Determination of the Residual Life of the Spraying Boom of a Field Sprinkler in the Maneuvering Loading Mode

Materials Science - We propose a computational model for the evaluation of the residual service life of spraying booms of the field sprinklers in the maneuvering loading mode (stationary...     

Tyrp1 Mutant Variants Associated with OCA3: Computational Characterization of Protein Stability and Ligand Binding

Oculocutaneous albinism type 3 (OCA3) is an autosomal recessive disorder caused by mutations in the TYRP1 gene. Tyrosinase-related protein 1 (Tyrp1) is involved in eumelanin synthesis, catalyzing the oxidation of 5,6-dihydroxyindole-2-carboxylic acid oxidase (DHICA) to 5,6-indolequinone-2-carboxylic acid (IQCA). Here, for the first time, four OCA3-causing mutations of Tyrp1, C30R, H215Y, D308N, and R326H, were investigated computationally to understand Tyrp1 protein stability and catalytic activity. Using the Tyrp1 crystal structure (PDB:5M8L), global mutagenesis was conducted to evaluate mutant protein stability. Consistent with the foldability parameter, C30R and H215Y should exhibit greater instability, and two other mutants, D308N and R326H, are expected to keep a native conformation. SDS-PAGE and Western blot analysis of the purified recombinant proteins confirmed that the foldability parameter correctly predicted the effect of mutations critical for protein stability. Further, the mutant variant structures were built and simulated for 100 ns to generate free energy landscapes and perform docking experiments. Free energy landscapes formed by Y362, N378, and T391 indicate that the binding clefts of C30R and H215Y mutants are larger than the wild-type Tyrp1. In docking simulations, the hydrogen bond and salt bridge interactions that stabilize DHICA in the active site remain similar among Tyrp1, D308N, and R326H. However, the strengths of these interactions and stability of the docked ligand may decrease proportionally to mutation severity due to the larger and less well-defined natures of the binding clefts in mutants. Mutational perturbations in mutants that are not unfolded may result in allosteric alterations to the active site, reducing the stability of protein-ligand interactions.     

Assessment of subcritical crack growth in hydrogen-containing environment by the parameters of acoustic emission signals

Estimation models (differential equations, initial and final conditions) for determining the crack propagation kinetics in hydrogen-containing environments using the acoustic emission (AE) signal parameters are proposed. The formulation of these models is based on the main ideals of the AE method, dependence between the crack increment area and a sum of AE-signals amplitude, main criteria of fracture mechanics and laws of thermodynamics.     

Tyrosinase Nanoparticles: Understanding the Melanogenesis Pathway by Isolating the Products of Tyrosinase Enzymatic Reaction

Human Tyrosinase (Tyr) is the rate-limiting enzyme of the melanogenesis pathway. Tyr catalyzes the oxidation of the substrate L-DOPA into dopachrome and melanin. Currently, the characterization of dopachrome-related products is difficult due to the absence of a simple way to partition dopachrome from protein fraction. Here, we immobilize catalytically pure recombinant human Tyr domain (residues 19–469) containing 6xHis tag to Ni-loaded magnetic beads (MB). Transmission electron microscopy revealed Tyr-MB were within limits of 168.2 ± 24.4 nm while the dark-brown melanin images showed single and polymerized melanin with a diameter of 121.4 ± 18.1 nm. Using Hill kinetics, we show that Tyr-MB has a catalytic activity similar to that of intact Tyr. The diphenol oxidase reactions of L-DOPA show an increase of dopachrome formation with the number of MB and with temperature. At 50 °C, Tyr-MB shows some residual catalytic activity suggesting that the immobilized Tyr has increased protein stability. In contrast, under 37 °C, the dopachrome product, which is isolated from Tyr-MB particles, shows that dopachrome has an orange-brown color that is different from the color of the mixture of L-DOPA, Tyr, and dopachrome. In the future, Tyr-MB could be used for large-scale productions of dopachrome and melanin-related products and finding a treatment for oculocutaneous albinism-inherited diseases.     

Determination of the Period of Subcritical Growth of Small Plane High-Temperature Creep Cracks in Structural Elements

Materials Science - We formulate a computation model (differential equation with initial and final conditions) for the determination of the period of subcritical growth of small plane...     

Theoretical Foundations of the Method of Acoustic Emission for the Diagnostics of Delayed Fracture of Materials

Materials Science - We present a survey of the results of theoretical investigations of the method of acoustic emission used for the determination of the limit equilibrium of materials with cracks,...     

Computational Model for the Evaluation of the Service Life Of Fiber-Reinforced Concrete Structures Under Long-Term Static Loading

Materials Science - We propose a computational model for the evaluation of the service life of fiber-reinforced concrete structural elements subjected to long-term tension. The model is based on...     

Dynamic control of flexible manufacturing systems

Flexible manufacturing systems (FMSs) are a relatively new technological and organisational approach to helping companies respond to real-time marketing conditions for their production. Under a proposal of the National Bureau of Standards the FMSs are subdivided into virtual manufacturing cells in a dynamic manner, on the basis of group technology.A method of dynamic optimisation for the design of manufacturing processes, capacity balancing and checking, and also production scheduling or rescheduling in virtual manufacturing cells is described. It can be used during real-time production control in FMSs.     

Synaptosomal uptake of alpha-ketoglutarate and glutamine in thioacetamide-induced hepatic encephalopathy in rats

The Hospital Anxiety and Depression Scale (HAD) was evaluated in a Swedish population sample. The purpose of the study was to compare the HAD with the Beck Depression Inventory (BDI) and Spielberger's State Trait Anxiety Inventory (STAI). A secondary aim was to examine the factor structure of the HAD. The results indicated that the factor structure was quite strong, consistently showing two factors in the whole sample as well as in different subsamples. The correlations between the total HAD scale and BDI and STAI, respectively, were stronger than those obtained using the different subscales of the HAD (the anxiety and depression subscales). As expected, there was also a stronger correlation between the HAD and the non-physical items of the BDI. It was somewhat surprising that the factor analyses were consistently extracting two factors, 'depression' and 'anxiety', while on the other hand both BDI and STAI tended to correlate more strongly with the total HAD score than with the specific depression and anxiety HAD subscales. Nevertheless, the HAD appeared to be (as was indeed originally intended) a useful clinical indicator of the possibility of depression and clinical anxiety.     

Characterization of Temperature-Dependent Kinetics of Oculocutaneous Albinism-Causing Mutants of Tyrosinase

Human tyrosinase (Tyr) is a glycoenzyme that catalyzes the first and rate-limiting step in melanin production, and its gene (TYR) is mutated in many cases of oculocutaneous albinism type 1 (OCA1). The mechanisms by which individual mutations contribute to the diverse pigmentation phenotype in patients with OCA1 have only began to be examined and remain to be delineated. Here, we analyze the temperature-dependent kinetics of wild-type Tyr (WT) and two OCA1B mutant variants (R422Q and P406L) using Michaelis–Menten and Van’t Hoff analyses. Recombinant truncated human Tyr proteins (residues 19–469) were produced in the whole insect Trichoplusia Ni larvae. Proteins were purified by a combination of affinity and size-exclusion chromatography. The temperature dependence of diphenol oxidase protein activities and kinetic parameters were measured by dopachrome absorption. Using the same experimental conditions, computational simulations were performed to assess the temperature-dependent association of L-DOPA and Tyr. Our results revealed, for the first time, that the association of L-DOPA with R422Q and P406L followed by dopachrome formation is a complex reaction supported by enthalpy and entropy forces. We show that the WT has a higher turnover number as compared with both R422Q and P406L. Elucidating the kinetics and thermodynamics of mutant variants of Tyr in OCA1B helps to understand the mechanisms by which they lower Tyr catalytic activity and to discover novel therapies for patients.