A Complexed Crystal Structure of a Single-Stranded DNA-Binding Protein with Quercetin and the Structural Basis of Flavonol Inhibition Specificity

Single-stranded DNA (ssDNA)-binding protein (SSB) plays a crucial role in DNA replication, repair, and recombination as well as replication fork restarts. SSB is essential for cell survival and, thus, is an attractive target for potential antipathogen chemotherapy. Whether naturally occurring products can inhibit SSB remains unknown. In this study, the effect of the flavonols myricetin, quercetin, kaempferol, and galangin on the inhibition of Pseudomonas aeruginosa SSB (PaSSB) was investigated. Furthermore, SSB was identified as a novel quercetin-binding protein. Through an electrophoretic mobility shift analysis, myricetin could inhibit the ssDNA binding activity of PaSSB with an IC50 of 2.8 ± 0.4 μM. The effect of quercetin, kaempferol, and galangin was insignificant. To elucidate the flavonol inhibition specificity, the crystal structure of PaSSB complexed with the non-inhibitor quercetin was solved using the molecular replacement method at a resolution of 2.3 Å (PDB entry 7VUM) and compared with a structure with the inhibitor myricetin (PDB entry 5YUN). Although myricetin and quercetin bound PaSSB at a similar site, their binding poses were different. Compared with myricetin, the aromatic ring of quercetin shifted by a distance of 4.9 Å and an angle of 31o for hydrogen bonding to the side chain of Asn108 in PaSSB. In addition, myricetin occupied and interacted with the ssDNA binding sites Lys7 and Glu80 in PaSSB whereas quercetin did not. This result might explain why myricetin could, but quercetin could not, strongly inhibit PaSSB. This molecular evidence reveals the flavonol inhibition specificity and also extends the interactomes of the natural anticancer products myricetin and quercetin to include the OB-fold protein SSB.

Design and Development of Hybrid Converter for Marine Applications

Recently, there has been an increase in the growth and advancement of electric propulsion in marine electrical drives. A maximum amount of energy is utilized by ships for propulsion drives. To be aware of it and develop an optimized structure to improve the effectiveness of the propulsion system with power consumption is necessary. The proposed paper aims to develop a model and perform functional analysis as per the above understanding and requirements. The factors considered include greenhouse gas emissions, CO2 emissions, environmental aspects, and the availability of non-renewable resources, which leads to the introduction of renewable energy as a replacement method of power generation. For this work, two different renewable sources, such as solar and wind energy, were chosen. The combination of these two resources can manipulate the voltage and satisfy the load in a desirable way. For voltage improvement, a high gain converter with a minimal number of active and passive components is selected. This system adopts a storage system to meet the needs in the future. The inverter switches are controlled by the recommended control algorithm, which can balance and provide adequate power towards the drive by a feedback control loop. The speed of propulsion in the drive is adjusted by the induction motor coupled with the propeller. The analytical study of the proposed system is carried out in MATLAB software. The simulation study revealed the effectiveness of this modern optimization technique.

Sequence Analysis and Preliminary X-ray Crystallographic Analysis of an Acetylesterase (LgEstI) from Lactococcus garvieae

A gene encoding LgEstI was cloned from a bacterial fish pathogen, Lactococcus garvieae. Sequence and bioinformatic analysis revealed that LgEstI is close to the acetyl esterase family and had maximum similarity to a hydrolase (UniProt: Q5UQ83) from Acanthamoeba polyphaga mimivirus (APMV). Here, we present the results of LgEstI overexpression and purification, and its preliminary X-ray crystallographic analysis. The wild-type LgEstI protein was overexpressed in Escherichia coli, and its enzymatic activity was tested using p-nitrophenyl of varying lengths. LgEstI protein exhibited higher esterase activity toward p-nitrophenyl acetate. To better understand the mechanism underlying LgEstI activity and subject it to protein engineering, we determined the high-resolution crystal structure of LgEstI. First, the wild-type LgEstI protein was crystallized in 0.1 M Tris-HCl buffer (pH 7.1), 0.2 M calcium acetate hydrate, and 19% (w/v) PEG 3000, and the native X-ray diffraction dataset was collected up to 2.0 Å resolution. The crystal structure was successfully determined using a molecular replacement method, and structure refinement and model building are underway. The upcoming complete structural information of LgEstI may elucidate the substrate-binding mechanism and provide novel strategies for subjecting LgEstI to protein engineering.

Investigation of Individual Cells Replacement Concept in Lithium-Ion Battery Packs with Analysis on Economic Feasibility and Pack Design Requirements

The optimization of lithium-ion (Li-ion) battery pack usage has become essential due to the increasing demand for Li-ion batteries. Since degradation in Li-ion batteries is inevitable, there has been some effort recently on research to maximize the utilization of Li-ion battery cells in the pack. Some promising concepts include reconfigurable battery packs and cell replacement to limit the negative impact of early-degraded cells on the entire pack. This paper used a simulation framework, based on a cell voltage model and a degradation model, to study the feasibility and benefits of the cell replacement concept. The simulation conducted in MATLAB involves generating and varying Li-ion cells in the packs stochastically and simulating the life of the cells as well as the packs until they reach their end-of-life stage. It was found that the cell replacement method can increase the total number of cycles of the battery packs, effectively prolonging the lifespan of the packs. It is also determined that this approach can be more economically beneficial than the current approach of simple pack replacement. For the cell replacement concept to be practical, two main design criteria should be satisfied including individual cell monitoring and easy accessibility to cells at failure stage.

Ag–Au Core–Shell Triangular Nanoprisms for Improving p-g-C3N4 Photocatalytic Hydrogen Production

Ag–Au core–shell triangular nanoprisms (Ag@Au TNPs) have aroused extensive research interest in the field of hydrogen evolution reaction (HER) due to their strong plasmon effect and stability. Here, Ag@Au TNPs were fabricated by the galvanic-free replacement method. Then, we loaded them on protonated g-C3N4 nanoprisms (P–CN) by the electrostatic self-assembly method as an efficient plasmonic photocatalyst for HER. The hydrogen production rate of Ag@Au TNPs/P–CN (4.52 mmol/g/h) is 4.1 times higher than that of P–CN (1.11 mmol/g/h) under simulated sunlight irradiation, making it the most competitive material for water splitting. The formed Schottky junction helps to trap the hot electrons generated from Ag@Au TNPs, and the well-preserved tips of the Ag@Au TNPs can effectively generate an electromagnetic field to inhibit the photogenerated electron–holes pairs recombination. This study suggests that the rational design of Ag@Au TNPs by the galvanic-free replacement method is an effective co-catalyst for HER and boosting the additional combination of plasmonic metals and catalyst metals for the enhancement to HER.

Imaging-Guided Bioreactor for De-Epithelialization and Long-Term Cultivation of Ex Vivo Rat Trachea

Recent synergistic advances in organ-on-chip and tissue engineering technologies offer opportunities to create in vitro-grown tissue or organ constructs that can faithfully recapitulate their in vivo counterparts. Such in vitro tissue or organ constructs can be utilized in multiple applications, including rapid drug screening, high-fidelity disease modeling, and precision medicine. Here, we report an imaging-guided bioreactor that allows in situ monitoring of the lumen of ex vivo airway tissues during controlled in vitro tissue manipulation and cultivation of isolated rat trachea. Using this platform, we demonstrated selective removal of the rat tracheal epithelium (i.e., de-epithelialization) without disrupting the underlying subepithelial cells and extracellular matrix. Through different tissue evaluation assays, such as immunofluorescent staining, DNA/protein quantification, and electron beam microscopy, we showed that the epithelium of the tracheal lumen can be effectively removed with negligible disruption in the underlying tissue layers, such as cartilage and blood vessel. Notably, using a custom-built micro-optical imaging device integrated with the bioreactor, the trachea lumen was visualized at the cellular level in real time, and removal of the endogenous epithelium and distribution of locally delivered exogenous cells were demonstrated in situ. Moreover, the de-epithelialized trachea supported on the bioreactor allowed attachment and growth of exogenous cells seeded topically on its denuded tissue surface. Collectively, the results suggest that our imaging-enabled rat trachea bioreactor and selective cell replacement method can facilitate creating of bioengineered in vitro airway tissue that can be used in different biomedical applications.

Sustainable ground improvement method using encapsulated polypropylene (PP) column reinforcement

This study investigates the effectiveness of encapsulated polypropylene (PP) column in enhancing the undrained shear strength of kaolin (soft clay). The usage of PP in treating problematic soil is a more sustainable and cost-effective alternative compared to other materials. The installation of granular column can be done by using vibro-replacement method. Several geotechnical tests to determine the properties of materials were conducted. The shear strength of treated kaolin sample was examined by using Unconfined Compression Test (UCT). There are seven (7) batches of soil sample in total which included a control sample, three (3) batches of 14 mm and three (3) batches of 20 mm diameter PP column. Different diameters of PP column were examined with 60 mm, 80 mm and 100 mm height, respectively with soil sample of 50 mm in diameter and 100 mm in height. The shear strength improvement of kaolin is 33.82%, 46.51%, and 49.88% when implanted with a PP column with a 7.84 area replacement ratio and 0.6, 0.8 and 1.0 penetration ratio. The soft soil treated using 16.00 area replacement ratio with 0.6, 0.8 and 1.0 penetration ratio has a shear strength increment of 25.22%, 33.39% and 37.59% respectively. In short, the shear strength improvement of the kaolin clay depends on the parameter of the PP column used to reinforce the sample.

Sea Bass Primary Cultures versus RTgill-W1 Cell Line: Influence of Cell Model on the Sensitivity to Nanoparticles

Determination of acute toxicity to vertebrates in aquatic environments is mainly performed following OECD test guideline 203, requiring the use of a large number of fish and with mortality as endpoint. This test is also used to determine toxicity of nanomaterials in aquatic environments. Since a replacement method for animal testing in nanotoxicity studies is desirable, the feasibility of fish primary cultures or cell lines as a model for nanotoxicity screenings is investigated here. Dicentrarchus labrax primary cultures and RTgill-W1 cell line were exposed to several concentrations (0.1 to 200 ug/mL) of different nanoparticles (TiO2, polystyrene and silver), and cytotoxicity, metabolic activity and reactive oxygen species formation were investigated after 24 and 48 h of exposure. Protein corona as amount of protein bound, as well as the influence of surface modification (-COOH, -NH2), exposure media (Leibovitz’s L15 or seawater), weathering and cell type were the experimental variables included to test their influence on the results of the assays. Data from all scenarios was split based on the significance each experimental variable had in the result of the cytotoxicity tests, in an exploratory approach that allows for better understanding of the determining factors affecting toxicity. Data shows that more variables significantly influenced the outcome of toxicity tests when the primary cultures were exposed to the different nanoparticles. Toxicity tests performed in RTgill-W1 were influenced only by exposure time and nanoparticle concentration. The whole data set was integrated in a biological response index to show the overall impact of nanoparticle exposures.