Microscopic Characterization of the Mucous Cells and Their Mucin Secretions in the Alimentary Canal of the Blackmouth Catshark Galeus melastomus (Chondrichthyes: Elasmobranchii)

Sharks belong to the most primitive group of jawed vertebrates and have some special structural and functional features such as a cartilaginous skeleton, a spiral intestinal valve, and a rectal gland for osmoregulation. In January 2020, ten specimens of Galeus melastomus, the Blackmouth catshark, were collected from the Gulf of Asinara (North Sardinia, Italy) and the entire alimentary canal was studied using histochemical reactions to characterize the mucous cell types. In the alimentary canal of G. melastomus, mucous cells mainly secrete a mixture of acidic and neutral mucins. Of the acidic mucins, only the carboxylated type was present in mucous cells of the stomach, while the sulfated type predominated in the esophagus and the intestines. The use of lectins revealed a distribution of sugar residues in mucins related to cellular activities of the different regions of the catshark alimentary canal. The current study is the first report to characterize the intestinal mucous cells of G. melastomus and to provide quantitative data on their different populations in the alimentary canal.

Haematological and Histopathological Examinations of African Catfish (Clarias gariepinus) Exposed to Sub-Lethal Concentrations of Paraquat

The study was designed to investigate the sub-lethal effects of paraquat on haematological parameters and histopathology of the gills, skin and liver of Clarias gariepinus juveniles of mean weight (38.26±1.20g) and length (17.50±1.55cm). The fish were exposed to 10, 20, 30, 40 and 50% of the 96hrsLC50 value of 107mg/l estimated from the 96 hours acute toxicity test. Blood samples were collected into heparinized tubes for the analyses of some haematological parameters, while the gills, skin and liver were also removed for histological examinations following standard procedures. The result revealed a significant reduction (P < 0.05) in the values of red blood cells (RBCs), haemoglobin (Hgb), packed cell volume (PCV) and erythrocytes indices from the control. The white blood cells (WBCs) and platelets (Plt) were however increasing significantly (P < 0.05) from those of the control as the test concentrations increased. The histology of the gills revealed some alterations such as epithelial proliferation, vacuolation of the mucus, hyperplasia of epithelial tissue of the gill filament, lifting and necrosis of the secondary lamellae. The exposed skin showed mucous cell proliferation, erosion of the epithelial lining, hypertrophy, necrosis of epithelial cells and widening of the epidermal and dermal layers. The liver exhibited cellular proliferation, sinusoid enlargement, congestion of the central vein, paranchymatous degeneration, vacuolar degeneration, pyknotic nucleic degeneration, legions and necrosis with severity as concentration of paraquat increases. The gills and skin were observed to be the most affected tissues in this study. The study also revealed that paraquat was toxic to C. gariepinus and causes some haematological and histopathological alterations in the fish blood and tissues at concentrations higher than 30mg/l. Therefore, the use of paraquat by farmers should be regulated particularly in area close to the aquatic environment.

Multiomics Provide Insights into the Key Molecules and Pathways Involved in the Physiological Adaptation of Atlantic Salmon (Salmo salar) to Chemotherapeutic-Induced Oxidative Stress

Although chemotherapeutics are used to treat infections in farmed fish, knowledge on how they alter host physiology is limited. Here, we elucidated the physiological consequences of repeated exposure to the potent oxidative chemotherapeutic peracetic acid (PAA) in Atlantic salmon (Salmo salar) smolts. Fish were exposed to the oxidant for 15 (short exposure) or 30 (long exposure) minutes every 15 days over 45 days. Unexposed fish served as the control. Thereafter, the ability of the remaining fish to handle a secondary stressor was investigated. Periodic chemotherapeutic exposure did not affect production performance, though survival was lower in the PAA-treated groups than in the control. Increased ventilation, erratic swimming, and a loss of balance were common behavioural manifestations during the oxidant exposure. The plasma reactive oxygen species levels increased in the PAA-treated groups, particularly after the third exposure, suggesting an alteration in the systemic oxidative stress status. Plasma indicators for internal organ health were affected to a certain degree, with the changes mainly observed after the second and third exposures. Metabolomics disclosed that the oxidant altered several circulating metabolites. Inosine and guanosine were the two metabolites significantly affected by the oxidative stressor, regardless of exposure time. A microarray analysis revealed that the gills and liver were more responsive to the oxidant than the skin, with the gills being the most sensitive. Moreover, the magnitude of the transcriptomic modifications depended on the exposure duration. A functional analysis showed that genes involved in immunity and ribosomal functions were significantly affected in the gills. In contrast, genes crucial for the oxidation-reduction process were mainly targeted in the liver. Skin mucus proteomics uncovered that the changes in the mucosal proteome were dependent on exposure duration and that the oxidant interfered with ribosome-related processes. Mucosal mapping revealed gill mucous cell hypertrophy after the second and third exposures, although the skin morphological parameters remained unaltered. Lastly, repeated oxidant exposures did not impede the ability of the fish to mount a response to a secondary stressor. This study provides insights into how a chemical oxidative stressor alters salmon physiology at both the systemic and mucosal levels. This knowledge will be pivotal in developing an evidence-driven approach to the use of oxidative therapeutics in fish, with some of the molecules and pathways identified as potential biomarkers and targets for assessing the physiological cost of these treatments.

Corticosteroid Insensitivity Persists in the Absence of STAT1 Signaling in Severe Allergic Airway Inflammation

Corticosteroid insensitivity in asthma limits the ability to effectively manage severe asthma, which is characterized by persistent airway inflammation, airway hyperresponsiveness (AHR), and airflow obstruction despite corticosteroid treatment. Recent reports indicate that corticosteroid insensitivity is associated with increased interferon-gamma (IFN-g) levels and T-helper (Th) 1 lymphocyte infiltration in severe asthma. Signal Transducer and Activator of Transcription 1 (STAT1) activation by IFN-g is a key signaling pathway in Th1 inflammation, however its role in the context of severe allergic airway inflammation and corticosteroid sensitivity remains unclear. In the present study, we challenged wild type (WT) and Stat1-/- mice with mixed allergens (MA) augmented with c-di-GMP, an inducer of Th1 cell infiltration with increased eosinophils, neutrophils, Th1, Th2, and Th17 cells. Compared to WT mice, Stat1-/- had reduced neutrophils, Th1 and Th17 cell infiltration. To evaluate corticosteroid sensitivity, mice were treated with either vehicle, 1 or 3 mg/kg fluticasone propionate (FP). Corticosteroid significantly reduced eosinophil infiltration and cytokine levels in both c-di-GMP + MA-challenged WT and Stat1-/- mice. However, histological and functional analyses show that corticosteroids did not reduce airway inflammation, epithelial mucous cell abundance, airway smooth muscle mass, and AHR in c-di-GMP + MA-challenged WT or Stat1-/- mice. Collectively, our data suggest that increased Th1 inflammation is associated with a decrease in corticosteroid sensitivity. However, increased airway pathology and AHR persist in the absence of STAT1 indicate corticosteroid insensitivity in structural airway cells is a STAT1 independent process.

Irg1/itaconate metabolic pathway is a crucial determinant of dendritic cells immune-priming function and contributes to resolute allergen-induced airway inflammation

Itaconate is produced from the mitochondrial TCA cycle enzyme aconitase decarboxylase (encoded by immune responsive gene1; Irg1) that exerts immunomodulatory function in myeloid cells. However, the role of the Irg1/itaconate pathway in dendritic cells (DC)-mediated airway inflammation and adaptive immunity to inhaled allergens, which are the primary antigen-presenting cells in allergic asthma, remains largely unknown. House dust mite (HDM)-challenged Irg1−/− mice displayed increases in eosinophilic airway inflammation, mucous cell metaplasia, and Th2 cytokine production with a mechanism involving impaired mite antigen presentations by DC. Adoptive transfer of HDM-pulsed DC from Irg1-deficient mice into naïve WT mice induced a similar phenotype of elevated type 2 airway inflammation and allergic sensitization. Untargeted metabolite analysis of HDM-pulsed DC revealed itaconate as one of the most abundant polar metabolites that potentially suppress mitochondrial oxidative damage. Furthermore, the immunomodulatory effect of itaconate was translated in vivo, where intranasal administration of 4-octyl itaconate 4-OI following antigen priming attenuated the manifestations of HDM-induced airway disease and Th2 immune response. Taken together, these data demonstrated for the first time a direct regulatory role of the Irg1/itaconate pathway in DC for the development of type 2 airway inflammation and suggest a possible therapeutic target in modulating allergic asthma.

Gill Histopathology as a Biomarker for Discriminating Seasonal Variations in Water Quality

Histopathological alterations in various fish organs have a pronounced value in aquatic toxicology and are widely used in environmental monitoring. The aim of this study was to evaluate whether histopathological alterations in fish gills can discriminate seasonal variations in environmental conditions within the same aquatic ecosystem, and if so, which alterations contributed the most to seasonal differentiation. Microscopic examination of common bream Abramis brama gills displayed various alterations in gill structure, including epithelial hypertrophy, hyperplasia, mucous and chloride cell alterations, epithelial lifting, necrosis, hyperemia and aneurism. These alterations were subsequently quantified by a semi-quantitative analysis in order to detect differences in the intensity of the mentioned alterations. Epithelial hypertrophy, hyperplasia, epithelial lifting and necrosis varied significantly between seasons with only necrosis being significantly higher in the first season. Discriminant canonical analysis displayed that epithelial hyperplasia, mucous cell alterations, epithelial lifting and necrosis contributed the most to discrimination between seasons. Overall, this study demonstrates that histopathological biomarkers in fish gills can be used in discriminating seasonal variations in water quality within the same aquatic ecosystem.

Respiratory Nasal Mucosa in Chronic Rhinosinusitis with Nasal Polyps versus COVID-19: Histopathology, Electron Microscopy Analysis and Assessing of Tissue Interleukin-33

(1) Background: Chronic rhinosinusitis with nasal polyps (CRSwNP) is one of the most studied rhinological disorders. Modifications of the respiratory nasal mucosa in COVID-19 patients are so far unknown. This paper presents a comparative morphological characterization of the respiratory nasal mucosa in CRSwNP versus COVID-19 and tissue interleukin (IL)-33 concentration. (2) Methods: We analyzed CRSwNP and COVID-19 samples through histopathology, scanning and transmission electron microscopy and performed proteomic determination of IL-33. (3) Results: Histopathologically, stromal edema (p < 0.0001) and basal membrane thickening (p = 0.0768) were found more frequently in CRSwNP than in COVID-19. Inflammatory infiltrate was mainly eosinophil-dominant in CRSwNP and lymphocyte-dominant in COVID-19 (p = 0.3666). A viral cytopathic effect was identified in COVID-19. Scanning electron microscopy detected biofilms only in CRSwNP, while most COVID-19 samples showed microbial aggregates (p = 0.0148) and immune cells (p = 0.1452). Transmission electron microscopy of CRSwNP samples identified biofilms, mucous cell hyperplasia (p = 0.0011), eosinophils, fibrocytes, mastocytes, and collagen fibers. Extracellular suggestive structures for SARS-CoV-2 and multiple Golgi apparatus in epithelial cells were detected in COVID-19 samples. The tissue IL-33 concentration in CRSwNP (210.0 pg/7 μg total protein) was higher than in COVID-19 (52.77 pg/7 μg total protein) (p < 0.0001), also suggesting a different inflammatory pattern. (4) Conclusions: The inflammatory pattern is different in each of these disorders. Results suggested the presence of nasal dysbiosis in both conditions, which could be a determining factor in CRSwNP and a secondary factor in COVID-19.

Smoking shifts human small airway epithelium club cells toward a lesser differentiated population

The club cell, a small airway epithelial (SAE) cell, plays a central role in human lung host defense. We hypothesized that subpopulations of club cells with distinct functions may exist. The SAE of healthy nonsmokers and healthy cigarette smokers were evaluated by single-cell RNA sequencing, and unsupervised clustering revealed subpopulations of SCGCB1A1+KRT5loMUC5AC− club cells. Club cell heterogeneity was supported by evaluations of SAE tissue sections, brushed SAE cells, and in vitro air–liquid interface cultures. Three subpopulations included: (1) progenitor; (2) proliferating; and (3) effector club cells. The progenitor club cell population expressed high levels of mitochondrial, ribosomal proteins, and KRT5 relative to other club cell populations and included a differentiation branch point leading to mucous cell production. The small proliferating population expressed high levels of cyclins and proliferation markers. The effector club cell cluster expressed genes related to host defense, xenobiotic metabolism, and barrier functions associated with club cell function. Comparison of smokers vs. nonsmokers demonstrated that smoking limited the extent of differentiation of all three subclusters and altered SAM pointed domain-containing Ets transcription factor (SPDEF)-regulated transcription in the effector cell population leading to a change in the location of the branch point for mucous cell production, a potential explanation for the concomitant reduction in effector club cells and increase in mucous cells in smokers. These observations provide insights into both the makeup of human SAE club cell subpopulations and the smoking-induced changes in club cell biology.

Gross and Histopathological Lesion Induced on the Gills Ofbarbus Fish by Monogenea Trematods Habiteting in Western Part of Lake Tana, Amhara Region, Ethiopia

Back ground: Gill parasitic is common on cultured and wild fish. Many of these species have long been recognized to have the potential to affect the growth, fecundity and survival of hosts.the objective of the study were to determine the prevalence and identify, gross and histopathological lesion of gills caused by Dactylogyrus and Gryodactylus spp. parasitic infection on naturally infected barbus fish.Result: A cross sectional study was conducted from in Lake Tana, amhara region. Three hundred eight four gill specimens were collected from barbus fish and of these, 22 (5.7%) of fish were infected with monogenean parasites. Higher prevalence of Dactylogyrus spp. (4.86%) was recorded than Gryodactylus sp (1.04). Large size fishes (≥ 20cm) were more susceptible to Dactylogyrus spp and Gryodactylus spp. However, highest prevalence of Dactylogyrus sp. was found in large fish size (6.85%). Gryodactylus sp. was not recorded in all small size fish. Descriptive statistics such as percentages was used to describe the nature and the characteristics of the disease. At gross examination of gills, hyperaemia and swollen, excess mucus secretion, paleness, congestion of branchial blood vessel were observed. Gills specimens of infected fish were fixed with10% formalin, for further identification and stained with the haematoxylin-eosin. Histopathological changes included hyperplasia, congestion and mucous cell proliferation of the gill epithelium and damaged primary and secondary lamellae, the uplifting of respiratory epithelial wall and damaged pillar cells. Conclusion: Gross and histopathological changes induced by the Dactylogyrus spp and Gryodactylus spp of parasites adversely affected the proper functioning of the gills of the host fish which can lead to detrimental effect on the health status of the fish and may result in huge economic losses through increased mortality.

Vaginal mucus in mice: Developmental and gene expression features of epithelial mucous cells during pregnancy

The vagina is the site of copulation and serves as the birth canal. It also provides protection against external pathogens. In mice, due to the absence of cervical glands, the vaginal epithelium is the main producer of vaginal mucus. The development and differentiation of vaginal epithelium-constituting cells and the molecular characteristics of vaginal mucus have not been thoroughly examined. Here, we characterized vaginal mucous cell development and the expression of mucus-related factors in pregnant mice. The vaginal mucous epithelium layer thickened and became multilayered after day 12 of pregnancy and secreted increasing amounts of mucus until early postpartum. Using histochemistry and transmission electron microscopy, we found supra-basal mucous cells as probable candidates for precursor cells. In vaginal mucous cells, the expression of TFF1, a stabilizer of mucus, was high, and some members of mucins and antimicrobial peptides (MUC5B and DEFB1) were expressed in a stage-dependent manner. In summary, this study presents the partial characterization of vaginal epithelial mucous cell lineage and expression of genes encoding several peptide substances that may affect vaginal tissue homeostasis and mucosal immunity during pregnancy and parturition.