Epithelial Changes Research Articles

Chemical (Alkali) Burn-Induced Neurotrophic Keratitis Model in New Zealand Rabbit Investigated Using Medical Clinical Readouts and In Vivo Confocal Microscopy (IVCM).

Chemical eye injury is an acute emergency that can result in vision loss. Neurotrophic keratitis (NK) is the most common long-term manifestation of chemical injury. NK due to alkali burn affects ocular surface health and is one of its most common causes. Here, we established a rabbit model of corneal alkali burns to evaluate the severity of NK-associated changes. Alkali burns were induced in NZ rabbits by treating the cornea with (i) a 5 mm circular filter paper soaked in 0.75 N NaOH for 10 s (Mild NK) and (ii) trephination using a guarded trephine (5 mm diameter and 150-micron depth), followed by alkali burn, with a 5 mm circular filter paper soaked in 0.75 N NaOH for 10 s (a severe form of NK). Immediately after, the cornea was rinsed with 10 mL of normal saline to remove traces of NaOH. Clinical features were evaluated on Day 0, Day 1, Day 7, Day 15, and Day 21 post-alkali burn using a slit lamp, Pentacam, and anterior segment optical coherence tomography (AS-OCT). NK-like changes in epithelium, sub-basal nerve plexus, and stroma were observed using in vivo confocal microscopy (IVCM), and corneal sensation were measured using an aesthesiometer post alkali injury. After 21 days, pro-inflammatory cytokines were evaluated for inflammation through ELISA. Trephination followed by alkali burn resulted in the loss of epithelial layers (manifested using fluorescein stain), extensive edema, and increased corneal thickness (550 µm compared to 380 µm thickness of control) evaluated through AS-OCT and increased opacity score in alkali-treated rabbit (80 compared to 16 controls). IVCM images showed complete loss of nerve fibers, which failed to regenerate over 30 days, and loss of corneal sensation-conditions associated with NK. Cytokines evaluation of IL6, VEGF, and MMP9 indicated an increased angiogenic and pro-inflammatory milieu compared to the milder form of NK and the control. Using clinical parameters, we demonstrated that the alkali-treated rabbit model depicts features of NK. Using IVCM in the NaOH burn animal model, we demonstrated a complete loss of nerve fibers with poor self-healing capability associated with sub-basal nerve degeneration and compromised corneal sensation. This pre-clinical rabbit model has implications for future pre-clinical research in neurotrophic keratitis.

Phospholipase D Mediates Glutamine-Induced mTORC1 Activation to Promote Porcine Intestinal Epithelial Cell Proliferation

BackgroundThe intestinal epithelium is one of the fastest self-renewal tissues in the body, and glutamine plays a crucial role in providing carbon and nitrogen for biosynthesis. In intestinal homeostasis, phosphorylation-mediated signaling networks that cause altered cell proliferation, differentiation, and metabolic regulation have been observed. However, our understanding of how glutamine affects protein phosphorylation in the intestinal epithelium is limited, and identifying the essential signaling pathways involved in regulating intestinal epithelial cell growth is particularly challenging. ObjectivesThis study aimed to identify the essential proteins and signaling pathways involved in glutamine’s promotion of porcine intestinal epithelial cell proliferation. MethodsPhosphoproteomics was applied to describe the protein phosphorylation landscape under glutamine treatment. Kinase-substrate enrichment analysis was subjected to predict kinase activity and validated by qRT-PCR and Western blotting. Cell Counting Kit-8, glutamine rescue experiment, chloroquine treatment, and 5-fluoro-2-indolyl deschlorohalopemide inhibition assay revealed the possible underlying mechanism of glutamine promoting porcine intestinal epithelial cell proliferation. ResultsIn this study, glutamine starvation was found to significantly suppress the proliferation of intestinal epithelial cells and change phosphoproteomic profiles with 575 downregulated sites and 321 upregulated sites. Interestingly, phosphorylation of eukaryotic initiation factor 4E-binding protein 1 at position Threonine70 was decreased, which is a crucial downstream of the mechanistic target of rapamycin complex 1 (mTORC1) pathway. Further studies showed that glutamine supplementation rescued cell proliferation and mTORC1 activity, dependent on lysosomal function and phospholipase D activation. ConclusionIn conclusion, glutamine activates mTORC1 signaling dependent on phospholipase D and a functional lysosome to promote intestinal epithelial cell proliferation. This discovery provides new insight into regulating the homeostasis of the intestinal epithelium, particularly in pig production.

A74 A RARE CASE OF ANTIDEPRESSANT-INDUCED LYMPHOCYTIC COLITIS IN A TEENAGER

Abstract Background The etiopathogenesis of microscopic colitis may reflect an immunological reaction in genetically predisposed individuals exposed to an external stimulus causing gut microbiota disruption. Numerous factors such as smoking, autoimmune diseases, and medications have been linked to this condition in adults. Steroids may be required as therapy. Pediatric cases of microscopic colitis have only rarely been reported. Aims Case report. Methods Retrospective single chart review and literature review of microscopic colitis in children. Results A 17-year-old female newly diagnosed with major depression disorder following intentional acetaminophen intoxication was started on sertraline, a selective serotonin reuptake inhibitor (SSRI). Within 2 months from initiation, she began to have non bloody watery diarrhea and had a 5 kg weight loss over a 1-month period . Her laboratory workup only revealed a hypochromic, microcytic anemia. Due to severity of symptoms and reluctance of treating team to discontinue sertraline, she underwent an esophagogastroduodenoscopy and ileo-colonoscopy, both showing grossly normal mucosa. Histological assessment of the colon demonstrated increased intraepithelial lymphocytes, up to 50-60 lymphocytes/100 enterocytes (normal ≤ 20 lymphocytes/100 enterocytes), with associated reactive superficial epithelial changes including goblet cell and mucin depletion, compatible with lymphocytic colitis (Figure1). Sertraline was weaned off, and gastrointestinal symptoms resolved completely. A repeat sigmoidoscopy 5 months later confirmed complete normalization of the histology. Conclusions Microscopic Colitis encompasses two disorders, lymphocytic colitis and collagenous colitis, in which the endoscopic appearance is unremarkable, but histological changes are diagnostic. The clinical manifestations of both are similar and typically include chronic non bloody watery diarrhea, weight loss and abdominal pain. Certain medications have been implicated in causing lymphocytic colitis, including SSRIs. SSRIs historically have not been frequently prescribed in pediatrics, but their use has increased in recent years. The complete resolution of symptoms and histology, without the need for any medical therapy, strongly supports that this lymphocytic colitis was caused by sertraline. The association of lymphocytic colitis with sertraline is not well-known among pediatric practitioners: neither the pediatrician nor psychiatrist in our case were aware of such a possibility. This case highlights that medication adverse events, even if rare, should always be considered when chronology is supportive, and at times, simple discontinuation of the drug is sufficient to completely resolve the issue. Funding Agencies None

A NEW SPECIES OF THELOHANELLUS (CNIDARIA: MYXOSPOREA: MYXOBOLIDAE) FROM THE GILL OF QUILLBACK, CARPIODES CYPRINUS (CYPRINIFORMES: CATOSTOMIDAE), FROM THE ARKANSAS RIVER DRAINAGE OF OKLAHOMA.

During May 2022 and again in March 2023, 5 quillbacks, Carpiodes cyprinus, were collected from the Verdigris River, Wagoner County, Oklahoma (n = 1), and the Black River, Lawrence County, Arkansas (n = 4), and their gill, gallbladder, fins, integument, musculature, and other major organs were macroscopically examined for myxozoans. Gill lamellae from the single quillback from the Verdigris River was infected with a new myxozoan, Thelohanellus oklahomaensis n. sp. Qualitative and quantitative morphological data were obtained from fresh and formalin-fixed preserved myxospores, and molecular data consisted of a 1,767 base pair sequence of the partial small subunit (SSU) ribosomal RNA gene. Phylogenetic analysis grouped T. oklahomaensis n. sp. with myxozoans known to infect North American catostomids and Eurasian cyprinids. Histological examination localized plasmodia to an intralamellar developmental site and revealed a possible vestige of a second polar capsule. Although plasmodia markedly expanded lamellae, there were no associated epithelial or inflammatory changes. Thelohanellus oklahomaensis n. sp. is the only member of the genus known to infect the gills of C. cyprinus.

Naturally occurring phytanic acid exerts anti‐inflammatory effects in a mouse model of dextran sulfate sodium‐induced colitis

AbstractPhytanic acid is one of the branched‐chain fatty acids in the human diet. Humans incorporate it into their bodies through the ingestion of ruminant meat and milk since these animal products contain the 3RS, 7R, 11R‐isomer, as naturally occurring phytanic acid (nPA). Although phytanic acid has been reported to have health‐promoting effects such as immunomodulatory effects, these findings were obtained mostly from in vitro cell‐based experiments. In this study, the in vivo anti‐inflammatory effects of nPA were investigated using a mouse model of dextran sulfate sodium (DSS)‐induced colitis. Mice were fed diets containing 0%, 0.05%, or 0.1% nPA for 4 weeks. During the fourth week, acute colitis was induced by administration of a 3% DSS solution. The intake of 0.05% and 0.1% nPA inhibited DSS‐induced body weight loss in mice, and showed improvement of histopathological changes in colon epithelium. nPA also inhibited the production of proinflammatory cytokines, especially macrophage‐related cytokines such as interleukin‐1β in the colon. These results suggest that nPA exerts anti‐inflammatory effects under in vivo physiological conditions.Practical application: There is a growing demand for fatty acids possessing distinct nutritional and physical characteristics in the realms of health, nutrition, and food‐related applications. The present study demonstrated that a branched‐chain fatty acid, phytanic acid, exerts anti‐inflammatory activity under not only in vitro but also in vivo conditions, using a commonly used mouse model of human inflammatory bowel disease. Our findings encourage further studies to determine whether phytanic acid elicits beneficial properties in humans, and suggest that phytanic acid enrichment can be a novel product strategy in the food market.

Evaluation of the effect of intravitreal injections on corneal epithelial, scleral and limbal region changes in diabetic retinopathy by AS-OCT.

The aim of the study is to evaluate the effects of diabetic retinopathy and intravitreal injections on the corneal, limbal and scleral areas. Patients with diabetes mellitus at different diagnosis and treatment levels were compared among themselves and with the control group in terms of corneal, limbal and scleral aspects with the help of anterior segment optical coherence tomography. In addition, clinical tests such as tear break-up time, Schirmer test-I and ocular surface disease index questionnaire were applied to the patients and the difference between the groups was investigated. When the groups were examined in terms of BUT, SCH-I and OSDI, there was a statistically significant difference between control group and diabetic group(p < 0.05). In the limbal region, all measurements are higher than in patients with diabetic eye involvement. Thinning was detected in the scleral area with intravitreal injection (p < 0.05). It should be known that DM may cause undesirable changes in the limbal region, and the importance of non-invasive detection with AS-OCT should not be forgotten. Since intravitreal injections for DME cause thinning of the sclera, it can cause various complications, and it may be recommended to change the quadrant in repetitive injection applications.

Epithelial overexpression of IL-33 induces eosinophilic esophagitis dependent on IL-13

BackgroundEosinophilic esophagitis (EoE) is an increasingly common inflammatory condition of the esophagus; however, the underlying immunologic mechanisms remain poorly understood. The epithelium-derived cytokine IL-33 is associated with type 2 immune responses and elevated in esophageal biopsies from EoE subjects. ObjectiveWe hypothesized that overexpression of IL-33 by the esophageal epithelium would promote the immunopathology of EoE. MethodsWe evaluated the functional consequences of esophageal epithelial overexpression of a secreted and active form of IL-33 (saIL-33) in a novel transgenic mouse, EoE33. EoE33 mice were analyzed for clinical and immunologic phenotypes. Esophageal contractility was assessed. Epithelial cytokine responses were analyzed in three-dimensional organoids. EoE33 phenotypes were further characterized in ST2-/-, eosinophil-deficient, and IL-13-/- mice. Finally, EoE33 mice were treated with dexamethasone. ResultsEoE33 mice displayed ST2-dependent, EoE-like pathology and failed to thrive. Esophageal tissue remodeling and inflammation included basal zone hyperplasia, eosinophilia, mast cells, and Th2 cells. Marked increases in levels of type 2 cytokines, including IL-13, and molecules associated with immune responses and tissue remodeling were observed. Esophageal organoids suggested reactive epithelial changes. Genetic deletion of IL-13 in EoE33 mice abrogated pathological changes in vivo. EoE33 mice were steroid responsive. ConclusionIL-33 overexpression by the esophageal epithelium generated immunopathology and clinical phenotypes resembling human EoE. IL-33 may play a pivotal role in the etiology of EoE by activating the IL-13 pathway. EoE33 mice are a robust experimental platform for mechanistic investigation and translational discovery.

Gastritis y gastropatías (II). Gastritis crónica atrófica y gastritis hipertrófica

Chronic atrophic gastritis (CAG) is a disease with a multifactorial etiology that produces a depletion of gastric glands, affecting the secretory function of the stomach and the absorption, mainly, of iron and vitamin B12. Its importance lies in the fact that the changes in the gastric epithelium it causes can lead to the onset of gastric cancer. A confirmatory diagnosis is made by means of endoscopic biopsies, which will also determine the need for follow-up on CAG according to the presence or absence of certain histopathological features. There is no specific treatment for CAG beyond the management of its complications.Hypertrophic gastritis can occur as a secondary infiltration of the gastric epithelium or due to pathologies that primarily produce this endoscopic appearance of the gastric mucosa. Of note is Ménétrier's disease, a rare disease whose pathophysiology is largely unknown that is also associated with an increased risk of stomach cancer. It is characterized as a disease that leads to a protein-losing enteropathy. The diagnosis is based on demonstrating the anatomic-pathological changes in the gastric epithelium that are typical of the disease. Treatment involves multiple agents of varying efficacy, such as antisecretory drugs and octreotide.

Type 1 and type 2 torpedo maculopathy.

To analyze torpedo maculopathy (TM) and to report the characteristics of the disease. Retrospective study. The review of a database for clinical diagnosis identified eight patients with TM lesions in the retina between 2016 and 2022. Multimodal imaging was used to analyze the cases. All cases were unilateral, asymptomatic, and hypopigmented. They were associated by surrounding hyperpigmented retinal pigment epithelium changes to varying degrees. All lesions were located in the temporal retina on the horizontal axis, pointing towards the fovea, except for one patient with a lesion inferior to the fovea. Optical coherence tomography imaging revealed a normal inner retina in all eyes. In the area of the TM lesion, attenuation of the interdigitation zone was seen in mild cases (three cases). All other five patients had thinning of the outer nuclear layer and loss of ellipsoid zone and interdigitation zone of the TM lesion. Four of these cases had a subretinal cavitation/cleft, and two of them additionally an inner choroidal excavation. No patient had any sign of choroidal neovascularization. The average age for patients with type 1 TM was 18years and for type 2 TM 16.5years. In this large case series, we could not detect an age difference between the different types of the TM. Contrary to previous discussions, type 2 TM can also occur in young patients.

Single-cell RNA sequencing reveals the epithelial cell, fibroblast, and key gene alterations in chronic rhinosinusitis with nasal polyps

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic inflammatory disease of the nasal mucosa, and epithelial–mesenchymal transition (EMT) is thought to be an essential process in the pathogenesis of CRSwNP. However, the mechanisms of epithelial and fibroblastic changes at the single-cell level are unclear. In this study, we investigated the epithelial cell, fibroblast, and key gene alterations in the development of CRSwNP. We revealed major cell types involved in CRSwNP and nasal mucosal inflammation formation, then mapped epithelial and fibroblast subpopulations. We showed that the apical and glandular epithelial cells and the ADGRB3+ and POSTN+ fibroblasts were the key cell subtypes in the progression of CRSwNP. Pseudotime and cell cycle analysis identified dynamic changes between epithelial cells and fibroblasts during its development. WFDC2 and CCL26 were identified as the key marker genes involved in the development of CRSwNP and were validated by IHC staining, which may provide a potential novel target for future CRSwNP therapy. ScRNA-seq data provided insights into the cellular landscape and the relationship between epithelial cells and fibroblasts in the progression of CRSwNP. WFDC2 and CCL26 were identified as the key genes involved in the development of CRSwNP and may be the potential markers for gene therapy.

UNIQUE MONOCYTE EXPRESSION SIGNATURE IN A STXBP2 MUTATION CAUSING INFANTILE ONSET VEO-IBD AND HLH

Abstract Mutations in syntaxin-binding-protein-2 (STXBP2) can result in NK cell dysfunction leading to a hyperinflammatory state including familial HLH and infantile onset inflammatory bowel disease (IBD). While the function of STXBP2 in NK cells and cytotoxic T cells has been well described, it is highly expressed in monocytes with unclear relevance. We report the case of an infant who presented with severe infantile onset diarrhea with an identified pathogenic mutation in STXBP2. We sought to utilize scRNA-Seq to study the impact of STXBP2 deficiency on monocyte transcription, with the goal of generating insight into the pathogenesis of disease. METHODS: Trio whole exome sequencing was performed. PBMCs were collected from the described patient (case), and two control groups: 3 patients with VEO-IBD, and 3 age matched healthy controls. Single cell RNA-Seq was performed using the 10x platform and data analysis was done using Seurat in R. RESULTS: A male infant presented at 1 week of age with diarrhea and metabolic acidosis. Endoscopic evaluation at 4 weeks of age showed histologic findings of colitis and marked epithelial cell changes, including cell rounding. WES identified a homozygous pathogenic mutation in STXBP2. Due to significant risk of HLH, he underwent evaluation for stem cell transplant. However, in the interim he developed severe HLH requiring ICU level care. He recovered following aggressive treatment and underwent HSCT at 4 months of age and retransplant for graft failure at 11 months of age. He is currently in endoscopic remission off therapy. We analyzed PBMCs from the case patient using single cell RNA-Seq, as compared to patients with VEO-IBD and age-matched controls. STXBP2 expression was most enriched in monocyte populations and reduced in the case patient compared to controls. 231 and 187 genes were differentially expressed in the case monocytes as compared to both controls. The case patient had a paucity of non-classical monocytes. Within the classical monocytes, differentially expressed genes showed enrichment of host defense, cytokine processes, and apoptotic processes. Within the intermediate monocytes, differentially expressed genes showed enrichment of chemotaxis pathways, antigen presentation and T cell activation processes. CONCLUSIONS: Mutations in STXBP2 are a life-threatening cause of fHLH and infantile onset IBD. We report a case of infantile onset IBD due to a homozygous mutation in STXBP2 with resolution of disease post HSCT. Transcriptional analysis of PBMCs from this patient reveals a paucity of non-classical monocytes in the peripheral blood, and differential regulation of genes implicated in host defenses, antigen presentation, chemotaxis and apoptosis. These results may suggest the role of monocyte function in pathogenesis of STXBP2 and in other forms of IBD.

Cytological Features of Oral Cytobrush Smears in Type II Diabetes Mellitus Patients

Oral cytology is a renewed field that aids in diagnosis and observation of possible epithelial changes associated with oral mucosal diseases. Aim; to study the main cytomorphological alteration in gingival and buccal smears from type II diabetics in relation to their hyperglycemic status. The study includes 40 type II diabetic patients (20 new-diagnosed and 20 treated diabetics patients) and 20 healthy persons of both sex. Papanicolaou stained smear were prepared from their cheek and gingiva. The morphological features of 100 unfolded epithelial cells were evaluated under light microscope. Results of this study show that diabetics’ oral mucosa cells characterized by large nuclei with frequent evidence of binucleation, granular chromatin, prominent nucleoli. However, there was frequent small blue cytoplasm and buccal smears showed altered keratinization. As conclusion oral cytology from type II diabetics is associated with detectable cytomorphological changes that is site specific and indicate epithelial cell regeneration and degeneration with altered keratinization especially in buccal mucosa.

Pathology and Monkeypox virus Localization in Tissues From Immunocompromised Patients With Severe or Fatal Mpox.

Pathology and Monkeypox virus (MPXV) tissue tropism in severe and fatal human mpox is not thoroughly described but can help elucidate the disease pathogenesis and the role of coinfections in immunocompromised patients. We analyzed biopsy and autopsy tissues from 22 patients with severe or fatal outcomes to characterize pathology and viral antigen and DNA distribution in tissues by immunohistochemistry and in situ hybridization. Tissue-based testing for coinfections was also performed. Mucocutaneous lesions showed necrotizing and proliferative epithelial changes. Deceased patients with autopsy tissues evaluated had digestive tract lesions, and half had systemic tissue necrosis with thrombotic vasculopathy in lymphoid tissues, lung, or other solid organs. Half also had bronchopneumonia, and one-third had acute lung injury. All cases had MPXV antigen and DNA detected in tissues. Coinfections were identified in 5 of 16 (31%) biopsy and 4 of 6 (67%) autopsy cases. Severe mpox in immunocompromised patients is characterized by extensive viral infection of tissues and viremic dissemination that can progress despite available therapeutics. Digestive tract and lung involvement are common and associated with prominent histopathological and clinical manifestations. Coinfections may complicate mpox diagnosis and treatment. Significant viral DNA (likely correlating to infectious virus) in tissues necessitates enhanced biosafety measures in healthcare and autopsy settings.

Benign Phyllodes Tumor with Extensive Cystic Squamous Metaplasia, Clinically Presenting as Malignant Phyllodes

Abstract Phyllodes tumor are very rare fibroepithelial lesions which comprises 0.3%–1% of all primary breast tumors with peak age around 40–50 years. Phyllodes tumor are classified ino three types: Benign, borderline and malignant tumors. Metaplastic change is very rare in these tumors and can involve both epithelial or stromal components. Stromal metaplastic changes include cartilage, bony or fatty change. Epithelial metaplastic changes include apocrine or squamous change. About 10% of phyllodes tumor show squamous metaplasia. We report a case of 47-year-old patient presenting with large lump in right breast measuring 16 cm × 15 cm × 11 cm, occupying all four quadrants. Clinically, it appeared as malignant phyllodes. But, microscopically, it was benign phyllodes tumor having extensive cystic squamous metaplasia.

Bisphenol A exposure stimulates prostatic fibrosis via exosome-triggered epithelium changes

Fibrosis is the pathological basis for the clinical progression of benign prostatic hyperplasia (BPH). Prostatic fibrosis is an important risk factor in patients with BPH who experience lower urinary tract symptoms. Bisphenol A (BPA) is an environmental endocrine disruptor (EED) that causes prostate defects. The effects of BPA on the prostate were investigated in this study using mouse and human prostate cell models. BPA-induced mouse prostatic fibrosis is characterized by collagen deposition and an increase in hydroxyproline concentration. Furthermore, BPA-exposed prostatic stromal fibroblasts exosomes promote the epithelial-mesenchymal transition of epithelial cells. High-throughput RNA sequencing and functional enrichment analyses show that substantially altered mRNAs, lncRNAs and circRNAs play roles in cellular interactions and the hypoxia-inducible factor-1 signaling pathway. The results showed that exosomes participated in the pro-fibrogenic effects of BPA on the prostate by mediating communication between stromal and epithelial cells and triggering epithelial changes.

Histological evaluation of the surgical margins of oral soft tissue incisions using a dual-wavelength diode laser and an Er, Cr:YSGG laser; an ex vivo study.

This study compared a dual-wavelength diode laser and an Er, Cr:YSGG laser in oral soft tissue incisions to determine the most effective and safest laser system at the histopathological level. The (810 and 980 nm) dual-wavelength diode laser was used at 1.5 W and 2.5 W (CW) power settings, and the (2780 nm) Er, Cr:YSGG laser was used at 2.5 W and 3.5 W (PW) power settings. Both laser systems were used to incise the tissues of freshly dissected sheep tongue pieces to obtain the following histopathological criteria: epithelial tissue changes, connective tissue changes, and lateral thermal damage extent by optical microscopy. The epithelial and connective tissue damage scores were significantly higher in the dual-wavelength diode laser groups than in the Er, Cr:YSGG laser groups (P<0.001), and there was a significant difference between some groups. The extent of lateral thermal damage was also significantly higher in the diode laser groups than in the Er, Cr: YSGG laser groups (P<0.001), and there was a significant difference between groups. Group 2 (2.5 W) of the diode laser was the highest for all three criteria, while group 3 (2.5 W) of the Er, Cr:YSGG laser was the lowest. The Er, Cr:YSGG laser with an output power of 2.5 W is, histologically, the most effective and safest laser for oral soft tissue incision. The dual-wavelength diode laser causes more damage than the Er, Cr:YSGG laser, but it can be used with a low output power and 1 mm safety distance in excisional biopsy.

New possibilities for reparative therapy of dry eye syndrome

This study evaluates the clinical efficacy of the drug Optinol Soft Recovery in the treatment of patients with DES accompanied by xerotic changes in the corneal epithelium. The study included 82 patients (15 children and 65 adults) with moderate and severe DES accompanied by the following corneal pathology: filamentous keratitis (20 patients, 33 eyes), persistent corneal erosion (28 patients, 49 eyes) and punctate keratopathy (34 patients, 68 eyes). All patients receiving fourfold instillations of the studied drug were observed already during the first 7 days to have increased stability of the tear film and decreased severity of staining of the cornea and conjunctiva with vital dyes (0.1% sodium fluorescein and 3% lissamine green, respectively). Further, as the patients were transferred to an individual instillation regimen, a progressive decrease in the Ocular Surface Disease Index (OSDI), an increase in corneal sensitivity and tear meniscus index were also recorded. The differences in most parameters of the course of xerosis compared to the initial ones were statistically significant starting from day 10-20 of therapy, depending on the initial severity of corneal xerosis (p<0.05-0.001). Patients with moderate DES complicated by punctate keratopathy were the most susceptible to therapy with the drug Optinol Soft Recovery, while patients with filamentous keratitis secondary to a severe clinical form of DES were the least susceptible.

Оценка эффективности применения активированной глицирризиновой кислоты у пациенток с ВПЧ-ассоциированными поражениями шейки матки с учетом уровня половых стероидных и гонадотропных пептидных гормонов

Background: the persistence of human papillomavirus (HPV) infection with oncogenic protein expression causes changes in the stratified squamous epithelium, which is the immediate cause of cervical cancer manifestation. The use of the activated glycyrrhizic acid (GA) for HPV-associated cervical lesions is pathogenetically justified. During an experimental study on a culture of human cervical adenocarcinoma cells (HeLa line), it was shown that the antiproliferative effect of GA is enhanced by low concentrations of estradiol and weakened by testosterone. Aim: to evaluate the efficacy of the activated GA in female patients with HPV-associated cervical pathology with respect to the level of sex steroid and gonadotropin-releasing hormones Patients and Methods: a prospective study was conducted with the inclusion of 51 female patients with HPV-associated cervical pathology (ASCUS — atypical squamous epithelial cells of undetermined significance, LSIL — low-grade squamous intraepithelial lesion, HSIL— high-grade squamous intraepithelial lesion according to the Bethesda classification) and the absence of clinical and/or laboratory signs of hyperandrogenism. The following studies were performed: clinical examination, colposcopy, microscopic examination, cytology and immunocytochemistry, as well as hormonal status was evaluated. All female patients received Epigen Intim spray 0.1%, 3 times a day, for 30 days from the moment of inclusion in the study. If indicated, cervical conization was also performed. The treatment efficacy was evaluated 90±7 days after the therapy initiation based on the liquid-based HPV-PAP test results with the determination of p16/Ki67 oncogenic protein co-expression in the cervical epithelium. Changes in the colposcopy results were also evaluated. Results: it was found that the level of estradiol in patients with HSIL was lower versus the patients with ASCUS and LSIL (32 [32.0; 36.0] pg/ml versus 40 [34.5; 45.5] and 40 [33.0; 44.0] pg/ml, respectively, but the difference did not reach statistical significance (p=0.453). 90 days after the follow-up start, according to the cytology results of the cervical epithelium, 50 (98.0%) patients showed the absence of cervical intraepithelial lesions and the absence of p16/Ki67 oncogenic protein co-expression, 48 (94.1%) patients showed no high-risk (HR) HPV DNA, whereas HPV-associated colposcopy signs (anogenital warts, chronic cervicitis signs) persisted only in 6 (11.8%) patients. Conclusion: the use of activated GA in female patients of reproductive age with HPV-associated cervical lesions without clinical and/or laboratory signs of hyperandrogenism allowed to achieve high efficiency indicators: 98.5% — according to the cytology results, 83.3% — in reducing the HR-HPV viral load, 85.7% — in terms of p16/Ki67 oncogenic protein co-expression. KEYWORDS: human papillomavirus, squamous intraepithelial lesion of the cervix, oncogenic protein co-expression, sex steroid hormones, activated glycyrrhizic acid FOR CITATION: Mandrykina Zh.A., Dobrokhotova Yu.E., Ibragimova D.M., Kazieva M.D., Filimonova M.S., Shimanovsky N.L., Akhmetgaliev A.R. Assessment of the activated glycyrrhizic acid efficacy in female patients with HPV-associated cervical lesions with respect to the level of sex steroid and gonadotropin-releasing hormones. Russian Journal of Woman and Child Health. 2024;7(2):127–134 (in Russ.). DOI: 10.32364/2618-8430-2024-7-2-7.

Increased secretory autophagy in retinal pigment epithelial cells of the 5xFAD mice

Purpose: Disturbed proteostasis and oxidative stress have a key role in Alzheimer's disease and age‐related macular degeneration pathology. Constant oxidative stress and disturbed proteostasis lead to intracellular protein aggregation and possibly the activation of secretory autophagy. In this study we assessed the oxidative stress and secretory autophagy related biomarker changes in the retinal pigment epithelium (RPE) of the 5xFAD mice model expressing abundant levels of the human beta‐amyloid.Methods: The eyes of 3‐, 6‐ and 12‐month‐old WT (n = 5) and 5xFAD (n = 5) mice were enucleated, paraffin fixed and examined by immunomicroscopy. The paraffin sections were immunostained for beta‐amyloid and the key secretory autophagy markers Sec22b, TRIM16, galectin 8, IL‐1β, HMGB1 and ferritin. Moreover, the sections were stained for oxidative stress marker 4‐HNE and antioxidant enzyme catalase. Protein degradation related markers ubiquitin (Ub) and SQSTM1/p62 were analysed from 6‐ and 12‐months timepoints.Results: Immunohistochemistry analysis revealed that the oxidative stress and protein degradation related markers were upregulated in the RPE of 6 and 12‐months old 5xFAD mice suggesting presence of oxidative stress and protein aggregation. TRIM16 and galectin 8 were increased in the RPE of the 6‐ and 12‐months old 5xFAD mice, while the increased expression of Sec22b was observed in 3‐ and 6‐months timepoint. IL‐1β, HMGB1 and ferritin were accumulated at the basal side of the RPE cells/Bruch's membrane in 6‐months old 5xFAD mice. The high accumulation of beta‐amyloid was detected in drusen like structures. Electrophysiological analysis revealed decreased scotopic and photopic signalling compared to WT mice.Conclusions: 5xFAD mouse model seems to be a good model to study protein aggregation and activation of the secretory autophagy in response to degenerative retina processes.

The association of pre-photorefractive keratectomy Schirmer-1 test value with postoperative corneal epithelial thickness, ocular surface discomfort, and visual acuity.

To investigate the association of pre-photorefractive keratectomy Schirmer-1 test value with post-photorefractive keratectomy central corneal epithelial thickness, ocular surface disease index score, and uncorrected distance visual acuity. Patients were categorized according to preoperative Schirmer-1 value: the normal Schirmer Group (n=54; Schirmer-1 test value, >10 mm) and the low Schirmer Group (n=52; Schirmer-1 test value, between 6 and 10 mm). We analyzed ablation depth, visual acuity, result of Schirmer-1 test (with anesthesia), tear film break-up time, ocular surface disease index score, central corneal epithelial thickness, and spherical equivalent refraction. We found significant differences between the groups in Schirmer-1 test value, tear film break-up time, and ocular surface disease index score, both preoperatively and postoperatively (p<0.001). The preoperative central corneal epithelial thicknesses of the two groups were similar (p>0.05). After photorefractive keratectomy, the Schirmer-1 test value and spherical equivalent refraction decreased in both groups (p<0.05), and ocular surface disease index scores and central corneal epithelial thickness values increased in the low Schirmer Group (p<0.001) but not in the normal Schirmer Group (p>0.05). The postoperative central corneal epithelial thicknesses of the low Schirmer Group were significantly higher than those of the normal Schirmer Group (p<0.001). Postoperative uncorrected distance visual acuity did not differ significantly between the two groups (p>0.05). In patients with low Schirmer-1 test values before photorefractive keratectomy, the corneal epithelium thickened and ocular surface complaints increased during the postoperative period. However, changes in the corneal epithelium did not affect the postoperative uncorrected distance visual acuity. To reduce postoperative problems on the ocular surface in these patients, we recommend that dry eye be treated before photorefractive keratectomy.