Biochemical and spectroscopic properties of Brucella microti glutamate decarboxylase, a key component of the glutamate-dependent acid resistance system

In orally acquired bacteria, the ability to counteract extreme acid stress (pH < 2.5) ensures survival during transit through the animal host stomach. In several neutralophilic bacteria, the glutamate-dependent acid resistance system (GDAR) is the most efficient molecular system in conferring protection from acid stress. In Escherichia coli its structural components are either of the two glutamate decarboxylase isoforms (GadA, GadB) and the antiporter, GadC, which imports glutamate and exports γ-aminobutyrate, the decarboxylation product. The system works by consuming protons intracellularly, as part of the decarboxylation reaction, and exporting positive charges via the antiporter. Herein, biochemical and spectroscopic properties of GadB from Brucella microti (BmGadB), a Brucella species which possesses GDAR, are described. B. microti belongs to a group of lately described and atypical brucellae that possess functional gadB and gadC genes, unlike the most well-known "classical" Brucella species, which include important human pathogens. BmGadB is hexameric at acidic pH. The pH-dependent spectroscopic properties and activity profile, combined with in silico sequence comparison with E. coli GadB (EcGadB), suggest that BmGadB has the necessary structural requirements for the binding of activating chloride ions at acidic pH and for the closure of its active site at neutral pH. On the contrary, cellular localization analysis, corroborated by sequence inspection, suggests that BmGadB does not undergo membrane recruitment at acidic pH, which was observed in EcGadB. The comparison of GadB from evolutionary distant microorganisms suggests that for this enzyme to be functional in GDAR some structural features must be preserved

Handheld-Impedance-Measurement System with seven-decade capability and potentiostatic function

This paper describes design and test of a new impedance-measurement system for nonlinear devices that exhibits a seven-decade range and works down to a frequency of 0.01 Hz. The system is specifically designed for electrochemical measurements, but the proposed architecture can be employed in many other fields where flexible signal generation and analysis are required. The system employs an unconventional signal generator based on two pulsewidth modulation (PWM) oscillators and an autocalibration system that allows uncertainties of less than 3% to be obtained over a range of 1 kΩ to 100 GΩ. A synchronous demodulation processing allows the noise superimposed to the low-amplitude input signals to be made negligibl

Electrochemical characterization of innovative hybrid coatings for metallic artefacts

In this paper, an electrochemical characterization of two different hybrid coatings is presented, with the specific aim of studying their corrosion protection behavior and better understanding their possible application in the cultural heritage field. The two formulations under study were epoxy resin containing silica nanoparticles and epoxy resin containing graphene oxide. Electrochemical Impedance Spectroscopy (EIS) and Scanning ElectroChemical Microscopy (SECM) were used to compare the electrochemical behavior of the two coatings and highlight their failure mechanism when immersed in an electrolytic solution containing chlorides. The investigation highlighted the good corrosion protective properties of both coatings; and moreover, thanks to the joined use of the two described analytical techniques, the different water uptake of the two solutions was studied, together with the different evolution of the coating surface morphology when immersed in the electrolytic solution

Impact of urbanization trends on production of key staple crops

Urbanization has appropriated millions of hectares of cropland, and this trend will persist as cities continue to expand. We estimate the impact of this conversion as the amount of land needed elsewhere to give the same yield potential as determined by differences in climate and soil properties. Robust spatial upscaling techniques, well-validated crop simulation models, and soil, climate, and cropping system databases are employed with a focus on populous countries with high rates of land conversion. We find that converted cropland is 30–40% more productive than new cropland, which means that projection of food production potential must account for expected cropland loss to urbanization. Policies that protect existing farmland from urbanization would help relieve pressure on expansion of agriculture into natural ecosystems

Decipher the glioblastoma microenvironment: The first milestone for new groundbreaking therapeutic strategies

Glioblastoma (GBM) is the most common primary malignant brain tumour in adults. Despite the combination of novel therapeutical approaches, it remains a deadly malignancy with an abysmal prognosis. GBM is a polymorphic tumour from both molecular and histological points of view. It consists of different malignant cells and various stromal cells, contributing to tumour initiation, progression, and treatment response. GBM’s microenvironment is multifaceted and is made up of soluble factors, extracellular matrix components, tissue-resident cell types (e.g., neurons, astrocytes, endothelial cells, pericytes, and fibroblasts) together with resident (e.g., microglia) or recruited (e.g., bone marrow-derived macrophages) immune cells. These latter constitute the so-called immune microenvironment, accounting for a substantial GBM’s tumour volume. Despite the abundance of immune cells, an intense state of tumour immunosuppression is promoted and developed; this represents the significant challenge for cancer cells’ immune-mediated destruction. Though literature data suggest that distinct GBM’s subtypes harbour differences in their microenvironment, its role in treatment response remains obscure. However, an in-depth investigation of GBM’s microenvironment may lead to novel therapeutic opportunities to improve patients’ outcomes. This review will elucidate the GBM’s microenvironment composition, highlighting the current state of the art in immunotherapy approaches. We will focus on novel strategies of active and passive immunotherapies, including vaccination, gene therapy, checkpoint blockade, and adoptive T-cell therapies

Characterization of the water diffusion in GEM foil material

Systematic studies on the GEM foil material are performed to measure the moisture diffusion rate and saturation level.These studies are important because the presence of this compound inside the detector’s foil can possibly change its mechanical and electrical properties,and in such a way,the detector performance can be affected.To understand this phenomenon,a model is developed with COMSOL Multiphysicsv.4.3 which described the adsorption and diffusion within the geometry of GEM foil,the concentration profiles and the time required to saturate the foil.The COMSOL model is verified by experimental observations on a GEM foil sample.This note will describe the model and its experimental verification results

Analysis of the DNA methylation pattern of the promoter region of calcitonin gene-related peptide 1 gene in patients with episodic migraine: An exploratory case-control study

Recent studies suggested that epigenetic mechanisms, including DNA methylation, may be involved in migraine pathogenesis. The calcitonin gene-related peptide (CGRP), encoded by calcitonin gene-related peptide 1 (CALCA) gene, plays a key role in the disease. The aim of the study was to evaluate DNA methylation of CALCA gene in patients with episodic migraine. 22 patients with episodic migraine (F/M 15/7, mean age 39.7 ± 13.4 years) and 20 controls (F/M 12/8, mean age 40.5 ± 14.8 years) were recruited. Genomic DNA was extracted from peripheral blood. Cytosine-to-thymine conversion was obtained with sodium bisulfite. The methylation pattern of two CpG islands in the promoter region of CALCA gene was analyzed. No difference of methylation of the 30 CpG sites at the distal region of CALCA promoter was observed between migraineurs and controls. Interestingly, in patients with episodic migraine the methylation level was lower in 2 CpG sites at the proximal promoter region (CpG −1461, p = 0.037, and −1415, p = 0.035, respectively). Furthermore, DNA methylation level at different CpG sites correlates with several clinical characteristics of the disease, as age at onset, presence of nausea/vomiting, depression and anxiety (p < 0.05). In conclusion, we found that DNA methylation profile in two CpG sites at the proximal promoter region of CALCA is lower in migraineurs when compared to controls. Intriguingly, the −1415 hypomethylated unit is located at the CREB binding site, a nuclear transcription factor. In addition, we found a correlation between the level of CALCA methylation and several clinical features of migraine. Further studies with larger sample size are needed to confirm these results

Spatial frameworks for robust estimation of yield gaps

Food security interventions and policies need reliable estimates of crop production and the scope to enhance production on existing cropland. Here we assess the performance of two widely used ‘top-down’ gridded frameworks (Global Agro-ecological Zones and Agricultural Model Intercomparison and Improvement Project) versus an alternative ‘bottom-up’ approach (Global Yield Gap Atlas). The Global Yield Gap Atlas estimates extra production potential locally for a number of sites representing major breadbaskets and then upscales the results to larger spatial scales. We find that estimates from top-down frameworks are alarmingly unlikely, with estimated potential production being lower than current farm production at some locations. The consequences of using these coarse estimates to predict food security are illustrated by an example for sub-Saharan Africa, where using different approaches would lead to different prognoses about future cereal self-sufficiency. Our study shows that foresight about food security and associated agriculture research priority setting based on yield potential and yield gaps derived from top-down approaches are subject to a high degree of uncertainty and would benefit from incorporating estimates from bottom-up approaches

PECVD Coatings for Magnesium Casting Protection

In this study SiO2-like films have been deposited by Plasma Enhanced Chemical Vapour Deposition(PECVD) on Mg alloys employed in the automotive industry in different experimental conditions in orderto optimize the protective effectiveness of the deposited layers. The experimental apparatus used was acapacitive coupled (CC) parallel-plate-reactor with an asymmetric electrode configuration, theelectromagnetic radiation was applied to a gas mixture containing tetraetoxysilane (TEOS), oxygen andargon in different ratios. The deposition was performed at 100 mTorr of pressure, by varying the inputpower, on as cast and polished samples submitted to a pre-treatment step in H2-containing plasma. A setof Mg alloy specimens has been treated with a standard anodising process (AP).From the overall experimental results, PECVD appears an attractive surface modification techniquewhich allows, through the deposition of SiO2-like films, to increase the corrosion resistance of Mg alloys,usually protected with anodising treatment or with chromium-based coatings. The check of thetransferability of deposition process to a medium scale range is now running. Magnesium framesrepresentative of real automotive parts coated in a PECVD industrial chamber, assembled and paintedare submitted to accelerated corrosion test in order to verify the corrosion resistance of these paintedelements manufactured following the production steps of a real component