Clinico-echographic manifestation of endometritis in cows

Purpose: to study the clinical and echographic manifestation of endometritis.Materials and methods. Clinical and echographic research was carried out on 19 red-and-white cows, in the conditions of the agricultural farm of the Druzhba breeding plant, Pavlovsky district, Voronezh region on the 30-32th day after calving using the Easi-Scan scanner from BCF Technology Ltd, Scotland. Before ultrasound examination to establish the diagnosis, all animals were examined, transrectal palpation of the uterus. Simultaneously with the ultrasound examination, a cytological study of the uterine mucosa was carried out. For this purpose, a probe was constructed from a metal catheter intended for artificial insemination of cows with sequins by the deep cervical method with rectal fixation of the cervix; a cytobrush of the Juno probe was attached to the tip of the instrument. To confirm the diagnosis, a laboratory study of cervical mucus was additionally carried out by the express method according to Whiteside, modified by N. I. Polyantsev and Yu. N. Popov.Results. According to the results of rectal examination and ultrasound diagnostics, the cows were divided into three groups: clinically healthy, animals with clinically pronounced chronic endometritis and with latent endometritis. In the group of healthy animals, individual superficial and vacuolated intermediate cells (6.17 ± 0.51) and single neutrophils (2.31 ± 0.32) were detected in smears. In the group of clinically healthy cows, no gram-positive coccobacillary microorganisms and cells of the basal layer of the uterine mucosa were detected. In the second group, in animals with clinically pronounced chronic endometritis, a large number of segmented neutrophils, lymphocytes and a large number of cocci were detected in smears. Thus, in one visual field, the number of gram-positive cocobacillary microorganisms fluctuated on average 764.45 ± 4.56 microbial bodies, while the number of neutrophils increased in comparison with clinically healthy animals by 20.84 times and averaged 48.14 ± 2 in the group. , 91, the percentage of the score averaged around 12.1%. Simultaneously with an increase in the number of leukocytes in animals, the number of epithelial cells of the uterine mucosa increased by 2.21 times, while individual basal cells were detected simultaneously with superficial and intermediate cells (0.75 ± 0.48). In smears obtained from animals of the third group of cows, there was a significant decrease in the number of coccobacillary microorganisms to 75-360 in one visual field in comparison with similar results in the group with a clinically pronounced form of chronic endometritis. The indicator for segmented neutrophils was 8.05 times higher than in clinically healthy cows, while in relation to clinically sick cows by 61.4% and amounted to 18.60 ± 2.23. In terms of the number of superficial, vacuolated intermediate and basal cells in sick animals with chronic clinically pronounced endometritis and latent endometritis, no significant changes were revealed (1.08 times), while in relation to clinically healthy animals, these indicators were 2.38 times higher. The degree of variation in the number of epithelial cells in the group of cows with latent chronic endometritis was insignificant, which indicates the stability of the trait.Conclusion. Ultrasound scanning allows you to identify the latent form of endometritis and establish degenerative changes in the tissues of the uterus. Ultrasound examination in cows in establishing a diagnosis - endometritis should be the decisive diagnostic method.

The Immune Barrier of Porcine Uterine Mucosa Differs Dramatically at Proliferative and Secretory Phases and Could Be Positively Modulated by Colonizing Microbiota

Endometrial immune response is highly associated with the homeostatic balance of the uterus and embryo development; however, the underlying molecular regulatory mechanisms are not fully elucidated. Herein, the porcine endometrium showed significant variation in mucosal immunity in proliferative and secretory phases by single-cell RNA sequencing. The loose arrangement and high motility of the uterine epithelium in the proliferative phase gave opportunities for epithelial cells and dendritic cells to cross talk with colonizing microbial community, guiding lymphocyte migration into the mucosal and glandular epithelium. The migrating lymphocytes were primarily NK and CD8+ T cells, which were robustly modulated by the chemokine signaling. In the secretory phase, the significantly strengthened mechanical mucosal barrier and increased immunoglobulin A alleviated the migration of lymphocytes into the epithelium when the neuro-modulation, mineral uptake, and amino acid metabolism were strongly upregulated. The noticeably increased intraepithelial lymphocytes were positively modulated by the bacteria in the uterine cavity. Our findings illustrated that significant mucosal immunity variation in the endometrium in the proliferative and secretory phases was closely related to intraepithelial lymphocyte migration, which could be modulated by the colonizing bacteria after cross talk with epithelial cells with higher expressions of chemokine.

OPTIMIZATION WAYS OF THERAPEUTIC MEASURES IN WOMEN WITH ENDOMETRIAL POLYPS AND INFERTILITY

Іntroduction. Currently, endometrial polyps (EMP)are the most common pathology of the uterine mucosa,detected in miscarriage and infertility. However, thequestion of the relationship between the mechanisms thatregulate proliferation processes and the morphofunctionaland microbiological features of the endometrium, whichare often confirmed by changes in immunohistochemicalparameters and may be important in treatment, remainsopen.The aim is to optimize the results of treatment inpatients with EMP and infertility by using antioxidantsin combination with immunomodulatory therapy andnonsteroidal anti-inflammatory drugs.Materials and methods. We examined 30 healthywomen who hadn’t had any gynecological diseases,abortions or intrauterine interventions in history and hadnot used intrauterine contraception (the control group), and60 women with EMP and infertility (the main group) byusing clinical, microbiological, bacteriological, ultrasound,hysteroscopic pathohistological and laboratory methods,statistical analysis.Results. In order to assess the effectiveness of theproposed treatment, the main group was divided into twosubgroups: the first one included 30 patients who have beenreceiving the proposed treatment and prevention algorithmand the second one with 30 patients receiving traditionaltreatment.After targeted polypectomy with the basal layer of theendometrium at the site of EMP had been provided, thepatients of the first subgroup were prescribed to take 100mg of doxycycline orally twice a day for 2 weeks and 100mg of vitamin E per day for 4 weeks in combination withimmunomodulatory therapy - cycloferon 12.5% 2.0 mlintramuscularly №10 every other day and anti-inflammatorytherapy with the appointment of rectal suppositoriescontaining non-steroidal anti-inflammatory components - 1suppository per night for 10 days.The patients of the second subgroup were prescribedtraditional antibiotic prophylaxis after hysteroscopy: 100mg of doxycycline orally twice a day for 5 days. Also, thepatients of the main group have been receiving 10 mg ofduphaston twice a day from the 11th to the 25th day of MC.The species composition of the vaginal microflora inwomen with EMP is represented mainly by anaerobic flora.Every second patient with EMP (46%; р І-ІІІ = 0,007) in theabsence of clinical symptoms, and along with a moderateor reduced number of lactobacilli, was diagnosed withopportunistic bacteria, uremicoplasma or Candida fungus.Pregnancies occurred in 9 (47%) of the 19 patients in thefirst natural menstrual cycle after polypectomy and proposedtreatment. Six (31%) women became pregnant during thefirst three menstrual cycles. The remaining patients havebeen observed during six months and were recommendedin vitro fertilization due to long-term infertility. Women ofthe main group who did not plan to get pregnant had norecurrence of EMP for two years. 19 (63.3%) patients ofthe second subgroup faced the recurrence of EMP duringtwo years of follow-up. 11 (30.5%) pregnant womengave physiological childbirth; the labor of 3 patients wascomplicated by hypotonic bleeding in the early postpartumperiod; 3 (11.1%) women are currently pregnant. In patientsof the second subgroup, pregnancy occurred in 5 (16.6%)cases only.Conclusions. The use of the developed method of EMPand infertility treatment allows to restore reproductivefunction in more than half of women. Thus, theimplementation of the method of EMP treatment indicatesits positive effect. In addition, it helps to achieve lastingremission and solves the medical and social problems ofwomen's health and motherhood.

Influence of the diet's structure on the occurrence of the main gynecological pathologies in the sakhalin population of holstein cows

It was established that the level of metabolic energy and the content of raw fiber in diets significantly affects the milk productivity and the occurrence of reproductive pathologies of cows. With a high level of crude fiber and a lack of metabolic energy in the diet, 12.5 % of cows in the herd were diagnosed with persistent ovarian yellow body. Yellow bodies in the ovaries, as shown by surveys, were most often found 25-30 days after delivery or within the same period of time after infertile insemination, as well as in more distant terms. 23 % of sick cows have a diagnosis "ovarian hypofunction", with impaired growth, development, maturation and timely bursting of the follicles. In some cases, ovarian hypofunction occurred as a result of abnormal labor, untimely start-up and physiological old age of cows. In addition to ovarian pathology, cases of pathological processes in the uterus also led to infertility. Endometritis, as a rule, occurred with inflammation of the uterine mucosa (endometrium). This pathology usually occurred in cases of infection, injuries of the uterine mucosa during pathological childbirth, and retention of the afterbirth. With a low level of raw fiber in the diet and a high content of metabolic energy, the most common cows' pathologies were gynecological diseases in 28 heads (23.3 %). Pathological processes in the ovaries with a high level of metabolic energy in the diet almost completely disappear. Most often, endometritis was diagnosed in sick animals - 98.4 %, associated with infection of the genitals and birth injuries. Studies have shown that blood metabolites, body reserves, and the effectiveness of feeding dairy cows depend on the pH of the environment that occurs when feeding different types of diets. One of the conditions for active fermentation of the substrate is the optimal amount of raw fiber. Pathological processes in the ovaries with a high level of metabolic energy in the diet almost completely disappear.

Uterus-specific transcriptional regulation underlies eggshell pigment production in Japanese quail

The precursor of heme, protoporphyrin IX (PPIX), accumulates abundantly in the uterus of birds, such as Japanese quail, Coturnix japonica, resulting in brown-speckled eggshells. The molecular basis of PPIX production in the uterus remains largely unknown. Here, we investigated the cause of low PPIX production in a classical Japanese quail mutant exhibiting white eggshells by comparing its gene expression in the uterus with that of the wild type using transcriptome analysis and performed genetic linkage mapping to identify the causative genomic region of the white eggshell phenotype. We showed that 11 genes, including the 5-aminolevulinic acid synthase 1 (ALAS1) and ferroxidase hephaestin-like 1 (HEPHL1) genes, were specifically upregulated in the wild-type uterus and downregulated in the mutant. We mapped the 172 kb candidate genomic region on chromosome 6, which contains several genes, including a part of the paired-like homeodomain 3 (PITX3), which encodes a transcription factor. ALAS1, HEPHL1, and PITX3 were expressed in the apical cells of the luminal epithelium and lamina propria cells of the uterine mucosa of the wild-type quail, and their expression was downregulated in these cells of the mutant quail. Biochemical analysis using uterine homogenates indicated that the restricted availability of 5-aminolevulinic acid is the main cause of low PPIX production. These results suggest that uterus-specific transcriptional regulation of heme-biosynthesis-related genes is an evolutionarily acquired mechanism of eggshell pigment production in Japanese quail.

Modelling the impact of decidual senescence on embryo implantation in human endometrial assembloids

Decidual remodelling of midluteal endometrium leads to a short implantation window after which the uterine mucosa either breaks down or is transformed into a robust matrix that accommodates the placenta throughout pregnancy. To gain insights into the underlying mechanisms, we established and characterised endometrial assembloids, consisting of gland-like organoids and primary stromal cells. Single-cell transcriptomics revealed that decidualized assembloids closely resemble midluteal endometrium, harbouring differentiated and senescent subpopulations in both glands and stroma. We show that acute senescence in glandular epithelium drives secretion of multiple canonical implantation factors, whereas in the stroma it calibrates the emergence of anti-inflammatory decidual cells and pro-inflammatory senescent decidual cells. Pharmacological inhibition of stress responses in pre-decidual cells accelerated decidualization by eliminating the emergence of senescent decidual cells. In co-culture experiments, accelerated decidualization resulted in entrapment of collapsed human blastocysts in a robust, static decidual matrix. By contrast, the presence of senescent decidual cells created a dynamic implantation environment, enabling embryo expansion and attachment, although their persistence led to gradual disintegration of assembloids. Our findings suggest that decidual senescence controls endometrial fate decisions at implantation and highlight how endometrial assembloids may accelerate the discovery of new treatments to prevent reproductive failure.

Biology and pathology of the uterine microenvironment and its natural killer cells

Tissues are the new frontier of discoveries in immunology. Cells of the immune system are an integral part of tissue physiology and immunity. Determining how immune cells inhabit, housekeep, and defend gut, lung, brain, liver, uterus, and other organs helps revealing the intimate details of tissue physiology and may offer new therapeutic targets to treat pathologies. The uterine microenvironment modulates the development and function of innate lymphoid cells [ILC, largely represented by natural killer (NK) cells], macrophages, T cells, and dendritic cells. These immune cells, in turn, contribute to tissue homeostasis. Regulated by ovarian hormones, the human uterine mucosa (endometrium) undergoes ~400 monthly cycles of breakdown and regeneration from menarche to menopause, with its fibroblasts, glands, blood vessels, and immune cells remodeling the tissue into the transient decidua. Even more transformative changes occur upon blastocyst implantation. Before the placenta is formed, the endometrial glands feed the embryo by histiotrophic nutrition while the uterine spiral arteries are stripped of their endothelial layer and smooth muscle actin. This arterial remodeling is carried out by invading fetal trophoblast and maternal immune cells, chiefly uterine NK (uNK) cells, which also assist fetal growth. The transformed arteries no longer respond to maternal stimuli and meet the increasing demands of the growing fetus. This review focuses on how the everchanging uterine microenvironment affects uNK cells and how uNK cells regulate homeostasis of the decidua, placenta development, and fetal growth. Determining these pathways will help understand the causes of major pregnancy complications.

Menstrual changes in uterine mucosa

Subjecting a systematic study of menstrual blood, Sekiba (Arch, f. Gyn., Bd. 121, H. 1) found that on the 1st day of menstruation, it usually contains already macroscopically distinguishable scraps of sequestered uterine mucose; less often they can be noticed on the 2nd day of regulation, and almost never, on subsequent days.

Proteinase Activated Receptors Mediate the Trypsin-Induced Ca2 + Signaling in Human Uterine Epithelial Cells

Embryo implantation is a complex and tightly regulated process. In humans, uterine luminal epithelium functions as a biosensor gauging the embryo quality and transmitting this information to the underlying endometrial stromal cells. This quality control ensures that only high quality embryos are implanted, while aberrant ones are rejected. The mechanisms of the embryo-uterine mucosa crosstalk remain incompletely understood. Trypsin, a serine protease secreted by the blastocyst, has been implicated in the cross-signaling. Here we address the mechanisms by which trypsin triggers the intracellular calcium signaling in uterine epithelium. We found that protease-activated G-protein coupled receptors are the main mechanism mediating the effects of trypsin in human uterine epithelium. In addition, trypsin activates the epithelial sodium channels thus increasing the intracellular Na+ concentration and promoting Ca2+ entry on the reverse mode of the sodium/calcium exchanger.

Disrupted PGR-B and ESR1 signaling underlies preconceptional defective decidualization linked to severe preeclampsia

Decidualization of the uterine mucosa drives the maternal adaptation to invasion by the placenta. Appropriate depth of placental invasion is needed to support a healthy pregnancy; shallow invasion is associated with the development of severe preeclampsia (sPE). Maternal contribution to sPE through failed decidualization is an important determinant of placental phenotype. However, the molecular mechanism underlaying the in vivo defect linking decidualization to sPE is unknown. Here, we discover the footprint encoding this decidualization defect comprising of 166 genes using global gene expression profiling in decidua from women who developed sPE in a previous pregnancy. This signature allowed us to effectively segregate samples into sPE and control groups. Estrogen receptor 1 (ESR1) and progesterone receptor B (PGR-B) were found highly interconnected with the dynamic network of defective decidualization fingerprint. ESR1 and PGR-B gene expression and protein abundance were remarkably disrupted in sPE. Thus, the transcriptomic signature of impaired decidualization implicates dysregulated hormonal signaling in the decidual endometria in women who developed sPE. These findings reveal a potential footprint that may be leverage for a preconception or early prenatal screening of sPE risk, thus improving prevention and early treatments.