The safety of food additives E407 and E407a has raised concerns in the scientific community. Thus, this study aims to assess the local and systemic toxic effects of the common food additive E407a in rats orally exposed to it for two weeks. Complex evaluations of the effects of semi-refined carrageenan (E407a) on rats upon oral exposure were performed. Local effects of E407a on the intestine were analyzed using routine histological stains and CD68 immunostaining. Furthermore, circulating levels of inflammatory markers were assessed. A fluorescent probe O1O (2- (2′-OH-phenyl)-5-phenyl-1,3-oxazole) was used for evaluating the state of leukocyte cell membranes. Cell death modes of leukocytes were analyzed by flow cytometry using Annexin V and 7-aminoactinomycin D staining. Oral administration of the common food additive E407a was found to be associated with altered small and large intestinal morphology, infiltration of the lamina propria in the small intestine with macrophages (CD68+ cells), high systemic levels of inflammation markers, and changes in the lipid order of the phospholipid bilayer in the cell membranes of leukocytes, alongside the activation of their apoptosis. Our findings suggest that oral exposure to E407a through rats results in the development of intestinal inflammation.
The aim is to identify the etiology, clinical and morphological features of rhinosinusitis in patients in post-COVID-19 period. Materials and methods: In the present study, it was carried out the analysis of 11 cases of rhinosinusitis, which developed after COVID-19 infection. The diagnosis of rhi¬nosinusitis was established on the basis of anamnesis, clinical and laboratory examination, specialized instrumental examination (rhinoendoscopy, X-ray, magnetic resonance imaging, spiral and 3D computed tomography). All patients underwent endoscopic sanitation of the nasal cavity, expansion of the maxillary anastomosis, maxillary sinusotomy, sanitation of the maxillary sinuses and removal of pathologically altered tissues. Microbiological examination of the swab from the nasal cavity was carried out in all patients. Histological and morphometric research methods were used during the morphological study of surgical material. The nonparametric Mann-Whitney U test was used to compare the means in the groups. Results: The conducted comprehensive study made it possible to identify chronic atrophic rhinosinusitis at the stage of exacerbation caused by associations of bacteria and fungi in patients in post-COVID-19 period. Among bacteria, the authors most often noted Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella pneumonia, Streptococcus pneumonia and Enterococcus faecalis. Among fungi, there were Aspergillus, Candida, Mucor and Coccidioides. Fungal infection was characterized by invasion into the mucous membrane of the nose and paranasal sinuses. In patients in post-COVID-19 period the invasive bacterial-fungal chronic atrophic rhinosinusitis at the stage of exacerbation was predominantly bilateral, characterized by the involvement of several or all paranasal sinuses in the process. Patients with such pathology complained of periodic fever, headaches and malaise; nasal congestion and constant difficulty in nasal breathing; yellowish-greenish-reddish discharge from the nasal cavity, sometimes with a fetid odor; discomfort and pain in the area of paranasal sinuses; immobility of the eyeball, hyposmia or anosmia; reduction or complete loss of vision. Frequent risk factors for the development of invasive bacterial-fungal chronic atrophic rhinosinusitis at the stage of exacerbation in patients in post-COVID-19 period were the information about moderate or severe course of this infection in anamnesis; comorbidities (predominantly diabetes mellitus, hypertensive disease and ischemic heart disease). Conclusions: The study conducted by the authors made it possible to identify the etiological, clinical and morphological features, as well as risk factors of rhinosinusitis in patients in post-COVID-19 period. This information will contribute to a better understanding of such pathology by the doctors and improve the diagnostic and treatment process.
Introduction. Rare-earth orthovanadate nanoparticles (ReVO4:Eu3+, Re = Gd, Y or La) are promising agents for photodynamic therapy of cancer due to their modifiable redox properties. However, their toxicity limits their application. Objective. The aim of this research was to elucidate pro-eryptotic effects of GdVO4:Eu3+ and LaVO4:Eu3+ nanoparticles with identification of underlying mechanisms of eryptosis induction and to determine their pharmacological potential in eryptosis-related diseases. Methods. Blood samples (n=9) were incubated for 24 h with 0-10-20-40-80 mg/L GdVO4:Eu3+ or LaVO4:Eu3+ nanoparticles, washed and used to prepare erythrocyte suspensions to analyze the cell membrane scrambling (annexin-V-FITC staining), cell shrinkage (forward scatter signaling), reactive oxygen species (ROS) generation through 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) staining and intracellular Ca2+ levels via FLUO4 AM staining by flow cytometry. Internalization of europium-enabled luminescent GdVO4:Eu3+ and LaVO4:Eu3+ nanoparticles was assessed by confocal laser scanning microscopy. Results. Both nanoparticles triggered eryptosis at concentrations of 80 mg/L. ROS-mediated mechanisms were not involved in rare-earth orthovanadate nanoparticles-induced eryptosis. Elevated cytosolic Ca2+ concentrations were revealed even at subtoxic concentrations of nanoparticles. LaVO4:Eu3+ nanoparticles increased intracellular calcium levels in a more pronounced way compared with GdVO4:Eu3+ nanoparticles. Our data disclose that the small-sized (15 nm) GdVO4:Eu3+ nanoparticles were internalized after a 24 h incubation, while the large-sized (~30 nm) LaVO4:Eu3+ nanoparticles were localized preferentially around erythrocytes. Conclusions. Both internalized GdVO4:Eu3+ and non-internalized LaVO4:Eu3+ nanoparticles (80 mg / L) promote eryptosis of erythrocytes after a 24 h exposure in vitro via Ca2+ signaling without involvement of oxidative stress. Eryptosis is a promising model for assessing nanotoxicity.
The aim was to reveal the morphological features of the lungs in post-COVID-19 syndrome. Materials and methods: The material of the study was autopsy material – fragments of the lung tissue from 96 deceased (59 men and 37 women). During the lifetime, all patients had in anamnesis COVID-19 of varying severity, and after the treatment of this infection, they had various manifestations of respiratory failure until death. The average duration of the post-COVID-19 period was 148.6±9.5 days. Based on the severity of COVID-19 in anamnesis, all cases were divided into three groups. Group 1 included 39 cases with mild COVID-19 in anamnesis. Group 2 included 24 cases with moderate severity of COVID-19 in an¬amnesis. Group 3 included 33 cases with severe COVID-19 in anamnesis. Histological, histochemical, morphometric and statistical research methods were used. Results: Morphological features of the lungs in post-COVID-19 syndrome were the presence of pneumosclerosis; focal-diffuse immune cells infiltration; emphysematous and atelectatic changes; degenerative-desquamatic changes in the alveolar epithelium; metaplastic changes of connective tissue; dystrophic calcification; dystrophic, metaplastic and dysplastic changes in the epithelial layer of bronchial tree; hemodynamic disorders. Pneumosclerosis, focal-diffuse immune cells infiltration, alterative changes in the alveolar epithelium, emphysematous and atelectatic changes, hemodynamic disorders increased with an increase the severity of COVID-19. Metaplastic changes of connective tissue, dystrophic calcification, dystrophic, metaplastic and dysplastic changes in epithelial layer of bronchial tree did not depend on the severity of the infection. Conclusions: The changes identified by the authors help to explain pulmonary manifestations of post-COVID-19 syndrome. They should be the basis for the oncological alertness formation among doctors, the development of rehabilitation and treatment measures for such category of patients.
The aim is to reveal the expression features of MCA to human papilloma virus type 16 and anti-Epstein-Barr virus in the pleomorphic adenoma, surrounding and intact salivary gland. Materials and methods: It was used surgical and biopsy material from 30 patients, represented by pleomorphic adenomas with surrounding to tumor tissue of the salivary gland and intact tissue of the salivary gland (the distance between the tumor and the intact salivary gland – 10 mm). Immunohistochemical study was performed using mouse monoclonal antibody (MCA) to human papilloma virus type 16 (clone CAMVIR-1, «Diagnostic BioSystems», USA) and anti-Epstein-Barr virus (LMP, clone CS. 1-4, «Dako», Denmark). Visualization was performed, using an EnVisionTM FLEX detection system (Dako, Denmark). Antigen unmasking was carried out in citrate buffer pH 6.0 at 95°C. Primary antibodies were incubated at room temperature for 30 minutes, secondary antibodies – 20 minutes. Sections were counterstained with Gill hematoxylin. We assessed the immunohistochemical reaction by a semi-quantitative method by counting the percentage of positively stained cells in the field of view of a microscope × 400. Microspecimens were studied and photoarchived on an Olympus BX-41 microscope (Japan). Results: In this study it was detected a positive immunohistochemical reaction with MCA to human papilloma virus type 16 and anti-Epstein-Barr virus, respectively, in 26 (86.7%) and 8 (26.7%) cases. Epithelial, mixed and mesenchymal variants of pleomorphic adenoma of the salivary glands are characterized, respectively, by the severely expressed, moderately expressed and minimally expressed of MCA to human papilloma virus type 16 and anti-Epstein-Barr virus. The parenchymal component of pleomorphic adenoma is characterized by more marked expression of these markers as compared to the stromal component. The epithelial cells of the salivary glands, surrounding the pleomorphic adenoma, as well as intact salivary glands, express MCA to human papilloma virus type 16 and anti-Epstein-Barr virus. The severity of the expression of these markers in the salivary gland is determined by the histological variant of the tumor (severely expressed in the epithelial variant, moderately expressed in the mixed variant, and minimally expressed in the mesenchymal variant). Conclusions: The immunohistochemical study has shown that the Epstein-Barr virus and, especially, human papilloma virus type 16 can act as exogenous trigger factors involved in the development of pleomorphic adenoma of the salivary glands. The revealed immunohistochemical features of MCA expression to human papilloma virus type 16 and anti-Epstein-Barr virus in the salivary gland surrounding the pleomorphic adenoma and in the intact tissue of the salivary gland make it possible to recommend the extracapsulardissection of the tumor with resection of the adjacent intact tissue of the salivary gland at a distance of 10 mm in patients with pleomorphic adenoma.
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