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Abstract To investigate pharmacogenetic interactions among VEGF-A, VEGFR-2, IL-8, HIF-1α, EPAS-1, and TSP-1 SNPs and their role on progression-free survival (PFS) in metastatic breast cancer (MBC) patients treated with bevacizumab plus first-line paclitaxel or with paclitaxel alone. Analyses were performed on germline DNA, and SNPs were investigated by real-time PCR technique. The multifactor dimensionality reduction (MDR) methodology was applied to investigate the interaction between SNPs. The present study was an explorative, ambidirectional cohort study: 307 patients from 11 Oncology Units were evaluated retrospectively from 2009 to 2016, then followed prospectively (NCT01935102). Two hundred and fifteen patients were treated with paclitaxel and bevacizumab, whereas 92 patients with paclitaxel alone. In the bevacizumab plus paclitaxel group, the MDR software provided two pharmacogenetic interaction profiles consisting of the combination between specific VEGF-A rs833061 and VEGFR-2 rs1870377 genotypes. Median PFS for favorable genetic profile was 16.8 vs. the 10.6 months of unfavorable genetic profile (p = 0.0011). Cox proportional hazards model showed an adjusted hazard ratio of 0.64 (95% CI, 0.5–0.9; p = 0.004). Median OS for the favorable genetic profile was 39.6 vs. 28 months of unfavorable genetic profile (p = 0.0103). Cox proportional hazards model revealed an adjusted hazard ratio of 0.71 (95% CI, 0.5–1.01; p = 0.058). In the 92 patients treated with paclitaxel alone, the results showed no effect of the favorable genetic profile, as compared to the unfavorable genetic profile, either on the PFS (p = 0.509) and on the OS (p = 0.732). The pharmacogenetic statistical interaction between VEGF-A rs833061 and VEGFR-2 rs1870377 genotypes may identify a population of bevacizumab-treated patients with a better PFS.

Museum specimens and histologically fixed material are valuable samples for the study of historical soft tissues and represent a possible pathogen-specific source for retrospective molecular investigations. However, current methods for molecular analysis are inherently destructive, posing a dilemma between performing a study with the available technology, thus damaging the sample, and conserving the material for future investigations. Here the authors present the first tests of a non-destructive alternative that facilitates genetic analysis of fixed wet tissues while avoiding tissue damage. The authors extracted DNA from the fixed tissues as well as their embedding fixative solution, to quantify the DNA that was transferred to the liquid component. The results show that human historical DNA can be retrieved from the fixative material of medical specimens and provide new options for sampling valuable collections.

Proton-coupled Oligopeptide Transporters (POTs) of the Major Facilitator Superfamily (MFS) mediate the uptake of short di- and tripeptides in all phyla of life. POTs are thought to constitute the most promiscuous class of MFS transporters, with the potential to transport more than 8400 unique substrates. Over the past two decades, transport assays and biophysical studies have shown that various orthologues and paralogues display differences in substrate selectivity. The E. coli genome codes for four different POTs, known as Di- and tripeptide permeases A-D (DtpA-D). DtpC was shown previously to favor positively charged peptides as substrates. In this study, we describe, how we determined the structure of the 53 kDa DtpC by cryogenic electron microscopy (cryo-EM), and provide structural insights into the ligand specificity of this atypical POT. We collected and analyzed data on the transporter fused to split superfolder GFP (split sfGFP), in complex with a 52 kDa Pro-macrobody and with a 13 kDa nanobody. The latter sample was more stable, rigid and a significant fraction dimeric, allowing us to reconstruct a 3D volume of DtpC at a resolution of 2.7 Å. This work provides a molecular explanation for the selectivity of DtpC, and highlights the value of small and rigid fiducial markers such as nanobodies for structure determination of low molecular weight integral membrane proteins lacking soluble domains.

The In.Te.Se. project - Innovation Territory and Services, for waste management in scattered areas is an Interreg V-A France Italy (ALCOTRA) project, financed within the framework of European cross-border cooperation programmes, in the Alpine region between France and Italy. On the subject of the local exploitation of organic waste, it permits the experimentation of home composting in scattered and cross-border areas in the Italian territories of the Province of Cuneo (Consorzio Servizi Ecologia e Ambiente, CSEA) and in the French areas of the PACA Region (Syndicat Mixte de Traitement des Ordures Ménagères des cantons du Guillestrois et de l'Argentièrois, SMITOMGA), through the use of individual and collective, manual and electromechanical composters. During the project it is estimated that a quantity of organic waste equal to about 2% of the not sorted waste produced in 2019 has been valorised, in 3 municipalities of CSEA and 23 municipalities followed by SMITOMGA where a separate collection circuit does not exist and it is conferred with the general unsorted waste. Overall, 31.72 tons of compost are obtained. The environmental balance deriving from the cooperation of the territories also makes it possible to estimate a negative balance of CO2 produced, with 3212.78 kgCO2 avoided. The economic assessment of not sending the organic component for disposal produced a saving for the two communities as a whole of €10,397.56, involving only 15% of the total population in the municipalities investigated. At the same time, a comparison with a separated collection system for the organic matter, determines the saving of €27,295.73 considering the all tested area. The implementation of this good practice has the potential to achieve interesting results from an environmental, social and economic point of view and to be extended to further portions of the territory and has demonstrated the successful choice of cross-border cooperation and the diversification of applied techniques.

A case study where a tuned mass damper (TMD) was installed at the top of a five-story reinforced concrete (RC) building is presented. The aim of the study was to investigate the effectiveness of the TMD in changing the structural behavior of medium-rise existing buildings from dissipative to non-dissipative in order to eliminate reparation or demolition costs resulting from damages caused by strong earthquakes. The TMD mass is made by a RC slab lying on flat surface sliders. Horizontal stiffness and damping of the TMD are both provided by lead rubber isolators in the first proposed solution and by low-damping rubber isolators and viscous linear dampers, respectively, in the second. The improvement in the building’s structural behavior attained with the installation of the TMD was assessed by considering the flexural demand over capacity ratios of structural elements and the energy dissipated by the TMD. These results are compared with those of the same building retrofitted with a base isolation system. In both of the proposed solutions, TMD remarkably changed the modal behavior of the considered building, improved the flexural verifications, and dissipated most of the input seismic energy. It is therefore demonstrated that a TMD is a valid solution for the retrofit of medium-rise existing buildings.

In the modelling of reinforced concrete (RC) buildings, the rigid diaphragm hypothesis to represent the in-plane behavior of floors was and still is very commonly adopted because of its simplicity and computational cheapness. However, since excessive floor in-plane deformability can cause a very different redistribution of lateral forces on vertical resisting elements, it may be necessary to consider floor deformability. This paper investigates the classical yet intriguing question of modeling orthotropic RC floor systems endowed with lightening elements by means of a uniform orthotropic slab in order to describe accurately the building response under seismic loads. The simplified method, commonly adopted by engineers and based on the equivalence between the transverse stiffness of the RC elements of the real floor and those of the orthotropic slab, is presented. A case study in which this simplified method is used is also provided. Then, an advanced finite element (FE)-based method to determine the elastic properties of the equivalent homogenized orthotropic slab is proposed. The novel aspect of this method is that it takes into account the interaction of shell elements with frame elements in the 3D FE model of the building. Based on the results obtained from the application of this method to a case study, a discussion on the adequacy of the simplified method is also provided.

Viruses can induce the fusion of infected and neighboring cells, leading to the formation of syncytia. Cell–cell fusion is mediated by viral fusion proteins on the plasma membrane of infected cells that interact with cellular receptors on neighboring cells. Viruses use this mechanism to spread rapidly to adjacent cells or escape host immunity. For some viruses, syncytium formation is a hallmark of infection and a known pathogenicity factor. For others, the role of syncytium formation in viral dissemination and pathogenicity remains poorly understood. Human cytomegalovirus (HCMV) is an important cause of morbidity and mortality in transplant patients and the leading cause of congenital infections. Clinical HCMV isolates have broad cell tropism but differ in their ability to induce cell–cell fusions, and little is known about the molecular determinants. We developed a system to analyze HCMV glycoprotein B (gB) variants in a defined genetic background. HCMV strains TB40/E and TR were used as vectors to compare the fusogenicity of six gB variants from congenitally infected fetuses with those from three laboratory strains. Five of them conferred the ability to induce the fusion of MRC-5 human embryonic lung fibroblasts to one or both backbone strains, as determined by a split GFP–luciferase reporter system. The same gB variants were not sufficient to induce syncytia in infected ARPE-19 epithelial cells, suggesting that additional factors are involved. The system described here allows a systematic comparison of the fusogenicity of viral envelope glycoproteins and may help to clarify whether fusion-promoting variants are associated with increased pathogenicity.

The Safer Internet Centre project aims to promote initiatives to make the Internet a better and more secure space for youths. Across the nation, schools were involved in activities aimed at raising awareness throughout a full school year. The first goal of this study is to analyze whether the project was effective, based on three aspects: (a) students’ awareness of online risks; (b) students’ perception of the schools’ policy both on ICTs risk prevention and incidents; and (c) cyberbullying involvement. The second aim is related to the evaluation of the moderating role of intervention dosage on the outcomes. Two trials of intervention were carried out, one for each goal. In the first, we involved students from control schools (N = 675) and experimental schools (N = 775). In the second, we compared two experimental groups: Higher (N = 251, online and face-to-face components) and Lower (N = 315, only the online component) dosage of intervention. Using pre/post-intervention design, we found a significant increase over time in the experimental group on the schools’ commitment to preventing and dealing with online risks. While the students in experimental schools felt themselves significantly more aware of online risks, no significant decrease over time was found in the involvement in cyberbullying. Besides, no dosage effect was found, showing that face-to-face component in our project did not have additional benefits. Implications for future implementations are discussed.

Abstract Novel methods for quantitative, transient-state multiparametric imaging are increasingly being demonstrated for assessment of disease and treatment efficacy. Here, we build on these by assessing the most common Non-Cartesian readout trajectories (2D/3D radials and spirals), demonstrating efficient anti-aliasing with a k-space view-sharing technique, and proposing novel methods for parameter inference with neural networks that incorporate the estimation of proton density. Our results show good agreement with gold standard and phantom references for all readout trajectories at 1.5 T and 3 T. Parameters inferred with the neural network were within 6.58% difference from the parameters inferred with a high-resolution dictionary. Concordance correlation coefficients were above 0.92 and the normalized root mean squared error ranged between 4.2 and 12.7% with respect to gold-standard phantom references for T1 and T2. In vivo acquisitions demonstrate sub-millimetric isotropic resolution in under five minutes with reconstruction and inference times

Abstract The objective of this study was to compare the healing of the augmented sinus at which the antrostomy was covered with a membrane or the repositioned bone plate. Eight sheep underwent bilateral maxillary sinus floor augmentation. The control site was covered with a resorbable membrane, while at the experimental site the bone plate was repositioned, and both were secured with cyanoacrylate. Animals were euthanised after 4 months and histomorphometric analysis was performed. A large amount of the graft appeared to be partially interpenetrated by the newly formed bone. Statistical analysis demonstrated different percentages of the new bone and bone interpenetrated to the graft between test and control site in the close-to-window area respectively 22.1 ± 12.6 vs 7.5 ± 4.5 (P = 0.028) and 66.1 ± 14.7 vs 44.2 ± 15.1 (P = 0.046). Other areas showed no difference in the bone and graft amount. More bone was found at the edges of the antrostomy in the experimental site, without statistical significance. In the centre of the antrostomy, the replaced bony window appeared bonded to the newly formed bone. No remnants and no biological response to cyanoacrylate were observed. The repositioning of the bony window after sinus floor elevation in sheep led to a larger amount of newly formed bone in the close-to-window zone of the grafted area. The bony window appeared partially bonded to the new bone. Newly formed bone was found interpenetrating the graft granules.

Summary: Human cytomegalovirus (HCMV) replicates its DNA genome in specialized replication compartments (RCs) in the host cell nucleus. These membrane-less organelles originate as spherical structures and grow in size over time. However, the mechanism of RC biogenesis has remained understudied. Using live-cell imaging and photo-oligomerization, we show that a central component of RCs, the UL112-113 proteins, undergo liquid-liquid phase separation (LLPS) to form RCs in the nucleus. We show that the self-interacting domain and large intrinsically disordered regions of UL112-113 are required for LLPS. Importantly, viral DNA induces local clustering of these proteins and lowers the threshold for phase separation. The formation of phase-separated compartments around viral genomes is necessary to recruit the viral DNA polymerase for viral genome replication. Thus, HCMV uses its UL112-113 proteins to generate RCs around viral genomes by LLPS to ensure the formation of a pro-replicative environment.

The use of eco-friendly engineered nanomaterials represents a recent solution for an effective and safe treatment of contaminated dredging sludge. In this study, an eco-designed engineered material based on cross-linked nanocellulose (CNS) was applied for the first time to decontaminate a real matrix from heavy metals (namely Zn, Ni, Cu, and Fe) and other undesired elements (mainly Ba and As) in a lab-scale study, with the aim to design a safe solution for the remediation of contaminated matrices. Contaminated freshwater sludge was treated with CNS coupled with a filtering fine-mesh net, and the obtained waters were tested for acute and sublethal toxicity. In order to check the safety of the proposed treatment system, toxicity tests were conducted by exposing the bacterium Aliivibrio fischeri and the crustacean Heterocypris incongruens, while subtoxicity biomarkers such as lysosomal membrane stability, genetic, and chromosomal damage assessment were performed on the freshwater bivalve Dreissena polymorpha. Dredging sludge was found to be genotoxic, and such genotoxicity was mitigated by the combined use of CNS and a filtering fine-mesh net. Chemical analyses confirmed the results by highlighting the abetment of target contaminants, indicating the present model as a promising tool in freshwater sludge nanoremediation.

Despite the widespread use of head-up tilt testing as a means for diagnosing vasovagal syncope, standardization of test methodology remains a controversial issue. The aim of this study was to compare the diagnostic value of head-up tilt testing potentiated with sublingual nitroglycerin with that of head-up tilt testing potentiated with low-dose isoproterenol. For this purpose, 71 patients with unexplained syncope (mean age 43 years) and 30 asymptomatic controls were studied. All underwent the sublingual nitroglycerin and low-dose isoproterenol tests on separate days in a randomized fashion. The protocol of the 2 tests was similar and consisted of 2 phases. Initially, subjects were tilted at 60 degrees for 20 minutes without medication; then, if syncope did not occur, patients and controls received sublingual nitroglycerin (300 microg) or low-dose intravenous isoproterenol (mean infusion rate 1.3 +/- 0.5 microg/min) and continued to be tilted at 60 degrees for a further 20 minutes. During the sublingual nitroglycerin test, a positive response (syncope associated with sudden hypotension and bradycardia) occurred in 35 patients (49%), a negative response in 36 (51%), and drug intolerance in none (0%). During the low-dose isoproterenol test, these percentages were 41%, 59%, and 6%, respectively. A concordant response was observed in 53 cases (75%). Among controls, 3 subjects (10%) had a positive response to the sublingual nitroglycerin test and 4 (13%) to the low-dose isoproterenol test. It is concluded that sublingual nitroglycerin and low-dose isoproterenol are equivalent tests for evaluating patients with unexplained syncope. The sublingual nitroglycerin test, however, is simpler, better tolerated, and safer than the low-dose isoproterenol test and, thus, more suitable for routine clinical use. [ABSTRACT FROM AUTHOR]