Ligature induced periodontitis causes atherosclerosis in rat descending aorta: an experimental study

Introduction. In recent years a link between periodontitis and cardiovascular diseases has been proposed. Oxidative stress has been found to be involved in their pathophysiology. Objectives. The aim of this study was to demonstrate that a wire ligature placed around the first mandibular molars of the rat causes inflammation in the periodontium and lipid deposits in the aorta, via oxidative stress. Methods. Twenty male Wistar rats were divided randomly into two groups of 10: Control group and Periodontitis group. An orthodontic wire ligature was placed around the cervix of the first mandibular molar and was kept in place for 4 weeks. Clinical attachment loss was determined to assess the gingival recession. Aorta was harvested after 4 weeks and the malondialdehyde (MDA) levels in the aorta tissue homogenate were determined. Lipid deposits were evaluated using Oil Red staining and immunofluorescence under confocal microscopy. Total lipid deposits in the aorta were assessed using a computer morphometry system. Results. Significant gingival recession was observed in the Periodontitis group in comparison with the Control group. Increased MDA levels were present in the aorta in the ligatured rats in comparison with the Control group. Significant lipid deposits were observed in the aorta of the Periodontitis group as opposed to the Control group. Conclusions. The affixing of an orthodontic wire ligature produces gingival recession, increased oxidative stress and lipid deposits in the rat descending aorta.

DIAGNOSTIC ROLE OF MORPHOMETRIC ASSESSMENT OF TUMORASSOCIATED STROMA COMPONENTS OF SEROUS OVARIAN CARCINOMAS IN THE PROGNOSIS OF TUMOR PROGRESSION

The method of computer morphometry of stroma components and evaluation of stromal-parenchymal relationships in the primary tumor node of serous ovarian cancer was used. A high potential for malignancy of tumor tissue, characterized by the resumption of tumor growth after radical treatment, is observed with the predominance of the epithelial component over the stroma, with a small number of necrosis in the tumor node, low density of microcirculatory vessels in the tumor, and the predominance of fi broblastic elements in the cellular microenvironment of the tumorassociated stroma.

AGE-RELATED CHANGES IN THICKNESS OF CORTEX AND ITS LAYERS IN THE PARAMEDIAN LOBULE OF THE CEREBELLUM IN CHILDREN

Цель - изучение возрастных изменений толщины коры и ее слоев в парамедианной дольке мозжечка у детей. Материал и методы. Работа выполнена на постмортальном материале (62 мозжечка), полученном от детей в возрасте от рождения до 12 лет, умерших в результате травм без повреждений головного мозга. С помощью компьютерной морфометрии на окрашенных методом Ниссля фронтальных гистологических срезах коры, взятой билатерально в области парамедианной (тонкой) дольки (HVIIB) на вершине листков мозжечка, измеряли толщину коры, а также толщину ее молекулярного и зернистого слоев. Анализ количественных данных проводили в годовых интервалах. Результаты. В парамедианной дольке мозжечка увеличение толщины коры происходит в четыре этапа: в правом полушарии - от рождения к 1, 3, 5 и 9 годам, в левом полушарии - к 1, 5, 7 и 9 годам. Левосторонняя асимметрия толщины коры мозжечка отмечается у детей 1 и 2 лет, толщины молекулярного слоя - у детей 3 лет жизни. Правосторонняя асимметрия характерна для толщины зернистого слоя у детей 3 лет и поперечника коры, в целом, у детей 6 лет. Толщина коры и слоев в области парамедианной дольки мозжечка по среднегрупповым показателям достигает уровня взрослых людей к 9 годам. Выводы. Толщина коры мозжечка и ее слоев в области дольки H VII B увеличивается гетерохронно и гетеродинамически в правом и левом полушариях мозжечка у детей на первом году жизни, а также в периоды раннего, первого и второго детства. Уменьшения поперечника коры и слоев в парамедианной дольке мозжечка у детей от рождения до 12 лет не обнаружено. Objective - to study the age-related changes in the thickness of the cortex and its layers in the paramedian lobule of the cerebellum in children. Material and methods. The work was performed on postmortem material (62 cerebellums) obtained from children aged from birth to 12 years who died from injuries but without brain damage. The thickness of the cortex, as well as the thickness of its molecular and granular layers, were measured using computer morphometry on the Nissl-stained frontal histological sections of the cortex taken bilaterally in the region of the paramedian (gracile) lobule (HVIIB) at the top of the folia of cerebellum. Analysis of quantitative data was performed at annual intervals. Results. In the paramedian lobule of the cerebellum, the increase in the thickness of the cortex occured in four stages: in the right hemisphere - from birth to 1, 3, 5 and 9 years, in the left hemisphere - to 1, 5, 7 and 9 years. Left-sided asymmetry of the cortical thickness of the cerebellum was observed in 1 and 2-year old children, the thickness of the molecular layer - in 3-year old children. Right-sided asymmetry was characteristic for the thickness of the granular layer in 3-year old children and a cross-section of the cortex in 6-year old children. The thickness of the cortex and layers in the area of the paramedian lobule of the cerebellum on the average group indicators reached the level of adults by 9 years. Conclusions. The thickness of the cerebellar cortex and its layers in the area of the lobule HVIIB increased heterochronically and heterogeneously in the right and left hemispheres of the cerebellum in children of the first year of life, and in the periods of early, first and second childhood. No reduction in the diameter of the cortex and layers in the paramedian lobule of the cerebellum of children from birth to 12 years was found.

Immunohistochemical profile of mononuclear infiltrate in the myocardium of transplanted heart. Computer morphometry data

Aim. To carry out a quantitative immunophenotypic characterisation of cellular corporations in a mononuclear inflammatory myocardial infiltrate in the cell and humoral forms of heart transplant rejection using the computer morphometry of endomyocardial biopsy samples.Materials and methods. Endomyocardial biopsy samples (n = 226) were obtained from 56 heart recipients who underwent transplantation in 2018–2019. Sections with a thickness of 5 μm after the paraffin infiltration were stained with hematoxylin and eosin according to the standard procedure. The expression of CD3 T-lymphocyte, CD20 B-lymphocyte and CD68 macrophage markers was determined by the immunohistochemical streptavidin-biotin method. Using computer morphometry, the staining area coefficient (SAC) was calculated as the percentage of the total area of the stained objects to the area of the biopsy. The statistical processing of the results included verification of the distribution nature by the Kolmogorov-Smirnov method and the calculation of the Cramer — Welch criterion.Results. Lymphocytes and macrophages were found in the inflammatory infiltrate of all heart transplants. The expression of CD3 T-lymphocyte marker in the absence of rejection (0R) was at SAC = 0.99 ± 0.02%. In comparison with 0R cases, this coefficient increased 2.1 times (p <0.05), 3.4 times (p <0.05), 5.5 times (p <0.05) and 4.8 times (p <0.05) in 1R, 2R, 3R and humoral rejection, respectively. The expression of the CD20 B-lymphocyte marker in 0R cases was characterised by SAC = 0.19 ± 0.01%. In comparison with 0R cases, this coefficient increased 2.7 times (p <0.05), 3.4 times (p < 0.05), 4.4 times (p <0.05) and 9.5 times (p <0.05) in 1R, 2R, 3R and humoral rejection, respectively. The value of the CD68-positive macrophage region for 0R was only 0.34 ± 0.01%. This parameter increased 2.7 times (p <0.05), 4.0 times (p <0.05), 9.6 times (p <0.05) and 4.1 times (p <0.05) in 1R, 2R, 3R and humoral rejection, respectively.Conclusion. Cellular corporations in the mononuclear inflammatory infiltrate of transplanted heart are characterised by the predominance of T-lymphocytes in the cases of both cellular and humoral rejection. The expression of the B-lymphocyte marker is most pronounced in an antibody-mediated form. The maximum presence of macrophages in the infiltrate characterises severe cell rejection. An increase in the severity of cell rejection leads to an increase in the relative content of B-lymphocytes and macrophages in the infiltrate.

The key role of pancreatic fibrosis severity in the surgical treatment algorithm of patients with chronic pancreatitis

The aim of the work was to improve the results of surgical treatment of complicated forms of chronic pancreatitis. Materials and methods: The results of surgical treatment of 181 patients with complicated forms of chronic pancreatitis have been analyzed. All these patients were treated in surgical clinic of 2 nd medical faculty of National Pyrogov Memorial Medical University in Vinnytsya. Results: It is possible preoperative indirectly assess the severity of fibro- degenerative changes in pancreas, that includes 1 – the definition of pain type; 2 – CT or MRI (Marseilles- Rome classification); 3 – assessment of the stage of chronic pancreatitis (Büchler classification); 4 – patient&#39;s nutrition status and preoperative differential diagnosis with pancreatic cancer; 5 – assessment of the fibrosis severity (elastomers). The key point in treatment depended on intraoperative examination: detection of strictures of the main pancreatic duct (pacemaker of chronic pancreatitis); the tissue pressure resistance to the liquid, which is the maximum value&gt; 200 mmHg, in the region of stricture and falls in other parts of the pancreas; pressure in the main pancreas duct, which rises only in 59.5% of patients. The head of the pancreas was involved in the pathological process in 83.8%, in 16.2% it was isolated in the isthmus or the body and tail of the pancreas. In general, the distal pancreas was involved in 37.8%. At computer morphometry of histological samples, the area of connective tissue fields reached 81.4 ± 6.62%, preserved exocrine part in 4.87 ± 1.62%, endocrine – 1.92 ± 0.12%, total area of ducts – 6 , 47 ± 1.12%. Conclusions: The combined Frey-Izbickiy local resection provides a wide excision of the pacemaker (stricture) of chronic pancreatitis. In case of extrapancreatic complications or repeated surgical interventions on the pancreas due to chronic pancreatitis, this effect can be achieved by pancreatoduodenal resection.

Cytoprotective effect of non-opioid leu-enkephalin analogue in primary culture of pulmonary fibroblasts in oxidative stress

Aim. Analysis of the cytoprotective effect of non-opioid leu-enkephalin analogue (Phe-D-Ala-Gly-Phe-Leu-Arg) in the primary culture of pulmonary fibroblasts in conditions of oxidative stress. Methods. Pulmonary fibroblasts were incubated with the peptide of non-opioid leu-enkephalin analogue (Phe-D-Ala-Gly-Phe-Leu-Arg) in the concentration 0.1 μM for 6 hours. To simulate oxidative stress, 60 μM H2O2 was added to the culture medium for 2 hours. Experimental series included (1) «control»; (2) «non-opiate leu-enkephalin analogue» (the peptide was added to the culture medium 44 hours after the final passage); (3) «oxidative stress» (H2O2 was added to the culture medium 48 hours after the final passage); (4) «non-opiate leu-enkephalin analogue + oxidative stress» (the peptide and H2O2 were added to the culture medium 44 and 48 hours respectively after the final passage). In order to evaluate the generation of superoxide anion by pulmonary fibroblasts, the method of lucigenin-dependent chemiluminescence was used. Computer morphometry of the nucleo-nucleolar apparatus of fibroblasts stained with silver nitrate by the AgNOR method was used to assess the cell state: the area of fibroblast nuclei, the total nucleoli area in the nucleus, and the number of nucleoli in the nucleus were measured. These parameters correlate with the activity of anabolic processes in the cells. Results. The effect of H2O2 on the primary culture of pulmonary fibroblasts caused an increase of superoxide anion generation by the fibroblasts, reduction of fibroblast nuclei size, decrease of nucleoli amount and size. Pre-incubation of pulmonary fibroblasts with a non-opioid leu-enkephalin analogue reduced the H2O2-induced generation of superoxide anion, corrected changes in the nucleo-nucleolar apparatus of fibroblasts caused by oxidative stress. In our previous studies, similar effect in the same model was shown for non-selective μ/δ-opioid receptor agonist peptide sedatin (dermorphin analogue). The mechanism of cytoprotective effect of non-opioid leu-enkephalin analogue may include the affinity of this peptide to nociceptin receptors (NOR receptors) that requires further studies. Conclusion. The results of the study indicate a direct cytoprotective effect of the peptide Phe-D-Ala-Gly-Phe-Leu-Arg (non-opioid leu-enkephalin analogue) in oxidative stress.

Ligature induced periodontitis causes atherosclerosis in rat descending aorta: an experimental study

Introduction. In recent years a link between periodontitis and cardiovascular diseases has been proposed. Oxidative stress has been found to be involved in their pathophysiology. Objectives. The aim of this study was to demonstrate that a wire ligature placed around the first mandibular molars of the rat causes inflammation in the periodontium and lipid deposits in the aorta, via oxidative stress. Methods. Twenty male Wistar rats were divided randomly into two groups of 10: Control group and Periodontitis group. An orthodontic wire ligature was placed around the cervix of the first mandibular molar and was kept in place for 4 weeks. Clinical attachment loss was determined to assess the gingival recession. Aorta was harvested after 4 weeks and the malondialdehyde (MDA) levels in the aorta tissue homogenate were determined. Lipid deposits were evaluated using Oil Red staining and immunofluorescence under confocal microscopy. Total lipid deposits in the aorta were assessed using a computer morphometry system. Results. Significant gingival recession was observed in the Periodontitis group in comparison with the Control group. Increased MDA levels were present in the aorta in the ligatured rats in comparison with the Control group. Significant lipid deposits were observed in the aorta of the Periodontitis group as opposed to the Control group. Conclusions. The affixing of an orthodontic wire ligature produces gingival recession, increased oxidative stress and lipid deposits in the rat descending aorta.

The Non-Invasive System to Control Morphogenesis of Planaria as an Alternative Approach to Bioassessment

The published study tests our own chemical luminescence Planaria accompanying regeneration processes in combination with methods of in vivo fixation of their structure using non-invasive computer morphometry. As a result, we propose a new system of online remote control of morphogenesis in Planaria regeneration. This system is promising for primary screening of drugs – proliferation regulators.