Research updates and progress on nutritional significance of the amides I and II, alpha-helix and beta-sheet ratios, microbial protein synthesis, and steam pressure toasting condition with globar and synchrotron molecular microspectroscopic techniques with chemometrics.

This article aims to review research updates and progress on the nutritional significance of the amides I and II, the alpha-helix and beta-sheet ratios, the microbial protein synthesis, and the steam pressure toasting condition in food and feed with globar and synchrotron molecular microspectroscopic techniques plus chemometrics (both univariate and multivariate techniques). The review focused on (I) impact of the amides I and II, and the alpha-helix and beta-sheet-structure ratios in food and feeds; (II) Current research progress and update in synchrotron technique and application in feed and food molecular structure studies that are associated with nutrition delivery; (III) Impact of thermal processing- steam pressure toasting condition on feed and food; (IV). Impact of the microbial protein synthesis and methodology on feed and food; and (V). Impact on performance and production of ruminants with Faba beans.

Heat-induced amyloid-like aggregation of ß-lactoglobulin affected by glycation by α-dicarbonyl compounds in a model study.

BACKGROUND: α-Dicarbonyl compounds are widely generated in the Maillard reaction, caramelization and oil oxidation during heat treatment. These compounds can readily react with lysine and arginine residues of a protein, whereas the influence of these compounds on protein structure and quality has seldom been revealed. This study compared influence of glycation by glucose and α-dicarbonyl compounds on amyloid-like aggregation of ß-lactoglobulin (ß-LG), both fibrillation kinetics and conformation of aggregates were studied. RESULTS: Compared with glycation by glucose, the glycation by α-dicarbonyl compounds resulted in faster reduction of free amino group, sulfydryl group, and the relative content of ß-sheet secondary structure, according to the ultraviolet (UV) spectra or circular dichroism (CD) spectra results. Based on the analysis of fibrillation kinetics using thioflavin T (ThT) binding assay, the glycation by α-dicarbonyls were more efficient in suppressing the growth of fibrillar aggregates. In addition, glycation by α-dicarbonyl resulted in amorphous oligomers, which were compared with the amyloid-like aggregates in control and glucose-glycated samples, based on the transmission electron microscopy (TEM) observation. CONCLUSIONS: Glycation by α-dicarbonyl compounds induced larger decline in the ß-sheet structure of ß-LG than glycation by glucose, and thus largely suppressed the amyloid-like aggregation of ß-LG and changed the morphology of aggregates. © 2019 Society of Chemical Industry.

Ascorbic acid inhibits human insulin aggregation and protects against amyloid induced cytotoxicity.

Protein aggregation into oligomers and fibrils are associated with many human pathophysiologies. Compounds that modulate protein aggregation and interact with preformed fibrils and convert them to less toxic species, expect to serve as promising drug candidates and aid to the drug development efforts against aggregation diseases. In present study, the kinetics of amyloid fibril formation by human insulin (HI) and the anti-amyloidogenic activity of ascorbic acid (AA) were investigated by employing various spectroscopic, imaging and computational approaches. We demonstrate that ascorbic acid significantly inhibits the fibrillation of HI in a dose-dependent manner. Interestingly ascorbic acid destabilise the preformed amyloid fibrils and protects human neuroblastoma cell line (SH- SY5Y) against amyloid induced cytotoxicity. The present data signifies the role of ascorbic acid that can serve as potential molecule in preventing human insulin aggregation and associated pathophysiologies.

Structural and nanomechanical comparison of epitaxially and solution-grown amyloid ß25-35 fibrils.

Aß25-35, the fibril-forming, biologically active toxic fragment of the full-length amyloid ß-peptide also forms fibrils on mica by an epitaxial assembly mechanism. Here we investigated, by using atomic force microscopy, nanomechanical manipulation and FTIR spectroscopy, whether the epitaxially grown fibrils display structural and mechanical features similar to the ones evolving under equilibrium conditions in bulk solution. Unlike epitaxially grown fibrils, solution-grown fibrils displayed a heterogeneous morphology and an apparently helical structure. While fibril assembly in solution occurred on a time scale of hours, it appeared within a few minutes on mica surface fibrils. Both types of fibrils showed a similar plateau-like nanomechanical response characterized by the appearance of force staircases. The IR spectra of both fibril types contained an intense peak between 1620 and 1640 cm(-1), indicating that ß-sheets dominate their structure. A shift in the amide I band towards greater wave numbers in epitaxially assembled fibrils suggests that their structure is less compact than that of solution-grown fibrils. Thus, equilibrium conditions are required for a full structural compaction. Epitaxial Aß25-35 fibril assembly, while significantly accelerated, may trap the fibrils in less compact configurations. Considering that under in vivo conditions the assembly of amyloid fibrils is influenced by the presence of extracellular matrix components, the ultimate fibril structure is likely to be influenced by the features of underlying matrix elements.

A New Caged-Glutamine Derivative as a Tool To Control the Assembly of Glutamine-Containing Amyloidogenic Peptides.

We present the design, synthesis, and characterization of a novel photocaged glutamine derivative (modified on the side chain of glutamine), and describe its use in enhancing peptide stability and solubility. Our results demonstrate that this approach can be used to develop molecular switches to control the folding and ß-sheet formation of amyloidogenic peptides.

Roles of aromatic side chains and template effects of the hydrophobic cavity of a self-assembled peptide nanoarchitecture for anisotropic growth of gold nanocrystals.

Gold nanocrystals are promising as catalysts and for use in sensing/imaging systems, photonic/plasmonic devices, electronics, drug delivery systems, and for photothermal therapy due to their unique physical, chemical, and biocompatible properties. The use of various organic templates allows control of the size, shape, structure, surface modification and topology of gold nanocrystals; in particular, currently the synthesis of gold nanorods requires a cytotoxic surfactant to control morphology. To control the shape of gold nanocrystals, we previously demonstrated the de novo design and synthesis of a ß-sheet-forming nonapeptide (RU006: Ac-AIAKAXKIA-NH2, X=L-2-naphthylalanine, Nal) and the fabrication of gold nanocrystals by mixing RU006 and HAuCl4 in water. The reaction afforded ultrathin gold nanoribbons 50-100 nm wide, several nanometers high, and microns long. To understand the mechanism underlying gold nanoribbon formation by the RU006 system, we here report (i) the effects of replacement of the Nal aromatic side chain in the RU006 sequence with other aromatic moieties, (ii) the electrochemical properties of aromatic side chains in the de novo designed template peptides to estimate the redox potential and number of electrons participating in the gold crystallization process, and (iii) the stoichiometry of the RU006 system for gold nanoribbon synthesis. Interestingly, RU006 bearing a naphthalene moiety (oxidation peak potential of 1.50 V vs Ag/Ag(+)) and an analog [Ant(6)]-RU006 bearing a bulky anthracene moiety (oxidation peak potential of 1.05 V vs Ag/Ag(+)) allowed the growth of anisotropic (ribbon-like) and isotropic (round) gold nanocrystals, respectively. This trend in morphology of gold nanocrystals was attributed to spatially-arranged hydrophobic cavities sufficiently large to accommodate the gold precursor and to allow directed crystal growth driven by cross-linking reactions among the naphthalene rings. Support for this mechanism was obtained by decreasing the mole fraction of [Ant(6)]-RU006 against the total concentration of [Ant(6)]-RU006 and [Phe(6)]-RU006: absorption spectra similar to that for RU006 were obtained. Differences in the redox properties of the anthracene and naphthalene moieties scarcely affected morphology. We propose that construction of an appropriate hydrophobic cavity is important for templating gold nanocrystal architectures by peptide self-assembly. This mechanism would be applicable for developing simple, low toxicity, mild synthetic methods for constructing metallic nanomaterials for therapeutic use.

Evidence of α-helical coiled coils and ß-sheets in hornet silk.

α-Helical coiled coil and ß-sheet complexes are essential structural building elements of silk proteins produced by different species of the Hymenoptera. Beside X-ray scattering at wide and small angles we applied cryo-electron diffraction and microscopy to demonstrate the presence and the details of such structures in silk of the giant hornet Vespa mandarinia japonica. Our studies on the assembly of the fibrous silk proteins and their internal organization in relation to the primary chain structure suggest a 172 Å pitch supercoil consisting of four intertwined alanine-rich α-helical strands. The axial periodicity may adopt even multiples of the pitch value. Coiled coil motifs form the largest portion of the hornet silk structure and are aligned nearly parallel to the cocoon fiber axis in the same way as the membrane-like parts of the cocoon are molecularly orientated in the spinning direction. Supercoils were found to be associated with ß-crystals, predominantly localized in the l-serine-rich chain sequences terminating each of the four predominant silk proteins. Such ß-sheet blocks are considered resulting from transformation of random coil molecular sequences due to the action of elongational forces during the spinning process.

Structure of cytotoxic amyloid oligomers generated during disaggregation.

Amyloidosis is characterized by the abnormal accumulation of amyloid proteins. The causative proteins aggregate from monomers to oligomers and fibrils, among which some intermediate oligomers are considered as major toxins. Cytotoxic oligomers are generated not only by aggregation but also via fibril disaggregation. However, little is known about the structural characteristics and generation conditions of cytotoxic oligomers produced during disaggregation. Herein, we summarized the structural commonalities of cytotoxic oligomers formed under various disaggregation conditions, including the addition of heat shock proteins or small compounds. In vitro experimental data demonstrated the presence of high-molecular-weight oligomers (protofibrils or protofilaments) that exhibited a fibrous morphology and ß-sheet structure. Molecular dynamics simulations indicated that the distorted ß-sheet structure contributed to their metastability. The tendency of these cytotoxic oligomers to appear under mild disaggregation conditions, implied formation during the early stages of disaggregation. This review will aid researchers in exploring the characteristics of highly cytotoxic oligomers and developing drugs that target amyloid aggregates.

Ethyl 2-[(2-oxo-2H-chromen-6-yl)-oxy]acetate.

Ethyl 2-[(2-oxo-2H-chromen-6-yl)-oxy]acetate, C13H12O5, a member of the pharmacologically important class of coumarins, crystallizes in the monoclinic C2/c space group in the form of sheets, within which mol-ecules are related by inversion centers and 21 axes. Multiple C-H⋯O weak hydrogen-bonding inter-actions reinforce this pattern. The planes of these sheets are oriented in the approximate direction of the ac face diagonal. Inter-sheet inter-actions are a combination of coumarin system π-π stacking and additional C-H⋯O weak hydrogen bonds between ethyl acet-oxy groups.

Rational design of structure-based vaccines targeting misfolded alpha-synuclein conformers of Parkinson's disease and related disorders.

Synucleinopathies, including Parkinson's disease (PD), multiple system atrophy (MSA), and dementia with Lewy bodies (DLB), are neurodegenerative disorders caused by the accumulation of misfolded alpha-synuclein protein. Developing effective vaccines against synucleinopathies is challenging due to the difficulty of stimulating an immune-specific response against alpha-synuclein without causing harmful autoimmune reactions, selectively targeting only pathological forms of alpha-synuclein. Previous attempts using linear peptides and epitopes without control of the antigen structure failed in clinical trials. The immune system was unable to distinguish between native alpha-synuclein and its amyloid form. The prion domain of the fungal HET-s protein was selected as a scaffold to introduce select epitopes from the surface of alpha-synuclein fibrils. Four vaccine candidates were generated by introducing specific amino acid substitutions onto the surface of the scaffold protein. The approach successfully mimicked the stacking of the parallel in-register beta-sheet structure seen in alpha-synuclein fibrils. All vaccine candidates induced substantial levels of IgG antibodies that recognized pathological alpha-synuclein fibrils derived from a synucleinopathy mouse model. Furthermore, the antisera recognized pathological alpha-synuclein aggregates in brain lysates from patients who died from DLB, MSA, or PD, but did not recognize linear alpha-synuclein peptides. Our approach, based on the rational design of vaccines using the structure of alpha-synuclein amyloid fibrils and strict control over the exposed antigen structure used for immunization, as well as the ability to mimic aggregated alpha-synuclein, provides a promising avenue toward developing effective vaccines against alpha-synuclein fibrils.

Droplet spatial distribution of oil-based emulsion spray.

Introduction: Oil-based emulsion solution is a common pesticide formulation in agricultural spraying, and its spray characteristics are different from that of water spraying. The well understanding of its spray characteristics is the theoretical basis to improve the pesticide spraying technology. The objective of the present study is to deepen the understanding of the spray characteristics of oil-based emulsion. Method: In this paper, the spatial distribution characteristics of spray droplets of oil-based emulsion were captured visually using the high-speed photomicrography. On the basis of image processing method, the droplet size and distribution density of spray droplets at different spatial locations were analyzed quantitatively. The effects of nozzle configuration and emulsion concentration on spray structures and droplet spatial distribution were discussed. Results: Oil-based emulsion produced a special perforation atomization mechanism compared with water spray, which led to the increase of spray droplet size and distribution density. Nozzle configuration had a significant effect on oil-based emulsion spray, with the nozzle changed from ST110-01 to ST110-03 and ST110-05; the sheet lengths increased to 18 and 28 mm, respectively, whereas the volumetric median diameters increased to 51.19% and 76.00%, respectively. With emulsion concentration increased from 0.02% to 0.1% and 0.5%, the volumetric median diameters increased to 5.17% and 14.56%, respectively. Discussion: The spray droplet size of oil-based emulsion spray can be scaled by the equivalent diameter of discharge orifice of nozzles. The products of volumetric median diameters and corresponding surface tensions were nearly constant for the oil-based emulsion spray of different emulsion concentrations. It is expected that this research could provide theoretical support for improving the spraying technology of oil-based emulsion and increasing the utilization of pesticide.

Theoretical and experimental studies on the oil-based emulsion spray.

Oil-based emulsion is a common herbicide formulation in agricultural spray, and its atomization mechanism is different from that of water spray. In this paper, a theoretical model based on the characteristics of spray sheets was proposed to predict the spray droplet size for oil-based emulsion spray. An image processing method was used to measure droplet size distributions for different spray pressures and nozzle configurations, and the measured results were used to validate the theoretical model. The results show that oil-based emulsion spray is characterized by the web structure constituted by perforations. The liquid originally occupied by spray sheets eventually gathers in these web structures. The proposed theoretical model is based on the size of the nozzle exit, the angle of spray sheets, and the perforation number in the web structure, which are relatively easy to obtain. The theoretical droplet size is in inverse proportion to the square root of the perforation number in the web structure while in proportion to the square root of the area of the nozzle exit. The captured images of spray sheets and the measured droplet size distribution show consistency with the theoretical prediction. The difference between theoretical results and measured volumetric median diameter is less than 10% for different spray pressures and nozzles.

Activation of extracellular electron network in non-electroactive bacteria by Bombyx mori silk.

Extracellular electron transfer material (EETM) has increasingly attracted attentions for the enhancing effect on multiple microbial reactions. Especially, EETM is known to be essential to activate the energy network in non-electroactive bacteria. It is motivated to find out an EETM which is natural-based, environmentally friendly, and easily produced at large-scale. In this study, Bombyx mori silk is found, for the first time, to function as an EETM by using an EETM-dependent pentachlorophenol (PCP) dechlorinating anaerobic microbial culture. Subsequently, by dividing fibroin fiber into different soluble/insoluble fractions and correlating their EET functions with their structural properties based on various spectroscopic analyses, the ß-sheet configuration is suggested as an essential structure supporting the EET function of silk materials. The analyses also suggested the involvement of sulfur-containing amino acids in this function. The EET function is not degraded by boiling or acid/alkaline treatments and the material can be utilized multiple times, although it is susceptible to UV irradiation. Bombyx mori silk also enhance other microbial reactions, including Fe(III)OOH reduction, CO2 reduction to acetate, and nitrogen fixation. This discovery provides a basis for developing biotechnology for environmental remediation, global warming reduction, and biofertilizer production using Bombyx mori silk and its wastes.

Avenanthramide-C prevents amyloid formation of bovine serum albumin.

The misfolding of protein and its assembly into amyloid fibrils with a characteristic ß-sheet-rich secondary structure, cause a lot of illnesses. Polyphenols have been extensively studied as a class of amyloid inhibitors, whose effect depends on the position and number of hydroxyl groups around the flavone backbone. In this study, we used bovine serum albumin (BSA) as an amyloid model to test the anti-amyloid effects of Avenanthramide-C (Avn-C), a molecule with a long aliphatic linker between two aromatic rings. We used spectroscopy techniques like thioflavin T fluorescence and circular dichroism, to follow the ß-sheet-rich aggregates of BSA upon incubation at 68 °C. Our results demonstrated that Avn-C shows higher inhibitory effect on BSA oligomerization at micromolar concentrations, than Epigallocatechin gallate (EGCG) and Curcumin, proving for the first time, that Avn-C can serve as potential molecule in preventing protein aggregation.

A near-infrared fluorescent probe quinaldine red lights up the ß-sheet structure of amyloid proteins in mouse brain.

In this report, we describe a near-infrared fluorescent probe called quinaldine red (QR) which lights up the ß-sheet structure of amyloid fibrils. The photochemical and biophysical properties of QR along with other canonical amyloid probes in the presence of protein fibrils were investigated by using fluorescence spectroscopy, confocal fluorescent microscopy and isothermal titration calorimetry. Moreover, the binding sites and interaction mode between QR and insulin fibrils were calculated based on molecule docking. Among these amyloid probes, QR showed several advantages including strong supramolecular force, near-infrared emission, high sensitivity and resistance to bleaching. A linear response of the fluorescence intensity of QR towards fibril samples in the presence of sera was visualized in the range of 1-30 µM, with the limit of detection (LOD) of 2.31 µM. The recovery and relative standard deviation (RSD) of the proposed method for the determination of protein fibrils was 90.4%-99.2% and 3.05%-3.47%, respectively. Finally, QR can be fluorescently lighted up when meeting the aberrant protein aggregates of pathogenic mice. We recommend QR as a novel and excellent alternative tool for monitoring conformational transition of amyloid proteins.

Hot-melt Adhesive Bonding of Polyurethane/Fluorinated Polyurethane/Alkylsilane-Functionalized Graphene Nanofibrous Fabrics with Enhanced Waterproofness, Breathability, and Mechanical Properties.

Waterproof-breathable (WB) materials with outstanding waterproofness, breathability, and mechanical performance are critical in diverse consumer applications. Electrospun nanofibrous membranes with thin fiber diameters, small pore sizes, and high porosity have attracted significant attention in the WB fabric field. Hot-press treatment technology can induce the formation of inter-fiber fusion structures and hence improve the waterproofness and mechanical performance. By combining electrospinning and hot-press treatment technology, polyurethane/fluorinated polyurethane/thermoplastic polyurethane/alkylsilane-functionalized graphene (PU/FPU/TPU/FG) nanofiber WB fabric was fabricated. Subsequently, the morphologies, porous structure, hydrostatic pressure, water vapor transmission rate (WVTR), and stress-strain behavior of the nanofiber WB fabric were systematically investigated. The introduction of the hydrophobic FG sheet structure and the formation of the inter-fiber fusion structure greatly improved not only the waterproofness but also the mechanical performance of the nanofiber WB fabric. The optimized PU/FPU/TPU-50/FG-1.5 WB fabric exhibited an excellent comprehensive performance: a high hydrostatic pressure of 80.4 kPa, a modest WVTR of 7.6 kg m-2 d-1, and a robust tensile stress of 127.59 MPa, which could be used to achieve various applications. This work not only highlights the preparation of materials, but also provides a high-performance nanofiber WB fabric with huge potential application prospects in various fields.

Introduction and Fundamentals of Human Islet Amyloid Polypeptide Inhibitors.

Type 2 diabetes (T2D) is a common protein misfolding disease (PMD), and its pathogenesis is considered to be tightly associated with the aggregation of the disease-causative hIAPP (or amylin). Numerous studies have shown a possible pathological link between hIAPP aggregation and ß-cell death; thus, different-level strategies from basic research to clinical bench applications have been applied to discover and design different types of inhibitors for preventing hIAPP aggregation and toxicity. This review surveys recent and important advances of hIAPP aggregation inhibitors in the context of amyloid aggregation, toxicity, and inhibition. Many hIAPP inhibitors have been explored to exert different inhibitory functions on hIAPP aggregation via different pathways. A further overview of molecular simulations of inhibitor-hIAPP systems highlights some consensus binding sequences and structures of hIAPP for different inhibitors, which provide molecular insights into well-defined binding targets and binding-induced inhibition mechanisms for structural-based design of hIAPP inhibitors. In a broader view, while anti-aggregation inhibitors hold substantial promise in the prevention of PMDs, many challenges still remain and need to be addressed for advancing our fundamental understanding of amyloid aggregation and practical design of clinically relevant inhibitors to treat PMDs.

Self-assembly of Functional Nanostructures by Short Helical Peptide Building Blocks.

The self-assembly of short peptide building blocks into well-ordered nanostructures is a key direction in bionanotechnology. The formation of ß -sheet organizations by short peptides is well explored, leading to the development of a wide range of functional assemblies. Likewise, many natural proteinaceous materials, such as silk and amyloid fibrils, are based on ß-sheet structures. In contrast, collagen, the most abundant protein in mammals, is based on helical arrangement. Similar to ß-sheet structures, short helical peptides have been recently discovered to possess a diverse set of functionalities with the potential to fabricate artificial self-assembling materials. Here, we outline the functional roles of self-assembled nanostructures formed by short helical peptides and their potential as artificial materials. We focus on the association between self-assembled mesoscale structures and their material function and demonstrate the way by which this class of building blocks bears the potential for diverse applications, such as the future fabrication of smart devices.

On the analysis of microrheological responses of self-assembling RADA16-I peptide hydrogel.

This work aims to obtain a hydrogel based on self-assembling RADA16-I with proper rheological properties for hemostasis application. Response surface methodology (RSM) was performed to predict the gelation and stiffness of the hydrogel in different concentrations of peptide and NaCl in water and blood serum milieus. Particle tracking microrheology technique was used to evaluate Brownian motion of polystyrene particles in the peptide solutions to obtain their trajectories and measure the viscoelastic properties (G'', G″, and tan δ). Formation of gel was influenced by the concentrations of peptide and salt and their interactions. Optimum response for maximizing elastic modulus was obtained in the presence of blood serum in comparison with water. Negative effect of excess amount of NaCl was predicted by RSM model and confirmed by animal study. Circular dichroism (CD) analysis showed formation of ß-sheet secondary structure in water. On the other hand, in the presence of blood serum, tertiary structure was formed. Dimensional characterization of peptide fibers was performed by means of AFM. Peptide self-assembly in blood serum (pH around 7) which contains different ions, led to enhancing bonds between fibers, caused increasing the fiber diameter and length by 20 and 10 times, respectively. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 330-338, 2019.

Assembly of transgenic human P301S Tau is necessary for neurodegeneration in murine spinal cord.

A pathological pathway leading from soluble monomeric to insoluble filamentous Tau is characteristic of many human neurodegenerative diseases, which also exhibit dysfunction and death of brain cells. However, it is unknown how the assembly of Tau into filaments relates to cell loss. To study this, we first used a mouse line transgenic for full-length human mutant P301S Tau to investigate the temporal relationship between Tau assembly into filaments, assessed using anti-Tau antibody AT100, and motor neuron numbers, in the lumbar spinal cord. AT100 immunoreactivity preceded nerve cell loss. Murine Tau did not contribute significantly to either Tau aggregation or neurodegeneration. To further study the relevance of filament formation for neurodegeneration, we deleted hexapeptides 275VQIINK280 and 306VQIVYK311, either singly or in combination, from human 0N4R Tau with the P301S mutation. These hexapeptides are essential for the assembly of Tau into filaments. Homozygous mice transgenic for P301S Tau with the hexapeptide deletions, which expressed Tau at a similar level to the heterozygous line transgenic for P301S Tau, had a normal lifespan, unlike mice from the P301S Tau line. The latter had significant levels of sarkosyl-insoluble Tau in brain and spinal cord, and exhibited neurodegeneration. Mice transgenic for P301S Tau with the hexapeptide deletions failed to show significant levels of sarkosyl-insoluble Tau or neurodegeneration. Recombinant P301S Tau with the hexapeptide deletions failed to form ß-sheet structure and filaments following incubation with heparin. Taken together, we conclude that ß-sheet assembly of human P301S Tau is necessary for neurodegeneration in transgenic mice.