Reconstruction of Derogenes varicus Miracidium (Digenea: Derogenidae): First Ultrastructural Description of Spines on the Surface of Hemiurata Larvae.

We performed a detailed ultrastructural reconstruction of the "passive" miracidium of Derogenes varicus Muller, 1784 , a species from Hemiurata group. The miracidium is highly miniaturized and simplified in comparison with the "active" miracidia. For the first time we elucidate the nature of the spines on the surface of hemiuroid larva: they are derivatives of the epithelial plates. The anterior end of the larva is equipped with three epithelial plates that bear both spines and cilia. The major part of the miracidial surface is formed by tegument. The nervous and excretory systems of the D. varicus miracidium are extremely reduced. Single undifferentiated cell comprises the germinal material of the miracidium. We discuss the trends of evolution of hemiuroid miracidia that are associated with transition to passive strategy of infection.

Elucidating the formation of the uniform "glass-like" texture in dried-bonito during processing based on microstructure and protein properties.

Dried-bonito (Katsuobushi) exhibits a unique uniform "glass-like" texture after traditional smoke-drying. Herein, we developed a novel processing method for dried-bonito and elucidated the mechanism of transformation of loose muscle into a "glass-like" texture in terms of texture, microstructure, and protein properties. Our findings showed that the unfolding and aggregation of proteins after thermal induction was a key factor in shaping the "glass-like" texture in bonito muscle. During processing, myofibrils aggregated, the originally alternating thick and thin filaments contracted laterally and aligned into a straight line, and protein cross-linking increased. Secondary structural analysis revealed a reduction in unstable ß-turn content from 26.28% to 15.06%. Additionally, an increase in the content of SS bonds was observed, and the conformation changed from g-g-t to a stable g-g-g conformation, enhanced protein conformational stability. Taken together, our findings provide a theoretical basis for understanding the mechanism of formation of the uniform "glass-like" texture in dried-bonito.

Hierarchically design MoS2/CdNi@RGO ultra-thin hybrid sheet for Photocatalytically degradation of Organic Contaminants fingerprinting in environmental matrices.

The aim of this study was to synthesize effective and economical MoS2/CdNi@rGO photocatalysts and investigate their performance in the degradation of organic pollutants in synthetic effluent. The objective was to assess the characterization results of the synthesized photocatalysts using XRD, SEM/EDS, TEM/HR-TEM, Raman spectrum, and BET isotherm analysis tools. These analyses revealed the good adhesion of MoS2 with rGO and provided insights into the structure and properties of the materials. The results showed that the MoS2/CdNi@rGO photocatalysts exhibited remarkable degradation efficiency for organic pollutants such as Rhodamine-B, erichrome black, and malachite green. The outcomes of the study demonstrated that the MoS2/CdNi@rGO catalyst had the greatest rate constant for Rhodamine-B (RhB) decomposition. which would have been approximately 33 times higher than that of pure RGO (0.0121 min-1). The MoS2/CdNi@rGO photocatalysts also showed excellent recyclability and persistence across five recycle assays, indicating their potential for practical applications in wastewater treatment. The photocatalyst was moderately active, stable up to its fifth usage and stability of the photocatalyst before and after the photocatalytic reaction was also been studied using XRD and SEM. Further research in this area could lead to the development of advanced photocatalytic technologies for environmental remediation.

Thickness determination of hydroperoxidized lipid bilayers from medium-resolution cryo-TEM images.

As the primary products of lipid oxidation, lipid hydroperoxides constitute an important class of lipids generated by aerobic metabolism. However, despite several years of effort, the structure of the hydroperoxidized bilayer has not yet been observed under electron microscopy. Here we use a 200 kV Cryo-TEM to image small unilamellar vesicles (SUVs) made (i) of pure POPC or SOPC, (ii) of their pure hydroperoxidized form, and (iii) of their equimolar mixtures. We show that the challenges posed by the determination of the thickness of the hydroperoxidized bilayers under these observation conditions can be addressed by an image analysis method that we developed and describe here.

In planta synthesis of silver nanoparticles and its effect on adventitious shoot growth and withanolide production in Withania somnifera (L.) Dunal.

Silver (Ag) is a non-essential heavy metal with substantial environmental toxicity but an excellent promotor for plant organogenesis. It is used as an elicitor for secondary metabolite production and for in planta synthesis of metal nanoparticles (MNPs). In the present study, the Ag accumulation and reduction capability of in vitro shoots of Withania somnifera and the toxicity and elicitation effect of Ag on in vitro shoots were explored. In vitro shoot cultures of W. somnifera were treated with different concentrations of silver nitrate for a specific treatment period. Growth index, withaferin A, elemental and electron microscopy analyses were done on silver-treated in vitro shoots of W. somnifera. 1 mM silver nitrate treatment for 12 days period was found to give increased growth index (1.425 ± 0.05c) and withaferin A (2.568 ± 0.08e mg g-1) content. The concentration of bioaccumulated Ag in 1 mM silver nitrate treated in vitro shoot was found to be 50.8 ppm. The presence of nano-Ag was also found in the leaves of 1 mM silver nitrate-treated in vitro shoots. In summary, this is the first report portraying the bioaccumulation and in planta reduction capability of the in vitro shoot system of W. somnifera, which makes it a potential medicinal plant of commercial value for silver contaminated soils.

Operando Electrodeposition of Nonprecious Metal Copper Nanocatalysts on Low-Dimensional Support Materials for Nitrate Reduction Reactions.

Supported nonprecious metal catalysts such as copper (Cu) are promising replacements for Pt-based catalysts for a wide range of energy-related electrochemical reactions. Direct electrochemical deposition is one of the most straightforward and versatile methods to synthesize supported nonprecious metal catalysts. However, further advancement in the design of supported nonprecious metal catalysts requires a detailed mechanistic understanding of the interplay between kinetics and thermodynamics of the deposition phenomena under realistic reaction conditions. Here, we study the electrodeposition of Cu on carbon nanotubes and graphene derivatives under electrochemical conditions using in situ liquid cell transmission electron microscopy (TEM). By combining real-time imaging, electrochemical measurements, X-ray photoelectron spectroscopy (XPS), and finite-element analysis (FEA), we show that low-dimensional support materials, especially carbon nanotubes, are excellent for generating uniform and finely dispersed platinum group metal-(PGM)-free catalysts under mild electrochemical conditions. The electrodeposited Cu on graphene and carbon nanotubes is also observed to show good electrochemical activity toward nitrate reduction reactions (NO3RRs), further supported by density functional theory (DFT) calculations. Nitrogen doping plays an important role in guiding nonprecious metal deposition, but its low electrical conductivity may give rise to lower NO3RR activity compared to its nondoped analogue. The development of supported nonprecious metals through interfacial and surface engineering for the design of supported catalysts will substantially reduce the demand for precious metals and generate robust catalysts with better durability, thereby presenting opportunities for solving the critical problems in energy storage and electrocatalysis.

Influence of loss function and electron dose on ptychography of 2D materials using the Wirtinger flow.

Iterative phase retrieval is based on minimising a loss function as a measure of the consistency of an initial guess and underlying experimental data. Under ideal experimental conditions, real data contains Poissonian noise due to counting statistics. In this work, we use the Wirtinger Flow concept in combination with four common loss functions, being the L1 loss, the mean-squared error (MSE), the amplitude loss and the Poisson loss. Since only the latter reflects the counting statistics as an asymmetric Poisson distribution correctly, our simulation study focuses on two main cases. Firstly, high-dose momentum-resolved scanning transmission electron microscopy (STEM) of an MoS2 monolayer is considered for phase retrieval. In this case, it is found that the four losses perform differently with respect to chemical sensitivity and frequency transfer, which we interprete in terms of the substantially different signal level in the bright and dark field part of diffraction patterns. Remedies are discussed using further simulations, addressing the use of virtual ring detectors for the dark field, or restricting loss calculation to the bright field. Secondly, a dose series is presented down to 100 electrons per diffraction pattern. It is found that all losses yield qualitatively reasonable structural data in the phase, whereas only MSE and Poisson loss range at the correct amplitude level. Chemical contrast is, in general, reliably obtained using the Poisson concept, which also provides the most continuous spatial frequency transfer as to the reconstructed object transmission function.

Formation of {111} oriented domains during the sputtering epitaxy growth of (001) oriented Iridium films.

In the wafer-scale growth of Ir(001) on yttria-stabilized zirconia (YSZ) by magnetron sputtering epitaxy two kinds of {111} oriented domains are observed. One consists of sharp 'fjord'-shaped features in which four 90° alternated rotational variants of {111} are possible and the second one consists of islands with less defined shapes in which eight 45° alternated rotational variants can be found. Their formation occurs directly at the Ir/YSZ interface along incoherent grain boundaries, likely nucleating at local defects of the YSZ surface. In order to avoid these misoriented domains, process separation and proper etching pretreatment of the wafers both before and between the sputtering processes have been found to be the key strategy for achieving reproducibility and overall better material quality.

Enzyme-triggered assembly of glycan nanomaterials.

A comprehensive molecular understanding of carbohydrate aggregation is key to optimize carbohydrate utilization and to engineer bioinspired analogues with tailored shape1s and properties. However, the lack of well-defined synthetic standards has substantially hampered advances in this field. Herein, we employ a phosphorylation-assisted strategy to synthesize previously inaccessible long oligomers of cellulose, chitin, and xylan. These oligomers were subjected to enzyme-triggered assembly (ETA) for the on-demand formation of well-defined carbohydrate nanomaterials, including elongated platelets, helical bundles, and hexagonal particles. Cryo-electron microscopy and electron diffraction analysis provided molecular insights into the aggregation behavior of these oligosaccharides, establishing a direct connection between the resulting morphologies and the oligosaccharide primary sequence. Our findings demonstrate that ETA is a powerful approach to elucidate the intrinsic aggregation behavior of carbohydrates in nature. Moreover, the ability to access a diverse array of morphologies, expanded with a non-natural sequence, underscores the potential of ETA, coupled with sequence design, as a robust tool for accessing programmable glycan architectures.

Monitoring mouse papillomavirus-associated cancer development using longitudinal Pap smear screening.

A substantial percentage of the population remains at risk for cervical cancer due to pre-existing human papillomavirus (HPV) infections, despite prophylactic vaccines. Early diagnosis and treatment are crucial for better disease outcomes. The development of new treatments heavily relies on suitable preclinical model systems. Recently, we established a mouse papillomavirus (MmuPV1) model that is relevant to HPV genital pathogenesis. In the current study, we validated the use of Papanicolaou (Pap) smears, a valuable early diagnostic tool for detecting HPV cervical cancer, to monitor disease progression in the MmuPV1 mouse model. Biweekly cervicovaginal swabs were collected from the MmuPV1-infected mice for viral DNA quantitation and cytology assessment. The Pap smear slides were evaluated for signs of epithelial cell abnormalities using the 2014 Bethesda system criteria. Tissues from the infected mice were harvested at various times post-viral infection for additional histological and virological assays. Over time, increased viral replication was consistent with higher levels of viral DNA, and it coincided with an uptick in epithelial cell abnormalities with higher severity scores noted as early as 10 weeks after viral infection. The cytological results also correlated with the histological evaluation of tissues harvested simultaneously. Both immunocompromised and immunocompetent mice with squamous cell carcinoma (SCC) cytology also developed vaginal SCCs. Notably, samples from the MmuPV1-infected mice exhibited similar cellular abnormalities compared to the corresponding human samples at similar disease stages. Hence, Pap smear screening proves to be an effective tool for the longitudinal monitoring of disease progression in the MmuPV1 mouse model. IMPORTANCE: Papanicolaou (Pap) smear has saved millions of women's lives as a valuable early screening tool for detecting human papillomavirus (HPV) cervical precancers and cancer. However, more than 200,000 women in the United States alone remain at risk for cervical cancer due to pre-existing HPV infection-induced precancers, as there are currently no effective treatments for HPV-associated precancers and cancers other than invasive procedures including a loop electrosurgical excision procedure (LEEP) to remove abnormal tissues. In the current study, we validated the use of Pap smears to monitor disease progression in our recently established mouse papillomavirus model. To the best of our knowledge, this is the first study that provides compelling evidence of applying Pap smears from cervicovaginal swabs to monitor disease progression in mice. This HPV-relevant cytology assay will enable us to develop and test novel antiviral and anti-tumor therapies using this model to eliminate HPV-associated diseases and cancers.

Application of Weak-Beam Dark-Field STEM for Dislocation Loop Analysis †.

Nanoscale dislocation loops formed by irradiation can significantly contribute to both irradiation hardening and embrittlement of materials when subjected to extreme nuclear reactor environments. This study explores the application of weak-beam dark-field (WBDF) scanning transmission electron microscopy (STEM) methods for quantitative irradiation-induced defect analysis in crystalline materials, with a specific focus on dislocation loop imaging and analysis. A high-purity Fe-5 wt% Cr model alloy was irradiated with 8 MeV Fe2+ ions at 450°C to a fluence of 8.8 × 1019 m-2, inducing dislocation loops for analysis. While transmission electron microscopy (TEM) has traditionally been the primary tool for dislocation imaging, recent advancements in STEM technology have reignited interest in using STEM for defect imaging. This study introduces and compares three WBDF STEM methods, demonstrating their effectiveness in suppressing background contrasts, isolating defect information for dislocation loop type classification, providing finer dislocation line images for small loop analysis, and presenting inside-outside contrast for identifying loop nature. Experimental findings indicate that WBDF STEM methods surpass traditional TEM approaches, yielding clearer and more detailed images of dislocation loops. The study concludes by discussing the potential applications of WBDF STEM techniques in defect analysis, emphasizing their adaptability across various material systems beyond nuclear materials.

Structural Complexities in Sodium Ion Conductive Antiperovskite Revealed by Cryogenic Transmission Electron Microscopy.

We use low-dose cryogenic transmission electron microscopy (cryo-TEM) to investigate the atomic-scale structure of antiperovskite Na2NH2BH4 crystals by preserving the room-temperature cubic phase and carefully monitoring the electron dose. Via quantitative analysis of electron beam damage using selected area electron diffraction, we find cryogenic imaging provides 6-fold improvement in beam stability for this solid electrolyte. Cryo-TEM images obtained from flat crystals revealed the presence of a new, long-range-ordered supercell with a cubic phase. The supercell exhibits doubled unit cell dimensions of 9.4 Å × 9.4 Å as compared to the cubic lattice structure revealed by X-ray crystallography of 4.7 Å × 4.7 Å. The comparison between the experimental image and simulated potential map indicates the origin of the supercell is a vacancy ordering of sodium atoms. This work demonstrates the potential of using cryo-TEM imaging to study the atomic-scale structure of air- and electron-beam-sensitive antiperovskite-type solid electrolytes.

Ultrastructural Changes of Synapses in the Hippocampus of Sprague Dawley Rat Brain following Exposure to Naphthalene Balls.

Introduction: The synaptic contacts play an important role in central nervous system (CNS) functioning. Ultrastructural features of synapses in CNS are not studied in naphthalene neurotoxicity model. Materials and Methodology: In the present work, transmission electron microscopy was used for studying the ultrastructural features of synapses in the hippocampus of Sprague Dawley rat brain, on subsequent exposure to naphthalene balls. The ultrastructural changes were observed for naphthalene low dose (200 mg), high dose (400 mg) after the treatment for 28 days, and post-delayed toxicity phase after 14 days in Sprague Dawley rats. Results: In comparison with different groups of naphthalene exposure including control and satellite, axon degeneration, axonal demyelination and abnormal synapses was observed in high dose naphthalene administration group. In the post-delayed naphthalene toxicity group, degeneration of synaptic contacts was observed. Conclusions: This exploration of ultrastructural variations in the synapses of Hippocampus gives information that will be valued in naphthalene neurotoxicological research.

Development and Fabrication of a Multi-Layer Planar Solar Light Absorber Achieving High Absorptivity and Ultra-Wideband Response from Visible Light to Infrared.

The objective of this study is to create a planar solar light absorber that exhibits exceptional absorption characteristics spanning from visible light to infrared across an ultra-wide spectral range. The eight layered structures of the absorber, from top to bottom, consisted of Al2O3, Ti, Al2O3, Ti, Al2O3, Ni, Al2O3, and Al. The COMSOL Multiphysics® simulation software (version 6.0) was utilized to construct the absorber model and perform simulation analyses. The first significant finding of this study is that as compared to absorbers featuring seven-layered structures (excluding the top Al2O3 layer) or using TiO2 or SiO2 layers as substituted for Al2O3 layer, the presence of the top Al2O3 layer demonstrated superior anti-reflection properties. Another noteworthy finding was that the top Al2O3 layer provided better impedance matching compared to scenarios where it was absent or replaced with TiO2 or SiO2 layers, enhancing the absorber's overall efficiency. Consequently, across the ultra-wideband spectrum spanning 350 to 1970 nm, the average absorptivity reached an impressive 96.76%. One significant novelty of this study was the utilization of various top-layer materials to assess the absorption and reflection spectra, along with the optical-impedance-matching properties of the designed absorber. Another notable contribution was the successful implementation of evaporation techniques for depositing and manufacturing this optimized absorber. A further innovation involved the use of transmission electron microscopy to observe the thickness of each deposition layer. Subsequently, the simulated and calculated absorption spectra of solar energy across the AM1.5 spectrum for both the designed and fabricated absorbers were compared, demonstrating a match between the measured and simulated results.

High-Throughput Micro-Combinatorial TEM Phase Mapping of the DC Magnetron Sputtered YxTi1-xOy Thin Layer System.

High-throughput methods are extremely important in today's materials science, especially in the case of thin film characterization. The micro-combinatorial method enables the deposition and characterization of entire multicomponent thin film systems within a single sample. In this paper, we report the application of this method for the comprehensive TEM characterization of the Y-Ti-O layer system. Variable composition samples (YxTi1-xOy) were prepared by dual DC magnetron sputtering, covering the entire (0 ≤ x ≤ 1) concentration range. The structure and morphology of phases formed in both as-deposited and annealed samples at 600, 700, and 800 °C were revealed as a function of Y-Ti composition (x). A comprehensive map showing the appropriate amorphous and crystalline phases, and their occurrence regions of the whole Y-Ti-O layer system, was revealed. Thanks to the applied method, it was shown with ease that at the given experimental conditions, the Y2Ti2O7 phase with a pyrochlore structure forms already at 700 °C without the TiO2 and Y2O3 by-phases, which is remarkably lower than the required temperature for most physical preparation methods, demonstrating the importance and benefits of creating phase maps in materials science and technology.

Nanoscale gold nanoparticle (GNP)-laden tumor cell model and its use for estimation of intracellular dose from GNP-induced secondary electrons.

BACKGROUND: Gold nanoparticles (GNPs) accumulated within tumor cells have been shown to sensitize tumors to radiotherapy. From a physics point of view, the observed GNP-mediated radiosensitization is due to various downstream effects of the secondary electron (SE) production from internalized GNPs such as GNP-mediated dose enhancement. Over the years, numerous computational investigations on GNP-mediated dose enhancement/radiosensitization have been conducted. However, such investigations have relied mostly on simple cellular geometry models and/or artificial GNP distributions. Thus, it is at least desirable, if not necessary, to conduct further investigations using cellular geometry models that properly reflect realistic cell morphology as well as internalized GNP distributions at the nanoscale. PURPOSE: The primary aim of this study was to develop a nanometer-resolution geometry model of a GNP-laden tumor cell for computational investigations of GNP-mediated dose enhancement/radiosensitization. The secondary aim was to demonstrate the utility of this model by quantifying GNP-induced SE tracks/dose distribution at sub-cellular levels for further validation of a nanoscopic dose point kernel (nDPK) method against full-fledged Geant4 Monte Carlo (MC) simulation. METHODS: A transmission electron microscopy (TEM) image of a single cell showing cytoplasm, cellular nucleus, and internalized GNPs in the cellular endosome was segmented into sub-cellular levels based on pixel value thresholding. A corresponding material density was allocated to each pixel, and, by adding a thickness, each pixel was transformed to a geometric voxel and imported as a Geant4-acceptable input geometry file. In Geant4-Penelope MC simulation, a clinical 6 MV photon beam was applied, vertically or horizontally to the cell surface, and energy deposition to the cellular nucleus and cytoplasm, due to SEs emitted by internalized GNPs, was scored. Next, nDPK calculations were performed by generating virtual electron tracks from each GNP voxel to all nucleus and cytoplasm voxels. Subsequently, another set of Geant4 simulation was performed with both Penelope and DNA physics models under the geometry closely mimicking in vitro cell irradiation with a clinical 6 MV photon beam, allowing for derivation of nDPK specific to this geometry and further comparison between Gean4 simulation and nDPK method. RESULTS: The Geant4-calculated SE tracks and associated energy depositions showed significant dependence on photon incidence angle. For perpendicular incidence, nDPK results showed good agreement (average percentage pixel-to-pixel difference of 0.4% for cytoplasm and 0.5% for nucleus) with Geant4 results, while, for parallel incidence, the agreement became worse (-1.7%-0.7% for cytoplasm and -5.5%-0.8% for nucleus). Under the 6 MV cell irradiation geometry, nDPK results showed reasonable agreement (pixel-to-pixel Pearson's product moment correlation coefficient of 0.91 for cytoplasm and 0.98 for nucleus) with Geant4 results. CONCLUSIONS: The currently developed TEM-based model of a GNP-laden cell offers unprecedented details of realistic intracellular GNP distributions for nanoscopic computational investigations of GNP-mediated dose enhancement/radiosensitization. A benchmarking study performed with this model showed reasonable agreement between Geant4- and nDPK-calculated intracellular dose deposition by SEs emitted from internalized GNPs, especially under perpendicular incidence - a popular cell irradiation geometry and when the Geant4-Penelope physics model was used.

Mass mortality of Cornu aspersum in Italian snail farms: A histopathological survey.

The production demand of edible snails in the Mediterranean area is very high and the attention to snail borne diseases is increasing. Following mass mortality events, we have analyzed 240 samples of Cornu aspersum collected from farms across Italy. Anatomopathological examination showed the presence of alterations of the gastro-intestinal apparatus and of the digestive gland, while histopathological examination revealed the presence of Rickettsia-like organisms (RLOs) in 70% (168/240) of cases and Giemsa positive amoebae in the remaining 30% (72/240) of cases. RLOs were localized mainly at the level of the DG, where regressive changes or nodular inflammation was observed. TEM examination of RLOs samples revealed the presence of many rod-shaped electron dense microorganisms. Amoebal infection occurred in the kidney, intestine, lung, the DG and were associated to regressive events or infiltrative/nodular and encapsulation like inflammation. To date it is still unclear if the pathogens detected could represent a risk for humans and animals, therefore further studies are needed to better elucidate this point.

Characterization of the clinically approved MRI tracer resotran for magnetic particle imaging in a comparison study.

Objective.The availability of magnetic nanoparticles (MNPs) with medical approval for human intervention is fundamental to the clinical translation of magnetic particle imaging (MPI). In this work, we thoroughly evaluate and compare the magnetic properties of an magnetic resonance imaging (MRI) approved tracer to validate its performance for MPI in future human trials.Approach.We analyze whether the recently approved MRI tracer Resotran is suitable for MPI. In addition, we compare Resotran with the previously approved and extensively studied tracer Resovist, with Ferrotran, which is currently in a clinical phase III study, and with the tailored MPI tracer Perimag.Main results.Initial magnetic particle spectroscopy (MPS) measurements indicate that Resotran exhibits performance characteristics akin to Resovist, but below Perimag. We provide data on four different tracers using dynamic light scattering, transmission electron microscopy, vibrating sample magnetometry measurements, MPS to derive hysteresis, point spread functions, and a serial dilution, as well as system matrix based MPI measurements on a preclinical scanner (Bruker 25/20 FF), including reconstructed images.Significance.Numerous approved MNPs used as tracers in MRI lack the necessary magnetic properties essential for robust signal generation in MPI. The process of obtaining medical approval for dedicated MPI tracers optimized for signal performance is an arduous and costly endeavor, often only justifiable for companies with a well-defined clinical business case. Resotran is an approved tracer that has become available in Europe for MRI. In this work, we study the eligibility of Resotran for MPI in an effort to pave the way for human MPI trials.

The effects of the combination of temozolomide and Eribulin on T98G human glioblastoma cell line: an ultrastructural study.

Glioblastoma tumors are the most aggressive primary brain tumors that develop resistance to temozolomide (TMZ). Eribulin (ERB) exhibits a unique mechanism of action by inhibiting microtubule dynamics during the G2/M cell cycle phase. We utilized the T98G human glioma cell line to investigate the effects of ERB and TMZ, both individually and in combination. The experimental groups were established as follows: control, E5 (5 nM ERB), T0.75 (0.75 mM TMZ), T1 (1.0 mM TMZ), and combination groups (E5+T0.75 and E5+T1). All groups showed a significant decrease in cell proliferation. Apoptotic markers revealed a time-dependent increase in annexin-V expression, across all treatment groups at the 48-hour time point. Caspase-3, exhibited an increase in the combination treatment groups at the 48-hour mark. Transmission electron microscopy (TEM) revealed normal ultrastructural features in the glioma cells of the control group. However, treatments induced ultrastructural changes within the spheroid glioblastoma model, particularly in the combination groups. These changes included a dose-dependent increase in autophagic vacuoles and apoptotic morphology of the cells. In conclusion, the similarity in the mechanism of action between ERB and TMZ suggests the potential for synergistic effects when combined. Our results highlight that this combination induced severe damage and autophagy in glioma spheroids after 48 hours.

Ultrastructural Organization and Metal Elemental Composition of the Mandibles in Two Ladybird Species.

The mandibles are among the most important appendages of insects' mouthparts. Their morpho-functional organization is correlated with the variation in dietary preferences. In this study, we investigated the ultrastructural organization and metal composition of the mandibles of two ladybird species with different dietary habits: Harmonia axyridis (an entomophagous species) and Subcoccinella vigintiquatuorpunctata (a phytophagous species). The ultrastructural organization was studied using Scanning and Transmission Electron Microscopy, whereas the metal composition was investigated using Energy-Dispersive X-ray spectroscopy (EDX). Significant differences were observed in the general organization and metal enrichment pattern between the two species. The mandibles of H. axyridis are large and present a molar part with two teeth, with the apical one showing a bifid apex. In contrast, S. vigintiquatuorpunctata exhibited a molar region with several teeth on its apical part. The study revealed significant differences in metal content between the teeth and the prostheca of H. axyridis. Mn was the most abundant element in teeth, whereas Cl was more abundant in the prostheca. In the case of S. vigintiquatuorpunctata, Si was the most abundant element in the prostheca, while Mn was more present in the teeth. A comparison between the two species revealed that both teeth and prostheca showed significant variation in the elemental composition. These findings underscore the role of dietary preferences in shaping the structural and metal composition variations in the mandibles of these two ladybird species.