Search Results (36)

Mast cells (MCs) are immune cells that produce numerous immunological mediators involved in inflammatory and allergic responses. Increased numbers of MCs are observed in chronic inflammatory reactions in organs such as the colon. There, MCs seem to participate in deleterious immune responses and tissue damage, but the detailed mechanisms of their activation are not known, mostly because procedures to obtain MC primary cultures from the colonic mucosa are expensive and time-consuming and present low yield. Here we describe a protocol to obtain MCs from the colonic mucosa (cmMCs) of C57BL/6 mice with high yield, viability and purity. Mucosal colon cells were dispersed by enzymatic digestion, and cmMCs were isolated by Percoll continuous-gradient centrifugation. This method allowed for the purification of 1,446,667 ± 112,442 cell/g of mucosal tissue, with 87.22% viability and 95.16% purity. The mucosal-like phenotype was predominant in isolated cmMCs, characterized by weak toluidine blue staining but strong expression of MC protease-1 (Mcpt1). Activation assays showed that freshly isolated cmMCs increased intracellular calcium and showed degranulation in response to ATP or IgE-antigen-dependent FcεRI cross-linking. This highly reproducible technique is cost-effective and requires no specialized equipment. This protocol could be applied in research related to inflammatory bowel disease, colon cancer or food allergies. Full article

The second stomach compartment (C2) of the dromedary camel (Camelus dromedarius) plays an important role in digestion. However, detailed morphological and histochemical data remain limited. This study aimed to investigate the gross anatomy, histological organization, histometric features, and histochemical distribution of muco-substances in C2. The study was conducted on twenty dromedary camels, including fetuses and adults. Gross anatomical observations were performed on eight fresh and fixed specimens, while histological, histometric, and histochemical analyses were carried out on samples from twelve adult camels using routine and special staining techniques to identify neutral and acidic mucins. C2 was the smallest gastric compartment, located on the right side of the abdominal cavity and partially continuous with C1. Its mucosa formed chambered zones supported by prominent longitudinal muscular bands. Histologically, C2 comprised glandular and non-glandular regions. The glandular mucosa contained gastric pits and branched tubular glands with mucous, chief, and parietal cells, whereas the non-glandular region was lined by keratinized stratified squamous epithelium. Submucosal lymphoid aggregations were observed near the C2–C3 junction. Histometric analysis revealed a markedly developed tunica muscularis. Strong PAS and Alcian blue reactions indicated abundant neutral and acidic mucins. These findings demonstrate that C2 is a structurally specialized compartment supporting digestion, mucosal immune defense, and adaptation to arid environments, clearly distinguishing it from the reticulum of true ruminants. Full article

Background/Objectives: Flea beetles (Coleoptera, Chrysomelidae: Alticinae) show extensive karyotypic diversity, yet cytogenetic and genomic data remain scarce for many taxa. Species of the genus Podagrica are characterized by unusually high chromosome numbers compared with the modal condition in Alticinae, suggesting a history of chromosomal fissions. This study aimed to characterize the karyotype and repetitive DNA composition of Podagrica fuscicornis, with special emphasis on the satellitome and its contribution to chromosome organization. Methods: Male specimens of P. fuscicornis collected in southern Spain were analyzed using conventional cytogenetic techniques, including Giemsa staining, DAPI staining, and C-banding. Fluorescence in situ hybridization was employed to map nucleolar organizer regions (NORs), telomeric repeats, and major satellite DNA (satDNA) families. The satellitome was characterized using Illumina short-read sequencing and analyzed with the RepeatExplorer2/TAREAN pipeline to identify satDNA families and estimate their genomic abundance and divergence. Results: The male karyotype of P. fuscicornis was 2n = 40 (38 + XY), with an Xy_p sex chromosome system. Constitutive heterochromatin was mainly pericentromeric, and the Y chromosome was largely heterochromatic. NORs were located on a single autosomal pair, and the ancestral insect telomeric motif (TTAGG)_n was detected at chromosome ends. The satellitome comprised at least 70 different satDNA families, representing 9.51% of the genome, some of them related to transposable elements. Ten of these 70 satDNAs are shared in other Alticinae species. The most abundant families were primarily localized in pericentromeric regions and showed differential distribution between autosomes and sex chromosomes. Conclusions: These results indicate that extensive chromosomal fissions and high satDNA dynamics could drive the high chromosome number and heterogeneous genome organization in P. fuscicornis, highlighting the role of repetitive DNA in karyotype evolution within Chrysomelidae. Full article

Background: Pathology education requires innovative experimental teaching approaches to enhance clinical competency. This study evaluated the integration of special staining techniques into pathology curricula to improve diagnostic confidence and practical skills. Methods: The reform involved 227 medical students, incorporating acid-fast, PAS, GMS, Congo red, and other special stains into laboratory sessions. Diagnostic confidence was surveyed, and theoretical and practical exam scores were compared with 180 students from a previous grade. Statistical analysis was performed using GraphPad Prism 7.0. Results: Practical exam scores significantly improved (86.0 ± 17.2 vs. 82.2 ± 18.9, p < 0.001), while theoretical scores remained unchanged. Diagnostic confidence strongly correlated with morphological recognition, particularly for acid-fast and fungal stains. Student feedback noted challenges such as staining artifacts. Conclusion: Integrating special staining enhances practical skills and diagnostic confidence, effectively bridging basic and clinical training. Expanding such modules is recommended to advance competency-based medical education. Full article

The pervasive environmental contamination by micro- and nanoplastics (MNPs) presents a formidable analytical challenge, necessitating the development of rapid and sensitive detection methods. While conventional techniques often suffer from limitations in sensitivity and throughput, fluorescent probe-based technology has emerged as a powerful alternative. This review charts the evolution of these probes, from initial stains relying on hydrophobic adsorption to advanced molecular designs engineered for specific chemical recognition. We critically examine key operational mechanisms, including the solvatochromic response of Nile Red, polarity-discriminatory probes enabling a “microplastic rainbow,” and targeted systems achieving turn-on fluorescence via restriction of intramolecular rotation. Furthermore, we highlight cutting-edge signal enhancement strategies, such as plasmon- and metal-enhanced fluorescence, which amplify detection to the femtogram level. Special emphasis is placed on the distinct challenges posed by nanoplastics, including their propensity for aggregation in aqueous matrices that exacerbates false positives and their superior ability to breach biological barriers, and how AIE luminogens and PEF/MEF strategies mitigate these issues through enhanced signal-to-noise ratios and subcellular resolution, differing from their application to microplastics. Critically, we address the imperative for low-toxicity probe designs, emphasizing biocompatibility and biodegradability criteria to facilitate safe, long-term in vivo tracking and widespread ecological surveillance. The integration of these sophisticated probes with smart, “activate-on-target” systems is paving the way for next-generation MNP analysis, offering critical insights for environmental monitoring and toxicological assessment. Full article

This study aimed to determine whether the fetuses experienced hypoxic distress or intra-partum death due to compromised oxygen supply by quantitatively analyzing the lung structures of deceased captive South China Tiger (Panthera tigris amoyensis, SCT) cubs. Furthermore, it investigated the etiology and mechanisms of asphyxia in the deceased cubs and explored potential factors contributing to the high mortality rate of SCT cubs. The research focused on three deceased SCT cubs, using three deceased neonatal Amur Tigers (P. t. altaica, AT) with no inbreeding background as controls. Histological analysis of the cubs’ lungs was conducted using hematoxylin–eosin (H&E) and special staining techniques. The possible causes of death in SCT cubs were evaluated by comparing pulmonary pathological changes and morphological parameters, including alveolar space (P(ASP)) values, mean alveolar linear intercept (L_m), and mean thickness of alveolar septa (Tas). Our research reveals that hypoxia is one of the most significant factors contributing to the mortality of tiger cubs. Additionally, we hypothesize that inbreeding depression may lead to abnormal lung development (e.g., thickening of lung tissue) in SCT cubs. This study lays the foundation for comprehensive investigations into the mortality of highly inbred SCT cubs. Full article

The crystallization of soluble salts is one of the most significant agents of deterioration affecting porous building materials in historical architecture. This process not only compromises the physical integrity of the materials but also results in considerable aesthetic, structural, and economic consequences. Soluble salts involved in these processes may originate from geogenic sources—including soil leachate, marine aerosols, and the natural weathering of parent rocks—or from anthropogenic factors such as air pollution, wastewater infiltration, and the use of incompatible restoration materials. This study examines the role of capillary rise as a primary mechanism responsible for the vertical migration of saline solutions from the soil profile into historic masonry structures, especially those constructed with calcareous stones. It describes how water retained or sustained within the soil matrix ascends via capillarity, carrying dissolved salts that eventually crystallize within the pore network of the stone. This phenomenon leads to a variety of damage types, ranging from superficial staining and efflorescence to more severe forms such as subflorescence, microfracturing, and progressive mass loss. By adopting a multidisciplinary approach that integrates concepts and methods from soil physics, hydrology, petrophysics, and conservation science, this paper examines the mechanisms that govern saline water movement, salt precipitation patterns, and their cumulative effects on stone durability. It highlights the influence of key variables such as soil texture and structure, matric potential, hydraulic conductivity, climatic conditions, and stone porosity on the severity and progression of deterioration. This paper also addresses regional considerations by focusing on the context of Spain, which holds one of the highest concentrations of World Heritage Sites globally and where many monuments are constructed from vulnerable calcareous materials such as fossiliferous calcarenites and marly limestones. Special attention is given to the types of salts most commonly encountered in Spanish soils—particularly chlorides and sulfates—and their thermodynamic behavior under fluctuating environmental conditions. Ultimately, this study underscores the pressing need for integrated, preventive conservation strategies. These include the implementation of drainage systems, capillary barriers, and the use of compatible materials in restoration, as well as the application of non-destructive diagnostic techniques such as electrical resistivity tomography and hyperspectral imaging. Understanding the interplay between soil moisture dynamics, salt crystallization, and material degradation is essential for safeguarding the cultural and structural value of historic buildings in the face of ongoing environmental challenges and climate variability. Full article

Cartilage is a crucial component of the vertebrate skeletal system, providing structural integrity, flexibility, and adaptive functions across species. In teleost fish, cartilage exhibits significant morphological and functional diversity, providing specialized biomechanical properties essential for aquatic life. This study presents a detailed histological, immunohistochemical, and ultrastructural investigation of cartilage in molly fish (Poecilia sphenops), identifying five distinct types of cartilage: hyaline-cell, scleral, cell-rich hyaline, elastic cell-rich, and matrix-rich hyaline cartilage. Histological staining techniques revealed notable differences in cellular architecture and composition of the extracellular matrix among the cartilage types. Immunohistochemical analysis demonstrated the expression of S100 protein and acetylcholinesterase (Ach), suggesting their involvement in cartilage regulation and maintenance. Endochondral ossification was observed in the head and gill arches. Electron microscopy provided detailed insights into chondrocyte morphology, interactions between cartilage and the perichondrium, and interactions between telocytes and fibroblasts. The findings enhance our understanding of skeletal adaptations in teleost fish, emphasizing the functional diversity of cartilage in aquatic environments. This study contributes to evolutionary biology and may have implications for regenerative medicine and biomaterials research. Full article

Bryophytes, or non-vascular plants, provide valuable models for studying plant adaptation to land, as their physiology differs significantly from that of vascular plants. This study examines the cell wall structure of bryophytes, focusing on the tissue-specific distribution of cell wall epitopes in Sphagnum compactum (a peat moss) and Marchantia polymorpha (the model liverwort) using specific stains and immunolabeling techniques. In S. compactum, chlorocysts and hyalocysts exhibit distinct polysaccharide compositions, with methylesterified and demethylesterified homogalacturonans, arabinans, and hemicelluloses contributing to water retention, structural integrity, and photosynthetic efficiency. In contrast, M. polymorpha demonstrates a simpler yet polarized distribution of homogalacturonans, arabinans, mannans, and xyloglucans, with arabinogalactan proteins uniquely localized in rhizoids, improving their flexibility and anchorage to the substrate. Cellulose was uniformly distributed throughout all tissues in both bryophytes, while crystalline cellulose was only faintly observed. These findings highlight how cell wall adaptations contribute to ecological specialization, providing insights into the evolutionary innovations that enable bryophytes to thrive in terrestrial environments. Full article

The ultrastructure of submandibular gland (SMG) of Bactrian camels was observed by a transmission electron microscope. Routine HE staining, special staining combined with immunohistochemistry, and immunofluorescence techniques were used to study the histochemical characteristics of the submandibular gland and the localisation and distribution characteristics of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR). HE results showed that the submandibular gland of Bactrian camels was composed of mixed serous and mucinous acini glands. The submandibular striated duct was highly developed and connected with intercalated ducts with larger diameter. Intercalated ducts are shorter and directly connected to acini. In AB-PAS staining, it was observed that the inner wall of striated tube was strongly positive for AB staining. The distribution of the reticular fibres around the follicles and ducts of the submandibular gland is distinct, with collagen fibres distributed mainly in the periphery of the ducts and sparse collagen fibres in the periphery of the acini. Immunohistochemistry and fluorescence show that EGF is strongly positive in striated and intercalated ducts, and EGFR is weakly positive in striated and intercalated ducts. Bactrian camel SMGs secrete more acidic mucins, and EGF and EGFR were mainly secreted and play a role in the pipeline system of SMGs. Full article

Spheroids, as three-dimensional (3D) cell aggregates, can be prepared using various methods, including hanging drops, microwells, microfluidics, magnetic manipulation, and bioreactors. However, current spheroid manufacturing techniques face challenges such as complex workflows, the need for specialized personnel, and poor batch reproducibility. In this study, we designed a support-free, 3D-printed microwell chip and developed a compatible low-cell-adhesion process. Through simulation and experimental validation, we rapidly optimized microwell size and the coating process. We successfully formed three types of spheroids—human immortalized epidermal cells (HaCaTs), umbilical cord mesenchymal stem cells (UC-MSCs), and human osteosarcoma cells (MG63s)—on the chip. Fluorescent viability staining confirmed the biocompatibility and reliability of the chip. Finally, drug response experiments were conducted using the chip. Compared to traditional methods, our proposed strategy enables high-throughput production of size-controlled spheroids with excellent shape retention, while enhanced gas exchange during culture improves differentiation marker expression. This platform provides an efficient and cost-effective solution for biosensing applications, such as drug screening, disease modeling, and personalized therapy monitoring. Furthermore, the chip shows significant potential for real-time in vitro monitoring of cellular viability, reaction kinetics, and drug sensitivity, offering valuable advancements in biosensor technology for life sciences and medical applications. Full article

Intra-tumor heterogeneity (ITH) is a fundamental characteristic of breast cancer (BC), influencing tumor progression, prognosis, and therapeutic responses. However, the complexity of ITH in BC makes its accurate characterization challenging. This study leverages deep learning (DL) techniques to comprehensively evaluate ITH in early-stage luminal BC and provide a nuanced understanding of its impact on tumor behavior and patient outcomes. A large cohort (n = 2561) of early-stage luminal BC was evaluated using whole slide images (WSIs) of hematoxylin and eosin-stained slides of excision specimens. Morphological features of both the tumor and stromal components were meticulously annotated by a panel of pathologists in a subset of cases. A DL model was applied to develop an algorithm to assess the degree of heterogeneity of various morphological features per individual case utilizing defined patches. The results of extracted features were used to generate an overall heterogeneity score that was correlated with the clinicopathological features and outcome. Overall, 162 features were quantified and a significant positive correlation between these features was identified. Specifically, there was a significant association between a high degree of intra-tumor heterogeneity and larger tumor size, poorly differentiated tumors, highly proliferative tumors, tumors of no special type (NST), and those with low estrogen receptor (ER) expression. When all features are considered in combination, a high overall heterogeneity score was significantly associated with parameters characteristic of aggressive tumor behavior, and it was an independent predictor of poor patient outcome. In conclusion, DL models can be used to accurately decipher the complexity of ITH and provide extra information for outcome prediction. Full article

Objectives: Presently, data on the vascularization of the superficial musculoaponeurotic system of the face (SMAS) are lacking. Thus, the present study aimed to provide new conclusive data about the topography, density, and relationship of the SMAS blood vessels with other components, namely, the fibrous connective tissue and muscles. Methods: The study included a control lot of 42 cases from the archive of the radiology department. In this group, nuclear magnetic resonance angiography (MRA) was performed in order to identify the main sources of vascular supply. In the second group, tissue samples were collected from the midfacial region of 45 patients from the Oro-Maxillo-Facial and Plastic and Reconstructive Surgery clinics of ‘St. Spiridon’ County Clinical Emergency Hospital, Iasi. These patients received surgery for excision of tumoral formations that did not involve SMAS components. These samples underwent micro-CT analysis, hematoxylin and eosin (HE) staining, as well as immunohistochemical (IHC) staining for collagen type III, muscle tissue, and the vascular endothelium. Results: We discovered the particular way in which the SMAS components interrelate with vascularization and the regional differences between them. We have discovered a new vascular network specific to the SMAS, highlighted by both the micro-CT technique and microscopy on slides with special IHC staining. Significant differences were observed in the topographic arrangement, density, and relationships of the microscopic vasculature across midfacial regions. IHC staining provided morphological and functional information about the structure and vascularization of SMAS. Conclusions: The MRA technique could not detect the structural blood vessels of the SMAS and other methods for their in vivo visualization must be sought. The blood vessels of the SMAS mainly follow the topography of the muscle fibers. From the SMAS layer where they are found, the distribution branches reach the stroma of the region and the hypoderm. Our data can contribute to the development of surgical techniques tailored to each individual patient, as well as the enhancement of methods for stimulating cutaneous angiogenesis, improving scarring in this region, and advancing biotissue engineering techniques. Full article

Accurate and efficient classification and quantification of CD34+ cells are essential for the diagnosis and monitoring of leukemia. Current methods, such as flow cytometry, are complex, time-consuming, and require specialized expertise and equipment. This study proposes a novel approach for the label-free identification of CD34+ cells using a deep learning model and lens-free shadow imaging technology (LSIT). LSIT is a portable and user-friendly technique that eliminates the need for cell staining, enhances accessibility to nonexperts, and reduces the risk of sample degradation. The study involved three phases: sample preparation, dataset generation, and data analysis. Bone marrow and peripheral blood samples were collected from leukemia patients, and mononuclear cells were isolated using Ficoll density gradient centrifugation. The samples were then injected into a cell chip and analyzed using a proprietary LSIT-based device (Cellytics). A robust dataset was generated, and a custom AlexNet deep learning model was meticulously trained to distinguish CD34+ from non-CD34+ cells using the dataset. The model achieved a high accuracy in identifying CD34+ cells from 1929 bone marrow cell images, with training and validation accuracies of 97.3% and 96.2%, respectively. The customized AlexNet model outperformed the Vgg16 and ResNet50 models. It also demonstrated a strong correlation with the standard fluorescence-activated cell sorting (FACS) technique for quantifying CD34+ cells across 13 patient samples, yielding a coefficient of determination of 0.81. Bland–Altman analysis confirmed the model’s reliability, with a mean bias of −2.29 and 95% limits of agreement between 18.49 and −23.07. This deep-learning-powered LSIT offers a groundbreaking approach to detecting CD34+ cells without the need for cell staining, facilitating rapid CD34+ cell classification, even by individuals without prior expertise. Full article

Background: One of the crucial factors influencing the longevity of anterior aesthetic dental restorations is the colour stability of resin composites. Bleaching and whitening have become popular methods for enhancing dental aesthetics. Home whitening techniques, such as special pens, are widely available commercially. This in vitro study aimed to determine the efficiency of a whitening pen in removing tea stains from resin composite by measuring colour differences (ΔE₀₀). Additionally, the study aimed to evaluate the variations in colour parameters measured by extra-oral and intra-oral spectrophotometers. Methods: A total of 45 disc-shaped resin composite specimens were randomly divided into three groups; Group 1: stored in artificial saliva (control), Group 2: stored in artificial saliva followed by a whitening pen application, and Group 3: stored in tea followed by a whitening pen application. Colour measurements were taken three times for each specimen using two spectrophotometers (extra-oral and intra-oral devices); T1: before storage (baseline), T2: after storage in artificial saliva or tea for 6 days; and T3: after one week of whitening pen application in groups 2 and 3. The data were statistically analyzed using one-way ANOVA followed by the Tukey post hoc test (p ≤ 0.05). The independent sample t-test was also employed. The equation of CIEDE2000 (ΔE₀₀) was used to calculate the colour difference between the dry, as-prepared specimens (baseline), and those after storage or bleaching. The colour changes exceeding the acceptability threshold (∆E₀₀ = 1.8) were considered unacceptable. Results: After whitening, the colour of the specimens stored in brewed tea (Group 3) remained unacceptable, as indicated by both the extra-oral and intra-oral spectrophotometers (ΔE₀₀ = 4 and 2.9, respectively). Groups 1 and 2 exhibited lower ΔE₀₀ values than Group 3 (p = 0.01 *). No significant difference was observed between Group 1 (stored in artificial saliva) and Group 2 (stored in artificial saliva and then bleached) (p = 0.3). Significant differences were consistently observed between the data obtained from the extra-oral spectrophotometer and the intra-oral one. Conclusions: The whitening pen proved ineffective in removing tea stains from resin composites. Although significant differences were found between the values obtained by the two spectrophotometers (extra-oral and intra-oral), both devices confirmed the unacceptable colour of the tea-stained resin composites after whitening. Full article