To detect hydrogen in materials at the atomic scale, atom probe tomography is now regularly used. In order to avoid cumbersome cryo-preparation to suppress diffusion, often hydrogen is charged only into the finished specimen. Here, the use of hydrogen gas over electrochemical hydrogen has the advantage that the specimen is not contaminated with an electrolyte. So far, this ‘charging’ has been done in large, expensive systems. Here, we introduce small devices that enable the exposure of atom probe specimens to hydrogen and potentially other gases, using only very small gas volumes. This enables the operation in regular laboratory environments without additional safety measures. These devices can be used to expose the specimen to hydrogen up to 10 bar / 90 °C. Higher temperatures may be attained with small changes. Validation of the success of charging with these setups is demonstrated through experiments employing deuterium charging of palladium atom probe tips.

This study was carried out to investigate the macro-anatomical, morphometric, histological and electron microscopic characteristics of the uropygial gland in adult male and female turkeys (Meleagris Gallapova). The glandula uropygialis of thirteen adult turkeys (6 males, 7 females) were used for this study. It was determined that this glandular structure located dorsal to the last caudal vertebrae was heart-shaped and consisted of two lobes and a papilla system. As a result of statistical evaluation, significant differences were found between the sexes in the parameters of lobe length, papilla width and papilla height (P<0.05). Also, PUI value had a statistically significant difference (P<0.05). Tissue samples taken to determine the histological structure of the gland were stained with Haematoxylin & Eosin, Crossman’s and Periodic Acid Schiff. Histological examination revealed that the gland had a two-lobed structure surrounded by a capsule composed of connective tissue. It was determined that the gland had a tubuloalveolar-holocrine structure and the epithelial layer consisted of cellular layers as germinative layer, intermediate layer, secretory layer and degenerative layer from the periphery to the centre. In the scanning electron microscope examination, the lobe structure of the gland consisted of different shaped follicles protruding from the surface. It was observed that these glandular follicle structures were gathered together in different ways or mostly disorganised. As a result of the study, it was determined that the morphological and histological structure of the uropygial gland in turkeys was similar to that of other bird species, but showed some species-specific and habitat-dependent differences in general. Research Highlights: The uropygial gland consisted of a system of two lobes and one papilla in each of male and female turkeys. The glandular lobes consisted of follicles surrounded by connective tissue, while the follicles were connected to each other by interfollicular septae. The results obtained in morphometric measurements revealed statistical differences between male and female birds. Histological examination showed PAS (+) reaction in the basal membranes of tubule epithelial cells and secretion. Lymphoid cell communities were found in both connective tissue and intertubular regions.

Skeletal porous lesions such as cribra orbitalia (CO) have long been of interest to bioanthropologists worldwide, mainly due to their high prevalence in osteological material. Previous studies considered CO as an external morphological manifestation, and therefore, research has mainly focused on visible (macroscopic) CO patterns. However, the understanding of CO-induced micro-scale bone changes is still scarce. Therefore, here we performed high-resolution micro-computed tomography imaging to investigate three-dimensional CO-induced micro-architectural patterns in non-adults, with a particular focus on the correlation between macroscopic and micro-architectural orbital features. Cortical and trabecular micro-architectural changes in the orbital roof were analyzed in non-adults up to 15 years of age, using orbital roof samples with and without macroscopic traces of CO (n=28). A widely accepted five-grade macroscopic CO scoring system was applied to analyze CO severity. Areas affected with CO (area 1) and areas without macroscopic CO traces (area 2) were analyzed separately. The conducted high-resolution analysis showed that cortical and trabecular micro-architecture varied with CO presence, lesion severity (CO grade), and the analyzed area. Inter-grade comparisons suggested that most of the analyzed micro-architectural parameters were not significantly different between adjacent CO grades. Based on the micro-architectural evaluation of areas 1 and 2, the porous lesions were much more extensive than revealed by gross examination. Our analysis also revealed that micro-architectural differences were particularly pronounced in younger non-adults. Taken together, the macroscopic examination of CO appears to reflect only the tip of the iceberg, as the micro-architectural changes are much larger than macroscopically identified.

Quinone outside inhibitor (QoI) fungicide resistance in A. alternata populations was reported in Brazil for the first time in 2019, in São Paulo orchards, and the mutation G143A in cytochrome b ( cytb) was found in resistant isolates. Our study investigated the infectious process, production of Reactive Oxygen Species (ROS) and fungal cell death in resistant (QoI-R) and sensitive (QoI-S) Alternaria alternata pathotype tangerine (Aapt) isolates. Morphological characterization of Aapt isolates was performed using Confocal Laser Scanning Microscopy (CLSM). Alternaria brown spot (ABS) symptoms were produced by Aapt isolates on tangelo cv. BRS Piemonte. Germination of QoI-R conidia and production of germ tubes on tangelo leaflets treated with 100µg mL -1 of pyraclostrobin 18 hours after inoculation (hai) was observed using Scanning Electron Microscopy (SEM). At the same time, QoI-S conidial germination was inhibited on tangelo leaflets treated with pyraclostrobin. ROS production and cell death in Aapt isolates at high fungicide concentration were observed using CLSM. QoI-S conidia exhibited high ROS production, indicating high oxidative stress. When dyed with Propidium Iodate (PI), QoI-S conidia emitted red fluorescence, showing cell death and confirming their sensitive phenotype. In contrast, QoI-R conidia neither produced ROS nor exhibited red fluorescence, indicating no cell death and confirming their resistant phenotype. Therefore, our findings evidence that microscopic techniques may help characterize events during fungi-plant interactions, ROS production, cell death, and Aapt phenotypes resistant and sensitive to QoIs using fluorometric protocols. Research Highlights Our findings demonstrate the infection process, reactive oxygen species production, and cell death of Aapt isolates resistant and sensitive to quinone outside inhibitors (QoIs). Confocal laser scanning microscopy may help to characterize phenotypes resistant and sensitive to QoIs using fluorometric protocols.

The Role of the Ocular Lids of the Black-Winged Kite, Elanus caeruleus, in the Immune Protection of the EyeNahed Ahmed Shawki1, Abdelraheim H Attaai2, Amany Mohamed Abdel- Mageed1, Fatma Abdel- Regal Mahmoud11 Assiut University, Faculty of Science. Zoology Department, Assiut, Egypt.2 Department of Anatomy and Embryology, Faculty of Veterinary Medicine, Assiut University, Assiut, Assiut 71526, Egypt.2 Department of Anatomy and Histology, School of Veterinary Medicine, Badr University in Assiut, Assiut 11829, Egypt; abdelraheim.attaai@aun.edu.eg

We present the development of a custom-built structured illumination microscope (SIM) featuring a specially designed transmission diffraction grating. Employing the analogue microfilming method, we fabricated transmission diffraction gratings suited to the specific requirements of our system. This robust and cost-effective method allows for the fabrication of diffraction gratings with customised constants, ensuring excellent transmission in both the visible and near-infrared spectra. Additionally, to assess the performance of our system, we measured the resolution in both epifluorescent and superresolution imaging modalities by applying two independent methods: the conventional resolution measurement using fluorescent beads and the knife-edge technique applied on the MoS 2 monolayer flakes. Both methods confirmed enhancement in the resolution of SIM over the epifluorescent imaging modality. Furthermore, we have successfully demonstrated the capabilities of our microscope by imaging fluorescently labelled astrocytes, specifically targeting the vimentin filament protein within these cells. The superresolution images reveal fine structures of the vimentin cytoskeleton that remain unresolved in the epifluorescent image.

Waveguide evanescent field fluorescence microscopy (WEFF) is an evanescent based microscopy utilizes a confined thin film of light, around 100 nm, to image the plasma membrane of cells attached to a waveguide. Low photobleaching and low background beside its high axial resolution allows time-lapse imaging to investigate changes in cell morphology in the presence or absence of chemical agents. Both large field of view (FOV) and uniform illumination are very important while imaging cell-substrate contacts with an evanescent field. In the current work, we demonstrate that the WEFF microscope is capable of large FOVs with a uniform illumination source and imaging over a very long time period with a simple and an inexpensive experimental setup. The interaction of the trypsin with plasma membranes of live osteoblast cells is investigated. To analyze cell images (250 images), instead of relying on manual tracking, which is time-consuming and can introduce numerous errors, we performed image processing using TrackMate to investigate the dynamic response of cells upon exposure to trypsin. This helps to save time and increase the accuracy of the analysis. The powerful tracking and analysis capabilities of the TrackMate plugin in ImageJ is used to automatically detect the cells boarder and traces each cluster of cells. The reduction in cell area is accompanied by a notable increase in mean intensity, reflecting changes in the intracellular environment. However, the background did not change during the experiment, which proves that the fluorescence material remains attached to the cell membrane and does not leak into the cell medium.

In this study, the kidneys of ground squirrel (hibernated and non-hibernated), rabbit and rat were examined macro and micro anatomically. Kidney morphology was investigated by stereo microscopy, light microscopy and scanning electron microscopy. Triple and immunohistochemical staining were performed for light microscopic examinations, cold crushing and takilon injection into the renal arteries were performed for scanning electron microscopic examinations. The course and branching of intrarenal capillaries, morphologic features from glomeruli to foramina papillaria were demonstrated. The location, position, shape, weight and size of the kidneys of the three species studied were determined. Relative medullary thickness (RMT) and kidney index (Ki) values were calculated from the measurements. Based on the RMT values of the three species, it was concluded that they are mammals belonging to the semi-arid and humid habitat category. The number of nephrons with long-segments were high in the kidneys of all three species. Structural findings suggested that the rat may produce more concentrated urine than the ground squirrel and the ground squirrel may produce more concentrated urine than the rabbit. The mean diameter of the renal corpuscles was 112.5 µm in the hiberne ground squirrel, 137.6 µm in the non-hiberne ground squirrel, 138.1 µm in the rabbit and 137.8 µm in the rat. In hibernated ground squirrels, narrowing of the cavum glomeruli and decrease in renal corpuscle diameters were found. In contrast to ground squirrel and rat kidneys, rabbit kidneys showed the presence of species-specific subcapsular glomeruli. Immunohistochemistry (Anti-Nestin antibody) staining and scanning electron microscope (SEM) images revealed the structure of podocytes in detail. Species-specific area cribrosa configurations were detected in the renal papillae of the kidneys we examined. With this study, new renal morphological findings were obtained in ground squirrel, rabbit and rat.

not-yet-known not-yet-known not-yet-known unknown Aim/Purpose: This study aims to compare the efficacy of locally applied Titanium-Platelet Rich Fibrin (T-PRF) and Leukocyte-Platelet Rich Fibrin (L-PRF) in bone defect healing using micro-CT analysis and histopathological examination in rabbit models. Materials and Methods: Eight healthy male New Zealand rabbits, aged 4-6 months, were utilized. Circular bicortical defects (6 mm diameter) were created at three areas in each rabbit. One of the defects was chosen as the control group received saline solution, while T-PRF and L-PRF were applied to the defect sites in separate experimental groups. Bone regeneration was analyzed using high-resolution micro-CT and histological evaluation post-sacrifice. Statistical analysis assessed the significance of observed differences. Results: Micro-CT analysis revealed significant differences among the groups in bone volume, trabecular thickness, trabecular number, connectivity, and connectivity density (p<0.05). Both T-PRF and L-PRF groups exhibited superior bone parameters compared to controls, with L-PRF showing higher values. Trabecular separation and bone surface area to bone volume ratio did not differ significantly among the groups (p>0.05). Histological examination indicated advanced healing stages in the L-PRF group, suggesting both T-PRF and L-PRF contribute to bone regeneration, with L-PRF demonstrating better effects. Conclusion: The study concludes that while both T-PRF and L-PRF aid in bone regeneration, L-PRF shows superior efficacy. L-PRF is recommended for bone defect healing due to its better performance in enhancing bone parameters.

At the end of 2019, in the city of Wuhan, China, a new highly infectious coronavirus affecting the respiratory tract was announced. It was named SARS-Cov2, causing deaths in the population. In the face of this global health emergency, many medical and protective supplies were quickly depleted, and some others of dubious quality emerged, including face masks for medical and population use. Given this uncertainty, this working group developed a complementary methodology to the wind tunnel, a technique usually used to measure the efficiency of particle passage in certified masks. A gravity-feed airbrush gun operating at a pressure of 60 PSI was used for this purpose. Saline water at a concentration of 20 % NaCl was used as the impact liquid on the face mask, and the deposition time was 1 s, emulating the time of sneezing or coughing. The particles that passed through the face masks were deposited in sample holders for scanning electron microscopy (SEM), where a 0.8 X 0.8 cm slide was placed on their surface and the particles covered with an Au film deposited by sputtering. The sizes of the NaCl particles that were allowed to pass through the face masks were evaluated using field-emission SEM (FSEM). It was found that at 0.5 cm between the surface of the face mask and the deposit surface, distance between mask and airbrush nozzle 2.5 cm and 1 s deposits, the particles allowed to pass through the K95 and KN 95 masks were 85% and 88% respectively in the range from 0.2 – 1-0 µm. Based on this methodology, some face masks used daily by the population were evaluated, and the results are shown in this work .

Scanning electron microscopy (SEM) has significantly advanced morphological studies, particularly in the investigation of fish scale structures. This technique has unveiled intricate architectural details that are crucial for fish identification and classification. In this study, macro- and microscopic analyses were employed to examine the scale morphology of Schizothorax plagiostomus, a vulnerable cyprinid fish from Kashmir, focusing on two body regions (key scales, which were taken from the region below dorsal fin and above lateral line; lateral line scales which were taken from the lateral line). The general scale type observed in Schizothorax plagiostomus was cycloid. Two types of shapes viz., polygonal and cordate were reported in this species. The rostral margin of the scales displayed round and waved forms. The scales exhibited a small and round focus which was antero-centrally positioned. The scales featured narrow or wide grooves (radii) categorized into three types: primary, secondary, and tertiary, present across all four scale fields (anterior, posterior, nuand lateral), forming a tetra-sectioned type. Circuli, arranged in circular patterns around focus were present which were densely placed in anterior and lateral field and widely spaced in posterior field. Notably, lepidonts on the circuli and chromatophores on the posterior margin were absent in this species. These scale characteristics and their morphologies offer a valuable tool for the identification, classification, and phylogenetic analysis of various freshwater fish species and genera.

ABSTRACT Ferula ferulaeoides (Steud.) Korov. (FFK) is an ethnomedicine for the treatment of gastrointestinal diseases. However, the lack of systematic quality control standards has hindered its market and clinical application. The purpose of this study is to carry out pharmacognostic identification and quality evaluation of FFK and to speed up the establishment of its quality control standards. The methods include organoleptic evaluation, microscopic observation, proximate analyses, chemical qualitative analysis, thin-layer chromatography (TLC) fingerprinting, and high-performance liquid chromatography (HPLC). The results showed that the leaves of FFK are light green, and the root is fusiform and thick. The resin is milky white in the fresh state and becomes a yellowish-brown irregular mass after air drying. Under the microscope, the secondary structure of the root was developed, and the resin ducts were mainly distributed in the cortex. The powder contained vessels, oil cells, fibers, starch granules, parenchyma cells, resin ducts, and stone cells, while the resin powder only contained resin mass. The contents of ash, water, extract, and heavy metals in FFK all met the general standards of similar medicinal materials in the Chinese Pharmacopoeia (ChP). Chemical qualitative analysis showed that FFK contained terpenoids, flavonoids, coumarins, and volatile oils. The TLC fingerprint of FFK was established. The results of HPLC showed that the content of ferulic acid in FFK was 0.5518 mg·g -1. This study preliminarily established the pharmacognosy identification standard of FFK and, combined with the quality evaluation of FFK, could lay a foundation for the comprehensive development and utilization of FFK.

math_commands Senna martiana (Benth.) H.S. Irwin & Barneby is endemic to Brazil’s dryland Caatinga domain. The species is known as ”canafistula” in folk medicine and its leaves are used as a laxative. Anatomical and histochemical studies of the species’ vegetative organs (roots, stems, and leaves) were conducted to identify features that would support its taxonomic classification as well as provide quality control for its ethnomedicinal use. Dried fragments from both leaflet surfaces were examined using scanning electron microscopy, and paradermal and transverse sections were used in histochemical tests to confirm the presence of lipids, starch, alkaloids, and nonstructural phenolic compounds. Senna martiana exhibits a striate cuticle on both leaflet surfaces, with straight to curved anticlinal cell walls on the adaxial face and sinuous walls on the abaxial face. The leaflets are amphistomatic, with a variety of stomatal types (paracytic, anisocytic, anomotetracytic, and brachyparacytic). The midrib is plane-convex with a collateral vascular system; the petiole is elliptic with 6–8 vascular bundles; the leaf rachis is ovate with 4–5 vascular bundles. Stems have a siphonostelic ectophloic vascular system. Idioblasts containing crystal sand, prismatic crystals, druses, and starch grains were observed in all vegetative organs. The distinctive combination of leaflet edge contours, leaf rachis and stem structure, and the arrangement and number of vascular bundles provides a set of distinguishing features for Senna martiana that are useful for both taxonomy and quality control of its ethnomedicinal use.

Herein, we adapted a quick, cost-effective, and an environmentally friendly biological method for the synthesis of Ag-NPs using silver nitrate solution as a precursor, and the aqueous leaf extract of Lespedeza juncea (Chinese lespedeza) was exploited to synthesize Ag-NPs. Various physiochemical characterization techniques were used to characterize the nanoparticles. The UV-spectrum revealed a resonance absorption peak at 428 nm, indicating successful synthesis of the nanoparticles. The EDX results indicated the presence of Ag, C, O, and Cl elements in biosynthesized Ag-NPs with elemental composition of 53.93%, 32.34%, 12.49%, and 1.23% respectively. The XRD analysis displayed the crystalline nature of the nanoparticles with face-centered cubic lattice. The FTIR spectrum confirmed the involvement of plant-based biological compounds as reducing and capping agents. TEM revealed quasi-spherical Ag-NPs of 50 nm or smaller in size. An insight of its biological activities reveals significant antimicrobial activities against S. aureus (14.1±0.76 mm), P. aeruginosa (11.50± 0.40 mm), E. coli (16± 0.95 mm), and the fungal strains Viz Candida albicans (14±0.80 mm), Aspergillus flavus (16.50±0.50 mm), Aspergellus niger (17± 0.86 mm) and afflicted 75% mortality to C. eligans. Furthermore, the Ag-NPs were found to be potent inhibitors of the enzymes; tyrosinase, urease, acetylcholinesterase, and butyrylcholinesterase with IC 50 values of 14.3±0.2 μg/mL, 19.5±1.1 μg/mL, 9.3±1.3 μg/mL and 32.65±1.9 μg/mL respectively. The overall outcome of the study suggests that L. jumcea mediated synthesized Ag-NPs hold the potential to be employed as a promising tool for their antibacterial, antifungal, nematocidal and for variety of enzymes inhibitory activities.

Pollen from 20 populations representing 13 Iranian species of the genus Crocus was analyzed using light microscopy (LM) and scanning electron microscopy (SEM) to investigate the palyno-morphological features and their taxonomic significance. Polliniferous materials were sourced from either fresh plants or herbarium samples. For LM analysis, pollen grains were acetolyzed, while intact pollen grains were used for SEM micrographs. The study examined various pollen traits, including polar and equatorial diameters, aperture and meso-aperture width, exine thickness, and P/E ratios in LM analyses. Additionally, SEM observations focused on spinule width and length and the density of spinules and perforations on the exine. The investigation revealed that the pollen grains of Crocus are monads, oblate-spheroidal in shape, and measure approximately 61 to 106.48 micrometers along their equatorial axis. The smallest pollen grains were observed in C. reinhardii, while the largest were found in C. archibaldiorum. The pollen grains were intectate, with the exine irregularly perforated and covered with microechinate ornamentation. Two distinct types of pollen apertures were identified in the studied species: polyaperturoidate apertures in the two populations of Crocus haussknechtii from section Crocus and spiraaperturate pollen in the remaining taxa belonging to section Nudiscapus. Our data suggest that pollen characteristics offer valuable synapomorphies for delimiting sections within the genus Crocus, and also provide significant support for understanding and clarifying relationships within taxonomic aggregates and complexes.

Focused Ion Beam – Scanning Electron Microscopy (FIB-SEM) is a microscopy technique which can be used to investigate the quality and structural properties of industrial materials such as polyolefins. An understudied aspect and possible drawback of the technique could be the implantation of the impinging ions under the sample surface and damage to the molecular structure, hindering its use as a sample preparation tool for surface sensitive techniques. We systematically investigated the damaging effects of gallium liquid metal focused ion beam under grazing incident beam angle and various accelerating voltages on polyethylene. Changes in molecular structure and ion implantation depth were analyzed with Time-of-Flight Secondary Ion Mass Spectrometry depth profiling (ToF-SIMS) and optical profilometry. Our results show that the gallium ion beam causes significant damage to the polyolefin structure, which is especially observed as dehydrogenation of the molecular structure of the (sub)surface. These molecular products are concentrated in distinct sub-surface zones, where damage is coupled to the presence of implanted gallium from the FIB etching.

Biofilms are found in diverse environmental settings and are considered to be responsible for various recalcitrant infections. One characteristic feature of biofilms is resistance to antibiotics, which is the leading cause of recurrent infections and treatment failure. Eradicating the biofilms necessitates the need for agents with promising anti-biofilm potentials. In the present study, the secondary metabolites of the fungal endophyte Cephalotheca foveolata (N11) isolated from the woody tissues of the medicinal plant Teucrium stocksianum were investigated for their antibiofilm potential against the test organisms. For evaluating the antibiofilm activities, in-vitro assays including biofilm inhibition and eradication assays were employed. The bioactive metabolites of the N11 strain exhibited the highest biofilm inhibition and eradication potential of 87.62% and 79.22% respectively against S. epidermidis. The results were further validated by light microscopy and confocal laser scanning microscope which revealed considerable distortion of the biofilm architecture by test agents. Besides, the effect of secondary metabolites on biofilms of test strain was also observed using Raman spectroscopy. The Raman spectra of treated biofilms exhibited a significant reduction in the intensities of the peaks indicating the denaturation and conformational changes in biomolecules. Furthermore, the partial purification of antibiofilm metabolites of N11 was carried out using solvent extraction following TLC and silica column with further characterization done using FTIR. These findings highlight the remarkable potential of bioactive secondary metabolites of endophytic fungi associated with Teucrium stocksianum in disrupting the biofilms thus suggesting that these metabolites can be exploited for manufacturing effective agents against biofilm-associated complications.

The green synthesis of nanoparticles using plant extracts has garnered significant attention as a reliable and sustainable method for producing functional nanomaterials. Among these, zinc oxide (ZnO) nanoparticles are extensively studied for their potential biological applications. In this study, ZnO nanoparticles were synthesized through a green synthesis approach using an aqueous extract of Musa acuminata leaves and zinc acetate as precursors. The resulting nanoparticles were characterized using Ultraviolet–Visible spectroscopy (UV–Vis), Fourier Transform Infrared Spectroscopy (FT-IR), X-Ray Diffraction analysis (XRD), Energy-Dispersive X-ray analysis (EDX), and Scanning Electron Microscopy (SEM). The UV–Vis spectra revealed characteristic absorption peaks around 350 nm, attributed to the nanoparticles’ large excitation binding energy at room temperature. FT-IR analyses confirmed the formation of zinc oxide chemical bonds, while XRD results indicated a hexagonal wurtzite crystal structure. SEM analysis showed that the nanoparticles had a nearly cuboid shape, and EDX analysis confirmed their high purity. The minimum inhibitory concentrations (MIC) of the synthesized ZnO nanoparticles were evaluated against Staphylococcus aureus and E. coli cultures. Cotton wound bandages impregnated with ZnO nanoparticles at concentrations near the calculated MIC demonstrated significant antibacterial activity in vitro. These antimicrobial bandages show potential for use in treating and protecting infection-prone wounds, such as diabetic or burn-related injuries.