Superficial Epithelial Cells Research Articles

Effect of endonasal dacryocystorhinostomy on conjunctival morphology and visual quality

ABSTRACT Clinical relevance Endonasal dacryocystorhinostomy (EN-DCR) is one of the preferred interventions for occlusion of the nasolacrimal drainage system. Understanding the bulbar conjunctival changes that occur after EN-DCR surgery will provide better insight into the effect of this procedure on visual quality and patient comfort. Background This study aimed to evaluate visual quality and bulbar conjunctival cytologic changes using in vivo confocal microscopy and corneal topography in patients with epiphora resulting from nasolacrimal duct obstruction after EN-DCR. Methods Nineteen eyes of 19 patients with epiphora were enrolled. All patients underwent EN-DCR with silicone tube intubation. Before and six months after EN-DCR, higher-order aberrations (HOAs) were measured using the Sirius corneal topographer/aberrometer, and bulbar conjunctival changes were assessed using in vivo confocal microscopy. Root mean square values of coma, trefoil, spherical, and total HOAs were analysed. Conjunctival superficial and basal epithelial cell and goblet cell densities and bulbar conjunctival changes were evaluated. The severity of epiphora was assessed by the Munk score before and six months after EN-DCR. Results Nineteen eyes of 19 patients (16 women and 3 men) were examined. The mean age of the patients was 40.8 ± 9.8 years. Total HOA and coma aberrations were significantly decreased at six months after EN-DCR compared to preoperatively. Conjunctival superficial and basal epithelial cell densities and epithelial thickness were significantly increased while goblet cell density was unchanged at six months after EN-DCR. There was also a significant improvement in Munk score six months postoperatively. Conclusion EN-DCR can lead to conjunctival remodelling. Alterations in HOAs and point spread function may occur during the postoperative period. Moreover, point spread function may be positively associated with Munk score.

Single-cell transcriptomics predicts colony stimulating factor-1 regulation of age-dependent intestinal epithelial restitution

Intestinal diseases associated with ischemic injury, which damages the principal barrier against noxious luminal contents, result in unacceptably poor outcomes in newborns. Ischemia-induced loss of the intestinal epithelial barrier predisposes patients to life-threatening sepsis unless that barrier is rapidly restored. There is an age-dependency of intestinal recovery in that neonates are the most susceptible to succumb to disease of the intestinal barrier versus older patients. While this age-dependence in repair has not been demonstrated in traditional rodent models, we have developed a highly translational pig model of intestinal ischemic injury and repair that does reflect this difference. We have shown that, while juvenile (weaned) pigs recover rapidly after ischemic intestinal injury, barrier repair is markedly underdeveloped in neonatal (nursing) pigs due to complete failure of epithelial restitution. Importantly, we found that restitution in neonates can be rescued by the direct application of homogenized mucosa from ischemia-injured small intestine from juvenile pigs. The mechanisms that allow for this restitution and rescue remain to be defined. We hypothesized that by identifying a subpopulation of restituting enterocytes by their expression of cell migration transcriptional pathways, we can then predict novel upstream regulators of age-dependent restitution response programs. Superficial mucosal epithelial cells collected from recovering ischemic jejunum of juvenile pigs were processed for single cell RNA sequencing, unbiased clustering and upstream regulator analysis. A porcine intestinal epithelial cell line (IPEC-J2) and banked tissues from prior rescue experiments were qualitatively and functionally assessed for activity of predicted upstream regulators. Single cell transcriptomics in recovering juvenile epithelium revealed a subcluster of absorptive enterocytes that express several cell migration pathways key to restitution. Differentially expressed genes in this subcluster predicted their upstream regulation by many potential molecules, including colony stimulating factor-1 (CSF-1) which is known to induce cell migration in non-intestinal epithelial tissues. To begin validating this prediction, we demonstrated that CSF-1 was enriched in the ischemic juvenile mucosa which rescues neonatal restitution and documented expression of the CSF-1 receptor (CSF1R) in both neonatal and juvenile epithelium, indicating that these cells are equipped to respond to CSF-1. CSF-1 and CSF1R co-localized in ischemic juvenile, but not neonatal, wound-adjacent epithelial cells and in the restituted epithelium of juveniles and rescued (but not control) neonates. Further, the CSF1R inhibitor BLZ945 reduced restitution in scratch wounded IPEC-J2 cells. Single cell transcriptomics have the power to inform potential novel therapeutic targets, such as CSF-1, to improve mucosal recovery in neonates with intestinal failure in this unique and powerful pig model. NIH K01 OD028207, NIH R01 HD095876, NIH U01 TR002953, NIH T32 OD011130, NIH P30 DK034987, USDA NIFA 1007263 and 07985. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Nucleus-Mitochondria Contact Sites Are Associated With Asynthetic Fission in Zebrafish Skin.

Rapid increase in body surface area of growing zebrafish larvae (Danio rario) is partially accomplished by asynthetic fission of superficial epithelial cells (SECs) of the skin. There are two cycles of this atypical form of cell division which is unaccompanied by DNA replication; resulting in cells with a variable DNA content. Here, electron microscopy of basal epithelium cells that give rise to these SECs in zebrafish larvae shows aggregation of mitochondria around the nucleus and the formation of nucleus-mitochondria membrane contact sites. Membrane aggregates appear in the lumen of the nuclear envelope at these sites of membrane contact in some cells, suggesting lipid turnover in this vicinity. As the epithelial cells mature and stratify, the mitochondria are engulfed by extensions arising from the nuclear envelope. The mitochondrial outer membrane fragments and mitochondria fuse with the nuclear envelope and parts of the endoplasmic reticulum. Other organelles, including the Golgi apparatus, progressively localize to a central region of the cell and lose their integrity. Thus, asynthetic fission is accompanied by an atypical pattern of organelle destruction and a prelude to this is the formation of nucleus-mitochondria membrane contact sites.

An enhanced model for environmental dry eye: Exploring pathological features and underlying factors

This study aimed to develop an enhanced environmental dry eye (EDE) model that accurately reproduces the etiology of prolonged visual fatigue and investigates the underlying pathological features. A total of 40 adult SPF-grade Wistar rats were randomly assigned to control (n = 20) and model (n = 20) groups. Rats in the control group were maintained under normal conditions, while rats in the model group were exposed to a controlled frontal airflow of 2–4 m/s from a fan for 7.5 h daily while placed on a suspended cylindrical wire mesh frame. Various assessments were performed at different time points during the 14-day experiment, including blink frequency, tear secretion (phenol red thread test), tear film breakup time (BUT), fluorescein staining (FL), corneal epithelial status (light microscopy), ultrastructure of corneal epithelial cells (electron microscopy), and expression levels of inflammatory cytokines (IL-1β, TNF-α) in tears (enzyme-linked immunosorbent assay). Additionally, mRNA and protein expression levels of MMP-9, IL1β, IL6, TNF-α, IFN-γ, and caspase-3 in corneal tissues were quantified (real-time quantitative PCR and Western blotting). Compared to the control group, the model group rats exhibited significant decreases in blink frequency (P < 0.001), tear secretion (Schirmer I test) values (P < 0.001), and tear film breakup time levels (P < 0.001). There was also a significant increase in fluorescein staining scores (P < 0.001) in the model group. Histological examination revealed distinct differences of the corneal epithelium between groups. The corneal epithelium of the model group appeared thicker, with disorganized cell arrangement in the superficial and basal layers, partial defects or detachment of superficial epithelial cells, and a rough, uneven surface. Scanning electron microscopy observations showed a rough corneal epithelial surface with numerous cracks and scattered vesicular-like structures in the model group. Furthermore, the model group rats exhibited a significant increase in expression of IL-1β and TNF-α in tears (P < 0.001), and upregulated expression levels of MMP-9, TNF-α, IL-1β, caspase-3, IL-6, and IFN-γ at both the mRNA and protein levels in corneal tissues (P < 0.001). In conclusion, the modified “wire-meshing cylindrical board” model effectively overcomes the limitations of the traditional “jogging board " dry eye model and successfully simulates the etiology of prolonged visual fatigue. This innovative EDE model demonstrates a high degree of relevance to dry eye conditions resulting from prolonged visual tasks, with a high success rate of model induction. Moreover, it proves to be a simple, practical, and easily replicable model, making it highly suitable for further studies on prolonged visual fatigue and facilitating its widespread adoption in research and clinical applications.

Histological and ultrastructural characterization of the dorso-ventral skin of the juvenile and the adult starry puffer fish (Arothron stellatus, Anonymous 1798)

BackgroundThe starry puffer fish (Arothron stellatus, Anonymous, 1798) is a poisonous tetradontidae fish inhabiting the Red sea. The skin constitutes an important defense against any external effects. The study aims to characterize the dorso-ventral skin of the juvenile and the adult starry puffer fish using light and scanning electron microscopies. Twenty specimens of juvenile and adult fresh fishes were used.ResultsThe scanning electron microarchitecture of the skin of the juvenile and adult fish showed delicate irregular-shaped protrusions, and well-defined bricks-like elevations on the dorsal side and interrupted folds as well as irregular-shaped protrusions on the ventral side. In adult fish, the patterned microridges of the superficial and deep epithelial cells (keratinocytes) were larger and well-defined in the dorsal skin than in the ventral side, the contrary was seen in the juvenile fish. The microridges were arranged in a fingerprint or honeycomb patterns. The openings of the mucous cells were more numerous in the dorsal skin in both age stages but more noticeable in adult. Furthermore, the sensory cells were more dominant in the juveniles than the adults. The odontic spines were only seen in adult. Histologically, few taste buds were observed in the epidermis of the dorsal skin surface of the adult fish. Both mucous and club cells were embedded in the epidermis of the juvenile and adult fish with different shapes and sizes. Melanophores were observed at the dorsal skin of both juvenile and adult fishes while fewer numbers were noticed at the ventral surfaces. Several dermal bony plates with different shapes and sizes were demonstrated in the skin of both adult and juvenile fishes.ConclusionThe structural variations of skin of the juvenile and adult fishes may reflect the various environmental difficulties that they confront.

Suprabasal cells retain progenitor cell identity programs in eosinophilic esophagitis-driven basal cell hyperplasia.

Eosinophilic esophagitis (EoE) is an esophageal immune-mediated disease characterized by eosinophilic inflammation and epithelial remodeling, including basal cell hyperplasia (BCH). Although BCH is known to correlate with disease severity and with persistent symptoms in patients in histological remission, the molecular processes driving BCH remain poorly defined. Here, we demonstrate that BCH is predominantly characterized by an expansion of nonproliferative suprabasal cells that are still committed to early differentiation. Furthermore, we discovered that suprabasal and superficial esophageal epithelial cells retain progenitor identity programs in EoE, evidenced by increased quiescent cell identity scoring and the enrichment of signaling pathways regulating stem cell pluripotency. Enrichment and trajectory analyses identified SOX2 and KLF5 as potential drivers of the increased quiescent identity and epithelial remodeling observed in EoE. Notably, these alterations were not observed in gastroesophageal reflux disease. These findings provide additional insights into the differentiation process in EoE and highlight the distinct characteristics of suprabasal and superficial esophageal epithelial cells in the disease.

Characterization of 3D organotypic epithelial tissues reveals tonsil-specific differences in tonic interferon signaling.

Three-dimensional (3D) culturing techniques can recapitulate the stratified nature of multicellular epithelial tissues. Organotypic 3D epithelial tissue culture methods have several applications, including the study of tissue development and function, drug discovery and toxicity testing, host-pathogen interactions, and the development of tissue-engineered constructs for use in regenerative medicine. We grew 3D organotypic epithelial tissues from foreskin, cervix, and tonsil-derived primary cells and characterized the transcriptome of these in vitro tissue equivalents. Using the same 3D culturing method, all three tissues yielded stratified squamous epithelium, validated histologically using basal and superficial epithelial cell markers. The goal of this study was to use RNA-seq to compare gene expression patterns in these three types of epithelial tissues to gain a better understanding of the molecular mechanisms underlying their function and identify potential therapeutic targets for various diseases. Functional profiling by over-representation and gene set enrichment analysis revealed tissue-specific differences: i.e., cutaneous homeostasis and lipid metabolism in foreskin, extracellular matrix remodeling in cervix, and baseline innate immune differences in tonsil. Specifically, tonsillar epithelia may play an active role in shaping the immune microenvironment of the tonsil balancing inflammation and immune responses in the face of constant exposure to microbial insults. Overall, these data serve as a resource, with gene sets made available for the research community to explore, and as a foundation for understanding the epithelial heterogeneity and how it may impact their in vitro use. An online resource is available to investigate these data (https://viz.datascience.arizona.edu/3DEpiEx/).

Canine Sertoli cell tumor: anti-Müllerian hormone, inhibin B, and estrone sulphate

A unilaterally castrated male boxer was referred for chronic generalized alopecia and pruritus. Physical examination revealed symmetrical alopecia, pendulous prepuce, lichenification of the scrotum, and enlargement of mammary papillae. Penile/preputial cytology revealed superficial epithelial cells. Transabdominal ultrasonographic examination revealed a globoid mass with heterogenous echogenicity in the left caudal abdomen. The presumptive diagnosis was Sertoli cell tumor (SCT) associated with cryptorchidism. Exploratory laparotomy and histology of the removed mass confirmed the diagnosis. Three weeks after surgery, serum anti-Müllerian hormone (AMH) concentrations decreased from 8,435 to 56 ng/ml and inhibin B decreased from 805 to &lt; 6 pg/ml. Two months after surgery dermatoses subsided and there was substantial regression of enlarged nipples. This report highlights the diagnostic value of practical procedures (penile/preputial cytology, transabdominal ultrasonography, and measurement of serum AMH and inhibin B concentrations) to aid in the diagnosis of cryptorchidism and SCT, especially in patients with generalized skin conditions.

Assessment of Cytological Changes in the Oral Mucosa in Young Hematological Patients Treated with Systemic Chemotherapy

The primary objective of the undertaken study was to determine the morphological changes that occur within the oral epithelium in children undergoing chemotherapy following a diagnosis of hematological malignancies. The study group consisted of 18 patients diagnosed with leukemia or lymphoma undergoing treatment with chemotherapy. Swabs (liquid-based cytology) were collected from the oral cavity for microscopic evaluation at baseline, during the chemotherapy cycle with oral mucositis symptoms present, and upon completion of the cycle. Both the neutrophil count and oral mucositis (OM) were registered using the WHO (World Health Organization) scale. The control group included 41 children who were generally healthy. All samples underwent microscopical analyses at the Department of Pathology, Medical University of Gdansk, Poland. A total of 190 cytological preparations were evaluated. The baseline preparations revealed similar cytological images, and the superficial cells of the epithelial layers were seen. A significant (p < 0.01) increase in the number of cells in the intermediate layer of the oral epithelium, as well as a decrease (p < 0.01) in the volume of cells in the superficial layers, was observed in further stages of cytostatic treatment. A decrease in the percentage of superficial epithelial cells with a corresponding increase in the number of intermediate epithelial cells is considered to be a result of toxic damage to the oral mucosa during chemotherapy.

Uropathogenic Escherichia coli biofilms

Urinary tract infection (UTI) is one of the most common infectious diseases, with a global annual incidence of ~175 million cases. Uropathogenic Escherichia coli (UPEC) is the major cause of UTI (&gt;80%) and increasingly associated with rising antibiotic resistance. UPEC form biofilms during infection of the urinary tract, either on the luminal surface of the bladder, intracellularly within bladder superficial epithelial cells, or on the surface of indwelling catheters. This lifestyle of sessile growth promotes enhanced resistance, persistence and increased rates of recurrent UTI. UPEC employ a range of virulence factors to form biofilms, including fimbrial adhesins for attachment and autotransporters to promote cell-to-cell aggregation. In addition, UPEC biofilms are encased in an extracellular matrix comprised of proteins such as curli amyloid fibres and polysaccharides such as cellulose, which together form a hydrating glue that provides structural support for the biofilm and protects its component cells. Here, we describe the key features of UPEC biofilms and their importance for UPEC pathogenesis of the urinary tract.

The potential therapeutic efficacy of intravenous versus subconjunctival mesenchymal stem cells on experimentally ultraviolet-induced corneal injury in adult male albino rats.

The X-rays and the visible light are the main source of ultraviolet radiation (UVR). About 90% of ultraviolet B (UVB) is absorbed by the cornea which may promote corneal inflammation, oedema and damage of its epithelial layer. Bone marrow mesenchymal stem cells (BM-MSCs) have been demonstrated to ameliorate the injured corneal tissue and accelerate its wound healing. This study aimed to compare the healing effect of intravenous (IV) versus subconjunctival (SC) BM-MSCs on the rats' corneas subjected to UVB-irradiation. Ten rats were used as donors for BM-MSCs and the other 40 were allocated into four equal groups: group I (control group), group II (ultraviolet-irradiated group), group III (ultraviolet-irradiated + IV BM-MSCs-treated group) and group IV (ultraviolet-irradiated +SC BM-MSCs-treated group). Rats of all groups were euthanized after 3 weeks and the corneal specimens were processed for histopathological, immunohistochemical and electron microscopy assessment. Ultraviolet-irradiated group showed remarkable thinning of epithelial thickness, wide partial epithelial separation, and desquamation. Neovascularisation of the disorganised stroma and disrupted Descemet's membrane were observed. The superficial and basal epithelial cells appeared irregular and separated by wide intercellular spaces and inflammatory cells. Immunohistochemical examination showed a significant decrease in proliferating cell nuclear antigen immunoreaction. In contrast, minimal changes were observed in rats treated with BM-MSCs with more improvement associated with the subconjunctival administration compared to IV route. Local SC injection of BM-MSCs has an amazing regenerative efficacy on the corneal injury compared to the systemic IV route.

Comparative study of two re-embedding methods on the ultrastructure of corneal tissue

PurposeTo expand the routine of pathological diagnostics of surgical keratoplasty specimens via transmission electron microscopy. The target was to identify the best re-embedding method for optimal structural preservation of formalin fixed paraffin embedded (FFPE) corneal tissue re-embedded into resin for ultrastructural analysis. Basic proceduresBovine FFPE corneal tissue was re-embedded into resin with either a rapid osmium-free four-hour-method or a four-day-routine-method known from nephropathology, compared with primary resin embedded bovine corneal tissue. The analysis involved the ultrastructure of cytoplasm, the intercellular interfaces of superficial epithelial cells, deepest basal epithelial cells and corneal endothelial cells, cell matrix interfaces, Bowman layer, corneal stroma, its microfibril bundles and Descemet membrane. Main findingsThe main observation was the equally reduced preservation status of re-embedded FFPE corneal tissue independent of the used re-embedding method. This extends to the intercellular contacts of superficial epithelial cells and the apical tight junctions of corneal endothelial cells. Hemidesmosomal cell matrix contacts showed less demarcation in re-embedded specimens. Cell matrix interfaces of Bowman layer and Descemet membrane were more clearly bordered in primary resin embedded than re-embedded tissue. In contrast, gap junctions in re-embedded tissue were detected in deepest basal epithelial cells and corneal endothelial cells with comparable preservation to primary resin embedding. Bowman layer, corneal stromal extracellular matrix, its microfibril bundles and Descemet membrane showed equal ultrastructural preservation in all evaluated methods. Principal conclusionCorneal tissue can be successfully analysed with transmission electron microscopy after a rapid osmium-free four hour re-embedding procedure from FFPE material. A comparable morphology with primary resin embedded material can be obtained for gap junctions of deepest basal epithelial cells and corneal endothelial cells, further for Bowman layer, corneal stromal extracellular matrix, its microfibril bundles and Descemet membrane.

P469 Feline aspergillosis caused by Aspergillus candidus: A case report

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMAspergillosis is an emerging mycosis that can occur in apparently healthy animals under environmental stress or immune-compromised condition. In cats, the clinical forms described are sinonasal, synoorbital, and, less frequently, invasive and disseminated forms.Feline aspergillosis is caused mainly by cryptic species of Aspergillus closely related to Aspergillus fumigatus. It is rare to recover species such as A. candidus, an endophytic fungus, opportunistic pathogen, and contaminant of cereals, nuts, and flour. It is described to cause allergic alveolitis or asthma, mycotoxicosis, and other related diseases. Mycelial fragments, inhaled spores, and substances such as terpenoids, flavones, and indoles in the lungs could be involved in the underlying pathogenesis.This presentation aims to describe a clinical case of feline pneumonia caused by A. candidus.Clinical caseWe present the clinical case of a common European breed feline of ∼ 7 years of age. The cat was rescued from the streets of an urban area in Buenos Aires and examined in April 2021. The feline suffered from mild hyperthermia, 39.4°C, had pale to slightly purple mucous membranes, cachexia, dehydration, dyspnea (abdominal respiration), tachycardia, and a large number of parasites. There were no nasal discharge, cough, or skin lesions. A hemogram with a blood biochemistry profile showed no significant alterations. Chest X-rays showed a pattern compatible with severe miliary pneumonia. A bronchoalveolar lavage (BAL) was performed for cytology and microbiological culture; direct examination showed nucleated superficial epithelial cells without atypia, few macrophages, no leukocytes, and gram-positive cocci, while ZN and mycological direct examination were negative. Treatment with doxycycline 10 mg/k every 12 h and dexamethasone was indicated. Flea treatment and control were conducted every 72 h. During the following consultations, the cat continued tachypnoeic and showed increased vesicular murmur with rales and cough. Corticosteroids (Fluticasone propionate 125 mg) were given every 12 h intranasal. Oral parasitization was performed due to the presence of Dipylidium caninum in feces. After 7 days of incubation, white cottony colonies phenotypically identified as Aspergillus grew on Sabouraud dextrose agar. Even though serum galactomannan GM (Platelia Aspergillus EIA BIORAD®) is useful as a biomarker in humans with OD >0.5 positive for invasive aspergillosis, it was used in this case and the result was OD 0.75. Antifungal treatment was started with itraconazole orally at 5 mg/kg/d. A mycobacterial culture was negative.The isolate was identified by partial sequencing of the calmodulin gene CaM as A. candidus.The feline showed a remarkable recovery. After 35 days from the start of the treatment, the final consultation showed a decrease in the pattern of military pneumonia through radiological images and an absence of any evident respiratory symptoms.ConclusionWe describe a fungal infection in a cat caused by an unusual species of Aspergillus. Moreover, this clinical case was proved to be pneumonia. Given this, it is important to consider this species as an emerging pathogen in cats.

Effect of Xylene Oral Exposure on the Mouse Uterus – A Preliminary Study

Abstract Xylene is one of the environmental pollutants with a negative impact mainly on several organ systems. The aim of this study was to determine the effect of xylene on the uterus of mice. The study was performed on 12 adult female mice. Control mice (n = 6) were fed shredded pellets at a dose of 4 g per day. Xylene mice (n = 6) were fed the same diet at the same dose and orally administered xylene at 10 μl per day for 14 days. The mice were synchronized using the Whit-ten effect and introduced to males before the end of the procedure. Mice of both groups with no copulation plug were euthanized by cervical dislocation. The uteri were collected for routine histological and immunohistochemical analysis. The endometrial epithelium demonstrated vacuolar degeneration, mitotic cell activity, and the presence of leukocytes typical of metoestrus. Reductions of the endometrium, stroma, and myometrium were observed in the xylene mice. The xylene application did not have a significant effect on the superficial epithelium, or the size and number of uterine tubular glands. The immunohistochemical analysis of a proliferation marker PCNA revealed that the xylene increased its expression in the stroma, endometrial and myometrial cells, but did not significantly affect the superficial epithelial cells. The expression of an anti-apoptotic marker Bcl-xl in the xylene mice was stronger in the superficial epithelial, stromal, and endometrial cells. The Bcl-xl expression in the myometrial cells was similar to the controls. The results showed that the application of xylene stimulated the proliferation and exerted an anti-apoptotic effect on the uterine cells. However, the increased proliferation can lead to the malignant transformation of cells, resulting in their uncontrollable division.

Effect of smoking on MUC1 expression in oral epithelial dysplasia, oral cancer, and irradiated oral epithelium

ObjectivesThe aim of this study was to assess the MUC1 expression in the oral epithelium of normal, oral epithelial dysplasia (OED), oral squamous cell carcinoma (OSCC), and irradiated oral epithelium (IROE) and its association with smoking habits in non-smokers and smokers. DesignOral mucosal biopsies from controls, OED, OSCC, and IROE groups were obtained and categorized based on the smoking history as non-smokers, smoker I (25 pack-years), and smoker II (>25 pack-years). Immunohistochemical staining of MUC1 using human milk fat globule 1 (HMFG 1) antibody was performed, and the MUC1 score was calculated. The relation between MUC1 expression and clinicopathological findings was examined. ResultsMUC1 staining of superficial oral epithelial cells with mild MUC1 score was detected in all control samples. The MUC1 staining extended from superficial to basal cell layer of oral epithelium with the increase in MUC1 score from moderate to strong in OED, OSCC, and IROE, and the difference was significant (p < 0.004, p < 0.002 and p < 0.004, respectively) compared to controls. A positive association between smoking and MUC1 score was observed within groups (p < 0.05). ConclusionThe depolarization of MUC1 protein expression is associated with smoking habits in OED and OSCC. In the IROE, the radiation causes subcellular and molecular changes, observed as altered MUC1 expression and accelerated by smoking, furthermore, complicating the oral mucosal adaptation and progress to radiation-induced lesions as a delayed effect.

Difference in cytological findings of healthy and conjunctivitis/keratoconjunctivitis affected canine eyes between variably experienced evaluators.

Background and Aim:Cytology investigations are a frequent part of ophthalmological examination. We aimed to assess whether the cytological findings of healthy and conjunctivitis/keratoconjunctivitis samples differed based on the evaluator’s experience.Materials and Methods:A study evaluated healthy eyes (n = 40) and eyes affected with keratoconjunctivitis and/or conjunctivitis (n = 28) in dogs. An ophthalmological examination was performed before sampling the eyes using a sterile cotton swab. An evaluator with theoretical experience and one with undergone clinical pathology residency training performed cytology blinded to the clinical findings.Results:In the healthy eyes group, the agreement between the evaluators for cellularity was nonexistent, while that for cell preservation and mucus content was fair. In the affected eyes group, the agreement for cellularity and mucus content was moderate, while that for cell preservation was fair. The inadequate sample rate differed significantly between the two evaluators in the healthy eyes group (p = 0.006) but not in the affected eyes group (p = 0.083). Bacterial presence was detected by both evaluators, and the findings did not differ statistically from the bacteriology results (p = 0.05). Significant variations were noted in the differential cell count; the mean count of the superficial epithelial cells and goblet cells of the healthy eyes group (p < 0.05) and that of the basal/intermediate cells and neutrophils of the affected eyes (p < 0.05) showed significant differences.Conclusion:The evaluator’s experience significantly affected the differential cell count in both the healthy and affected eyes groups. Neutrophil degeneration was not observed by the less experienced evaluator, whereas bacteria were detected equally well by both the evaluators.

Conjunctival epithelial cells resist productive SARS-CoV-2 infection.

SummaryConjunctival epithelial cells, which express viral-entry receptors angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine type 2 (TMPRSS2), constitute the largest exposed epithelium of the ocular surface tissue and may represent a relevant viral-entry route. To address this question, we generated an organotypic air-liquid-interface model of conjunctival epithelium, composed of basal, suprabasal, and superficial epithelial cells, and fibroblasts, which could be maintained successfully up to day 75 of differentiation. Using single-cell RNA sequencing (RNA-seq), with complementary imaging and virological assays, we observed that while all conjunctival cell types were permissive to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome expression, a productive infection did not ensue. The early innate immune response to SARS-CoV-2 infection in conjunctival cells was characterised by a robust autocrine and paracrine NF-κB activity, without activation of antiviral interferon signalling. Collectively, these data enrich our understanding of SARS-CoV-2 infection at the human ocular surface, with potential implications for the design of preventive strategies and conjunctival transplantation.

Cervical Pre-cancers: Biopsy and Immunohistochemistry.

The existence of precursor lesions for invasive cervical cancer has been recognized for more than 50 years. Our understanding of the pathobiology and behavior of cervical cancer precursors has evolved considerably over the past five decades. Furthermore, the terminology used to classify pre-invasive lesions of the cervix has frequently changed. The realization that human papillomavirus (HPV) infections constitute a morphologic continuum has prompted efforts to include them within a single classification system, specifically the squamous intraepithelial lesions (SILs) which have now been embraced by the surgical pathologists. The reduced number of specific pathological categories has made clinical decision-making more straightforward. The generic criteria for SIL have two important histological parameters: Alterations in the density of superficial epithelial cells and superficial squamous atypia. The flat condyloma or cervical intraepithelial neoplasia (CIN) I is generally associated with intermediate and high-risk HPV types as against the low-risk viruses that cause exophytic/papillary growth patterns of condylomas. The diagnosis of low-grade SIL (LSIL) (flat and exophytic condylomas) requires first excluding benign mimics of LSIL and second to confirm the characteristic cytologic atypia. For high-grade SILs (HSILs), the extent and degree of atypia generally exceed the limits of that described in flat or exophytic condylomas (LSILs). Less maturation, abnormal cell differentiation, loss of cell polarity, and increased mitotic index with abnormal mitotic figures occupying increasing thickness of the epithelium define a lesion as CIN II or CIN III. Atypical immature metaplasia associated with inflammation and atrophy is a challenge in cervical biopsy interpretation. Careful attention to the growth pattern of the epithelium, the distribution of the atypia, nuclear spacing, and the degree of anisokaryosis and the presence of enlarged hyperchromatic nuclei help in differentiating a non-neoplastic from a neoplastic process. This chapter describes in depth the diagnostic difficulties in the interpretation of cervical biopsies. It also provides useful criteria in distinguishing benign mimics from true precancerous lesions and the role of biomarkers such as the p16ink4 and Ki-67 in the differential diagnosis of precursor lesions and the reactive and metaplastic epithelium.

Cytological and microbiological evaluation of conjunctiva in camels with and without conjunctivitis.

To describe the cytological analysis of conjunctiva from normal camels and camels with bacterial conjunctivitis. This study was conducted on 7 normal camels and 15 camels affected with conjunctivitis. The affected camels had a history of conjunctivitis with signs including chemosis, blepharospasm, frequent blinking, and mild-to-moderate serous, mucoid, or purulent ocular discharge. Bacterial swabs were collected from the inferior conjunctival sac of the affected eye without topical anesthetics. Conjunctival smears were obtained from the conjunctival surface for cytological analysis. The cellular analysis of ocular smears revealed a higher percentage of basal cells, neutrophils, eosinophils, and macrophages in camels with conjunctivitis compared with normal camels. In contrast to this, smears from normal camels showed an increased percentage of superficial epithelial cells compared with affected camels. The microbiological assessment of conjunctival swabs collected from affected animals identified a bacterial growth of Staphylococcus aureus., Bacillus sp., Streptococcus sp., Enterococcus faecium., Staphylococcus sp., Corynebacterium sp., Coryne pseudotuberculosis., Saprophytica, Enterobacter cloacae, Escherichia coli, Proteus mirabilis, Proteus vulagaris, and Pseudomonas aeruginosa. It was observed that bacterial conjunctivitis in camels was associated with increased percentages of basal epithelial cells, neutrophils, eosinophils, and macrophages compared with normal camels, while normal camels showed an increased percentage of superficial epithelial cells compared with affected camels.

Is it time for a moratorium on the use of benzalkonium chloride in eyedrops?

Is it time for a moratorium on the use of benzalkonium chloride in eyedrops?