Neutral Formalin Solution Research Articles

FUNCTIONAL POSSIBILITIES OF ESOPHAGEAL-GASTIC ANASTOMOSIS IN THE REMOTE POSTOPERATIVE PERIOD ACCORDING TO THE DATA OF THE MORPHO-FUNCTIONAL STUDY

Resume. Purpose: Morpho-functional assessment of the method of endoprosthesis of failure of the esophageal-gastric anastomosis sutures in the distant postoperative period. Introduction. Currently, views on treatment tactics in the case of the inability to suture esophageal anastomoses are contradictory and have diametrically opposite opinions — from extremely gentle to extremely aggressive. Optimism of supporters of palliative resections is due not only to the almost identical mortality with radical operations, which is 4.0-31.6%, but also to the possibility of ensuring 5-year survival of 8.5-15.7% of operated patients. Thus, a number of questions remain relevant, including the study of morpho-functional shifts in tissue cells of the esophageal-gastric anastomosis with the assessment of its functional capabilities in the distant postoperative period based on morphological research. Material and methods. The work was performed in the Department of Surgery of the Gastrointestinal Tract and Esophagus of the State University «IZNG named after V.T. Zaitsev National Academy of Medical Sciences of Ukraine» with the presentation of the results of a morpho-functional study and statistical analysis of the treatment of patients with the development of failure of esophageal-intestinal anastomoses in the postoperative period. All patients were fitted with self-expanding stents, stents from M.I.Tech Co., Ltd. (Korea), covered esophageal HANAROSTENT with an antireflux valve - 14 and Boston Scientific Corp. were used for endoprosthetics. (USA), covered with Ultraflex-2 esophagus. In the long-term period, all patients discharged from the hospital underwent a control X-ray and endoscopic examination every 1-2 months, during which a biopsy was performed and material was taken for the next morphological examination. For light microscopy, tissue fragments of the esophageal-gastric junction were fixed in a 10% solution of neutral formalin for 24 hours at a temperature of +20°C, after which they were dehydrated with ethyl alcohol and embedded in paraffin. Sections were prepared from paraffin blocks and stained with Ehrlich’s hematoxylin and eosin. Research results. The long-term results of endoprosthesis of failed esophageal-gastric anastomosis sutures in 43 stented patients were evaluated. Of great interest is the morpho-functional state of the esophageal-gastric anastomosis in the long term after surgery (3- 24 months), as these terms determine the functional capabilities of the anastomosis. Taking into account that during the research it is possible to detect initial morpho-functional shifts in the tissues of the examined organs, it should be assumed that in the future hypoxia of the stomach and esophagus increases and the degree of expressiveness of these disorders maintains the same sequence. Conclusions. The conducted studies expanded the understanding of the pathogenesis of morpho-functional disorders occurring in the distant postoperative period, confirmed the priority of metabolic disorders in tissue cells in diseases of the gastrointestinal tract, and also allowed to give a prognostic assessment of the functional possibilities of the further development of the gastric process. further tactics of managing patients at the next stage of treatment.

Epidemiological and pathomorphologic investigation of peste des petits ruminants in Western Algeria: A comprehensive study of clinical and histopathological findings.

This study delves into the epidemiology and pathomorphologic characteristics of peste des petits ruminants (PPR) in western Algeria, a viral disease that constantly threatens small animals in Africa, the Middle East, and Asia. The purpose of this investigation was to evaluate the epidemiology of PPR in western Algeria and to understand the pathomorphological lesions in naturally infected small ruminants. An online survey conducted via google forms and shared with veterinarians in the wilaya of Tiaret, provided insights into the prevalence and clinical manifestations of PPR.A comprehensive examination of organs was conducted and representative tissue samples from the lungs, trachea, thymus, spleen, liver, kidney, heart, tongue, stomach, different parts of the small and large intestine, and mesenteric lymph nodes were collected and the specimen was fixed in a 10% neutral buffer formalin solution. Among 2,200 small ruminants managed by expert veterinarians, 192 small ruminants exhibited clinical signs compatible with PPR, and 79 dead animals. Among the 31 sick young small ruminants, eight were confirmed to be infected with the PPR virus. Necropsies of affected animals revealed significant gross lesions in organs such as the lungs, intestines, spleen, and lymph nodes. Histopathological analysis further illuminated the severity of lesions, including interstitial pneumonia, syncytial cell formation, and severe gastroenteritis. The study's comprehensive approach, encompassing epidemiological data, necropsy findings, and histopathological insights, contributes valuable knowledge for understanding and managing PPR outbreaks.The pathological lesions observed in this study exhibited consistency with those previously documented in experimental studies, thereby providing support for the diagnosis based on clinical signs and disease history.

Quantitative study of the ossification centers of the body of sphenoid bone in the human fetus

The aim of the present study was to examine the growth dynamics of the two ossification centers of the body of sphenoid bone in the human fetus, based on their linear, planar and volumetric parameters. The examinations were carried out on 37 human fetuses of both sexes aged 18–30 weeks of gestation, which had been preserved in 10% neutral formalin solution. Using CT, digital image analysis software, 3D reconstruction and statistical methods, we evaluated the size of the presphenoid and postsphenoid ossification centers. The presphenoid ossification center grew proportionately in sagittal diameter, projection surface area and volume, and logarithmically in transverse diameter. The postsphenoid ossification center increased logarithmically in sagittal diameter, transverse diameter and projection surface area, while its volumetric growth followed proportionately. The numerical findings of the presphenoid and postsphenoid ossification centers may be considered age-specific reference values of potential relevance in monitoring the normal fetal growth and screening for congenital disorders in the fetus. The obtained results may contribute to a better understanding of the growing fetal skeleton, bringing new numerical information regarding its diagnosis and development.

Morphological features of the pons in human fetuses 14-15 weeks of intrauterine development

Knowledge of human embryonic development is essential to improve our understanding of human fetal anatomy and for better understanding the etiology of congenital malformations. Currently, the structures of the posterior cranial fossa and the brain stem are of great interest to researchers, because of a large number of nuclei are located in the pons area, which play an important role in ensuring vital functions. The aim of the scientific work is to establish the morphological features of the pons of human fetuses at 14-15 weeks of gestation, the size and area of the nuclei of cranial nerves and neurons which are located in the area of the pons. Anatomical and histological research was carried out on 6 human fetuses aged 14-15 weeks of gestation. The cadaver material for research was obtained as a result of late abortions which were conducted according to medical indications in Vinnytsia maternity hospitals. Preparations were fixed in a 10 % solution of neutral formalin, stained with hematoxylin-eosin, toluidine blue modified by Nissl. Computer histometry (Toup View) was used for morphometric research. Statistical digital data were processed on a personal computer using Microsoft Excel 2016 and “Statistica 6.1” software. We established that in human fetuses at 14-15 weeks of intrauterine development, the nuclei of the trigeminal, abductor and vestibulocochlear nerves were detected. The nucleus of the facial nerve on 14-15 weeks of gestation is represented by single polygonal nerve cells with eosinophilic cytoplasm. The nucleus of the abductor nerve had the largest area, the cochlear nucleus of the vestibulocochlear nerve had the smallest area. The largest area of neurons was in the nuclei of the VIII pair of cranial nerve, the smallest – in the nucleus of the abductor and facial nerves. At 14-15 weeks of intrauterine development of human fetuses, the area of the cell nucleus could be determined only in the nuclei of the vestibulocochlear nerve, while the cell nuclei of the trigeminal, facial, and abductor nerves were represented by nerve cells of a spherical shape with a nucleus in which a basophilic nucleolus and a homogeneous eosinophilic cytoplasm were noted. Thus, in human fetuses of 14-15 weeks of intrauterine development, differences in the sizes and areas of the nuclei of cranial nerves and neurons that form nuclei in the pons area were found.

Features of the microstructural organization of the small intestine in weaning piglets

The material for the study was fragments of the wall of the duodenum, jejunum and ileum obtained from five piglets at the age of 41 days. The selected tissue samples were fixed during the day in a 10.0% solution of neutral formalin. Their further processing in order to obtain histological sections was carried out according to a generally accepted method. The obtained histological sections were stained with hematoxylin and eosin and according to Van Gieson. All morphometric data obtained were subjected to statistical processing.It was found that in the studied animals, the small intestine is arranged in accordance with the general plan of structure peculiar to mammals. Its wall is formed by the mucous membrane, submucosal base, as well as muscle and serous mem-branes. The epithelial layer of the mucous membrane of the small intestine is represented by a single-layer prismatic epithelium, which is a highly dynamic structure that quickly reacts to changing loads due to the high rate of cell renewal. The bulk of the epithelial layer is represented by edged enterocytes (about 90.0% of the entire cell population). In addition to them, goblet-shaped, endocrine, Panethovian, proliferating and stem cells take part in the formation of the epithelial lining of the small intestine.

HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL FEATURES OF DIFFERENTIATED TROPHOBLAST IN CHORIONIC VILLI OF THE PLACENTA IN PRETERM LABOR

The importance of studying the histochemical and immunohistochemical features of the diff erentiated trophoblast in chorionic villi of the placenta in preterm labor is determined by the threatening nature of this condition for both mother and fetus, as well as its high prevalence and serious consequences for the health of both. The understanding of the damage to the trophoblast of chorionic villi in the placenta can be expanded by histochemical and immunohistochemical methods, which allow the assessment of the concentration of specifi c marker molecules in one way or another.Objective. To determine certain histochemical and immunohistochemical characteristics of proteins in the trophoblast of theintermediate and terminal chorionic villi of the placenta in preterm labor.Material and Methods. The obtained material (30 placentas from preterm deliveries and 30 placentas from normalpregnancies) was fi xed for 20-24 hours in 10 % neutral formalin solution buff ered in Lilly’s phosphate buff er. After tissue removal, the placental tissue was dehydrated in an ascending ethanol series and embedded in paraffi n at a temperature of approximately 58 °C. Serial histologic sections were cut at 5.0 μm thickness using an MS-2 sliding microtome. After deparaffi nization, histological sections were stained with hematoxylin and eosin, histochemical methods for total protein with bromophenol blue according to Bonhéme, and immunohistochemical techniques according to the manufacturer’s protocols (Dako, Denmark). In particular, immunohistochemical reactions were performed with monoclonal antibodies against trophoblast hormone- placental lactogen and placental alkaline phosphatase. Visualization of primary antibodies was performed using the Dako polymer visualization system with diaminobenzidine as chromogen (resulting in brown staining of the sites of studied antigens). In addition to the descriptive method of histopathologic research, computer morphometry of digital microphotographs of histologic sections was performed using a Delta Optical Evolution 100 microscope and an Olympus SP550UZ digital camera. Digital copies of the images were processed using a legitimate copy of the ImageJ v1.52f computer program developed for histometric studies (National Institutes of Health, USA). Specifi cally, the evaluation of staining intensity (optical density) was performed on digital microphotographs using the method of computer microdensitometry. For this purpose, a microprobe method was used to obtain a computer brightness value in an 8-bit analysis system with 256 gray levels – from black (0) to white (255).The obtained values were then transformed into relative optical density values (r.OD) by logarithmic transformation (naturallogarithm method). The relative optical density value ranges from 0 (absolute transparency of the object) to 1 (absolute opacity of the object). The obtained digital data were processed using statistical analysis methods. A legitimate copy of the statistical analysis computer program PAST v4.14 was used, with a preliminary check for normal distribution using the Shapiro- Wilk test. Since, according to this test, the hypothesis of normal distribution was not rejected for the statistical samples studied (at p=0.05), parametric methods of statistical analysis were applied: calculation of the mean and its standard error, Student’s t-test (two-tailed, unpaired). In addition, the non-parametric Mann- Whitney test was used for the reliability of the conclusions. The research was conducted as part of the scientifi c research project «Preservation and Restoration of Reproductive Health in Women and Girls with Obstetric and Gynecological Pathology» at the Department of Obstetrics and Gynecology of Bukovinian State Medical University (state registration number 0121U110020, duration 2021-2025).Results. Microscopic examination of histological sections stained with hematoxylin and eosin did not reveal any diff erencesin the structure of chorionic villus trophoblast in the placenta of preterm labor compared to normal pregnancy. However,histochemical and immunohistochemical methods of investigation revealed a number of features in the trophoblast of chorionic villi, where fundamental events related to substance exchange occur – intermediate immature, intermediate mature, terminal villi, including terminal «specialized villi».Conclusions. According to the obtained histochemical and immunohistochemical data, in preterm labor, compared to normalpregnancy, no changes are observed in the trophoblast of intermediate immature villi, while in intermediate mature and terminal villi, there is a decrease in histochemical staining for total protein and immunohistochemical staining for specifi c trophoblast proteins – placental lactogen hormone and placental alkaline phosphatase enzyme.

The effect of the irrigation solutions on dentin organic components: Pilot study

INTRODUCTION. Tooth decay and its complications are prevalent dental issues among adults. Endodontic treatment is the standard of care for complicated caries, specifically pulpitis and periodontitis. Endodontic irrigants, which are placed in the root canal lumen for a specific duration, can affect the intracanal dentin structure (organic and inorganic substances) in various ways. The use of polyhexanide-based antiseptic products in endodontics is a promising idea to overcome microbial resistance and minimize microbial-related endodontic treatment failures.AIM. The aim of this study is to investigate the qualitative impact of standard irrigation solutions and a polyhexanide-based composition on the collagen matrix of dentin.MATERIALS AND METHODS. The study investigated the effect of nine solutions on the dentin of the tooth root. Twelve intact maxillary third molars extracted for orthodontic reasons were taken for the study. The teeth were fixed in a 10% neutral formalin solution for at least one day and were decalcinated. 5–10 μm thick sample sections were made and stained with picrosirius (Picrosirius Red, Biovitrum, Russia) before being examined under polarized light. The histological slice was analyzed using a Carl Zeiss (Germany) software and hardware complex based on ZEN v3.0 and light microscope Axioimager M.1. All microphotos were loaded into the Fiji distribution program of ImageJ for microscopy pixel classification.RESULTS. Upon analysis of the diagrams, it appears that there is a lower amount and density of stained dentin collagen array after exposure to sodium hypochlorite solutions of 3% and 1.5%, as well as polyhexanide.CONCLUSIONS. The data obtained from the histogram analyzer of samples stained with picrosirius suggests that polyhexanide may have a qualitative effect on the organic structure of hard tissues, particularly collagen.

MORPHOLOGICAL CHANGES OF RAT PERIODONTIUM IN SUPRAOCCLUSAL RELATIONS OF INDIVIDUAL TEETH IN THE AGE ASPECT

Although the issue of the occlusal load influence on the occurrence of pathological phenomena in the body has been discussed for decades, it is still a subject of debate. Pathogenic occlusion is defined as excessive occlusal forces (premature contacts and interference) that can cause injuries to the dentition, including teeth and periodontal tissues. Changes in occlusal dimensions, if not properly performed or treated, can lead to dysfunctions of the dentition, such as joint clicking, difficulty in chewing, occlusal discomfort, muscle fatigue, and pain.
 Aim: to determine the morphological changes of periodontium in rats of different age groups in the supraocclusal relationship of individual teeth in the age aspect.
 Materials and methods of the study. The study was conducted on 60 white laboratory rats aged 4 to 22 months and weighing 90 to 330 g. The rats were divided into control (30 animals) and experimental (30 animals) groups. Each group was divided into 3 subgroups (10 animals each) depending on the age of the rats: young, mature and senile. Young animals included rats aged 3 4 months with a weight of 90 100 g; mature animals aged 6-8 months with a weight of 150 170 g; senile rats 20 22 months and weighing 200 220 g. In the experimental group, the state of supraocclusion was modeled by increasing the height of the lower right second molars by placing 1 mm high filling. For the placement of the filling, 37% orthophosphorus etching gel for enamel and dentin (Latus, Ukraine), microapplicators (Latus, Ukraine), the fifth generation Prime&Bond NT adhesive system (Dentsply Sirona, Germany), the universal microhybrid light-curing composite material Latelux (Latus, Ukraine) and the photopolymerizer LED.D (WOODPECKER, China) were used according to the manufacturer's instructions. The experimental animals were withdrawn from the experiment on the 15th day by decapitation under general anesthesia with sodium thiopental. The lower jaws were skeletonized, and dental blocks were cut out in the area of the molars. For histological examination, the dental blocks were fixed in a 10% solution of neutral formalin for two days, dehydrated in alcohols of increasing concentration and embedded in paraffin. Then, sections were made from the obtained preparations on a Shandon Finesse 325 rotary microtome with a section thickness of 4-6 μm. The histological sections were fixed on degreased slides and dried. Van Gieson staining was performed. The width of the periodontal gap was determined using the ZEISS ZEN 3.7 program by Carl Zeiss Microscopy.
 Results of the study. Histometric analysis has showed significant morphological changes in the experimental group of all age categories. In young rats, the fibrinoid edema phenomena of the periodontal ligament collagen fibers were observed. In mature rats, there was a picture of mucoid edema of collagen fibers of the periodontal ligament. In senile rats, fibrinoid necrosis of collagen fibers of the periodontal ligament were discovered, an inflammatory infiltrate, macrophages and neutrophils, with a predominance of neutrophilic inflammatory microenvironment, and initial hyaline tissue changes were found. In all age groups, there was cement resorption on the lateral surfaces of the tooth root, lacunar resorption of alveolar bone tissue and the presence of a large number of osteoclasts. In senile rats, hypercementosis was determined in the apical part of the root. In the control group of all age categories, neither resorption nor disorganization of connective tissue, nor hypercementosis was detected.
 Conclusions. The study shows that traumatic occlusion leads to resorption of root and alveolar cementum in all age groups. In young rats, traumatic occlusion causes fibrinoid edema of collagen fibers of the periodontal ligament. In mature rats, traumatic occlusion causes mucoid edema of collagen fibers of the periodontal ligament. In senile rats, traumatic occlusion causes fibrinoid necrosis of collagen fibers of the periodontal ligament, initial hyaline tissue changes and hypercementosis.

Histomorphology of the ovaries of rabbits does during ovulation induced by the combined use of gonadotropins

Hormonal stimulation of folliculogenesis is a general method of increasing reproductive capacity in rabbits; however, studying the safety of the developed protocols remains an urgent scientific task. The use of hormonal agents is due to the physiological characteristics of rabbits, primarily coitus-dependent mechanisms of ovulation. Applying gonadotropin-releasing analogs depends on mature follicles ready for ovulation in the ovary. It, therefore, has limited effectiveness for rabbits of the “zero” cycle that have not yet given birth. Therefore, such females are recommended to use medications based on gonadotropins obtained from the serum of foal mares (eCG) or human chorionic gonadotropin (hCG). There is limited data on the combined use of gonadotropins, especially the long-term administration in different doses. Thus, the research aimed to conduct a morphological evaluation of rabbit's ovaries with combined serum and chorionic gonadotropins to stimulate folliculogenesis before artificial insemination to assess the safety of the proposed dosages and long-term administration. Stimulation of ovulation in rabbits of experimental groups was caused by the combined use of serum and chorionic gonadotropins (eCG – 400 IU; hCG – 200 IU) for experimental groups 1 and 2 at a dose of 40 IU and 24 IU, respectively. Females of the control group were induced to ovulate by subcutaneous injection of 0.2 ml of gonadotropin-releasing hormone analog. Ovaries were collected on the seventh day of gestation after the euthanasia of the rabbit with prior premedication. After dissection and macroscopic examination, the ovaries were fixed in a 10% aqueous solution of neutral formalin. Histological slides were made according to the general method by pouring in paraffin and subsequent staining with hematoxylin and eosin. The research has established that repeated (during five reproductive cycles) combined use of serum and chorionic gonadotropins has dose-dependent effectiveness and can cause dysfunction of ovaries and blood circulation. Pathological changes may acquire a systemic character with the manifestation of critical conditions for the health and life of rabbits. The stimulating effect of the combined use of eCG and hCG was detected at a dose of 40 IU, and the inhibition of folliculogenesis was observed at 24 IU. Since the premises for the hyperandrogenism were found microscopically, it was essential to investigate the level of androgens in the blood of rabbits after using a dose of 24 IU. At the same time, both doses of gonadotropins affect the reactivity of the endothelium, which is manifested by the presence of intravascular vesicles and changes in the rheological properties of blood.

Morphological characteristics of the kidneys of mature dogs (Canis Lupus Familiaris L., 1758)

Each biological species of animals is unique in its structure, which is due to the uniqueness of ecological habitats and the peculiarities of adaptation of homeostasis mechanisms to the influence of environmental factors. Subtle differences in the structural and functional components of organs found in different species of animals create a clear architecture for understanding the depth of biological processes. Research of species, breed and age macro- and microanatomical structures and their morphometric indicators of organs and systems should be considered as urgent tasks of modern morphology. The new data on the morphological indicators of organs discovered at the macro- and micro-levels make a certain contribution to the age-related, breed-related, comparative, experimental and clinical morphology of mammals. Urinary organs in the body of mammals play one of the leading roles in maintaining the physiological and biochemical status of the organism and homeostasis as a whole, which largely depends directly on the morphological and functional state of these organs. Increasingly, there is a need for comprehensive scientific studies of various departments of the urinary organs. The scientific article uses material that is a fragment of the planned research work of the department of normal and pathological morphology, hygiene and expertise “Development, morphology and histochemistry of animal organs in normal and pathological conditions”, state registration number 0120U100796. The article contains the results of the morphometric characteristics of the kidneys of sexually mature dogs – Canis lupus familiaris L., 1758. During the implementation of this work, complex research methods were applied: anatomical, histological, morphometric and statistical. For histological examination, a fragment of kidneys was selected, they were fixed in 10 % neutral formalin solution, Carnois and Buena fluid. In the future, dehydration was carried out in alcohols of increasing concentration and the procedure for sealing the organ was carried out according to generally accepted methods. Using a sled microtome, histological sections with a thickness of 7–10 μm were obtained. Processing of the results of histological sections was examined under a “Micros” light microscope with a digital camera at a magnification of × 100–400. The morphological study of the structural organs of the urinary system was studied on histological sections by the method of light microscopy in accordance with the requirements of the international principles of the “European Convention for the Protection of Vertebrate Animals Used in Experiments and Other Scientific Purposes” (Strasbourg, 1986), by the decision of the First National Congress on Bioethics (M. Kyiv, 2001), Law of Ukraine No. 692 “On the Protection of Animals from Cruel Treatment” (3447-IV) dated February 21, 2006. According to the histological structure, the kidneys of dogs are represented by renal corpuscles and renal tubules, which are made of epithelial cells. Organometric studies indicate that the absolute mass of kidneys in sexually mature dogs is 119.8 ± 6.48 g, relative – 0.74 ± 0.04 %. Morphometric studies of the kidney parenchyma in dogs have established that the average area of renal corpuscles is 4.89 ± 0.24 thousand μm², the area of the vascular glomerulus is 3.79 ± 0.18 thousand μm², the area of the Shumlyansky-Bowman capsule lumen is 1.12 ± 0.27 thousand μm². The average volume of renal bodies of the cortical substance is 1265.69 ± 239.34 thousand μm³. The number of renal corpuscles per conventional unit of area (5 mm²) is 20.8 ± 0.6 units. The aim of the work was to investigate the peculiarities of the morphogistological structure of the kidneys of sexually mature dogs. The research results will expand information about the morphological structure, and will serve for the development of issues of physiology, pathology and treatment of diseases of the urinary organs.

CHANGES IN THE WIDTH OF THE PERIODONTAL GAP OF RATS WITH SUPRAOCCLUSAL RELATIONSHIPS OF INDIVIDUAL TEETH IN THE AGE ASPECT

Introduction. The dentoalveolar apparatus is a complex of complicated interaction of the jaws, masticatory muscles, teeth, and temporomandibular joints. This connection is carried out by the trigeminal nerve system with sensory and motor nuclei closely connected to the cortical and subcortical centers of the brain. Occlusal trauma is a lesion that develops in the periodontium as a result of forces that exceed the periodontal potential for functional adaptation. Occlusal trauma can be classified as primary or secondary. The first one occurs in the pulp and periodontal ligament when the force of the action on the dental structures is greater than usual, under optimal periodontal support conditions. It includes orthodontic movements, "high" restorations, and parafunctional habits such as bruxism. Secondary occlusal trauma occurs when the teeth with previous supporting tissue changes are subjected to the action of normal and excessive forces with supporting tissue changes caused by periodontal disease or previous trauma. The available evidence has demonstrated that bacterial plaque bioactive substances lead to local inflammatory responses in periodontal tissues which contribute to bone tissue resorption through osteoclasts in periodontitis. Excessive functional stress can initiate inflammatory changes in the periodontium and thus, increase destructive bacterial processes. Traumatic occlusion can expand the space of the periodontal ligament and in this way stimulate apical bacterial migration and proliferation. However, few attempts have been made to investigate the molecular mechanisms that trigger bone tissue resorption in traumatic occlusion. The analysis of publications on the studies of morphological changes in periodontal tissues in individual teeth supraocclusion in the age aspect shows the contradictory data. The aim: to determine the morphometric changes of the periodontal gap as a component of the periodontal complex in rats of different age groups in the supraocclusal relationship of individual teeth in the age aspect. Materials and methods of the study. The scientific experiment was conducted on 60 white laboratory rats aged 4 to 22 months and weighing 90 to 330 g. The rats were divided into control (30 animals) and experimental (30 animals) groups. Each group was divided into 3 subgroups (10 animals each) depending on the age of the rats: young, mature and senile. Young animals included rats aged 3–4 months with a weight of 90–100 g; mature animals - aged 6–8 months with a weight of 150–170 g; senile rats – 20–22 months and weighing 200–220 g. The method of modelling the state of supraocclusal relations of individual teeth is described in the previous publication. For histological examination of dental blocks, the following was performed. Fixation in a 10% solution of neutral formalin for two days, dehydration in alcohols of increasing concentration and embedding in paraffin. After preparation, sections were made on a Shandon Finesse 325 rotary microtome with a section thickness of 4-6 µm. The histological sections were fixed on pre-degreased slides and dried. Van Gieson staining was performed. The width of the periodontal gap was determined using the ZEISS ZEN 3.7 program by Carl Zeiss Microscopy. For statistical data processing, the AtteStat v.12.5 software was used to determine the mean value and its standard error (M±m). The probability of a statistically significant difference between the data obtained was assessed using the Student's parametric test (at a significance level of p ≤ 0.05). All samples were tested for normality using the Gupta test. Results of the study. Statistical differences were observed in rats of all groups, as well as between the indicators of all rats of the control and experimental groups (≤0.05). The width of the periodontal gap increased by 34.1% in young rats, by 25.5% in mature rats, and by 21.1% in senile rats. Conclusions The presence of supraocclusal relationships of individual teeth causes an increase in the periodontal gap in rats of all age groups. The greatest expansion of the periodontal gap has occurred in young rats. The width of the periodontal gap in rats decreases with age.

Screening of anti-inflammatory activity of 4.5-dihydroisoxazol-5-carboxamide (PAR-2 inhibitors) based on formaldehyde oedema model among white lab rats


 Introduction: Rheumatoid arthritis (RА) is an immune-inflammatory disease intrinsic to up to 1.0% of the world’s population. Standard drugs for RA therapy are basic medications, glucocorticoids and non-steroid anti-inflammatory drugs, which often only ease or hinder the course of disease, not curing the patient completely. Also, on the average 20-50 % of patients are unresponsive to treatment, allergic to the prescribed drugs or find them ineffective. That is why medications with a different mechanism of action are being widely researched, some of them being antagonists of proteinase-activated receptors (PAR-2).
 
 
 Materials and Methods: The inflammatory process was reproduced by injecting a 2% solution of neutral formalin in a volume of 0.1 mL under the aponeurosis of the posterior left foot. A total of 84 white rats were used in the experiment. Diclofenac sodium was administered as a reference drug.
 
 
 Results and Discussion: An experiment on researching the impact of 5 samples of derivatives of 4.5-dihydroisoxazol-5-carboxamide on formalin oedema development among rats showed that the compound R001, compared with other substances studied, differs in the late onset of the therapeutic effect when ingested; the restoration of the foot volume to the initial level occurs only after the introduction of R005, R004 and R002. R005 to a greater extent than other compounds prevents the development of edema and has greater therapeutic efficacy than diclofenac sodium both with intragastric administration and subcutaneous injection.
 
 
 Conclusion: All five compounds in question showed anti-inflammatory activity, with the spectrum not so unequivocal both in qualitative and quantitative values.

RATS’ THYROID GLAND HISTOLOGICAL AND ULTRASTRUCTURAL CHANGES THROUGHOUT THE EXPERIMENTAL THERMAL INJURY DYNAMICS ON THE BACKGROUND OF HAES-LX 5 % COLLOID-HYPEROSMOLAR SOLUTION INJECTION

The urgency of the burn injury problem is determined by frequent thermal injuries of patients of different ages, the complexity and duration of treatment, long-term disability and relatively high mortality. Depending on the area and depth of the lesion, a burn wound causes multiple and long-lasting disturbances in homeostasis, which cause organs and systems dysfunction. We are interested in the intrathyroid changes that occur during thermal burns, as thyroid gland plays one of the leading roles in the endocrine regulation of organisms’ majority functions. When researching the issue of pharmacological correction of thyroid gland damage after a burn injury, we proceed from the classical fundamental concept regarding its pathogenetic validity. Taking into account the known pathogenetic mechanisms of burn disease with consecutive (and sometimes simultaneous) hypoproteinemia and haemoconcentration manifestations, intoxication formation, inflammatory and autoimmune reaction, we came to a conclusion regarding the reasonability of colloid solutions efficacy testing to attempt the pharmacocorrection in case of thyroid gland both structure and function burning. The purpose of the work is to establish histological and ultrastructural changes in the thyroid gland of experimental rats that were injected with colloidal hyperosmolar solution HAES-LX 5 % in the dynamics of skin thermal damage. Experimental studies were conducted on 90 white male rats. Skin thermal burns were simulated using four copper plates application to previously depilated lateral surfaces of the rats’ body for 10 s. Rats were injected with colloidal hyperosmolar solution HAES-LX 5 % into the vena cava inferior during the first 7 days of the post-burn period. Thyroid gland pieces were fixed in a 10 % neutral formalin solution, dehydrated in alcohols of increasing concentration and embedded in paraffin blocks. The prepared sections of 5-6 μm thickness were stained with hematoxylin-eosin. For electron microscopic studies, pieces of the thyroid gland were taken, fixed in a 2.5% glutaraldehyde solution, and postfixed with a 1% osmium tetroxide solution in a phosphate buffer. Semi-thin sections were stained with methylene blue. Ultrathin sections were contrasted with uranyl acetate, lead citrate according to the Reynolds method and studied in a PEM-125K electron microscope. Colloidal hyperosmolar solution HAES-LX 5 % administration within 7 days of the post-burn period to correct the thermal injury effects has an expressed positive effect on burned animals thyroid gland histo- and ultrastructure. A significant improvement of the structural state of the stromal and parenchymal components of the organ and their relative normalization in the late period under the influence of the applied solution was established in the dynamics of the experiment. The colloidal hyperosmolar solution HAES-LX 5 % positive effects were expressed by cellular walls of the vessels and follicles dystrophic and destructive changes reduction the structural components of the organ restoration during the entire period of the study up to the 30th day of the trial. The first signs of the intraglandular environment recovery after colloidal hyperosmolar solution HAES-LX 5 % use were proved to start registered from the 7th day of the post-burn period and were maximally expressed from the 21st day until the end of the experiment. The authors are sure that colloidal hyperosmolar HAES-LX 5 % solution protective action possible mechanism is the generalized catabolic reaction inhibition and the membrane-protective effect development. A complex of colloidal hyperosmolar solution HAES-LX 5 % protective, adaptive, adaptive, compensatory and regenerative effects were realized throughout the 30 days of the post-burn period, which efficacy exceeds the thyroid gland parenchyma and surrounding tissues destructive, decompensatory and necrotic changes. The authors consider the use of colloidal hyperosmolar solution HAES-LX 5 % to be one of the burn treatment regimen components as a restorative therapy drug and secondary cytoprotection aimed at the vascular wall and tissue defects integrity restoring.

Structural organization of the pontine nuclei in human fetus 39-40 weeks of intrauterine development

Annotation. The study of the embryonic development of the human brain is important for understanding the early mechanisms of CNS development and assessing the influence of intrauterine factors on the formation of brain structures. Basic knowledge about the development of the fetal pons is important for any prenatal assessment in cases of suspected abnormalities of brain development. The aim of our study is to define the macrometric parameters of the human fetal pons at the stage of 39-40 weeks of fetal development, the size and area of the cranial nerve nuclei, which are located in the pons, as well as a morphometric study of neurons of the nuclei of cranial nerves. Anatomical and histological research was carried out on 5 human fetuses with a gestation period of 39-40 weeks. The material for the study was obtained from the Vinnytsia Regional Pathology Bureau, stillborns who died from causes not related to diseases of the central nervous system. Brain preparations were fixed with 10% neutral formalin solution. Digital data were statistically processed on a personal computer using Microsoft Excel 2016 and the software “Statistica 6.1” (license number BXXR901E246122FA). Computer histometry (Toup View) was used during the morphometric study. We found that in human fetuses at 39-40 weeks of gestation, the principal nucleus of the trigeminal nerve had the largest area. The dorsal cochlear nucleus of the vestibulocochlear nerve had the smallest area. The largest area of neurons was in the nucleus of the facial nerve. The largest diameter and area of cell nuclei has the principal nucleus of the trigeminal nerve, while the diameter and area of cells of nuclei of the facial, abductor and vestibulocochlear nerves have almost the same size.

Quantitative anatomy of the primary ossification center of the squamous part of temporal bone in the human fetus

Detailed numerical data about the development of primary ossification centers in human fetuses may influence both better evaluation and early detection of skeletal dysplasias, which are associated with delayed development and mineralization of ossification centers. To the best of our knowledge, this is the first report in the medical literature to morphometrically analyze the primary ossification center of the squamous part of temporal bone in human fetuses based on computed tomography imaging. The present study offers a precise quantitative foundation for ossification of the squamous part of temporal bone that may contribute to enhanced prenatal care and improved outcomes for fetuses with inherited cranial defects and skeletodysplasias. The examinations were carried out on 37 human fetuses of both sexes (16 males and 21 females) aged 18–30 weeks of gestation, which had been preserved in 10% neutral formalin solution. Using CT, digital image analysis software, 3D reconstruction and statistical methods, the size of the primary ossification center of the squamous part of temporal bone was evaluated. With neither sex nor laterality differences, the best-fit growth patterns for the primary ossification center of the squamous part of temporal bone was modelled by the linear function: y = −0.7270 + 0.7682 × age ± 1.256 for its vertical diameter, and the four-degree polynomial functions: y = 5.434 + 0.000019 × (age)4 ± 1.617 for its sagittal diameter, y = −4.086 + 0.00029 × (age)4 ± 2.230 for its projection surface area and y = −25.213 + 0.0004 × (age)4 ± 3.563 for its volume. The CT-based numerical data and growth patterns of the primary ossification center of the squamous part of temporal bone may serve as age-specific normative intervals of relevance for gynecologists, obstetricians, pediatricians and radiologists during screening ultrasound scans of fetuses. Our findings for the growing primary ossification center of the squamous part of temporal bone may be conducive in daily clinical practice, while ultrasonically monitoring normal fetal growth and screening for inherited cranial faults and skeletodysplasias.

PATHOMORPHOLOGICAL CHANGES IN THE ORGANS OF CATS CAUSED BY ALIMENTARY LYMPHOMA. CLINICAL CASES

The article presents the results of clinical, ultrasonographic, pathomorphological and immunohistochemical studies of two random cases of alimentary lymphoma in 9- and 12-year-old domestic cats. Clinical signs of the disease were similar and characterized by anorexia, cachexia, vomiting and diarrhea that appeared in different periods of the disease. The mucous membranes of the oral cavity and eyes were icteric, the abdomen was visually enlarged, palpation revealed nodular thickenings in the abdominal cavity. During the ultrasound diagnostics, diffuse multiple non-encapsulated anechoic formations with uneven edges and growth of tumorous tissue into the intestinal wall were visualized. In another cat, there was an encapsulated formation of a hypoechoic fine-grained structure with well-defined vascularization.
 From the moment of the preliminary diagnosis until the very death, the cats were kept in the clinic under supervision. After the death, an autopsy was performed with the selection of material for histopathological examination. Organs’ fragments were fixed in 10% aqueous solution of neutral formalin and Carnois fluid. Histological preparations were made according to the generally accepted methods. Immunophenotyping of lymphoma type was performed using CD3 (T-cell) and CD79a (B-cell) markers. During the pathological autopsy, an increase in mesenteric lymph nodes, their compaction, and focal thickening of the small intestine wall were revealed. The histological examination showed that jejunum was the most infiltrated intestinal segment, in which massive, diffusely placed monomorphic lymphoblastic cells were localized in the mucous membrane lamina, submucosa and muscle layers. The submucosal layer vessels were sharply dilated, filled with erythrocytes; the muscles of the inner circular layer were swollen. The outer (longitudinal) muscle layer of the jejunum thickened significantly and was intensively infiltrated by lymphoblastic cells.
 Typical diffuse lymphoblastic proliferations were also detected in the cortical substance of lymph nodes and lymphoid nodules. Stasis and significant expansion of the marginal sinus vessels were visualized too. In addition, moderate mononuclear lymphoblastic infiltrates were detected in the pancreas parenchyma with their perivascular location. Immunohistochemically, a significant positive expression of CD3 cells was detected both in the jejunum mucous membrane and the marginal sinus area, around the germinal centers of the lymph nodes, which indicated a T-cell type of lymphoma.

Microarchitectonics and histomorphometric parameters of bone tissue during remodeling

The article shows and statistically substantiates thepeculiarities of remodeling and structure of cortical andmedullary bone tissue in two age groups (160- and 225days) of laying hens before and during egg-laying. Theaim of the study was to analyze the structural organization of bone tissue in different states of performance.The material for the study was the ulnae of the forearmof laying hens of two experimental groups. Histological studies: fixed in a 10% solution of neutral formalin,decalcified in Wilenson's fluid, dehydrated in alcoholsof increasing concentration, embedded in paraffin, prepared transverse sections from the middle third of thediaphysis, stained by Frankel, Hart, Malory, hematoxylin and eosin. Microscopy and morphometric studies were performed using an Axiolar plus microscope(Carl Zeiss), photographs were taken and measuredusing a Sigeta MDC-200 video camera (eyepiece x10,lenses x25/0.50, x40/0.65, 100/0.25) integrated with anAxiolar plus personal computer (Carl Zeiss). Statisticalprocessing of digital material was performed by Student's t-test. Changes in the structure of cortical bonetissue during the study period are associated with physiological remodeling. This is evidenced by the presenceof cementing lines of individual osteons, semi-osteonsand Haversian canals with an enlarged diameter. Theabsence or insignificant development of the insertionsystem of the forming bone plates indicates a low rateof physiological remodeling of the compact bone substance. The external general system of plates does notchange the structure. The internal general system ofbone plates is unevenly developed. From 160 to 225days of age, the development and accumulation ofmedullary bone tissue occurs, which is confirmed bya significant increase in the area of trabeculae by 2.8times (P3 = 0.999), a complication of their structureand a decrease in the area of intertrabecular spaces by1.7 times (P2 = 0.99). A significantly higher 3.4-fold(P1 = 0.95) volume of osteoblast nuclei at 160 days indicates their active functional activity - the formationof trabeculae of medullary bone tissue. During the period of 160 days, special attention should be paid to thebalance of the diet of laying hens to ensure the formation of a sufficient amount of high-quality medullarybone tissue. Key words: lamellar bone tissue, cortical, medullary, spongy and compact substance, restructuring,histomorphometry, laying hens.

Histomorphology of the vesicular and prostate glands of newborn piglets

The economic efficiency of animal husbandry depends on the productivity of the herd, which is usually assessed by analyzing the reproductive performance of females. However, similar indicators of males should be equally taken into account. The accessory sex glands of males are involved in the production of seminal plasma, thereby playing an important role in the reproductive process. The aim of the study is to give a histomorphological characteristic of the accessory sex glands and to determine their functional state in newborn piglets. The object of the study was the accessory sex glands of newborn piglets at the age of 1 day. The vesicular and prostate glands were selected for histological examination. The material was fixed in a 10% solution of neutral formalin for 24 hours, after which it was poured into paraffin according to the generally accepted method. Then sections with a thickness of 3- 5 microns were made, which were stained with hematoxylin and eosin. Some of the sections were painted with Masson Trichrome, alcian blue. It was found that in newborn piglets, the vesicular gland is located on the dorsal surface of the bladder, at the beginning of the pelvic part of the urethra, the prostate gland is located caudally from the vesicular gland, dorsally from the genitourinary canal. The study established the main histomorphological features of the vesicular and prostate glands in newborn piglets. The parenchyma of the glands is in the stage of functional immaturity, most of the organ is occupied by the connective tissue stroma, among the elements of which there is a large number of fibroblastic cells. At the same time, secretory activity of some end sections of the vesicular gland is noted. The data obtained are the starting point for further studies of the functional activity of the cells of the accessory sex glands in boars.

Structural-functional changes in the proventriculus of poultry infected with transmissive viral proventriculitis

Transmissive viral proventriculitis (TVP) is a new avian disease, the etiology and pathogenesis and diagnostics of which remain unidentified, being therefore a subject of an active ongoing discussion among researchers and veterinary doctors. Cases of the disease have been reported and registered in many countries of the world. On poultry farms of Ukraine, this disease started to be diagnosed in 2019, according to the characteristic pathognomic changes. The article presents the results of pathoanatomical, histological and electronic-microscopical studies of the glandular part of the stomach of 62 ill broiler chickens aged 24-28 days and 7 clinical healthy broiler chickens of the same age. The samples of the proventriculus from the infected and clinically healthy chickens were fixated in 10% aqueous solution of neutral formalin and in 1.5% solution of glutaraldehyde in 0.2 molar cacodylate buffer (рН 7.2). Pathoanatomic necropsy of the infected broiler chickens revealed overall anemia, decrease in the volume of the spleen, thymus and fecal sac. In the glandular part of the stomach, the wall was slightly thickened, non-homogenously stained on the section, the mucous membrane being swollen, covered by dense whitish mucous. According to the histological study, the proventriculus of the broiler chickens had lymphocytic necrotizing proventriculitis. In the tubular-alveolar structures of the mucous membrane, there prevailed alteration of the secretory epithelium, with its transformation into ductal epithelium, combined with lymphocytic infiltration of the interstitium. Intensive necrosis of most glandular (oxynticopeptic) cells was accompanied by their exfoliation from the basal membrane of the secretory parts, with desquamated cells accumulating in enlarged lumens of alveoli and excretory ducts. Further, necrotized oxynticopeptic cells in the tubular-alveolar glands were actively replaced by cylindrical ductal cells, and also their hyperplasia and hypertrophy occurred. Diffusive and multifocal interalveolar round-cell infiltrates were mostly presented by lymphocytes, among which there occurred plasmocytes, macrophages, and heterophils. Ultrastructural studies of secretory cells revealed presence of pathogen virions in nucleus and cytoplasm. Based on the morphological changes in the birds’ proventriculus, we diagnosed transmissive viral proventriculitis, which had not been recorded in Ukraine before.

Morphological changes in the liver of rats after administration of chlorpromazine, depending on the dose and duration of administration

Chlorpromazine (CPZ) remains a widely used drug in psychiatric practice today. The drug has a hepatotoxic effect, but the possible mechanisms of this side effect have not yet been fully elucidated. The aim of the study was to determine morphological changes in rat liver tissue under chronic toxic effects of chlorpromazine, depending on the dose and duration of its administration. The study was conducted on 60 sexually mature male rats. CPP was administered intragastrically at different doses (3.5, 7.0, 14.0 and 21.0 mg/kg) for 30 and 60 days. The material was fixed in a 10 % solution of neutral formalin (pH 7.2-7.4) for 24-48 hours, then passed through alcohols of increasing concentration and embedded in paraffin. Serial sections (6-7 µm thick) were prepared from the paraffin blocks and stained with hematoxylin-eosin and picrofuchsin by Van Gieson to determine the degree of fibrotic changes in liver tissue, as well as with Giemsa III to detect fatty degeneration of hepatocytes. The microscopic structure of the hepatic parenchyma was studied using an OLIMPUS BX41 light microscope at 100, 200 and 400x magnification. Morphometric parameters of structural changes were determined using an ocular grid and Image Tulsa 3.6 software. The data were statistically processed by descriptive statistics using the Microsoft Office Excel 2010 spreadsheet processor. When CPZ was administered in different doses and duration, pathological changes of varying severity developed in the liver tissue of rats. In the liver tissue, signs of intracellular and intra-tubular cholestasis are found mainly in the central lobes, accompanied by focal desquamation and proliferation of the biliary epithelium, formation of small-focal, less frequently zonal necrosis of hepatocytes, inflammatory infiltration of portal tracts with its spread to the interlobular stroma and parenchyma. Mitotically active binucleated hepatocytes are the key to the reparative process. Periductal fibrosis develops in the portal sections, marginal proliferation of the bile ducts, hepatocytes with signs of granular and/or fatty dystrophy are noted. In the central veins and vessels of the portal areas, moderate initial sclerotic changes were found, signs of their capillarisation in sinusoids, and the endothelium of the vessels had focal destructive changes. In all portal zones, proliferation of bile ducts and formation of bile pseudo-ducts were observed. Thus, the analysis of the morphometric study data showed that within 60 days of CPZ administration there is a significant increase in the relative volume of connective tissue and stromal-parenchymal index due to a significant decrease in the volume of hepatocytes.