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.

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.

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.

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.

Theoretical and experimental approaches have been applied to study the polymer physics under-lying compaction of DNA in the bacterial nucleoid. Knowledge of the compaction mechanism is necessary to obtain a mechanistic understanding of the process of segregation of replicating chromosome arms (replichores) during the cell cycle. In the first part of this review light microscope observations are discussed that have demonstrated that the nucleoid has a lower refractive index and thus a lower density than the cytoplasm. A polymer-physical explanation for this phenomenon was given by the theory of Theo Odijk pro-posed in 1998. By assuming a phase separation between nucleoid and cytoplasm and by imposing equality of osmotic pressure and chemical potential between the two phases, a minimal energy situation is obtained in which soluble proteins are depleted from the nucleoid, thus explaining its lower density. The theory is compared with recent views for DNA compaction that are based on mere exclusion of polyribosomes from the nucleoid or on the transcriptional activity of the cell. These new views raise the question of whether they can still explain the lower refractive index or density of the nucleoid. In the second part of this review the question is discussed how DNA segregation occurs in Esche-richia coli, in the absence of the so-called active ParABS system, present in the majority of bacteria. How is entanglement prevented of nascent chromosome arms, generated at the origin in the pa-rental network of the E. coli nucleoid? The observations of the groups of Sherratt and Hansen in 2006, that the four nascent chromosome arms synthesized in the initial replication bubble, segre-gate to opposite halves of the sister nucleoids implies that extensive intermingling of daughter strands does not occur. Based on the hypothesis that leading and lagging replichores synthesized in the replication bubble, fold into micro-domains that do not intermingle, a passive "four-excluding-arms model" for segregation is proposed. This model implies that the key for seg-regation already lies in the structure of the replication bubble at the very start of DNA replication; it explains the different patterns of chromosome arms as well as the segregation distances between replicated loci as experimentally observed.

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.

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.

Sub-micro dislocation cellular structures formed during rapid solidification break the strength-ductility trade-off in laser powder bed fusion (LPBF)-processed 316L stainless steel through high-density dislocations and segregated elements or precipitates at the cellular boundaries. The high-density dislocation entangled at the cellular boundary accommodates solidification strains among the cellular structures and cooling stresses through elastoplastic deformation. Columnar grains with cellular structures typically form along the direction of thermal flux. However, the ultra-low misorientations between the adjacent cellular structures and their interactions with the cellular boundary formation remain unclear. In this study, we revealed the ultra-low misorientations between the cellular structures in LPBF-processed 316L stainless steel using conventional electron backscatter diffraction (EBSD), transmission Kikuchi diffraction (TKD), and transmission electron microscopy (TEM). The conventional EBSD and TKD analysis results could provide the misorientation angles smaller than 2° while the resolution mainly depends on the specimen quality and scanning step size, and so on. TEM technique with higher spatial resolution provides accurate information between adjacent dislocation cells with misorientation angles smaller than 1°. This study presents that TEM method is the better and more precise analysis method of the misorientation measurement of the cellular structures and provides insights into measuring the small misorientation angles between adjacent dislocation cells and nanograins in nanostructured-metals and alloys with ultrafine-grained microstructures.

Axial resolution is one of the most important characteristics of a microscope. In all microscopes, a high axial resolution is desired in order to discriminate information efficiently along the longitudinal direction. However, when studying thick samples that do not contain laterally overlapping information, a low axial resolution is desirable as information from multiple planes can be recorded simultaneously from a single camera shot instead of plane-by-plane mechanical refocusing. In this study, we increased the focal depth of an infrared microscope non-invasively by introducing a binary axicon fabricated on barium fluoride substrate close to the sample. Preliminary results of imaging thick and sparse silk fibers showed an improved focal depth with a slight decrease in lateral resolution and increase in background noise.

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.

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.

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.

This study aimed to evaluate the impact of Candida albicans on subgingival biofilm formation on dental implant surfaces. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to compare biofilm structure and microbial biomass in the presence and absence of the fungus after periods of 24, 48, and 72 h. Quantitative polymerase chain reaction (qPCR) was used to quantify the number of viable and total micro-organisms for each of the biofilm-forming strains. A general linear model was applied to compare CLSM and qPCR results between control and test conditions. The biofilm developed with C. albicans at 72 h had a higher bacterial biomass, with a significantly higher cell viability (p < 0.05). After both 48 and 72 h of incubation, in the presence of C. albicans, there was a significant increase in counts of Fusobacterium nucleatum and Porphyromonas gingivalis and in the cell viability of Streptococcus oralis, Aggregatibacter actinomycetemcomitans, F. nucleatum, and P. gingivalis. Using a dynamic in vitro multispecies biofilm model, C. albicans exacerbated the development of the biofilm grown on dental implant surfaces, significantly increasing the number and cell viability of periodontal bacteria.

The up-to-date challenge towards clean road transport are non-exhaust emissions. Important advances regarding measurement systems, including well defined characterisation techniques, as well as regulation, will happen in the next few years. In this work we present detailed results of particle emissions from a test bed for wheel suspension and brakes, consisting of aerosol (size distribution, particle number (PN) and mass (PM)) and electron microscopy measurements (EM) under different load conditions. Standard tyres and brakes from series production were tested with a high-load driving cycle while particle measurements were conducted by gravimetric measurements and with a TSI SMPS, a TSI APS and a GRIMM OPS. Furthermore, samples were analyzed by electron microscopy. A bimodal particle size distribution (PSD) was obtained with the SMPS, with peaks at 20nm and around 400nm. EM analysis of >1400 single particles from the electrostatic sampler matches the PSD results. EM analysis also showed ultrafine particles, mainly from O, Fe, Si, Ba, Mg and S and also fractal particles with high C fractions. Our results suggest, in agreement with previously published literature, that particulate emissions are related to brake disc temperature and occur in significant amounts above a threshold temperature.

Primary ciliary dyskinesia (PCD) is an inherited disorder that impairs motile cilia, essential for respiratory health, with a reported prevalence of 1 in 16,309 within Hispanic populations. Despite having the RSPH4A [c.921+3_921+6del (intronic)] founder mutation in 70% of Puerto Rican patients, the characterization of the ciliary dysfunction remains unidentified due to the unavailability of advanced diagnostic modalities like High-Speed Video Microscopy Analysis (HSVA). Our study implemented HSVA for the first time on the island as a tool to better diagnose and characterize the RSPH4A [c.921+3_921+6del (intronic)] founder mutation in Puerto Rican patients. By applying HSVA, we analyzed the ciliary beat frequency (CBF) and pattern (CBP) in native Puerto Rican patients with PCD. Our results showed decreased CBF and a rotational CBP linked to the RSPH4A founder mutation in Puerto Ricans, presenting a novel diagnostic marker that could be implemented as an axillary test into the PCD diagnosis algorithm in Puerto Rico. The integration of HSVA technology in Puerto Rico substantially enhances the framework for PCD evaluation and diagnosis, facilitating prompt detection and early intervention for improved disease management. This initiative, demonstrating the potential of HSVA as an adjunctive test within the PCD diagnostic algorithm, could serve as a blueprint for analogous developments throughout Latin America.

Chitosan (CS) nanoparticles are under investigation for a wide range of applications in nanomedicine. The structural, molecular and physicochemical properties of chitosan nanoparticles have a direct bearing on their efficacy in drug conjugation and delivery, gene delivery, and tissue engineering. Herein we report on the structural, morphological and molecular properties of chitosan nanoparticles synthesized using the ionic gelation method with the intended purpose of being used for drug delivery. The chitosan nanoparticles (CS-NPs) were prepared in the 0.25 to 1.0% w/v concentration range, whereas separate samples of the 1.0% w/v CS-NPs were also functionalized with polyethylene oxide (PEO) alone and with a diblock copolymer of PEO and polypropylene glycol (PPG). The average size of the CS-NPs was determined from TEM imaging to be in the range of 11.3 to 14.8 nm. The XRD and TEM analyses show that the CS-NPs have semi-crystalline structure with the degree of crystallinity depending upon the nature of synthesis. Our results show that functionalizing the 1.0% w/v CS-NPs with PEO increases the degree of crystallinity whereas functionalizing with the diblock copolymer PEO - PPG results in a significant increase in their degree crystallinity. The CS/TPP concentration (CS:TPP fixed at 1:1 ratio) had no apparent effect on the degree of crystallinity of the CS-NPs. The results from the FTIR analysis confirm incorporation of TPP with CS in the CS-NPs. Additionally, FTIR analysis shows an enhancement of hydrogen bonding that is consistent with the increase in the degree of crystallinity of the CS-NPs.

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.

The present paper explored the possibility of discriminating between different malignant degrees of Chondrosarcoma (CS), a diffused bone tumor, and Enchondroma (EC), its benign version, through an approach based on the coupling between Confocal Raman Microscopy (CRM) and Machine Learning (ML) techniques. Recently, Raman Spectroscopy has proven to be a powerful tool for a complete grading of CS by distinguishing between grade 1, 2 and 3, and to distinguish CS from EC. In this paper, we tested some models, belonging either to ML or to the sub-type called Deep Learning (DL), showing excellent classification performances, especially the DL algorithms, with classification accuracy approaching to 100%, making the models promising for future implementations of Raman spectroscopy when applied to oncology diagnosis. In turn, we highlighted how some proper ML models result in worse classification performances but better resolution of specific chemical target compounds, possible candidate to become malignant markers, with relevant implication for a correct diagnosis.

(1) Background: Myelodysplastic syndromes (MDS) are a group of diseases that affect the morphology and function of neutrophils. Rapid diagnosis of MDS is crucial for the initiation of treatment that can vastly improve disease outcome. (2) methods: In this work, we present a new approach for detecting morphological differences between neutrophils isolated from blood samples of high risk MDS patients and that for patients and blood bank donors (BBDs). First, using fluorescent flow cytometry CD66b positive cells were stained with 2′,7′-dichlorofluorescin diacetate (DCF) and Hoechst. Secondly, we imaged the cells using interferometric phase microscopy (IPM) without utilizing cell staining. (3) Results: flow cytometry results show MDS patients present different dispersion patterns compared to BBDs. BBDs present two distinct subpopulations (one of these are the H cells constituting 16% for neutrophils) whereas MDS patients present only 4% of this subpopulation. IPM results are consistent with this pattern: ~20% of BBDs neutrophils were characterized as H cells and whereas MDS patients present only ~4% of these cells. (5) Conclusion- IPM can be utilized for diagnosis of complex hematological pathologies such as MDS.

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.

Quantitative phase imaging by digital holographic microscopy (DHM) is a nondestructive and label-free technique, that has been playing an indispensable role in the field of science, technology, and biomedical imaging. The technique is competent for the imaging and analysis of label-free living cells and the investigation of reflective surfaces. Herein, we introduce a new configuration of a wide field-of-view single-shot common-path off-axis reflective DHM for the quantitative phase imaging of biological cells that leverages with several advantages including less-vibration sensitive to external perturbations due to common-path configuration, compact in size, simple in optical design, highly stable, and cost-effective. A detailed description of the proposed DHM system, including its optical design, working principle, and capability of phase imaging, is presented. The applications of the proposed system are demonstrated by quantitative phase imaging results obtained from the reflective surface (USAF resolution test target) as well as transparent samples (microlens array and living plant cells). The proposed system could find its applications in the investigations of several biological specimens and optical metrology of micro-surfaces.

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.

Spatial-structured longitudinal light beams are optical fields sculpted in three-dimensional (3D) space by diffractive optical elements. These beams have been recently suggested for use in im-proving several imaging capabilities, such as 3D imaging, enhancing image resolution, engineering the depth of field, and sectioning 3D scenes. All these imaging tasks are performed using coded aperture correlation holography systems. Each system designed for a specific application is characterized by a point spread function of a different spatial-structured longitudinal light beam. This article reviews the topic of applying certain structured light beams for optical imaging.

Dendritic spines are essential for synaptic function because they constitute the neurons' postsynaptic compartment that receives the most excitatory input. The extracellularly shorter variant of the presynaptic cell adhesion molecules neurexins, β-neurexins, has been implicated in various aspects of synapse function including neurotransmitter release. However, its role in developing or stabilizing dendritic spines as fundamental computational units of excitatory synapses remained unclear. Here, we show by morphological analysis that the deletion of β-neurexins in hippocampal neurons in vitro and in hippocampal tissue in vivo affects presynaptic dense-core vesicles and, unexpectedly, postsynaptic spine structure. Specifically, we observed that the absence of β-neurexins led to an increase in longer spinous protrusions in vitro and more mature mushroom-type spines in the CA1 region of adult knockout mice. In addition, deletion of β-neurexins caused alterations in spine head dimension and more spines with perforations of their postsynaptic density but no change in the overall number of spines or synapses. Our results indicate that presynaptic β-neurexins play a role across the synaptic cleft, possibly by aligning with postsynaptic binding partners and glutamate receptors via transsynaptic columns.

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.

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.

ATP-binding cassette (ABC) transporters, acting as cellular "pumps," facilitate solutes translocation through membranes via ATP hydrolysis. Their overexpression is closely tied to multidrug resistance (MDR), a major obstacle in chemotherapy and neurological disorder treatment, hampering drug accumulation and delivery. Extensive research has delved into the intricate interplay between ABC transporter structure, function, and potential inhibition for MDR reversal. Cryo-electron microscopy has been instrumental in unveiling structural details of various MDR-causing ABC transporters, encompassing ABCB1, ABCC1, ABCG2, as well as recently revealed ABCC3 and ABCC4 structures. The newly obtained structural insight has deepened our understanding of substrate and drug binding, translocation mechanisms, and inhibitor interactions. Given the growing body of structural information available for human MDR transporters and their associated mechanisms, we believe it is timely to compile a comprehensive review of these transporters and compare their functional mechanisms in the context of multidrug resistance. Therefore, this review primarily focuses on the structural aspects of clinically significant human ABC transporters linked to MDR, with the aim of providing valuable insights to enhance the effectiveness of MDR reversal strategies in clinical therapies.

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.

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 lack of accurate and feasible diagnostic tests poses a significant challenge to visceral leish-maniasis (VL) healthcare services in endemic areas. To date, various VL diagnostic tests have been or are being developed, and their diagnostic performance needs to be assessed.. In the present study, the diagnostic performance of rk39RDT, Direct Agglutination Test (DAT), Microscopy, Loop Mediated Isothermal Amplification (LAMP), and miniature direct-on-blood polymerase chain reaction - nucleic acid lateral flow immunoassay (mini-dbPCR-NALFIA) was assessed using quantitative polymerase chain reaction (qPCR) as reference test in an endemic region of Ethiopia. In this study, 235 patients VL suspected patients and 104 non-endemic healthy controls (NEHC) were recruited. Among the VL-suspected patients, 144(61.28%) tested positive with qPCR. Sensi-tivities for rk39 RDT, DAT, Microscopy, LAMP assay, and mini-dbPCR-NALFIA were 88.11%, 96.50%, 76.58%, 94.33%, and 95.80%, respectively. Specificities were 83.33%, 97.96%, 100%, 97.38%, and 98.92% for rk39 RDT, DAT, Microscopy, LAMP assay, and mini-dbPCR-NALFIA, respectively. In conclusion, Rk39 RDT and microscopy exhibited lower sensitivity, while DAT demonstrated excellent performance. LAMP and mini-dbPCR-NALFIA showed excellent perfor-mance with feasibility to implement in remote endemic areas, although the latter requires further evaluation in such regions.

Malaria is rare in Europe, but it is a medical emergency and programs for its control should ensure access to early diagnosis and prompt treatment within 24-48 h of the symptom’s onset. The consistent number of imported malaria cases and the risk of reappearance of autochthonous cases stimulated laboratories in non-endemic countries to evaluate diagnostic methods/algorithms: microscopy remains the gold standard, but with limitations. RDTs have greatly expanded the ability to diagnose malaria for rapid results due to simplicity and low cost, but they lack sensitivity and specificity. PCR-based assays provide more relevant information but need well-trained technicians. According to WHO global Technical Strategy for Malaria 2016-2030, the future direction for a rapid diagnosis is the development of Point-Of-Care Testing, which would be helpful for prompt/accurate treatment and for preventing the diseases spread. Despite the limitations of current diagnostic methods, they play important roles in dealing with the global malaria situation, including decreasing its incidence. Recently evidence suggested that Artificial Intelligence could be utilized for assisting pathologists in malaria diagnosis. Future studies need to be conducted to see whether this system can be used for routine malaria diagnostic procedures.

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.

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.

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.

Smelting used to be less efficient in the past, so wastes obtained from historical processing at smelter plants usually contain certain quantities of valuable metals. A case study was conducted, which included an investigation of the physicochemical, mineralogical, and microstructural properties of raw material found at the historic slag pond near the Topilnica Veles smelter in North Macedonia. The Pb-Zn slag was sampled using traditional geophysical methods such as drill holes and pitholes. 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 (2.24 wt%), Zn (7.10 wt%), and Ag (27.53 ppm). Amorphous phase, lead alloys, zinc alloys, wurtzite, sphalerite, galena, cerussite, elemental silver, elemental copper, elemental iron, magnetite, spinel, rutile, hematite, and troilite were also identified through qualitative mineralogical analysis. The glassy matrix composed of spinel, silicates, and mixed spinel-silicate is the most abundant constituent in slag composition. The various potential applications of the slag are possible based on its mineralogical and geochemical properties. Because the extraction of Pb, Zn, and Ag has certain economic potential, the next step, which will include gravity concentration and magnetic separation procedures, will be to form metal concentrates into their collective concentrate, from which the maximum amount of silver can be extracted. Slag's amorphous spinel, silicate, and mixed spinel-silicate phases, which contain high concentrations of SiO2, Al2O3, CaO, and Fe2O3, are suitable for use in building materials such as cement clinker, filler, or aggregate for concrete or mortar. The goal of this research is to close the circle of slag's reutilization potential in terms of Net Zero and 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.

It remains unclear in the literature what is the cause of the so-called alkali-carbonate reaction (ACR) damage to concrete yet, a consensus is reached when considering dedolimitization as non-expansive. However, expansion and cracks as distress features are often attributed to alkali-silica reaction (ASR) when evidence of a reactive mineral is found in the aggregate. Evidently, different types of aggregates may produce different types and degrees of damage. Therefore, this work aims to assess the damage to concrete generated and propagated by the so-called ACR susceptible reactive Kingston coarse aggregate through mechanical testing (i.e., direct shear test), microscopy (the damage rating index – DRI), and other techniques. Distinct induced expansion levels (i.e., 0%, 0.05%, 0.12%, and 0.20%) were selected to compare the distress caused by ACR to concrete affected by ASR. Result show that the behaviour of ACR, namely as captured through the DRI, is inconsistent with that of ASR thus attesting to ACR being a distinct distress mechanism.

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.

Copper-nitride (Cu3N) semiconductor material is attracting much attention as a potential, next-generation thin-film solar-light absorber in solar cells. In this communication, polycrystalline Cu3N thin films were prepared using reactive-RF-magnetron-sputtering deposition, at room temperature, onto glass and silicon substrates. The very-broadband optical properties of the Cu3N thin film layers were studied by UV-MIR (0.2-40 μm) ellipsometry and optical transmission, to be able to achieve the goal of a low-cost absorber material to replace the conventional silicon. The reactive-RF-sputtered Cu3N films were also investigated by focused ion beam scanning electron microscopy, and both FTIR and Raman spectroscopies.

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.

Polyvinyl Alcohol (PVA)/ Graphene Quantum Dots (GQDs) polymer nanocomposite films have been prepared, and the relationship between their structural, thermal and nanoscale morphologi-cal properties, and their photoluminescent response, has been investigated. Although according to X-ray Diffraction (XRD), Fourier-Transform Infrared Spectroscopy (FT-IR) and Differential Thermal Analysis (DTA) the incorporation of GQDs does not significantly affect the percentage crystallinity of the PVA matrix, for a range of added GQD concentrations, Atomic Force Micros-copy (AFM) showed the formation of islands with apparent crystalline morphology on the surface of the PVA/GQD films, with GQD presumably acting as a nucleating agent for island growth. The incorporation of GQD also led to the formation of characteristic surface pores with increased stiffness and frictional contrast, according to Ultrasonic Force Microscopy (UFM) and Frictional Force Microscopy (FFM) data. The photoluminescence (PL) spectra of the films were found to de-pend both on the amount of GQD incorporated and on the film morphology. For GQD loads > 1,2wt%, a GQD-related band was observed at ~ 1650 cm-1 in FT-IR, along with an increase in the PL band at lower energy. For a load of ~2wt% GQD, the surface morphology was characterized by extended cluster aggregates with lower stiffness and friction than the surrounding matrix, and the PL signal decreased.

Cryo electron microscopy (cryo-EM) instrumentation allows obtaining 3D reconstruction of the structure of biomolecular complexes in vitro (purified complexes studied by single particle analysis) and in situ (complexes studied in cells by cryo electron tomography). Standard cryo-EM approaches allow high-resolution reconstruction of only a few conformational states of a molecular complex, as they rely on data classification into a given number of classes to increase the resolution of the reconstruction from a few, most populated classes and discard all other classes. Such discrete-classification approaches result in a partial picture of the full conformational variability of the complex, due to continuous conformational transitions with many, uncountable intermediate states. In this article, we present the software with user-friendly graphical interface for running two recently introduced methods, namely MDSPACE and MDTOMO, to obtain continuous conformational landscapes of biomolecules by analyzing in vitro and in situ cryo-EM data (single particle images and subtomograms) based on molecular dynamics simulations of an available atomic model of one of the conformations. The MDSPACE and MDTOMO software is part of the open-source ContinuousFlex package, which can be run as a plugin of the Scipion software package, broadly used in the cryo-EM field.

Scanning electron microscopy (SEM) is a powerful tool in the domain of material science, mining, and geology, owing to its enormous potential to provide unique insights into the micro and nanoscale worlds. This comprehensive review discusses the background development of SEM, basic SEM operation including the specimen preparation and imaging process, and fundamental theoretical calculations underlying the SEM operation. It provides foundational understanding to the engineers and scientists, who never got a chance to dig in-depth into the SEM, to understand the working and development of this robust analytical technique. The present review covers how SEM has been serving as a crucial tool in mineral characterization, with specific discussions on the working and research fronts of SEM-EDX, SEM-AM, SEM-MLA, and QEMSCAN. With automation gaining pace in the development of all spheres of technology, the understanding of uncertainties in SEM measurements is very important. The constraints in mineral phase identification by EDS spectra and sample preparation are conferred. In the end, future research directions for SEM are analyzed with the possible incorporation of machine learning, deep learning, and artificial intelligence tools, for automating the process of mineral identification, quantification, and efficient communication with the researchers, so that the analytical process robustness and objectivity can be improved, and the analysis time and the involved costs can be brought down. This review also discusses the idea of integrating robotics with SEM, to make the equipment portable, so that further mineral characterization insights can be gained not only on earth but also on other terrestrial grounds.

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.

The first hyperpolarizability of graphene quantum dots (GQDs) suspended in water was determined using the hyper-Rayleigh scattering (HRS) technique. To the best of our knowledge, this is the first application of the HRS technique to characterize GQDs. Two commercial GQDs (Acqua-Cyan and Acqua-Green) with different compositions were studied. The HRS experiments were performed with an excitation laser at 1064 nm. The measured hyperpolarizabilities were (1.0±0.1) × 10^(-27) esu and (0.9±0.1) × 10^(-27) esu for Acqua-Cyan and Acqua-Green, respectively. The results were used to estimate the hyperpolarizability per nanosheet obtained by assuming that each GQDs has five nanosheets with 0.3 nm thickness. The two-level model, used to calculate the static hyperpolarizability per nanosheet, provides the values of (2.4±0.1) × 10^(-28) esu (Acqua-Cyan) and (0.5±0.1) × 10^(-28) esu (Aqua-Green). The origin of the nonlinearity is discussed on the basis of polarized resolved HRS experiments, and an electric quadrupolar behavior with a strong dependence on surface effects is found. The nontoxic characteristics and order of magnitude indicate that these GQDs may be useful for biological microscopy imaging.

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.

Modern imaging strategies are paramount to studying living systems such as cells, bacteria, and fungi and their response to pathogens, toxicants, and nanomaterials as modulated by exposure and environmental factors. The need to understand the processes and mechanisms of damage, healing and cell survivability of living systems continues to motivate the development of alternative imaging strategies. Of particular interest is the use of label-free techniques that minimize interference of biological processes by foreign marking substances and reduce intense light exposure and potential photo-toxicity effects. This review focus on the potential synergic capabilities of atomic force microscopy (AFM) as a well-developed and robust imaging strategy with demonstrated applications to unravel intimate details in biomedical applications, with the label-free, fast, and enduring Holotomographic Microscopy (HT) strategy. We first review the operating principles that form the basis for the complementary details provided by these techniques regarding the surface and internal information provided by HT and AFM is essential and complimentary for the development of several biomedical areas studying the interaction mechanisms of nanomaterials with living organisms. First, AFM can provide superb resolution on surface morphology and biomechanical characterization. Second, the quantitative phase capabilities of HT enable superb modeling and quantification of the volume, surface area, protein content, and mass density of the main components of cells and microorganisms, including morphology of cells in microbiological systems. These capabilities result from directly quantifying refractive index changes without requiring fluorescent markers or chemicals. As such, HT is ideally suited for long-term monitoring of living organisms in conditions close to their natural settings. We present a case-based review of the principal uses of both techniques and their essential contributions to nanomedicine and nanotoxicology, emphasizing cancer and infectious disease control. The synergic impact of the sequential use of these complementary strategies provides a clear drive for adopting these techniques as interdependent fundamental tools.

Wild-type p53 cancer therapy-induced senescent cells frequently engulf and degrade neighboring ones inside a massive vacuole in their cytoplasm. After clearance of the internalized cell, the vacuole persists, seemingly empty, for several hours. Despite large vacuoles being associated with cell death, this process is known to confer a survival advantage to cancer engulfing cells, leading to therapy resistance and tumor relapse. Previous attempts to resolve the vacuolar structure and visualize their content using dyes were unsatisfying for lack of known targets and ineffective dye penetration and/or retain. Here, we overcame this problem by applying optical diffraction tomography and Raman spectroscopy to MCF7 doxorubicin-induced engulfing cells. We demonstrated a real ability of cell tomography and Raman to phenotype complex microstructures, such as cell-in-cells and vacuoles, and detect chemical species in extremely low concentrations within live cells in a completely label-free fashion. We show that vacuoles had a density indistinguishable to the medium, but were not empty, instead contained diluted cell-derived macromolecules, and we could discern vacuoles from medium and cells using their Raman fingerprint. Our approach is useful for the non-invasive investigation of senescent engulfing (and other peculiar) cells in unperturbed conditions, crucial for a better understanding of complex biological processes.

Criminal investigations aimed to track the route walked by Missing Persons and Fugitives (MPFs) usually involve Intelligence analysts, military planners, experts in mobile forensics, traditional investigative methods, and sniffer dog handlers. Notwithstanding, when MPFs are devoid of any technological device and move in uninhabited rural areas devoid of tele cameras and densely covered by vegetation, tracking backwards the route walked by MPFs may be a much more arduous task. In such complex cases, a very efficient approach may consist in comparing the geological traces found on the MPFs with soils and plants exposed in the event scenes. In particular, the search for peculiar or rare particles and aggregates may strengthen the weight of the geological evidence comparisons. A match of mineralogical, textural, and botanical data may demonstrate the provenance of the traces from the soil of a specific site, linking in this way the MPFs to the scene of events. Based on the above, the present paper reports geological and botanical determinations accomplished for a “mediatic” casework. Results allowed to ascertain a general high degree of compatibility among traces collected on the MPFs and on the soil from the scene of events. The most significant positive matches, based on the finding of a ten of peculiar and rare particles and assemblages, allowed reconstructing a route about 1.1 km long, as the crow flies, on the event site. Notwithstanding this procedure was extremely time consuming and available only in a backwards reconstruction linked to the MPFs’ findings, it was of uttermost importance in strengthen the inferences proposed, where other methods could not provide any information.

Samples of ZrO2 ceramics with different concentrations of impurity titanium ions were synthesized by mixing powders of zirconium and titanium oxides in various mass ratios. The phase composition and surface morphology of the studied ceramics were determined by X-ray phase analysis and scanning electron microscopy. It was found that irradiation of samples with xenon ions (220 MeV) with fluences 1010 and 1012 ion/cm2 leads to a decrease in the intensity of the pulse cathodoluminescence band at 2.5 eV. It is shown that ion irradiation causes the appearance of a new peak of thermoluminescence at 450-650 K of a non-elementary form associated with radiation-induced traps of charge carriers. In contrast to electron irradiation, an increase in the fluence of ions leads to a decrease in the intensity of this peak. In contrast to electron irradiation, an increase in the fluence of ions leads to a decrease in the intensity of this peak. A complex nonmonotonic dependence of the cathode and thermoluminescence intensity on the dopant concentration was found, which may be due to the effects of concentration quenching and aggregation of defective centers.

Severe COVID-19 alters the biochemical and morphological characteristics of blood cells in a wide variety of ways. To date, however, the vast majority of research has been devoted to the study of leukocytes, while erythrocyte morphological changes have received significantly less attention. The purpose of this research was to identify erythrocyte types that were unique to COVID-19, compare the number of different poikilocyte types, and measure erythrocyte sizes to provide data on size dispersion. Red blood cells obtained from 6 control donors (800-2200 cells for each donor) and 5 COVID-19 patients (800-1900 cells for each patient) were examined using low voltage scanning electron microscopy. We did not discover any forms of poikilocytes that would be unique to COVID-19. Among COVID-19 patients, we observed an increase in the number of acanthocytes (p=0.01) and a decrease in the number of spherocytes (p=0.03). In addition, our research demonstrates that COVID-19 causes an increase in the median (p=0.004) and interquartile range (p=0.009) when assessing erythrocyte size.

The introduction on Zinc oxide nanoparticles (ZnOn) in sunscreens solved the issue of poor spreadability of these formulations, which often left a white film on the skin. However, safety concerns have arisen regarding the topical application of ZnOn. Some studies employed commercial sunscreens to address the safety issues of the topical application of ZnOn; however, commercial formulations are often complex and contain a wide range of ingredients that could attenuate the potential damage caused by the ZnOn. Therefore, in this study we aimed to develop a simple stable formulation containing 20% of coated and uncoated ZnOn, characterize the formulations and the nanoparticles, and assess the skin penetration in Franz diffusion cell. The Feret's diameter for the uncoated and coated ZnOn was 137 nm and 134 nm, respectively. For the uncoated ZnOn the hydrodynamic size in water was 368 nm and for the coated ZnOn, the average hydrodynamic size in ethyl acetate was 135 nm. The incorporation of ZnOn led to formulations more consistent and easier to spread, as suggested by the lower work of shear and higher values of firmness, cohesiveness, consistency and index of viscosity compared with the vehicle. The stability assessment at 45ºC suggested that the formulations containing the ZnOn were stable for 30 days and the vehicle was stable for 90 days. The assessment off the skin penetration by reflectance confocal laser microscopy indicated that the formulations did not permeate into the deepest layers of the skin, but accumulated on the skin furrows, hair and hair follicle.

This study presents a combined electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM) and Scanning Kelvin Probe Force Microscopy(SKPFM) investigation on the natural passivation process of stainless steels (AISI 316 and AISI 304) and nickel-based alloy (INCONEL 600) as a function of immersion time in an alkaline medium (pH = 13). EIS measurements show an increase in the resistance of the film with increasing immersion time in the electrolyte. After 3 days of immersion, for the three alloys, the films formed constitute an effective barrier to the dissolution of the alloys as shown by AFM and SKPFM. The oxide film growth on each substrates is only influenced by trenches formed during the polishing step and don’t depend on the chemical composition.

The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced and 599 candidate genes exhibited differential expression between DSG3-overexpressing and matched control lines, with 12 genes highly significantly differentially expressed, including 9 upregulated and 3 downregulated. Genes with known implications in cancer, such as MMP-13, KRT84, OLFM4, GJA1, AMOT, and ADAMTS1, were strongly linked to DSG3 overexpression. Gene ontology analysis indicated that the DSG3-associated candidate gene products participated in crucial cellular processes such as junction assembly, focal adhesion, extracellular matrix formation, intermediate filament organization, and keratinocyte differentiation. Validation of RNA-Seq was performed through qRT-PCR, Western blotting, and immunofluorescence analyses. Furthermore, transmission electron microscopy meticulously examined desmosome morphology, revealing slightly immature desmosome structure in DSG3-overexpressing cells compared to controls. No changes in desmosome frequency and diameter were observed between the two conditions. This study underscores intricate and multifaceted alterations associated with DSG3 in oral squamous carcinoma cells, implying a potential oncogenic role of this gene in biological processes that enable cell communication, motility and survival.

Single-particle cryo-electron microscopy (cryo-EM SPA) has recently emerged as an exceptionally well-suited technique for determining the structure of membrane proteins (MPs). Indeed, in the last years, it was observed a huge increase in the number of MPs solved by cryo-EM SPA at a resolution better than 3.0 Å in the Protein Data Bank (PDB). However, sample preparation remains a significant challenge in the field. Here, we evaluated in the MPs solved by cryo-EM SPA deposited in the PDB in the last two years at a resolution below 3.0 Å the most critical parameters for sample preparation: i) the surfactant used for protein extraction from the membrane, ii) the surfactant, amphiphiles, nanodiscs or other molecules present in the vitrification step, iii) the vitrification method employed, and iv) the type of grids used. The aim is not to provide a definitive answer on the optimal sample conditions for cryo-EM SPA of MPs, but rather assess the current trends in the MP structural biology community towards obtaining high-resolution cryo-EM structures.

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.

Concatemeric viral DNA is packaged into Bacteriophage P22 procapsids via a headful packaging mechanism mediated by a molecular machine consisting of small (gp3) and large (gp2) terminase subunits. Although a negative stain reconstruction exists for the terminase holoenzyme, it is not clear how this complex binds dodecameric portal protein located at a 5-fold mismatch vertex. Herein, we describe new assemblies for the holoenzyme. Both native mass spectrometry and transmission electron microscopy reveal that the P22 terminase complex adopts three main assemblies, which include a nonameric S-terminase bound to two L-terminase 1(gp3)9:2(gp2), two nonameric S-terminase bound to five L-terminase 2(gp3)9:5(gp2), and three nonameric S-terminase bound to seven L-terminase 3(gp3)9:7(gp2). Native agarose gel electrophoresis shows that the terminase complex interacts with procapsid with mild crosslinking. These results herein illustrate the P22 terminase complex can adopt a variety of confirmations and assembly states.

The phenomenon of ordered self-assembly on solid substrates is a topic of interest in both fundamental surface science research and its applications in nanotechnology. The regulation and control of two-dimensional (2D) self-assembled supra-molecular structures on surfaces have been realized through applying external stimuli. By utilizing scanning tunneling microscopy (STM), researchers can investigate detailed phase transition process about self-assembled monolayers (SAMs), providing insight into the interplay between intermolecular weak interactions and substrate-molecule interactions, which govern the formation of molecular self-assembly. This review will discuss the structural transition of self-assembly probed by STM in response to external stimuli and provide state-of-art methods such as tip induced confinement for the alignment of SAM domains and selective chirality. Finally, we discuss the challenges and opportunities in the field of self-assembly and STM.

Striated muscle thick filaments are composed of myosin II and several non- myosin proteins which define the filament length and modify its function. Myosin II has a globular N-terminal motor domain comprising its catalytic and actin-binding activities and a long α-helical, coiled-coil tail that forms the dense filament backbone. Myosin alone polymerizes into filaments of irregular length, but striated muscle thick filaments have defined lengths that with the thin filament define the sarcomere structure. The motor domain structure and function are well understood but the myosin filament backbone is not. Here we report a structure of the flight muscle thick filaments from Drosophila melanogaster at 4.7 Å resolution which eliminates previous ambiguities in the non-myosin densities. The full proximal S2 region is resolved as are the connecting densities between the Ig domains of stretchin-klp. The proteins, flightin and myofilin are resolved in sufficient detail to build an atomic model based on an AlphaFold prediction. Our results suggest how flightin and myofilin cooperate to define the structure of the thick filament and explains a key myosin mutation that affects flightin incorporation. Drosophila is a genetic model organism for which our results can define strategies for functional testing.

Environmental nanoparticles have been shown to exist in most aquatic environments, and environmental nanoparticles in water distribution systems significantly affect water quality. However, little research has been conducted on the environmental nanoparticles in mineral water, which is a special water resource. Nanoparticle tracking analysis (NTA) and high-resolution transmission electron microscopy (HRTEM) were conducted. This study was the first to identify many environmental nanoparticles in natural mineral water in Zibo City, Shandong Province, China. The results of the NTA showed that the concentration of the particles was 5.5 ×106 particles/mL and the peak diameter of the size distribution was 180 nm. The HRTEM showed that the nanoparticles were granular, pinniform, rodlike, and flakey in shape, and some of the nanoparticles existed in aggregation. The energy-dispersive spectrometry results showed that most of the nanoparticles contained O, Mg, Ca, Si, Fe, Ti, and P, and some of them also contained F, V, S, and Mn. When combined with the characteristics of the selected area electron diffraction pattern, the nanoparticles were confirmed to be Ca-bearing nanoparticles, attapulgite nanorods, MnO2 nanosheets, and TiO2 nanoparticles. These results indicate a new form of elements occurring in mineral water. In addition, based on the chemical and physical characteristics of the nanoparticles and mineral water, these environmental nanoparticles in the mineral water are likely to be the weathering byproducts of minerals. These nanoparticles within mineral water provide a good opportunity for understanding the behavior of nanoparticles in varied systems. Moreover, environmental nanoparticle science is important for continuing efforts in water safety, treatment, and remediation.

Over the past two decades, covalent organic frameworks (COFs) have become the most widely studied porous crystalline materials. Its specific physical and chemical properties are determined by the arrangement of atoms (crystal structure). Therefore, the determination of its structure is arguably the most important characterization step for COFs. Although single crystal X-ray diffraction (SCXRD) is the most widely used method for structure determination, the size of COF crystals is too small, or their crystal quality is so poor that confirmation of their structure is limited to lattice fringes by transmission electron microscopy (TEM). At present, many 2D-COFs have clear PXRD patterns, but not all 2D-COFs can obtain specific lattice fringes. This severely hinders the development of the COF field. Here, we discover the lattice shrinkage behavior of COFs under electron beam irradiation by comparing the lattice fringes of 2D-COFs under different conditions. By comparing the lattice fringes of TAPT-TFPT COF at room temperature and under liquid nitrogen freezing conditions, we found that the lattice fringes are in good agreement with the Powder X-ray diffraction (PXRD) and theoretical values of COF (2.213 nm) under freezing conditions, however, the lattice fringe spacing is only 1.656 nm at room temperature. The discovery not only provides new insights into the TEM characterization of COFs, but also further expands the range of crystalline COF materials.

The central nervous system (CNS) is an essential hub for neuronal communication. As a major component of the CNS, glial cells are vital in the maintenance and regulation of neuronal network dynamics. Research on microglia, the resident innate immune cells of the CNS, has advanced considerably in recent years, and our understanding of their diverse functions continues to grow. Microglia play critical roles in the formation and regulation of neuronal synapses, myelination, responses to injury, neurogenesis, inflammation and many other physiological processes. In parallel with advances in microglial biology, cutting-edge techniques for the characterization of microglial properties have emerged with increasing depth and precision. Labeling tools and reporter models are important for the study of microglial morphology, ultrastructure and dynamics, but also for microglial isolation, which is required to glean key phenotypic information through single-cell transcriptomics and other emerging approaches. Strategies for selective microglial depletion and modulation can provide novel insights into microglia-targeted treatment strategies in models of neuropsychiatric and neurodegenerative conditions, cancer and autoimmunity. Finally, fate mapping has emerged as an important tool to answer fundamental questions about microglial biology, including their origin, migration and proliferation throughout the lifetime of an organism. This review aims to provide a comprehensive discussion of these emerging techniques, with applications to the study of microglia in development, homeostasis and CNS pathologies.

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.

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.

The inhibition of localised corrosion is known to involve multiple processes taking place over a range of time and length scales that are difficult to study by conventional electrochemical methods. This work demonstrates an approach to probing complex localised electrochemical processes by combining an electrochemically integrated multi-electrode array (also known as wire beam electrode (WBE)), scanning vibrating electrode technique, scanning electrochemical microscopy and surface analytical techniques. Each technique reveals certain aspects of the dynamically changing, multi-scale, inhibitor-microstructure interactions and local chemistry over a heterogeneous AA2024-T3 alloy surface in the presence of an environmentally friendly inhibitor; cerium diphenyl phosphate (Ce(dpp)3).

Cellular biomolecular condensates termed ribonucleoprotein (RNP) granules often are enriched in messenger RNA (mRNA) molecules relative to the surrounding cytoplasm. Yet, the spatial localization and diffusion of mRNAs in close proximity to phase separated RNP granules is not well understood. In this study, we performed single molecule fluorescence imaging experiments of mRNAs in live cells in the presence of two types of RNP granules, stress granules (SG) and processing bodies (PB), which are distinct in their molecular composition and function. We developed a photobleaching- and noise-corrected colocalization imaging algorithm that was employed to determine the accurate positions of individual mRNAs relative to the granule’s boundaries. We found that mRNAs are often localized at granule boundaries, an observation consistent with recently published data. We suggest that mRNA molecules become spontaneously confined at the RNP granule boundary similar to the adsorption of polymer molecules at liquid-liquid interfaces, which is observed in various technological and biological processes. We also suggest that this confinement could be due to a combination of intermolecular interactions associated with, first, the screening of a portion of the RNP granule interface by the polymer and, second, electrostatic interactions due to a strong electric field induced by a Donnan potential generated across the thin interface.

Infectious keratitis (IK) is among the top 5 leading causes of blindness globally. Early diagnosis is key to guide an appropriate therapy to avoid complications such as vision impairment and blindness. Culture of corneal scrapes is the initial diagnostic test to grow and identify the causal organism. Alternative diagnostic tools include imaging such as with optical coherence tomography (OCT) and in vivo confocal microscopy (IVCM). OCT’s advantage is its ability to accurately determine the depth and extent of the corneal ulceration, infiltrates and haze; therefore, characterizing the severity and progression of the infection. However, it is not a preferred choice in the diagnostic tool package for infectious keratitis. IVCM is a great aid in the diagnosis of fungal and Acanthamoeba keratitis with overall sensitivities of 66-74% and 80-100%, and specificity of 78-100% and 84-100%, respectively. Recently the use of deep learning (DL) models in IK has been shown to aid diagnosis via image recognition. Most of the studies that have developed DL models to diagnose the different types of IK have utilised photographs from digital camaras, slit-lamp images, or IVCM images. Some studies have used extremely efficient single DL algorithms to train their models and others used ensemble approaches. This technology is likely to assist in the diagnosis of IK in some years. Further work is needed to examine and validate the clinical performance of the DL models in the real-world setting and to evaluate whether this technology improves patient clinical outcomes. The scope of this review was to provide a recent update on the diagnostic imaging tools in IK

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) often causes severe viral pneumonia, especially among older individuals and those with underlying health conditions. The C57BL/6 mouse is widely used to develop mouse models of obesity, diabetes, hypertension, and immune disorders, conditions that are associated with increased risk of severe COVID-19. Although many studies using these mouse models have examined the pathogenicity of SARS-CoV-2, COVID-19 pathogenesis remains poorly understood. In vivo imaging analysis using two-photon excitation microscopy is useful for elucidating the pathology of COVID-19, providing pathological insights that are not available from conventional histological analysis. However, there is no reporter SARS-CoV-2 that demonstrates pathogenicity in C57BL/6 mice and emits sufficient light intensity for two-photon in vivo imaging. Here, we generated SARS-CoV-2 expressing the fluorescent protein Venus to facilitate in vivo pathological studies using C57BL/6 mouse models. We generated a mouse-adapted strain of SARS-CoV-2, by performing sequential lung-to-lung passages in BALB/c mice, followed by in C57BL/6 mice. We observed that the C57BL/6-adapted SARS-CoV-2 (named MASCV2-p25) replicated efficiently in the lungs of C57BL/6 mice, causing fatal pneumonia. Histopathologic analysis revealed severe inflammation and infiltration of immune cells in the lungs of MASCV2-p25-infected C57BL/6J mice, not unlike that observed in COVID-19 patients with severe pneumonia. Subsequently, we generated a mouse-adapted reporter SARS-CoV-2 (named MASCV-Venus-p9) by inserting the fluorescent gene Venus into MASCV2-p25, and sequential lung-to-lung passages in C57BL/6 mice. C57BL/6 mice infected with MASCV2-Venus-p9 exhibited severe pneumonia similar to that induced by MASCV2-p25. In addition, two-photon excitation microscopy of the lungs of the infected C57BL/6J mice showed that the infected cells emitted sufficient levels of fluorescence for easy observation. These findings suggest that MASCV2-Venus-p9 will be useful for two photon in vivo imaging studies of the pathogenesis of severe COVID-19 and for the development of therapeutic agents for severe pneumonia.

Copper-nitride (Cu3N) material is attracting much attention as a potential next-generation thin-film solar-light absorber. In this work, polycrystalline Cu3N thin films were prepared using reactive-RF-magnetron sputtering, at room temperature,onto glass and silicon substrates. The optical properties of the Cu3N thin layers were studied by spectroscopic ellipsometry and transmission, to achieve a low-cost absorber material to replace conventional silicon. The reactive-RF-sputtered Cu3N films were also investigated by FIB-SEM microscopy and FTIR spectroscopy.

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 term neuroinflammation defines the reactions of astrocytes and microglia to alterations in homeostasis in the diseased central nervous system (CNS), the exacerbation of which contributes to the neurodegenerative effects of Alzheimer's disease (AD). Local environmental conditions, such as the presence of proinflammatory molecules, mechanical properties of the extracellular matrix (ECM) and local cell-cell interactions, are determinants of glial cell phenotypes. In AD, the load of the cytotoxic/proinflammatory amyloid β peptide (Aβ) is a microenvironmental component increasingly growing in the CNS, imposing time-evolving challenges on resident cells. This study aimed at investigating the temporal and spatial variations of the effects produced by this process on astrocytes and microglia, either directly or by interfering in their interactions. Ex vivo confocal analyses of hippocampal sections from the mouse model TgCRND8 at different ages have shown that overproduction of Aβ peptide induced early and time persistent disassembly of functional astroglia syncytium and promoted a senile phenotype of reactive microglia, hindering Aβ clearance. In the late stages of disease, these patterns were altered in the presence of Aβ-plaques, surrounded by typically reactive astrocytes and microglia. Morphofunctional characterization of peri-plaque gliosis revealed a direct contribution of astrocytes in plaque buildup that might result in shielding Aβ-peptide cytotoxicity and, as a side effect, in exacerbating neuroinflammation.

Macromolecular assemblies, such as protein complexes, undergo continuous structural dynamics, including global reconfigurations critical for their function. Two fast analytical methods are widely used to study these global dynamics, namely elastic network model normal mode analysis and principal component analysis of ensembles of structures. These approaches have found wide use in various computational studies, driving the development of complex pipelines in several software packages. One common one has been conformational sampling through hybrid simulations incorporating all-atom molecular dynamics and global modes of motion. However, wide functionality is only available for experienced programmers with limited capabilities for other users. We have, therefore, integrated one popular and extensively developed software for such analyses, the ProDy Python application programming interface, into the Scipion workflow engine. This enables a wider range of users to access a complete range of macromolecular dynamics pipelines beyond the core functionalities available in its command-line applications and the normal mode wizard in VMD. The new protocols and pipelines can be further expanded and integrated into larger workflows together with other software packages for cryo-electron microscopy image analysis and molecular simulations. We present the resulting plugin, Scipion-EM-ProDy, in detail, highlighting the rich functionality made available by its development.

Migratory flows and international travel are triggering an increase in imported cases of schistosomiasis in non-endemic countries, with serology being the recommended method for screening migrants. The present study aims to evaluate, for the first time, the LAMP technique on patients’ urine samples for the diagnosis of imported schistosomiasis in a non-endemic area in comparison to a commercial immunochromatographic test and microscopic examination of feces and urine. A prospective observational study was conducted in sub-Saharan migrants attending at the Tropical Medicine Unit, Almería, Spain. For schistosomiasis diagnosis, serum samples were tested using an immunochromatographic test (Schistosoma ICT IgG-IgM). Stool and urine samples were examined by microcopy. Urine samples were evaluated by combining three LAMP assays for the specific detection of Schistosoma mansoni and S. haematobium, and an assay for the genus Schistosoma. To evaluate the diagnostic accuracy of Schistosoma-LAMP in comparison to microscopy and Schistosoma ICT IgG-IgM test, a Latent Class Analysis (LCA) was performed. A total of 115 patients were included; 21 patients (18.3%) were diagnosed with schistosomiasis confirmed by microscopy, with S. haematobium being the most frequent species identified (18/115; 15.7%). The Schistosoma ICT IgG-IgM test resulted 100% positive and Schistosoma-LAMP was 61.9% positive, reaching as high as 72.2% for S. haematobium. The sensitivity and specificity estimated by LCA, respectively, were: 92% and 76% for Schistosoma ICT IgG-IgM, 68% and 44% for Schistosoma-LAMP and, 46% and 97% for microscopy. The Schistosoma-LAMP technique presented a higher sensitivity than microscopy for the diagnosis of imported urinary schistosomiasis, which could improve the diagnosis both in referral centers and in centers with limited experience or scarce resources and infrastructure.

The morphology of virgin reactor powder (RP) of high-density polyethylene HDPE with MW = 160,000 g/mol was investigated with the help of DSC, SEM, SAXS and WAXS methods. The morphological SEM analysis showed, that the main morphological units of RP are macro- and micro-shish kebab structures with significantly different geometric dimensions, as well as individual lamellae of folded chain crystals. Quantitative analysis of an asymmetric SAXS reflection made it possible to reveal the presence of several periodic morphoses in the RP with long periods from 20 nm to 60 nm and to correlate them with the observed powder morphology. According to the DSC crystallinity data, the thickness of the lamellae in each long period was estimated. Their surface energy was calculated in the framework of the Gibbs-Thompson theory. The presence of regular and irregular folds on the surface of the different shish-kebab lamellae is discussed. The percentage of identified morphoses in the RP was calculated. It has been suggested that the specific structure of HDPE RP is due to the peculiarity of polymer crystallization during suspension synthesis in a quasi-stationary regime, in which local overheating and inhomogeneous distribution of shear stresses in a chemical reactor are possible.

Many studies of hypervelocity impact craters have described the characteristics of quartz grains shock-metamorphosed at high pressures of > 10 GPa. In contrast, few studies have investigated shock metamorphism at lower shock pressures. In this study, we test the hypothesis that low-pressure shock metamorphism occurs in near-surface nuclear airbursts and that this process shares essential characteristics with crater-forming impact events. To investigate low-grade shock microstructures, we compared quartz grains from Meteor Crater, a 1.2-km-wide impact crater, to those from near-surface nuclear airbursts at the Alamogordo Bombing Range, New Mexico in 1945 and Kazakhstan in 1949/1953. This investigation utilized a comprehensive analytical suite of high-resolution techniques, including transmission electron microscopy (TEM) and electron backscatter diffraction (EBSD). Meteor Crater and the nuclear test sites all exhibit quartz grains with closely-spaced, sub-micron-wide fractures that appear to have formed at low shock pressures. Significantly, these micro-fractures are closely associated with Dauphiné twins and are filled with amorphous silica (glass), widely considered a classic indicator of shock metamorphism. Thus, this study confirms that glass-filled shock fractures in quartz form during near-surface nuclear airbursts, as well as crater-forming impact events, and by extension, it suggests that they may form in any near-surface cosmic airbursts in which the shockwave is coupled to Earth’s surface. The robust characterization of such events is crucial because of their potential catastrophic effects on the Earth’s environmental and biotic systems.

Coded aperture 3D imaging techniques have been rapidly evolving in the recent years. The two main directions of evolution are in aperture engineering to generate the optimal optical field and in development of computational reconstruction to reconstruct the object’s image from the intensity distribution with a minimal noise. The goal is to find the ideal aperture-reconstruction method pair and if not, to optimize one to match the other for designing an imaging system with required 3D imaging characteristics. Lucy-Richardson-Rosen algorithm (LR²A), a recently developed computational reconstruction method was found to perform better than its predecessors such as matched filter, Weiner filter, phase-only filter, Lucy-Richardson algorithm and non-linear reconstruction (NLR) for certain apertures when the point spread function (PSF) is a real and symmetric function. For other cases of PSF, NLR performed better than the rest of the methods. In this tutorial, LR²A has been presented as a generalized approach for any optical field along with MATLAB codes for reconstruction of any image when the PSF is known. The common problems and pitfalls in using LR²A has been discussed. Simulation and experimental studies for common optical fields such as spherical, Bessel, vortex beams and exotic optical fields such as Airy, scattered and self-rotating beams have been presented. From this study, it can be seen that it is possible to transfer the 3D imaging characteristics from non-imaging type exotic fields to indirect imaging systems faithfully using LR²A. The application of LR²A to medical images such as colonoscopy images and cone beam computed tomography images with synthetic PSF has been demonstrated. We believe that the tutorial will provide a deeper understanding of computational reconstruction using LR²A.

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.

Under inflammatory conditions including lymphatic disorders, bone marrow-derived myeloid cells often express lymphatic endothelial cell (LEC) markers called LEC progenitor cells, which extend lymphatic vessels by fusing with existing lymphatic vessels. However, studies on the mechanism of lymphatic regeneration using three-dimensional images of lymphatic structures are limited. In this study, scanning electron microscopy (SEM) was used to observe the three-dimensional structure of lymphangiogenesis in a mouse model of secondary lymphedema. The model was established in C57BL/6J mice via circumferential incision in the inguinal region of the left hind limb. Skin samples were obtained from the lymphedema region on days 2, 5, and 8 after surgery. To determine lymphatic vessel positions using SEM analysis, we detected anti-lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1) immunoreactivity in serial sections and overlaid them during SEM observation. On days 2 and 5, spherical cells, probably myeloid cells, were attached and fused to the LYVE-1-positive lymphatic vessel walls. On day 8, spherical cells were converted to string-shaped cells, forming a new lymphatic vessel wall resembling an intraluminal pillar. Our results show the three-dimensional lymphatic structures formed during lymphatic regeneration.

(1) Background: Stereological estimations significantly contributed to our anatomical and physiological understanding of the lung by providing an average distribution of measured parameters over the entire lung parenchyma. However, most structural lung diseases show a very inhomogeneous pattern of alterations. (2) Methods: We developed a novel protocol for the analysis of all pulmonary airspaces. Our pipeline starts with high-resolution synchrotron radiation-based X-ray tomographic microscopy (SRXTM) and consists of (i) image segmentation with the combination of the free machine-learning tool Ilastik and ImageJ and (ii) calculation of the airspace diameter distribution of an entire lung using a diameter map function. To evaluate the new pipeline, lungs from adult mice with cystic fibrosis (CF)-like lung disease (ENaC-transgenic mice) or mice with elastase-induced emphysema were compared to healthy controls. (3) Results: We were able to show the distribution of airspace diameters throughout the entire lung, as well as separately for the conducting airways and the gas-exchange area. In the pathobiological context, we observed an irregular widening of parenchymal airspaces in mice with CF-like lung disease and elastase-induced emphysema. Comparable results were obtained when analyzing lungs imaged with μCT, suggesting that our pipeline is applicable to different kinds of imaging modalities. (4) Conclusions: We conclude that the airspace diameter map is well suited for a detailed analysis of unevenly distributed structural alterations in chronic muco-obstructive lung diseases such as cystic fibrosis and COPD.

As sessile organisms, plants face a wide range of abiotic stresses, with salinity being a prominent constraint affecting their growth, development, and productivity, particularly in arid and semi-arid regions. This study focused on understanding how salinity impacts Jatropha curcas, an important oilseed plant for biodiesel production. By examining the anatomy and ultrastructure of stomata and chloroplasts, we investigated the effects of prolonged salinity stress on J. curcas. This stress led to changes in stomatal density, stomatal index, and ostiole aperture, which case an imbalance of water conductivity in the xylem. Through transmission electron microscopy, we explored the subcellular organization of J. curcas chloroplasts and their contribution to plant photosynthetic efficiency, providing insights into their role in this process. Notably, salinity treatment resulted in a significant increase in starch granules accumulation, leading to impaired the granal and stromal grana lamellae, destroying this ultrastructure. Our findings indicate that the anatomy and ultrastructure of chloroplasts play a crucial role in influencing photosynthetic efficiency and hydraulic conductivity. This study offers new perspectives on the structure and function of chloroplasts in J. curcas, presenting innovative opportunities to develop strategies that enhance biofuel production in areas with high soil salinity.

Is there any possible synergic effect of fertility outcome between spindle view-assisted intracyto-plasmic sperm injection (SV-ICSI) with or without assisted oocyte activation (AOA) for patients with low fertilization rate history? To explore the answer, a retrospective study was held in our fertility center by reviewing patients receiving SV-ICSI during 2019/03 to 2023/06. Total 47 pa-tients were included, the fertility outcomes of intervention group (AOA-SV-ICSI, 14 patients) and control group (SV-ICSI, 33 patients), such as fertilization rate, blastocyst formation rate and clin-ical pregnancy rate, were analyzed. The blastocyst formation rate between groups was signifi-cantly higher in AOA-SV-ICSI group (p=0.020), but the fertilization rate and clinical pregnancy rate did not reach any statistical significance. This discovery of increased blastocyst formation rate in low fertilization rate patients could possibly help them in getting more usable embryos in fertility treatment.

Djeya1 (RKLAFRYRRIKELYNSYR), is a very effective cell penetrating peptide (CPP) which mimics the α5 helix of the highly conserved Eya domain (ED) of eyes absent (Eya) proteins. The objective of this study was to bioengineer analogues of Djeya1 that, following effective translocation into planarian tissues, would reduce the ability of neoblasts (totipotent stem cells) and their progeny to regenerate the anterior pole in decapitated S. mediterranea. As a strategy to increase the propensity for helix formation, molecular bioengineering of Djeya1 was achieved by the mono-substitution of the helicogenic aminoisobutyric acid (Aib) at three species variable sites: 10,13,16. CD analyses indicted that Djeya1 is highly helical and that Aib-substitution had little influence on secondary structure. Aib-substituted Djeya1 analogues are highly efficient CPPs, devoid of influence upon cell viability or proliferation. All three peptides increase the migration of PC-3 cells which express high concentrations of Eya. Two peptides, [Aib13]Djeya1 and [Aib16]Djeya1, are bioportides which delay planarian head regeneration. As neoblast are the only cell population capable of division in planaria, these data indicate that bioportide technologies could be utilised to directly manipulate other stem cells in situ, thus negating any requirement for genetic manipulation.

Magnetic and morphological properties of equiatomic B2-ordered FeRh thin films irradiated with single high-intensity ultrashort laser pulses are investigated. The goal is to elucidate the effect of femtosecond laser ablation on the magnetic properties of FeRh. We employed Scanning Magneto-Optical Kerr Effect (S-MOKE) microscopy to examine the magnetic phase after laser processing, providing high spatial resolution and sensitivity. Our results revealed the appearance of a magneto-optical signal from the bottom of ablation craters, suggesting a transition from antiferromagnetic to ferromagnetic behavior. Fluence-resolved measurements clearly demonstrate that the ablation threshold coincides with the threshold of the antiferromagnet-to-ferromagnet phase transition. The existence of such magnetic phase transition was independently confirmed by temperature-dependent S-MOKE measurements using a CW laser as a localized heat source. Whereas the initial FeRh film displayed a reversible antiferromagnet-ferromagnet phase transition, the laser-ablated structures exhibited irreversible changes in their magnetic properties. This comprehensive analysis revealed the strong correlation between the femtosecond laser ablation process and the magnetic phase transformation in FeRh thin films.

Lichens are unique extremophilic organisms due to their phenomenal resistance to adverse environmental factors, including ultraviolet (UV) irradiation. Melanization plays a special role in the protection of lichens from UV-B stress. In present study, we analyzed the binding of melanins with the components of cell walls of mycobiont of upper cortex in the melanized lichen thalli Lobaria pulmonaria. Using scanning electron and atomic force microscopy, the morphological and nanomechanical characteristics of melanized layer of mycobiont cells were visualized. Melanization of lichen thalli led to the smoothing the surface relief and thickening mycobiont cell walls and resulted in the increase of deformation values and reduction of adhesion properties of lichen thallus. Treatment of thalli with hydrolytic enzymes, especially chitinase and lichenase, enhanced the yield of melanin from melanized thalli and release of carbohydrates, while treatment with pectinase increased the release of carbohydrates and phenols. Our results suggest that melanin by firmly binding with hyphal cell wall carbohydrates, particularly chitin and 1,4-β-glucans, can form associations, strengthen the melanized upper cortex of lichen thalli, and thereby can contribute to lichen survival under UV stress.

Unlike plants which have special gravity-sensing cells, such special cells in animals are yet to be discovered. However, microgravity, the condition of apparent weightlessness, causes bone, muscular and immune system dysfunctions in astronauts following spaceflights. Decades of investigations show correlations between these organ and system-level dysfunctions with changes induced at the cellular level both by simulated microgravity as well as microgravity conditions in outer space. Changes in single bone, muscle and immune cells include morphological abnormalities, altered gene expression, protein expression, metabolic pathways and signaling pathways. These suggest that human cells mount some response to microgravity. However, the implications of such adjustments on many cellular functions and responses are not clear. Here, we addressed the question whether microgravity induces alterations to drug response in cancer cells. We used both adherent cancer cells (T98G) and cancer cells in suspension (K562) to confirm known effects of microgravity and then treated the K562 cells with common cancer drugs (hydroxyurea and paclitaxel) following 48 hours of exposure to microgravity via a NASA-developed rotary cell culture system. Fluorescence-guided morphometry revealed microgravity-induced loss of the significant reduction (p < 0.0l) to the nuclear to cytoplasm ratio of cancer cells treated with hydroxyurea. Our results call for more studies on impact of microgravity on cellular drug-response, in view of the growing need for space medicine, as space exploration grows.

When oxidizing reduced sulfur compounds, the phototrophic sulfur bacterium Allochromatium vinosum forms spectacular sulfur globules as obligatory intracellular – but extracytoplasmic – intermediates. The globule envelope consists of three extremely hydrophobic proteins: SgpA and SgpB are very similar and can functionally replace each other. SgpC is involved in the expansion of the sulfur globules. The presence of a fourth protein, SgpD, has been suggested by comparative transcriptomics and proteomics of enriched sulfur globules. Here, we investigated the in vivo function of SgpD by coupling its carboxy-terminus to mCherry. This fluorescent protein requires oxygen for chromophore maturation, but we were able to use it in anaerobically growing A. vinosum provided the cells were exposed to oxygen for one hour prior to imaging. While mCherry lacking a signal peptide resulted in low fluorescence evenly distributed throughout the cell, fusion with SgpD carrying its original Sec-dependent signal peptide targeted mCherry to the periplasm and co-localized it exactly with the highly light-refractive sulfur deposits seen in sulfide-fed A. vinosum cells. Insertional inactivation of the sgpD gene showed that the protein is not essential for the formation and degradation of sulfur globules.

Urinary tract infections are among the most frequent infectious diseases and require screening of a great amount of urine samples from patients. However, a high percentage of samples results as negative after urine culture plate test (CPT), demanding a simple and fast preliminary technique to screen out the negative samples. We propose a digital holographic microscopy (DHM) method to inspect fresh urine samples flowing in a glass capillary for 3 minutes, recording holograms at 2 frames per second. After digital reconstruction, bacteria, white and red blood cells, epithelial cells and crystals were identified and counted and the samples were classified as negative or positive according to clinical cutoff values. Taking CPT as reference, we processed 180 urine samples and compared the results with those of urine flow cytometry (UFC). Using standard evaluation metrics for our screening test, we found similar performance for DHM and UFC, indicating DHM as a suitable and fast screening technique retaining several advantages. As a benefit of DHM, the technique is label-free and does not require sample preparation. Moreover, the phase and amplitude images of the cells and other particles present in urine are digitally recorded and can serve for further investigation afterwards.

The purpose of this study was to describe ocular surface and anterior eye segment findings in various types of ichthyoses. Methods: This was a single-center prospective observational study. The study group consisted of five patients (P1-P5) aged 13–66 years. Multimodal imaging was performed, including slit-lamp examination, swept-source optical coherence tomography (SS-OCT), and in vivo confocal microscopy (IVCM). Results: All patients were diagnosed with moderate to severe dry eye disease (DED). Corneas showed a significant pattern of irregularity, with a significant difference between the corneal thickness at the apex (CAT) and corneal thinnest thickness (CTT), ranging from 5 to 375 µm. Three patients were diagnosed with ectasia patterns based on SS-OCT. All patients showed abnormalities in at least one Fourier index parameter for at least one eye at 3 or 6 mm, on keratometric, anterior, or posterior analyses. IVCM examination revealed changes in all corneal layers. Conclusions: By combining the results of multimodal im-aging, we were able to detect preclinical abnormalities, distinguish characteristic changes com-mon to ichthyosis, and reveal the depth and characteristics of corneal abnormalities. Therefore, patients with ichthyosis should be examined for DED and ectatic disorders early in clinical prac-tice.

Glycerol has found its applications as a heat-transfer fluid in heat exchangers, and as a compo-nent of functional solutions in biosensor analysis. Flowing non-aqueous fluids are known to be able to induce electromagnetic fields due to triboelectric effect. These triboelectrically generated electromagnetic fields can affect biological macromolecules. Horseradish peroxidase (HRP) is widely employed as a convenient model object for studying how external electric, magnetic, and electromagnetic fields affect enzymes. Herein, we have studied whether the flow of glycerol in a ground-shielded cylindrical coil affects the HRP enzyme incubated at a 2 cm distance near the coil’s side. Atomic force microscopy (AFM) has been employed in order to study the effect of glycerol flow on HRP at the nanoscale. Increased aggregation of HRP on mica has been observed after the incubation of the enzyme near the coil. Moreover, the enzymatic activity of HRP has also been affected. The results reported can find their application in biotechnology, food technology and life sciences application, considering the development of triboelectric generators, en-zyme-based biosensors and bioreactors with surface-immobilized enzymes. Our data can also be of interest for scientists studying triboelectric phenomena.

The phenomenon of pathogens co-infection detected in the half fed on humans I. persulcatus tick in the south of the Far East was studied. Researchs were carried out on PEK, Vero, Vero-E6 cell lines, outbred mice, chicken embryos, using ELISA, PCR, IMFA, plaque formation, and electron microscopy. The tick contained an antigen and a genetic marker of the tick-borne encephalitis virus (TBEV). The patient had post-vaccination antibodies in a titer of 1:200, as a result of which, obviously, an antibody-dependent elimination of TBEV occurred. The tick-borne co-isolate also contained an unknown pathogen (Kiparisovo-144 virus), which, in our opinion, was a trigger for the activation of chronic infection in suckling white mice. In the laboratory coisolate, ectromelia virus was present, as evidenced by paw edema during intradermal infection of mice, characteristic rashes on the chorion-allantoic envilope of chicken embryos, and typical plaques on Vero-E6. The Kiparisovo-144 virus was not pathogenic for white mice and chicken embryos, but successfully multiplied in the PEK, Vero, and Vero-E6 lines. Viral co-infection was confirmed by electron microscopy. Passaging on mice contributed to an increase in the virulence of the co-isolate, whose titer increased by 10,000 times by the 5th passage, which poses a serious epidemiological danger.

We have investigated the interfacial dislocations in InxGa1–xN/GaN (0 ≤ x ≤ 0.20) heterostructures using diffraction contrast analysis in a transmission electron microscopy. The analysis indicate that the structural properties of the interface dislocations depend on the indium composition. For lower indium composition up to x = 0.09, we observed that the screw-type dislocations and dislocation half loops occurred at the interface even though the former do not contributes toward elastic relaxation of the misfit strain in the InGaN layer. With the increase of indium composition (0.13 ≤ x ≤ 0.17), in addition to the network of screw-type dislocations, edge-type misfit dislocations were found generated with their density gradually increasing. For higher indium composition (0.18 ≤ x ≤ 0.20), all the interfacial dislocations are transformed into a network of straight misfit dislocations along the <10-10> directions leading to partially relaxation of the InGaN epilayer. The presence of dislocation half loops may be explained by slip on the basal plane, the formation of misfit dislocations are attributed to punch-out mechanism.

Many studies of hypervelocity impact craters have described the characteristics of quartz grains shock-metamorphosed at high pressures of >10 GPa, but in contrast, few studies have investigated shock metamorphism at lower shock pressures. In this study, we test the hypothesis that low-pressure shock metamorphism occurs in near-surface nuclear airbursts and that this process shares important characteristics with impact-cratering events. To investigate low-grade shock microstructures, we compared quartz grains from Meteor Crater, a 1.2-km-wide impact crater, to those from near-surface nuclear airbursts at the Alamogordo Bombing Range, New Mexico in 1945 and Kazakhstan in 1949/1953. This investigation utilized a comprehensive analytical suite of high-resolution techniques, including transmission electron microscopy (TEM) and electron backscatter diffraction (EBSD). Meteor Crater and the nuclear test sites all exhibit metamorphosed quartz grains with closely-spaced, sub-micron-wide fractures that appear to have formed at low shock pressures. Importantly, these micro-fractures are closely associated with Dauphiné twins and are filled with amorphous silica (glass), widely considered to be a classic indicator of shock metamorphism. Thus, this study confirms that glass-filled shock fractures in quartz form during near-surface nuclear airbursts, as well as crater-forming impact events, and by extension, it suggests they also may form in near-surface cosmic airbursts.

Dawn of the omics revolution in biological sciences meant that we seek to understand more and in greater detail the molecular constituents of cells and biological systems. While we have gained significant insights from conventional omics tools, we now seek to understand the spatial dimensions of the data where subcellular localisation may impact on cellular physiology and phenotype. This review paper seeks to address current questions in the new field of image-based spatial proteomics as well as outline future challenges of the field. At first glance, spatial proteomics offers enormous potential to expand our understanding of different cell types in different disease and cell states. But limitations in types of fluorophores and issues with spectral overlap significantly hampers the practical implementation of the technique. On the other hand, while we have super-resolution microscopy techniques such as STED, PALM and STORM able to achieve 10 to 20 nm spatial resolution in single molecule localisation, problems with slow image acquisition limits high temporal resolution tracking of multiple protein targets in live cell imaging. Hence, the field of spatial proteomics is a mix of promises and challenges where we could image, in multi-colour, upwards of 10 well-chosen proteins that could inform on the molecular mechanisms of selected biological processes, but, at present, the method could not tackle larger scale questions. In essence, current implementation of image-based spatial proteomics is useful, but it is unable to fulfil the mission of large-scale projects such as the Human Protein Atlas or Human Cell Atlas. Future challenges in the field includes the development of more fluorophores (especially photoswitchable and photoactivable ones) for single molecule localisation microscopy, as well as seeking to improve temporal resolutions to the sub-millisecond range.

Common bean has received attention as a model plant for legume studies, but little information is available about the morphology of its pods and the relation of this morphology to loss of seed dispersal and/or the pod string, which are key agronomic traits of legume domestication. Dehiscence is related to pod morphology and the anatomy of pod tissues because of the weakening of the dorsal and ventral dehiscence zones and the tensions of the pod walls. These tensions are produced by the differential mechanical properties of lignified and non-lignified tissues and changes in turgor associated with fruit maturation. In this research, we histologically studied the dehiscence zone of the ventral and dorsal sutures of the pod in two contrasting genotypes for dehiscence and string, by comparing different histochemical methods with autofluorescence. We found that the secondary cell wall modifications of the ventral suture of the pod were clearly different between the dehiscence-susceptible and stringy PHA1037 and the dehiscence-resistant and stringless PHA0595 genotypes. The susceptible genotype had cells of bundle caps arranged in a more easily breakable bowtie knot shape. The resistant genotype had a larger vascular bundle area and fibre cap cells (FCCs), and due to their thickness, the external valve margin cells were significantly stronger than those from PHA1037. Our findings suggest that the FCC area, and the cell arrangement in the bundle cap, might be a partial structure involved in the pod dehiscence of common bean. The autofluorescence pattern at the ventral suture allowed us to quickly identify the dehiscent phenotype and gain a better understanding of cell wall tissue modifications that took place along the bean's evolution, which had an impact on crop improvement. We report a simple autofluorescence protocol to reliably identify secondary cell wall organization and its relationship to dehiscence and string in common bean.

Ni1-xGdxFe2O4 (x=0.02, 0.04, 0.06 & 0.08) (NGF) nanoparticles were synthesized via hydrothermal method. The X-ray diffraction study confirmed the single-phase cubic spinel structure. The morphology indicated the stones (approximately nanocubes) like structures in the FESEM images. The mass loss, and thermal transition temperatures were evaluated as a function of Gd-content using TG-DTG, and TG-DSC measurements. The M-H loops evidenced the high magnetization values for NGFO as compared to pure nickel ferrite. Cation distribution showed the preferred occupancy of cations in the magnetic spinel system. Frequency dependence of dielectric, conductivity, and impedance parameters was elucidated with increase in Gd-content.

Ribosome assembly is one of the most fundamental processes in gene expression and has served as a playground to investigate the molecular mechanisms of how protein-RNA complexes (RNPs) assemble. The bacterial ribosome is composed of around 50 ribosomal proteins several of which are co-transcriptionally assembled on a ~4,500 nucleotides long pre-rRNA transcript that is further processed and modified during transcription, the entire process taking around 2 minutes in vivo and assisted by dozens of assembly factors. How this complex molecular process works so efficiently to produce an active ribosome has been investigated over decades and has resulted in the development of a plethora of novel approaches that can also be used to study the assembly of other RNPs. Here we review biochemical, structural and biophysical methods that have been developed and integrated to provide a detailed and quantitative understanding of this complex and intricate molecular process of assembly. We also discuss emerging cutting-edge approaches that could be used in the future to study how transcription, rRNA processing, cellular factors and the native cellular environment shape ribosome assembly and RNP assembly at large.

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

The precise differentiation of renal cell tumours (RCTs) is sometimes hard to achieve using standard imaging and histopathological methods. This study investigated 43 cell renal cell car-cinomas (ccRCC), 15 papillary renal cell carcinomas (pRCC), 20 chromophobe renal cell carcino-mas (chRCC), and 18 renal oncocytomas (RO), stained with anti-mitochondria antibody (Thermo Scientific) by immunohistochemistry and immunofluorescence, and assessed by electron micros-copy, in order to define mitochondria distribution pattern (coarse scanty, moderate granular and diffuse granular). Thus, the majority of males had coarse granular staining in the tumours, while females were almost equally distributed among groups (p=0.005). An average patient age, tumour side and dimension, and tumour stage were similar in all staining pattern groups. However, pathohistological tumour types had significantly different expression patterns, with the lower amount of staining detected in majority of ccRCC, moderate expression in all chRCC, and diffuse expression in all RO, pRCC and two cases of ccRCC (p<0.001) presented with higher nuclear grade (p=0.005). Moreover, with increased distribution of mitochondria, the intensity of staining was higher (p<0.001). Here we present a strategy that utilizes mitochondria detection to differentiate RO from chRCC, as well as to distinguish other frequent RCTs, such as ccRCC and pRCC.

Metal-organic frameworks (MOFs), composed of metal nodes and inorganic linkers are promising for a wide range of applications due to their unique periodic frameworks that can be flexibly tuned. Understanding of the structure-activity relationships can facilitate the development and application of MOFs nanomaterials. Transmission electron microscopy (TEM) is a one-of-a-kind technique capable of characterizing MOFs microstructures at the atomic scale. Besides, in-situ TEM set-ups make it possible to direct visualize the microstructural evolution of MOFs in real time, which enables monitoring transitions of MOFs during growth as well as under working conditions. However, MOFs are so sensitive to the high-energy electron beam which makes TEM-based characterizing methods challenging. In this review, we first introduce the main damage mechanisms for MOFs under electron beam irradiation and two strategies to minimize the damages: low-dose TEM and cryo-TEM. Then, we discuss the techniques combined with the two strategies to analyze the MOFs’ microstructure, including three-dimensional electron diffraction, direct detection electron counting camera, and iDPC-STEM. Groundbreaking milestones and research advances with respect to static characterization of MOFs structures are highlighted. In-situ TEM studies are exemplary reviewed to provide insights into MOFs dynamics induced by various stimuli. Additionally, perspectives on promising techniques that may further facilitate the MOFs study using TEM are analyzed.

For the replacement of lost contractile elements in the treatment of coronary heart disease, the most promising approaches to obtain cardiomyocytes by cardiac differentiation of pluripotent cells. The aim of this work is to develop a technology for the formation of a functional layer of cardiomyocytes differentiated from iPSCs, capable of generating rhythmic activity and synchronous contractions. To accelerate cardiomyocyte maturation, the renal subcapsular transplantation model was used in SCID mice. After explantation, the formation of cardiomyocyte contractile apparatus was assessed by fluorescence and electron microscopy, and calcium ion oscillation in the cytoplasm was assessed by visualization of the fluorescent calcium binding dye Fluo-8. It was shown that human iPSC-derived cardiomyocyte cell layers transplanted (for up to 6 weeks) under SCID mouse kidney fibrous capsules begin to form an ordered contractile apparatus and retain functional activity and the ability to oscillate calcium ion fluxes after explantation from the body.

Imaging of the ocular vasculature can provide new insights into the pathophysiology of ocular diseases. This study proposes a novel high-frequency super-resolution ultrasound localization microscopy (SRULM) technique and evaluates its ability to measure in vivo perfusion changes in the rat eye at elevated intraocular pressure (IOP). A 38.4 MHz center frequency linear array transducer on a VisualSonics Vevo F2 imaging platform was used to collect high frame rate (1 kHz) radiofrequency data of the posterior rat eye following systemic microbubble contrast injection. Following clutter and spatiotemporal non-local means filtering, individual microbubbles were localized and tracked. The microbubble tracks were accumulated over 10,000 frames to generate vascular images quantifying perfusion velocity and direction. Experiments were performed using physiologic relevant controlled flow states for algorithm validation and subsequently performed in vivo on the rat eye at 10 mm Hg IOP increments from 10 to 60 mm Hg. The posterior vasculature of the rat eye, including the ophthalmic artery, long posterior ciliary arteries and their branches, central retinal artery and retinal arterioles and venules were successfully visualized, and velocities quantified at each IOP level. Significant reductions in arterial flow were measured as IOP was elevated. High-frequency SRULM can be used to visualize and quantify perfusion velocity of the rat eye in both the retrobulbar and intraocular vasculature simultaneously. The ability to detect ocular perfusion changes throughout the depth of the eye may help elucidate the role ischemia has in the pathophysiology of ocular diseases such as glaucoma.