CNBP Binds and Unfolds In Vitro G-Quadruplexes Formed in the SARS-CoV-2 Positive and Negative Genome Strands

The Coronavirus Disease 2019 (COVID-19) pandemic has become a global health emergency with no effective medical treatment and with incipient vaccines. It is caused by a new positive-sense RNA virus called severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2). G-quadruplexes (G4s) are nucleic acid secondary structures involved in the control of a variety of biological processes including viral replication. Using several G4 prediction tools, we identified highly putative G4 sequences (PQSs) within the positive-sense (+gRNA) and negative-sense (−gRNA) RNA strands of SARS-CoV-2 conserved in related betacoronaviruses. By using multiple biophysical techniques, we confirmed the formation of two G4s in the +gRNA and provide the first evidence of G4 formation by two PQSs in the −gRNA of SARS-CoV-2. Finally, biophysical and molecular approaches were used to demonstrate for the first time that CNBP, the main human cellular protein bound to SARS-CoV-2 RNA genome, binds and promotes the unfolding of G4s formed by both strands of SARS-CoV-2 RNA genome. Our results suggest that G4s found in SARS-CoV-2 RNA genome and its negative-sense replicative intermediates, as well as the cellular proteins that interact with them, are relevant factors for viral genes expression and replication cycle, and may constitute interesting targets for antiviral drugs development.     

Compositional characteristics,texture, shelf-life and sensory quality of snack crackers produced from non-traditional ingredients

The study was conducted to evaluate the effect of non-traditional and highly available and nutrient-dense ingredients in the production of value-added crackers. The crackers were prepared by combining partially defatted chia flour, wheat germ, quinoa, and oat. The antioxidant activity and the polyphenol content were 7.0–8.4 times higher in crackers produced with non-traditional ingredients compared to the control snack. The hydroperoxide value indicated a low oxidative deterioration of the oil. The sample with 10% of partially defatted chia flour was selected for shelf-life test under storage conditions and sensory evaluation. Modified atmosphere exerted a protective effect on the lipid stability regardless of the incorporation of BHT to the formulation. All the evaluated attributes scored highly during consumer acceptance. The formulated cracker presented a relevant content of protein, dietary fibre and omega-3 and -6 fatty acids. Based on these results, the crackers containing non-traditional ingredients resulted in a product with a good potential for both consumer acceptance and outstanding nutritional benefits.     

Intelligent measurement in clinical medicine — Analysis and examples

The paper analyses the concepts of intelligent measurement and instrumentation in relation to artificial intelligence. It describes examples of the application of intelligent measurement in clinical medicine: in particular, inferential measurement, the learning of patterns and measurement interpretation and its use in decision support in the total medical information system.     

Assessing the capability of broadband indices derived from Landsat 8 Operational Land Imager to monitor above ground biomass and salinity in semiarid saline environments of the Bahía Blanca Estuary,Argentina

In arid and semi-arid ecosystems, salinisation and desertification are the most common processes of land degradation, and satellite data may provide a valuable tool to assess land surface condition and vegetation status. The aim of this study was to evaluate the capability of Landsat 8 OLI (Operational Land Imager) remote sensing information and broadband indices derived from it, to monitor above ground biomass (AGB) and salinity in two different semiarid saline environments (unit a and unit b) in the Bahía Blanca Estuary. Unit a (Ua) is composed of bushes of Cyclolepis genistoides in association with Atriplex undulata and 41% of bare soil. Unit b (Ub) is composed of dense thickets of Allenrolfea patagonica in association with C. genistoides and 34% of bare soil. Pearson’s correlation analyses were performed between field estimates of AGB and salinity (soil salinity and interstitial water salinity) and remote sensing estimates. Satellite data include surface reflectance of individual bands, vegetation indices (NDVI [normalised difference vegetation index], SAVI [soil-adjusted vegetation index], MSAVI2 [modified soil-adjusted vegetation index], NDII [normalised difference infrared index], GNDVI [green normalised difference vegetation index], GRNDI [green-red normalised difference index], OSAVI [optimised soil-adjusted vegetation index], SR [simple ratio]), and salinity indices (SI1, SI2, SI3 [salinity index 1, 2 and 3, respectively] and BI [brightness index]). Correlation analyses involving AGB were performed twice; first considering all months and then again excluding the months with higher soil salinities. In Ua, soil adjusted vegetation indices SAVI and MSAVI2 showed to be suitable to detect changes in the total green AGB and C. genistoides green AGB (the major contributor to total green AGB). After excluding data from December and January (the months with the highest soil salinity), green AGB of A. undulata also showed a significant positive correlation with soil adjusted indices SAVI, MSAVI2 and OSAVI. Although proportionally this species was not a large contributor to the total biomass, it is characterised by a high leaf reflectance, which makes it suitable for biomass retrieval. In Ub, significant positive correlations were obtained between NDVI, SAVI, NDII, OSAVI and SR indices and the AGB green ratio, but significant negative correlations were obtained between A. patagonica red AGB and these vegetation indices. When December and January were excluded from the analysis the negative correlations between vegetation indices NDVI, OSAVI and SR and red AGB remained significant (r = ?0.68, ?0.76 and ?0.7, respectively). The positive correlations between these indices and AGB green ratio (r = 0.73, 0.78 and 0.75, respectively) remained significant as well. Significant negative correlations were also found between NDVI, NDII, GNDVI, OSAVI and SR indices and field salinity estimates. As soil salinisation induces A. patagonica reddening, red AGB and soil salinity covariate in the field, and the negative correlation with vegetation indices may be useful to retrieve information on both variables combined, which are indicative of water stress. Correlation analysis between field estimates of salinity and spectral salinity indices showed significant positive correlation for all the tested indices. The obtained results highlight the importance of a thoughtful selection of remote sensing indices to account for changes in vegetation biomass, especially in arid and semiarid environments particularly sensitive to desertification and salinisation. Also, ground truth cannot be overlooked, and field work is necessary to test index performance in every case.     

Particle size distribution of polypropylene produced in continuous polymerisation

A theoretical relationship has been developed between the particle size of polyolefin powders and that of the Ziegler-Natta catalyst from which they are derived. This has been extended to examine how the yield on catalyst and the residence time distribution in a continuous stirred tank reactor affect the particle size distribution. Experimental confirmation of these relationships has been obtained using TiCl₃ catalyst of near-spherical morphology prepared by low-temperature reduction of titanium tetrachloride with triethyl aluminium in solution.     

An optical NMR spectrometer for Larmor-beat detection and high-resolution POWER NMR

Optical nuclear magnetic resonance (ONMR) is a powerful probe of electronic properties in III-V semiconductors. Larmor-beat detection (LBD) is a sensitivity optimized, time-domain NMR version of optical detection based on the Hanle effect. Combining LBD ONMR with the line-narrowing method of POWER (perturbations observed with enhanced resolution) NMR further enables atomically detailed views of local electronic features in III-Vs. POWER NMR spectra display the distribution of resonance shifts or line splittings introduced by a perturbation, such as optical excitation or application of an electric field, that is synchronized with a NMR multiple-pulse time-suspension sequence. Meanwhile, ONMR provides the requisite sensitivity and spatial selectivity to isolate local signals within macroscopic samples. Optical NMR, LBD, and the POWER method each introduce unique demands on instrumentation. Here, we detail the design and implementation of our system, including cryogenic, optical, and radio-frequency components. The result is a flexible, low-cost system with important applications in semiconductor electronics and spin physics. We also demonstrate the performance of our systems with high-resolution ONMR spectra of an epitaxial AlGaAs/GaAs heterojunction. NMR linewidths down to 4.1 Hz full width at half maximum were obtained, a 10(3)-fold resolution enhancement relative any previous optically detected NMR experiment.     

Metabolic footprinting as a tool for discriminating between brewing yeasts

The characterization of industrial yeast strains by examining their metabolic footprints (exometabolomes) was investigated and compared to genome-based discriminatory methods. A group of nine industrial brewing yeasts was studied by comparing their metabolic footprints, genetic fingerprints and comparative genomic hybridization profiles. Metabolic footprinting was carried out by both direct injection mass spectrometry (DIMS) and gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), with data analysed by principal components analysis (PCA) and canonical variates analysis (CVA). The genomic profiles of the nine yeasts were compared by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, genetic fingerprinting using amplified fragment length polymorphism (AFLP) analysis and microarray comparative genome hybridizations (CGH). Metabolomic and genomic analysis comparison of the nine brewing yeasts identified metabolomics as a powerful tool in separating genotypically and phenotypically similar strains. For some strains discrimination not achieved genomically was observed metabolomically.     

Abrupt weaning reduces postweaning growth and is associated with alterations in gastrointestinal markers of development in dairy calves fed an elevated plane of nutrition during the preweaning period

The benefits of feeding elevated quantities of milk to dairy calves have been well established. However, there is a reluctance to adopt this method of feeding in commercial dairy production because of concerns around growth, health, and ruminal development during weaning. The objective of this study was to characterize the effect of an abrupt (0 d step-down) or gradual (12 d step-down) feeding scheme when calves are fed an elevated plane of nutrition (offered 1.35 kg of milk replacer/d). For this experiment, a total of 54 calves were randomly assigned to an abrupt or a gradual weaning protocol before weaning at 48 d of life. Calves were housed and sampled in individual pens for the duration of the experiment, and milk, starter, and straw intake were measured on a daily basis. Body weight was measured every 6 d, whereas blood, rumen fluid, and fecal samples were collected on d 36 (pre-step-down), 48 (preweaning), and 54 (postweaning) of the experiment. Although the growth rates of the step-down calves were lower from d 37 to weaning (0.62 ± 0.04 vs. 1.01 ± 0.04 kg/d), the postweaning average daily gain was greater compared with the group that was abruptly weaned (0.83 ± 0.06 vs. 0.22 ± 0.06 kg/d). Total ruminal volatile fatty acid was greater in the step-down group on the day of weaning (d 48; 59.80 ± 2.25 vs. 45.01 ± 2.25 mmol), whereas the fecal starch percentage was lower during postweaning compared with the abruptly weaned calves (d 54; 3.31 ± 0.76 vs. 6.34 ± 0.76%). Analysis of the digestive tract of bull calves on d 55 revealed minimal differences between gross anatomy measurements of gut compartments as well as no morphological differences in rumen papillae development, yet the total mass of rumen when full of contents was larger in the step-down calves (7.83 ± 0.78 vs. 6.02 ± 0.78 kg). Under the conditions of this study, the results showcase the benefits of a step-down feeding strategy from an overall energy balance standpoint, due to increased adaptation of the gastrointestinal tract preweaning.     

Novel Acyl Phosphate Mimics that Target PlsY,an Essential Acyltransferase in Gram‐Positive Bacteria

PlsY is a recently discovered acyltransferase that executes an essential step in membrane phospholipid biosynthesis in Gram‐ positive bacteria. By using a bioisosteric replacement approach to generate substrate‐based inhibitors of PlsY as potential novel antibacterial agents, a series of stabilized acyl phosphate mimetics, including acyl phosphonates, acyl α,α‐difluoromethyl phosphonates, acyl phosphoramides, reverse amide phosphonates, acyl sulfamates, and acyl sulfamides were designed and synthesized. Several acyl phosphonates, phosphoramides, and sulfamates were identified as inhibitors of PlsY from Streptococcus pneumoniae and Bacillus anthracis. As anticipated, these inhibitors were competitive inhibitors with respect to the acyl phosphate substrate. Antimicrobial testing showed the inhibitors to have generally weak activity against Gram‐positive bacteria with the exception of some acyl phosphonates, reverse amide phosphonates, and acyl sulfamates, which had potent activity against multiple strains of B. anthracis.