Probing the conformational dynamics of an Ago-RNA complex in water/methanol solution.

Argonaute (Ago) proteins mediate target recognition guiding miRNA to bind complementary mRNA primarily in the seed region. However, additional pairing can occur beyond the seed, forming a supplementary duplex that can contribute to the guide-target affinity. In order to shed light on the connection, between protein-RNA interactions and miRNA-mRNA seed and supplementary duplex mobility, we carried out molecular dynamics simulations at the microsecond time-scale using a different approach compared to the ones normally used. Until now, theoretical investigations with classical MD on Ago-RNA complexes have been focused primarily on pure water solvent, which mimics the natural environment of biological molecules. Here, we explored the conformational space of a human Ago2 (hAgo2) bound to the seed + supplementary miRNA-mRNA duplex, using the solvent environment as a molecular probe. MD simulations have been performed in a mixture of water/MeOH at a molar ratio of 70 : 30 as well as in pure water for comparison. Our findings revealed that the mixed solvent promotes protein RNA association, principally enhancing salt-linkages between basic amino acid side-chains and acidic phosphates of the sugar-phosphate backbone. The primary effect registered was the restriction of supplementary duplex flexibility and the stabilization of the miRNA 3' terminus. Interestingly, we observed that the influence of the solvent appears to have almost no impact on the conformation of the seed duplex.

Economic and social consequences of human mobility restrictions under COVID-19.

In response to the coronavirus disease 2019 (COVID-19) pandemic, several national governments have applied lockdown restrictions to reduce the infection rate. Here we perform a massive analysis on near-real-time Italian mobility data provided by Facebook to investigate how lockdown strategies affect economic conditions of individuals and local governments. We model the change in mobility as an exogenous shock similar to a natural disaster. We identify two ways through which mobility restrictions affect Italian citizens. First, we find that the impact of lockdown is stronger in municipalities with higher fiscal capacity. Second, we find evidence of a segregation effect, since mobility contraction is stronger in municipalities in which inequality is higher and for those where individuals have lower income per capita. Our results highlight both the social costs of lockdown and a challenge of unprecedented intensity: On the one hand, the crisis is inducing a sharp reduction of fiscal revenues for both national and local governments; on the other hand, a significant fiscal effort is needed to sustain the most fragile individuals and to mitigate the increase in poverty and inequality induced by the lockdown.

Revenue vs expenditure based fiscal consolidation: the pass-through from federal cuts to local taxes.

A growing literature emphasizes that the output effect of fiscal consolidation hinges on its composition, as the choice of increasing revenues vs cutting expenditure is not neutral. Existing studies, however, underscore the role of local governments in a federal setting. Indeed, transfer cuts at the central level might translate into higher local taxes, changing the effective composition of the fiscal adjustment. We evaluate this transmission mechanism in Italy, where municipalities below the threshold of 5,000 inhabitants were exempted from (large) transfer cuts in 2012. This allows us to implement a difference-in-discontinuities design in order to estimate the causal impact of transfer cuts on the composition of fiscal adjustment, also because tight fiscal rules impose a balanced budget on Italian municipalities. We find a pass-through mechanism by which local governments react to the contraction of intergovernmental grants by mainly increasing taxes rather than reducing spending. From a political economy perspective, this revenue based fiscal consolidation is driven by local governments with low electoral competition and low party fragmentation.

Integrated Geophysics and Geomatics Surveys in the Valley of the Kings.

Recent results within the framework of the collaborative project The Complete Geophysical Survey of the Valley of the Kings (VOK) (Luxor, Egypt) are reported in this article. In October 2018, a team of geomatics and geophysics researchers coordinated by the Polytechnic University of Turin worked side by side in the VOK. Topographic measurements in support of geophysical surveys and the achievement of a very large-scale 3D map of the Eastern VOK were the two main objectives of the geomatics campaign. Innovative 3D metric technologies and methods, based on terrestrial laser scanning (both static and mobile) and close-range photogrammetry were employed by the Geomatics team. The geophysical campaign focused on the acquisition of Electrical Resistivity Tomography (ERT), Ground Penetrating Radar (GPR) and high spatial density Geomagnetic (GM) data. ERT new data around KV62, both inverted in 2D sections and added to the previous ones to perform a new global 3D inversion, confirm the previous results showing both conductive and resistive anomalies that have to be explained. GPR timeslices showed some interesting features in the area in front of the KV2 entrance where GM gradient map also presents localized anomalies. In the area SSW of the KV2 the GM gradient maps evidenced also a large semicircular anomaly which, up to now, has no explanation. The potentialities of using magnetic techniques as a complement to other non-invasive techniques in the search for structures of archeological significance have been explored. The application of modern and innovative methods of 3D metric survey enabled to achieve a complete 3D mapping of what is currently visible in the valley. The integration of 2D/3D mapping data concerning visible elements and hypothetical anomalies, together with the recovering in the same global reference system of underground documentation pertaining to the Theban Mapping Project, prefigure the enhancement of multi-temporal site representation. This strategy enables the fruition development of the already discovered archaeological heritage, using modern criteria of valorization and conservation.

High-Intensity Focused Ultrasonography and Radiofrequency Ablation of Renal Cell Carcinoma Arisen in Transplanted Kidneys: Single-Center Experience With Long-Term Follow-Up and Review of Literature.

The purpose of this article is to retrospectively evaluate the long-term outcome of patients treated with percutaneous thermoablation for renal cell carcinomas that have arisen in kidney grafts. Between April 2008 and February 2011, we treated 3 patients with renal cell carcinoma on a transplanted kidney: 2 cases were treated with high-intensity focused ultrasonography and 1 patient with radio frequency ablation. Postprocedural ultrasonography did not reveal any complications, and contrast-enhanced ultrasonography showed an avascular area in the treated nodules. None of the patients had recurrent tumors during a long-term clinical and radiologic follow-up (81, 73, and 43 months, respectively).

Infectivity and transmission of Xylellua fastidiosa by Philaenus spumarius (Hemiptera: Aphrophoridae) in Apulia, Italy.

Discovery of Xylella fastidiosa from olive trees with "Olive quick decline syndrome" in October 2013 on the west coast of the Salento Peninsula prompted an immediate search for insect vectors of the bacterium. The dominant xylem-fluid feeding hemipteran collected in olive orchards during a 3-mo survey was the meadow spittlebug, Philaenus spumarius (L.) (Hemiptera: Aphrophoridae). Adult P. spumarius, collected in November 2013 from ground vegetation in X. fastidiosa-infected olive orchards, were 67% (40 out of 60) positive for X. fastidiosa by polymerase chain reaction (PCR) assays. Euscelis lineolatus Brullé were also collected but tested negative for the pathogen. Transmission tests with P. spumarius collected from the Salento area were, therefore, conducted. After a 96-h inoculation access period with 8 to 10 insects per plant and a 30-d incubation period, PCR results showed P. spumarius transmitted X. fastidiosa to two of five periwinkle plants but not to the seven olive plants. Sequences of PCR products from infected periwinkle were identical with those from X. fastidiosa-infected field trees. These data showed P. spumarius as a vector of X. fastidiosa strain infecting olives trees in the Salento Peninsula, Italy.

Correction: Lozano-Soria et al. Volatile Organic Compounds from Entomopathogenic and Nematophagous Fungi, Repel Banana Black Weevil (Cosmopolites sordidus). Insects 2020, 11, 509.

In the original publication [...].

Native Entomopathogenic Fungi Isolated from Rhynchophorus palmarum (Linnaeus, 1758) in Northeast Brazil.

Both palm weevils, the South American (Rhynchophorus palmarum) (SAPW) and the red palm weevil (R. ferrugineus, RPW), are present in South America, affecting commercial, ornamental, and native palms. These pests oviposit and thrive on selected Arecaceae. R. palmarum mainly infests coconut (Cocos nucifera), oil palms (Elaeis guineensis), and other ornamental and native palms in America, causing a significant social impact on growers. The weevils fulfill a significant ectosymbiotic macro- and microorganism role in the first period of larval development, worsening the damage which, during this period, is not yet apparent. Palm protection in the Brazilian context suggests the use of indigenous agents for microbiological biocontrol. This research identifies three Brazilian Beauveria bassiana isolates: CVAD01, CVAD02, and CVAD06. The results suggest that the strain's impact on R. palmarum can also be compared with that of the commercial strain Beauveria bassiana. Phylogenetic analysis allowed the delimitation of species of Beauveria (Hypocreales). Pathogenicity tests caused significant mortality in R. palmarum. The isolates CVAD01, CVAD02, and CVADO6 showed high pathogenicity between 7 and 21 days, with mortality rates between 90 and 100%, suggesting that they may be effective biological control agents of R. palmarum in the field when used, within available means, to mitigate the impact of R. palmarum and R. ferrugineus in South America.

Bioinformatic survey of CRISPR loci across 15 Serratia species.

The Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins (CRISPR-Cas) system of prokaryotes is an adaptative immune defense mechanism to protect themselves from invading genetic elements (e.g., phages and plasmids). Studies that describe the genetic organization of these prokaryotic systems have mainly reported on the Enterobacteriaceae family (now reorganized within the order of Enterobacterales). For some genera, data on CRISPR-Cas systems remain poor, as in the case of Serratia (now part of the Yersiniaceae family) where data are limited to a few genomes of the species marcescens. This study describes the detection, in silico, of CRISPR loci in 146 Serratia complete genomes and 336 high-quality assemblies available for the species ficaria, fonticola, grimesii, inhibens, liquefaciens, marcescens, nematodiphila, odorifera, oryzae, plymuthica, proteomaculans, quinivorans, rubidaea, symbiotica, and ureilytica. Apart from subtypes I-E and I-F1 which had previously been identified in marcescens, we report that of I-C and the I-E unique locus 1, I-E*, and I-F1 unique locus 1. Analysis of the genomic contexts for CRISPR loci revealed mdtN-phnP as the region mostly shared (grimesii, inhibens, marcescens, nematodiphila, plymuthica, rubidaea, and Serratia sp.). Three new contexts detected in genomes of rubidaea and fonticola (puu genes-mnmA) and rubidaea (osmE-soxG and ampC-yebZ) were also found. The plasmid and/or phage origin of spacers was also established.

Brindley's Glands Volatilome of the Predator Zelus renardii Interacting with Xylella Vectors.

Alien species must adapt to new biogeographical regions to acclimatise and survive. We consider a species to have become invasive if it establishes negative interactions after acclimatisation. Xylella fastidiosa Wells, Raju et al., 1986 (XF) represents Italy's and Europe's most recent biological invasion. In Apulia (southern Italy), the XF-encountered Philaenus spumarius L. 1758 (Spittlebugs, Hemiptera: Auchenorrhyncha) can acquire and transmit the bacterium to Olea europaea L., 1753. The management of XF invasion involves various transmission control means, including inundative biological control using Zelus renardii (ZR) Kolenati, 1856 (Hemiptera: Reduviidae). ZR is an alien stenophagous predator of Xylella vectors, recently entered from the Nearctic and acclimated in Europe. Zelus spp. can secrete semiochemicals during interactions with conspecifics and prey, including volatile organic compounds (VOCs) that elicit conspecific defence behavioural responses. Our study describes ZR Brindley's glands, present in males and females of ZR, which can produce semiochemicals, eliciting conspecific behavioural responses. We scrutinised ZR secretion alone or interacting with P. spumarius. The ZR volatilome includes 2-methyl-propanoic acid, 2-methyl-butanoic acid, and 3-methyl-1-butanol, which are consistent for Z. renardii alone. Olfactometric tests show that these three VOCs, individually tested, generate an avoidance (alarm) response in Z. renardii. 3-Methyl-1-butanol elicited the highest significant repellence, followed by 2-methyl-butanoic and 2-methyl-propanoic acids. The concentrations of the VOCs of ZR decrease during the interaction with P. spumarius. We discuss the potential effects of VOC secretions on the interaction of Z. renardii with P. spumarius.

Considerations on chemical composition of psammoma bodies: Automated detection strategy by infrared microspectroscopy in ovarian and thyroid cancer tissues.

Ectopic calcifications are observed in many soft tissues and are associated with several diseases, including cancer. The mechanism of their formation and the correlation with disease progression are often unclear. Detailed knowledge of the chemical composition of these inorganic formations can be very helpful in better understanding their relationship with unhealthy tissue. In addition, information on microcalcifications can be very useful for early diagnosis and provide insight into prognosis. In this work the chemical composition of psammoma bodies (PBs) found in tissues of human ovarian serous tumors was examined. The analysis using Micro Fourier Transform Infrared Spectroscopy (micro-FTIR) revealed that these microcalcifications contain amorphous calcium carbonate phosphate. Moreover, some PB grains showed the presence of phospholipids. This interesting result corroborates the proposed formation mechanism reported in many studies according to which ovarian cancer cells switch to a calcifying phenotype by inducing the deposition of calcifications. In addition, other techniques as X-ray Fluorescence Spectroscopy (XRF), Inductively Coupled Plasma Optical Emission Spectroscopy(ICP-OES) and Scanning electron microscopy (SEM) with Energy Dispersive X-ray Spectroscopy (EDX) were performed on the PBs from ovary tissues to determine the elements present. The PBs found in ovarian serous cancer showed a composition comparable to PBs isolated from papillary thyroid. Based on the chemical similarity of IR spectra, using micro-FTIR spectroscopy combined with multivariate analysis, an automatic recognition method was constructed. With this prediction model it was possible to identify PBs microcalcifications in tissues of both ovarian cancers, regardless of tumor grade, and thyroid cancer with high sensitivity. Such approach could become a valuable tool for routine macrocalcification detection because it eliminates sample staining, and the subjectivity of conventional histopathological analysis.

"Ectomosphere": Insects and Microorganism Interactions.

This study focuses on interacting with insects and their ectosymbiont (lato sensu) microorganisms for environmentally safe plant production and protection. Some cases help compare ectosymbiont microorganisms that are insect-borne, -driven, or -spread relevant to endosymbionts' behaviour. Ectosymbiotic bacteria can interact with insects by allowing them to improve the value of their pabula. In addition, some bacteria are essential for creating ecological niches that can host the development of pests. Insect-borne plant pathogens include bacteria, viruses, and fungi. These pathogens interact with their vectors to enhance reciprocal fitness. Knowing vector-phoront interaction could considerably increase chances for outbreak management, notably when sustained by quarantine vector ectosymbiont pathogens, such as the actual Xylella fastidiosa Mediterranean invasion episode. Insect pathogenic viruses have a close evolutionary relationship with their hosts, also being highly specific and obligate parasites. Sixteen virus families have been reported to infect insects and may be involved in the biological control of specific pests, including some economic weevils. Insects and fungi are among the most widespread organisms in nature and interact with each other, establishing symbiotic relationships ranging from mutualism to antagonism. The associations can influence the extent to which interacting organisms can exert their effects on plants and the proper management practices. Sustainable pest management also relies on entomopathogenic fungi; research on these species starts from their isolation from insect carcasses, followed by identification using conventional light or electron microscopy techniques. Thanks to the development of omics sciences, it is possible to identify entomopathogenic fungi with evolutionary histories that are less-shared with the target insect and can be proposed as pest antagonists. Many interesting omics can help detect the presence of entomopathogens in different natural matrices, such as soil or plants. The same techniques will help localize ectosymbionts, localization of recesses, or specialized morphological adaptation, greatly supporting the robust interpretation of the symbiont role. The manipulation and modulation of ectosymbionts could be a more promising way to counteract pests and borne pathogens, mitigating the impact of formulates and reducing food insecurity due to the lesser impact of direct damage and diseases. The promise has a preventive intent for more manageable and broader implications for pests, comparing what we can obtain using simpler, less-specific techniques and a less comprehensive approach to Integrated Pest Management (IPM).

Effect of Precursors on the Electrochemical Properties of Mixed RuOx/MnOx Electrodes Prepared by Thermal Decomposition.

Growing thin layers of mixed-metal oxides on titanium supports allows for the preparation of versatile electrodes that can be used in many applications. In this work, electrodes coated with thin films of ruthenium (RuOx) and manganese oxide (MnOx) were fabricated via thermal decomposition of a precursor solution deposited on a titanium substrate by spin coating. In particular, we combined different Ru and Mn precursors, either organic or inorganic, and investigated their influence on the morphology and electrochemical properties of the materials. The tested salts were: Ruthenium(III) acetylacetonate (Ru(acac)3), Ruthenium(III) chloride (RuCl3·xH2O), Manganese(II) nitrate (Mn(NO3)2·4H2O), and Manganese(III) acetylacetonate (Mn(acac)3). After fabrication, the films were subjected to different characterization techniques, including scanning electron microscopy (SEM), polarization analysis, open-circuit potential (OCP) measurements, electrochemical impedance spectroscopy (EIS), linear sweep voltammetry (LSV), cyclic voltammetry (CV), and galvanostatic charge-discharge (GCD) experiments. The results indicate that compared to the others, the combination of RuCl3 and Mn(acac) produces fewer compact films, which are more susceptible to corrosion, but have outstanding capacitive properties. In particular, this sample exhibits a capacitance of 8.3 mF cm-2 and a coulombic efficiency of higher than 90% in the entire range of investigated current densities.

Zelus renardii Roaming in Southern Italy.

This study collects data from the literature and updates our Zelus renardii Kolenati, 1856 (Leafhopper Assassin Bug, LAB) prey knowledge. The literature consists of ca. 170 entries encompassing the years 1856 to 2021. This reduviid originated in the Nearctic region, but has entered and acclimatised in many Mediterranean countries. Our quantitative predation experiments-in the laboratory on caged plants plus field or environmental observations-confirm that LAB prefers a selected array of prey. Laboratory predation tests on living targets (Hemiptera, Coleoptera, Diptera, and Hymenoptera) agree with the literature. Zelus renardii prefers comparatively large, highly mobile, and readily available prey. LAB preferences on available hemipterans targets suggest that Zelus renardii is a good inundative biocontrol agent for Xylella fastidiosapauca ST53 infections. LAB also prey on other important olive pests, such as Bactrocera oleae. Therefore, Zelus renardii is a major integrated pest management (IPM) component to limit Xylella fastidiosa pandemics and other pest invasions.

Protein Sources Alternative to Meat: State of the Art and Involvement of Fermentation.

Meat represents an important protein source, even in developing countries, but its production is scarcely sustainable, and its excessive consumption poses health issues. An increasing number of Western consumers would replace, at least partially, meat with alternative protein sources. This review aims at: (i) depicting nutritional, functional, sensory traits, and critical issues of single-cell proteins (SCP), filamentous fungi, microalgae, vegetables (alone or mixed with milk), and insects and (ii) displaying how fermentation could improve their quality, to facilitate their use as food items/ingredients/supplements. Production of SCP (yeasts, filamentous fungi, microalgae) does not need arable land and potable water and can run continuously, also using wastes and byproducts. Some filamentous fungi are also consumed as edible mushrooms, and others are involved in the fermentation of traditional vegetable-based foods. Cereals, pseudocereals, and legumes may be combined to offer an almost complete amino acid profile. Fermentation of such vegetables, even in combination with milk-based products (e.g., tarhana), could increase nutrient concentrations, including essential amino acids, and improve sensory traits. Different insects could be used, as such or, to increase their acceptability, as ingredient of foods (e.g., pasta). However, insects as a protein source face with safety concerns, cultural constraints, and a lack of international regulatory framework.

Identification of remodeled collagen fibers in tumor stroma by FTIR Micro-spectroscopy: A new approach to recognize the colon carcinoma.

The tumor stroma plays a pivotal role in colon cancer genesis and progression. It was observed that collagen fibers in the extracellular matrix (ECM) of cancer stroma, undergo a strong remodeling. These fibrous proteins result more aligned and compact than in physiological conditions, creating a microenvironment that favors cancer development. In this work, micro-FTIR spectroscopy was applied to investigate the chemical modifications in the tumor stroma. Using Fuzzy C-means clustering, mean spectra from diseased and normal stroma were compared and collagen was found to be responsible for the main differences between them. Specifically, the modified absorptions at 1203, 1238, 1284 cm-1 and 1338 cm-1 wavenumbers, were related to the amide III band and CH2 bending of side chains. These signals are sensitive to the interactions between the α-chains in the triple helices of collagen structure. This provided robust chemical evidence that in cancer ECM, collagen fibers are more parallelized, stiff and ordered than in normal tissue. Principal Component Analysis (PCA) applied to the spectra from malignant and normal stroma confirmed these findings. Using LDA (Linear Discriminant Analysis) classification, the absorptions 1203, 1238, 1284 and 1338 cm-1 were examined as spectral biomarkers, obtaining quite promising results. The use of a PCA-LDA prediction model on samples with moderate tumor degree further showed that the stroma chemical modifications are more indicative of malignancy compared to the epithelium. These preliminary findings have shown that micro-FTIR spectroscopy, focused on collagen signals, could become a promising tool for colon cancer diagnosis.

Quantitative Real-Time PCR Based on SYBR Green Technology for the Identification of Philaenus italosignus Drosopoulos & Remane (Hemiptera Aphrophoridae).

The use of molecular tools to identify insect pests is a critical issue, especially when rapid and reliable tests are required. We proposed a protocol based on qPCR with SYBR Green technology to identify Philaenus italosignus (Hemiptera, Aphrophoridae). The species is one of the three spittlebugs able to transmit Xylella fastidiosa subsp. pauca ST53 in Italy, together with Philaenus spumarius and Neophilaenus campestris. Although less common than the other two species, its identification is key to verifying which role it can play when locally abundant. The proposed assay shows analytical specificity being inclusive with different populations of the target species and exclusive with non-target taxa, either taxonomically related or not. Moreover, it shows analytical sensibility, repeatability, and reproducibility, resulting in an excellent candidate for an official diagnostic method. The molecular test can discriminate P. italosignus from all non-target species, including the congeneric P. spumarius.

Aphrophoridae Role in Xylella fastidiosa subsp. pauca ST53 Invasion in Southern Italy.

The Philaenus spumarius L. (Hemiptera Aphrophoridae) is a xylem-sap feeder vector that acquires Xylella fastidiosa subsp. pauca ST53 during feeding on infected plants. The bacterium is the plant pathogen responsible for olive quick decline syndrome that has decimated olive trees in Southern Italy. Damage originates mainly from the insect vector attitude that multiplies the pathogen potentialities propagating Xf in time and space. The principal action to manage insect-borne pathogens and to contain the disease spread consists in vector and transmission control. The analysis of an innovative and sustainable integrated pest management quantitative strategy that targets the vector and the infection by combining chemical and physical control means demonstrates that it is possible to stop the Xylella invasion. This review updates the available topics addressing vectors' identification, bionomics, infection management, and induced disease by Xylella invasion to discuss major available tools to mitigate the damage consequent to the disease.

A Simple Cerium Coating Strategy for Titanium Oxide Nano-tubes' Bioactivity Enhancement.

Despite the well-known favorable chemical and mechanical properties of titanium-based materials for orthopedic and dental applications, poor osseointegration of the implants, bacteria adhesion, and excessive inflammatory response from the host remain major problems to be solved. Here, the antioxidant and anti-inflammatory enzyme-like abilities of ceria (CeOx) were coupled to the advantageous features of titanium nanotubes (TiNTs). Cost-effective and fast methods, such as electrochemical anodization and drop casting, were used to build active surfaces with enhanced bioactivity. Surface composition, electrochemical response, and in vitro ability to induce hydroxyapatite (HA) precipitation were evaluated. The amount of cerium in the coating did not significantly affect wettability, yet a growing ability to induce early HA precipitation from simulated body fluid (SBF) was observed as the oxide content at the surface increased. The presence of 4%wt CeOx was also able to stimulate rapid HA maturation in a (poorly) crystalline form, indicating an interesting potential to induce rapid in vivo osseointegration process.