Mallory's Method Research Articles

Morphological characteristics of the stromal-parenchymatous component of the liver of fetal and newborn developed under the conditions of maternal hypertension disease

Background. Extragenital pathology of the mother can cause various changes in the body of the newborn, which in the future are the cause of violations in physical and mental development, and also causes an increase in somatic and infectious diseases in such children. Therefore, timely diagnosis of liver diseases in newborns and young children remains an urgent problem today. Purpose – the article is devoted to the study of the morphological features of the stromal-parenchymal component of the liver of fetuses and newborns that developed under conditions of maternal hypertension. Materials and Methods. Research was conducted on liver preparations of fetuses and newborns who died at the age of 37 weeks of gestation – the 1st day of postnatal life. All the studied material was divided into two groups. The control group (group K) included fetuses and newborns from healthy mothers (18 cases) who died due to acute disruption of umbilical-placental circulation and birth trauma. The comparison group (group C) consisted of 36 fetuses and newborns from mothers with stage II hypertension. The obtained micropreparations were stained with hematoxylin and eosin, picrofuchsin according to the van Gizon method, and according to the Mallory method. The parenchymal, stromal, and vascular components were calculated as a percentage, and then the stromal-parenchymal ratio was calculated. Results. The results we obtained during the conducted research indicated that the influence of chronic intrauterine hypoxia, which developed against the background of stage II hypertensive disease in the mother, leads to the development of destructive-dystrophic changes in the liver tissue of fetuses and newborns. Chronic intrauterine hypoxia with maternal arterial hypertension leads to parenchymal losses and has a pronounced stimulating effect on the stromal component of the liver, and this can lead to a significant decrease in the functional activity of the liver under the influence of certain triggers and increases the risk of developing liver failure and liver cirrhosis in the future. Conclusions. During our research, it was established that chronic intrauterine hypoxia, which develops against the background of stage II hypertensive disease in the mother, leads to the development of significant destructive-dystrophic changes in the liver tissue of fetuses and newborns. There is a decrease in the volume of the parenchyma due to the volume of hepatocytes and an increase in the volume of the stroma with vessels due to the volume of the portal tracts (11.2 ± 2.2% compared to the control – 3.4 ± 0.3%), the volume of the central veins (12.9 ± 1.3% compared to the control – 9.4 ± 0.4%), the volume of sinusoids (13.3 ± 1.1% compared to the control – 11.3 ± 0.8%). It was established that the morphological changes described by us in the liver of fetuses and newborns in the subsequent ontogenesis may become a prerequisite for the development of hepatobiliary system pathology in such children.

Role of CD68+ and CD206+ cells in the progression of toxic liver fibrosis in rats

The aim of the work was to evaluate the role of stellate macrophages in a large number of points of toxic liver fibrosis in rats. Liver fibrosis and cirrhosis in male Wistar rats were induced with thioacetamide at a dose of 200 mg/kg animal weight for 17 weeks. Histological preparations of the liver were stained with hematoxylin and eosin according to the Mallory method. Immunohistochemical examination was performed on paraffin sections using monoclonal mouse antibodies CD68 and polyclonal rabbit antibodies CD206. The fibrosis degree was determined according to the Ishak semi-quantitative scale. Toxic liver fibrosis before the start of its transformation into cirrhosis (9 weeks) was accompanied by an increase in the number of CD68+ cells compared with the control. At all subsequent experiment stages, no differences were found in comparison to the control. In the liver of control rats, CD206+ cells were practically absent. Throughout the experiment, their number remained above the control point – 3 weeks. With the progression of liver cirrhosis, a decrease in the number of CD206+ cells was noted, but it did not reach a level of 3 weeks. Morphologically, two different groups of CD68+ cells were identified. One group of cells had a pterygoid shape and they were located mainly in the liver sinusoids. The second group of CD68+ cells had a round shape and different localization. They were detected around the vessels of portal zones, surrounded brown pigment accumulations in connective tissue septa, were observed near single lying groups or groups of giant hepatocytes and liver cells containing brown pigment in the cytoplasm, and were also noted in the foci of necrosis of hepatocytes. Cells, expressing the CD206 marker, are round in shape and are elongated and located in the liver sinusoids. Presumably, round-shaped CD68+ cells perform a phagocytic function, and pterygoid-shaped CD68+ cells transdifferentiate into CD206+ cells that have anti-inflammatory properties.

THE MORPHOLOGICAL AND FUNCTIONAL ASSESSMENT OF FAP+ AND α-SMA+-CELLS IN DIFFERENT TIMES OF RAT TOXIC LIVER FIBROSIS

Qualitative study of the source of the fibro-genic cell population in relation to the etiology and stage of fibrosis, as well as an understanding of the molecular mechanisms that regulate changes in the phenotype of hepatic fibroblasts, are of paramount importance in the development of pharmacological drugs. The purpose of the study was a morphological and functional assessment of activated portal fibroblasts (FAP+) and fat-accumulating cells (α-SMA+) of the liver at various stages of toxic liver fibrosis in rats. Liver fibrosis and cirrhosis in male Wistar rats were induced with thioacetamide solution for 17 weeks. Morphological examination of the liver was carried out on paraffin sections stained with hematoxylin and eosin using the Mallory method; immunohistochemical examination was carried out using polyclonal rabbit antibodies to the portal fibroblast antigen FAP and using monoclonal mouse antibodies to the α-SMA+ cell antigen. Before the onset of liver fibrosis stage F3/F4, from weeks 3 to 7, the number of FAP+ and α-SMA+ cells increased alternately. During the stages of transformation of fibrosis into cirrhosis from 7 to 11 weeks, their number increased slightly. At the stage of incomplete (F5) and before the onset of significant cirrhosis (F6) from weeks 11 to 15, the number of FAP+ and α-SMA+ cells were inconsistent and there was an alternating increase and decrease in their number. α-SMA+ cells before the start of the process of transformation of fibrosis into cirrhosis (F4/F5) were observed in sinusoids and foci of necrosis. Then they were detected both in sinusoids and in connective tissue trabeculae. FAP+ cells at the stage of portal fibrosis (F1) were localized near the interlobular vessels and interlobular bile ducts of the portal zones, and from the F2/F3 period they were detected in connective tissue trabeculae and sinusoids. In quantitative terms, α-SMA+ cells predominated at all stages of fibrosis. Based on the results obtained, it can be assumed that FAP+ cells make a major contribution to the development of the portal and initial stages of bridging fibrosis. They should be considered as one of the myofibroblast populations in thioacetamide-induced liver fibrogenesis.

Уровень мРНК гена Nos2 как маркер фиброгенеза печени крыс, индуцированного тиоацетамидом

The aim of this paper was to evaluate the potential of the mRNA level of the Nos2 gene as a marker of fibrogenesis in rats at different stages of thioacetamide-induced liver fibrosis and cirrhosis. Materials and methods. The experiment involved 117 mature male Wistar rats weighing 190–210 g. Liver fibrosis and cirrhosis were induced by a thioacetamide solution administered through a gastric catheter at a dose of 200 mg per 1 kg of body weight 2 times a week. The dynamics of the process was studied at 9 timepoints over the course of 17 weeks. Nos2 mRNA level in the liver was detected by means of real-time polymerase chain reaction. The stage of fibrosis was determined in histological sections stained using the Mallory method according to the Ishak semi-quantitative scale. Results. Throughout the experiment, Nos2 mRNA had practically no reaction to the thioacetamide-induced liver fibrogenesis. At the mild fibrosis stage (F1), an insignificant rise in the level of Nos2 mRNA was noted (within 5 %). Intensive development of fibrosis (F2–F4/F5) and an increase in the production of extracellular matrix components were accompanied by an increase in the level of Nos2 mRNA with a peak value exceeding the initial level (control group value) by the factor of 1.69 (р < 0.05). At the point of transition of fibrosis to cirrhosis (F5), a decrease in the level of mRNA of the target gene was observed, and at the stage of definite cirrhosis (F6), a subsequent drop below the initial level was detected. Thus, according to the data obtained, Nos2 mRNA has the greatest potential as a marker of liver fibrogenesis at the stage of advanced fibrosis, but cannot act as a marker at the early stages. Moreover, Nos2 mRNA level cannot be used as a marker when assessing the degree of cirrhosis and its development dynamics.

EVALUATION OF DIAGNOSTIC ALGORITHM BASED ON COLLAGEN ORGANIZATION PARAMETERS FOR BREAST TUMORS.

Tumor tissue samples of 5patients with breast fibroadenomas and 20patients with stage I-II BCa were studied. Collagen was identified histochemically by Mallory method. Photomicrographs of thestudied preparations were obtained using a digital microscopy complex AxioScope A1. Morphometric studies were performed using thesoftware CurveAlign v. 4.0. beta and ImageJ. Thealgorithm for determining thequantitative characteristics and spatial organization of thecollagen matrix in tumor tissue samples has been developed and tested. We showed that collagen fibers in theBCa tissue are characterized by significantly lower values of length (p < 0.001) and width (p < 0.001) as well as higher values of straightness (p < 0.001) and angle (p < 0.05) compared to these in thefibroadenoma tissue. No significant difference was found in thedensity of collagen fibers in thetissue of benign and malignant neoplasms of themammary gland. Thealgorithm allows assessing a wide range of parameters of collagen fibers in tumor tissue, including their spatial orientation and mutual arrangement, parametric characteristics and density of thethree-dimensional fibrillar network.

Stereological analysis of vascular network of subcortical auditory centers

&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Introduction. &amp;lt;/strong&amp;gt;Subcortical auditory centers have several sources of blood supply. Cochlear nuclei are vascularized by the anterior inferior cerebellar artery. Superior cerebellar and posterior cerebral arteries supply the inferior colliculus nucleus, while the medial geniculate body nucleus is supplied by several posterior cerebral artery branches. The aim of the research was to quantify the vascular network of subcortical auditory centers.&amp;amp;nbsp;&amp;lt;/p&amp;gt;&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Methods.&amp;lt;/strong&amp;gt; Twelve adult brains, both sexes, aged 21 to 78 years, without signs of neurological diseases were analyzed in this study. Samples for histological sections, stained by the Mallory method, were obtained from strata cut at levels: the junction of the middle and rostral thirds of the oliva, the inferior colliculi, and the thalamic pulvinar. Volume, surface, and length density of the vascular network of subcortical auditory centers were analyzed stereological parameters.&amp;amp;nbsp;&amp;lt;/p&amp;gt;&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Results.&amp;lt;/strong&amp;gt; All parameters had the highest values in the medial geniculate body nucleus. Statistically significant difference was found in the volume, surface, and length density between vascular networks of the cochlear nuclei and medial geniculate body nucleus, and between inferior colliculus nuclei and medial geniculate body. Vessels in cochlear nuclei and inferior colliculus nuclei differed only in length density.&amp;amp;nbsp;&amp;lt;/p&amp;gt;&amp;lt;p&amp;gt;&amp;lt;strong&amp;gt;Conclusion.&amp;lt;/strong&amp;gt; Cochlear nuclei and inferior colliculi nuclei blood vessels have a similar density and diameter, but vessels in inferior colliculi nuclei are more tortuous. In the medial geniculate body nucleus vessels are larger, denser and more tortuous compared to the other two subcortical auditory centers&amp;lt;/p&amp;gt;

Morphometric parameters of sinusoidal capillaries, interlobular veins and interlobular arteries at different stages of experimental liver fibrosis

The aim of the study was to conduct a morphometric analysis of sinusoidal capillaries, interlobular veins and interlobular arteries of the liver at different stages of experimental fibrosis in order to investigate relationship between fibrogenesis and angiogenesis and identify new therapeutic targets. Material and methods. The study included male Wistar rats. Liver fibrosis with transformation into cirrhosis was induced with a solution of thioacetamide, which was injected into the stomach, dosage 200 mg/kg of animal body weight, using a probe 2 times a week. The rats of the experimental groups were withdrawn from the experiment in 3, 5, 7, 9, 11, 13, 15, and 17 weeks, and the intact ones - at the end of the experiment. To identify connective tissue, liver sections were stained according to the Mallory method. Immunohistochemical examination was performed on paraffin sections using a monoclonal mouse antibody to CD31. The number of sinusoidal capillaries, interlobular arteries, and interlobular veins was counted in three non-overlapping fields of view of each histological section using an OLYMPUS BX51 microscope at a 40× objective magnification. Results. The results obtained supported a statistically significant decreased number of sinusoidal capillaries (p=0,0000) and an increased number of interlobular veins (p=0,0000) in animals of the experimental group compared with the control group. There were no differences in the number of interlobular arteries (p=0,9999). Pronounced angiogenesis in the portal zones and connective tissue septa was associated with an increased amount of the connective tissue during liver fibrogenesis. Conclusion. In the future, angiogenesis modulation should be differentiated and directed towards antifibrotic, anti-inflammatory or autoimmune therapy in chronic liver diseases.

Digestive system of the hepatotrematode &lt;i&gt;Parafasciolopsis fasciolaemorpha&lt;/i&gt; Ejsmont, 1932 (Plathelminthes, Trematoda): morphofunctional organization, histological and histochemical features to adaptive specialization

The purpose of the research is to study the morphofunctional organization, histological and histochemical features of the digestive system of the marita Parafasciolopsis fasciolaemorpha, which provide adaptation to parasitism in the endostasis – the liver Alces alces.Materials and methods. Mature specimens of the trematode P. fasciolaemorpha (Ejsmont, 1932) collected from the bile ducts of the Alces alces liver served as the material. Maritas were fixed in 70 and 80% alcohols, Schaffer alcohol-formalin 1 : 9, and 10% neutral formalin. Histological stains: with hematoxylin-eosin and by the Mallory method, followed by additional staining of the nuclei with Orta lithium carmine. Histochemical stains: sublimate-bromophenol blue according to Bonheg, Schick reaction according to McManus with additional staining of nuclei with Mayer's hemalaune, alcian blue according to Steedman and Mowry (pH 3.0 and 2.2) and toluidine blue (pH 2.0-5.0).Results and discussion. Parafasciolopsis fasciolaemorpha (Ejsmont, 1932) is the most pathogenic hepatoparasite of Alces alces L. The study of the features of the morphological organization of the digestive system as one of the border systems of the organism of the parasite revealed a number of features: the muscles of the large oral sucker contain many total proteins, which is confirmed by intense bromophenolophilia; in the thickness of the sucker wall there are small secretory cells and neurosecretory cells with abundant alcian- and toluidinophilic vacuolated cytoplasm; the presence of single and grouped digestive glands located in the parenchyma at the junction of the prepharynx to the pharynx, and on the border between the pharynx and the esophagus; in the apical part of the intestinal epithelium, a “brush border” is formed, the thin microvilli of which contribute to an increase in the working surface of parietal digestion in P. fasciolaemorpha, enhancing the trophic processes of the helminth to survive in the body owner. The revealed histological and histochemical features of the parafasciolopsis digestive system can be considered as examples of adaptive specialization at the site of endostasis.

Pathomorphological peculiarities of liver of fetuses and newborns born in conditions of hypertensive disease in the mother

Aim. The article is devoted to the study of pathomorphological features of fetal liver and newborns affected by chronic intrauterine hypoxia, which developed against maternal hypertension. Materials and methods. The studies were performed on liver preparations of 18 fetuses and newborns from mothers with physiological pregnancy (group I) and 36 fetuses and newborns from mothers with stage II hypertension (group II) at the age of 37 weeks of gestation - the 1st day of postnatal life. In each case, three fragments were excised from the liver, and then the material was fixed in 10 % formalin solution. Formalin-fixed tissue compaction was achieved by conducting, through alcohols, increasing concentrations, celloidin, chloroform, and paraffin embedding. From the prepared blocks for serial color serial sections with a thickness of 4–5 microns were prepared. Micropreparations were stained with hematoxylin and eosin, picrofuxin by the van Gizon method, and by the Mallory method. The diameter of the hepatocytes and their nuclei, NPP, were calculated, and the number of single and double-nucleated hepatocytes was calculated. The material was studied on an «Olympus BX-41» microscope with a digital camera. Results. The results of our study showed that the influence of chronic intrauterine hypoxia on the background of stage II hypertension led to the development of destructive-dystrophic changes in the liver tissue of the fetus and newborn. Structural and functional disorders, in turn, led to the activation of cellular regeneration mechanisms. In group II, the recovery of structural and functional integrity of the liver occurred due to an increase in the number of double-nucleated hepatocytes and manifested by an increase in their number almost 2-fold. Conclusions. During the study of pathomorphological features of fetal liver and newborns it was found that stage II hypertension in the mother leads to the development of significant destructive-dystrophic changes in the tissue of fetal liver and newborns. The diameter of hepatocytes of fetuses and newborns of group II was significantly increased compared with group I (24.3 ± 2.4 μm and 18.71 ± 2.8 μm, respectively). The number of hepatocytes in the field of view in the fetuses and newborns of group II significantly decreased compared with group I (214.8 ± 22.80 and 268.1 ± 24.11 cells, respectively). The ratio of the number of binucleate hepatocytes to the number of binucleate hepatocytes in fetuses and newborns of group II compared to group I increased 2.5 times (0.08 ± 0.02 and 0.03 ± 0.02, respectively).

Stereological analysis of the vascular network in superior colliculi

Background/Aim: Superior colliculi have an important role in the visual system, some motor functions and behaviour. Branches of the posterior cerebral artery and surrounding vessels form a collicular arterial network, which gives rise to several penetrating vessels for superior colliculi. The aim of the study was to determine relative stereological parameters (volume, surface and length density) of the vascular network in superior colliculi and to correlate those parameters with age. Methods: Ten adult brainstems, both male and female, were cut transversally at the middle of superior colliculi in three-millimetre-thick strata. Every stratum was cut in 4 µm thick semiserial sections stained with Mallory method. Images of every fifth section (5, 10, 15, 20) of superior colliculus were taken with "Leica" DM 1000 microscope and "Leica" MC 170HD camera under x 400 magnification and analysed by ImageJ software with A 100 grid. Statistical analysis was performed by the SPSS software with 5 % level of significance using methods of descriptive statistics, Spearman's and Pearson's correlation coefficient. Results: The average volume density of vascular network in superior colliculi was 0.0203 mm 0 , surface density 6.133 mm-1 , and length density 205.357 mm-2. All those parameters decreased with ageing, but statistically significant change was seen only in length density (p = 0.006). Conclusion: Compared to the structures at the same transversal level, the vascular network of superior colliculi has an average density. With ageing, blood vessels become less tortuous.

Sensitivity of life-long diagnostic methods for geese nematodes

The basis of successful struggle and specific prevention of poultry worms is timely diagnosis, the final stage of which is the detection of the worms themselves, their eggs or larvae at various stages of development. Priority is given to the methods of lifelong laboratory diagnosis of helminthiasis, which are preferably recommended for use in all animal species, including poultry. The aim of this work was to determine the sensitivity of flotation methods of coprovoscopy for geese nematodes. Experimental determination of the efficiency of the well-known methods of flotation and their comparative evaluation of coproovoscopic diagnosis of heterocosis, capillary disease and trichostrongilosis of geese. The main indicator of the diagnostic effectiveness of the methods was the intensity of the invasion and the time spent on the flotation of the samples. The most effective methods for diagnosing geese geoccosis are Kotelnikov-Hrenov (with ammonium nitrate) – at exposures of 20 min and Mallory (with saturated sugar solution) – at exposures of 10–15 min. The rates of invasion intensity were respectively 62.0 ± 4.39 and 59.0 ± 3.47 eggs/g. In the laboratory diagnosis of goose capillary disease, the most sensitive methods were Kotelnikov-Hrenov and Mallory at exposures of 15–20 min, where infestation rates reached 34.0 ± 2.22 and 33.5 ± 2.64 eggs/g, respectively. For trichostrongilosis, the Kotelnikov-Hrenov method showed the highest sensitivity at the exposure of 20 min, the intensity of the invasion was 32.5 ± 3.23 eggs/g. The Mallory method proved to be less effective – at an exposure of 15 min poultry invasiveness was 23.5 ± 1.81 eggs/g. The least sensitive of this invasion was the Fulleborn method (with NaCl), where the intensity of the invasion gradually increased with prolonged exposure and ranged from 10.5 ± 0.5 to 19.5 ± 2.45 eggs/g. Based on the data obtained, it is recommended to use the most sensitive methods and to take into account the exposure, which ensures the concentration of the largest number of nematode eggs on the surface of the flotant when conducting life-long coproovoscopic diagnostics of heterosis, capillary disease and trichostrongilosis of geese.

Experimental and morphological substantitation of application of PRP-technology for endoprosthesis of anterior abdominal wall

In modern practical health care a large aspect is occupied by PRP-technology, the which effect is provided by growth factors located in alpha granules on the platelet membrane and responsible for the regeneration of connective tissue. The purpose of this study was a morphological assessment of reactive changes in periprosthetic tissue in PRP-technology application. Material and methods . The study was carried out on laboratory rabbits. Experimental animals were divided into two groups of observations - control and experimental. The endoprosthesis was implanted into the anterior region of the abdominal wall. Autoplasma enriched with platelets was injected after implantation under the endoprosthesis twice into experimental animals. A site with an endoprosthesis material was excised for a morphological study. Histological sections were stained with hematoxylin and eosin, according to Van Gieson, and Mallory methods, an immunohistochemical study was performed. The results of the morphometric study were processed using the Statistica v. 8.0. Results. In the experimental group the onset of the proliferative phase of inflammation occurred at an earlier time and provided the formation of a mature connective tissue periprosthetic capsule. By the 10th day of the experiment, a double dominance of the thickness of the capsule was observed in the experimental group. The proportion of cells of fibroblastic differon accounted for 60% of all cells, which resulted in an an increase in the cellular index by 1.6 times. The proliferation index was 2 times greater at all experimental days. Conclusion. The use of PRP-technology in endoprosthetics leads to an earlier formation of a mature periprosthetic capsule, accelerates the survival of the prosthesis and reduces the risk of postoperative complications.

Age- Related Changes of Quantitative Parameters of Neurons in Extraocular Motor Nuclei

Introduction: Extraocular motor nuclei are located in the midbrain (principal oculomotor and trochlear nucleus) and in the pons (abducens nucleus). With aging, there are significant changes in eyeball mobility. Aim of the Study: The aim was to determine whether the quantitative parameters of neurons (volume and surface density, and the absolute number per mm² of the surface) in these nuclei significantly change with aging. Patients and Methods: The study was done on 30 adult brainstems, both male and female, without diagnosed neurological disturbances. Three-millimeter-thick strata were taken in transversal plane and cut in 0.3 micrometer semi-serial sections stained with Mallory method. The images of studied nuclei were taken by 'Leica' DM 1000 microscope and 'Leica' EC3 digital camera under 400x magnification and analyzed by ImageJ software with A100 grid. The statistical analysis was performed by Statistical Package for the Social Sciences software using Pearson's correlation coefficient with 5% level of significance. Results: The volume density of neurons had highly statistically significantly increased with age in principal oculomotor nucleus (r = 0.571, p = 0.001) and trochlear nucleus (r = 0.581, p = 0.001), while abducens nucleus showed no change in neuron volume. Changes of values of surface density and absolute number of neurons per mm2 with age did not reach statistical significance. Conclusion: Volume of neurons of extraocular motor nuclei located in the midbrain increase with age, while their surfaces and absolute number do not change significantly. These changes are not observed in the nucleus located in the pons.

Changes of Neurons and Blood Vessels of Human Substantia Nigra in Aging- Morphometric Study

Introduction: With aging, populations of dopaminergic neurons in the central nervous system show prominent pathological changes compared to other brain regions. Previous studies on substantia nigra were performed in cases of Parkinson's disease and in old age. Aim of the Study: Since Parkinson's disease is a disorder associated with age, it is important to examine how the relationship between neurons and blood vessels is associated with normal aging. Patients and Methods: Ten brainstems were sliced into three strata. Each stratum was sliced in semiserial sections and stained by Mallory method. Studied phases were neurons and blood vessels of substantia nigra. The analysis was conducted by camera 'Leica EC3' under the 40x magnification of light microscope 'Leica' DM 1000, using ImageJ software (version 1.42 e). Determined morphometric parameters of neurons and blood vessels were: volume and surface density, and absolute numbers per visual field. Statistical analysis was performed using SPSS software, version 16.0, using Student's t-test and Pearson correlation coefficient. Results: Volume and surface density, and total number of neurons per visual field of substantia nigra significantly decreased with age, while the volume and surface density and absolute number of blood vessels per visual field significantly increased (p < 0.05 ). Conclusion: Decrease in size and number of neurons occurs with aging, which is compensated by the increase of vascular network. This affects the supply of nutrients from the blood to neurons, as well as the availability of blood cells or toxic substances, but also the susceptibility to neuronal diseases.

Endometrial dysfunction caused by chronic endometritis: сlinical and morphological aspects

to determine the clinical and morphological aspects of endometrial dysfunction caused by chronic endometritis (CE). The study included 239 reproductive-aged patients: 93 were examined for abnormal uterine bleeding; 37 patients of them had a history of miscarriage and secondary infertility, 17 patients had primary infertility (Group 1). The remaining 105 patients with infertility were examined for future in vitro fertilization (Group 2). A comparison group consisted of 41 patients with normal menstrual cycles and reproductive function across the cycle, who had separate diagnostic curettage before forthcoming surgery for uterine myoma. All the women in Groups 1 and 2 underwent standard clinical and morphological investigations, histopathological examination of the material obtained during hysteroscopy with separate diagnostic curettage (Group 1), and aspiration biopsy (Group 2) with the sections being stained with hematoxylin and eosin and by Mallory's method. Immunohistochemical diagnosis was used to assess endometrial receptivity to steroid hormones and glycodelin. Antibodies against CD4, СD8, and CD20 were employed to study local immunity. In Group 1, 52 (55.9%) of the 93 patients admitted to hospital for abnormal uterine bleeding were found to have CE on the basis of a morphopathological examination of the material after separate diagnostic study. In Group 2, CE was established in 59 (56.9%) of the 105 patients after postmortem examination. The patients with CE had a history of reproductive significant infections, such as Chlamydia, Trichomonas, Ureaplasma Mycoplasma, cytomegalovirus, papillomavirus infections, genital herpes, genital candidiasis, and bacterial vaginosis. The CE groups showed functional disorders of the endometrial glandular and surface epithelium. In the middle stage of proliferation, the expression of glycodelin in the glandular epithelial cells was detected to be moderate to strong in 83.3%. During the periovulatory period, no secretion of glycodelin is of fundamental importance for the regulation of reproductive function because this protein has contraceptive activity, by blocking the binding of sperm to the zona pellucida. Accordingly, endometrial glycodelin production during the proliferative phase, which has been identified by the authors in patients with CE, may be one of the pathogenic mechanisms for the development of infertility. In addition, the patients in Groups 1 and 2 compared with those in the comparison group were noted to have a decrease in CD4 cell counts with a simultaneous increase in CD8 expression and a reduction in CD20 levels, especially in Group 2. The patients with SE were identified to have endometrial dysfunction characterized by lower reception to steroid hormones, impairment in glycodelin secretion, retardation in the development of pinopods and in the phase of the menstrual cycle, and local immunity disorders. The above endometrial changes should be taken into account in the pregravid preparation of patients with CE.

Features of cytotrophoblast invasion in complete placenta previa and increta

to investigate the characteristics of cytotrophoblast invasion in complete placenta previa and increta. Three groups of placentas and amputated uteri were examined. These were: 1) 10 placentas at 20-22 weeks' gestation after drug-induced abortion; 2) 4 uteri with typical placentation at 34-36 weeks and wall ruptures; 3) 12 uteri with ultrasound-confirmed complete placenta previa and subsequent hysterectomy (at 34-36 weeks.) due to massive bleeding. In all cases, the sections were stained with hematoxylin and eosin, azan by the Mallory's method; immunovisualization of invasive cells with the marker cytokeratin 8 was also used. In Groups 2 and 3, the uterine distribution density of invasive cells was compared in a standard slice area (×200) separately, within the endometrium and myometrium. Complete placenta previa was found to have the following characteristics: 1) all the uteri exhibited focal or diffuse friable, or thick scars after cesarean section; 2) multiple active anchor villi with villous cytotrophoblast layers, which were characteristic of Group 1 placentas and absent in the uteri women of Group 2; 3) bays diagnosed in the basal endometrium with ingrown villi (placenta increta); 4) a morphometrically significant increase in the distribution density of interstitial cytotrophoblast in the endometrium and only a similar trend in the myometrium. Invasive cells did not penetrate into the area of scars. Failure of the second wave of cytotrophoblast invasion was confirmed by incomplete gestational restructuring and partial obliteration of the myometrial radial arteries. Real risks for severe clinical forms of abnormal placentation declare more stringent indications for surgical delivery.

Influence of mother HIV infection on pancreas development of the fetus.

Background. Current stage of HIV infection is characterized by a significant increase in the incidence of infection among women of reproductive age and pregnant women. Consequently it will lead to the complex of problems related to the health of children born to them. Objective. To identify the impact of HIV infection of the mother on the formation of the pancreas of the fetus. Methods. Pancreases were measured and weighed. Histological sections were stained with hematoxylin and eosin, Van Gison and Mallory methods. Immunohistochemical examination was performed using the indirect method of Coons in modifications of M.Brosman. α- and β-cells were detected using monoclonal antibodies to glucagon and insulin. Immunohistochemical study was carried out in the luminescent microscope «Axioskor 40" using the software Biostat.exe. All digital data is processed by methods of mathematical statistics using variations, alternative and correlation analysis. Results. Analyzing the morphometric parameters of the pancreas of stillbirths from HIV-infected mothers, comparing them with the control group we found a massive expansion of the stroma on the background of decreasing both quantitative and qualitative composition of the islets of Langerhans, as well as a violation of the ratio of populations α- and β-cells in the direction of decreasing β-endocrine cells (1: 1.47). The latter ratio is known to play a leading role in the development of the endocrine pancreas dysfunction. Decrease in the density of Langerhans islets due to the growth of the stroma by the newly formed interlobular and intralobular connective tissue, as indicated by stromal-parenchymal ratio of the organ, as well as reducing of the diameter of the island and the number of endocrine cells in it. β-cells of the islets of Langerhans of children of M group had evidences of functional inhibition manifesting in reducing of the cellular diameter and increase of nuclear/cytoplasmic ratio, decrease in the intensity of ß-cells luminescence. Conclusion. Immunohistochemical study of pancreatic stillbirths from HIV-infected mothers showed a decrease in the intensity of luminescence of ß-cells in sections treated with mAb to insulin, which was the strong evidence of their functional inhibition.

THE STRUCTURAL AND FUNCTIONAL STATUS OF HEART IN CHRONIC HEART FAILURE IN THE PRELIMINARY USE OF PROBIOTICS

Objective: to study the effect of probiotics on the structural and functional state of the heart in stimulated chronic heart failure in rats. Materials and methods. The experiments were performed on male white rats (n = 38). CHF was simulated by double subcutaneous introduction of isoprenaline hydrochloride (Sigma) at a dose of 80 mg/kg at intervals of 24 hours. For histological examination we seized parts of the left ventricular myocardium, prepared cryostat sections of 10 micron thick and stained them by Mallory's method, followed by morphometry using a light microscope MPV-2 («Leitz», Germany) with the digital camera «Leica». The concentration of CRP in serum was measured by agglutination test with latex. Results. It was ascertained that taking the drugs «Dialakt» and «Bifidumbacterin» contribute to preservation of the CMC diameter, which leads to a decrease in fibrosis evidence and areas of undulating deformation and vacuolization of cardiac myocytes (p < 0.05). All the animals with CHF detected certain serum CRP levels (average 172.8 ± 19.2 mg /l), whereas none of the control group revealed it, as it was beyond the sensitivity of the method. In the group of those who had taken «Dialakt», the number of samples with the amount of protein (48 mg/l) reduced to one compared to the animals of group I, while in the group taking «Bifidumbacterin» it reduced to five (86.4 ± 28 mg/l) in the same comparison.

Similar cellular migration patterns from niches in intervertebral disc and in knee-joint regions detected by in situ labeling: an experimental study in the New Zealand white rabbit.

IntroductionPotential stem cell niches (SNs) were recently reported in intervertebral discs (IVDs) and knee joints (KJs) in different mammals (located adjacent to the epiphyseal plate; EP). The aim here was to examine further possible cellular migration and migration directions of cells originating from niches possibly involved in regeneration of cartilaginous tissues in the IVD and in the KJ regions in adult mammals.MethodsIn total, 33 rabbits were used in studies A through C.A. IVD cells were sorted; fluorescence-activated cell sorting (FACS) by size (forward scatter; ≤10 μm or >10 μm or GDF5+ cells (anti-GDF5 antibody). Sorted cells, labeled with cell tracer (carboxyfluorescein-diacetate-succinimidyl ester; CDFA-SE) were applied on IVD explants in vitro. Migrating cells/distance was evaluated by fluorescence- and confocal-microscopy (FC).B. DNA labeling was performed with BrdU (oral administration). Animals were killed (14 to 56 days), KJs collected, and BrdU+ cells visualized with immunohistochemistry (IHC)/anti-BrdU antibody in SN and articular cartilage (AC).C. Cell tracer: (Fe-nanoparticles: Endorem) were injected into SNs of IVDs (LI-LV) and KJs (tibia). Animals were killed after 2 to 6 weeks. Fe-labeled cells were traced by ferric-iron staining (Prussian blue reaction; Mallory method).ResultsA. GDF5+ cells and ≤10-μm cells displayed the best migration capability in IVD explants. GDF5+ cells were detected at a tissue depth of 1,300 μm (16 days). B. BrdU+ cells were observed in early time points in niches of KJs, and at later time points in AC, indicating a gradual migration of cells. C. Fe+ cells were detected in IVDs; in annulus fibrosus (AF) in 11 of 12 animals and in nucleus pulposus (NP) in two of 12 animals. In AC (tibia), Fe+ cells were detected in six of 12 animals. In the potential migration route (PMR), from niches toward the IVD, Fe+ cells (three of 12 animals) and in PMR toward AC (KJs) (six of 12 animals) were detected.ConclusionsResults indicate similar cellular migration patterns in cartilage regions (IVD and KJs) with migration from stem cell niche areas into the mature cartilaginous tissues of both the KJs and the IVD. These findings of a cellular migration pattern in mature cartilage are of interest from tissue-repair and engineering perspectives.

Stainability and Fine Structure of Intracytoplasmic Inclusion Bodies in the Locus Ceruleus of Mouse

The present study compared intracytoplasmic inclusion bodies in the locus ceruleus (LC) of mouse with protein bodies in the LC of human. Phosphotungstic acid-hematoxylin (PTAH) and the Mallory method were used as the anionic stains. The inclusion bodies and protein bodies, which stained with PTAH and the Mallory method in the brain, contain proteins that appear to belong to the same family. Although both inclusion and protein bodies were formed with the same arginine composition, their distribution in the brain was similar and similar physiological changes by morbidity were observed, and the fine structure of the inclusion bodies and protein bodies appeared to be different. The present findings suggest that mouse inclusion bodies and human protein bodies are different.