Identification of Host Cellular Protein Substrates of SARS-COV-2 Main Protease

The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease-19 (COVID-19) being associated with severe pneumonia. Like with other viruses, the interaction of SARS-CoV-2 with host cell proteins is necessary for successful replication, and cleavage of cellular targets by the viral protease also may contribute to the pathogenesis, but knowledge about the human proteins that are processed by the main protease (3CLpro) of SARS-CoV-2 is still limited. We tested the prediction potentials of two different in silico methods for the identification of SARS-CoV-2 3CLpro cleavage sites in human proteins. Short stretches of homologous host-pathogen protein sequences (SSHHPS) that are present in SARS-CoV-2 polyprotein and human proteins were identified using BLAST analysis, and the NetCorona 1.0 webserver was used to successfully predict cleavage sites, although this method was primarily developed for SARS-CoV. Human C-terminal-binding protein 1 (CTBP1) was found to be cleaved in vitro by SARS-CoV-2 3CLpro, the existence of the cleavage site was proved experimentally by using a His₆-MBP-mEYFP recombinant substrate containing the predicted target sequence. Our results highlight both potentials and limitations of the tested algorithms. The identification of candidate host substrates of 3CLpro may help better develop an understanding of the molecular mechanisms behind the replication and pathogenesis of SARS-CoV-2.     

On the effects of the packet size distribution on the packet loss process

Real-time multimedia applications have to use forward error correction (FEC) anderror concealment techniques to cope with losses in today’s best-effort Internet. The efficiency of these solutions is known however to depend on the correlation between losses in the media stream. In this paper we investigate how the packet size distribution affects the packet loss process, that is, the distribution of the number of lost packets in a block, the related FEC performance and the average loss run length. We present mathematical models for the loss process of the MMPP+M/D/1/K and the MMPP+M/M/1/K queues; we validate the models via simulations, and compare the results to simulation results with an MPEG-4 coded video trace. We conclude that the deterministic packet size distribution (PSD) not only results in lower stationary loss probability than the exponential one, but also gives a less correlated loss process, both at a particular average link load and at a particular stationary loss probability as seen by the media stream.Our results show that for applications that can only measure the packet loss probability, the effects of the PSD on FEC performance are higher in access networks, where a single multimedia stream might affect the multiplexing behavior. Our results show that the effects of the PSD on FEC performance are higher in access networks, where a single multimedia stream might affect the multiplexing behavior and thus can improve the queuing performance by decreasing the variance of its PSD.     

Characterization of the Residence Time Distribution in Spray Dryers

In this work it is presented a study on the residence time distribution (RTD) of particles in a co-current pilot-plant spray dryer operated with a rotary atomization system. A nuclear technique is applied to investigate the RTD responses of spray dryers. The methodology is based on the injection of a radioisotope tracer in the feed stream followed by the monitoring of its concentration at the outlet stream. The experiments were performed during the drying of aqueous suspensions of gadolinium oxide. The RTD responses obtained experimentally presented good reproducibility, indicating that the technique applied is well suited to investigating fluid-dynamics of spray dryers. In addition to the experimental investigation, a mathematical model was used to describe the RTD experimental curves.     

TRPM7-Mediated Calcium Transport in HAT-7 Ameloblasts

TRPM7 plays an important role in cellular Ca²⁺, Zn²⁺ and Mg²⁺ homeostasis. TRPM7 channels are abundantly expressed in ameloblasts and, in the absence of TRPM7, dental enamel is hypomineralized. The potential role of TRPM7 channels in Ca²⁺ transport during amelogenesis was investigated in the HAT-7 rat ameloblast cell line. The cells showed strong TRPM7 mRNA and protein expression. Characteristic TRPM7 transmembrane currents were observed, which increased in the absence of intracellular Mg²⁺ ([Mg²⁺]_i), were reduced by elevated [Mg²⁺]_i, and were inhibited by the TRPM7 inhibitors NS8593 and FTY720. Mibefradil evoked similar currents, which were suppressed by elevated [Mg²⁺]_i, reducing extracellular pH stimulated transmembrane currents, which were inhibited by FTY720. Naltriben and mibefradil both evoked Ca²⁺ influx, which was further enhanced by the acidic intracellular conditions. The SOCE inhibitor BTP2 blocked Ca²⁺ entry induced by naltriben but not by mibefradil. Thus, in HAT-7 cells, TRPM7 may serves both as a potential modulator of Orai-dependent Ca²⁺ uptake and as an independent Ca²⁺ entry pathway sensitive to pH. Therefore, TRPM7 may contribute directly to transepithelial Ca²⁺ transport in amelogenesis.     

Design and analysis of single-factor experiments: Analysis of variance of the effect of rice husk ash and commercial fillers in NR compounds

Summary White rice husk ash (WRHA) and black rice husk ash (BRHA) were incorporated into natural rubber (NR) using a laboratory-sized two-roll mill. A conventional vulcanization system was used for curing and physical tests of the NR vulcanized involved determining of tensile and tear resistances. For comparison purposes, precipitated silica (Zeosil-175) and carbon black (N774) were used too. Using the analysis of variance of single-factor experiments, it can be concluded that: BRHA is non-reinforcing filler and its use is limited to 20 phr; WRHA is semi-reinforcing filler and the variation of filler loading (0 up to 50 phr) causes the maximum variation upon tensile strength of NR compounds; and, that although carbon black and silica are reinforcing fillers, a real reinforcement is reached up to 20 phr for tensile strength.     

Evaluation of the removal of n-butanol vapor by the poly(lactic acid)-zeolite-TiO2 composite and formation of by-products

The use of polymeric films incorporated with zeolite-TiO₂ composites associated with UV radiation can be an alternative in the removal of volatile organic compounds (VOCs) through the adsorption and photodegradation processes. This study produced poly(lactic acid) (PLA) films incorporated with 13× zeolite, TiO₂, and 13×-TiO₂ zeolite composite to remove n-butanol and evaluate the by-products generated in the process. The results showed that 13× zeolite and TiO₂ added individually or as a composite to PLA, gave the polymer matrix a significant increase in the removal capacity of n-butanol. The best performance was presented by the zeolite-TiO₂, composite, confirming a synergistic effect. However, the formation of CO and CO₂ exceeded the expected values, with the verification that the polymeric matrix underwent photodegradation action by TiO₂. The polymeric film only containing zeolite is the most suitable for the removal of VOCs, as it did not present degradation of the PLA, generating a lower concentration of by-products.     

pH sensitive phosphate crosslinked films of starch-carboxymethyl cellulose

This work aims to develop hydrogel films of starch and carboxymethyl cellulose (CMC) crosslinked with sodium trimetaphosphate (STMP) and to characterize some of their properties. Starch and STMP (S/T), starch and CMC (S/C), and mixed (S/T/C) films were prepared by casting. The degree of substitution, morphology, swelling degree, FTIR, mechanical properties, and sorption isotherms were studied. Reticulated samples (S/T and S/T/C) showed the same degree of substitution (0.050 ± 0.001). All films presented homogeneous morphology, but the mixed film showed greater roughness. Crosslinking increased the swelling capacity of the mixed hydrogel at pH 7, although it remained decreased concerning the S/T hydrogel. However, this property was sensitive to pH variations. The mixed film (S/T/C) showed greater mechanical resistance. The casting process was efficient to produce hydrogel films of starch/CMC crosslinked with STMP and the general results demonstrated the advantages of the mixed hydrogel.