Role of Individual Disulfide Bonds in the Structural Maturation of a Low Molecular Weight Glutenin Subunit

Gliadins and glutenins are the major storage proteins that accumulate in wheat endosperm cells during seed development. Although gliadins are mainly monomeric, glutenins consist of very large disulfide-linked polymers made up of high molecular weight and low molecular weight subunits. These polymers are among the largest protein molecules known in nature and are the most important determinants of the viscoelastic properties of gluten. As a first step toward the elucidation of the folding and assembly pathways that lead to glutenin polymer formation, we have exploited an in vitro system composed of wheat germ extract and bean microsomes to examine the role of disulfide bonds in the structural maturation of a low molecular weight glutenin subunit. When conditions allowing the formation of disulfide bonds were established, the in vitro synthesized low molecular weight glutenin subunit was recovered in monomeric form containing intrachain disulfide bonds. Conversely, synthesis under conditions that did not favor the formation of disulfide bonds led to the production of large aggregates from which the polypeptides could not be rescued by the post-translational generation of a more oxidizing environment. These results indicate that disulfide bond formation is essential for the conformational maturation of the low molecular weight glutenin subunit and suggest that early folding steps may play an important role in this process, allowing the timely pairing of critical cysteine residues. To determine which cysteines were important to maintain the protein in monomeric form, we prepared a set of mutants containing selected cysteine to serine substitutions. Our results show that two conserved cysteine residues form a critical disulfide bond that is essential in preventing the exposure of adhesive domains and the consequent formation of aberrant aggregates

Opening and closure forces of sliding mechanisms of different self-ligating brackets

Self-ligating brackets engage the wire by means of a slide mechanism. Forces that have to be applied to open and close the sliding mechanism of brackets are still unknown. Objective The aim of this study was to measure and compare the opening and closure forces of different self-ligating brackets. Material and Methods Three different stainless steel self-ligating brackets (Carriere LX, Ortho Organizers; F1000, Leone; Damon Q, Ormco) were tested. For each different bracket, 20 maxillary right central incisors and 20 mandibular right central incisors were used. Opening and closure forces were measured using an Instron Universal Testing Machine. Statistical analysis was performed and ANOVA and Tukey tests were carried out. Results Opening forces were registered between 1.1 N and 5.6 N, whereas closure forces were recorded between 1.57 N and 4.87 N. Significant differences were detected among the different brackets and between the two prescriptions tested. Conclusion The knowledge of different opening and closure forces of self-ligating brackets can help the orthodontist in the clinical management of these devices

Impaired glucose metabolism in subjects with the Williams-Beuren syndrome. A five-year follow-up cohort study

Objective. The Williams-Beuren syndrome (WS) is associated with impaired glucose metabolism (IGM) early in adulthood. However, the pathophysiology of IGM remains poorly defined, due to the lack longitudinal studies investigating the contribution of β-cell dysfunction and impaired insulin sensitivity. This study aimed at assessing incidence of IGM and the underlying mechanisms in WS adults. Methods. This observational, longitudinal (5-year), cohort study enrolled thirty-one consecutive WS subjects attending a tertiary referral center. An oral glucose tolerance test (OGTT) was performed yearly and used to classify patients as normal or IGM, including impaired fasting glucose (IFG) and/or impaired glucose tolerance (IGT) and diabetes mellitus (DM), and to calculate surrogate measures of insulin secretion and/or sensitivity. Results. IGM patients were 18 (58.1%, three DM) at baseline and 19 (61.3%, five DM) at end-of-follow-up. However, 13 individuals changed category of glucose homeostasis in both directions during follow-up (8 progressors, 5 regressors) and 18 did not (8 non-progressors, 10 non-regressors). New cases of IGM and DM were 11.1 and 2.53 per 100 persons-year, respectively, and were treated non-pharmacologically. In the whole cohort and, to a higher extent, in progressors, indices of early-phase insulin secretion and insulin sensitivity decreased significantly from baseline to end-of-follow-up, with concurrent reduction of the oral disposition index and insulin secretion-sensitivity index-2 (ISSI-2), compensating insulin secretion for the level of insulin resistance. No baseline measure independently predicted progression, which correlated with change from baseline in ISSI-2. Compared with patients with normal glucose homeostasis, IGT subjects had impaired insulin sensitivity, whereas insulin secretion was reduced only in those with IFG+IGT or DM. Conclusions. IGM incidence is high in young adults with WS, suggesting the need of early screening and timed intervention. As in classical type 2 diabetes, impaired insulin sensitivity and β-cell dysfunction contribute, in this sequence, to progression to IGM and DM

A new 3D concentration gradient maker and its application in building hydrogels with a 3D stiffness gradient

For a deeper knowledge of phenomena at cell and tissue level, for understanding the role on bimolecular signalling and for the development of new drugs it is important to recreate in vitro environments that mimic the physiological one. Spatial gradients of soluble species guide the cells' morphogenesis, and they range in a three-dimensional (3D) environment. Gradients of mechanical properties, which have a 3D pattern, could lead cell migration and differentiation. In this work, a new 3D Concentration Gradient Maker able to generate 3D concentration gradients of soluble species was developed, which could be used for differential perfusion of scaffolds. The same device can be applied to build hydrogel matrixes with a 3D gradient of mechanical properties. Computational dynamic fluid analysis was used to develop the gradient generator; the validation of the 3D gradient of stiffness was carried out using finite elements analysis and experimental studies. The device and its application could bring improvements in studying phenomena related to cell chemotaxis and mechanotaxis, but also to differentiation in the simultaneous presence of gradients in both soluble chemical species and substrate stiffnes

Genipin diffusion and reaction into a gelatin matrix for tissue engineering applications

Genipin is a natural low-toxic cross-linker for molecules with primary amino groups, and its use with collagen and gelatin has shown a great potential in tissue engineering applications. The fabrication of scaffolds with a well-organized micro and macro topology using additive manufacturing systems requires an accurate control of working parameters, such as reaction rate, gelling time, and diffusion constant. A polymeric system of 5% w/v gelatin in PBS with 2 mg/mL collagen solutions in a 1:1 weight ratio was used as template to perform measurements varying genipin concentration in a range of 0.1-1.5% w/w with respect to gelatin. In the first part of this work, the reaction rate of the polymeric system was estimated using a new colorimetric analysis of the reaction. Then its workability time, closely related to the gelling time, was evaluated thanks to rheological analysis: finally, the quantification of static and dynamic diffusion constants of genipin across nonreacting and reacting membranes, made respectively by agarose and gelatin, was performed. It was shown that the colorimetric analysis is a good indicator of the reaction progress. The gelling time depends on the genipin concentration, but a workability window of 40 min guaranteed up to 0.5% w/w genipin. The dynamic diffusion constant of genipin in the proposed polymeric system is in the order of magnitude of 10(-7) . The obtained results indicated the possibility to use the genipin, gelatin, and collagen, in the proposed concentrations, to build well-defined hydrogel scaffolds with both extrusion-based and 3D ink-jet system. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2015

Dictionnaire des biens communs

Dictionnaire des biens communs. © Presses universitaires de France. C’est en effet intéressant de revenir à la genèse de ce dictionnaire. L’idée en est venue alors que nous lancions en 2010, à plusieurs, économistes, juristes et historiens, un programme financé par l’Agence nationale de la recherche (ANR), sur le thème « propriété intellectuelle, communs et exclusivité » (« Propice »). Ce projet avait pour objet de réfléchir sur ce que certains ont dénommé les « communs de la connaissance » e..

Data-driven Predictive Latency for 5G: A Theoretical and Experimental Analysis Using Network Measurements

The advent of novel 5G services and applications with binding latency requirements and guaranteed Quality of Service (QoS) hastened the need to incorporate autonomous and proactive decision-making in network management procedures. The objective of our study is to provide a thorough analysis of predictive latency within 5G networks by utilizing real-world network data that is accessible to mobile network operators (MNOs). In particular, (i) we present an analytical formulation of the user-plane latency as a Hypoexponential distribution, which is validated by means of a comparative analysis with empirical measurements, and (ii) we conduct experimental results of probabilistic regression, anomaly detection, and predictive forecasting leveraging on emerging domains in Machine Learning (ML), such as Bayesian Learning (BL) and Machine Learning on Graphs (GML). We test our predictive framework using data gathered from scenarios of vehicular mobility, dense-urban traffic, and social gathering events. Our results provide valuable insights into the efficacy of predictive algorithms in practical applications

Impact of Candida albicans hyphal wall protein 1 (HWP1) genotype on biofilm production and fungal susceptibility to microglial cells

The hyphal wall protein 1 (HWP1) gene of Candida albicans encodes for a fungal cell wall protein, required for hyphal development and yeast adhesion to epithelial cells; yet, its role in pathogenesis remains largely unknown. In the present study, we analyzed two C. albicans laboratory strains, the DAY286 (HWP1/HWP1) and the null mutant FJS24 (hwp1/hwp1) and six clinical isolates [3 harbouring the homozygous HWP1 gene (HWP1/HWP1) and 3 the heterologous gene (HWP1/hwp1)]. Biofilm production, fungal HWP1 mRNA levels and ultrastructural morphology were investigated; also, the susceptibility of these strains to microglial cells was evaluated, in terms of fungal damage and immune cell-mediated secretory response. When comparing the two laboratory strains, biofilm was produced to a similar extent independently on the genetic background, while the susceptibility to microglial cell-mediated damage was higher in the hwp1/hwp1 mutant than in the HWP1/HWP1 counterpart. Also, transmission electron microscopy revealed differences between the two in terms of abundance in surface adhesin-like structures, fungal cell wall shape and intracellular granules. When comparing the clinical isolates grouped according to their HWP1 genotype, reduced biofilm production and increased susceptibility to microglial cell-mediated damage occurred in the HWP1/hwp1 isolates with respect to the HWP1/HWP1 counterparts; furthermore, upon exposure to microglial cells, the HWP1/HWP1 isolates, but not the HWP1/hwp1 counterpart, showed enhanced HWP1 mRNA levels. Finally, both laboratory and clinical isolates exhibited reduced ability to stimulate TNFα and nitric oxide production by microglial cells in the case of heterozygous or null mutant HWP1 genotype. Overall, these data indicate that C. albicans HWP1 genotype influences pathogen morphological structure as well as its interaction with microglial cells, while fungal biofilm production results unaffected, thus arguing on its role as virulence factor that directly affects host mediated defences

A cross-sectional study investigating lifestyle and weight perception of undergraduate students in southern Italy

BACKGROUND: The aim of the study was to explore lifestyle of undergraduate students in southern Italy and to investigate their determinants. METHODS: An anonymous, web-based questionnaire investigating weight and its perception, smoking and alcohol use, achievement of recommended levels of physical activity and time spent daily in screen-based sedentary behaviors was administered to students attending two universities in southern Italy. Age, gender, city, type of degree course attended, occupational status and residential status were considered as possible demographic determinants. RESULTS: The majority of the participants reported a normal weight (71.2%), were non-smokers (66.6%), occasional alcohol consumers (60.5%) and insufficiently active (62.6%), with a reported mean screen time exceeding 2 h per day. Gender, city, type of degree course and occupational status were found to be associated with lifestyle by regression analysis. Normal weight, overweight and obese students were generally properly aware of their weight condition; however, weight misperceptions were registered among normal and underweight undergraduates. CONCLUSIONS: This study highlighted some critical issues regarding lifestyles of university students that suggest the need for health promotion interventions targeted mainly on physical activity

Engineering random spin models with atoms in a high-finesse cavity

All-to-all interacting, disordered quantum many-body models have a wide range of applications across disciplines, from spin glasses in condensed-matter physics, over holographic duality in high-energy physics, to annealing algorithms in quantum computing. Typically, these models are abstractions that do not find unambiguous physical realisations in nature. Here, we realise an all-to-all interacting, disordered spin system by subjecting an atomic cloud in a cavity to a controllable light shift. Adjusting the detuning between atom resonance and cavity mode, we can tune between disordered versions of a central-mode model and a Lipkin-Meshkov-Glick model. By spectroscopically probing the low-energy excitations of the system, we explore the competition of interactions with disorder across a broad parameter range. We show how disorder in the central-mode model breaks the strong collective coupling, making the dark state manifold cross over to a random distribution of weakly-mixed light-matter, "grey", states. In the Lipkin-Meshkov-Glick model the ferromagnetic finite-size ground state evolves towards a paramagnet as disorder is increased. In that regime, semi-localised eigenstates emerge, as we observe by extracting bounds on the participation ratio. These results present significant steps towards freely programmable cavity-mediated interactions for the design of arbitrary spin Hamiltonians.Comment: 8 pages, 4 figures, methods, supplementary materia