The effect of melatonin on the activity of nucleolar organizer districts in women with ovarian endometriosis

Aim. To study the activity of ribosomal synthesis in women with ovarian endometriosis and its relationship with melatonin levels. Materials and methods. The study included 52 women with ovarian endometriosis. All patients were admitted to the gynecological department of the Clinic of the Southern State Medical University of the Russian Federation for surgical treatment. The paper uses the classification of ovarian endometriosis L. V. Adamyan (1998). During the study, 3 groups were formed: 1 group-stage 1 (n = 8), 2 group – stage 2 (n = 29), 3 group- stage 3 (n = 15).There were no patients with stage 4 ovarian endometriosis. Anamnesis was collected in all women, therapeutic and diagnostic laparoscopy, 6-SOMT was determined in the morning portion of urine, the activity of nucleolar organizer areas in the tissue of endometrioid foci of the ovaries was determined. Statistical data processing was performed using the IBM SPSS Statistics 19 software package. Results. In women with ovarian endometriosis, the lowest level of melatonin in the urine is observed at 1 and 3 stages. In patients with 1 stage ovarian endometriosis, its level was 41.14 ± 1.80 ng / ml; in group 2 – 71.16 ± 2.19 ng/ml, in group 3 – 54.01 ± 1.33 ng / ml (p = 0.048, p1-2 < 0.001, p2-3 = 0.046, p1-3 = 0.062). The main complaint in patients with stage 1 endometriosis was infertility – 8 (100%). The level of melatonin in the urine was significantly reduced in all groups of patients with ovarian endometriosis and infertility, independently of the stage. In patients with 1 stage ovarian endometriosis with low melatonin levels, proliferative diseases of the uterus (uterine fibroids, endometrial polyps, endometrial hyperplasia) are more common in the anamnesis. The number of intranuclear and extranuclear argentaffin inclusions, as well as their combination (INC + ENC) was significantly higher in stage 3 of ovarian endometriosis than in 1 and 2 stages. In endometriosis of the ovaries, a decrease in the level of melatonin in the urine correlates with an increase in the number of cells with a high degree of ribosomal synthesis activity. Conclusion. In women with endometriosis, the lowest values of melatonin were found at stage 1, especially in the presence of infertility, as well as in the common process. The activity of nucleolar organizers, which characterizes the intensity of ribosomal synthesis and the severity of proliferation, was significantly increased in the epithelial cells of ovarian endometriomas at stage 3, while an inverse relationship was revealed between the level of melatonin and the activity of nucleolar organizers.

PSIII-1 Activity of nucleolar organizers depending on genotype

The level of protein proliferation and biosynthesis in various physiological and pathological processes depends on the functional activity in the nucleolar apparatus of cell. The activity degree of the nucleolar organizers varies depending on the level of functional load on the cells. It is associated with the exogenous and endogenous factors and the manifestation of polygenic traits, such as the synthesis intensity of subunits rRNA (18S and 28S) that enter the ribosome. The aim of this research was to study the relationship between the activity of nucleolar organizers and the animal genotype. The objects of study were goats with different genotypes divided by 3 groups of goats hybrids were used: group_I was 5 individuals of Karachay × F1(C.sibirica×C.hircus); group_II - 4 individuals of Boer × F1; group_III - 3 individuals of F2(C.sibirica×C.hircus) × F1. The material for the study was venous blood. Blood smears were fixed with Lillie’s solution and were stained using the Havel and Blake technique. Microscopic studies were performed using a Nikon Eclipse Ni equipped with Nikon DS-Qi2 camera (4908×3264). For statistical analysis of the obtained data, the SPSSv.23 was used. We used the average optical density AgNOR (DNOR) as an indicator reflecting the total amount of Ag bound to the argyrophilic proteins of the nucleolar organizer, It was found that in goats of the second group this indicator was 158.58±10.18, which was significantly inferior in terms of AgNOR to animals in group_I (211.10±3.62) and in group_III (206.77±4.43). No significant differences between the goats of groups I and III were found. Thus, the AgNOR criterion depends on the goat genotype to a certain extent and is of interest for assessing the activity of the nucleolar organizers in goats. Supported by RFBR project №20-016-00116А and the Ministry of Science and Higher Education of Russia (№АААА-А18-118021590132-9).

Activity of nucleolar organizers in endometrial glandular and surface epithelial cells in patients with external genital endometriosis

Hypothesis/aims of study. Endometriosis is a disease characterized by proliferation of tissue that is similar in morphology and function to the endometrium, outside the uterine cavity. Thanks to the theory of J. Sampson (1927), which explains endometrioid heterotopia formation by bits of endometrial tissue migrating through the fallopian tubes during menstruation and their subsequent implantation in the pelvis, research has begun on the biological properties of eutopic endometrial cells in women with endometriosis, which is still relevant today. The present study was aimed to assess the activity of nucleolus organizers in the endometrium in women with external genital endometriosis and without it at different stages of the menstrual cycle. Study design, materials, and methods. Paraffin blocks of endometrial biopsy specimens were taken from 68 women with and without external genital endometriosis. The activity of nucleolar organizers was evaluated by the method of J. Crocker (1983). Results. The data obtained showed a significant increase in ribosomal activity in the cells of the endometrial glandular and surface epithelium in women with external genital endometriosis at the stages of late proliferation, as well as early and middle secretions. There was also a significant decrease in the activity of the nucleolar organizers of endometrial epithelial cells at the stage of late secretion in women with external genital endometriosis. Conclusion. The obtained differences in regulation of ribosomal synthesis in the cells of the integument and glandular epithelium of the endometrium in women with external genital endometriosis, in our opinion, can provide an increase in the total amount of epitheliocytes of the endometrium, increase their adhesive and invasive abilities, and lengthen the life cycle of cells. This contributes to the formation of endometrioid heterotopies and the development of external genital endometriosis. Changes in the activity of nucleolar organizers in the endometrium in women with external genital endometriosis may be of clinical interest for the diagnosis of this pathology.

Genome instability in Chironomus annularius sensu Strenzke (Diptera, Chironomidae): A biomarker for assessment of the heavy metal contaminants in Poland

We studied the effect of heavy metal contaminations on genome instability of Chironomus annularius sensu Strenzke from three sampling sites on the River Chechło valley in southern Poland and a control site in Saraya, Bulgaria. The analysis of the heavy metal concentrations in sediments at the three river sites indicated concentrations higher than those in the fossil reference data (Zn 83-160 times, Pb 64-250 times, and Cd 210-793). According to the geoaccumulation index (Igeo), the sediments at all Chechło stations were classified as extremely polluted (Igeo class 6) by Cd, Pb and Zn while they were much less polluted by Cu (classes 1-3). The concentration of Cd, Pb and Zn in the sediments studied exceeded 21-79, 11-44 and 18-34 times the toxic effect threshold (TET) respectively; therefore, the sediments potentially have detrimental effects on the fauna. Genome instability was determined by somatic structural chromosome alterations (mainly heterozygous inversions) in the salivary gland chromosomes of C. annularius. On the basis of the number and frequency of these aberrations the somatic (S) and cytogenetic (C) indices were calculated as these indices allow a better quantification of the impact of heavy metals on organisms living in sediments. In the three Chechło sampling sites somatic aberrations occurred at a frequency significantly higher than in the control site (P<0.001). The three Chechło samples possess a high spectrum of somatic rearrangements with S index varying from 1.5 to 2.23 while in control site, S is 1.08. The C index of C. annularius from the polluted river sites varied from 0.163 to 0.380, far in excess of the C value of the control site (1.0), indicating strong pollution (<0.5) according to previous studies. In addition, a high level of inherited aberrations was detected (19 inherited band sequences, two of which were new). In all river stations the inverted homozygous band sequences A2.2 occurred in a high frequency (85%) and no standard sequences (A1.1) were observed. The transcription activity of Nucleolar Organizers (NORs) in chromosome arms A, C, E, G was often damaged and NOR activity was completely suppressed in 12.48% of larvae. Moreover, appearance of malformations in the external morphology of larvae was detected in 16.67% of larvae from all polluted sites. Therefore, the genome instability as estimated by two cytogenetic indices is a good biomarker of long-term toxicity and could be successfully implemented in the sediment monitoring procedures.

Analysis of the karyotype of Callisia elegans Alexand. (Commelinaceae) including differential staining of chromosomes

The number and morphology of <em>Callisia elegans</em> Alexand. chromosomes were studied employing staining with acetic carmine and differential Giemsa staining. It was found that its karyotype was 2n = 12 chromosomes, whose lengths fell in the range of 16.8 to 8.8 µm. The chomosomes, arranged in order of length, were classified respectively to types: sm, t, t, t, t, st. The distribution of C-banding is given for this karyotype. The presence of microsatellites on the long and short arms was found in the chromosomes of the second pair. Frequently there were 4 nucleoli of unequal size in interphase nuclei. In many cells, lower numbers of nucleoli (3-1) were seen which was -probably due to their fusion. The maximum number of nucleoli corresponded to the number of nucleolar organizers accompanying the satellites.