic processes. (2) These results held at all sampling grains. However, reducing quadrat size progressively reduced both spatially and environmentally explained variance. This probably partly reflects increasing stochasticity in species abundances, and the smaller proportion of quadrats occupied by each species, when quadrat size is reduced. The results suggest that environmental heterogeneity (i.e. niche processes) and other biotic processes such as dispersal work together, but at different spatial scales, to build up diversity patterns.     

Integrating ecological features of species in spatial pattern analysis of a plant community

Objective: This empirical study was designed to explore the role of ecological features of species in the spatial patterning of a grassland community. Location: Banks of the river Rhône in France. Material and Methods: First, we explored the spatial pattern of 29 species recorded in the community using spatial autocorrelation analysis of species cover values. Second, we then explored the relationship between the patterns found and a set of life attributes that characterized the ecological features of species for resource foraging or dispersion. Finally, we explored the spatial relationship of groups of species that shared the same ecological features using cross‐correlation analysis. Results: We found a significant relationship between the spatial pattern and life attributes of the species highlighting three groups of species: (1) species characterized as competitors, reproducing by runner clonal organs and forming large, dense patches; (2) species characterized as competitive‐rud‐erals, dispersing exclusively by seed production and forming small periodic patches; and (3) species classified as CSR, characterized by rosette morphology and short rhizomes as clonal organs without any significant spatial autocorrelation. Spatial segregation was found between group 1 and group 2 up to 14 m; no significant cross‐correlation between groups 1 and 3 between 0 and 3.5 m, and association between groups 2 and 3 up to 14 m. Conclusions: These results helped to understand how species attributes (relative to stature or dispersion abilities); external factors (such as disturbance) and biotic processes (competition) interplay in structuring the plant community under study in space.     

小鞘指环虫种群的聚集性研究

本文中建立了一个衡量聚集性的指标－聚集度Ａ,并用该指标结合方差／均数和负二项分布的参数ｋ值分析了小鞘指环虫（Ｄａｃｔｙｌｏｇｙｒｕｓｖａｇｉｎｕｌａｔｕｓ）种群在其宿主鲢（Ｈｙｐｏｐｈｔｈａｌｍｉｃｈｔｈｙｓｍｏｌｉｔｒｉｘ）种群中分布的聚集程度与感染率和丰度的关系。结果表明该虫种群的聚集程度随感染率及丰度的上升而呈现下降的特点,作为衡量聚集性的指标,聚集度要优于ｋ值,而后者又优于方差与均数之比,     

The Glycine-Alanine Dipeptide Repeat from C9orf72 Hexanucleotide Expansions Forms Toxic Amyloids Possessing Cell-to-Cell Transmission Properties

Hexanucleotide expansions, GGGGCC, in the non-coding regions of the C9orf72 gene were found in major frontotemporal lobar dementia and amyotrophic lateral sclerosis patients (C9FTD/ALS). In addition to possible RNA toxicity, several dipeptide repeats (DPRs) are translated through repeat-associated non-ATG-initiated translation. The DPRs, including poly(GA), poly(GR), poly(GP), poly(PR), and poly(PA), were found in the brains and spinal cords of C9FTD/ALS patients. Among the DPRs, poly(GA) is highly susceptible to form cytoplasmic inclusions, which is a characteristic of C9FTD/ALS. To elucidate DPR aggregation, we used synthetic (GA)₁₅ DPR as a model system to examine the aggregation and structural properties in vitro. We found that (GA)₁₅ with 15 repeats fibrillates rapidly and ultimately forms flat, ribbon-type fibrils evidenced by transmission electron microscopy and atomic force microscopy. The fibrils are capable of amyloid dye binding and contain a characteristic cross-β sheet structure, as revealed by x-ray scattering. Furthermore, using neuroblastoma cells, we demonstrated the neurotoxicity and cell-to-cell transmission property of (GA)₁₅ DPR. Overall, our results show the structural and toxicity properties of GA DPR to facilitate future DPR-related therapeutic development.     

The role of nurse plants in the establishment of shrub seedlings in the semi-arid subtropical Andes

In the nurse plant syndrome, or nurse association, seedlings (beneficiaries) are associated with adult shrubs/trees (benefactors). This phenomenon has been documented in several regions of the planet. Abiotic stress amelioration (one mechanism of facilitation) is one of the causes of this association. Most of the studies addressing the nurse syndrome have been conducted on spatial scales of a few hectares and have focused on only one or a few species. Moreover, there is an almost complete lack of studies addressing the incidence and characteristics of the nurse phenomenon in the arid Andes of South America. We undertook a first approximation to the study of facilitation in these ecosystems. The study was conducted at local and regional scales and involved the assessment of the spatial distribution of juveniles (seedlings and saplings) of 51 populations of 16 shrub and 12 cactus species in relation to shrub cover at 20 localities of the Prepuna (subtropical Andes of Bolivia and Argentina, 20–26°S). In terms of spatial distribution, the juveniles of most of the populations of shrubs studied were distributed both under the shrubs and in open spaces, thereby showing an apparent indifference to microhabitat. Globose and opuntioid cacti were preferentially distributed below the canopies of shrubs and were usually more associated with the dominant shrub species, which stood out as better potential nurses. The pattern was consistent throughout the region, including the more mesic and arid localities. The fact that Prepuna woody species are capable of establishing in open spaces would confer this region a greater resilience. Our findings further suggest that community dynamics in arid and semi-arid environments are more variable than previously thought.     

Community pattern analysis: A method for quantifying community mosaic structure

A method of quantifying community spatial patterns, community pattern analysis, is described. It is proposed that ordination analysis is used to obtain an integrated score for each quadrat from transect data. For the data presented here, separate ordinations were made of both floristic and environmental (soils) data. The ordination axis scores are then analysed using two or three-term local variance analysis to quantify the scales of community pattern. Correlation analyses allow the relationship between the vegetation and soils data (as represented by ordination axis scores), and other environmental data to be investigated at defined scales. The advantages of this method, that employs the joint application of conventional methods, are that it includes the influence of all species in the analysis, and that multiple uncorrelated scales of pattern within a community are identified.     

Monte Carlo-based assessment of the trade-off between spatial resolution,field-of-view and scattered radiation in the variable resolution X-ray CT scanner

ObjectiveThe purpose of this work is to evaluate the impact of optimization of magnification on performance parameters of the variable resolution X-ray (VRX) CT scanner.MethodsA realistic model based on an actual VRX CT scanner was implemented in the GATE Monte Carlo simulation platform. To evaluate the influence of system magnification, spatial resolution, field-of-view (FOV) and scatter-to-primary ratio of the scanner were estimated for both fixed and optimum object magnification at each detector rotation angle. Comparison and inference between these performance parameters were performed angle by angle to determine appropriate object position at each opening half angle.ResultsOptimization of magnification resulted in a trade-off between spatial resolution and FOV of the scanner at opening half angles of 90°–12°, where the spatial resolution increased up to 50% and the scatter-to-primary ratio decreased from 4.8% to 3.8% at a detector angle of about 90° for the same FOV and X-ray energy spectrum. The disadvantage of magnification optimization at these angles is the significant reduction of the FOV (up to 50%). Moreover, magnification optimization was definitely beneficial for opening half angles below 12° improving the spatial resolution from 7.5 cy/mm to 20 cy/mm. Meanwhile, the FOV increased by more than 50% at these angles.ConclusionIt can be concluded that optimization of magnification is essential for opening half angles below 12°. For opening half angles between 90° and 12°, the VRX CT scanner magnification should be set according to the desired spatial resolution and FOV.     

Helianthus maximiliani and species fine‐scale spatial pattern affect diversity interactions in reconstructed tallgrass prairies

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