MORPHOLOGICAL AND FUNCTIONAL FEATURES OF THE MUCOUS MEMBRANE OF SMALL AND LARGE INTESTINE IN PATIENTS WITH COVID-19 AND IN POST-COVID-19 PERIOD.

OBJECTIVE: The aim: To reveal the morphological and functional features of the mucous membrane of small and large intestine in patients with COVID-19 and in post-COVID-19 period. PATIENTS AND METHODS: Materials and methods: In the present study, the authors used biopsy and autopsy material represented by the fragments of the mucous membrane of small and large intestine. All studied material was divided into 10 groups. Group 1 (comparison group) included autopsy material from the deceased who did not have COVID-19 during their lifetime. Groups 2-4 included autopsy material from the deceased who had COVID-19 of varying severity during their lifetime. Groups 5-7 included biopsy material from patients who had recovered from COVID-19 of varying severity, while the duration of the post-COVID period ranged from 1 to 50 days. Groups 8-10 included biopsy material from patients who had in anamnesis COVID-19 of varying severity (the duration of the post-COVID period lasted from 51 to 100 days). Histological, immunohistochemical, morphometric and statistical research methods were used. RESULTS: Results: The comparative analysis showed a more expressed deficiency of ACE2 in the mucous membrane of small and large intestine in patients with moderate and severe COVID-19 compared with patients in post-COVID-19 period of different duration. In patients who had moderate and severe COVID-19 in anamnesis, ACE2 deficiency decreases with increasing duration of post-COVID-19 period. In patients recovered from mild COVID-19, the ACE2 content increases with the duration of post-COVID-19 period from 1 to 50 days and corresponds to the norm with the duration of this period from 51 to 100 days. CONCLUSION: Conclusions: The comprehensive morphological study conducted by the authors made it possible, firstly, to clarify the morphological and functional features of the mucous membrane of small and large intestine in patients with COVID-19 of various degrees of severity; secondly, to obtain new data about the morpho-functional state of the mucous membrane of small and large intestine in patients, taking into account different duration of the post-COVID-19 period and the severity of the infection.

RHINOSINUSITIS IN PATIENTS IN POST-COVID-19 PERIOD: ETIOLOGY, CLINICAL AND MORPHOLOGICAL CHARACTERISTICS.

OBJECTIVE: The aim is to identify the etiology, clinical and morphological features of rhinosinusitis in patients in post-COVID-19 period. PATIENTS AND METHODS: 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: 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). CONCLUSION: 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.

SEBORRHEIC KERATOSIS: CURRENT STATE OF THE PROBLEM.

OBJECTIVE: The aim is to identify age, gender, clinical and morphological features of seborrheic keratosis. PATIENTS AND METHODS: Material and methods: The study used biopsy material from 196 patients with a clinical diagnosis of "Seborrheic keratosis". In all cases, when studying directions for pathohistological examination, the authors analyzed gender and age characteristics, as well as localization of seborrheic keratosis. The resulting material was fixed in a 10% solution of neutral formalin (ph 7.4) for 24-48 hours. The procedure was carried out according to the generally accepted technique and the material was embedded in paraffin. Sections of 4-5×10-6m thick were made from paraffin blocks for staining with hematoxylin and eosin. The microspecimens were examined using an Olympus BX-41 microscope (Japan). RESULTS: Results: The study revealed a modern feature of seborrheic keratosis -"rejuvenation" of this tumor, as evidenced by its predominant development in patients aged 31 to 50 years. Seborrheic keratosis is more common in men, and is localized mainly on the skin of the face, scalp, neck and back. When diagnosing seborrheic keratosis, the author notes a significant discrepancy between clinical and morphological diagnoses, which actualizes the problem of improving the available clinical research methods and emphasizes the importance of morphological research. Hyperkeratotic and acanthotic histological variants of seborrheic keratosis are the most common. The case of seborrheic keratosis with malignancy and transformation into squamous cell carcinoma, identified by the author, should develop oncological alertness in doctors and patients. CONCLUSION: Conclusions: The study revealed age, gender, clinical and morphological features of seborrheic keratosis, which will contribute to a better understanding of this pathology by the doctors of various specialties, and improve the treatment and diagnostic process.

ULTRAMICROSCOPIC CHARACTERISTICS OF ERYTHROCYTES IN INDIVIDUALS WITH BORRELIOSIS, RHEUMATOID ARTHRITIS AND TOXOPLASMOSIS AS A MARKER, COMORBID OR CONCOMITANT PATHOLOGY FOR BABESIOSIS.

OBJECTIVE: The aim is to determine the ultramicroscopic characteristics of erythrocytes in individuals with rheumatoid arthritis, borreliosis or toxoplasmosis as a marker, comorbid or concomitant pathology for babesiosis. PATIENTS AND METHODS: Materials and methods: Blood samples from the patients with revealed borreliosis (Lyme disease) (19 cases), toxoplasmosis (15 cases), rheumatoid arthritis in the stage of exacerbation (10 cases) served as the study material (group 2). In all patients of group 2, positive results for babesiosis were obtained during the polymerase chain reaction. The group of comparative control (group 1) consisted of clinically healthy people (n=31), who underwent the blood cytological examination (light microscopy) preceding the scanning electron microscopy, followed by verification of the results by resources of molecular genetic research (polymerase chain reaction). Scanning electron microscopy was used in this study. RESULTS: Results: In patients with babesiosis and marker, comorbid and concomitant conditions for this disease (rheumatoid arthritis, borreliosis, toxoplasmosis) it was identified the specific diagnostic criteria for the presence of extraerythrocyte forms of babesia, constant number of erythrocytes and their regenerative forms, the appearance of degenerative forms of erythrocytes with their size and shape pathology. The latter lead to hemodynamic disorders, the development of ischemic and hypoxic changes in tissues of different organs of human body. CONCLUSION: Conclusions: Scanning electron microscopy of erythrocytes in patients with babesiosis and marker, comorbid and concomitant conditions for this disease (rheumatoid arthritis, borreliosis, toxoplasmosis) plays the role of an objective method of verifying the results of previous clinical and laboratory diagnosis. The use of scanning electron microscopy allow us to determine in these patients the specific diagnostic criteria for the presence of extraerythrocyte forms of babesia, constant number of erythrocytes and their regenerative forms, degenerative forms of erythrocytes with their size and shape pathology.

Seroprevalence of babesiosis in immunocompetent and immunocompromised individuals.

Interest in Babesia species is gaining an increasing attention as an emerging tick-borne pathogen. Infection is primarily transmitted through Ixodes ticks, and alternatively by blood transfusions from asymptomatic donors. AIM: The aim of the study was detection of Babesia seroprevalence in different groups of population with the usage of experimental B. divergens whole-cell slide antigen and commercial B. microti immunofluorescence assay substrate slide. MATERIALS AND METHODS: Indirect immunofluorescence assay trial was performed by testing of 145 blood samples of different origins: healthy individuals (60 - blood donors), risk groups (30 - HIV-infected individuals, 30 - Lyme disease patients) and false-positive IFA controls (10 - seropositive rheumatoid arthritis patients, 15 - patients with toxoplasmosis). RESULTS: The study revealed Babesia antibodies to B. divergens (6.9%) and B. microti (3.4%) that were detected with higher (p <0.05) frequency in HIV-infected individuals (26.7%) and in Lyme disease patients (16.7%) than at blood donors (1.7%). Diagnostically significant IgG titres were detected at 23.3% HIV-infected individuals, 13.3% Lyme disease patients and by 1.7% of blood donors and patients with seropositive latent toxoplasmosis. Specific IgM were detected at 20.0% HIV-infected individuals and 13.3% Lyme disease patients. 57.1% of diagnostically significant titres in HIV-infected and Lyme disease patients were represented by IgG and IgM. CONCLUSIONS: Immunofluorescence assay has a limited use in babesiosis: in acute form with negative microscopy or PCR; in chronic, asymptomatic and subclinical form with low level of parasitemia; and in retrospective and epidemiological studies of the population immune structure. Clinicians need to have increased awareness of babesiosis, and further studies are needed to clarify the optimal management of this infection in risk groups (including HIV-infected patients and blood donors).

CULTURAL METHOD IN BABESIOSIS PATHOGENS DIAGNOSIS: CURRENT STATE OF THE PROBLEM.

OBJECTIVE: The aim is to show the effectiveness of the cultural method in the diagnosis of babesiosis. PATIENTS AND METHODS: Materials and methods: The study material was the blood from 10 healthy humans and animals, 30 humans with borreliosis, 41 animals with babesiosis. A cultural research method was used. RESULTS: Results: The use of cultural method contributed to the optimization of the etiopathogenetic diagnosis of babesiosis. The method objectively proved the ability of protozoa (Babesia spp.) to asexual reproduction in vitro on the multicomponent growth medium with erythrocytes; proved the role of erythrocytes as acceptable for Babesia spp. of target cells. Absolute and relative parameters of detection of Babesia spp. by cultural method in the blood samples of the groups of the examined persons/species were different, making on average 19.7% (with the highest rates among the domestic dogs (62.5%) and cattle (20.0%)). Negative result (0%) was identified among the patients with borreliosis. CONCLUSION: Conclusions: Detection of Babesia spp. by the cultural method objectively confirmed its value as an apparent laboratory criterion for the etiopathogenetic diagnosis of babesiosis.

ULTRAMICROSCOPIC ERYTHROCYTES PROFILE AS A COMPONENT OF THE BABESIOSIS PATHOGENESIS.

OBJECTIVE: The aim is to identify the ultramicroscopic features of the erythrocytes as a component of the babesiosis pathogenesis using scanning electronic microscopy. PATIENTS AND METHODS: Materials and methods: Blood samples from 18 domestic dogs with clinically, microscopically, molecularly and genetically confirmed babesiosis served as the study material. The group of comparative control consisted of clinically healthy people (n=31) and domestic dogs (n=6). The method of scanning electron microscopy was used to study the spatial (three-dimensional coordinate system: the plane formed by the axes OX; OY; OZ) pathology of the size and shape of erythrocytes, microrelief of blood cells surfaces, membranopathies (raptures, macromolecules, conglomerates, micropores). The native peripheral blood smears of various origins were performed in accordance with the generally accepted method without fixation, staining/contrast. For this purpose, 0.01 ml of native blood was applied to the degreased surface of the metal platform (S=1 cm2), dried in the air of the laboratory premise, analyzed (× 2000), using the resources of the equipment (SEO-SEM Inspect S50-B; energy dispersive spectrometer AZtecOne with detector X-MaxN20 (Oxford Instruments plc, UC) and power source in the gun: tungsten cathode with thermoelectron emission. This scanning electron microscope made it possible to examine the object under conditions of accelerating voltage of 100 kV. The analysis of the obtained results was performed in comparison. RESULTS: Results: Under conditions of babesiosis development, the erythrocyte link of hematopoiesis responded by the reduction of the number of normocytes (99.0 % in clinically healthy people and domestic dogs, 77.3 % in dogs with babesiosis), increased of the number of regenerative forms of erythrocytes (1% in clinically healthy people and domestic dogs, 1,5 % in dogs with babesiosis), appearanced of degenerative forms of erythrocytes with characteristic pathology of cell size and shape, membranopathies (damage of the integrity of the membrane with the formation of defects). CONCLUSION: Conclusions: Changes of the ultramicroscopic erythrocytes profile is one of the leading component of the babesiosis pathogenesis. The use of scanning electronic microscopy helps to obtain ultramicroscopic data on the presence of extraerythrocytic forms of Babesia on the background of the impossibility of diagnosis of anisochromia with detailing of inclusions in erythrocytes.

MACROMICROSCOPIC ARGUMENTATION OF THE PATHOGENETIC SCENARIO OF BABESIOSIS IN THE COORDINATE SYSTEM «PATHOGEN-CARRIER-RESERVOIR¼.

OBJECTIVE: The aim is to get a thorough argument for the babesiosis pathogenetic scenario in the coordinate system «pathogen (Babesia spp.) - carrier (ticks of the Ixodoidea superfamily of the Ixodidea family) - reservoir (a susceptible organism)¼ with the emphasis on the epizootic/epidemic role of the carrier. PATIENTS AND METHODS: Materials and methods: The macromicroscopic method of research was used in order to maximize the clarification of the babesiosis scenario, its pathogenetic links, the connection of the latter with attacks of active stages of ixodes ticks, types of circulation of ontogenetic forms of Babesia spp. in the body of carriers and their inoculation of the pathogen into an organism susceptible to it. The use of this method helped to strengthen the diagnostic potential of the study, and increase the reliability of the results obtained. Taking this into consideration it was focused on the epizootological/epidemiological aspects of babesiosis, the role and significance of the most vulnerable epizootic link - Ixodes ticks on the body of the vertebrate provider (mammal), poikilomorphism, anisomorphy. The study of the monolithic idiosome and ticks salivary glands were carried out on activated (capable of attack) female individuals aged 2-3 months after molting. Ticks were dissected in a cool (t=4ºC) Ringer's saline solution for arachnids. Ticks and prepared salivary glands were fixed in 12% formalin solution on 0.1 M phosphate buffer (pH=7.0-7.2) at t=4oC for 3 hours, washed with the buffer, and fixed again for 1 hour (t=4oC). To achieve tonicity, sucrose was added to the fixatives and the washing medium. Dehydration occurred due to a battery of alcohols of increasing concentration and absolute acetone. Microspecimens stained with hematoxylin and eosin were studied using an Olympus BX-41 microscope (Japan). RESULTS: Results: Implementation of the leading stages of the babesiosis pathogenetic scenario is focused on the coordinate system «pathogen (Babesia spp.) - carrier (ticks of the Ixodoidea superfamily of the Ixodidea family) - reservoir (a susceptible organism)¼ in which carrier take the leading place. The macromicroscopic specificity of the structure of the ticks (variability: ability to aniso-, poikilomorphism) is an evidence-based criterion for pathogens inoculation to the macroorganism of warm-blooded vertebrates. It determines the features of circulation and organ/cellular locations of Babesia spp. (intestines and its epithelium, hemolymph, gonads, salivary glands). The species belonging of warm blooded vertebrates susceptible to babesiosis pathogens correlates with the species belonging of ticks and determines the tropicity of the latter. The simultaneous implementation of a complex of research procedures with the tick biological material samples is problematic taking into account the physical lack of material, which requires researchers to re-orient the diagnostic vector towards the use of additional methods for babesiosis diagnosing, including in vitro ones. CONCLUSION: Conclusions: In the pathogenetic scenario of babesiosis, the carrier (Ixodes ticks) is the central figure in the epidemic/epizootic coordinate system.

BIOLOGICAL METHOD FOR BABESIOSIS DETECTION: THE UNIFIED VERSION IN VIVO.

OBJECTIVE: The aim is to establish a unified version of the biological method for babesiosis detection in vivo. PATIENTS AND METHODS: Materials and methods: samples (n=257) of biological material of different origin were examined. These included: blood samples from patients (n=6) and cattle (n=15); salivary gland homogenates (n=28) from 147 imagoes of ticks of the family Ixodidae, 32 imagoes of Ixodes ricinus and 115 imagoes of Dermacentor reticulates; spleen homogenates (n=63) from mouse-like rodents (Muridae) of the genera Myodes, Microtus, Apodemus and Sylvaemus. In order to cultivate in vivo Babesiae of the species B. microti, Syrian hamsters were infected with spleen homogenates from mouse-like rodents; for cultivating the B. divergens species Mongolian gerbils and nonlinear white mices were infected with blood samples from patients and cattle and salivary gland homogenates from ixodic ticks. The technology of modeling was based on the group specificity (differences in susceptibility to parasites and in parameters of morbidity) of the animals, involved in the experiment (Syrian hamsters, Mongolian gerbils, nonlinear white mices). RESULTS: Results: Experimental animals were contaminated by means of intraperitoneal inoculation of 0.3 ml samples of biological material (infected with Babesiae). The animals were infected next day following a day of their preinoculation preparation. The marker parameters for the functional state of experimental animals were as follows: preterm death; appearance and development of clinical-laboratory signs of disease (hypo- or adynamia, loss of appetite, inertness/absence of reactogenicity to tactile/acoustical stimulation, postural changes, wetting of fur, pronounced lameness, flatulence, loss of ≥ 25% of body mass) in them; parasitaemia, histodestruction, cellular detritis. Parasitaemia was detected every two days (beginning with day 8 from the moment of inoculation) by reserves of light and luminescent microscopy. In case of the positive result (revealing of haemoparasites with Babesia spp.-like morphological and tinctorial signs) the verification of Babesiae with their more precise specific identification was performed using the technique of polymerase chain reaction (PCR). Preliminary detection of morbidity parameters in each experimental animal with the artificially created immunocompromised state became an obligatory moment of the described experiment. CONCLUSION: Conclusions: The biological method for detecting Babesia spp. in vivo was improved by the author. This result was achieved by using a double reservoir (Syrian hamsters, Mongolian gerbils and nonlinear white mices with an increased level of susceptibility to parasites) followed by the immunocompromise formation. The use of the improved version of biological method increased the total rate of revealing of Babesiae, therewith creating an objective basis for optimizing the available ways of detection and study of Babesiae in vivo.

MORPHOLOGICAL EFFICACY EVALUATION OF GEL WITH CARBON DIOXIDE EXTRACT OF HOPS IN CASE OF COMPLICATED WOUND INFECTION ACNE VULGARIS.

OBJECTIVE: The aim of the study is to reveal in the experiment the morphological features of the infected skin wounds healing, which are a manifestation of acne vulgaris severe and very severe forms, using a gel with carbon dioxide extract of hops. PATIENTS AND METHODS: Materials and methods: An experimental study was carried out on 80 male WAG rats of three months of age. The animals were divided into 9 groups. Group 1 consisted of intact animals (n=6). Group 2 was represented by animals (n=6), which had hair epilation on the dorsal surface of the body in an area of 1 сm2, followed by application of 2 ml of placebo gel to this area. Group 3 included animals (n=6), which were epilated on the dorsal surface of the body in an area of 1 сm2 and applied 2 ml of 1 % gel with carbon dioxide extract of hops. Group 4 included rats (n=6), which were epilated on the dorsal surface of the body in an area of 1 сm2 and simulated thermal damage. Group 5 was represented by 10 rats, who were epilated on the dorsal surface of the body in an area of 1 сm2, simulated thermal damage, followed by applying 2 ml of placebo gel to the wound surface. Group 6 included rats (n=10), who underwent measures similar to group 5, followed by application of 2 ml of 1 % gel with carbon dioxide extract of hops to the wound surface. In group 7, there were 6 rats, which were epilated on the dorsal surface of the body in an area of 1 сm2, thermal damage to the skin with underlying soft tissues was simulated, followed by application to the wound surface the reference strains of Staphylococcus aureus, Streptococcus pyogenes, Proteus vulgaris, Propionibacterium acnes, Escherichia coli, Pseudomonas aeruginosa, Malassezia slooffiae, Malassezia pachydermatis, Candida albicans, Candida parapsilosis. In groups 8 and 9, there were 15 rats each, which underwent measures similar to group 7, followed by applying 2 ml of placebo gel on its surface on the next day after infection of the wound in group 8, and in group 9 - 2 ml of 1 % gel with carbon dioxide extract hops. The material for the study was the skin with underlying soft tissues. It was used histological, histochemical, morphometric and statistical methods. RESULTS: Results: This experimentally created gel with carbon dioxide extract of hops activates separation processes of horny masses from the surface of the epidermis, cleaning the pores of the skin; stimulating the proliferative activity of the epidermis, which is located in the marginal sections of the wound or covers the surface of the regenerate; activating the processes of cleansing the wound from necrotic tissue; activating the growth and maturation of granulation tissue with its subsequent transformation into connective tissue. It has anti-inflammatory, bactericidal and antimycotic effects, normalizing skin microbiocenosis. CONCLUSION: Conclusions: The complex morphological study has showed that gel with carbon dioxide extract of hops is a highly effective drug in treatment of severe and very severe acne vulgaris, characterized by the development of deep and infected wound defects.

Clinical and morphological features of the ovarian brenner tumour: current state of the problem.

OBJECTIVE: Introduction: Ovarian tumours are an actual problem of present-day medicine, being one of the most difficult sections of modern oncology. The majority of ovarian tumours are of epithelial origin. The ovarian Brenner tumour represents a rare epithelial ovarian neoplasm and accounts for 1-2% of all ovarian neoplasms. The aim of the study is to identify clinical and morphological features of ovarian Brenner tumour. PATIENTS AND METHODS: Materials and methods: The material was 5 cases of Brenner ovarian tumour, diagnosed in the study of 4 cases of operational material and 1 case of autopsy observation for the period from 2007 to 2019. Histological and immunohistochemical staining methods were used. The microspecimens were examined on an Olympus BX-41 microscope (Japan). RESULTS: Results: Ovarian Brenner tumour is a rather rare pathology, the histogenesis of which is debatable. Morphological examination is the main method for its diagnosing. Ovarian Brenner tumours developed in women of middle and old age (the average age was 51.8 years). Women with a malignant ovarian Brenner tumour were older than women with a benign variant (the average age in women with a malignant variant was 55.8 years, with a benign variant - 49.3 years). Benign ovarian Brenner tumour occurred more frequently compared with a malignant one. Malignant and benign variants of ovarian Brenner tumour were characterized by a one-sided nature of the lesion with frequent involvement in the pathological process of the left ovary. Clinically, in patients with a benign variant of the Brenner tumour in all cases an abdominal pain syndrome was determined, combined in one case with metrorrhagia. A malignant ovarian Brenner tumour was clinically manifested by severe abdominal pain syndrome, combined in one case with complaints of an increase in the size of the abdomen, and in another case with intoxication syndrome and a clinic of small bowel obstruction. In all cases a malignant ovarian Brenner tumour metastasized to the omentum and in one case also to the small intestine wall. Macroscopically the ovarian Brenner tumour had the form of a node, the dimensions of which were significantly larger for the malignant variant compared with a benign, dense or soft consistency, on the cross section of a whitish-gray or brown color with cysts. A damaged ovary with a malignant variant of Brenner tumour significantly increased in size, while with a benign one, its size did not change or increased slightly. In all cases the malignant and benign variants of ovarian Brenner tumour were combined with various reproductive system organs pathologies (mucinous papillary cystadenoma of the ovary, serous ovarian cyst, ovarian endometriosis, endometrial hyperplasia, cervical nabothian cysts, uterine leiomyoma). CONCLUSION: Conclusions: A study conducted by the authors revealed clinical and morphological features of a rare ovarian tumour - Brenner tumour, which will contribute to a better understanding of this pathology by the doctors of various specialties, and improve the treatment and diagnostic process.