Ultrastructural characteristics of the rats laryngeal mucosa and cartilage on the 7 and 14 days of the experimental opioid effect

Background and objective. Our main task was to study the ultrastructural organization of the laryngeal mucosa and cartilage on the seventh and fourteenth day of opioid exposure. Methods. The material for the study were mature, outbred male rats in the number of 32 animals, weighing 80g, aged 4.5 months. Animals were injected intramuscularly with Nalbuphine once a day at the same time (10-11 hours in the morning) for 14 days. The initial dose of Nalbuphine was 8 mg / kg during the first week, 15 mg / kg during the second week of the experimental study. Thus, we created the conditions for chronic opioid exposure. Animals were divided into 3 experimental groups: 1 – control group; 2 - animals that received the drug for 7 days; 3 - animals that received the drug for 14 days). All animals were kept in a vivarium. Animal care, labeling and all other manipulations were carried out taking into account the issues of bioethical norms. Before collecting necropsy material, the animal was removed from the experiment with diethyl ether. The the larynx mucosa and rats cartilage were used as the material for ultrastructural examination. Ultrastructural capsule was prepared according to conventional methods. Results. As a result of experimental opioid exposure at the end of 7 days we found alternative changes in the epithelial cells of different parts of the larynx, dyscirculatory changes in blood vessels of lamina propria and submucosa, as well as the destruction of chondrocytes of hyaline and elastic cartilages. The hemocapillaries of lamina propria and submucosa, especially in the ventricles of the larynx and in the subchondral region, were dilated, overflowing with erythrocytes. In the laryngeal mucosa and submucosal base of the laryngeal ventricles, in addition to the overflow of hemocapillaries with erythrocytes, a moderate accumulation of perivascular transudate was noted. At the end of the 14th day of the experimental opioid effect, a pronounced hemocapillaries injury was found, which was accompanied by an increase in the permeability of the vascular wall. In addition to blood plasma, erythrocytes were visualized in the enlarged lumens of hemocapillaries. The development of degenerative changes of endothelial cells, which were accompanied by inhomogeneous dilation of the tubules of the smooth and granular endoplasmic reticulum, was noted. The main substance of the connective tissue of the mucous membrane and submucosal base, especially around the vessels was impregnated with transudate.

Morphometric characteristics of the mucous membrane and the cartilaginous component of the larynx are normal at different times during the experimental opioid effect and during withdrawal

Background. The problem of uncontrolled use of opioid drugs is extremely relevant based on the data of domestic and world statistics which are covered in the scientific literature. That is why the study of indicators of morphometric characteristics of the laryngeal mucosa under opioid exposure under experimental opioid exposure will be of interest to both morphologists and practical otorhinolaryngologists. Objective: To study the morphometric parameters of the laryngeal mucosa in normal at different times of the experimental opioid effect and its cancellation. Methods. The material of the study were sexually mature, outbred rats - males in the amount of 61 animals, weighing 80 - 135 g, aged 4.5 - 7.5 months. Histological specimens were prepared according to conventional methods. All morphometric studies were performed using primary (unedited) photographs taken on a Meiji MT4300 LE microscope, Canon EOS 550D x100 lens. All statistical calculations were performed using RStudio v. 1.2.5042. Results. Throughout the experiment, the change of morphometric parameters of the laryngeal mucosa with signs of wavy growth and decline was clearly observed at all times. More positive was the dynamics of morphometric parameters after the abolition of the opioid analgesic, which hypothetically suggests the process of recovery of the mucosa, even after prolonged administration of the opioid.

Deep Convolution Neural Network for Laryngeal Cancer Classification on Contact Endoscopy-Narrow Band Imaging

(1) Background: Contact Endoscopy (CE) and Narrow Band Imaging (NBI) are optical imaging modalities that can provide enhanced and magnified visualization of the superficial vascular networks in the laryngeal mucosa. The similarity of vascular structures between benign and malignant lesions causes a challenge in the visual assessment of CE-NBI images. The main objective of this study is to use Deep Convolutional Neural Networks (DCNN) for the automatic classification of CE-NBI images into benign and malignant groups with minimal human intervention. (2) Methods: A pretrained Res-Net50 model combined with the cut-off-layer technique was selected as the DCNN architecture. A dataset of 8181 CE-NBI images was used during the fine-tuning process in three experiments where several models were generated and validated. The accuracy, sensitivity, and specificity were calculated as the performance metrics in each validation and testing scenario. (3) Results: Out of a total of 72 trained and tested models in all experiments, Model 5 showed high performance. This model is considerably smaller than the full ResNet50 architecture and achieved the testing accuracy of 0.835 on the unseen data during the last experiment. (4) Conclusion: The proposed fine-tuned ResNet50 model showed a high performance to classify CE-NBI images into the benign and malignant groups and has the potential to be part of an assisted system for automatic laryngeal cancer detection.

Role and mechanism of Glut-1 and H+/K+-ATPase expression in pepsin-induced development of vocal cord leukoplakia

Purpose We investigated the role of Glut-1 and H+/K+-ATPase expression in pepsin-induced development of human vocal cord leukoplakia cells (HVCLCs). Next, we analyzed the relationship between Glut-1 and H+/K+-ATPase expression with the clinicopathological features of laryngeal carcinoma. Methods Glut-1 and H+/K+-ATPase expression levels in HVCLCs were determined after treatment with artificial gastric juice containing pepsin and laryngeal carcinoma tissues. Results Exposure to pepsin-containing artificial gastric juice significantly enhanced the migration and proliferation of VSCLCs in a time-dependent manner. The apoptotic rate of VSCLCs decreased over time after exposure to pepsin and reached a nadir on day 7 (p < 0.01). With increasing duration of exposure to pepsin, the proportion of VSCLCs in G0/G1 phase decreased and the proportions in the S and G2/M phases significantly increased (p < 0.05). After treatment with pepsin-containing artificial gastric juice, RT-PCR and Western blotting showed that the expression of Glut-1 and H+/K+-ATPase α, β significantly increased in HVCLCs compared to in the absence of pepsin (p < 0.05). The expression of Glut-1 and H+/K+-ATPase α, β gradually increased from vocal cord leukoplakia (VLC) to laryngeal carcinoma (p < 0.05). Lentivirus-mediated inhibition of Glut-1 expression in VCL significantly inhibited the cells’ migration and proliferation (p < 0.05) but enhanced their apoptosis (p < 0.05). Also, inhibition of Glut-1 expression resulted in an increased proportion of cells in G0/G1 phase and a significantly decreased proportion in G2/M phase (p < 0.05). Conclusions Elevated Glut-1 expression may promote the development of VCL by upregulating laryngeal H+/K+-ATPase expression to reactivate absorbed pepsin, thus damaging the laryngeal mucosa.

Dynamics of changes in the ultrastructural characteristics of the mucosa and cartilage of the larynx of rats at the end of 35 and 42 days of experimental opioid exposure and after 56 days with opioid withdrawal

In our work, we studied the ultrastructural organization of rat laryngeal mucosa and cartilage at the end of 35 and 42 days of experimental opioid exposure and at the end of 56 days with opioid withdrawal. As a result of our study at the end of the 35th day at the ultrastructural level revealed hypersecretion of goblet cells, mucus contained impurities of lymphocytes, single neutrophils and erythrocytes. In the enlarged lumens of the hemocapillaries of the lamina propria of the mucous membrane and submucosal base of different parts of the larynx, the adhesion of erythrocytes was noted. At the end of 42 days in rats, the changes were characterized by moderate damage to cytoplasmic organelles in epitheliocytes. Alternative changes developed in the chondrocytes of elastic and hyaline cartilages. Marked by an increase in the number of active fibroblasts, next to which were numerous fibrils of collagen fibers. The main substance of connective tissue, near dilated vessels, was impregnated with transudate, which was accompanied by the development of perivasal edema. Glycogen grains and vacuoles filled with neutral fats were visualized in the cytoplasm of chondrocytes.At the end of 56 days in rats with two-week withdrawal of the opioid analgesic, we found the development of alternative processes in the epitheliocytes of different parts of the larynx, as well as in chondrocytes, especially in elastic cartilage. Dyscirculatory processes in vessels of own plate of a mucous membrane and a submucous basis of various departments of a larynx were registered. Necrotic changes developed in the columnar epitheliocytes of the submucosal mucosa. Mitochondria swelled, their matrix brightened, cristae collapsed. The results of the article correspond to the research plan of Danylo Halytskyi Lviv National Medical University and are part of the research topic of the Department of Normal Anatomy "Morphofunctional features of organs in pre - and postnatal periods of ontogenesis, under the influence of opioids, food supplements, reconstructive surgery and obesity" 0120U002129) during 2020 - 2024.

A Case of Laryngeal Mucosa Associated Lymphoid Tissue Lymphoma Mimicking Laryngeal Amyloidosis

Laryngeal amyloidosis is a rare disorder characterized by localized amyloid deposition and carries an excellent prognosis. On the other hand, mucosa associated lymphoid tissue (MALT) lymphoma is the most common type of marginal zone lymphoma. MALT lymphoma has a poorer prognosis compared to laryngeal amyloidosis due to more frequent local recurrence, distant metastasis and malignant changes. Despite the differences, as both are found in the form of benign mass and have slow progression, accurate diagnosis is necessary. This report deals with the case of a patient with voice change accompanied by a mass, in which the initial biopsy did not include enough specimen for an accurate diagnosis. However, in the second attempt, an adequate sample was obtained, hence the final diagnosis of laryngeal MALT lymphoma. Based on our recent experience and a review of related literature, we report a case of laryngeal MALT lymphoma mimicking laryngeal amyloidosis.

Dynamics of changes in the structural components of the mucosa and cartilage of the larynx of rats at the end of 42 days of experimental opioid exposure and after 56 days with opioid withdrawal

In our work we aimed to study the pathomorphological manifestations among the structural components of the laryngeal mucosa and cartilage in the later stages of the experimental opioid effect. This goal was achieved through the use of histological techniques to visualize the structural components of the laryngeal mucosa and cartilage. Histological sections with a thickness of 5 -7 mm were made. Histological specimens were prepared according to conventional methods using the dye hematoxylin, eosin, azan according to the method of Heidenhain, PAS-reaction according to Mc-Manus and Alcian blue according to Steedman. Microscopic examinations and photographing of the preparations were performed using an MBI-1 microscope and a Nicon D 3100 digital camera.At the end of 42 days of experimental exposure, the epithelial layer of the epiglottis was inhomogeneously thickened, with excessive accumulation on its pharyngeal and apical part of the stratum corneum. Individual epitheliocytes of the pharyngeal part of the epiglottis underwent necrotic changes, the horny substance loosened in places. On the pharyngeal surface of the mucous membrane found a small amount of mucus with impurities of erythrocytes. Dyscirculatory processes were also registered in the animals of the experimental group. In particular, dilatation and overflow of erythrocytes of vessels of own plate of a mucous membrane of an epiglottis, and also polymorphonuclear infiltration of own plate of a mucous membrane were observed.

ULTRASTRUCTURAL CHARACTERISTICS OF THE LARYNGEAL MUCOSA AND CARTILAGE RAT IS NORMAL

In our work, the aim was to conduct a fundamental study of the issue concerning the ultrastructural characteristics of the mucous membrane and cartilaginous corset of the rat larynx in the norm.This information in the future will make it possible to form a base that will be used to compare the components of the mucosa and laryngeal cartilage in the norm with the dynamics of their changes as a result of experimental opioid exposure.

Systematic review and meta-analysis of prognostic microRNA biomarkers for survival outcome in laryngeal squamous cell cancer

Background Laryngeal carcinoma is a primary malignant tumor originating from the laryngeal mucosa, and its pathogenesis is not fully understood. It is a rare type of cancer that shows a downward trend in the 5-year survival rate. In clinical practice, dysregulated microRNAs are often observed in patients with laryngeal cancer. In recent years, an increasing number of studies have confirmed that the strong biomarker potential of microRNAs. We conducted a systematic review and meta-analysis to identify and highlight multiple microRNAs as biomarkers for disease prognosis in patients with laryngeal cancer. Methods We actively searched the systematic reviews in PubMed, Embase, Web of Science and The Cochrane Library to select the studies that met the proposed guidelines. A total of 5307 patients with laryngeal cancer were included in this study to evaluate the association between microRNAs expression levels and patient outcomes. For overall survival in the clinical stage, a hazard ratio (HR) and corresponding 95% confidence interval (CI) are calculated to assess the effect of survival. Results A total of 36 studies on microRNAs and laryngeal cancer recovery were included in this meta-analysis. The selected endpoints for these studies included overall survival (OS) and disease-free survival (DFS).The comorbidities of overexpression and underexpression of microRNAs were 1.13 (95% CI 1.06–1.20, P < 0.05) and 1.10 (95% CI 1.00–1.20, P < 0.05), respectively. Conclusion MiRNA-100, miRNA-155, miRNA-21, miRNA-34a, miRNA-195 and miR-let-7 are expected to be potential noninvasive and simple markers for laryngeal cancer.