Search Results (450)

New Zealand abalone (Haliotis iris) holds ecological, economic, and cultural value, with wild stocks supporting fisheries and an emerging aquaculture industry. Wild-caught adult abalone are often used as broodstock, but captivity can affect spawning and offspring quality. This study is the first to profile wild and farmed H. iris broodstock using histology, proximate composition, microbiome, and metabolomics analyses. Histology showed higher gonadal abnormalities in farmed abalone, while wild abalone exhibited increased ciliates in their gills, indicating richer marine–microorganism interactions. Microbiome analyses revealed a higher microbial richness and diversity in the buccal cavity of wild abalone. The core microbiota phyla across both groups included Proteobacteria, Bacteroidota, Campylobacterota, Fusobacteria, and Firmicutes. Proximate analyses showed higher muscle protein in farmed abalone, while gonadal tissue partitioned by sex showed higher fat in females and higher protein in males. Metabolomics revealed altered amino acid metabolism in the adductor muscle, carboxylic acid metabolism in the gonad, and fatty acid metabolism in the foot. This investigation expands our understanding of the physiological and microbial differences between wild and farmed abalone, showing altered gonad and muscle conditions from prolonged captivity and highlighting the need for greater microbial diversity in cultured stocks. Full article

Ciliate infestations in aquatic organisms are commonly associated with aquaculture, yet their impact on natural ecosystems remains largely understudied. This study describes a mobilid peritrich species infesting the gills of Octopus bimaculatus from the Gulf of California, Mexico. All 76 examined hosts (100%) exhibited infestation, with a mean intensity of 687 ± 228 (279–1077) urceolariid cells per gill. The ciliate cells displayed morphological traits consistent with those of the genus Urceolaria: turban-shaped cells measuring 44.2 ± 13.2 (31.3–88.6) µm in diameter; an adhesive disc of 36.5 ± 10.7 (29.2–74.6) μm in diameter; 18–19 plates measuring 11.0 ± 0.86 (9–12) µm in length; and 166–169 radial pins. Phylogenetic analysis of 18S rDNA sequences placed this species within the genus Urceolaria, a sister group to Urceolaria urechi and Urceolaria serpularum, with a genetic distance of 1.0% with respect to the previously described species. Combined morphological and molecular data support the description of a new species, Urceolaria carmenae n. sp. This is the first record of a mobilid peritrich in cephalopod mollusks, thereby enhancing our understanding of the diversity of ciliates among marine invertebrates in their natural habitats. Full article

We hereby present a case of a 22-year-old female patient with a lesion on the right side of the uvula. Histopathological assessment identified a cystic lesion with a branching configuration. The cyst lining consisted of non-keratinising stratified squamous epithelium, ciliated columnar epithelium, and focal areas of mucinous columnar epithelium. Within the cystic lumen, a focus of ectopic parathyroid tissue was observed. Based on these findings, a final diagnosis of a uvular branchial cleft cyst containing ectopic parathyroid tissue was established. Pathological lesions of the uvula are rare, and cystic lesions typically correspond to epidermoid cysts. This case, therefore, represents the first documented occurrence of a uvular branchial cleft cyst containing ectopic parathyroid tissue, underscoring the importance of detailed histopathological and immunohistochemical evaluation in the assessment of even small, incidentally detected uvular lesions. Full article

Background/Objectives: β-Carotene oxygenase-2 (BCO2) is a mitochondrial carotenoid-cleaving enzyme expressed in multiple tissues, including the lungs. While BCO2 regulates carotenoid handling, its role in shaping pulmonary immune architecture and antiviral responses is unknown. We hypothesized that BCO2 deficiency reprograms epithelial–innate circuits and alters antiviral outcomes. Methods: BCO2-knockout (KO) and C57BL/6J wild-type (WT) mice underwent lung single-cell RNA sequencing (scRNA-seq), immunoblotting, and intranasal SARS-CoV-2 challenge to assess cell-type heterogeneity, pathway programs (by gene set variation analysis, GSVA), and antiviral responses. Results: scRNA-seq resolved 14 major lung cell populations with cell-type-specific pathway shifts. Compared with WT, BCO2 KO lungs showed increased conventional dendritic cells and natural killer (NK) cells, with reductions in macrophages, B cells, and endothelial cells. In KO alveolar type II cells, GSVA indicated a stress-adapted metabolic program. Ciliated epithelium exhibited vitamin-K-responsive and axoneme-remodeling signatures with attenuated glucocorticoid and very-low-density lipoprotein remodeling. Innate lymphoid type 2 cells favored fatty acid oxidation and chromatin dynamics with reduced mitochondrial activity. NK cells were biased toward constitutive chemokine/cytokine secretion and counter-inflammatory signaling. Immunoblotting confirmed the elevated level of interferon regulatory factor-3 protein in BCO2-KO lungs. Functionally, BCO2-KO mice had improved outcomes after intranasal SARS-CoV-2 exposure. Conclusions: Loss of BCO2 reconfigures the pulmonary immune landscape and enhances antiviral responsiveness in mice. These findings identify BCO2 as a nutrient-linked enzyme with immunomodulatory impact and highlight cell-state changes as candidate mechanisms for improved antiviral tolerance. Full article

Antarctic microorganisms have developed extraordinary strategies for adaptation. They have also demonstrated the ability to produce various biopolymers in response to environmental stress. The demand for biopolymers is constantly increasing and is expected to grow further. Among emerging biomaterials, bacterial cellulose (BC) is generating significant interest due to its unique characteristics that distinguish it from plant-based cellulose. BC exhibits higher purity, water-holding capacity, and tensile strength compared to its plant-based counterpart. Furthermore, BC can be obtained through environmentally friendly protocols. Several bacterial strains have already been identified as cellulose producers, including Komagataeibacter xylinus. In this study, a marine bacterial strain named Pseudomonas sp. ef1, isolated from a consortium associated with the Antarctic ciliate Euplotes focardii, was tested for cellulose production. We found that this Antarctic Pseudomonas can produce BC in conditions that appear unique to this bacterial strain. Furthermore, the final BC product is structurally different from that obtained from the well-known BC producer Komagataeibacter xylinus. Additionally, a putative cellulose synthase was identified from the Pseudomonas sp. ef1 genome, exhibiting unique characteristics that may account for the unique BC production capability of this Antarctic marine Pseudomonas. The versatility of BC opens numerous applications, including in papermaking, food, pharmaceutical, and biomedical sectors. Full article

In this study, we evaluated the cytotoxicity and antioxidant activity of the soil ciliate Rigidohymena tetracirrata (Gellért, 1942) Berger 2011, exposed to single and bimetallic mixtures of heavy metals (HMs) for 24 h. Ecotoxicological tests showed LC₂₀ values of 0.16, 19.86 and 0.68 mg L⁻¹ to Copper (Cu), Zinc (Zn), and Cadmium (Cd), respectively, and LC₅₀ values of 0.25, 44.12 and 1.12 mg L⁻¹, respectively. Furthermore, it was observed that the mixture of Cd and Zn exhibited antagonism in comparison to other mixtures, (Cd + Cu and Cu + Zn). In the total phenolic content (TPC) assay, a higher phenolic content was observed for the LC₂₀ of extracellular Cu (p ≤ 0.01) and the LC₂₀ of intracellular Cd (p ≤ 0.001). The LC₅₀ values for Cd and Zn in both extracellular and intracellular contents demonstrated increased α,α-diphenyl-β-picrylhydrazyl (DPPH) scavenging activity with significant values of p ≤ 0.05, respectively. Regarding hydroxyl scavenging activity (HRSA), the LC₅₀ of extracellular Cd (p ≤ 0.001) and LC₅₀ of intracellular Cu (p ≤ 0.001) exhibited higher antioxidant activity. Therefore, the present study suggests that R. tetracirrata holds considerable potential as bioindicators and could be used as a model organism in ecotoxicological studies of soil polluted by HMs. Full article

Air–liquid interface (ALI) cultures offer a physiologically relevant in vitro model of the airway epithelium (AE), capable of recapitulating key structural and functional features observed in vivo. In this study, we established and validated a murine ALI culture system comprising pseudostratified epithelia with functional tight junctions, ciliated cells and goblet cells. To assess their innate immune functions, we designed and 3D-printed an autoclavable aerosol deposition chamber, which allowed us to expose differentiated AE cultures to house dust mite (HDM) allergen. Upon HDM exposure, AE cells mounted a time-dependent innate immune response characterized by the secretion of complement component C3, the generation of its active cleavage products C3a and increased expression of C3aR and C5aR1. This was associated with increased intracellular TSLP and IL-25 production and TSLP release in AE cells. Progressive loss of tight junction integrity and reduced transepithelial electrical resistance (TEER) demonstrated epithelial susceptibility to allergen protease-induced cell damage. Together, we established a murine ALI system preserving airway epithelial architecture and a nebulization system to study innate immune activation of AE cells in response to HDM mimicking the initial phase of allergen sensitization. More generally, we described a powerful and accessible platform for studying epithelial-driven mechanisms in murine airway immune responses. Full article

This study aimed to investigate the histological features of the intestine of Acipenser dabryanus from 1 to 15 months of age via HE staining, AB-PAS staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The intestine of A. dabryanus comprises the duodenum, spiral valve intestine, and rectum. With age, the duodenal diameter and mucosal/muscular layer thickness increased, the spiral valve intestine’s mucosa thickened and protrusions formed networks, and the rectal diameter enlarged. Abundant mucus cells, predominantly type IV, were found in the duodenum, spiral valve intestine, and rectum of A. dabryanus at different ages by AB-PAS staining. Our study confirmed the presence of ciliated columnar cells (with ‘9 + 2’ cilia structure) with orderly arranged cilia at their apices in the mucosal epithelium of A. dabryanus’s duodenum, spiral valve intestine, and rectum for the first time, as shown by SEM and TEM. The presence of spiral valves and ciliated columnar cells in the intestinal structure of A. dabryanus highlights its unique features and evolutionary significance. These findings highlight A. dabryanus’s unique intestinal features and evolutionary significance, providing a basis for scientific feed formulation and enhancing our understanding of the histological characteristics of the sturgeon intestine. Full article

A straightforward ex vivo approach has been developed and refined to enable high-resolution imaging and quantitative assessment of motile cilia function in mouse airway epithelial tissue, allowing critical insights into cilia motility and cilia generated flow using different mouse models or following different sample treatments. In this method, freshly excised mouse trachea is cut longitudinally through the trachealis muscle which is then sandwiched between glass coverslips within a thin silicon gasket. By orienting the tissue along its longitudinal axis, the natural curling of the trachealis muscle helps maintain the sample in a configuration optimal for imaging along the full tracheal length. High-speed video microscopy, utilizing differential interference contrast (DIC) optics and a fast digital camera capturing at >200 frames per second is then used to record ciliary motion. This enables detailed measurement of both cilia beat frequency (CBF) and waveform characteristics. The application of 1 µm microspheres to the bathing media during imaging allows for additional analysis of fluid flow generated by ciliary activity. The entire procedure typically takes around 40 min to complete per animal: ~30 min for tissue harvest and sample mounting, then ~10 min for imaging samples and acquiring data. Full article

This study investigates the defensive mechanisms employed by the freshwater flatworm Stenostomum sphagnetorum against the predatory ciliate, Coleps hirtus. Focusing on the role of the glandular secretions produced by S. sphagnetorum, the research demonstrates that the flatworm secretes mucus that acts as a barrier, disrupting predator attack. In particular, we show that: (1) S. sphagnetorum specimens artificially deprived of glandular secretions are significantly more susceptible to predation by C. hirtus than untreated specimens; (2) the secretion-deprived organisms consistently exhibit a significantly greater sensitivity to the main toxins employed by C. hirtus for predation, relative to untreated counterparts; (3) the characterization of the glandular secretion indicates that the mucus contains both neutral and acidic glycosaminoglycans, along with protein components, suggesting a complex chemical composition that may contribute to its protective function. Full article

Cholesterol is an essential plasma membrane component, and altered cholesterol metabolism has been linked to cholesterol accumulation in the airways of COPD and cystic fibrosis patients. However, its role in airway epithelial differentiation is not well understood. Tandem mass spectrometry-based proteomic analysis of differentiating primary human bronchial epithelial cells (phBECs) revealed an overall inhibition of the cholesterol biosynthesis pathway. We hypothesized that excess cholesterol impairs the differentiation of phBECs into a fully functional bronchial epithelium. PhBECs were differentiated in the presence of 80 µM cholesterol for 21 days, the main airway cell type populations monitored using qRT-PCR and immunofluorescent stainings, and epithelial barrier integrity was analyzed via transepithelial electrical resistance measurements. Chronic cholesterol exposure led to a significant increase in CC10⁺ secretory cells at the expense of ciliated cells. Pathway enrichment analysis suggested the tumor protein p53 as a master regulator of genes during normal differentiation of phBECs. Chronic cholesterol exposure drastically impaired the nuclear translocation of p53. Our findings suggest that this inhibition underlies the cholesterol-induced expansion of CC10⁺ secretory cell populations at the expense of ciliated cells. In conclusion, we identify cholesterol as an important regulator of normal bronchial epithelial cell differentiation through inhibition of p53 nuclear translocation. Full article

Objective: This study aims to identify clinically relevant lactylation-related biomarkers in chronic obstructive pulmonary disease (COPD) and investigate their potential mechanistic roles in COPD pathogenesis. Methods: Differentially expressed genes (DEGs) were identified from the GSE21359 dataset, followed by weighted gene co-expression network analysis (WGCNA) to detect COPD-associated modules. Least absolute shrinkage and selection operator (LASSO) regression and support vector machine–recursive feature elimination (SVM–RFE) algorithms were applied to screen lactylation-related biomarkers, with diagnostic performance evaluated through the ROC curve. Candidates were validated in the GSE76925 dataset for expression and diagnostic robustness. Immune cell infiltration patterns were exhibited using EPIC deconvolution. Single-cell transcriptomics (from GSE173896) were processed via the ‘Seurat’ package encompassing quality control, dimensionality reduction, and cell type annotation. Cell-type-specific markers and intercellular communication networks were delineated using the ‘FindAllMarkers’ package and the ‘CellChat’ R package, respectively. In vitro validation was conducted using a cigarette smoke extract (CSE)-induced COPD model. Results: Integrated transcriptomic approaches and multi-algorithm screening (LASSO/Boruta/SVM–RFE) revealed carbonyl reductase 1 (CBR1) and peroxiredoxin 1 (PRDX1) as core COPD biomarkers enriched in oxidation–reduction and inflammatory pathways, with high diagnostic accuracy (AUC > 0.85). Immune profiling and scRNA-seq delineated macrophage and cancer-associated fibroblasts (CAFs) infiltration with oxidative-redox transcriptional dominance in COPD. CBR1 was significantly upregulated in T cells, neutrophils, and mast cells; and PRDX1 showed significant upregulation in endothelial, macrophage, and ciliated cells. Experimental validation in CSE-induced models confirmed significant upregulation of both biomarkers via transcription PCR (qRT-PCR) and immunofluorescence. Conclusions: CBR1 and PRDX1 are lactylation-associated diagnostic markers, with lactylation-driven redox imbalance implicated in COPD progression. Full article

Immunofluorescence (IF) microscopy of ciliated epithelium is gaining increased popularity as a pre-genetic diagnostic method in primary ciliary dyskinesia (PCD). Ensuring reliable IF-based diagnostics in PCD requires robust standardization of staining methods and antibody performance. We applied whole slide scanning and automated image analysis to systematically evaluate the influence of various sample storage conditions on the specificity of IF staining. We tested eight polyclonal antibodies targeting diverse axonemal protein epitopes, routinely used for PCD diagnostics, under seven different temperature and time combinations. The storage conditions simulated handling of epithelial brushing on glass slides: after material collection at the clinic, during transport, or after reception at the diagnostic laboratory. Our study revealed that proper slide storage conditions are essential for the reliable PCD diagnosis via IF staining. We suggest continuous storage at −80 °C or −20 °C for slides prepared at the diagnostic laboratory, and storage at −20 °C or 4 °C for slides prepared remotely and shipped. Moreover, the IF sensitivity to slide storage conditions differs among antibodies targeting various ciliary elements, with molecular ruler proteins being particularly sensitive to prolonged storage at room temperature. We emphasize the inclusion of additional control slides to mitigate the inter-individual differences and the crucial correlation of IF results with comprehensive patient clinical history for enhanced diagnostic reliability. Full article

The paper presents the results of a microfossil study of Holocene sediments in the Yana River flow zone in the southeastern part of the Laptev Sea. A rich diatom flora (242 species and intraspecific taxa, of which 177 species are freshwater) was revealed; additionally, five species of marine tintinnids (planktonic ciliates) and 15 species of freshwater testate amoebae (testacean) were discovered for the first time in the sea sediments. Three assemblages of microfossils reflecting the phases of environmental changes during the Holocene transgression are distinguished in the studied sediments of core LV83-32. Assemblage 1 was formed under terrestrial conditions (assemblage of diatoms Eunotia-Pinnularia and testacean Difflugia-Cylindrifflugia-Centropyxis), assemblage 2 in the zone of mixing of sea and fresh waters (assemblages of diatoms Cyclotella striata-Aulacoseira, Thalassiosira hyperborea-Chaetoceros and T. hyperborea-Aulacoseira, testacean Cyclopyxis kahli, tintinnids Tintinnopsis fimbriata), and assemblage 3 reflects modern conditions in the inner shelf of the Laptev Sea under the strong influence of river runoff (assemblage of diatoms T. hyperborea-Aulacoseira-M. arctica and tintinnids Tintinnopsis ventricosoides). Changes in the natural environment in the coastal part of the Laptev Sea shelf during the Holocene, established by microfossil assemblages, are confirmed by geochemical data. Full article

Balantioides coli is the only ciliate currently described as an intestinal parasite of humans, although it can also infect other animals, particularly pigs. Its in vitro cultivation remains challenging, and no axenic culture system is currently available. Cultures are initiated by adding small amounts of feces containing cysts or trophozoites to the culture medium. Implantation success is lower when starting from cysts, and the mechanisms and early events of excystation remain poorly understood. In this study, we describe the sequence of events involved in excystation and identify factors potentially important for culture establishment. Cysts were obtained from orangutan feces and genetically confirmed as B. coli. Only viable cysts, determined by trypan blue or methylene blue exclusion, were used. After artificial digestion with pepsin and trypsin, cysts were incubated at 28 °C for up to 72 h in DMEM supplemented with L-glutamine, yeast extract, fetal bovine serum, and starch granules. Excystation began with a fissure in the cyst wall, allowing for bacterial entry. This appeared to stimulate the trophozoites, the increased motility of which progressively weakened and ruptured the wall, allowing for their emergence. Wall rupture and bacterial entry were critical for activation., whereas starch type had no apparent influence. Excystation occurred within the first hours; otherwise, cysts degenerated. Full article