Multiphoton microscopy (MPM) is a cutting-edge nonlinear microscopy technique known for its exceptional capabilities in imaging dense tissues, making it an ideal tool for exploring human kidney tissue. This study delves into the application of MPM in evaluating renal cell tumors and characterizing tumor capsules. MPM encompasses two core methods: two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG). It leverages femtosecond lasers to provide deep tissue imaging while minimizing phototoxicity, making it well-suited for live tissue analysis. The study analyzed 100 formalin-fixed paraffin-embedded (FFPE) kidney tissue samples, ensuring compliance with ethical standards. Using a Ti:Sapphire laser for femtosecond pulses and a variety of optical components and filters, the researchers captured TPEF and SHG signals, facilitating image analysis. Normal kidney tissue exhibited distinctive structures like glomeruli and tubules under MPM. Renal cell carcinoma (ccRCC) displayed unique TPEF patterns with eccentrically located nuclei and indistinct cytoplasmic membranes. Papillary RCC (pRCC) exhibited a papillary pattern with dark nuclei, while chromophobe RCC (chRCC) showcased distinct features like thick cell membranes, strong TPEF signal in nuclei, and granular cytoplasm. Renal oncocytoma (RO) presented strong TPEF signals in cuboidal cells with microcyst-like arrangements. Overall, MPM proved invaluable in assessing human kidney tissue, offering distinct TPEF patterns for different renal cell tumor types. This non-invasive technique has the potential to enhance pathological examinations and advance our understanding of complex biological processes, ultimately contributing to improved diagnostics and treatment strategies for renal diseases.

Background. Escalating global plastic production, expected to reach 34,000 million tons by 2050, poses a significant threat to human and environmental well-being, particularly in aquatic ecosystems. Microplastics (MP) and nanoplastics (NP), which originate from the degradation of plastics, are of concern due to their potential bioaccumulation and uptake of pollutants. This study addresses identification methods and focuses on insect meal, a raw material for aquaculture feed. Methods. By using different techniques, the study was able to detect MP and NP in insect meal samples. Chemical digestion with KOH at 60°C efficiently removed organic matter without affecting the synthetic polymer polyethylene (PE). Filtration, confocal Raman microscopy, SEM and TEM were used for comprehensive analysis and integrity tests on PE films were performed using Raman and FTIR spectroscopy. The results showed the presence of PE microplastic particles in the insect meal, which was confirmed by correlative Raman and SEM mapping on a positively charged surface. In addition, the increased resolution of the Raman microscope identified submicrometric PE NP (800 nm). Transmission electron microscopy (TEM) with energy dispersive X-ray spectroscopy (EDX) confirmed plastic-like structures in the insect meal, highlighting the presence of PE plastics characterized by irregular shapes and some agglomeration. The higher carbon concentration in the EDX analysis supported the plastic nature, which was also confirmed by Raman spectroscopy. Conclusions. The study provides a robust method for the detection of MP and NP in insect meal and provides valuable insight into the possible presence of plastics in insect-based aquafeeds. The combination of different analytical methods increases the reliability of the results and sets the stage for future investigations that could focus on the quantification of NP and the assessment of their potential environmental impact.

This article explores the transformative role of neural networks in the realm of biology, particularly within microscopy image processing and illustrations. Neural networks have revolutionized cell segmentation and analysis, enabling precise delineation of cell boundaries and tracking of cellular behaviours. They excel in detecting subcellular structures, unravelling intricate organelle interactions. In 3D image reconstruction, neural networks navigate volumetric datasets, enhancing our understanding of spatial cellular architecture. These networks enhance disease diagnosis by identifying anomalies and irregularities, potentially revolutionizing early detection and classification. Moreover, neural networks restore and enhance microscopy images, unveiling hidden details. Lastly, they bridge art and science, fostering captivating biological art and enriching science communication. As neural networks evolve, they promise a future of limitless possibilities, weaving together technology, science, and art to illuminate the microscopic realm in unprecedented ways.

The natural flavonoid epigallocatechin gallate (EGCG) has a wide range of biological activities including binding to nucleic acids; however, mechanisms of interaction of EGCG with DNA organized in chromatin have not been systematically studied. In this work, interaction of EGCG with chromatin in cells, and with nucleosomes and chromatosomes in vitro was studied using fluorescent microscopy and single particle Förster resonance energy transfer approaches, respectively. EGCG effectively penetrates into the nuclei of living cells and binds to DNA there. Interaction of EGCG with nucleosomes in vitro induces large-scale, reversible uncoiling of nucleosomal DNA that occurs without dissociation of DNA or core histones at sub- and low-micromolar concentrations of EGCG. EGCG does not reduce the catalytic activity of poly(ADP-ribose) polymerase 1 (PARP1), but causes modulation of the structure of the PARP1-nucleosome complex. EGCG significantly changes the structure of chromatosomes, but does not cause dissociation of the linker histone. Reorganization of nucleosomes and chromatosomes by EGCG might facilitate access of protein factors involved in DNA repair, replication and transcription to DNA and thus might contribute to the modulation of gene expression by EGCG that was reported earlier.

Neat polyimide films are known to be dense and rigid. They are therefore not suitable for use in membranes, sensors and sustainable energy storage applications. In this study a novel technique has been used to simulta-neously improve the rigidity, damping ability and impact resistance of polyimide membranes. It is demonstrated that dispersion of a small amount of polyaniline copolymer-modified clay of about 0.25-0.5 wt.% into the pol-yimide matrix, resulted in enhanced storage modulus while maintaining high damping ability and glass transition temperature, Tg. Novel polyimide/substituted polyaniline-copolymer-clay nanocomposite membranes containing poly(N-ethyl-aniline-co-aniline-2-sulfonic-acid-modified-clay (SPNEAC) was successfully prepared and in-corporated into polyimide matrix to form modified clay/polyimide nanocomposites. UV-Vis analysis of the nanocomposite films show that the optical transparency of the SPNEAC-PI nanocomposite membranes de-creased with increasing SPNEAC concentration due to the high UV-Vis absorption of SPNEAC. Transmittance of about 3 % was observed in the nanocomposite membrane containing 5wt.% modified clay at 500 nm wave-length, which is significantly lower than that for the neat PI membrane of about 36 %. The dispersion of SPNEAC containing high concentration of clay (≥ 40 wt.% clay), SPNEAC2 in polyimide matrix, resulted in attainment of a higher degree of imidization than was possible for the neat organoclay/polyimide nanocomposite. This behavior is believed to be due to the synergistic interaction between PI and polyaniline copolymer modified clay. The viscoelastic property of polyimide and the nanocomposite membranes was measured by using the dynamic mechanical spectroscopy, DMS. It was shown that the glass transition temperature, Tg of the nano-composites decreased, while the damping ability and impact energy increased with increasing weight fraction of fillers.

Supramolecular solvent (SUPRAS) has received increasing interest as an innovative, efficient, and green solvent for the effective extraction and separation of bioactive compounds from natural resources. However, studies on the use of SUPRAS for the extraction of phenolic components from plants, which are highly valued in food products due to their excellent antioxidant capacity, are still lacking. The present study developed a green ultrasound-assisted and efficient SU-PRAS method to simultaneously determine three phenolic acids in Prunella vulgaris by high-performance liquid chromatography (HPLC) and the experimental parameters were optimized in detail. The efficiency and antioxidant properties of the phenolic compounds obtained by different extraction methods were also compared. Under the optimal conditions, the extraction efficiency of SUPRAS, prepared with octanoic acid reverse micelles dispersed in ethanol-water, was substantially higher than conventional organic reagents. Moreover, this SUPRAS method also showed more significant antioxidant capacities. The confocal laser scanning microscopy (CLSM) images presented the spherical droplet structure of SUPRAS (A well-defined circle), which coincided with the position of the fluorescence position of phenolic acids. The phenolic acids were encapsulated in the vesicles by the SUPRAS droplets, which indicated the efficient extraction capacities of SUPRAS for the target compounds. Finally, combined with the CLSM method, the mechanism of the proposed method was further clarified, and the superior extraction performance of SUPRAS was theoretically demonstrated through molecular dynamics simulations. In comparison with conventional methods, the higher extraction efficiency of SUPRAS can be illustrated by the larger solvent contact surface area, more types of hydrogen bonds between extractants and supramolecular solvents, and stronger and more stable interaction forces. The results of the theoretical studies are in rational agreement with the experimental outcome.

The quality assurance of malaria microscopy unit was established by National Malaria Control Program, Ministry of Health (MoH) in 2009 to respond to the internal malaria microscopy validation process, in microscopy training for malaria diagnosis in the country, monitoring and supervision, and other relevant activities regarding laboratory diagnosis of malaria in Timor-Leste is described. Trained laboratory technicians who are working in the relevant unit. There are two ways of malaria microscopy validation process such as onsite validation and monthly validation after received malaria blood smear from senior analyst at the municipality level. The aim of this work is to describe the quality assurance of malaria microscopy system in Timor-Leste. The management of quality assurance service support, the importance of quality assurance of malaria microscopy, the role of quality assurance of malaria microscopy, challenge and opportunities encountered by quality assurance microscopy unit, National Health Laboratory are highlighted. Key words: Quality assurance of malaria microscopy, system, Timor-Leste.

In the present research, groups of nanolayered structures and nanohybrids based on organic green dyes and inorganic species are designated to act as fillers for PVA for inducing new optical sites and increasing its thermal stability through producing polymeric nanocomposites. In this trend, different percentages of Naphthol green B were intercalated as pillars inside the Zn-Al nanolayered structures to form green organic-inorganic nanohybrids. The two-dimensional green nanohybrids were identified by X-ray diffraction, TEM and SEM. According to the thermal analyses, the nanohybrid, which has the highest amount of green dyes, was used for modifying the PVA through two series. In the first series, three nanocomposites were prepared depending on the green nanohybrid as prepared. In the second series, the yellow nanohybrid, which was produced from the green nanohybrid by thermal treatment, was used to produce another three nanocomposites. The optical properties revealed that the polymeric nanocomposites depending on green nanohybrids became optical-active in UV and visible regions because the energy band gap decreased to be 2.2 eV. In addition, the energy band gap of the nanocomposites depended on yellow nanohybrids was 2.5 eV. The thermal analyses indicated that the polymeric nanocomposites are thermally more stable than that of the original PVA. Finally, the dual functionality of organic-inorganic nanohybrids that produced from the confinement of organic dyes and the thermal stability of inorganic species converted the non-optical PVA to optical-active polymer in a wide range with high thermal stability

Raman spectroscopy (RS) turns out to be a promising tool for cancer di- agnosis. In particular, in the last years several studies have demonstrated how the diagnostic performances of Raman Spectroscopy can be significantly improved by employing Machine Learning (ML) algorithms for the interpre- tation of Raman-based data. In line with these findings, in this paper, we demonstrated how RS coupled with ML allowed to accurately distinguish the three grades of Chondrosarcoma and to distinguish Chondrosarcoma and a benign counterpart called Enchondroma. We obtained such results by ana- lyzing a dataset composed of a relatively small number of Raman spectra, collected in a previous study. Such spectra were acquired from micromet- ric tissue sections with a Confocal Raman Microscope. In particular, we tested the classification performances of a Support Vector Machine and a Random Forest Classifier, as representative of Machine Learning, and two versions of the Multi-Layer Perceptron, as representative of Deep Learning (DL). These models showed excellent classification performances, especially those belonging to DL, with accuracy reaching 99.7%. This outcome makes the aforementioned models a promising route for future improvements of di- agnostic devices focused on detecting bone cancerous tissues. In addition, we highlighted how the ML models studied resulted in slightly worse classifi- cation performances in comparison to DL. Alongside the diagnostic purpose, the aforementioned approach allowed to identify characteristic molecules, i. e. amino acids, nucleic acids and bioapatites, relevant for the obtainment of the final diagnostic response, through the analysis of the so-called Per- mutation Feature Importance. Permutation Feature Importance could hence represent a promising parameter for the understanding of the biochemical processes on the basis of the tumor progression. In turn, the spectral bands highlighted by Permutation Feature Importance could represent precious in- dicators in the attempt to restrict the spectral acquisition to specific Raman bands. This last objective could help to reduce the amount of experimental data needed to obtain an accurate final grading outcome, with consequent reduction of the computational cost.

Three-layer structures based on various multi-component films of III–V semiconductors heavily doped with Fe were grown by pulsed laser sputtering of InSb, GaSb, InAs, GaAs and Fe solid targets. The structures comprising InAsSb:Fe, InGaSb:Fe and InSb:Fe layers with Fe concentrations up to 24 at. % separated by GaAs spacers were deposited on (001) i-GaAs substrates at 200ºC. Transmission electron microscopy showed that the structures have a rather high crystalline quality and do not contain secondary-phase inclusions. X-ray photoelectron spectroscopy investigations revealed significant diffusion of Ga atoms from the GaAs regions into the InAsSb:Fe layers, which has led to the formation of an InGaAsSb:Fe compound with a Ga content up to 20 at. %. It has been found that the ferromagnetic properties of the InAsSb:Fe magnetic semiconductor improve with increasing Sb:As ratio. It has been concluded that the indirect ferromagnetic exchange interaction between Fe atoms occurs predominantly via Sb atoms.

This work deals with the analysis of phase composition of sintered Ti-Al-C composite materials under high voltage electrical discharge. The samples of Ti-Al-C powder composites were tested by SEM, Raman spectroscopy and XRD. It was determined that the treatment of the powder by high voltage electrical discharge (HVED) and further sintering at high temperatures using the method of spark plasma sintering (SPS) encourages the formation of the intermetallic reinforcing phases, carbides and different nanocarbon structures like graphene and fullerenes, as well as pure graphite. Intermetallic phases and nanocarbon structures improve mechanical and physical properties of the composites. By using experimental methods mentioned above, the phase composition of Ti-Al-C powder composites obtained at different sintering temperatures was determined.

Tellurium exhibits exceptional intrinsic electronic properties. However, investigations into the modulation of tellurium's electronic properties through physical modification are notably scarce. Here, we present a comprehensive study focused on the evolution of the electronic properties of tellurium crystal flakes under plasma irradiation treatment by employing conductive atomic force microscopy and Raman spectroscopy. The plasma-treated tellurium experienced a process of defect generation through lattice broken. Prior to the degradation of electronic transport performance due to plasma irradiation treatment, a remarkable observation emerged: in the low-energy region of hydrogen plasma-treated tellurium, a notable enhancement in conductivity was unexpectedly detected. The mechanism underlying this enhancement in electronic transport performance was thoroughly elucidated by comparing it with the electronic structure induced by argon plasma irradiation. This study not only fundamentally uncovers the effects of plasma irradiation on tellurium crystal flakes, but also unearths an unprecedented trend of enhanced electronic transport performance at low irradiation energies when utilizing hydrogen plasma. This abnormal trend bears significant implications for guiding the prospective application of tellurium-based 2D materials in the realm of electronic devices.

The mitochondrial network (MN) is a dynamic structure undergoing constant remodeling in the cell. It is assumed that the violation of MN may be associated with various pathologies, including Parkinson’s disease (PD). Using automatic image analysis and super-resolution microscopy, we have assessed the MN parameters in fibroblasts from patients with established hereditary PD mutations (associated with PINK1, LLRK2, α-synuclein, PINK1 and Parkin simultaneously proteins) under normal conditions and after hydrogen peroxide-induced stress. Fibroblasts with Pink1/Parkin mutation are the most different in morphology from fibroblasts obtained from conditionally healthy donors: MN is larger, contains longer mitochondria and accumulated individual mitochondria. In addition to MN, we evaluated other cellular parameters, such as: cytosolic and mitochondrial ROS production and mitochondrial membrane potential. It has been shown that mitochondria of fibroblasts with mutations in genes encoding PINK1, α-synuclein and Pink/Parkin tends to hyperpolarization and cytosolic ROS overproduction, while mitochondrial ROS production was higher only in fibroblasts with PINK1 and α-synuclein mutation.

Smelting used to be less efficient in the past; therefore, wastes obtained from historical processing at smelter plants usually contain certain quantities of valuable metals. Upon the extraction of useful metal elements, metallurgical slag can be repurposed as an alternative mineral raw material in the building sector. A case study was conducted, which included an investigation of the physico-chemical, mineralogical, and microstructural properties of Pb-Zn slag found at the historic landfill near the Topilnica Veles smelter in North Macedonia. The slag was sampled using drill holes. The mineralogical and microstructural analysis revealed that the Pb-Zn slag is a very complex and inhomogeneous alternative raw material with utilizable levels of metals, specifically Pb (2.3 wt%), Zn (7.1 wt%), and Ag (27.5 ppm). Crystalline mineral phases wurtzite, sphalerite, galena, cerussite, akermanite, wüstite, monticellite, franklinite, and zincite were identified in the analyzed samples. The slag’s matrix consisted of alumino-silicates, amorphous silicates, and mixtures of spinel and silicates. Because the extraction of Pb, Zn, and Ag has economic potential, the first reutilization stage will be to convert metal concentrates into their collective concentrate, from which the maximum amount of these valuable elements may be taken out. This will include combination of gravity concentration and separation techniques. The next step is to assess the Pb-Zn slag’s potential applications in civil engineering based on its mineralogical and physico-mechanical properties. Alumino-silicates, amorphous silicates, and mixtures of spinel and silicates present in Pb-Zn slag, which contain high concentrations of SiO2, Al2O3, CaO, and Fe2O3, are suitable for use in cementitious building composites. The goal of this research is to suggest a solution for closing the circle of slag's reutilization in terms of Zero Waste principles, so it is critical to thoroughly investigate the material and establish methods and preparation processes, as well as ways of concentrating useful components into commercial products.

Background: MRI-guided prostate biopsies from visible tumor-specific lesions (TBx) can be used to diagnose clinically significant carcinomas (csPCa) requiring treatment more selectively than conventional systematic biopsies (SBx). Ex vivo fluorescence confocal microscopy (FCM) is a novel technique that can be used to examine TBx prior to conventional histologic workup. Methods: TBx from 150 patients were examined with FCM on the day of collection. Preliminary findings were reported within 2 hours of collection. The results were statistically compared with the final histology. Results: 27/40 (68%) of the csPCa were already recognized in the intraday FCM in accordance with the results of conventional histology. Even non-significant carcinomas (cisPCa) of the intermediate and high-risk groups (PSA > 10 or 20 ng/ml) according to conventional risk stratifications were reliably detectable. In contrast, small foci of cisPCa were often not detected or were difficult to distinguish from reactive changes. Conclusion: The rapid reporting of preliminary FCM findings helps reduce the psychological stress on patients an can improve the clinical management of csPCa. Additional SBx can be avoided in individual cases, leading to lower rates of complications and scaring in the future surgical area. Additional staging examinations can be arranged without losing time. FCM represents a promising basis for future AI-based diagnostic algorithms.

Both the use of plastic and its decomposition products lead to the distribution of plastic all over the earth and finally to the uptake by all kind of living beings including humans. Still, it is widely unknown what risks harbor the widespread uptake of plastics for human health, espe-cially regarding contributing factors like size, shape and surface composition. We assessed the uptake of polymethylmethacrylate (PMMA)-nano- and microbeads for HEK293-, A549- and MRC5 cells. Via confocal microscopy, we localized multiple PMMA-beads inside the cytosol of cells. Uptake of PMMA-beads did not alter cell growth and cell division, implying no short-term toxicity towards human cells. Further, we used a cAMP response element binding protein (CREB)-mediated reporter assay to assess whether internalized PMMA-nanobeads alter cell-signaling pathways. In contrast to the in vitro transcription, where the addition of PMMA-nanobeads abolished the transcription, no changes regarding CREB-mediated cell sig-naling are given in HEK293-cells. Our data led to the assumption that PMMA-nano-and mi-crobeads are internalized via endocytosis and end up as lysosomes within the cells cytosol. Therefore, we concluded differences regarding the surface composition of the PMMA-nanobeads mainly affect its potential to alter cell signaling. These findings emphasize the key role the sur-face composition plays regarding microplastic and its risks for human health.

Epitaxial lithium ferrite (LiFe5O8) films with different thicknesses have been successfully fabricated on SrTiO3 (001) substrates by magnetron sputtering deposition technique. The microstructural and magnetic properties are characterized by advanced transmission electron microscope and magnetic measurement device. It was found that the formation of structural defects can be influenced by the thickness of the film. Apart from the misfit dislocations, the orientation domains form in thinner film and twin boundaries appear in thicker film, respectively, contributing to the misfit strain relaxation in the heterosystem. The magnetic measurement shows that the thinner films have enhanced magnetization and a relatively lower coercive field compared with the thicker films containing the antiferromagnetic twin boundaries. Our results provide a way for tuning the microstructure and magnetic properties of lithium ferrite films by changing the film thickness.

Engineering application of additively manufactured (AM) metallic materials is quite limited by their fatigue behaviors which are very inconsistent with that of conventionally wrought or cast ones. Here, based on advanced material characterization techniques, such as focused ion beam (FIB), scanning electron microscopy (SEM) and transmission electron microscopy (TEM), the mi-crostructures underneath fracture surfaces were thoroughly investigated in an AM aluminium (AlSi10Mg) alloy with horizontal and vertical building orientation endured very-high-cycle fa-tigue (VHCF) loading under stress ratios R = –1, 0 and 0.5. Two VHCF failure specimens A and B were representatively selected to further SEM examine and TEM samples preparation via FIB milling. Specimen A was horizontally printed and failed at R = –1; specimen B was vertically printed and failed at R = 0. TEM samples A1 and B1 were lifted from locations near the crack ini-tiation sites on the fracture surfaces of specimens A and B; The locations of TEM samples A2 and B2 kept away from the crack origin sites but still within the “fish-eye” region of crack steady growth. TEM observations show that there was no characteristic microstructure induced by VHCF in different oriented specimens and under various R values.

The aim of the paper is to analyze the experimental results of the influence of elevated temperatures and strain rate on the mechanical and structural properties of steel 42CrMo4. The experiments were based on uniaxial tension and compression tests at high temperatures between 700 °C and 1000 °C and strain rates range 0.0018 - 0.1 s^-1. The influence of temperature and strain rate on yield stress, strain to fracture, hardness, structural changes, and fracture characteristics were analyzed. Due to the dynamic recrystallization phenomenon present during the hot tensile tests, an increase in the flow stress is observed at the beginning of the deformation, after which it decreases until the fracture. By increasing the deformation temperature from 700 °C to 1000 °C, the tensile stress decreases significantly for all strain rates. The increase in the strain rate leads to the increase in the tensile stress. In compression tests by increasing the strain rate, the true strain is slightly increasing, but this depends on the temperature. The non-uniformity of deformations obtained at different values of the strain rate and temperature were also analyzed. Analysis by scanning electron microscopy showed the ductile behavior of the material. The degree of damage of the material caused by the presence of cavities increases by increasing the deformation temperature. For all the presented deformation conditions, the presence of the fracture through the ductile fracture mechanism was produced by the localized of necking and the coalescence of microvoids. By increasing the deformation temperature and reducing the strain rate, the fracture behavior of 42CrMo4 steel can be improved.

The identification of a new virus Kiparis-144 (KiparisV) in a half-fed tick I persulcatus, removed from a person in the south of the Russian Far East, determined the goal of further research - to study its role in human pathology and the characteristics of its behavior in the tick microbiome. An ELISA study of 439 blood samples from people affected by tick bites indicated a fairly frequent (up to 16.4±1.7%) encounter of people with the KiparisV virus in the territory of natural foci of the Primorsky Territory. It has been shown that this virus, when mono-infected, can cause clinical symptoms characteristic of acute respiratory diseases. During co-infection under the influence of the KiparisV, there was an increase in the activity of Borrelia spp. with the constant production of antibodies to them only of the IgM class and clinical symptoms of Lyme disease for one year (observation period). Electron microscopy of the KiparisV found in a tick removed from a patient showed a picture of the relationship of the KiparisV (up to 100 nm in size) with unknown representatives of the tick microbiome - pathogens of a viral or bacterial nature. In a comparative aspect, a similar electron microscopic picture of the interaction of the KiparisV virus with a known pathogen of viral nature, the ectromelia virus (ECTV), is presented. The importance of the electron microscopy method in describing the tick microbiome is shown. Thus, it has been established that when co-cultivated in the system the helper-virus KiparisV probably tends to use another pathogen of the tick microbiome for its reproduction, influencing its pathogenicity.

Transmission electron microscopy methods were used to analyze the cementite substructure in the head of special-purpose long rails of DT400IK category made of hypereutectoid steel after long-term operation on an experimental track on the Russian Railways ring (the passed tonnage is 187 million tonnes). It is noted that the study of various aspects of cementite – its structure, morphology, chemical composition, crystal lattice defects is relevant. The steel structure is represented by three morphological components at a distance of 10 mm from the sample surface: lamellar perlite, fractured and fragmented perlite. The volume fraction of lamellar perlite in the material is 65%. It is shown that after operation, the cementite plates are bent and separated by ferrite bridges. In the plates of ferrite and cementite, a dislocation substructure is formed which is of chaotically distributed and network type in ferrite and of ordered type in cementite. An increased density of dislocations at ferrite-cementite interfaces compared to the volume of ferrite plates was noted. Two possible mechanisms of deformation transformation of lamellar perlite grains are indicated: fracture of cementite plates and carbon pulling out from the lattice of the carbide phase. It is indicated that in the dissolution of cementite plates, the interfacial boundaries “α-phase-cementite” play an important role. The removal of carbon from cementite plates occurs most intensively near defects in ferrite and cementite. The formed nanosized particles of tertiary cementite are unevenly distributed in the ferrite plates, most of them are observed at the locations of ferrite subgrains and interfacial boundaries. This results in non-uniform diffraction contrast in dark-field images of cementite plates. Nanosized particles of cementite can be taken out into the interlamellar space of pearlite colonies in the process of dislocation slip, or they are formed as a result of deformation decomposition, which is less likely. Fragmentation of ferrite and cementite plates is revealed and azimuthal components of total misorientation angles are estimated. The mechanisms of mass transfer of carbon atoms over interstitial sites, deformation vacancies, dislocation tubes, grain boundaries and fragments are considered. According to all the established patterns of the cementite substructure transformation, a comparison with the results for rails made of hypoeutectoid steel was performed.

Changing the film thickness to manipulate the microstructural properties has been considered as a potential method in practical application. Here, we report that atomic-scale structural properties regulated by film thickness in NiCO2O4(NCO)/CuFe2O4(CFO) bilayer heterostructure prepared on (001)-MgAl2O4 (MAO) substrate by means of aberration-corrected scanning transmission electron microscopy (STEM). The misfit dislocations at NCO/CFO interface and antiphase boundaries (APBs) bound to dislocations within the films both are found in NCO (40 nm)/CFO (40 nm)/MAO heterostructure to contribute to relaxation of mismatch lattice strain. In addition, the non-overlapping a/4[101]-APB is found and the structural transformation of this kind of APB is resolved at atomic scale. In contrast, only the interfacial dislocations form at interface without the formation of APBs within the films in NCO (10 nm)/CFO (40 nm)/MAO heterostructure. Our results provide evidence that formation of microstructural defects can be regulated by changing film thickness to tune magnetic properties of epitaxial bilayer spinel oxide films.

Purpose: To evaluate the relationship between corneal nerve structure and function in a veteran population. Methods: 83 veterans (mean age: 55 ± 5 years) seen at the Miami Veterans Affairs (VA) eye clinic were included in the study. Each individual filled out questionnaires to evaluate ocular symptoms (5-Item Dry Eye Questionnaire, DEQ5, Ocular Surface Disease Index, OSDI) and ocular pain (Numerical Rating Scale, NRS, Neuropathic Pain Symptom Inventory modified for the Eye, NPSI-Eye). Individuals also underwent an ocular surface examination that captured functional nerve tests including corneal sensation, corneal staining, and Schirmer test for tear production. Corneal sub-basal nerve analysis was conducted using in vivo confocal microscopy (IVCM) images with corneal nerve density, length, area, width, and fractal dimension captured. IVCM and functional corneal metrics from the right eye were examined using correlational and linear regression analysis. Results: Most corneal structural metrics were not related to functional metrics, except for weak correlations between various IVCM metrics and tear production. In addition, corneal nerve fiber area was positively related to corneal sensation (r = 0.3, p = 0.01). On linear regression analyses, only corneal fractal dimension remained significantly related to tear production (β = -0.26, p = 0.02) and only corneal nerve fiber area remained significantly related to corneal sensation (β = 0.3, p = 0.01). Conclusions: Most corneal nerve structural metrics did not relate to functional metrics in our veteran population, apart from a few weak correlations between structural metrics and tear production. This highlights caution in using corneal nerve anatomy to predict corneal function.

This study aims at relating the microstructural arrangement, in particular the symmetry materialised by filament sequencing in fused filament fabrication process on mechanical behaviour of printed polyamide. Dogbone structures are printed using various printing temperatures, which are combined with part orientation and raster angle that represent the in-plane and out-of-plane symmetrical arrangement of filament. Mechanical testing is conducted on both as-received filaments and printed structures to derive the effect of filament arrangement symmetry and process-generated defects on mechanical loss. In addition, microstructural analysis using scanning electron microscopy is used to share more light on the filament arrangement and their consequence on deformation mechanisms with respect to the printing conditions. The results show that 3D printed polyamide-based material exhibit remarkable tensile performance with strain stiffening behaviour and large elongation at break thanks to particular filament layout. Among the considered printing conditions, the part orientation is found to have the largest influence on tensile behaviour, which modulates the behaviour from complete restoration of the filament performance to mechanical loss.

In this work, we have studied microstructure and unusual ferromagnetic behavior in cobalt (Co)-implanted epitaxial tin dioxide (SnO2) films. The study was aimed to provide a comprehensive understanding of the interplay between cobalt implant distribution, crystal defects, such as oxygen vacancies, and magnetic properties in Co-implanted SnO2 films, which has potential applications in spintronics and memristor devices. The scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), vibrating sample magnetometry (VSM), differential thermo-magnetic analysis (DTMA), and ferromagnetic resonance (FMR) were employed for investigations of SnO2 films implanted with 40 keV Co+ ions at room or the elevated temperatures of substrate. The experimental data showed that the Co ion implantation with high fluence of 1.0 1017 ions/cm2 induces significant changes in the microstructure of SnO2 films, leading to the appearance of ferromagnetism with the Curie temperature significantly above room temperature. Strong influence of implantation temperature and subsequent high-temperature annealing in air or in vacuum on magnetic properties of Co-implanted SnO2 films were established. Moreover, the chemical effect of ethanol on FMR spectra were observed. The obtained results are discussed within the model of two magnetic layers with different concentration and valence states of the implanted cobalt and with high content of oxygen vacancies.

Glycerol represents a useful functional component of heat-transfer fluids. Glycerol is known to be triboelectrically active. We report how the flow of glycerol, stopped in a cylindrical coiled pipe, influences horseradish peroxidase (HRP) enzyme. Namely, glycerol flow was pumped through the coil and then stopped. After that, we incubated the 0.1 µM solution of HRP near the side of the coil, and then adsorbed HRP onto mica. This operation markedly increased the HRP aggregation on mica in comparison with the control sample. We explain this phenomenon by the influence of triboelectrically induced electromagnetic fields. Our results can be useful in the development of flow-based systems for biosensors and bioreactors.

The as-received commercial Inconel 718 material was solid solution heat treated (ST), cold rolled (CR), and precipitation hardened (PH) to investigate the effects of deformation on tensile properties, hardness, and texture. Three sets of specimens (0%, 20% and 50% CR) were STed at 1100 C/1h, CR and aged (720 °C/8 h + 650 °C/8 h) for the analysis. Ultimate tensile strength (UTS), 0.2% yield strength (YS) and elongation of 50% deformed condition were 1645 MPa, 1512 MPa, and 3.8 %, respectively. The 20 % deformation resulted in a balanced UTS (1348 MPa), YS (1202 MPa) and elongation (11%). Contributions of precipitation hardening to the strength was decreasing, while contribution of CR increases with increasing percentage of deformation. Correspondingly, the size and quantities of  decreased with increasing deformation level. In CR specimens, a high density of nano/micro twins were generated with the twin planes oriented perpendicular to the RD-ND surface. The preferred rotation of the crystals generated the gradient of orientation and internal features of the large austenitic grains. The cross-slip of screw dislocations induced a complete β-fibre, consisting of {110}, {112}, and{123}, with doubled intensity at higher deformation. In addition, the specimens tended to be highly susceptible to the twinning-induced orientation emerged by a predominant ⫽ND. An incomplete but intense α-fibre clustered between {110} and {110} was characterized in the as-deformed condition. Besides achieving highest strength, the current work demonstrates the effects of CR on the material strength without the complex influences of  precipitate.

We have investigated the canine of olive flounder in terms of the nonlinear optical signals of two-photon excitation autofluorescence (2PEF) and second harmonic generation (SHG), whereby two kinds of nonlinear microscope images of the canine were obtained using ~140 femtosecond pulses. For increasing excitation power of laser pulses, a photobleaching effect was found to be significant in the red and green ranges of autofluorescence, where a double-exponential decay of the intensity occurs within hundreds of second due to collagen fluorophores in the canine. In the blue range of autofluorescence, the photobleaching is suppressed due to an energy conversion of the incident laser pulse to SHG signal. As a result, the signal in blue range remains constant for long term exposure to laser. We have also found that the SHG intensity depends on the orienta-tion of linearly polarized excitation, and the optical anisotropy of SHG can be attributed to the collagen alignment of canines.

Abstract: Toxicity of emerging organic pollutants to photosystems of aquatic plants are still not well clarified. This study aimed to develop a novel ecotoxicological experimental protocol based on nanoscale electrochemical mapping of photosynthetic oxygen evolution of aquatic plants by SECM (scanning electrochemical microscopy). The protocol was also checked by confocal laser scanning microscopy (CLSM), the traditional Clark oxygen electrode method and chlorophyll fluorescence technique. The typical persistent organic pollutant 2,4-dichlorophenol (2,4-DCP) in water environment and the common aquatic Microsorium pteropus (M. pteropus) were chosen as the model organic pollutant and tested plant, respectively. It was found that the SECM method could well discriminated the responses of stoma micromorphology and spatial pattens of photosynthetic oxygen evolution on single stoma. The shape of stoma blurred with increasing 2,4-DCP concentration, which was in good agreement with the CLSM images. The dose-response curves and IC50 values obtained from the SECM data were verified by the data measured by the traditional Clark oxygen electrode method and chlorophyll fluorescence test. The IC50 value of single stoma oxygen emission of 24 h exposed plant leave, which was derived from the SECM current data (32535 μg L-1), was close to those calculated from the maximum photosynthetic efficiency (Fv/Fm) measured by chlorophyll fluorescence test (33963 μg L-1), and the Clark oxygen electrode method photosynthetic oxygen evolution rate (32375 μg L-1). The 72 h and 96 h 2,4-DCP exposure data further confirmed the reliability of the nanoscale stoma oxygen emission mapping methodology for ecotoxicological assessment. In this protocol, the procedures for how to collect effective electrochemical data and how to extract useful information from the single stoma oxygen emission pattern were well established. This study showed that SECM is a feasible and reliable ecotoxicological tool for evaluation of toxicity of organic pollutants to higher plants with unique nanoscale visualization advantage over the conventional methods.

This study examines the effects of the δ phase and annealing twins on the hardness, Charpy impact toughness, and tensile properties of Inconel 718 fabricated using L-PBF. Two heat treatment methods, involving solid solution and precipitation hardening, were utilized. The δ phase was formed through solid solution heat treatment at 980C for 1 hour while annealing twins were generated at 1100C for 3 hours. After precipitation hardening, specimens with a higher concentration of  precipitates exhibited increased ultimate tensile strength (13%), yield strength (27%), and hardness (12%) compared to those with annealing twins. This can be attributed to the smaller grain size, subgrains, and δ precipitates, which contribute to the differences in mechanical strength. On the contrary, specimens with annealing twins showed significantly higher impact toughness (4 times) and ductility (twice) than those with δ precipitates. This is caused by the formation of  precipitates, which serve as sites for void formation and crack propagation, leading to a decrease in plasticity due to lattice mismatches with the matrix. Microstructural, compositional, phase, crystallographic, and fractographic analyzes were performed using OM, SEM, TEM and XRD techniques to identify the factors responsible for the variations in the measurements. The results demonstrate that the heat treatment method involving annealing twins significantly improves the ductility of Inconel 718 while maintaining the required mechanical strength.

This study explores the use of remote ultrasound vibration to enhance welding strength. It examines the effects of 20 kHz, and 40 kHz High Power Ultrasound Transducers (HPUT) in contact and contactless modes during ultrasonic assistance laser welding. Employing shear tests and microscopy for assessment, the research evaluates ultrasound's impact on welding strength and identifies the optimal transducer. Theoretical and numerical analyses investigate vibration transmission into the welding area, particularly examining the vibration distribution and cavitation initiation within the molten pool area. Results indicate that ultrasound-assisted welding increases pull test performance, including the resistance force and strain, and improves microscopic structural integrity. Numerical simulations suggest that narrow-band vibration intensifies pressure in specific welding zones. Contactless ultrasound application also shows enhanced welding quality, offering a promising alternative to current methods. This work contributes to electric car technology by suggesting improvements in welding techniques for power battery systems.

A 23-month-old neutered male dog of unknown ancestry presented with a history of progressive neurological signs that included anxiety, cognitive impairment, tremors, seizure activity, ataxia, and pronounced visual impairment. The clinical signs were accompanied by global brain atrophy. Due to progression in the severity of disease signs, the dog was euthanized at 26 months of age. Examination of tissues collected at necropsy revealed dramatic intracellular accumulations of autofluorescent inclusions in the brain, retina, and cardiac muscle. The inclusions were immunopositive for subunit c of mitochondrial ATP synthase, and their ultrastructural appearances were similar to those of lysosomal storage bodies that accumulate in some of the neuronal ceroid lipofuscinosis (NCL) diseases. The also dog exhibited widespread neuroinflammation. Based on these findings, the dog was deemed likely to have suffered from a form of NCL. A whole genome sequence analysis of the proband’s DNA revealed a homozygous C to T substitution that altered the intron 3 – exon 4 splice site of CLN6. Other mutations in CLN6 cause NCL diseases in humans and animals including dogs. CLN6 protein was undetectable with immunolabeling in the tissues of the proband. Based on the clinical history, fluorescence and electron-microscopy, immunohistochemistry, and molecular genetic findings, the disorder in this dog is classified as an NCL resulting from the absence of CLN6 protein. Screening the dog’s genome for a panel of breed-specific polymorphisms indicated that its ancestry included numerous breeds, with no single breed predominating. This suggests that the CLN6 disease variant is likely to be present in other mixed breed dogs and at least some of the ancestral breeds.

Abstract Cerebral microbleed(s) (CMBs) are increasingly being viewed not only as a marker for cerebral small vessel disease (SVD) but also as having an increased risk for the development of stroke (hemorrhagic/ischemic) and aging-related dementia. Recently, brain endothelial cell activation and dysfunction and blood-brain barrier dysfunction and/or disruption have been shown to be associated with SVD, enlarged perivascular spaces, and the development and evolution of CMBs. CMBs are a known disorder of cerebral microvessels that are visualized as 3-5mm, smooth, round or oval, and hypointense (black) lesions seen only on T2*-weighted gradient recall echo or susceptibility-weighted sequences on MRI images. CMBs (a global problem) are known to occur with high prevalence in community-dwelling older individuals. Since our current global population is the oldest recorded in history and only expected to continue to grow, we can only expect the health care burdens associated with CMBs to also grow. CMBs should raise a red flag regarding the increased risk of large symptomatic neurologic intracerebral hemorrhages. Importantly, CMBs are also currently regarded as markers of diffuse vascular and neurodegenerative brain damage. Thus, it is essential that we try to learn as much as we can about their development, evolution, and their relation to impaired cognition, dementia, and neurodegeneration.

Cell mechanics is a biophysical indicator of cell state, such as cancer metastasis, leukocyte activation, and cell cycle progression. Atomic force microscopy (AFM) is a widely used technique to measure cell mechanics, where the Young modulus of a cell is usually derived from the Hertz contact model. However, the Hertz model assumes that the cell is an elastic, isotropic, and homogeneous material and that the indentation is small compared to the cell size. These assumptions neglect the effects of the cytoskeleton, cell size and shape, and cell environment on cell deformation. In this study, we investigated the influence of cell size on the estimated Young’s modulus using liposomes as cell models. Liposomes were prepared with different sizes and filled with phosphate buffered saline (PBS) or hyaluronic acid (HA) to mimic the cytoplasm. AFM was used to obtain the force indentation curves and fit them to the Hertz model. We found that the larger the liposome, the lower the estimated Young’s modulus for both PBS-filled and HA-filled liposomes. This suggests that the Young modulus obtained from the Hertz model is not only a property of the cell material, but also depends on the cell dimensions. Therefore, when comparing or interpreting cell mechanics using the Hertz model, it is essential to account for cell size.