Association Mechanism and Conformational Changes in Trypsin on Its Interaction with Atrazine: A Multi- Spectroscopic and Biochemical Study with Computational Approach

Atrazine (ATR) is a herbicide globally used to eliminate undesired weeds. Herbicide usage leads to various adverse effects on human health and the environment. The primary source of herbicides in humans is the food laced with the herbicides. The ATR binding to trypsin (TYP) was investigated in this study to explore its binding potential and toxicity. In vitro interaction of ATR with TYP was studied using multi-spectroscopic methods, molecular docking, and enzyme kinetics to explore the mechanism of binding for the TYP-ATR system. The TYP-ATR complex revealed binding constants (10³ M⁻¹), suggesting a moderate binding. The free energy for the TYP-ATR complexes was negative, suggesting a spontaneous interaction. Thermodynamic parameters enthalpy (ΔH) and entropy (ΔS) obtained positive values for the TYP-ATR system suggesting hydrophobic interactions in the binding process. Micro-environmental and conformational changes in TYP molecules were induced on interaction with ATR. Reduced catalytic activity of TYP was observed after interaction with ATR owing to the changes in the secondary structure of the TYP.     

Generalized Approach towards Secretion-Based Protein Production via Neutralization of Secretion-Preventing Cationic Substrate Residues

Many heterologous proteins can be secreted by bacterial ATP-binding cassette (ABC) transporters, provided that they are fused with the C-terminal signal sequence, but some proteins are not secretable even though they carry the right signal sequence. The invention of a method to secrete these non-secretable proteins would be valuable both for understanding the secretory physiology of ABC transporters and for industrial applications. Herein, we postulate that cationic “supercharged” regions within the target substrate protein block the secretion by ABC transporters. We also suggest that the secretion of such substrate proteins can be rescued by neutralizing those cationic supercharged regions via structure-preserving point mutageneses. Surface-protruding, non-structural cationic amino acids within the cationic supercharged regions were replaced by anionic or neutral hydrophilic amino acids, reducing the cationic charge density. The examples of rescued secretions we provide include the spike protein of SARS-CoV-2, glutathione-S-transferase, streptavidin, lipase, tyrosinase, cutinase, growth factors, etc. In summary, our study provides a method to predict the secretability and a tool to rescue the secretion by correcting the secretion-blocking regions, making a significant step in understanding the physiological properties of ABC transporter-dependent protein secretion and laying the foundation for the development of a secretion-based protein-producing platform.     

VLDL receptor mediates the uptake of human chylomicron remnants in vitro

The VLDL receptor has been described as a new member of the LDL receptor supergene family that specifically binds VLDL in vitro via apolipoprotein E and lipoprotein lipase. Both apolipoprotein E and lipoprotein lipase are constituents of chylomicron remnants, another triglyceride-rich lipoprotein which has been proposed as a physiological ligand for the VLDL receptor. We used human chylomicron remnants to study their uptake into LDL, receptor-deficient Chinese hamster ovary cells overexpressing the human VLDL receptor. The uptake into these cells was compared to that into cells transfected with an empty transfection vector. Human chylomicron remnants were produced in vitro by hydrolysis with lipoprotein lipase, and were labeled with 125I. The uptake of these remnants into the cells overexpressing the VLDL receptor was found to be about 3-fold higher than the uptake into the control cells. The addition of a surplus of either apolipoprotein E or inactivated lipoprotein lipase to the remnants led to an increase in particle uptake. The chylomicron remnant uptake was inhibited by addition of the 39 kDa receptor associated protein These in vitro experiments strongly support the idea that the VLDL receptor is a physiological receptor for chylomicron remnants. The increase of receptor-mediated uptake induced by the addition of apoE or lipoprotein lipase underlines the role of these two proteins in this process.     

Optimization of abrasive water jet cutting of ductile materials

Full factorial design of experiments was developed in order to investigate the effects of jet pressure, abrasive mixing rate, cutting feed, and plate thickness upon three response variables, surface finish of cutting wear zone, percentage proportion of striation free area, and maximum width of cut. The set of sixteen experiments was performed on each of the following two ductile materials: AISI 4340 (high strength low alloy steel, hardened to 49HRc) and Aluminum 2219. Analysis of Variance (ANOVA) was performed on experimental data in order to determine the significance of effects of different parameters on the performance measures. It was found that cutting feed and thickness were highly influential parameters, while abrasive mixing rate is influential upon surface roughness only. Strong interaction was found between jet pressure and workpiece material. Multi-criteria numerical optimization was performed in order to simultaneously maximize/minimize different combinations of performance measures.     

The influence of the material toughness and the state of stress on fracture of large scale specimens

The influence of the material toughness on fracture behaviour of large scale tensile specimens is being described in this report. Selected large scale specimens show that the amount of stable crack growth prior to instability is affected by the constraint in the specimen. The constraint depends on the specimen geometry, the specimen size, and in the case of elastic plastic material behaviour on the ductility of the material. A possibility of estimating these influences is presented by quantifying the constraint with the multi-axiality quotient q.     

A computer program for internal conversion coefficients and particle parameters

Title of program: CATAR Catalogue number: ABGP Program obtainable from: CPC Program Library, Queen's University of Belfast, N. Ireland (see application form in this issue) Computer: UNIVAC 1108; Installation: Sandoz A.G., Basel, Switzerland Operating system: EXEC 8 Programming language used: FORTRAN IV High speed store required: 26K words No. of bits in a word: 36 Overlay structure: None Number of magnetic tapes required: None Other peripherals used: Card reader, line printer Number of cards in combined program and test deck: 3136 Card punching code: BCD     

Thiogenistein—Antioxidant Chemistry,Antitumor Activity,and Structure Elucidation of New Oxidation Products

Isoflavonoids such as genistein (GE) are well known antioxidants. The predictive biological activity of structurally new compounds such as thiogenistein (TGE)–a new analogue of GE–becomes an interesting way to design new drug candidates with promising properties. Two oxidation strategies were used to characterize TGE oxidation products: the first in solution and the second on the 2D surface of the Au electrode as a self-assembling TGE monolayer. The structure elucidation of products generated by different oxidation strategies was performed. The electrospray ionization mass spectrometry (ESI-MS) was used for identifying the product of electrochemical and hydrogen peroxide oxidation in the solution. Fourier transform infrared spectroscopy (FT-IR) with the ATR mode was used to identify a product after hydrogen peroxide treatment of TGE on the 2D surface. The density functional theory was used to support the experimental results for the estimation of antioxidant activity of TGE as well as for the molecular modeling of oxidation products. The biological studies were performed simultaneously to assess the suitability of TGE for antioxidant and antitumor properties. It was found that TGE was characterized by a high cytotoxic activity toward human breast cancer cells. The research was also carried out on mice macrophages, disclosing that TGE neutralized the production of the LPS-induced reactive oxygen species (ROS) and exhibits ABTS (2,2′-azino-bis-3-(ethylbenzothiazoline-6-sulphonic acid) radical scavenging ability. In the presented study, we identified the main oxidation products of TGE generated under different environmental conditions. The electroactive centers of TGE were identified and its oxidation mechanisms were proposed. TGE redox properties can be related to its various pharmacological activities. Our new thiolated analogue of genistein neutralizes the LPS-induced ROS production better than GE. Additionally, TGE shows a high cytotoxic activity against human breast cancer cells. The viability of MCF-7 (estrogen-positive cells) drops two times after a 72-h incubation with 12.5 μM TGE (viability 53.86%) compared to genistein (viability 94.46%).     

Si Doping of GaN in Hydride Vapor-Phase Epitaxy

Growth of GaN boules by hydride vapor-phase epitaxy (HVPE) is very attractive for fabrication of GaN substrates. Use of dichlorosilane as a source for Si doping of bulk GaN is investigated. It is shown that no tensile strain is incorporated into mm-thick, Si-doped GaN layers on sapphire substrates if the threading dislocation density is previously reduced to 2.5 × 10⁷ cm^?2 or below. High-quality GaN layers with electron densities up to 1.5 × 10¹⁹ cm^?3 have been achieved, and an upper limit of about 4 × 10¹⁹ cm^?3 for Si doping of GaN boules was deduced considering the evolution of dislocations with thickness. A 2-inch, Si-doped GaN crystal with length exceeding 6 mm and targeted Si doping of about 1 × 10¹⁸ cm^?3 is demonstrated.     

Probing the electromagnetic field distribution within a metallic nanodisk

A Co nanolayer is used as a local probe to evaluate the vertical inhomogeneous distribution of the electromagnetic (EM) field within a resonant metallic nanodisk. Taking advantage of the direct relation between the magneto-optical activity and the electromagnetic field intensity in the Co layer, it is shown that the nonuniform EM distribution within the nanodisk under plasmon resonant conditions has maximum values close to the upper and lower flat faces, and a minimum value in the middle.