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

Conditions of endoplasmic reticulum stress favor the accumulation of cytosolic prion protein.

After signal sequence-dependent targeting to the endoplasmic reticulum (ER), prion protein (PrP) undergoes several post-translational modifications, including glycosylation, disulfide bond formation, and the addition of a glycosylphosphatidylinositol anchor. As a result, multiple isoforms are generated. Because of the intrinsic weakness of the PrP signal sequence, a fraction of newly synthesized molecules fails to translocate and localizes to the cytosol. The physiopathologic role of this cytosolic isoform is still being debated. Here we have shown that, in both cultured cell lines and primary neurons, ER stress conditions weaken PrP co-translational translocation, favoring accumulation of aggregation-prone cytosolic species, which retain the signal sequence but lack N-glycans and disulfides. Inhibition of proteasomes further increases the levels of cytosolic PrP. Overexpression of spliced XBP1 facilitates ER translocation, suggesting that downstream elements of the Ire1-XBP1 pathway are involved in PrP targeting. These studies reveal a link between ER stress and the formation of cytosolic PrP isoforms potentially endowed with novel signaling or cytotoxic functions

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

State of the Art, Unresolved Issues, and Future Research Directions in the Fight against Hepatitis C Virus: Perspectives for Screening, Diagnostics of Resistances, and Immunization

Hepatitis C virus (HCV) still represents a major public health threat, with a dramatic burden from both epidemiological and clinical points of view. New generation of direct-acting antiviral agents (DAAs) has been recently introduced in clinical practice promising to cure HCV and to overcome the issues related to the interferon-based therapies. However, the emergence of drug resistance and the suboptimal activity of DAAs therapies against diverse HCV genotypes have been observed, determining treatment failure and hampering an effective control of HCV spread worldwide. Moreover, these treatments remain poorly accessible, particularly in low-income countries. Finally, effective screening strategy is crucial to early identifying and treating all HCV chronically infected patients. For all these reasons, even though new drugs may contribute to impacting HCV spread worldwide a preventive HCV vaccine remains a cornerstone in the road to significantly reduce the HCV spread globally, with the ultimate goal of its eradication. Advances in molecular vaccinology, together with a strong financial, political, and societal support, will enable reaching this fundamental success in the coming years. In this comprehensive review, the state of the art about these major topics in the fight against HCV and the future of research in these fields are discussed

Statistical properties and morphology of a 2D gel network at the air/water interface

We report here a detailed characterization of the steps leading to the formation of a bidimensional gel at the air/water interface. The 2D geometry is peculiar in that it allows a continuous and controlled variation of the density of the system over a wide range. The evolution of the statistical properties of the structured network, as a function of the externally controlled density, is related to the corresponding evolution of the mechanical properties. We identified three steps in the gel formation: a first step occurs in the incubation time, in which the individual nm-sized gold nanoparticle aggregate to form quasi 1D structures of typical length of a few microns. The second step occurs in the first stages of the compression, with the growth of the branched structure, finally yielding to the onset of the infinite percolative cluster, which is related to the building of the mechanical elastic modulus. The final step identifies with the homogenization and regularization of the distribution of holes sizes, which relates to the further increase of the elastic modulus upon compression. We speculate that this study should help in the formulation of gels with desired characteristics, such as increased mechanical strength, or increased mobilit

The impact of 10-valent and 13-valent pneumococcal conjugate vaccines on hospitalization for pneumonia in children: A systematic review and meta-analysis

Abstract Background This systematic review and meta-analysis aimed at summarizing available data on the impact of PCV10 and PCV13 in reducing the incidence of CAP hospitalizations in children aged Methods A systematic search of the literature was conducted. We included time-series analyses and before-after studies, reporting the incidence of hospitalization for pneumonia in the periods before and after the introduction of PCV10 or PCV13 into the immunization program. Pooled estimates of Incidence Rate Ratio (IRR) were calculated by using a random-effects meta-analytic model. Results were stratified according to age-groups ( Results A total of 1533 potentially relevant articles were identified. Of these, 12 articles were included in the analysis. In children aged In children aged 24–59 months, the meta-analysis showed a reduction of 9% (95%CI: 5–14%, p-value High heterogeneity was detected among studies evaluating the hospitalization rate for clinically and radiologically confirmed pneumonia. Conclusions The results of this study revealed a significant impact of PCV10 and PCV13 in reducing the hospitalizations for pneumonia, particularly in children age

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