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Antibody (Ab)-based imaging techniques rely on reagents whose performance may be application specific. Because commercial antibodies are validated for only a few purposes, users interested in other applications may have to perform extensive in-house antibody testing. Here, we present a novel application-specific proxy screening step to efficiently identify candidate antibodies for array tomography (AT), a serial section volume microscopy technique for high-dimensional quantitative analysis of the cellular proteome. To identify antibodies suitable for AT-based analysis of synapses in mammalian brain, we introduce a heterologous cell-based assay that simulates characteristic features of AT, such as chemical fixation and resin embedding that are likely to influence antibody binding. The assay was included into an initial screening strategy to generate monoclonal antibodies that can be used for AT. This approach simplifies the screening of candidate antibodies and has high predictive value for identifying antibodies suitable for AT analyses. In addition, we have created a comprehensive database of AT-validated antibodies with a neuroscience focus and show that these antibodies have a high likelihood of success for postembedding applications in general, including immunogold electron microscopy. The generation of a large and growing toolbox of AT-compatible antibodies will further enhance the value of this imaging technique.

Lindenia tetraphylla (Van der Linden, 1825) eggs exhibit an egg structure that is very rare in other Gomphidae species. They have a well‐developed surface reticulation structure. The anterior pole of the egg has a small, rounded micropylar area consisting of seven orifices arranged radially around a central area. The posterior pole has a sessile, truncated cone that carries 55–65 coiled filaments. The filament structure found at the posterior pole of the egg has been observed in the gomphid species Lestinogomphus africanus (Fraser, 1926), Ictinogomphus australis (Selys, 1873), and I. rapax (Rambur, 1842). However, L. tetraphylla eggs differ from these species in both morphology and filament structure. This study provides a detailed analysis of the ultrastructure of L. tetraphylla eggs using scanning electron microscopy, and the functional and taxonomic significance of the eggshell are discussed. Research Highlights: The aim of this study is to examine the ultrastructure of the L. tetraphylla eggshell, emphasizing its function and taxonomic value. In this context, the general morphology of the egg, the reticulations on its surface, the micropylar region and micropylar structure, and the posterior filament coil were examined.In this study, the ultrastructure of L. tetraphylla egg was examined for the first time using scanning electron microscopy (SEM). As a result of the examination, it was detected for the first time that the posterior filament coil, which is rarely seen in odonate eggs, is also present in L. tetraphylla eggs.By comparing the L. tetraphylla egg with the eggs of I. ferox, I. rapax, I. australis, and L. africanus species, which are similar in terms of the posterior filament coil, the features that distinguish the L. tetraphylla egg. [ABSTRACT FROM AUTHOR]

Simple Summary: A decrease in eggshell quality in the late laying period leads to eggshell breakage, seriously affecting production efficiency. Guinea fowl with relatively high eggshell strength have the same layers of eggshell structure and various identical proteins as laying hens. However, their eggshell structures have not been systematically compared. Therefore, this study systematically compared the eggshell ultrastructure, bubble pore index, and composition of laying hens and Guinea fowl. The differences in the eggshell ultrastructure, bubble pores, and composition revealed, to some extent, the structure and composition necessary to increase the eggshell quality of laying hens. This study provides a reference for improving eggshell quality. The decrease in eggshell quality seriously affects production efficiency. Guinea fowl (GF) eggs possess strong eggshells because of their unique crystal structure, and few systematic studies have compared laying hen and GF eggs. Sixty eggs were collected from both 40-week-old Dwarf Layer-White (DWL-White) laying hens and GF, and the eggshell quality, ultrastructure, bubble pores, and composition were measured. The results showed that the DWL-White eggs had a higher egg weight and a lower eggshell strength, strength per unit weight, thickness, and ratio than the GF eggs (p < 0.01). There were differences in the mammillary layer thickness ratio, the effective layer thickness ratio, the quantity of bubble pores (QBPs), the ratio of the sum of the area of bubble pores to the area of the eggshell in each image (ARBE), and the average area of bubble pores (AABPs) between the DWL-White and GF eggs (p < 0.01). The composition analysis demonstrated that there were differences in the organic matter, inorganic matter, calcium, and phosphorus between the DWL-White and GF eggs (p < 0.01). There were positive associations between the mammillary knob number in the image and the QBPs and ARBE and a negative correlation with the AABPs in the DWL-White eggs (p < 0.01). This study observed distinctions that offer new insights into enhancing eggshell quality. [ABSTRACT FROM AUTHOR]

ROS-dependent induction of oxidative damage can be used as a trigger initiating genetically determined non-specific protection in plant cells and tissues. Plants are potentially able to withstand various specific (toxic, osmotic) factors of abiotic effects, but do not have sufficient or specific sensitivity to form an adequate effective response. In this work, we demonstrate one of the possible approaches for successful cold acclimation through the formation of effective protection of photosynthetic structures due to the insertion of the heterologous FeSOD gene into the tobacco genome under the control of the constitutive promoter and equipped with a signal sequence targeting the protein to plastid. The increased enzymatic activity of superoxide dismutase in the plastid compartment of transgenic tobacco plants enables them to tolerate the oxidative factor of environmental stresses scavenging ROS. On the other hand, the cost of such resistance is quite high and, when grown under normal conditions, disturbs the arrangement of the intrachloroplastic subdomains leading to the modification of stromal thylakoids, probably significantly affecting the photosynthesis processes that regulate the efficiency of photosystem II. This is partially compensated for by the fact that, at the same time, under normal conditions, the production of peroxide induces the activation of ROS detoxification enzymes. However, a violation of a number of processes, such as the metabolism of accumulation, and utilization and transportation of sugars and starch, is significantly altered, which leads to a shift in metabolic chains. The expected step for further improvement of the applied technology could be both the use of inducible promoters in the expression cassette, and the addition of other genes encoding for hydrogen peroxide-scavenging enzymes in the genetic construct that are downstream in the metabolic chain. [ABSTRACT FROM AUTHOR]

The plant cell wall is an actively reorganized network during plant growth and triggered immunity in response to biotic stress. While the molecular mechanisms managing perception, recognition, and signal transduction in response to pathogens are well studied in the context of damaging intruders, the current understanding of plant cell wall rebuilding and active defense strategies in response to plant virus infections remains poorly characterized. Pectins can act as major elements of the primary cell wall and are dynamic compounds in response to pathogens. Homogalacturonans (HGs), a main component of pectins, have been postulated as defensive molecules in plant–pathogen interactions and linked to resistance responses. This research focused on examining the regulation of selected pectin metabolism components in susceptible (rbohD-, Col-0-TuMV) and resistance (rbohF-, rbohD/F–TuMV) reactions. Regardless of the interaction type, ultrastructural results indicated dynamic cell wall rebuilding. In the susceptible reaction promoted by RbohF, there was upregulation of AtPME3 (pectin methylesterase) but not AtPME17, confirmed by induction of PME3 protein deposition. Moreover, the highest PME activity along with a decrease in cell wall methylesters compared to resistance interactions in rbohD–TuMV were noticed. Consequently, the susceptible reaction of rbohD and Col-0 to TuMV was characterized by a significant domination of low/non-methylesterificated HGs. In contrast, cell wall changes during the resistance response of rbohF and rbohD/F to TuMV were associated with dynamic induction of AtPMEI2, AtPMEI3, AtGAUT1, and AtGAUT7 genes, confirmed by significant induction of PMEI2, PMEI3, and GAUT1 protein deposition. In both resistance reactions, a dynamic decrease in PME activity was documented, which was most intense in rbohD/F–TuMV. This decrease was accompanied by an increase in cell wall methylesters, indicating that the domination of highly methylesterificated HGs was associated with cell wall rebuilding in rbohF and rbohD/F defense responses to TuMV. These findings suggest that selected PME with PMEI enzymes have a diverse impact on the demethylesterification of HGs and metabolism as a result of rboh–TuMV interactions, and are important factors in regulating cell wall changes depending on the type of interaction, especially in resistance responses. Therefore, PMEI2 and PMEI3 could potentially be important signaling resistance factors in the rboh–TuMV pathosystem. [ABSTRACT FROM AUTHOR]

A novel species, Gymnadenia winkeliana, has been identified in the Bucegi Natural Park ROSCI0013, located in the Southern Carpathians of Central Romania. Two moderately sized populations of Gymnadenia winkeliana, totalling 120–140 individuals, were discovered inhabiting the alpine grasslands of the park, situated 2.000 m above sea level. To describe this newly found population as comprehensively as possible, 44 vegetative and floral organs/organ parts were directly studied and measured from living plants. Special attention was focused on the characteristics that proved to have taxonomic significance, particularly those involving distinctive details in the morphology of the leaves, perianth, labellum and gynostemium. A total of 223 characteristics were analysed encompassing the morphology of every organ of the plant, cytology and breeding system. Furthermore, comprehensive taxonomic treatment and description, accompanied by colour photographs illustrating the holotype, are provided. Voucher specimens were deposited at the Herbarium of the University of Agriculture and Veterinary Medicine, Bucharest (USAMVB Herbarium barcode: 40102, NEA); Gymnadenia winkeliana, a (micro)endemic species, is characterized as a putative allogamous, facultatively apomict that significantly differs from other Gymnadenia R.Br. species found in Romania. Notably, it distinguishes itself through its smaller habitus (reaching heights of up to 8–10 cm), its two-coloured, rounded/hemispherical inflorescence displaying a gradient of pink hues in an acropetal fashion (ranging from whitish-pink at the base to vivid-pink at the topmost flowers), and its limited distribution in high-altitude areas, encompassing approximately 8–10 km2 in the central area of the Bucegi Natural Park. This species has been under observation since 2005, with observed population numbers showing a significant increase over time, from ca. 50–55 (counted at the time of its discovery) to 120–140 individuals (counted in June 2023). Additionally, comprehensive information regarding the habitat, ecology, phenology and IUCN conservation assessments of Gymnadenia winkeliana are provided, including maps illustrating its distribution. [ABSTRACT FROM AUTHOR]

Basal cell carcinoma (BCC) is the most common skin cancer. Skin cancers may present either as a non-invasive tumor or an invasive malignancy. The terminology of carcinoma in situ is used when the tumor is either just limited to epidermis or not present as single cells or nests in the dermis. However, currently the terminology superficial BCC is inappropriately used instead of BCC in situ when the skin cancer is limited to epidermis. In this study we compare the pathologic changes of superficial, nodular, and infiltrative BCCs using electron microscopy to identify the ultrastructural characteristics and validate the previously proposed terminology. Three cases of BCC (superficial BCC, nodular BCC, and infiltrative BCC) diagnosed by dermatopathologists at our institute were selected for review. Paraffin block tissues from these cases were sent for electron microscopy studies which demonstrated disruption of basal lamina in both nodular and infiltrative type of BCC, while it remains intact in BCC superficial type after extensive examination. Therefore, similar to other in situ skin cancers, there is no invasion of the neoplasm in superficial BCC into the dermis. Hence, the older term superficial BCC should be appropriately replaced with the newer terminology BCC in situ. [ABSTRACT FROM AUTHOR]

Synovial sarcoma (SS) and solitary fibrous tumor (SFT) are entities with considerable morphological and immunohistochemical similarities that sometimes show a non-confirmatory profile (TLE1 negative, CD34 and focal or negative STAT6 and lack of specific fusion IHC markers), in which the utility ultrastructure is unknown. A cross-sectional, retrospective, analytical, nonexperimental study was carried out by the Department of Pathology of the National Cancer Institute of Mexico (INCan) e from January 1, 2009 to December 31, 2018. With 17 SFT cases with diffuse or focal CD34 and STAT6 positivity and 18 cases of SS with positive FISH molecular test t(X:18) breakapart were studied by electron microscopy of fresh glutaraldehyde fixed or paraffin-embedded tissue. The ultrastructural findings with a significant difference present in the SS were tandem tight junctions, desmosomes and abundance of dilated rough endoplasmic reticulum (RER) cisternae (p < 0.001, 0.003, and 0.001, respectively); while in the (SFT) the presence of abundant glycogen, basal lamina, long and slender cytoplasmic processes, pinocytic vesicles, hemidesmosomes, and/or dense plaques, collagen skein, and microvilli-like buds (p = 0.028, 0.005, and <0.001 for the last five). We then infer that the five distinctive markers of the SFT are the collagen skeins intermingled with cellular processes in a shape of "squid can," and the pinocytic vesicles as they were not observed in any case of SS. Conversely, tandem junctions were not found in any SFT case. Although the presence of multivesicular buds in the SFT was not significant, it had not been previously described. [ABSTRACT FROM AUTHOR]

Ischemia/reperfusion (I/R) injury of sciatic nerve is a serious condition that results in nerve fiber degeneration, and reperfusion causes oxidative injury. Peripheral blood mononuclear cells (PBMNCs) have neuroregenerative power. This study was carried out to evaluate the potential ameliorative effect of PBMNCs on changes induced by I/R injury of the sciatic nerve. Fifty adult male albino rats were divided into donor and experimental groups that were subdivided into four groups: group I (control group), group II received 50 µL PBNMCs once intravenously via the tail vein, group III rubber tourniquet was placed around their Rt hind limb root for 2 hours to cause ischemia, group IV was subjected to limb ischemia as group III, then they were injected with 50 ul PBMNCs as group II before reperfusion. I/R injury showed disorganization of nerve fascicles with wide spaces in between nerve fibers. The mean area of collagen fibers, iNOS immunoexpression, and number of GFAP-positive Schwann cells of myelinated fibers showed a highly significant increase, while a highly significant reduction in the G-ratio and neurofilament immunoexpression was observed. Myelin splitting, invagination, evagination, and myelin figures were detected. PBMNC-treated group showed a marked improvement that was confirmed by histological, immunohistochemical, and ultrastructural findings. [ABSTRACT FROM AUTHOR]

The ultrastructure of the olfactory system of most fossorial rodents remains largely unexplored. This study sought to investigate the functional structure of the olfactory mucosa and olfactory bulb of two species of fossorial rodents that have distinct behaviour and ecology, the East African root rat (RR) and the naked mole rat (NMR). Transmission electron microscopy and scanning electron microscopy were employed. The basic ultrastructural design of the olfactory system of the two species was largely comparable. In both species, the olfactory mucosa comprised an olfactory epithelium and an underlying lamina propria. The olfactory epithelium revealed olfactory knobs, cilia and microvilli apically and sustentancular cells, olfactory receptor neurons and basal cells in the upper, middle and basal zones, respectively. The lamina propria was constituted by Bowman's glands, olfactory nerve bundles and vasculature supported by loose connective tissue. Within the olfactory bulb, intracellular and extracellular structures including cell organelles, axons and dendrites were elucidated. Notable species differences were observed in the basal zone of the olfactory epithelium and on the luminal surface of the olfactory mucosa. The basal zone of the olfactory epithelium of the RR consisted of a single layer of flattened electron‐dense horizontal basal cells while the NMR had juxtaposed electron‐dense and electron‐lucent heterogenous cells, an occurrence seen as being indicative of quiescent and highly proliferative states of the olfactory epithelia in the two species, respectively. The olfactory epithelial surface of the NMR comprised an elaborate cilia network that intertwined extensively forming loop‐like structures whereas in the RR, the surface was rugged and consisted of finger‐like processes and irregular masses. With gross and histological studies showing significant differences in the olfactory structures of the two species, these findings are a further manifestation that the olfactory system of the RR and the NMR have evolved differently to reflect their varied olfactory functional needs. [ABSTRACT FROM AUTHOR]

Background/Objective: Mesenchymal Stromal Cells (MSCs) have been considered a promising treatment for several diseases, such as cardiac injuries. Many studies have analyzed their functional properties; however, few studies have characterized MSCs through successive culture passages. The main objective of this work was to analyze the phenotype and functionality of MSCs isolated from two different sources in five culture passages to determine if the culture passage might influence the efficacy of MSCs as a cell therapy treatment. Methods: Bone Marrow (BM)-MSCs were harvested from the femur of Wistar rats (n = 17) and Adipose Tissue(AT)-MSCs were isolated from inguinal fat (n = 17). MSCs were cultured for five culture passages, and the immunophenotype was analyzed by flow cytometry, the functionality was characterized by adipogenic, osteogenic, and chondrogenic differentiation assays, and cytokine secretion capacity was determined through the quantification of the Vascular Endothelial Growth-Factor, Fibroblast Growth-Factor2, and Transforming Growth-Factorβ1 in the cell supernatant. The ultrastructure of MSCs was analyzed by transmission electron microscopy. Results: BM-MSCs exhibited typical phenotypes in culture passages two, four, and five, and their differentiation capacity showed an irregular profile throughout the five culture passages analyzed. AT-MSCs showed a normal phenotype and differentiation capacity in all the culture passages. BM- and AT-MSCs did not modify their secretion ability or ultrastructural morphology. Conclusions: Throughout the culture passages, BM-MSCs, but not AT-MSCs, exhibited changes in their functional and phenotypic characteristic that might affect their efficacy as a cell therapy treatment. Therefore, the culture passage selected should be considered for the application of MSCs as a cell therapy treatment. [ABSTRACT FROM AUTHOR]

Telocytes (TCs) are CD34-positive interstitial cells that have long cytoplasmic projections, called telopodes; they have been identified in several organs and in various species. These cells establish a complex communication network between different stromal and epithelial cell types, and there is growing evidence that they play a key role in physiology and pathology. In many tissues, TC network impairment has been implicated in the onset and progression of pathological conditions, which makes the study of TCs of great interest for the development of novel therapies. In this review, we summarise the main methods involved in the characterisation of these cells as well as their inherent difficulties and then discuss the functional assays that are used to uncover the role of TCs in normal and pathological conditions, from the most traditional to the most recent. Furthermore, we provide future perspectives in the study of TCs, especially regarding the establishment of more precise markers, commercial lineages and means for drug delivery and genetic editing that directly target TCs. [ABSTRACT FROM AUTHOR]

The characterization of colleters in Rubiaceae is crucial for understanding their role in plant function. Analyzing colleters in Palicourea tetraphylla and Palicourea rudgeoides aims to deepen the understanding of these structures morphoanatomical and functional characteristics. The study reveals colleters with palisade epidermis and a parenchymatic central axis, classified as standard type, featuring vascularization and crystals. Colleter secretion, abundant in acidic mucopolysaccharides, proteins, and phenolic compounds, protects against desiccation. The ontogenesis, development, and senescence of the colleters are quite rapid and fulfill their role well in biotic and abiotic protection because these structures are present at different stages of development in the same stipule. Pronounced protrusions on the colleters surface, coupled with the accumulation of secretion in the intercellular and subcuticular spaces, suggest that the secretory process occurs through the wall, driven by pressure resulting from the accumulation of secretion. The microorganisms in the colleters' secretion, especially in microbiota-rich environments such as the Atlantic Forest, provide valuable information about plant–microorganism interactions, such as resistance to other pathogens and organisms and ecological balance. This enhanced understanding of colleters contributes to the role of these structures in the plant and enriches knowledge about biological interactions within specific ecosystems and the family taxonomy. [ABSTRACT FROM AUTHOR]

The brain contains thousands of millions of synapses, exhibiting diverse structural, molecular, and functional characteristics. However, synapses can be classified into two primary morphological types: Gray's type I and type II, corresponding to Colonnier's asymmetric (AS) and symmetric (SS) synapses, respectively. AS and SS have a thick and thin postsynaptic density, respectively. In the cerebral cortex, since most AS are excitatory (glutamatergic), and SS are inhibitory (GABAergic), determining the distribution, size, density, and proportion of the two major cortical types of synapses is critical, not only to better understand synaptic organization in terms of connectivity, but also from a functional perspective. However, several technical challenges complicate the study of synapses. Potassium ferrocyanide has been utilized in recent volume electron microscope studies to enhance electron density in cellular membranes. However, identifying synaptic junctions, especially SS, becomes more challenging as the postsynaptic densities become thinner with increasing concentrations of potassium ferrocyanide. Here we describe a protocol employing Focused Ion Beam Milling and Scanning Electron Microscopy for studying brain tissue. The focus is on the unequivocal identification of AS and SS types. To validate SS observed using this protocol as GABAergic, experiments with immunocytochemistry for the vesicular GABA transporter were conducted on fixed mouse brain tissue sections. This material was processed with different concentrations of potassium ferrocyanide, aiming to determine its optimal concentration. We demonstrate that using a low concentration of potassium ferrocyanide (0.1%) improves membrane visualization while allowing unequivocal identification of synapses as AS or SS. [ABSTRACT FROM AUTHOR]

Sperm development and structure in the wood-inhabiting fungal and plant-feeding nematode, Bursaphelenchus luxuriosae, were studied using transmission electron microscopy to evaluate interspecific similarities and differences of spermatozoa in nematodes. In general, spermatogenesis in B. luxuriosae fits the ‘rhabditid’ pattern supported by morphological and phylogenetic analysis of the order Rhabditida. Spermatocyte development includes formation of complexes of fibrous bodies (FBs) with membranous organelles (MOs), the complexes dissociate in the spermatids into separate components, and the immature sperm contain MOs but lack FBs, which transform into a dense matrix of sperm cytoplasm. The female spermatheca contains mature spermatozoa as bipolar cells subdivided into a pseudopod devoid of organelles and a main cell body containing a nucleus without a nuclear envelope, numerous mitochondria, and peripheral MOs as pouches opening to the exterior via pores. Data on B. luxuriosae are used for analysis of variable quantitative and morphological characteristics of spermatozoa in Aphelenchoidea. General size of spermatozoa and their MOs have little value for comparative analysis. The MO knobbles look uniform in immature spermatozoa of each aphelenchoidid species studied and may be considered as a taxonomically specific ultrastructural feature. The presence or absence of FBs in immature spermatozoa demonstrates the diagnostic value of the aphelenchoidid sperm structure at the species level. Analysis also shows close similarity of spermatozoa in Bursaphelenchus spp. and Caenorhabditis elegans, the model species with comprehensive data on sperm biology. This may be used to identify new ways for the control and suppression of harmful nematode species such as B. xylophilus. [ABSTRACT FROM AUTHOR]

Simple Summary: In 2011, the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident resulted in the release of vast quantities of radioactive substances. In this study, we performed an ultrastructural analysis of the testis of the wild large Japanese field mouse (Apodemus speciosus), inhabiting the area near the ex-evacuation zones of the FDNPP and chronically exposed to various levels of low-dose-rate (LDR) radiation, to evaluate the effects of ionizing radiation on the male reproductive system and the possible consequences for future generations. Our results showed a preserved morphology of seminiferous tubules. However, ultrastructural changes, such as wide intercellular spaces, cytoplasmic vacuolization, vacuolated mitochondria, and increased lipid droplet clusters, were found, which could be linked to the mechanisms related to spermatogenesis. Long-term chronic LDR radiation exposure associated with the FDNPP accident had no pronounced adverse effect on spermatogenesis in A. speciosus, even if testis presented some ultrastructural changes. Our findings could be used in future studies to evaluate the effects of radiation on reproductive health in wild animals. Since the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident, great attention has been paid to the impact of chronic low-dose-rate (LDR) radiation exposure on biological systems. The reproductive system is sensitive to radiation, with implications connected to infertility. We investigated the testis ultrastructure of the wild large Japanese field mouse (Apodemus speciosus) from three areas contaminated after the FDNPP accident, with different levels of LDR radiation (0.29 µSv/h, 5.11 µSv/h, and 11.80 µSv/h). Results showed good preservation of the seminiferous tubules, comparable to the unexposed animals (controls), except for some ultrastructural modifications. Increases in the numerical density of lipid droplet clusters in spermatogenic cells were found at high levels of LDR radiation, indicating an antioxidant activity rising due to radiation recovery. In all groups, wide intercellular spaces were found between spermatogenic cells, and cytoplasmic vacuolization increased at intermediate and high levels and vacuolated mitochondria at the high-level. However, these findings were also related to the physiological dynamics of spermatogenesis. In conclusion, the testes of A. speciosus exposed to LDR radiation associated with the FDNPP accident showed a normal spermatogenesis, with some ultrastructural changes. These outcomes may add information on the reproductive potential of mammals chronically exposed to LDR radiation. [ABSTRACT FROM AUTHOR]

This communication deals with seasonal variation of protozoans of edible ornamental fishes of West Bengal, India. During this survey, 1566 edible ornamental fishes were collected from different fish farms, ponds, and nearby markets of several district of West Bengal namely, Nadia, South 24 Parganas, Hooghly, North 24 Parganas, Birbhum, Purba Barddhaman, and Paschim Barddhaman from November 2019 to December 2022. Three ciliate ectoparasites belonging to the genera Trichodina, Trichodinella, and Tripartiella and three myxozoan parasites belonging to the genera Myxobolus, Thelohanellus, and Kudoa have been identified from edible ornamental fish, namely, Anabas testudineus, Bloch, 1792. Our study shows ciliate parasites have a higher rate of infection than myxozoan parasites. The highest prevalence of ectoparasitic infection has been reported during the post-monsoon season, i.e., from November to February, followed by monsoon season while lowest prevalence has been observed during pre-monsoon season, i.e., March to June, implying that environmental variables play a role in the spread of infections. The identification of these ciliates and myxozoans parasites was done by a widely used staining technique, along with an ultrastructural study of the myxozoan parasites using scanning electron microscopy. The current work also reveals the histopathological alterations in the gill and fin of Anabas testudineus, after infection with myxozoan parasites along with an ultrastructural study using scanning electron microscopy, which has not been previously studied. Therefore, this investigation also allows some insight into the previously unrecorded tissue-level alteration of infected host organs due to parasitic infestation. [ABSTRACT FROM AUTHOR]

The epidermis is a stratified epithelium that forms the outer layer of the skin. It is composed primarily of keratinocytes and is constantly renewed by the proliferation of stem cells and their progeny that undergo terminal differentiation as they leave the basal layer and migrate to the skin surface. Basal keratinocytes rest on a basement membrane composed of an extracellular matrix that controls their fate via integrin‐mediated focal adhesions and hemidesmosomes which are critical elements of the epidermal barrier and promote its regenerative capabilities. The distribution of basal cells with optimal activity provides the basement membrane with its characteristic undulating shape; this configuration disappears with age, leading to epidermal weakness. In this study, we present an in‐depth imaging analysis of basal keratinocyte anchorage in samples of human skin from participants across the age spectrum. Our findings reveal that skin aging is associated with the depletion of hemidesmosomes that provide crucial support for stem cell maintenance; their depletion correlates with the loss of the characteristic basement membrane structure. Atomic force microscopy studies of skin and in vitro experiments revealed that the increase in tissue stiffness observed with aging triggers mechanical signals that alter the basement membrane structure and reduce the extent of basal keratinocyte anchorage, forcing them to differentiate. Genomic analysis revealed that epidermal aging was associated with mechanical induction of the transcription factor Krüppel‐like factor 4. The altered mechanical properties of tissue being a new hallmark of aging, our work opens new avenues for the development of skin rejuvenation strategies. [ABSTRACT FROM AUTHOR]

Thick‐walled rosette‐like snow algae were long thought to be a life stage of various other species of snow algae. Rosette‐like cells have not been cultured, but by manually isolating cells from 38 field samples in southern British Columbia, we assigned a variety of rosette morphologies to DNA sequence. Phylogenetic analysis of Rubisco large‐subunit (rbcL) gene, ribosomal internal transcribed spacer 2 (ITS2) rRNA region, and 18S rRNA gene revealed that the rosette‐like cells form a new clade within the phylogroup Chloromonadinia. Based on these data, we designate a new genus, Rosetta, which comprises five novel species: R. castellata, R. floranivea, R. stellaria, R. rubriterra, and R. papavera. In a survey of 762 snow samples from British Columbia, we observed R. floranivea exclusively on snow overlying high‐elevation glaciers, whereas R. castellata was observed at lower elevations, near the tree line. The other three species were rarely observed. Spherical red cells enveloped in a thin translucent sac were conspecific with Rosetta, possibly a developmental stage. These results highlight the unexplored diversity among snow algae and emphasize the utility of single‐cell isolation to advance the centuries‐old problem of disentangling life stages and cryptic species. [ABSTRACT FROM AUTHOR]

Kiwifruit ripening and senescence after harvesting are closely related to its economic value. Transcriptome analysis and biochemical parameters were used to investigate the differences in gene expression levels and the potential regulation of cell wall metabolism in kiwifruit treated with ozone, thereby regulating fruit softening and prolonging postharvest life. Compared to the control group, the activities of the cell wall modification enzyme were lower under ozone treatment, the content of polysaccharide in the cell wall of primary pectin and cellulose was higher, and the content of soluble pectin was lower. Meanwhile, ozone treatment delayed the degradation of the cell wall mesosphere during storage. A total of 20 pectinesterase (PE)‐related genes were identified by sequencing analysis. The data analysis and quantitative polymerase chain reaction results confirmed that cell wall modifying enzyme genes played an important role in softening and senescence after harvesting, which may reduce or induce the expression of certain genes affecting cell wall metabolism. Ozone treatment not only regulates active genes such as xyloglucan endo glycosyltransferase/hydrolase, cellulose synthase, polygalacturonase, and PE to maintain the quality of fruit after harvest but also acts synergically with cell wall modifying enzymes to inhibit the degradation of cell wall, resulting in changes in the ultrastructure of cell wall, thereby reducing the hardness of kiwifruit. In addition, according to the results of cis‐acting elements, cell wall degradation is also related to downstream hormone signaling, especially PE‐related genes. These results provide a theoretical basis for studying the mechanism of firmness and cell wall metabolism difference of kiwifruit and also lay a good foundation for further research. [ABSTRACT FROM AUTHOR]

Diabetes is a worldwide leading cause of mortality and morbidity, and type 1 diabetes (T1D) is the most common chronic autoimmune disorder in children and young adults. There is currently no definitive treatment for T1D. Stem cell administration appeared as an experimental treatment showing an improvement in the clinical outcome and restoration of tissue architecture in many disorders. Mesenchymal stem cells (MSCs) showed their capacity to generate insulin-producing cells as a promising potential treatment for diabetic patients. The present work aimed to study the effect of MSCs on the ultrastructure and laboratory findings of streptozotocin (STZ)-induced T1D in rats. Sixty male albino rats were divided into three groups: group I (control), group II (STZ-induced diabetic), and group III (STZ-induced diabetic-treated with MSCs). Ultrastructure study, as well as glucose and insulin measurements, was done for the three groups. The diabetic group showed significantly higher glucose and lower insulin levels compared to the other two groups, while the diabetic group treated with MSCs showed a non-significant difference compared to the control group. On the ultrastructure level, the diabetic group showed nuclear degeneration, loss of cytoplasmic organelles, loss of cell membranes, pyknotic nuclei, and apoptosis. All these findings were mostly reversed by the use of MSCs in the diabetic-treated group. We conclude that the use of MSCs reversed the laboratory and ultrastructure effect of STZ-induced T1D in rats. [ABSTRACT FROM AUTHOR]

3D bioengineered skeletal muscle macrotissues are increasingly important for studies of cell biology and development of therapeutics. Tissues derived from immortalized cells obtained from patient samples, or from pluripotent stem cells, can be co-cultured with motor-neurons to create models of human neuromuscular junctions in culture. In this study, we present foundational work on 3D cultured muscle ultrastructure, with and without motor neurons, which is enabled by the development of a new co-culture platform. Our results show that tissues from Duchenne muscular dystrophy patients are poorly organized compared to tissues grown from healthy donor and that the presence of motor neurons invariably improves sarcomere organization. Electron micrographs show that in the presence of motor neurons, filament directionality, banding patterns, z-disc continuity, and the appearance of presumptive SSR and T-tubule profiles all improve in healthy, DMD-, and iPSC-derived muscle tissue. Further work to identify the underlying defects of DMD tissue disorganization and the mechanisms by which motor neurons support muscle are likely to yield potential new therapeutic approaches for treating patients suffering from Duchenne muscular dystrophy. [ABSTRACT FROM AUTHOR]

DYT1 dystonia is associated with decreased striatal dopamine release. In this study, we examined the possibility that ultrastructural changes of nigrostriatal dopamine terminals could contribute to this neurochemical imbalance using a serial block face/scanning electron microscope (SBF/SEM) and three‐dimensional reconstruction to analyse striatal tyrosine hydroxylase‐immunoreactive (TH‐IR) terminals and their synapses in a DYT1(ΔE) knockin (DYT1‐KI) mouse model of DYT1 dystonia. Furthermore, to study possible changes in vesicle packaging capacity of dopamine, we used transmission electron microscopy to assess the synaptic vesicle size in striatal dopamine terminals. Quantitative comparative analysis of 80 fully reconstructed TH‐IR terminals in the WT and DYT1‐KI mice indicate (1) no significant difference in the volume of TH‐IR terminals; (2) no major change in the proportion of axo‐spinous versus axo‐dendritic synapses; (3) no significant change in the post‐synaptic density (PSD) area of axo‐dendritic synapses, while the PSDs of axo‐spinous synapses were significantly smaller in DYT1‐KI mice; (4) no significant change in the contact area between TH‐IR terminals and dendritic shafts or spines, while the ratio of PSD area/contact area decreased significantly for both axo‐dendritic and axo‐spinous synapses in DYT1‐KI mice; (5) no significant difference in the mitochondria volume; and (6) no significant difference in the synaptic vesicle area between the two groups. Altogether, these findings suggest that abnormal morphometric changes of nigrostriatal dopamine terminals and their post‐synaptic targets are unlikely to be a major source of reduced striatal dopamine release in DYT1 dystonia. [ABSTRACT FROM AUTHOR]

In this study, we present the first integrative revision of the Boreal and Arctic calcareous sponges of the genus Leucosolenia with a specific focus on its biodiversity in the White Sea. The material for this work included a combination of newly collected specimens from different regions of the North-East Atlantic and the White Sea and historical museum collections. An integrative analysis was implemented based on vast morphological data (light microscopy, scanning and transmission electron microscopy), microbiome observations, ecological data, accompanied by molecular phylogenetic and species' delimitation analyses based on three nuclear markers (28S rRNA, 18S rRNA, and histone 3). We demonstrate that Leucosolenia complicata , previously reported from Arctic waters, is restricted to the North-East Atlantic, while in the Arctic, Leucosolenia diversity is represented by at least four species: Leucosolenia corallorrhiza , Leucosolenia variabilis , and two new species, one of which is described herein under the name Leucosolenia creepae sp. nov.. The molecular phylogeny analysis supports the species identity of these species. In addition to conventional morphological characters, new informative fine morphological characters (skeleton and oscular crown organization; cytological structure, including morphotypes of symbiotic bacteria) were found, providing a baseline for further revision of this group in other regions. [ABSTRACT FROM AUTHOR]

Encystment is a natural process that involves cyst formation, and at least some species of tardigrades can form cysts. However, the encystment process and cyst structure among tardigrades are still poorly understood. Despite some aspects of the encysted animals' systems organisation being examined in the past, the morphology and structure of the nervous system have never been thoroughly investigated. This study covers anatomical, histological and morphological details and proposes physiological aspects of the nervous system in encysted Thulinius ruffoi up to 11 months duration in encystment. This is the first record of the nervous system organisation in a species belonging to the family Doryphoribiidae. The cyst formation results in morphological changes in the nervous system. It comprises central and peripheral elements, which may be observable even after many months since the cyst formation. Based on the nervous system's organisation in cysts, there is no sign that histolysis is a part of encystment. [ABSTRACT FROM AUTHOR]

Rodents can present a large number of morphophysiological variations of the digestive system, allowing them to feed on different diets. In this context, the objective was to describe the morphology of the major salivary glands of Galea spixii. Gross dissection, light microscopy and transmission and scanning electron microscopy assessments of 12 specimens were carried out. The results demonstrate that the G. spixii has four pairs of major salivary glands (parotid, mandibular, zygomatic and sublingual). The glands are lobulated, composed acinar, consisting of mucous and serous acini, with the predominant parotid gland being serous and the zygomatic gland mucous. The mandibular and sublingual glands are mixed. The glands secrete acidic and neutral mucins and commonly present intercalary, striated and excretory ducts. The ultrastructure of the acinar cells suggests the presence of great cellular activity, with a cytoplasm taken by a rough endoplasmic reticulum, many mitochondria, vesicles of electrolucent nature and dispersed chromatin. In conclusion, the major salivary glands of G. spixii are constituted of mucous and serous acini, with serous acini predominant in the mandibular areas. The data also suggest that these rodents may adopt different types of diets, which may explain their adaptability to semi‐arid environments. [ABSTRACT FROM AUTHOR]

Calcium plays a crucial role in plant growth and development, yet little is known about its function in endodormancy regulation. Tree peony (Paeonia suffruticosa), characterized by compound buds and large flowers, is well-known for its ornamental and medicinal value. To break bud dormancy release is a prerequisite of flowering and forcing culture, particularly during the Spring Festival. In this study, the Ca2+ chelator EGTA and Ca2+ channel blocker LaCl3 were applied, resulting in a significant delay in budburst during both chilling- and gibberellin (GA)- induced dormancy release in a dosage-dependent manner. As expected, the retardation of bud break was recovered by the supplementation of 30 mM CaCl2, indicating a facilitating role of calcium in dormancy release. Accordingly, several calcium-sensor-encoding genes including Calmodulin (CaM) and Ca2+-dependent protein kinases (CDPKs) were significantly upregulated by prolonged chilling and exogenous GAs. Ultrastructure observations revealed a decline in starch grains and the reopening of transport corridors following prolonged chilling. Calcium deposits were abundant in the cell walls and intercellular spaces at the early dormant stage but were enriched in the cytosol and nucleus before dormancy release. Additionally, several genes associated with dormancy release, including EBB1, EBB3, SVP, GA20ox, RGL1, BG6, and BG9, were differentially expressed after calcium blocking and recovery treatments, indicating that calcium might partially modulate dormancy release through GA and ABA pathways. Our findings provide novel insights into the mechanism of dormancy release and offer potential benefits for improving and perfecting forcing culture technology in tree peonies. [ABSTRACT FROM AUTHOR]

Micromorphological visualization of plant surface peculiarities provides valuable characters for the precise identification of plant species. Invasive alien species, introduced outside their native range, pose significant ecological, and health challenges. This study focuses on micromorphological investigations of selected invasive plant species belonging to the families Amaranthaceae, Asteraceae, Moraceae, Crassulaceae, Cannabaceae, Fabaceae, Commelinaceae, and Oxalidaceae. The study employs scanning electron microscopy (SEM) to analyze the species micromorphology of pollen structure in depth to characterize the sculpturing patterns. Additionally, the study examines the pollen characteristics of these invasive plants, including shape, size, and fertility, along with exine sculpturing. The maximum polar diameter was observed for Senna tora (41.2 μm). The spines in Achillea fragrantissima were measured to have dimensions of approximately 1.91 μm in length and 2.11 μm in width. The findings shed light on the allergy‐causing potential of these invasive species, providing crucial information for accurate identification and effective management strategies to safeguard the indigenous flora of the region. The study contributes to the knowledge base for botanists, taxonomists, policy makers, climate experts, and biodiversity specialists. Research Highlights: Integrating SEM to analyze the pollen of invasive species.Unveiling pollen micromorphology of invasive species.Exploring the microscopic realm of invasive pollen to accurately identify the species. [ABSTRACT FROM AUTHOR]

Ultrastructural findings immediately after pulsed field ablation (PFA) of the myocardium have not been described. The purpose of this study was to elucidate ultrastructural characteristics and differences between microsecond PFA at the 1- and 4-hour timepoints after pulse delivery and to compare them to irrigated radiofrequency ablation (RFA) lesions. Healthy swine underwent endocardial PFA or RFA followed by necropsy. Discrete microsecond PFA and irrigated RFA lesions were created in the ventricles with a lattice tip ablation catheter. Lesions were delivered in a manner so as to allow sampling to occur 1 and 4 hours after ablation. All lesions were located at necropsy, and samples were carefully obtained from within the lesion core, lesion periphery, and adjacent healthy myocardium. Transmission electron microscopic assessment was performed after fixation using paraformaldehyde and glutaraldehyde. One hour after microsecond PFA delivery, myocytes were noted to be significantly and uniformly disrupted. Clustered, misaligned, swollen mitochondria coupled with degenerating nuclei and condensed chromatin were visualized. These findings progressed over the subsequent few hours with worsening edema. Similar changes were seen with RFA but reduced in severity. However, there was prominent extravasation of red blood cells with occlusion of capillaries that was not seen in PFA. At the lesion periphery, an abrupt change in the degree of myocyte damage was observed with PFA but not RFA. Transmission electron microscopy demonstrates evidence of widespread destruction of myocytes as early as an hour after PFA and corroborates known histologic features such as sparing of vessels and sharp lesion margins. [ABSTRACT FROM AUTHOR]

The using of explosive substances and devices can lead to brain injuries, the diagnosis of which requires the determination of primary biomarkers. Therefore, the aim was to establish and study histopathological and ultrastructural changes in the rats brain after exposure to an air blast wave. The study was carried out on 18 male Wistar rats, which were randomly divided into two groups: sham (n=9) and experimental (n=9). The animals of the experimental group were anesthetized with halothane and gently fixed in a horizontal position on the abdomen with the front part of the rat's muzzle at a distance of 5 cm from the device opening and subjected to an excess pressure of 26-36 kPa. Animals of both groups were decapitated, brains were removed, fixed, histopathological and ultrastructural analyzes were conducted using standard methods. Intergroup differences were assessed by Mann-Whitney U-test. Light microscopy revealed primary lesions in the form of small focal, multifocal hemorrhages, cerebral vessel ruptures and microscopic ruptures of the brain substance. In almost all brain samples, there is a significant saturation of the venous vessels with the presence of erythrocyte stasis. Violation of the blood-brain barrier, the presence of edema of the perivascular space, and petechial hemorrhages in the neuropil were registered ultrastructurally. Based on the study results, a morphological algorithm for assessing primary histostructural intracranial brain injuries and their consequences after exposure to an air shock wave was proposed. The injuries were found to be caused by the direct traumatic effect of the air shock wave. In the acute post-traumatic period, histopathological and ultrastructural changes in the brain can manifest as changes in neurons and in the blood-brain barrier and be accompanied by perivascular multifocal small-focal hemorrhages, neuropil ruptures, edema of pericellular and perivascular spaces, which together can be considered as biomarkers of primary traumatic changes after exposure to an air shock wave.

The main functions of the mammalian cecum include microbial digestion of soluble and insoluble carbohydrates and the absorption of significant amounts of water and electrolytes. The present study was carried out to investigate the gross anatomy and electron microscopy of the cecum in adult camels (Camelus dromedarius), which still needs to be clarified. The study was conducted using samples collected from camels slaughtered at an abattoir in Omdurman, Sudan. The camel cecum, which occupied the right part of the abdomen, was situated caudally to the abomasum and directed caudally to the pelvis. It was short and S-shaped and began largely at the ceco-colic junction at the level of the fifth lumbar vertebra and became smaller towards its apex. Morphometrically, camel cecum constituted 3.2% of the large intestine length, 1% of both the small intestine and large intestine length, and 0.10% of the long axis of the animal body. Ultra-structurally, the cecal mucosa showed several folds and was lined by a simple columnar epithelium with many intestinal cells (enterocytes) and goblet cells; two types of enterocytes were shown: type one contained a dark cytoplasm and type two had a light cytoplasm. In conclusion, the cecum in dromedary camels was generally similar in structure to other domestic animals; however, they have a shorter cecum compared to ruminants.

BACKGROUND: Warm-needling moxibustion can effectively treat knee osteoarthritis. Degeneration, injury and fracture of the anterior cruciate ligament can affect the local stability of the knee joint, and then induce the formation of knee osteoarthritis. Whether warm-needling moxibustion can repair the injured cruciate ligament and the mechanism of action are still unclear. OBJECTIVE: To observe the effects of warm-needling moxibustion on the morphology of the anterior cruciate ligament and the expression of insulin growth factor-1 and transforming growth factor-β in rabbits with knee osteoarthritis and to clarify the mechanism of anterior cruciate ligament repair by warmneedling moxibustion. METHODS: Thirty New Zealand rabbits were randomly divided into blank group, model group and warm-needling moxibustion group, with 10 rabbits in each group. Knee osteoarthritis model was established by plaster cast immobilization. The blank group was not intervened. Rabbits in the model group rabbits were fixed in a rabbit holder for 15 minutes every day. The warm-needling moxibustion group was treated with warm acupuncture, once a day, 7 days as a course of treatment, a total of two courses. After treatment, the imaging changes of the anterior cruciate ligament were observed by MRI and MRI grading statistics were performed. Morphological changes of the anterior cruciate ligament were observed by transmission electron microscope and hematoxylin-eosin staining. mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were detected by RT-PCR and western blot, respectively. RESULTS AND CONCLUSION: MRI examination: Compared with the blank control group, the anterior cruciate ligament in the model group was thickened, edematous, and partially torn, and the difference in grading statistics was statistically significant (P < 0.05). Compared with the model group, the anterior cruciate ligament in the warm-needling moxibustion group was slightly thickened, with mild edema and no tearing, and the difference in grading statistics was statistically significant (P < 0.05). General observation: In the model group, the surface of the anterior cruciate ligament was glossy and faded, with the edge being covered with flocculent periosteum and obvious tissue necrosis; in the warm-needling moxibustion group, the surface of the ligament was glossy, and the ligament was in a normal helical shape. Hematoxylin-eosin staining: In the model group, there was obvious tissue necrosis in the anterior cruciate ligament, a large number of new capillaries, loosely arranged fibroblasts and collagen fibers. In the warm-needling moxibustion group, there was a small amount of tissue necrosis and few new vessels in the anterior cruciate ligament, and the cells and collagen fibers were loosely and irregularly arranged. Transmission electron microscopy: In the model group, the fibers in the anterior cruciate ligament were arranged in a disordered way with uneven thickness and distribution, and there are more fibroblasts that were irregular in morphology; in the warm acupuncture group, the fibers were basically arranged longitudinally, with uneven thickness and distribution, and a small number of oval-shaped fibroblasts were observed. RT-PCR and western blot assay: mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were significantly decreased in the model group compared with the blank control group (P < 0.05), but significant increased after treatment with warm-needling moxibustion (P < 0.05). To conclude, warm-needling moxibustion can alleviate anterior cruciate ligament injury and regulate the expression of insulin growth factor-1 and transforming growth factor-β to treat knee osteoarthritis. [ABSTRACT FROM AUTHOR]

Global contraction of biodiversity pushed most members of Felidae into threatened or endangered list except the domestic cat (Felis catus) thence preferred as the best model for conservation studies. One of the emerging conservation strategies is vitrification of ovarian tissue which is field-friendly but not yet standardized. Thus, our main goal was to establish a suitable vitrification protocol for feline ovarian tissue in field condition. Feline ovarian tissue fragments were punched with biopsy punch (1.5 mm diameter) and divided into 4 groups. Group 1 was fresh control (Fr), while the other three were exposed to 3 vitrification protocols (VIT_CT, VIT_RT1 and VIT_RT2). VIT_CT involved two step equilibrations in solutions containing dimethyl sulfoxide (DMSO) and ethylene glycol (EG) for 10 min each at 4 °C. VIT_RT1 involved three step equilibration in solutions containing DMSO, EG, polyvinylpyrrolidone and sucrose for 14 min in total at room temperature, while in VIT_RT2 all conditions remained the same as in VIT_RT1 except equilibration timing which was reduced by half. After vitrification and warming, fragments were morphologically evaluated and then cultured for six days. Subsequently, follicular morphology, cellular proliferation (expression of Ki-67, MCM-7) and apoptosis (expression of caspase-3) were evaluated, and data obtained were analysed using generalised linear mixed model and chi square tests. Proportions of intact follicles were higher in Fr (P = 0.0001) and VIT_RT2 (P = 0.0383) in comparison to the other protocols both post warming and after the six-day culture. Generally, most follicles remained at primordial state which was confirmed by the low expression of Ki-67, MCM-7 markers. In conclusion, VIT_RT2 protocol, which has lower equilibration time at room temperature has proven superior thus recommended for vitrification of feline ovarian tissue. • Domestic cat ovarian tissue vitrification protocol as a model for endangered felids. • Manifold cryoprotectants in vitrification media produce better post-warm integrity. • Lower equilibration time at room temperature ensures better tissue morphology. [ABSTRACT FROM AUTHOR]

BACKGROUND: As a dominant breed pig in southwest China, the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines, but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE: To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS: Twenty adult female Yunnan small-ear pigs were equally randomized into two groups. In the autologous Achilles tendon group, the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft, while in the sham-operated group, a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament. General conditions of each pig were observed and recorded before and 12 months after surgery. Ligaments and grafts were taken for gross observation and MAS scoring. Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments. The staining and arrangement of type I and type III collagen were evaluated by immunohistochemistry. Transmission electron microscopy was used to observe the type, size, diameter, ratio, and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION: All animals had normal diet and activity, good wound healing, no obvious inflammatory reaction, no local purulent infection, and no significant changes in mental and urinary conditions compared with those before surgery. The reconstructed cruciate ligament of the knee was intact, with no stiffness and normal range of motion. Both the anterior drawer and Lachman tests were negative. Gross observation of the graft: 12 months after surgery, the grafts was in good position, with good integrity, obvious tension, ligament color close to the original anterior cruciate ligament, and complete surface synovial coverage. Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS II type. In the sham-operated group, the intraarticular ligament was defined as MAS I type. Hematoxylin-eosin staining indicated that, 12 months after surgery, collagen fibers in the autologous Achilles tendon group began to appear bundled, isotropic, and uniformly arranged, with more obvious isotropic corrugations, and the nuclei were mainly linear or spindle-shaped, which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group. Immunohistochemistry results indicated that, 12 months after surgery, there was a higher expression of type I collagen and significantly less expression of type III collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group. The degree of type I and type III staining was similar in the two groups. Under the transmission electron microscope, the diameter, arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery, indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery. Through a comprehensive evaluation of animal general conditions, ligament general view, MAS score, hematoxylin-eosin staining, immunohistochemistry, and transmission electron microscopy observation, we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs, with good morphological, histological and ultrastructural results. [ABSTRACT FROM AUTHOR]

Background. The aim of the research was to determine the ultrastructure of the cells of the muscle wall of the mice gallbladder under the influence of ursodeoxycholic acid. Materials and methods. The animals were divided into 2 groups: experimental (n = 17) — mice which received ursodeoxycholic acid at a dose of 100.0 mg/kg and control (n = 13) — intact ones (distilled water). Histological and ultrastructural analyses of gallbladder wall samples of mice were performed. Results. Mitotic figures of myocytes in the wall of the gallbladder bottom (1.794 ± 0.103 %) and body (1.607 ± 0.095 %) in the experimental group of mice were significantly more frequent compared to the controls (0.946 ± 0.058 % and 0.873 ± 0.061 %) (p

Abstract Background Nanoparticle-based drugs are new inventions in the management of the Human immunodeficiency virus (HIV) pandemic, especially resistant forms of the virus in anatomical sanctuary sites and organs such as the testis. However, safety issues must be resolved to attain the optimal potential of newer nano-drug formulations. Aim The study investigated the toxicological potential of synthesized Tenofovir Nanoparticles (TDF-N) on testicular indices when used for the prevention and treatment of HIV. Methodology Fifteen male Sprague–Dawley (SD) rats with weight ranging from 230 g to 250 g were randomly assigned into groups A (control, saline), B (TDF), and C (TDF-N). The testes were removed for sperm analysis and processed for H/E and PAS stains. Cell counts and cellular measurements; the diameter and the area of the testicular seminiferous tubules were measured using ImageJ and Leica software 2.0. Results A significant reduction (p

Abstract Increasing temperature influences the habitats of various organisms, including microscopic invertebrates. To gain insight into temperature-dependent changes in tardigrades, we isolated storage cells exposed to various temperatures and conducted biochemical and ultrastructural analysis in active and tun-state Paramacrobiotus experimentalis Kaczmarek, Mioduchowska, Poprawa, & Roszkowska, 2020. The abundance of heat shock proteins (HSPs) and ultrastructure of the storage cells were examined at different temperatures (20 °C, 30 °C, 35 °C, 37 °C, 40 °C, and 42 °C) in storage cells isolated from active specimens of Pam. experimentalis. In the active animals, upon increase in external temperature, we observed an increase in the levels of HSPs (HSP27, HSP60, and HSP70). Furthermore, the number of ultrastructural changes in storage cells increased with increasing temperature. Cellular organelles, such as mitochondria and the rough endoplasmic reticulum, gradually degenerated. At 42 °C, cell death occurred by necrosis. Apart from the higher electron density of the karyoplasm and the accumulation of electron-dense material in some mitochondria (at 42 °C), almost no changes were observed in the ultrastructure of tun storage cells exposed to different temperatures. We concluded that desiccated (tun-state) are resistant to high temperatures, but not active tardigrades (survival rates of tuns after 24 h of rehydration: 93.3% at 20 °C, 60.0% at 35 °C, 33.3% at 37 °C, 33.3% at 40 °C, and 20.0% at 42 °C).

Cephalopods, which have a rapid growth rate and short life cycle, are regarded as an important marine fishing resource. Sepioteuthis lessoniana is among the most widely distributed species in the Loliginidae family in the Indo-Western Pacific Ocean. Its meat is delicious and nutritious. It is widespread in the East China Sea, South China Sea, and other marine areas in China, and it is considered an important local marine resource. In this study, the ultrastructure of the oviducal gland of S. lessoniana was investigated for the first time via anatomical dissection, tissue sectioning, and electron microscope projection. The external morphology and internal structure of the gland were clearly described, as well as the role of the oviduct gland in the reproductive activities of cephalopods. In this experiment, all the samples were collected from the open ocean, with the samples of wild S. lessoniana coming from marine areas in Fujian Province. The body surfaces of the samples were healthy and undamaged, with an average mantle length of (17.5±6.4) cm and an average body weight of (392.0±76.0) g. The samples were dissected using standard anatomical methods, and the glandular characteristics were recorded. The oviducal gland was dissected with a scalpel for tissue sectioning and preserved for electron microscope projection and observation. The experimental results showed that S. lessoniana had a single oviducal gland, which was located on the right side of its abdominal cavity. The sexually mature gland was milky white overall, with brownish-yellow pigmentation near the inner shell. The oviducal gland was enveloped in a transparent membrane, and regular gaps were visible. The gland consisted of three parts: the proximal oviducal gland, the distal oviducal gland, and the transparent valve. The proximal oviducal gland was infundibular and connected with the hyaline oviduct. The distal oviducal gland was cylindrically tapering, and a single mature egg was observed inside. The transparent valve was at the end of the gland, also known as the valve. The microstructure of the oviducal gland was observed via a microscope. The gland was composed of a glandular wall, lamellar, and muscle tissue. The glandular wall tissue was composed of adventitia, loose connective tissue, and a small amount of muscle tissue, blood vessels, and ducts that were scattered in the connective tissue. The transparent valve was composed of columnar epithelial cells and muscle tissue. A small number of water-droplet goblet cells were dispersed between the epithelial cells, while a large number of cilia were generated on the exterior. The lamellar was attached to the glandular wall tissue and distributed in layers within the gland; it was mainly composed of ciliated columnar epithelium and support cells. When the oviducal gland was at various development stages, the cell types of the lamellar and the size of the intercellular space within the leaflet were different. When the gland was immature, the lobe had a regular shape, with more connective tissue in the center and a single layer of columnar epithelial cells on each side. When the gland was about to mature, the connective tissue increased, the number of columnar epithelial cells decreased, and a large number of mucous acinus were simultaneously generated. After the glands matured and spawned, the amount of connective tissue decreased, the columnar epithelial cells disappeared, the mucous acinus ruptured, and the secretory leaflets were filled with secretory substances. The individual oviduct gland secretory cells of S. lessoniana were large and contained many closely arranged mucus granules, which were round or oval in shape. The cytoplasm contained many organelles, such as the endoplasmic reticulum, mitochondria, and Golgi apparatus. In addition, secretory cells were continuously distributed outside the cell with cilia and numerous secretory granules. The nucleus of the muscle cells in the hyaline valve was irregular, and a substantial number of myofilaments, collagen fibers, and capillaries could be seen around it. The oviducal gland is an important gland in the reproductive system of female cephalopods. The morphological characteristics of oviducal glands, such as the number, shape, size, and presence or absence of pigmentation, are some of the distinguishing characteristics between different cephalopod species. The tissue structure and cell types of the cephalopod oviducal glands changed with the growth of individuals. The changes in oviducal glands in S. lessoniana were similar to those in Loligo forbesi. The secretory lobes successively generated an increasing number of mucous acini and secretory substances as cell types shifted. These secreted substances had multiple functions, such as attracting sperm during the sperm-egg hatching process, expanding the chorionic membrane, forming the vitelline space, and regulating the osmotic pressure between the fertilized egg and the ambient seawater. However, the oviducal glands of the orders Cuttlefish and Liliformes differ from those of the order Occarpus, whose oviducal glands had the function of storing sperm. It is concluded that the oviducal gland primarily plays a secretory function in the reproductive activities of cephalopods and that its secretory material forms the second layer of the egg membrane of fertilized eggs, ensuring the normal hatching of fertilized eggs.

The endemic chub Squalius tenellus (Heckel, 1843) was introduced more than 100 years ago to Lake Blidinje (Bosnia-Herzegovina). Only 1 species of enteric helminth was found in a sample of 35 chubs, the tapeworm Caryophyllaeus brachycollis (Janiszewska, 1953). The paper includes histopathological investigation with identification of innate immune cells involved in host reaction and molecular data allowed correct designation of the cestode species. Of 35 specimens of chub examined, 21 (60%) harboured individuals of C. brachycollis and a total of 1619 tapeworms were counted, the intensity of infection ranged from 1 to 390 worms per fish (46.2 ± 15.3, mean ± s.e.). Histopathological and ultrastructural investigations showed strict contact between the worm's body and the epithelia and increase in the number of mucous cells, rodlet cells among the epithelial cells. Within the tunica propria-submucosa, beneath the site of scolex attachment, numerous neutrophils and mast cells were noticed. This is the first study of the occurrence of C. brachycollis in chub from Lake Blidinje and on the response of the innate immune cells of S. tenellus to this tapeworm. Interestingly, in 3 very heavily infected chubs, perforation of the intestinal wall was documented; this is uncommon among cestodes which use fish as a definitive host.

AbstractG. mellonella is a promising species for use in the biodegradation of plastics. It is easy to breed and has high resistance to diverse climatic conditions, which is particularly valuable when considering its potential application in the decomposition of plastics. Thus, it demonstrated the capacity for biodegradation of the most common types of plastics, such as polyethylene (PE) and polypropylene (PP). However, reports on whether consumed plastics or their decomposition products will adversely affect the structure and functioning of the internal organs are rather poor. The studies aimed to determine whether the consumption of PP by a greater wax moth larvae caused any ultrastructural changes in the organs of the animal’s body, evaluate the survival rate of the animals, and describe their reproduction. Thus, this study provided an understanding of histological and ultrastructural changes caused, or not caused, by the PP diet. We investigated three organs – midgut, silk gland, and fat body – under PP consumption by G. mellonella caterpillars (7th instar larvae). The level of reactive oxygen species (ROS) in selected organs, as well as the ability of larvae to survive and undergo metamorphosis were also examined. The animals were divided into four groups: G0-C, G0-S, G0-24, and G0-48. The research used transmission electron microscopy (TEM), confocal microscopy, and flow cytometry. Our study showed that a diet containing PP did not affect internal organs at the ultrastructural level. Cells in the analyzed organs – midgut, silk gland, and fat body – showed no degenerative changes. An increase in the intensity of autophagy and cell vacuolization was noted, but they probably act as a survival pathway. These observations suggest that the final larval stage of the greater wax moth can potentially be applied in PP biodegradation.

Abstract Background The most frequent kind of dementia in the senior population is Alzheimer's disease (AD). Antioxidant quercetin has a low bioavailability. The bioavailability of quercetin nanoparticles was demonstrated to be higher. Acrolein is thought to be the strongest unsaturated aldehyde. Acrolein contributes to the propagation of oxidative damage and thus the aetiology of AD. This study aimed to investigate histopathological and ultrastructural changes that may arise in the hippocampus following acrolein treatment. Quercetin nanoparticles' ameliorative and protective effects on acrolein-induced neurotoxicity and oxidative stress were assessed. Results We successfully synthesised quercetin nanoparticles with uniform size distributions and particle diameters in the range of 3.63–4.57 nm using transmission electron microscopy (TEM) and 3.7 nm using dynamic light scattering (DLS). Administration of acrolein was associated with histopathological alterations in the hippocampal structure, such as increased apoptotic neurones, dystrophic changes, neuronophagia, and atrophic ischaemia in cells, as well as marked damage to the ultrastructure of the hippocampus, which was obvious in shrunken pyramidal neurones with pyknotic nuclei and completely degenerated chromatin material, as well as in damaged astrocytes and microglial cells. Treatment with quercetin nanoparticles has been found to protect against and ameliorate the toxic effects and oxidative stress induced by acrolein in the hippocampus. Conclusions This could pave the way for additional research in nanomedicine and a new line of therapeutic intervention in AD using nanoparticles such as quercetin nanoparticles.

Background: Nanoparticle-based drugs are new inventions in the management of the Human immunodeficiency virus (HIV) pandemic, especially resistant forms of the virus in anatomical sanctuary sites and organs such as the testis. However, safety issues must be resolved to attain the optimal potential of newer nano-drug formulations. Aim: The study investigated the toxicological potential of synthesized Tenofovir Nanoparticles (TDF-N) on testicular indices when used for the prevention and treatment of HIV. Methodology: Fifteen male Sprague–Dawley (SD) rats with weight ranging from 230 g to 250 g were randomly assigned into groups A (control, saline), B (TDF), and C (TDF-N). The testes were removed for sperm analysis and processed for H/E and PAS stains. Cell counts and cellular measurements; the diameter and the area of the testicular seminiferous tubules were measured using ImageJ and Leica software 2.0. Results: A significant reduction (p < 0.05) in sperm count was noticed in the TDF-N group. Also observed in the TDF and TDF-N groups was a significant reduction (p < 0.05) in sperm motility and in the number of dead sperms compared with the control. Sperm abnormalities such as distorted basement membranes, loss of germ cells, hypocellular interstitium, and loss of spermatogenic series were increased in the TDF and TDF-N groups. There was also a significant reduction (p < 0.05) in the cell count, diameter, and area of seminiferous tubules observed in these groups. Conclusion: TDF and TDF-N may be detrimental to the testis and testicular tissue, leading to significantly reduced sperm counts, motility, and ultimately–male fertility. [ABSTRACT FROM AUTHOR]

The skin color of the large yellow croaker (Larimichthys crocea) is a crucial indicator to determine its economic value. However, the location of pigment cells in the skin structure is uncertain. To determine the pigment cell type in the skin, the vertical order and ultrastructure of pigment cells were examined using light microscopy and transmission electron microscopy. Both dorsal and ventral skins comprise the epidermis, dermis, and hypodermis. Xanthophores, melanophores, and iridophores were observed in the dermis of the dorsal skin, whereas the latter two were in the dermis of the ventral skin. Interestingly, the size of xanthophores in the dorsal skin was significantly smaller than that of xanthophores in the ventral skin; however, the density of dorsal xanthophores was significantly higher than that of ventral xanthophores. The type L‐iridophores with large crystalline structures were observed in the uppermost area of the upper pigment layer, which contributed to the strikingly metallic luster shown by the ventral skin. The melanophores were exclusively found in the dorsal skin, offering the purpose of camouflage. Taken together, our results indicated that the pigment cells display different arrangement patterns between dorsal and ventral skin, and the golden color in the ventral skin results from the coexistence of light‐reflecting iridophores and light‐absorbing xanthophores. [ABSTRACT FROM AUTHOR]

Cannabaceae species garner attention in plant research due to their diverse secretory structures and pharmacological potential associated with the production of secondary metabolites. This study aims to update our understanding of the secretory system in Hops (Humulus lupulus L.), an economically important species especially known for its usage in beer production. For that, stems, leaves, roots, and inflorescences were collected and processed for external morphology, anatomical, histochemical, ultrastructural and cytochemical analyses of the secretory sites. Our findings reveal three types of secretory structures comprising the secretory machinery of Hops: laticifer, phenolic idioblasts and glandular trichomes. The laticifer system is articulated, anastomosing and unbranched, traversing all plant organs, except the roots. Phenolic idioblasts are widely dispersed throughout the leaves, roots and floral parts of the species. Glandular trichomes appear as two distinct morphological types: capitate (spherical head) and peltate (radial head) and are found mainly in foliar and floral parts. The often-mixed chemical composition in the secretory sites serves to shield the plant from excessive UVB radiation, elevated temperatures, and damage inflicted by herbivorous animals or pathogenic microorganisms. Besides the exudate from peltate glandular trichomes (lupulin glands), latex and idioblast content are also likely contributors to the pharmacological properties of different Hop varieties, given their extensive presence in the plant body. [ABSTRACT FROM AUTHOR]

Camellia sinensis L. is an important crop in Indonesia as healthy beverage that contains several secondary metabolism compounds, such as polyphenols and catechins. Tissue culture including somatic embryogenesis and organogenesis has been used for propagating plant for various needs. In this present shortcommunication, scanning electron microscopic (SEM) analysis of tea was conducted and discussed. This study aimed to investigate surface ultrastructure of TRI2025 embryo axis tea clone cultured on Murashige and Skoog (MS) medium containing 2,4-Dichlorophenoxyacetic acid (2,4-D). The results revealed two different forms of explant's development, i.e. somatic embryo and transitional form between somatic embryogenesis and organogenesis; or called by "Globular-like Structure" (GLS). Surface ultrastructure analys is of somatic embryo and GLS revealed respect ively many stages of somatic embryo development i.e. globular, torpedo, and cotyledon stage, and leaf development form GLS regeneration. [ABSTRACT FROM AUTHOR]

The ascomycete fungus Botryosphaeria dothidea is the causal agent of postharvest rot in kiwifruit. Production and spreading of conidia, differentiation of infection hyphae and invasion of intact fruit surfaces are required to cause fruit rot disease. In several pathogenic fungi, the homologue of the Saccharomyces cerevisiae mitogen activated protein kinase (MAPK) gene Fus3 has been identified as a pathogenicity factor. To investigate the role of the Fus3 homologue in B. dothidea, the gene was deleted and the resulting ∆fus3 mutants were characterized. On artificial media, strong growth defects of ∆fus3 strains were observed, mycelia were non‐pigmented, and mutants were unable to conidiate. Scanning electron microscopy showed that the wild‐type strain differentiated thick hyphae on the surface of kiwifruits, and nose‐down growth of hyphae indicated tight attachment to the fruit. In contrast, hyphae of ∆fus3 strains mutants were thinner and did not firmly attach to the fruit surface. Importantly, pathogenicity test revealed that ∆fus3 strains were unable to cause disease symptoms on intact kiwifruits. These results shown here identify Fus3 of B. dothidea as a novel pathogenicity factor of the ripe rot pathogen on kiwifruit. [ABSTRACT FROM AUTHOR]

The scaffolding protein programmed cell death protein 10 (Pdcd10) has been demonstrated to play a critical role in renal epithelial cell homeostasis and function by maintaining appropriate water reabsorption in collecting ducts. Both ureter and kidney collecting duct systems are derived from the ureter bud during development. Here, we report that cadherin-16 (Cdh16)- cre drives gene recombination with high specificity in the ureter, but not the bladder, urothelium. The consequences of Pdcd10 deletion on the stratified ureter urothelium were investigated using an integrated approach including messenger RNA (mRNA) expression analysis, immunocytochemistry, and high-resolution confocal and electron microscopy. Loss of Pdcd10 in the ureter urothelium resulted in increased expression of uroplakins (Upks) and keratins (Krts), as well as hypertrophy of the ureter urothelium with an associated increase in the number of proliferation marker protein Ki-67 (Ki67)-expressing cells specifically within the basal urothelium layer. Ultrastructural analysis documented significant modification of the intracellular membrane system, including intracellular vesicle genesis and transport along the basal- to umbrella-cell-layer axis. Additionally, Pdcd10 loss resulted in swelling of Golgi compartments, disruption of mitochondrial cristae structure, and increased lysosomal fusion. Lack of Pdcd10 also resulted in decreased fusiform vesicle formation in umbrella cells, increased secretion of exosome vesicles, and alteration in microvillar structure on apical membranes. Our findings indicate that Pdcd10 expression and its influence on homeostasis is associated with modulation of endomembrane trafficking and organelle biogenesis in the ureter urothelium. [ABSTRACT FROM AUTHOR]

This study aims to investigate the effects of melatonin on follicular growth, viability and ultrastructure, as well as on the levels of mRNA for antioxidant enzymes, reactive oxygen species (ROS) and meiotic progression in oocytes from in vitro cultured bovine early antral follicles. To this end, isolated early antral follicles (500–600 μm) were cultured in TCM‐199+ alone or supplemented with 10−6, 10−7 or 10−8 M melatonin at 38.5°C with 5% CO2 for 8 days. Follicle diameters were evaluated at days 0, 4 and 8 of culture. At the end of culture, ultrastructure, chromatin configuration, viability (calcein‐AM and ethidium homodimer‐1 staining), and the levels of ROS and mRNA for catalase (CAT), superoxide dismutase (SOD) and peroxiredoxin 6 (PRDX6) and glutathione peroxidase (GPx) were investigated in oocyte‐granulosa cell complexes (OGCs). The results showed that early antral follicles cultured with 10−6 and 10−8 M melatonin had a progressive and significant increase in their diameters throughout the culture period (p <.05). Additionally, oocytes from follicles cultured with 10−7 or 10−8 M melatonin had increased fluorescence for calcein‐AM, while those cultured with 10−6 or 10−7 M had reduced fluorescence for ethidium homodimer‐1. Different from follicles cultured in other treatments, those cultured with 10−8 M melatonin had well‐preserved ultrastructure of oocyte and granulosa cells. Melatonin, however, did not influence the levels of ROS, the mitochondrial activity, oocyte meiotic resumption and expression mRNA for SOD, CAT, GPX1 and PRDX6. In conclusion, the presence of 10−8 M melatonin in culture medium improves viability and preserves the ultrastructure of oocyte and granulosa cells of early antral follicles cultured in vitro. [ABSTRACT FROM AUTHOR]

Endoparasitic annelids living inside another annelid host are known, particularly with regard to Oenonidae, but in general are poorly studied. The dorvilleid Veneriserva pygoclava is known from southern California, and its genus name (Latin = Venus's servant) alludes to the close association with the host aphroditid scaleworm Aphrodita longipalpa. Little is known on fundamental questions on the biology of Veneriserva pygoclava. What is its mode of reproduction? How do they feed? How do they penetrate the host? We have studied multiple parasitized hosts and V. pygoclava specimens, using an integrative approach, combining µCT, histology, and electron microscopy. 3D reconstructions from µCT data of a parasitized Aphrodita show the exact position of the parasites in their natural condition within the host's coelomic cavity. Ultrastructural investigations of the parasites revealed interesting adaptations to their lifestyle such as the complete reduction of their gut, despite the presence of a functional jaw apparatus and a modified epidermis enabling nutrient uptake from the host's coelomic fluid. In addition to these, we also investigated spermatogenesis and oogenesis in V. pygoclava. Sperm morphology indicates an external fertilization of eggs within the coelomic cavity of the host. Mature male and female parasites living inside the same mature host and the presence of juvenile V. pygoclava within juveniles of Aphrodita suggest an obligate form of parasitism with a very early penetration of the hosts. In addition to our detailed morphological investigation, we conducted a phylogenetic analysis showing the position of Veneriserva within Dorvilleidae and its position was recovered nested among taxa of the Iphitime. Our phylogenetic analyses also show that the taxation Ophryotrocha puerilis siberti should be given full species rank and referred to as Ophryotrocha siberti. Finally, we publish here the full mitochondrial genome of V. pygoclava and discuss its novel gene order with reference to other annelids. [ABSTRACT FROM AUTHOR]