Uterine Horn Research Articles

Investigation of the Role of Pituitary Adenylate Cyclase-Activating Peptide (PACAP) and Its Type 1 (PAC1) Receptor in Uterine Contractility during Endometritis in Pigs.

Uterine inflammation is a common pathology in animals, leading to disturbances in reproductive processes and reduced production profitability. Pituitary adenylate cyclase-activating peptide (PACAP) effects at the uterine level during inflammation are not known. In the current study, we analyzed the relative PACAP type 1 receptor (PAC1R) mRNA transcript and protein abundances in the myometrium (MYO), as well s PACAP and PAC1R involvement in the contractile function of inflamed pig uterus. To that end, E. coli suspension (E. coli group) or saline (SAL group) was injected into the uterine horns or laparotomy was performed (CON group). Eight days after the bacteria injections, severe acute endometritis and a reduced relative abundance of PAC1R protein in the MYO were observed. Compared to the period before PACAP in vitro administration, PACAP (10−7 M) in the CON and SAL groups decreased in amplitude in the MYO and endometrium (ENDO)/MYO, whereas in the E. coli group, increased amplitude in the MYO and reduced amplitude in the ENDO/MYO were observed. In the E. coli group, PACAP enhanced the amplitude in the MYO (10−7 M) and decreased the amplitude in the ENDO/MYO (10−8 M) compared with other groups. PACAP (10−7 M) increased the frequency of both kinds of strips in the CON and SAL groups compared with the pretreatment period. PACAP (both doses) did not significantly change the frequency in the E. coli group, whereas in response to PACAP (10−7 M), the frequency was reduced compared to other groups. In the MYO, PAC1R antagonist decreased the amplitude reduction (CON and SAL groups) and reversed a rise in PACAP (10−7 M)-evoked amplitude (E. coli group). PAC1R blocking reversed (MYO) and abolished (ENDO/MYO) the stimulatory effect of PACAP (10−7 M) on the frequency (CON and SAL groups). PAC1R antagonist and PACAP (10−7 M) evoked the appearance of frequency depression in both kinds of strips (E. coli group). In summary, in pigs, severe acute endometritis reduces the relative abundance of PAC1R protein in the MYO, and PAC1R mediates the influence of PACAP on inflamed uterus contractility.

Clinico-haemato-biochemical and Pathological Alteration of Pyometra in Canines

Background: Pyometra is a pathological state of the compromised uterus of grown-up to old, diestrum bitches. Pyometra is easy to identify but can be more challenging when there is no vaginal discharge. In the present study, an attempt was made to find out the pathogen involvement and clinico-haemato-biochemical alteration in bitches affected with Pyometra. Methods: Twelve bitches with a history of inappetence, polydipsia, dirty vaginal discharge, occasional vomition irrespective of breeds and age groups were clinically examined and diagnosed as pyometra. The presumptive clinical diagnosis was based on case history, clinical signs, vaginal cytology, abdominal ultrasonography or radiography, gross examination of a pus-filled uterus after the ovariohysterectomy. Result: Vaginal examination revealed brown chocolate malodorous vaginal discharge with discolouration of the perivulval tissue. Abdominal ultrasound showed hyperplasia with echogenic material in the uterine wall along with distended uterine horns containing a large volume of cellular contents. Radiographic imaging revealed cranio-dorsal displacement of the small intestine and a twisted uniform tubular opacity with a distended uterus. The vaginal cytology revealed a large number of degenerated neutrophils, intermediate and parabasal cells. Escherichia coli were the major pathogen isolated from the pyometra uteri of bitches. The surface of the endometrium was columnar ciliated epithelium, with hyperplasia of subepithelial tissue and thick muscularis. The haemato-biochemical evaluation in pyometra infected bitches revealed decreased haemoglobin (Hb), packed cell volume (PCV), total erythrocytic count (TEC) and lymphocyte count indicating the microcytic hypochromic type of anaemia. Leucocytosis with a predominant absolute neutrophilia and lymphocytosis were consistently found in canine pyometra. There was a significantly elevated profile of blood urea nitrogen, creatinine, transaminases and alkaline phosphatase as well as total proteins observed in pyometric female dogs as compared to healthy ones which may have diagnostic and prognostic importance, which also indicated damage of vital organ caused by bacterial endotoxins. The haemato-biochemical indices correlating with imaging techniques, vaginal cytology and histopathological examination can serve as good prognostic markers in bitches with pyometra.

Technical notes surgical treatment of a ruptured interstitial pregnancy

Non-tubal ectopic pregnancies represent fewer than 10% of all ectopic pregnancies. However, they are associated with a high rate of mortality due to late diagnosis and uterine horn rupture which requires radical emergency surgical management. Cornuectomy via laparoscopy is a treatment of choice. We provide here a simple description of laparoscopic cornuectomy using an Endo GIA stapling system: the Endo GIA® automatic forceps. It has the advantage of removing the mass, suturing, and achieving satisfactory haemostasis in a single step.

TGF-β signaling is important for female reproductive tract resident memory CD8 T cell development

Abstract Tissue-resident memory T cells (TRM) patrol non-lymphoid barrier tissues and provide rapid protection against reinvading pathogens. While TRM formation in several barrier mucosal tissues including intestine and respiratory tract has been investigated, factors that control TRM establishment in the female reproductive tract (FRT) remain unclear. Here we established a model of local FRT infection, characterized CD8 TRM development and probed the role of cytokine TGF-β in TRM formation. We adopted a mouse model of intravaginal lymphocytic choriomeningitis virus (LCMV) infection that generated robust local CD8 T cell response in both the lower FRT (cervix, vagina) and upper FRT (ovaries and uterine horn). Upon viral clearance, CD8 T cells in the tissue gradually adopted mature TRM phenotype. Parabiosis confirmed most antiviral T cells in the FRT were truly resident. There were marked differences in the phenotypes of TRM in upper and lower FRT with CD103+ epithelial TRM predominating the lower FRT mucosa. Longitudinal analysis revealed TRM in both compartments undergoes attrition. The cytokine TGF-β has been implicated in TRM development in number of barrier and visceral organs. Using TGF-β RII deficient transgenic CD8 T cells, we observed that lack of TGF-β receptor signaling impairs acquisition of CD103 and CD69 expression by FRT TRM. However, TGF-β RII deficient CD8 T cells showed elevated expression of effector molecules granzyme-B and interferon-gamma upon in vivo stimulation. Taken together, our data suggest a key role of TGF-β in the formation and function of virus specific TRM in the FRT. Further investigation will provide mechanistic insight on the nature of TGF-β signaling pathway impacting FRT TRM formation. Supported by NIH grants (P20GM121298 and P20GM109035) and Searle Scholars Foundation

Modulatory role of 17α‐estradiol (non‐feminizing estrogen) on 6‐OHDA‐induced Parkinsonism in ovariectomized rats

BackgroundEpidemiological studies have reported that the incidence of Parkinson’s disease (PD) is 1.5 times more in men as compared to women. It suggests that gonadal hormones influence the development and progression of PD. Estrogen replacement therapies have a protective effect against dopaminergic toxins and can be protective in Parkinsonism. However, there are multiple unresolved issues regarding the safety of ERT. Thus, there is a essential need to find alternatives that would avoid these health risks.ObjectivesThus main objective of study was to assess effect of 17α‐estradiol, a non‐feminizing estrogen, in attenuating PD employing an animal model of surgical menopause.Methodology17α‐estradiol and 17β‐estradiol was injected 10 µg/kg/s.c. for 14 days in ovariectomized 6‐OHDA rats. After 14 days of drug administration behavioral parameters of Parkinsonism (sensorimotor activity, motor coordination, and locomotor activity) were performed. These behavioral parameters were further correlated with various biochemical and molecular markers in postmenopausal Parkinsonism. Uterine weight and serum estradiol levels were also assessed to determine estrogenic effect of both the drugs peripherally. Effect of both drugs on expression of ER‐α and ER‐β in striatum was also evaluated. Further the differential effect of 17α‐estradiol and 17β‐estradiol was evaluated in postmenopausal Parkinsonism.Results17α‐estradiol and 17β‐estradiol equally modulated behavioral changes and attenuated rise in oxidative stress markers, inflammatory markers, improving neurotrophic factors and modulation of striatal neuron morphology. 17β‐estradiol treatment restored the uterine weight and serum estradiol levels towards normal, however 17α‐estradiol did not show any effect on uterine horns and serum estradiol levels. No effect of 17α‐estradiol was observed on expression of ER‐α and ER‐β in the striatum, indicates receptor independent effect of 17α‐estradiolConclusionBoth 17α‐estradiol and 17β‐estradiol are equally effective in Parkinsonism, however, 17α‐estradiol may prove to be safer alternative of 17β‐estradiol in Parkinsonism due to lack of feminizing effect.

Oxytocin, Prostaglandin F2α, and Prostaglandin E2 Induced Contractions in Isolated Uterine Smooth Muscle Strips from Non‐pregnant, Late Pregnant, and Postpartum Rats

Objective/IntroductionNumerous studies have investigated uterotonic agents (e.g., oxytocin) in uterine smooth muscle (USM) from non‐pregnant (NP) and late‐pregnant (LP) women and/or animal models of pregnancy. Surprisingly, few studies have investigated USM from the post‐partum period to determine whether USM contractility returns to its pre‐pregnant state. We designed a study in rodents to investigate whether USM contractility to oxytocin (OXY), prostaglandin F2α (PGF2α), and prostaglandin E2 (PGE2) returned to pre‐pregnant values within 30 days postpartum.MethodsStudies were performed in NP (n=12), LP (21d gestation, n=4), 3 days post‐partum (PP3, n=2), and 30 days post‐partum (PP30, n=4) Sprague Dawley rats. Uterine horns were cleaned of connective tissue and cut along the longitudinal axis allowing isolation of USM (width: 2mm, length: 4±1mm) oriented in the longitudinal muscle (LM) or circular muscle (CM) direction. CM or LM strips were mounted in tissue baths containing Krebs and equilibrated @ 37°C for 60 min prior to starting one of the two following protocols.Protocol 1 identified the optimal resting tension (in grams: 0.25, 0.50, 1.0, and 1.5) to measure spontaneous contraction (SCA), K+‐induced contraction (60mM K+), and cumulative OXY‐induced contraction (10‐11 to 10‐7) in CM and LM of NP rats (N=4‐6).Protocol 2 was designed assess SCA, K+‐induced contraction, cumulative concentration‐response relationships to OXY (10‐11 to 10‐7), PGF2α (10‐8 to 10‐4), or PGE2 (10‐8 to 10‐4) in CM and LM, of NP, LP, PP3, and PP30 rats (N=2‐6).Statistical comparisons were made using either a t‐test or two‐way ANOVA, and considered statistically different at P<0.05.ResultsProtocol 1‐ LM SCA was greater than CM SCA at all resting tensions. K+‐induced constrictions in LM and CM were similar at each resting tension. OXY‐induced contraction in LM was generally greater than CM (P<0.05 at 0.25g, 0.5g, and 1.5g tension, comparable at 0.5, 1.0, and 1.5g tension).Protocol 2 ‐ SCA in NP, LM was greater than NP, CM (P<0.05). There was no difference in SCA in CM vs LM in USM from LP, PP3, and PP30 animals. K+‐induced constrictions in CM and LM were similar in USM from NP, LP, PP3 and PP30 animals. OXY‐induced contractions were greater in LM than CM for all pregnancy states (P<0.05). PGE2 contractions were greater in LM vs CM in USM from NP rats (P<0.05), but were similar in USM from LP, PP3, and PP30 animals. In contrast, PGF2α‐contractions were similar in LM and CM for all pregnancy states.Summary/ConclusionOur preliminary data suggests that uterotonic agents be assessed at either 0.5g or 1.0g resting tension. In addition, uterotonic agents exhibit significantly greater contraction in longitudinal vs circular orientation. Finally, contractility to uterotonic agents returns to pre‐pregnant values by PP30.

Endometrial organoids derived from Mayer-Rokitansky-Küster-Hauser syndrome patients provide insights into disease-causing pathways.

ABSTRACTThe uterus is responsible for the nourishment and mechanical protection of the developing embryo and fetus and is an essential part in mammalian reproduction. Mayer–Rokitansky–Küster–Hauser (MRKH) syndrome is characterized by agenesis of the uterus and upper part of the vagina in females with normal ovarian function. Although heavily studied, the cause of the disease is still enigmatic. Current research in the field of MRKH mainly focuses on DNA-sequencing efforts and, so far, has been unable to decipher the nature and heterogeneity of the disease, thereby holding back scientific and clinical progress. Here, we developed long-term expandable organoid cultures from endometrium found in uterine rudiment horns of MRKH patients. Phenotypically, they share great similarity with healthy control organoids and are surprisingly fully hormone responsive. Transcriptome analyses, however, identified an array of dysregulated genes that point to potentially disease-causing pathways altered during the development of the female reproductive tract. We consider the endometrial organoid cultures to be a powerful research tool that promise to enable an array of studies into the pathogenic origins of MRKH syndrome and possible treatment opportunities to improve patient quality of life.

Unicornuate Uterus with Noncommunicating Rudimentary Horn (Class U4aC0V0/ESHRE/ESGE Classification) and a Communicating Bladder Endometriotic Nodule

To describe the diagnostic workup and laparoscopic management of a noncommunicating left uterine rudimentary horn (class U4aC0V0 European Society of Human Reproduction and Embryology/European Society of Gastrointestinal Endoscopy Classification) with communicating endometriotic bladder nodule. Step-by-step description of the surgical treatment. A 33-year-old woman with unicornuate uterus and a left-side noncommunicating rudimentary horn affected by primary infertility, mild dysmenorrhea (visual analog scale score 6), severe catamenial dysuria (visual analog scale score 10), and catamenial hematuria. Noncommunicating rudimentary horns are rare Müllerian anomalies present in 20% to 25% of women with a unicornuate uterus. It is associated with severe dysmenorrhea, pelvic pain, subfertility, and poor obstetric outcomes and usually presents with cyclic pelvic pain that starts early after the menarche. Endometriotic bladder nodules are present in 1% to 2% of patients with endometriosis. In the literature, there are no reported cases of noncommunicating rudimentary horn with communicating endometriotic bladder nodules. Surgical excision of the rudimentary horn is the treatment of choice. In our case, the 2-dimensional/3-dimensional ultrasound revealed a right unicornuate uterus with a left noncommunicating rudimentary horn with hematometra. The uterine fundus presented «gamma sign» vascularization. In addition, a bladder endometriotic nodule (16×15 mm) communicating with hematometra was displayed. Magnetic resonance imaging demonstrated no additional malformations. Diagnostic hysteroscopy revealed a single cervix without vaginal malformation and small right uterine cavity with single tubal ostium. At laparoscopy, using hysteroscopic transillumination, a clear plane of dissection was identified between the rudimentary horn and the uterus confirming the presence of a noncommunicating horn. Evaluation of the abdominal cavity showed bilateral normal adnexa with normal ovaries. Chromopertubation showed a patent right Fallopian tube and obstructed left tube. A left salpingectomy using bipolar and the ultrasonic energy was performed. The utero-ovarian ligament was transected, and the left ovary was preserved. The left ovary was suspended at the pelvic wall, the retroperitoneum was opened, the ureter was identified, and the left uterine artery was temporary occluded. The left round ligament was transected and the left paravesical space was developed. With a lateromedial approach, we opened the vesicouterine septum to dissect the bladder from the rudimentary horn. The endometriotic bladder nodule was gently detached from the uterine horn with a lateromedial approach. The left uterine artery was coagulated and dissected at level of the cervix. A solution of vasopressin was injected between the uterine horn and the uterus. Resection of the rudimentary horn was performed. The peritoneum was closed. The temporary occlusion of the uterine artery was removed. The specimen was placed in a bag and removed using an extracorporeal tissue extraction technique. The late clinical presentation of our patient with only mild dysmenorrhea could be explained by the drainage of the hematometra from the noncommunicating horn into the endometriotic bladder nodule. The bladder symptoms in patients with Müllerian anomalies should be carefully investigated. The laparoscopic removal of rudimentary horn with mobilization of communicating bladder nodule when present is a safe and feasible method to improve symptomatology.

Rudimentary Corneal Rupture of A Unicornuate Uterus in A 23-Week Pregnancy: About A Case

The rudimentary uterine horn belongs to the group of major uterine malformations according to the classification of Musset's classification. This rudimentary horn may have a cavity with endometrial lining of the uterus, and thus can serve as a site of implantation during pregnancy. The presence of a pregnancy in a rudimentary horn is a rare event, which can be revealed by a uterine rupture. It leads to an emergency fetal extraction and it is then recommended to perform an exeresis of this rudimentary horn [6, 7]. If it is not performed, it theoretically exposes the risk of recurrence of uterine rupture in case of a new pregnancy in this rudimentary horn. There is a need to increase awareness of this disease, especially in developing countries where the possibility of detection of the disease is limited. We report a case of ruptured rudimentary horn pregnancy in shock at 23 weeks of gestation misdiagnosed as intrauterine pregnancy. The pregnancy was located in a rudimentary left uterine horn ruptured on its posterior surface and communicating with the right horn. A hemi hysterectomy was then performed with the removal of the rudimentary horn and the homolateral adnexa. The fetus was deadborn. The postoperative course was simple and the patient was discharged on the fifth postoperative day.

Changes in selected cytokines, acute-phase proteins, gonadal hormones and reproductive organs of non-pregnant does challenged with Mannheimia haemolytica serotype A2 and its LPS endotoxin.

Previous investigations have revealed that lipopolysaccharide (LPS) endotoxin from certain Gram-negative bacteria could adversely affect the reproductive system of female animals. However, it is unknown whether LPS endotoxin of Mannheimia haemolytica serotype A2, the principal causative bacteria that cause pneumonic mannheimiosis in small ruminants, may also induce similar insidious effects. Therefore, this study aimed to investigate the effects of M. haemolytica serotype A2 and its LPS endotoxin on the responses of female gonadal hormones (progesterone and oestrogen), pro-inflammatory cytokines (interleukin-1β, interleukin-6), acute-phase proteins (haptoglobin and serum amyloid A) and cellular changes via histopathology study of female reproductive organs of the treatment does. Twelve clinically healthy, non-pregnant, crossbred does were randomly allocated into three equal groups. Group 1 was administered intranasally with 2ml of sterile phosphate-buffered saline (PBS) and served as a negative control group. Group 2 was challenged intranasally with 2ml of bacterial inoculum containing 109 colony-forming units (CFU)/ml of M. haemolytica serotype A2, while Group 3 was challenged intravenously with 2ml of LPS endotoxin extracted from 109CFU/ml of M. haemolytica serotype A2. Following that, blood samples were collected serially at pre-determined intervals for serological analyses. All does were euthanised 60days post-challenges, and tissue samples from the ovaries, oviducts, uterine horns, uterine body, cervix and vagina were collected for histopathological study. The serological result revealed a significant increase (p < 0.05) in the mean concentrations of progesterone, oestrogen, interleukin-1β, interleukin-6, haptoglobin and serum amyloid A for both challenged groups. Histopathologically, all reproductive organs (except the cervix and vagina) from both challenged groups displayed significant cellular alterations (p < 0.05) characterised by haemorrhage and congestion, necrosis and degeneration, inflammatory cell infiltration and oedema. This study provides new information that elucidates the potential role of pneumonic mannheimiosis in the pathogenesis of female infertility amongst small ruminants.

Monitoring Influence of PGF2 , Estrogen and Oxytocin on α Postpartum Uterine Involution by Ultrasonography in Ewes

The goal of this study was designed to investigate the effect of estrogen, oxytocin and PGFα on uterine involution post-partum (p.p). In addition, the times for ewes to go through involution by ultrasonographical rectal examination lead to assess the best time for insemination that ensure the highest fertilization rate to avoid endometrit is post-partum. This study carried out at sheep and goat's improvement station, state board of agricultural research in Agargof, during November 15,2020 to April 15, 2021 This study involved 30 ewes of 2-4 years old and . 50-65kg body weight. Blood was collected to estimate progesterone concentration and the role of ovarian activity. Results showed a gradual decrease in uterine lumen and uterine diameter. The uterine horn diameter, at different weeks of post-partum in groups treated PGF2 was α40.63 mm at first week and decrease14.22 mm The uterine involution in ewes treated with PGF2 on post-partum, facilitate uterine involution . α process in ewes and lead to remove uterine fluid (lochia) and decrease uterine size, and uterine lumen. The study proves progesterone concentration as indicator to detect Ewes' post-partum ovarian activity. The current study reveals that the ultra-sonographic approach is a viable option for uterine involution management.

42 Genome-wide Association Study of Ovary Characteristics in Admixed Beef Heifers Through Comparative Bayesian Approaches

Abstract Reproductive performance of heifers is a key indicator of herd productivity. The objective was to evaluate markers and genes associated with reproductive tract characteristics through a comparative approach. For this study, 295 admixed beef heifers were examined via transrectal ultrasonography at approximately 13 months of age. Exploratory factor analysis of antral follicular count (count per size, side, and total), length, height and diameter for left and right sides of ovaries, and uterine horn diameter revealed a latent variable that included total antral follicle count and ovary diameters per side (n = 3 traits), termed OVARY. Confirmatory factor analysis produced OVARY values per heifer. Quality controls left 116,133 SNP per heifer for association analyses. A marker window-based univariate approach with OVARY was conducted using an empirical Bayesian approach adjusting for nested population structure using marker frequency differences (Method 1) as well as an MCMC Bayesian approach identifying the fraction of SNP to be included with population structure as a fixed effect (Method 2). SNP with high genetic variance in important windows (PPAW &amp;gt; 0.75) were mapped to their closest gene for enrichment analysis. Using Method 1, 2 genes were found to be associated and enrichment analysis identified SMYD2 contributing generally to epigenetic and transcriptional regulatory pathways and GO terms (adjusted P &amp;lt; 0.05). Using Method 2, 11 genes were discovered to be associated with OVARY (adjusted P &amp;lt; 0.05). Three of 11 genes were involved in neural transduction, cornification, epithelial integrity, and adipogenic pathways and GO terms. Eight of 11 genes were involved in reproductive hormonal signaling, post-natal growth, sex determination and reproductive organs development, embryo implantation, and signaling transduction GO terms. Given this, Method 2 provides better insights towards the association of markers and genes for future selection of admixed beef heifers with overall better reproductive characteristics than Method 1.

Comparison of pregnancy rate in dromedary camel between early-stage embryos and blastocyst transfer produced by somatic cell nuclear transfer using in vitro-matured oocytes.

We compared the pregnancy and live birth rates following transfer of early-stage embryos or blastocysts produced by somatic cell nuclear transfer using in vitro-matured oocytes. In total 102 ovaries were collected from dromedary camels at a local abattoir; from these 1048 cumulus-oocytes complexes (COCs) were aspirated and cultured for 42 h in a commercial maturation medium. Metaphase II oocytes were subjected to nuclear transfer. Somatic cell nuclear transfer-derived embryos were cultured in a commercial embryo medium for 2 or 7 days. Next, 71 early-stage embryos were surgically transferred to the left fallopian tube of 28 recipients and 47 blastocysts were transferred to the left uterine horn of 26 recipients. Early pregnancy was detected by serum progesterone (P4), and pregnancy was confirmed using ultrasonography on days 30 and 90 after embryo transfer. Pregnancy rate based on P4 level was 17.86% (5/28) and 11.54% (3/26) for early-stage embryo and blastocyst transfer, respectively. In the early-stage embryo group, out of five recipients, one recipient had lost the pregnancy by the first ultrasonography on day 30; two other recipients aborted at 14 and 24 weeks, and two recipients gave live births. In the blastocyst group, out of three recipients, one lost the pregnancy at an early stage and two recipients gave live births. Therefore, for dromedary camels, we recommend transvaginal blastocyst transfer from the standpoint of the pregnancy and live birth rate, ease of the transfer procedure, and comfort and safety of the recipients.

Effect of Human Peripheral Blood Mononuclear Cells on Mouse Endometrial Cell Proliferation: A Potential Therapeutics for Endometrial Regeneration

Objectives: The persistently thin endometrium is a major cause of repeated implantation failure; however, there is no definite treatment for it yet. This study aimed to confirm the potential of human peripheral blood mononuclear cells (hPBMCs) as a therapeutic agent for endometrial regeneration. Design: An experimental study was carried out. Participants/Materials, Setting, Methods: To assess the in vitro effect of hPBMC, the human primary endometrial epithelial cell lines SNU-685 and SNU-1077 were co-cultured with or without 1 × 105 hPBMCs for 24 h. To evaluate the in vivo effect, either 1 × 105 hPBMCs in PBS or PBS alone were injected into the left uterine horn of nonobese diabetic-severe combined immune-deficient mice, and the right untreated uterine horn was used as control. Results: Co-culture with hPBMCs stimulated significant proliferation in both SNU-685 and SNU-1077 cell lines (p = 0.002 and 0.044, respectively). Moreover, treatment with hPBMCs significantly increased the thickness in all parts of the endometrium compared with that in the untreated control uterine horn (proximal: 1.69 ± 0.19 vs. 1.00 ± 0.10, p = 0.009; middle: 1.51 ± 0.14 vs. 1.00 ± 0.12, p = 0.010; distal: 1.72 ± 0.22 vs. 1.00 ± 0.12, p = 0.003, respectively). Compared with the PBS injection group, the hPBMC injection group had significantly thickened endometrium in the middle (p = 0.036) and distal segments (p = 0.002) of the uterine horn. Immunohistochemical analysis revealed the presence of exogenously injected hPBMCs in the uterus of recipient mice. hPBMC-recipient mice had cyclic uterus with normal histology in the endometrium. Limitations: hPBMCs were not applied directly to a mouse model with thin endometrium, so further study is needed. Conclusion: The beneficial effect of hPBMCs on endometrium may suggest their clinical feasibility for the safe treatment of infertile patients with persistently thin endometrium.

Spontaneous embryonic death in plains viscacha (Lagostomus maximus - Rodentia), a species with unique reproductive characteristics

Spontaneous embryonic death is a conserved reproductive event in Eutherians. The macro and microscopic characteristics of this type of death are similar between the different taxa. However, in the hystricomorphic rodent plains viscacha (Lagostomus maximus) is exceptional in terms of massiveness (80% embryonic resorption). In this species, of the 10–12 implantation sites (IS) (half in each uterine horn), only the caudal embryos will survive, resorbing the cranial and intermediate IS. We hypothesize that uterine structural variations in L. maximus restrict growth and promote embryo death, with the consequent loss of placental homeostasis in the cranial and middle IS. In this study, different studies (ultrasonography, macroscopy and microscopy) were carried out to analyze different aspects of the intermediate gestation of L. maximus (46 days postcoitus). Ultrasonographic studies revealed that the cranial and middle IS (IS-1, IS-2, and IS-3) had no recognizable embryonic and placental structures as compared to the caudal implantation sites (IS-4). Macroscopically, the areas corresponding to the embryos in the cranial and middle IS were occupied by a necrotic black semi-fluid mass. Moreover, the placenta in these IS was undifferentiated. However, in the caudal IS both the embryo and its placenta were distinguishable. Using histological and immunohistochemical techniques, it was observed that the placentas of IS-1, IS-2 and IS-3 were disorganized and showed hemorrhage, inflammatory infiltration containing neutrophils, macrophages, mast cells and foreign body giant cells, apoptotic trophoblast, and a layer of collagen fibers and fibroblasts that circumscribed each of these IS. In contrast, the placenta of the caudal IS showed an organized maternal-embryonic interface. The characteristics observed in IS in resorption of viscachas in intermediate gestation show that, regardless of gestation time, embryonic death has a similar macro and microscopic morphological pattern among eutherians with invasive placentation. However, the massiveness and sectorization of embryonic death in the plains viscacha make the species a unique model for the study of this reproductive event.

Plasminogen activator activity and plasminogen activator inhibition in the uterus of ewes after the induction of oestrus synchronization or superovulation, involving eCG

During the preimplantation period, early mammalian embryos largely depend on uterine environment. Concurrently, uterine architecture undergoes substantial growth and remodeling under the influence of ovarian steroids. Plasminogen activators/plasmin proteolytic system that is implicated in tissue remodeling is affected by reproductive hormone variations. Thus, hormone treatments for reproductive function control possibly influence proteolytic enzyme activity in the reproductive tract and its study in uterine fluid and endometrium could demonstrate possible changes in uterine environment. This work examines (a) the effect of hormone treatment involving eCG in the dosage used for oestrus synchronization or for superovulation on plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) in the uterus of ewes before maternal recognition of pregnancy and (b) the possible relationships among blood ovarian steroid concentration, proteolytic enzyme activity and ovarian response/embryo yield. Chios ewes (n = 24) were treated with progestagen intravaginal sponges for 12 days and eCG (500 or 1000 IU) at sponge removal; ewes at the second oestrus after synchronization were used as controls. On day 6 of the oestrous cycle, after slaughter, ovarian response was assessed and embryos were collected; uterine horn flushings (UHF), caruncular endometrium (CE) and intercaruncular (ICE) endometrium samples were collected, for PAA and PAI photometrical assessment. Oestradiol-17β (E2) and progesterone (P4) concentration was assessed in blood serum samples collected before slaughter by a radioimmunoassay. A significant increase of PAA in the UHF was observed after treatment; PAA in UHF was significantly associated with the number of embryos or high-quality embryos, while PAA in UHF per embryo or high-quality embryo was not affected by treatment. No significant effect of treatment was noticed on PAA or PAI in the endometrium (CE, ICE). In UHF and in ICE, PAI against tissue-type PA had a significant positive relationship with PAI against urokinase-type PA. PAI against tissue-type PA in CE had significant negative relationship with E2 concentration. In conclusion, hormone treatment involving eCG in the dosage used for superovulation and, to a lesser degree, in the dosage used for oestrus synchronization affected PAA in the uterus of Chios ewes, before maternal recognition of pregnancy, but did not seem to disturb reproductive function.

No increase in pregnancy rate of mares after preovulatory deep uterine horn application of misoprostol

A potential source of fertility loss in mares is oviductal dysfunction, potentially caused by masses or debris in the lumen, that may prevent either sperm from reaching the fertilization site or the embryo from reaching the uterus. Recently a novel therapeutic method leading to increased pregnancy results was described by infusing misoprostol, a synthetic prostaglandin E1, in the uterus of mares with unexplained fertility problems. In this study, we aimed, after examining the compatibility of misoprostol with semen, to evaluate the pregnancy rate after routine preovulatory deep uterine horn application of misoprostol in clinically normal oestrous mares, which were inseminated in the same cycle. In experiment 1, ejaculates of 10 stallions diluted with INRA 96™ were mixed with different concentrations of misoprostol (0.01 mg/mL, 0.001 mg/mL, 0.0001 mg/mL, and 0.00001 mg/mL) and total semen motility was evaluated immediately, 12, 24, 48, and 72 h later, and compared with a control sample (mixed with NaCl 0.9%). In experiments 2 and 3, 33 privately-owned clinically normal oestrous mares were each allocated to a treatment or control group. Ovulation was then induced with intramuscularly 2.25 mg deslorelin acetate. At the moment of ovulation induction (experiment 2) and 24 h earlier (experiment 3), 0.2 mg misoprostol diluted in 2 mL NaCl 0.9% were applied deep in the uterine horn (treatment groups) and pure 2 mL NaCl 0.9% in the mares of the control groups. Mares were then inseminated 24 h after deslorelin administration and prior to ovulation with commercial chilled-warmed or frozen-thawed semen, as well as immediately after ovulation detection (both types of semen) maximally 48 h after ovulation induction. In experiment 1, regardless of time and compared with the control groups, all solutions with different concentrations of misoprostol had a negative effect on total motility of semen, which was significant for the highest concentrations (0.01 mg/mL: 18.0% reduction, CI = 22–13%, p = < 0.01). We found no beneficial effect of preovulatory uterine treatment with misoprostol on pregnancy rate (OR = 0.45, CI = 0.15–1.31, p = 0.14): in experiment 2, 2/11 (18.2%) mares of the treatment group became pregnant vs. 12/22 (54.5%) mares in the control group (OR = 0.19, CI = 0.03–1.06, p = 0.07), in experiment 3, 5/14 (35.7%) mares in the treatment group vs. 7/19 (36.8%) mares in the control group (OR = 0.95, CI = 0.23–4.02, p = 0.95), respectively. In conclusion, pregnancy rate was not increased in reproductively normal mares with routine preovulatory deep uterine horn application of misoprostol.

Doppler ultrasonographic evaluation of medically treated female dogs with cystic endometrial hyperplasia–pyometra complex

Monitoring the treatment outcome of canine cystic endometrial hyperplasia-pyometra complex (CEH-P) is essential to identify nonresponding patients who might require a rapid intervention to avoid life-threatening conditions. Uterine artery Doppler characterization may contribute to monitoring medically treated CEH-P, but published studies are currently lacking. Therefore, the aim of this longitudinal prospective observational study was to evaluate uterine artery resistance changes in the medical treatment of female dogs with CEH-P. Twelve diestrous female dogs suffering from CEH-P were treated with a combined protocol of cabergoline, cloprostenol, and antibiotics. The animals were clinically and ultrasonographically evaluated before the beginning of treatment (day 0) and on days 3, 7, 14, 21, and 28. The widest transverse diameter and luminal diameter of uterine horns were measured, as well as the peak systolic velocity (PSV) and end diastolic velocity (EDV) of uterine arteries. The resistance index (RI), pulsatility index (PI), and notch deep index (NDI) were calculated. On day 3, nine of 12 animals showed clinical improvement. In all these female dogs, the widest transverse sectional diameter (P<0.01), luminal diameter (P<0.01), PSV (P<0.01), and EDV (P<0.01) progressively decreased, while RI (P<0.01), PI (P<0.01), and NDI (P<0.01) increased up to day 21. In nonresponding dogs (3/12),Dopplerparameters remained unchanged. Although comparisons of PSV, EDV, and NDI were significantly different, these results should be cautiously interpreted due to the low statistical power. female dogs that responded to this treatment showed an increase in uterine artery resistance, along with clinical and ultrasonographic improvement.

Maternal, placental, and fetal distribution of titanium after repeated titanium dioxide nanoparticle inhalation through pregnancy

Epidemiological studies have associated ambient engineered nanomaterials or ultrafine particulate matter (PM0.1), collectively referred to as nanoparticles (NPs), with adverse pregnancy outcomes including miscarriage, preterm labor, and fetal growth restriction. Evidence from non-pregnant models demonstrate that NPs can cross the lung air-blood barrier and circulate systemically. Therefore, inhalation of NPs during pregnancy leading to fetoplacental exposure has garnered attention. The purpose of this study was to evaluate the distribution of inhaled titanium dioxide nanoparticles (nano-TiO2) from the maternal lung to maternal and fetal systemic tissues. Pregnant Sprague Dawley rats were administered whole-body exposure to filtered air or of nano-TiO2 aerosols (9.96 ± 0.06 mg/m3) between gestational day (GD) 4 and 19. On GD 20 maternal, placental, and fetal tissues were harvested then digested for ICP-MS analysis to measure concentrations of titanium (Ti). TEM was used to visualize particle internalization by the placental syncytium. The results demonstrate the extrapulmonary distribution of Ti to various maternal organs during pregnancy. Our study found Ti accumulation in the decidua/junctional and labyrinth zones of placentas embedded in all sections of uterine horns. Further, NPs deposited in the placenta, identified by TEM, were found intracellularly within nuclear, endoplasmic reticulum, and vesicle organelles. This study identified the systemic distribution and placental accumulation of Ti after nano-TiO2 aerosol inhalation in a pregnancy model. These findings arouse concerns for poor air quality for pregnant women and possible contributions to adverse pregnancy outcomes.

Pregnancy Patterns Impact Live Birth Rate for Patients With Intrauterine Adhesions After Hysteroscopic Adhesiolysis: A Retrospective Cohort Study.

ObjectiveThe pregnancy patterns and other factors of live birth for patients with intrauterine adhesions (IUAs) were identified by analyzing the clinical features of pre-, intra-, and post-hysteroscopic adhesiolysis (HA).DesignA total of 742 patients with IUAs who wanted to become pregnant underwent HA from January 2017 to May 2018 at the Third Xiangya Hospital of Central South University. The patient follow-up period was 2 years post-HA. A logistic regression was performed to analyze the clinical characteristics associated with a live birth for patients with IUAs. Pre-operative clinical indicators included age, gravidity, parity, abortion, IUA recurrence, menstrual patterns, disease course. Intraoperative clinical features assessed in the last operation were uterine cavity length, IUA appearance, IUA area, number of visible uterine cornua, number of visible tubal ostia, AFS scores. Pregnancy patterns were post-hysteroscopic adhesiolysis features.ResultsAmong the 742 IUA patients, 348 (46.9%) had a live birth and 394 (53.1%) did not. A bivariate and binary logistic regression analysis showed that IUA patients’ pregnancy patterns, age, number of visible tubal ostia noted by a second-look hysteroscopy, and American Fertility Society (AFS) scores were significantly related to the live birth rate (P < 0.05).ConclusionsPregnancy patterns, age, number of visible tubal ostia, and AFS scores were significantly related to the live birth rate and may be considered potential predictors of the live birth rate in IUA patients. The indications of assisted reproductive technology (ART) might be a better choice for patients with recurrent IUAs.