Confocal Laser Scanning Microscopy Technique Research Articles

Paracoccidioides spp.: the structural characterization of extracellular matrix, expression of glucan synthesis and associated genes and adhesins during biofilm formation.

The genus Paracoccidioides includes Paracoccidioides lutzii and the Paracoccidioides brasiliensis complex, which comprises four phylogenetic species. A key feature distinguishing planktonic growth from biofilm is the presence of a 3D extracellular matrix (ECM). Therefore, in this study, we analyzed biofilm formation in different species of Paracoccidioides yeast phase, characterized the structural elements of the matrix of P. brasiliensis (Pb18), P. lutzii (Pl01 and 8334) and P. restrepiensis (339 and 192) and evaluated the expression of glucan genes, according to the stage of biofilm evolution for P. brasiliensis. The strains were cultivated in planktonic and biofilm form for 24-144 h. The fungi biomass and metabolic activity were determined by crystal violet and tetrazolium salt reduction (XTT) tests and colony-forming unit (CFU) by plating. The biofilm structure was designed using scanning electron microscopy and confocal laser scanning microscopy techniques. The extracellular matrix of P. brasiliensis and P. lutzii biofilms was extracted by sonication, and polysaccharides, proteins, and extracellular DNA (eDNA) were quantified. The RNA was extracted with the Trizol® reagent and quantified; then, the cDNA was synthesized to analyze the enolase expression, 14-3-3, FKS1, AGS1, GEL3, and KRE6 genes by real-time PCR. All strains of Paracoccidioides studied form a biofilm with more significant metabolic activity and biomass values in 144 h. The extracellular matrix of P. brasiliensis and P. lutzii had a higher content of polysaccharides in their composition, followed by proteins and eDNA in smaller quantities. The P. brasiliensis biofilm kinetics of formation showed greater expression of genes related to glucan's synthesis and its delivery to the external environment in addition adhesins during the biofilm's adhesion, initiation, and maturation. The GEL3 and enolase genes increased in expression within 24 h and during the biofilm maturation period, there was an increase in 14-3-3, AGS1, and FKS1. Furthermore, at 144 h, there was a decrease in KRE6 expression and an increase in GEL3. This study highlights the potential for biofilm formation for three species of Paracoccidioides and the main components of the extracellular matrix that can contribute to a better understanding of biofilm organization.

Prior roasting of wheat impacts on the functionality of flour prepared from it: Part 1. Wheat flour viscosifying properties

AbstractBackground and ObjectivesDry thermal treatment of wheat modifies the properties of wheat and of flour prepared from it. The objective of this study was to investigate the effect of wheat roasting on microstructural properties of wheat grains using confocal laser scanning, X‐ray diffraction (XRD), and scanning electron microscopy (SEM) techniques. The impact of roasting on the damaged starch contents, the solvent retention capacity profiles, and the pasting properties of the resulting flour prepared from roasted wheat were also studied.FindingsResponse surface models predicted wheat roasting at 115°C and 65 Hz (165 s) to enhance flour viscosifying properties. Roasting before milling resulted in grain puffing, slight damage to the starch granule surface, and destruction of the protein matrix, as observed using SEM. XRD analysis revealed that amylose–lipid complexation had occurred. As a result of roasting, the peak, hot paste, and final viscosities of flour from roasted wheat were significantly (p ≤ .05) higher than those of flour produced from control wheat.ConclusionsRoasting induces structural changes in wheat. Flour produced from roasted wheat can be used to produce gel structures with increased viscosities.Significance and NoveltyDry thermal treatment improves the viscosifying properties of flour milled from roasted wheat.

Mechanistic enhancement of emulsification function in zein/pectin complex nanoparticles by short linear glucan

The objective of this research was to create stable Pickering emulsions (PEs) by manipulating the structure of the interface through the control of interfacial properties, self-assembly, and packing characteristics of zein particles, utilizing the interaction between short linear glucan (SLG) and pectin. The scientists successfully synthesized ternary complex particles designated as SZPPs (abbreviated for SLG/Zein/Pectin), displaying three-phase contact angles ranging from 83.14° to 87.32°. SZPPs were effectively anchored at the oil-water interface, resulting in a robust and well-organized arrangement, as confirmed by the combination of Langmuir−Blodgett and Scanning Electron Microscope (LB-SEM) and confocal laser scanning microscopy techniques. This arrangement led to the formation of a permeating three-dimensional network of oil droplets, which greatly contributed to the creation of stable PEs. These PEs exhibited unique properties such as viscoelasticity, remarkable thixotropy, and excellent long-term storage stability. One noteworthy advantage of these engineered PEs was their ability to shield curcumin from degradation caused by UV radiation, thereby enhancing the oxidative stability of the emulsions. This protective effect ensures that the curcumin remains intact and active over time. Considering these advantageous properties, these engineered PEs hold great promise for various applications in the food and pharmaceutical industries.

Bone Hierarchical Structure: Heterogeneity and Uniformity

AbstractBone has a complex hierarchical structure with structural integration from nm to cm. The understanding of bone structure is developing rapidly due to improvements in available methodologies that allow unravelling structures across several length scales. These methods include advances in electron microscopy, in particular, focused ion beam scanning electron microscopy (FIB‐SEM), confocal laser scanning microscopy techniques, X‐ray imaging, X‐ray diffraction tomography (XRD‐CT), and tensor tomography (small angle X‐ray scattering tensor tomgraphy, SAXS‐TT and wide angle X‐ray scattering tensor tomgraphy, WAXS‐TT). Special emphasis is placed on the latter X‐ray techniques that are emerging into powerful tools. Through a review of selected recent results on the structure of the bone matrix as well as the lacuno‐canalicular network housing the osteocyte cells of bone, it is proposed that bone is more heterogeneous than typically described and that local variation in composition and crystallography may play a significant role in bone biology in health and disease.

Boring bryozoans: an investigation into the endolithic bryozoan family Penetrantiidae

An endolithic lifestyle in mineralized substrates has evolved multiple times in various phyla including Bryozoa. The family Penetrantiidae includes one genus with ten extant and two fossil species. They predominantly colonize the shells of molluscs and establish colonies by chemical dissolution of calcium carbonate. Based on several morphological characters, they were described to be either cheilostome or ctenostome bryozoans. For more than 40 years, neither the characters of species identity and systematics nor the problem of their phylogeny was approached. Consequently, the aim of this study is to reevaluate species identities and the systematic position of the genus Penetrantia by analyzing at least six different species from eight regions with the aid of modern methods such as confocal laser scanning microscopy and 3D-reconstruction techniques. This study demonstrates that the musculature associated with the operculum and brood chamber shows significant differences from the cheilostome counterparts and seems to have evolved independently. Together with the presence of other ctenostome-like features such as true polymorphic stolons and uncalcified body wall, this finding supports a ctenostome affinity. Operculum morphology reveals many new species-specific characters, which, together with information about gonozooid morphology, tentacle number, and zooid size ranges, will enhance species identification. It also revealed a probable new species in Japan as well as potential cryptic species in France and New Zealand. In addition, this study increases the known distribution range of the family and its substrate diversity. Altogether, the new information collated here provides the basis for future work on a neglected taxon.

Mangiferin-Enriched Mn-Hydroxyapatite Coupled with β-TCP Scaffolds Simultaneously Exhibit Osteogenicity and Anti-Bacterial Efficacy.

Biphasic calcium phosphate (BCP) containing β-tricalcium phosphate and manganese (Mn)-substituted hydroxyapatite (HAP) was synthesized. Biomedical scaffolds were prepared using this synthesized powder on a sacrificial polyurethane sponge template after the incorporation of mangiferin (MAN). Mn was substituted at a concentration of 5% and 10% in HAP to examine the efficacy of Mn at various concentrations. The phase analysis of the as-formed BCP scaffold was carried out by X-ray diffraction analysis, while the qualitative observation of morphology and the osteoblast cell differentiation were carried out by scanning electron microscopy and confocal laser scanning microscopy techniques. Gene expressions of osteocalcin, collagen 1, and RUNX2 were carried out using qRT-PCR analyses. Significantly higher (p < 0.05) levels of ALP activity were observed with extended osteoblast induction on the mangiferin-incorporated BCP scaffolds. After characterization of the specimens, it was found that the scaffolds with 10% Mn-incorporated BCP with mangiferin showed better osteogenicity and simultaneously the same scaffolds exhibited higher anti-bacterial properties as observed from the bacterial viability test. This study was carried out to evaluate the efficacy of Mn and MAN in BCP for osteogenicity and antibacterial action.

Nanoparticle Coatings on Glass Surfaces to Prevent Pseudomonas fluorescens AR 11 Biofilm Formation.

Microbial colonization of surfaces is a sanitary and industrial issue for many applications, leading to product contamination and human infections. When microorganisms closely interact with a surface, they start to produce an exo-polysaccaridic matrix to adhere to and protect themselves from adverse environmental conditions. This type of structure is called a biofilm. The aim of our work is to investigate novel technologies able to prevent biofilm formation by surface coatings. We coated glass surfaces with melanin-ZnO2, melanin-TiO2, and TiO2 hybrid nanoparticles. The functionalization was performed using cold plasma to activate glass-substrate-coated surfaces, that were characterized by performing water and soybean oil wetting tests. A quantitative characterization of the antibiofilm properties was done using Pseudomonas fluorescens AR 11 as a model organism. Biofilm morphologies were observed using confocal laser scanning microscopy and image analysis techniques were used to obtain quantitative morphological parameters. The results highlight the efficacy of the proposed surface coating to prevent biofilm formation. Melanin-TiO2 proved to be the most efficient among the particles investigated. Our results can be a valuable support for future implementation of the technique proposed here in an extended range of applications that may include further testing on other strains and other support materials.

Two-Phase Extraction Processes, Physicochemical Characteristics, and Autoxidation Inhibition of the Essential Oil Nanoemulsion of Citrus reticulata Blanco (Tangerine) Leaves

Combined ultrasound-microwave techniques and pre-enzymatic treatment (hemicellulase and cellulase) enhance essential oil isolation from Citrus reticulata Blanco (tangerine) leaves (CrBL). Subsequently, synergistic effects of modified amorphous octenyl succinic anhydride starch (OSA-MS), almond oil, and high-energy microfluidics were studied in synergy with ultrasound techniques in the production of CrBL essential oil (CrBL-EO) nanoemulsion (CrBL-EONE). GC-MS was used to study the extraction technique. Dynamic light scattering (DLS) analysis was used with confocal laser scanning microscopy (CLSM) techniques to investigate the nanoemulsion matrices' physical and chemical properties. The D-limonene nanoemulsion (D-LNE) reached the optimal size of droplets (65.3 ± 1.1 r.nm), polydispersity index (PDI) (0.167 ± 0.015), and ζ-potential (-41.0 ± 0.4 mV). Besides, the CrBL-EONE obtained the optimal size of droplets (86.5 ± 0.5 r.nm), PDI (0.182 ± 0.012), and ζ-potential (-40.4 ± 0.8 mV). All the nanoparticle treatments showed significant values in terms of the creaming index (CI%) and inhibition activity (IA%) in the β-carotene/linoleate system with a low degradation rate (DR). The current study's findings showed that integrated ultrasound-microwave techniques and pre-enzymatic treatment could enhance the extraction efficiency of the CrBL-EO. In addition, OSA-MS and almond oil can also be employed to produce CrBL-EONE and D-LNE.

Material gradients in gastropod radulae and their biomechanical significance: a combined approach on the paludomid Lavigeria grandis

The radula, a chitinous membrane spiked with teeth, is the molluscan autapomorphy for the gathering and processing of food. The teeth, as actual interfaces between the organism and the ingesta, act as load transmitting regions and have to withstand high stresses during foraging — without structural failure or high degrees of wear. Mechanisms contributing to this were studied previously in paludomid gastropods from Lake Tanganyika. For some species, gradients in hardness and Young’s modulus along the teeth were detected, enabling the bending and relying of teeth onto the next row, distributing the stresses more equally. The here presented study on one of them — Lavigeria grandis — aims at shedding light on the origin of these functional gradients. The mechanical properties were identified by nanoindentation technique and compared to the elemental composition, determined by elemental dispersive X-ray spectroscopy (EDX, EDS). This was done for the complete radular (mature and immature tooth rows), resulting in overall 236 EDX and 700 nanoindentation measurements. Even though teeth showed regional differences in elemental composition, we could not correlate the mechanical gradients with the elemental proportions. By applying confocal laser scanning microscopy (CLSM), we were finally able to relate the mechanical properties with the degree of tanning. CLSM is a common technique used on arthropod cuticle, but was never applied on radular teeth before. In general, we found that nanoindentation and CLSM techniques complement one another, as for example, CLSM is capable of revealing heterogeneities in material or micro-gradients, which leads to a better understanding of the functionalities of biological materials and structures.

Design of nanostructured coating to prevent biofilm formation on surfaces

Biofilms are surface-attached microbial organizations, where microbial species are enclosed in an exo-polysaccharides matrix. Its removal/prevention from surfaces represents an important challenge and a relevant issue in many fields. In this light, antimicrobial peptides (AMPs) represent good candidates to conventional antibiotics due to their selectivity towards targets (cell membranes), rapid action and low tendency to induce bacterial resistance. In our project we are investigating novel techniques to realise nanostructured coatings on surfaces, in order to prevent bacterial adhesion and biofilm formation. We are focusing our attention on a well-known antimicrobial peptide (Magainin-2). Two bacteria models have been considered: E.coli and P. fluorescens. Our goal is to optimize AMPs grafting on surfaces by means of treatment methodologies based on Oxygen plasma technologies. In this work we present a methodology to investigate anti-biofilm efficiency of the nano-functionalized surfaces, compared with the case of the raw surfaces. Biofilm morphologies can be measured using Confocal Laser Scanning Microscopy and image analysis techniques to quantify its structure by measuring quantitative morphological parameters.

On springtails (Hexapoda: Collembola): a morphofunctional study of the jumping apparatus

BackgroundSpringtails (Hexapoda: Collembola) are tiny organisms that lead a hidden life, mostly occuring deep in the soil and on leaf litter. They have a variety of interesting body morphology patterns, the most famous of which is the catapult-like structure that enables them to jump and flee from predators. This highly specialized jumping apparatus consists of a mobile furca, which when at rest fits into a trigger, "the retinaculum" on the ventral side of the abdomen. Despite the many studies that have attempted to investigate the jumping apparatus, the actual mechanisms involved in the jump, for example the way in which the furca is released by the retinaculum, how and where the mechanisms of spring and hydrostatic pressure originate, are still not properly understood. The morphology of the jumping apparatus of Orchesella cincta was investigated in detail using confocal laser scanning microscopy and MicroCT techniques for 3D reconstruction.ResultsThe morphology of O. cincta with both flexed and extended furca is analysed and described. The abdominal musculature involved in the jumping mechanism and relevant structures of the exoskeleton of retinaculum and furca are described in detail. With the data obtained in this study, hypotheses can be made about (1) where and how the spring and hydrostatic pressure mechanisms originate; (2) which muscles act on the extension and flexion of the furca; (3) which muscles act on the retinaculum and (4) how the retinaculum is released from the furca.ConclusionsThe comparative morphological study proved informative, and shows how springtail jumping involves mechanisms unique to this taxon. Hydrostatic pressure regulation possibly varies between animals with distinct segmentation, and those with fused segmentation. Interesting cuticular characters were revealed, such as basal plates and sclerites related to the construction of the spring mechanism. The present study establishes itself as a model option for future morphofunctional studies on springtail’s jumping. Analysis of videos and images using a high speed camera will be useful for understanding how the jump develops through take-off, aerial and landing phases.

Morphology of the Female Reproductive System of the Soybean Thrips, Neohydatothrips variabilis (Beach, 1896) (Thysanoptera: Thripidae).

Simple SummarySoybean thrips are an important vector of Soybean vein necrosis virus disease, found in all soybean-growing regions of the United States and Canada. The disease reduces the seed oil content and fatty acid profiles in infected plants. It is important to know the morphology of the female reproductive system of soybean thrips to understand the egg-laying mechanism and develop strategies for virus vector management. For this purpose, we used fuchsin staining, paraffin-based histology, dissections, serial block-face scanning electron microscopy and confocal laser scanning microscopy techniques to understand the structure and motorization of the internal and external genitalia of soybean thrips. We also investigated the egg-laying behavior of soybean thrips. The female reproductive system is composed of two ovaries, an oviduct, an accessory gland, an appendage gland, a common oviduct and a vagina. Female soybean thrips lay eggs in the parenchymatous tissues near the veins of the leaves. The appendage gland likely secretes lubrication to facilitate the movement of eggs through the external genitalia. Seven muscles support the movement of eggs from the ovaries to the leaf surface. The anatomy and probable role of each muscle is also described.Soybean thrips (Neohydatothrips variabilis) are an important phytophagous vector of the widely recognized Soybean vein necrosis orthotospovirus (SVNV). Understanding the egg-laying behavior of these thrips could aid in developing strategies for the management of the vector and virus. In this study, we described the egg-laying behavior of N. variabilis and reconstructed the three-dimensional morphology of the female terminalia by using serial block-face scanning electron microscopy (SBFSEM) and confocal laser scanning microscopy (CLSM). The female reproductive system consists of two panoistic ovaries consisting of eight ovarioles. The appendage gland is connected to the ovaries by two muscles, and to the body wall by a single muscle. The spermatheca is connected to the eighth tergum through four branched muscles, to the basivalvulae of the ovipositor by one muscle and to the vagina by a single muscle. The external genitalia are operated by seven muscles. The movement of the eggs inside the ovipositor is achieved by the back and forth “rocking” movement of the first valvulae and valvifer. Eggs are deposited into the parenchymatous tissue alongside leaf veins. To the best of our knowledge, this is the first study describing the internal and external genitalia of N. variabilis.

Surface topography data fusion of additive manufacturing based on confocal and focus variation microscopy.

In this paper, two innovative data fusion methods are proposed for reconstructing the surfaces produced by directed energy deposition (DED) additive manufacturing. The surface topographic data were obtained from confocal laser scanning microscopy (CLSM) and focus variation microscopy (FV). The first method (competitive data fusion) aims to improve the data quality by combining both the advantages of the CLSM and FV techniques, while the second method (cooperative data integration) is designed for generating a single representation that contains not only global information but also local details. The results show that both fusion methods achieved satisfactory results: in the competitive fusion, the fused data preserved the characteristics of FV data while its vertical resolution is also improved by integrating the short waves from the CLSM data; the cooperative data fusion achieved one pixel precision of the surface registration which adopted the feature-based registration method with the help of color image information. The computational complexity is reduced from O((m×n)2) to O(m×n + k). Both proposed data fusion methods provided innovative solutions for the microscopic surface reconstruction and surface representation in multiscales in the field of additive manufacturing.

The contribution mechanism of furfuryl alcohol treatment on the dimensional stability of plantation wood

Furfuryl alcohol polymers (PFA) derived from lignocellulosic biomass have been confirmed as an effective modifier to improve dimensional stability of wood. However, the main reason for the dimensional stability related to this method has remained unclear. In this study, principal consideration of dimensional stabilization was analyzed by a mathematics model regarding anti-swelling efficiency (ASE) and cell wall bulking of furfurylated woods with different weight gain precents (WPGs), and ASE’ obtained by method of Ohmae et al. Simultaneously, confocal laser scanning microscopy and nanoindentation technique were applied to explain the dimensional stability mechanism. Results showed that both ASE and cell wall bulking increased with WPG at 0–69%. A nearly linear (R2 = 0.9690) relationship was observed between ASE and cell wall bulking. ASE’ values at different WPGs were approaching zero. The dimensional stability was dominated by cell wall bulking analyzed through both the mathematical model and small ASE’ values at various WPG. In this study, the mechanistic insights into dimensional stabilization of furfurylated wood could provide highly beneficial for controlling the dimensional stability by cell wall bulking.

Molecular-Level Control over Ionic Conduction and Ionic Current Direction by Designing Macrocycle-Based Ionomers

Poor ionic conductivity of the catalyst-binding, sub-micrometer-thick ionomer layers in energy conversion and storage devices is a huge challenge. However, ionomers are rarely designed keeping in mind the specific issues associated with nanoconfinement. Here, we designed nature-inspired ionomers (calix-2) having hollow, macrocyclic, calix[4]arene-based repeat units with precise, sub-nanometer diameter. In ≤100 nm-thick films, the in-plane proton conductivity of calix-2 was up to 8 times higher than the current benchmark ionomer Nafion at 85% relative humidity (RH), while it was 1–2 orders of magnitude higher than Nafion at 20–25% RH. Confocal laser scanning microscopy and other synthetic techniques allowed us to demonstrate the role of macrocyclic cavities in boosting the proton conductivity. The systematic self-assembly of calix-2 chains into ellipsoids in thin films was evidenced from atomic force microscopy and grazing incidence small-angle X-ray scattering measurements. Moreover, the likelihood of alignment and stacking of macrocyclic units, the presence of one-dimensional water wires across this macrocycle stacks, and thus the formation of long-range proton conduction pathways were suggested by atomistic simulations. We not only did see an unprecedented improvement in thin-film proton conductivity but also saw an improvement in proton conductivity of bulk membranes when calix-2 was added to the Nafion matrices. Nafion–calix-2 composite membranes also took advantage of the asymmetric charge distribution across calix[4]arene repeat units collectively and exhibited voltage-gating behavior. The inclusion of molecular macrocyclic cavities into the ionomer chemical structure can thus emerge as a promising design concept for highly efficient ion-conducting and ion-permselective materials for sustainable energy applications.

Laser gas oxidising of Ti–6Al–4V surface: experimental and numerical investigation

ABSTRACT The laser gas oxidising of Ti–6Al–4V surface was studied. Confocal laser scanning microscopy techniques were used to investigate its two- and three-dimensional surface topographies. Gas flow behaviour was simulated to define the minimum O2 concentration required for Ti–6Al–4V to self-sustain combustion. To understand the flow behaviour of the Ti–6Al–4V fluid, a high-speed camera monitored the evolution of the Ti–6Al–4V fluid, and the flow behaviour was simulated. Experimental and simulation results show that Ti–6Al–4V can self-sustain combustion with a high O2 to Ar gas flow ratio, and the oxidising layer is rough with macroscopic cracks, while a low O2 to Ar gas flow ratio results in a smooth oxidising layer without cracks. Ti–6Al–4V can self-sustain combustion when the O2 concentration exceeds 65%. Furthermore, the laser molten pool and self-sustaining combustion zone mix and flow together, and a large self-sustaining combustion zone leads to a low convection velocity.

A Structural Assessment of Sycamore Maple Bark Disintegration by Nectria cinnabarina

Previous phytopathological studies of the fungal pathogen Nectria cinnabarina have been focused on its distribution and host diversity but little is known about the spread of this pathogen and the defence responses of forest trees to an infection inside host tissues. Histopathological alterations of bark, periderm, phloem and woody tissues were investigated in sycamore maple (Acer pseudoplatanus) branches following their natural attack by the advanced anamorph and teleomorph developmental stages of the fungus. Light, fluorescence, confocal laser scanning and scanning electron microscopy techniques supplemented by X-ray micro-computed tomography imaging were used to distinguish between healthy and disintegrated plant tissues. The intercellular spread of fungal hyphae was found primarily in the phelloderm. Expanding hyphae aggregations produced ruptures in the phellem and the disintegration of both phellogen and phellodermal parenchyma cells in close proximity to the expanding fruiting bodies of the fungus. Thicker hyphae of the teleomorph fungal stage heavily disintegrated the phelloderm tissues and also induced enhanced sclerification of the nearby phloem tissues that limited the spread of the infection into the sieve tubes. Both the intercellular and intracellular spread of hyphae inside the peripheral parts of sclereid clusters led to the disintegration of the compound middle lamellae but the hyphae were only rarely able to pass through these structural phloem barriers. The massive fungal colonization of both lumens and disintegrated tangential cell walls of ray parenchyma cells resulted in severe cambial necroses. Although the hyphae penetrated into the outermost annual growth rings of the xylem, no cell wall disintegration of the parenchyma cells, vessels and fibres was revealed. Despite the local cambial necroses and severe phloem ray disintegration, the bark remained attached to the examined branches and no bark cankers were formed.

An efficient and stable magnetic nano-biocatalyst for biodiesel synthesis in recyclable ionic liquids

The process of jatropha oil transesterification was studied for biodiesel production. Rhizomucor miehei lipase (RML) immobilized on 3-aminopropyltriethoxysilane functionalized magnetic Fe3O4 was employed as a biocatalyst. The immobilized lipase was confirmed by scanning electron microcopy (SEM), Fourier transform infrared spectra (FT-IR), thermogravimetric analysis (TGA), vibrating sample magnetometer (VSM), and confocal laser scanning microscopy (CLSM) techniques. The efficient biodiesel synthesis in ionic liquids using immobilized RML was demonstrated. Fifteen kinds of ionic liquids based on different alkyl chain lengths of the methyl imidazolium cation ([C2MIM], [C4MIM], [C8MIM], [C12MIM], and [C16MIM]) combined with [NTf2], [N(CN)2], [PF6] or [BF4] anions were assayed as reaction media for RML producing biodiesel. The highest synthetic activity of immobilized RML appeared in [BMIM][PF6], which is more than five times higher than the free enzyme. The enzyme catalytic activity remained 60% even after the magnetic nano-biocatalyst and ionic liquid had been reused 5 times in a 48-h reaction cycle. In addition, the immobilized enzyme exhibited excellent storage stability with almost no decrease in catalytic activity after storing at − 20 °C for 98 days.

The fundamental difference between cleavage and hydrogen-assisted quasi-cleavage in ferritic materials revealed by multiscale quantitative fractographic and side surface characterization

The fracture surfaces and cracks in technically pure iron, Fe-2.5% Si alloy and low-carbon steel grade S235JR produced by quasi-cleavage (QC) hydrogen-assisted cracking (HAC) as well as by true cleavage (TC) were studied and compared on different scales through the quantitative and qualitative fractographic and side surface examinations. The mixed type QC/TC fracture surfaces were obtained using two-step mechanical testing methods involving the tension of the through-notched specimens under in-situ hydrogen charging followed by rapid fracture in liquid nitrogen. The side surface and fractographic examinations, including the quantitative facets analysis, were conducted using scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and electron-backscattering diffraction (EBSD) technique. It is established, that the QC and TC fracture surfaces demonstrate fundamentally different morphological and topological features in all materials investigated. It is found that, depending on the material, the average angle of misorientation (AM) between the QC facets is 1.5–2 times lower than that of the TC facets. Thus, it is concluded that the crystallographic orientation of QC facets is generally different from TC. Owing to this feature, the path of the QC HAC through the microstructure of all investigated materials is featured by the remarkably lower spatial out-of-plane deviation on the scale equal to or larger than a few facets. Conversely, on the scale of an individual facet, the path of QC HAC deviates to a greater extent than that of the TC crack. This fundamental difference between QC HAC and TC cracking is found to be not significantly influenced by the chemical composition of the ferritic alloys.

Quadruple-action coatings provided by doping epoxy with inhibitor laden clay nanotubes functionalized with layer-by-layer of cross-bridged chitosan and anionic polyelectrolytes

The present study divulges the quadruple-protective-action including combined enhanced barrier, active feedback protection, self-repairing and anti-fouling properties of epoxy composite coatings reinforced with benzotriazole-laden clay nanotubes capped with ferric ion and wrapped with layer-by-layer (LBL) assembly of chitosan/polyacrylic acid (CTS-PAA-CTS-PAA) and chitosan/polymethacrylic acid (CTS-PMMA-CTS-PMMA) polyelectrolytes cross-bridged with (+D)-epichlorohydrin. UV–vis spectrophotometric technique reveals the pH-responsive release of the loaded benzotriazole, while scanning vibrating electrode technique (SVET), electrochemical impedance spectroscopy (EIS) and optical techniques reveal the combined active feedback corrosion suppression and self-repairing of impressed defects. Additionally, confocal laser scanning microscopy (CLSM) and SEM techniques reveal the anti-bacterial properties. The experimental results confirmed that the new composite coatings exhibited unprecedented quadruple-action protection.