Evaluation of the Molecular Landscape in PD-L1 Positive Metastatic NSCLC: Data from Campania,Italy

Background: Immune-checkpoint inhibitors (ICIs) have increased and improved the treatment options for patients with non-oncogene-addicted advanced stage non-small cell lung cancer (NSCLC). However, the role of ICIs in oncogene-addicted advanced stage NSCLC patients is still debated. In this study, in an attempt to fill in the informational gap on the effect of ICIs on other driver mutations, we set out to provide a molecular landscape of clinically relevant oncogenic drivers in programmed death-ligand 1 (PD-L1) positive NSCLC patients. Methods: We retrospectively reviewed data on 167 advanced stage NSCLC PD-L1 positive patients (≥1%) who were referred to our clinic for molecular evaluation of five driver oncogenes, namely, EGFR, KRAS, BRAF, ALK and ROS1. Results: Interestingly, n = 93 (55.7%) patients showed at least one genomic alteration within the tested genes. Furthermore, analyzing a subset of patients with PD-L1 tumor proportion score (TPS) ≥ 50% and concomitant gene alterations (n = 8), we found that n = 3 (37.5%) of these patients feature clinical benefit with ICIs administration, despite the presence of a concomitant KRAS gene alteration. Conclusions: In this study, we provide a molecular landscape of clinically relevant biomarkers in NSCLC PD-L1 positive patients, along with data evidencing the clinical benefit of ICIs in patient NSCLC PD-L1 positive alterations.     

A model for representing topological relationships between complex geometric features in spatial databases

Various models for the representation of topological relationships have been developed. The aim of this paper is to show that the set of relationships proposed in [7] (the CBM), for describing topological relationships among two-dimensional simple features, is applicable with few modifications to the case of complex features (that is, areas made up of several components possibly containing holes, lines with self-intersections, and/or more than two endpoints, and so on). The CBM offers a small set of topological relationships with high expressiveness which is proven to be mutually exclusive and complete, and therefore suitable to be embedded in a spatial query language.     

Wall effects for the pressure drop in fixed beds

A very simple model is presented for the effect of the container wall on the pressure drop in a fluid flowing through a bed of spheres. It is based on the application of the Ergun equation to the bulk region of the bed, unaffected by the wall. Pressure loss predictions are found to correspond well to the opposing trends reported in the literature for the viscous- and inertial-flow regimes.     

A review of experimental investigations and assessment methods for masonry arch bridges

Masonry arch bridges constitute a significant proportion of European road and rail infrastructures. Most of them are well over 100 years old and are supporting traffic loads many times above those originally envisaged. The inherent variation in their constituent materials, the traditional design criteria and methods used for their construction, their deterioration over time caused by weathering processes and the development of other defects, significantly influence the mechanical response of these historic structures. A deep understanding on the numerous factors that affect the structural behaviour of masonry arch bridges and on the analysis methods to assess the life expectancy of such bridges and inform maintenance and strengthening strategies is essential. This paper provides a critical review of the experimental studies that have been carried out and of the assessment approaches that have been developed in the last three decades to these aims. The current knowledge is established and areas of possible future research work are identified, with the aim of providing students and researchers, asset managers and bridge owners, and practitioners with a guidance for research activities and maintenace strategies.     

Electronic coupling between azurin and gold at different protein/substrate orientations

By means of constrained classical molecular dynamics simulations, we have computed the structure of azurin deposited on a Au(111) surface at different possible orientations and the azimuthal forces acting on the protein at each sampled conformation. We have then evaluated the effect of the angular variation on the speed of electron tunneling between the protein redox site and the metal surface. We find that the azurin/gold electronic coupling has a strong dependence on the molecular orientation and is greatly enhanced by inclining the protein to lie as flat as possible on the surface. We discuss the implications of our results for scanning probe microscopy experiments in which tunneling currents are measured while the protein is subjected to mechanical forces exerted by the tip of the instrument.     

Iron Chelates: Production Processes and Reaction Evolution Analysis

Nowadays, fertilization using synthetic chelates is the most common technique to address iron chlorosis, a disease that affects plant growth. Ethylenediamine-N,N'-bis(o-hydroxyphenyl) acetic acid (EDDHA) is among the most efficient iron chelating agents. To produce EDDHA, a reaction was performed using as reactants phenol, ethylenediamine, glyoxylic acid, and sodium hydroxide. To study the reaction kinetics, samples were withdrawn from the reactor during the reaction and the kinetics was quantified, evaluating the yield evolution during the reaction phase. This study was useful to optimize the reaction time. Then, a catalyst was added to the reaction mixture to analyze its effect on the reaction evolution. Comparing the reaction evolution of the non-catalyzed and the catalyzed reaction protocols, two main results have to be highlighted: the time to reach the final yield is lower than the one proposed in the literature and the used catalyst has a minimum effect on the reaction rate.     

Congenital diarrheal disorders: an updated diagnostic approach

Congenital diarrheal disorders (CDDs) are a group of inherited enteropathies with a typical onset early in the life. Infants with these disorders have frequently chronic diarrhea of sufficient severity to require parenteral nutrition. For most CDDs the disease-gene is known and molecular analysis may contribute to an unequivocal diagnosis. We review CDDs on the basis of the genetic defect, focusing on the significant contribution of molecular analysis in the complex, multistep diagnostic work-up.     

Environmental factors affect the activity of biocontrol agents against ochratoxigenic Aspergillus carbonarius on wine grape

The influence of temperature and relative humidity (RH) on the activity of three biocontrol agents-the yeast Metschnikowia pulcherrima LS16 and two strains of the yeast-like fungus Aureobasidium pullulans LS30 and AU34-2-against infection by A. carbonarius and ochratoxin A (OTA) accumulation in wine grape berries was investigated in lab-scale experiments. The presence of wounds on grape skin dramatically favored infection of berries by A. carbonarius strain A1102, since unwounded berries showed very low levels of infection at all conditions of RH and temperature tested. Artificially wounded berries pre-treated with the biocontrol agents were inoculated with the ochratoxigenic A. carbonarius strain A1102 and were incubated for 5days at two levels of RH (60% and 100%) and three different temperatures (20, 25 and 30°C). The three biocontrol agents were able to prevent infections at 60% RH and 20°C. At 60% RH and 25°C only strain AU34-2 achieved some protection on day 5, whereas at 30°C a limited biocontrol efficacy was evident only up to day 2. At 100% RH, LS16, LS30 and AU34-2 showed effective protection of grape berries at 20°C until the 5th day of incubation. The three biocontrol agents achieved significant protection at higher temperatures only until the 2nd day after the beginning of the experiment: all three strains at 25°C, and only strain LS16 at 30°C. After 5days, the three biocontrol agents were able to significantly reduce the level of OTA in berries at all the conditions tested. This occurred even when protection from infection was not significant, except at 30°C and 100% of RH for all the three strains, and at 25°C and 100% of RH for strain LS16. The biocontrol agents displayed a higher rate of colonization on grape berries at 20 and 25°C than at 30°C. The higher value of RH (100%) appeared to increase the rate of colonization, in particular at 20 and 25°C. Taken together, our results emphasize the significant influence of environmental factors on the effectiveness of biocontrol against A. carbonarius as well as on OTA contamination in wine grape berries, and the need for biocontrol agents that can cope with the environmental conditions that are conducive to attack by A. carbonarius.     

Interactions of Spike-RBD of SARS-CoV-2 and Platelet Factor 4: New Insights in the Etiopathogenesis of Thrombosis

The rare but dangerous adverse events evidenced after massive vaccination against SARS-CoV-2 are represented by thrombosis and thrombocytopenia. The patients diagnosed with severe COVID-19 may develop a pro-thrombotic state with a much higher frequency, thus we decided to investigate the role of Spike protein (the only common product of the two conditions) or the anti-Spike antibodies in the etiopathogenesis of thrombosis. A pathogenic Platelet Factor 4 (PF4)-dependent syndrome, unrelated to the use of heparin therapy, has been reported after the administration of vaccines in the patients manifesting acute thrombocytopenia and thrombosis. Thus, we aimed at shedding light on the structural similarities of Spike of SARS-CoV-2 and PF4 on their eventual biochemical interactions and on the role of their specific antibodies. The similarities between PF4 and Spike-RBD proteins were evaluated by a comparison of the structures and by testing the cross-reactivity of their specific antibodies by ELISA assays. We found that the anti-Spike antibodies do not recognize PF4, on the contrary, the anti-PF4 antibodies show some cross-reactivity for Spike-RBD. More interestingly, we report for the first time that the PF4 and Spike-RBD proteins can bind each other. These data suggest that the interaction of the two proteins could be involved in the generation of anti-PF4 antibodies, their binding to Spike-RBD, which could lead to platelets aggregation due also to their high expression of ACE2.     

The Finely Coordinated Action of SSB and NurA/HerA Complex Strictly Regulates the DNA End Resection Process in Saccharolobus solfataricus

Generation of the 3′ overhang is a critical step during homologous recombination (HR) and replication fork rescue processes. This event is usually performed by a series of DNA nucleases and/or helicases. The nuclease NurA and the ATPase HerA, together with the highly conserved MRE11/RAD50 proteins, play an important role in generating 3′ single-stranded DNA during archaeal HR. Little is known, however, about HerA-NurA function and activation of this fundamental and complicated DNA repair process. Herein, we analyze the functional relationship among NurA, HerA and the single-strand binding protein SSB from Saccharolubus solfataricus. We demonstrate that SSB clearly inhibits NurA endonuclease activity and its exonuclease activities also when in combination with HerA. Moreover, we show that SSB binding to DNA is greatly stimulated by the presence of either NurA or NurA/HerA. In addition, if on the one hand NurA binding is not influenced, on the other hand, HerA binding is reduced when SSB is present in the reaction. In accordance with what has been observed, we have shown that HerA helicase activity is not stimulated by SSB. These data suggest that, in archaea, the DNA end resection process is governed by the strictly combined action of NurA, HerA and SSB.