Hydroxysafflor Yellow A Inhibits Staphylococcus aureus-Induced Mouse Endometrial Inflammation via TLR2-Mediated NF-kB and MAPK Pathway.

The present study is designed to investigate the effect of hydroxysafflor yellow A (HYA) on Staphylococcus aureus (S. aureus)-induced mouse endometrial inflammation and to explore its molecular mechanism. We established a mouse endometritis model by intrauterine injection of S. aureus and intrauterine injection of HYA for treatment. Immunohistochemistry, immunofluorescence, and Western blot were used to detect protein expression in uterine tissue, and qPCR was used to measure mRNA expression. HYA could significantly weak uterine pathological changes caused by S. aureus and reduce MPO activity, CD45, CD3, and ED-1 protein expression in uterine tissues of S. aureus-infected mice. Similarly, HYA also significantly decreased S. aureus induced the increase in TNF-α, IL-1ß, and IL-6 in uterine tissue. In vivo, we found that knockdown of TLR2 was very important could significantly reduce S. aureus induced the elevated expression of TNF-α, IL-1ß, and IL-6 in mEECs. Importantly, in terine tissues of S. aureus-infected mice, HYA significantly decreased the ratio of p-p65/p65, p-IKBα/IKBα, p-p38/p38, p-Erk/Erk, and p-JNK/JNK expression. HYA displays anti-inflammatory effects on S. aureus mouse endometrial inflammation, and this effect might be related to HYA which could block TLR2-mediated NF-kB and MAPK pathway.

Endometritis in the bitch: Immunohistochemical localization of cyclooxygenase 2.

Background: In several mammals, subfertility or infertility associated with endometritis was reported. Although there have been studies about endometritis in bitches, the pathophysiological mechanisms are not completely known. Aim: This study aimed to evaluate the immunohistochemical expression of Cyclooxygenase 2 (COX2) in clinically healthy bitches with normal uterine tissue and bitches with endometritis. Methods: Forty-eight mixed breed bitches in diestrus were used. Uterine biopsies were collected for diagnosis [normal endometrium (n = 15; NE), cystic endometrial hyperplasia (n = 1), atrophy (n= 2), acute endometritis (n = 9; AE), subacute endometritis (n = 7; SE), and chronic endometritis (n = 14; CE)]. Immunostaining and quantification of positively stained cells was performed on full-thickness uterine biopsies. Data were analyzed by the GLIMMIX procedure of SAS. Results: COX2 immunostaining was scattered and restricted to cells in the stroma in bitches with NE. However, in bitches with endometritis, strong staining was observed in the luminal epithelium, glandular epithelium, and stromal cells. Staining was also observed in inflammatory cells localized in the stroma as well as inside of the glands. The percentage of COX2 positive stromal cells in bitches with AE, SE, and CE was significantly higher compared with NE (p < 0.005). In addition, the percentage of COX2 positive stromal cells in bitches with SE, and CE was significantly lower compared with AE (p < 0.003). Conclusion: COX2 could be involved in the pathophysiological mechanisms producing endometritis without the presence of cystic endometrial hyperplasia in bitches. However, further researches on this topic are required.

A functional role for AMPK in female fertility and endometrial regeneration.

Adenosine monophosphate-activated protein kinase (AMPK) is a highly conserved heterotrimeric complex that acts as an intracellular energy sensor. Based on recent observations of AMPK expression in all structures of the female reproductive system, we hypothesized that AMPK is functionally required for maintaining fertility in the female. This hypothesis was tested by conditionally ablating the two catalytic alpha subunits of AMPK, Prkaa1 and Prkaa2, using Pgr-cre mice. After confirming the presence of PRKAA1, PRKAA2 and the active phospho-PRKAA1/2 in the gravid uterus by immunohistochemistry, control (Prkaa1/2 fl/fl ) and double conditional knockout mice (Prkaa1/2 d/d ) were placed into a six-month breeding trial. While the first litter size was comparable between Prkaa1/2 fl/fl and Prkaa1/2 d/d female mice (P = 0.8619), the size of all subsequent litters was dramatically reduced in Prkaa1/2 d/d female mice (P = 0.0015). All Prkaa1/2 d/d female mice experienced premature reproductive senescence or dystocia by the fourth parity. This phenotype manifested despite no difference in estrous cycle length, ovarian histology in young and old nulliparous or multiparous animals, mid-gestation serum progesterone levels or uterine expression of Esr1 or Pgr between Prkaa1/2 fl/fl and Prkaa1/2 d/d female mice suggesting that the hypothalamic-pituitary-ovary axis remained unaffected by PRKAA1/2 deficiency. However, an evaluation of uterine histology from multiparous animals identified extensive endometrial fibrosis and disorganized stromal-glandular architecture indicative of endometritis, a condition that causes subfertility or infertility in most mammals. Interestingly, Prkaa1/2 d/d female mice failed to undergo artificial decidualization. Collectively, these findings suggest that AMPK plays an essential role in endometrial regeneration following parturition and tissue remodeling that accompanies decidualization.

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding-induced endometritis and the effect of a specific iNOS inhibitor in vitro.

Emerging research suggests that the nitric oxide system may play a role in persistent breeding-induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose-dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post-challenge and cultured at 37°C for 24 hr in L-arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non-parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.

Associations of periparturient plasma biochemical parameters, endometrial leukocyte esterase and myeloperoxidase, and bacterial detection with clinical and subclinical endometritis in postpartum dairy cows.

This study was aimed at demonstrating associations between peripheral biochemical parameters, endometrial leukocyte esterase (LE) and myeloperoxidase (MPO), and bacterial detection with the degree of endometrial inflammation, and determining the best time postpartum for diagnosing endometritis to predict subsequent fertility in dairy cows. Plasma albumin, blood urea nitrogen (BUN), total cholesterol (T-cho), NEFA, and BHBA concentrations were analyzed in 43 Holstein cows at 3, 5 and 7 weeks postpartum (W3, W5 and W7). Endometrial samples were collected at W3, W5 and W7 to examine LE and MPO activities, bacterial detection rates, and PMN% profiles. The 43 cows were divided into healthy (HE), subclinical endometritis (SE), and clinical endometritis (CE) groups, classified differently at W3, W5 and W7 based on the definitions of SE and CE for each of the three weeks pp. LE level had an association with PMN% in all weeks pp (P<0.05). Albumin and BUN levels had weak negative associations with endometrial PMN% at W3. Pathogenic bacterial detection rates were higher in the cows with endometritis at W3 and W5. Conception rate at first artificial insemination tended to be lower (P=0.057) in the cows diagnosed with endometritis at W3 than in the healthy cows. In conclusion, associations were found between endometrial LE and endometritis, but not for MPO and endometritis. Diagnosing endometritis in W3 may be the best moment to predict subsequent fertility.

Prostaglandin synthesis enzymes' gene transcription in bitches with endometritis.

Endometritis is a major cause of infertility in many domestic species. However, until now the pathogenesis of the endometritis in the bitch is unclear. The aim of this study was to evaluate the gene transcription pattern of prostaglandin (PG) synthesis enzymes (cyclooxygenase [COX2], PTGES-1 and PGFS) in the endometrium of bitches with or without endometritis. Thirty mixed breed bitches in dioestrus, aged between 1 and 5 years, and weighing between 10 and 30 kg were used. After ovariohysterectomy (OVX), uterine biopsy samples were collected from the middle part of both horns. Then, endometrial epithelium was collected using the cytobrush method and mRNA analysis was performed by real-time RT-PCR. Data were analysed with Kruskal-Wallis anova using the sas® software. Uterine condition was identified by endometrial biopsies (normal endometria [n = 11; NE], acute endometritis [n = 10; AE] and chronic endometritis [n = 9; CE]). The COX2, PTGES-1 and PGFS/AKR1C3 mRNA expression in bitches with and without endometritis was similar. Except for PGFS/AKR1C3, gene transcription of COX2 and PTGES-1 was significantly increased in AE compared with CE. In addition, COX2 gene transcription was significantly increased in AE compared with NE. In contrast, no differences were found for COX2, PTGES-1 and PGFS/AKR1C3 mRNA expression in the samples of NE compared with CE.

Expression of inflammation-associated genes in circulating leukocytes and activity of indoleamine-2,3-dioxygenase in dairy cattle with acute puerperal metritis and bacteremia.

OBJECTIVE: To investigate whether expression of genes associated with inflammation and activity of indoleamine-2,3-dioxygenase (IDO) correlated with disease status and prevalence of bacteremia in post-partum dairy cattle with and without acute puerperal metritis (APM). PROCEDURES: Blood was collected from cattle with APM and control cattle matched by parity and days in milk. Leukocytes were isolated and expression of 6 genes was quantified. Activity of IDO was measured in serum with higher performance liquid chromatography (HPLC). RESULTS: The relative expression of IL-1ß in cattle with APM was significantly lower than that in controls. IDO activity was not significantly different between bacteremic and non-bacteremic cattle CONCLUSIONS AND CLINICAL RELEVANCE: The expression of IL-1ß was lower in cattle with APM. The lower levels of IL-1ß expression in PBMCs of cattle with APM suggest impaired inflammatory responses and may contribute to the development of the disease in this population of animals.

Successful implantation after reducing matrix metalloproteinase activity in the uterine cavity.

BACKGROUND: Recently, the concept of recurrent implantation failure (RIF) in assisted reproductive technology has been enlarged. Chronic uterine inflammation is a known cause of implantation failure and is associated with high matrix metalloproteinase (MMP) activity in uterine cavity flushing. MMP activity of women with RIF has been reported to be higher than that of fertile women. In the present retrospective study we evaluated the efficacy of treatment for high MMP activity in the uterine cavity of patients with RIF. METHODS: Of the 597 patients recruited to the study, 360 patients underwent MMP measurements and 237 patients did not (control group). All patients had failed to become pregnant, despite at least two transfers of good-quality embryos. Gelatinase MMP-2 and MMP-9 activity in uterine flushing fluid was detected by enzymology (MMP test). All samples were classified into two groups (positive or negative) based on the intensity of the bands on the enzyme zymogram, which represents the degree of MMP activity. Patients who tested positive on the initial test were treated for 2 weeks with a quinolone antibiotic and a corticosteroid, and subsequently underwent a second MMP test. Negative results on the second MMP tests after treatment and subsequent rates of pregnancy and miscarriage were used to evaluate the efficacy of treatment. Data were analyzed by the Mann-Whitney U-test and the chi-square test. RESULTS: Of the patients who underwent the MMP test, 15.6% had positive results (high MMP activity). After treatment, 89.3% of patients had negative results on the second MMP test. These patients had a significantly better pregnancy rate (42.0%) than the control group (26.6%), as well as a lower miscarriage rate (28.5% vs 36.5%, respectively). CONCLUSIONS: A 2-week course of antibiotics and corticosteroids effectively improves the uterine environment underlying RIF by reducing MMP activity.

Creatine kinase as an indicator for hysterectomy in postpartum endomyometritis due to group A streptococci: a hypothesis illustrated by a case report.

BACKGROUND: Puerperal group A streptococcus (GAS) infection, once the leading cause of postpartum sepsis, has been increasing again since the 1980s. Streptococcal toxic shock syndrome (STSS) is a serious complication characterized by rapidly spreading GAS infection, shock, and multiple organ failure. Immediate recognition and implementation of therapy is crucial for survival. Making informed decisions regarding surgical debridement, namely hysterectomy, based on clinical indicators is difficult for practitioners. OBJECTIVES: This article discusses the potential role of creatine kinase in the decision-making process for treatment of STSS, particularly with regard to hysterectomy. MATERIAL AND METHODS: A case report is presented. The literature was searched using the key words 'group A streptococcus', 'postpartum hysterectomy', 'creatine kinase', 'endomyometritis', and 'streptococcal toxic shock syndrome' in PubMed and the UptoDate database. Relevant articles published between 1991 and 2011 were evaluated. CONCLUSION: Decisions regarding hysterectomy in STSS management are difficult. A rise in CK levels in the serum may indicate involvement of the myometrium and may be an important parameter in the difficult decision of hysterectomy when treating STSS.

Expression of cyclooxygenase-2 in the endometrium of gilts with different stages of endometritis.

The present study determined the association among the expression of COX-2, stages of endometritis, and types and number of local immune cells infiltrating into the gilts' endometrium. The uterine tissues from 24 Landrace x Yorkshire gilts identified as acute endometritis (n = 7), chronic endometritis (n = 7), and normal endometrium (n = 10) were included. The tissues were prepared for both histological and immunohistochemical investigations. The immunoexpression of COX-2 in every layer of the gilts' endometria was appraised by avidin-biotin-peroxidase complex method via image analysis; and was reported as percentage of positive area and staining index. The results revealed that the immunoexpression of COX-2 was found only in the surface epithelial layer. The gilts with acute endometritis possessed higher both percentage of positive area (68.99% versus 4.50% and 3.43%, P < 0.001) and staining index (1.13 versus 0.05 and 0.04, P < 0.001) than those with chronic endometritis and normal endometrium, respectively. Positive correlations between the number of surface epithelial neutrophils and percentage of COX-2 positive area (r = 0.47, P = 0.022), as well as mean staining index (r = 0.44, P = 0.032) were observed. In conclusion, the immunoexpression of COX-2 was found strongest in the gilts with acute endometritis, meanwhile it was not different between those with chronic endometritis and normal endometrium. This suggested that the expression of COX-2 might be dependent not only on the infiltration of local immune cells in the endometrium, but also on the duration of exposure with inflammatory agents.

Significance of nitric oxide concentration in plasma and uterine secretes with puerperal endometritis in dairy cows.

Endometritis is an inflammation of the endometrial lining of the uterus without systemic signs, which is associated with chronic postpartum infection of the uterus with pathogenic bacteria. Nitric oxide (NO) is an inflammatory mediator that among other effects causes smooth muscle relaxation and mediated cytoimmunity and inflammation toxicity. To see if the nitric oxide concentration in plasma and uterine secrets is related with postpartum endometritis, NO concentrations in plasma and uterine secrets were measured in dairy cows with puerperal endometritis (clinical endometritis (n = 60) and subclinical endometritis (n = 58)). Cows with clinical or subclinical endometritis showed higher concentrations of NO in both plasma and uterine secrets when compared with normal cows and the highest concentrations of NO in plasma and uterine secrets were found in dairy cows with clinical endometritis. Expression level of NOS2 mRNA in endometrial biopsies from cows with puerperal endometritis was also higher and the highest expression of NOS2 mRNA was found in cows with clinical endometritis. The results showed that concentrations of NO in plasma and uterine fluid are related with the degree of endometritis which may be useful to diagnose the endometritis in dairy cows.

Blood COX-2 and PGES gene transcription during the peripartum period of dairy cows with normal puerperium or with uterine infection.

In the dairy cow, puerperal uterine intra-luminal concentrations of PGE(2) are related to the establishment and severity of uterine infections. Here we evaluated whether the blood concentrations of PGE(2) and the gene transcription profiles of enzymes involved in its synthesis (cyclooxygenase-2 and prostaglandin E synthase) could be used as markers of predisposition and/or presence of puerperal uterine infections. We also studied the relationship between the endocrine status and the leukocyte profiles around parturition and the transcription patterns of the genes. Finally, we have characterized the in vitro gene transcription and expression response to a challenge of LPS. Gene transcription profiles, quantified by real-time PCR, were similar in normal puerperium and metritis/endometritis cows, indicating that they are not suitable markers of predisposition to/presence of puerperal uterine infections. Transcription decreased from 2 weeks before parturition until parturition, when a minimum was attained, and then increased during the first week postpartum. The lowest gene transcription, at parturition, was coincidental with the highest total leukocytes, polymorphonuclear neutrophils and CD14 positive cell numbers. It is suggested that by this mechanism, a large number of PMN can be recruited into the uterus after parturition, avoiding an excessive acute inflammatory response. The lowest gene transcription was also coincidental with the surge in cortisol concentrations, indicating that this hormone plays a main immunomodulatory role around parturition. Gene transcription was significantly greater after stimulation with LPS than in non-stimulated blood. We suggest that this PGE(2) producing cells might arrive to the uterine lumen, contributing to the local PGE(2) concentrations and mediating the inflammatory response.

Experimental endometriosis in the rat is correlated with colonic motor function alterations but not with bacterial load.

Endometriosis commonly presents with symptoms that mimic chronic gastrointestinal disorders. The authors used the autotransplantion model of endometriosis in rats to investigate the possible underlying mechanisms. After the rats were killed, the presence of endometriotic vesicles, colonic inflammation, and white blood cell (WBC) numbers in the peritoneal fluid was determined. Sections of colon and of jejunum were collected for measurement of myeloperoxidase (MPO) activity and bacterial counts, and isometric recording in response to acetylcholine was measured in segments of longitudinal and circular smooth muscle. Experimental animals had significantly more colonic damage, MPO activity, and WBC numbers than controls did. There was no significant difference in the total bacterial load; however, experimental animals demonstrated an increased tension in the longitudinal muscle, which correlated with WBC numbers and colonic damage. In summary, this study presents evidence for a significant effect of peritoneal endometriosis on colonic function and integrity, which may help explain the gastrointestinal symptoms associated with this disease.

Serum activity of matrix metalloproteinases -2 and -9.

Matrix metalloproteinases belong to the key effectors of tissue remodeling in health and disease. Matrix metalloproteinases -2 and -9, the most prevalent representatives of this family, are expressed in the endometrium. Chronic endometritis concomitant with sterility and spontaneous abortions is associated with decreased content of matrix metalloproteinases in the endometrium. Chronic endometritis combined with sterility correlates with decreased serum activities of matrix metalloproteinases -2 and -9, which returns to normal after recovery. Measurements of serum activities of matrix metalloproteinases -2 and -9 are proposed for monitoring of the dynamics and treatment efficiency in chronic endometritis.

Matrix metalloproteinases (MMPs) in the endometrium of bitches.

The relationships between activities of matrix metalloproteinases (MMPs) in the canine uterus and the occurrence of degeneration of the luminal epithelium, cystic endometrial hyperplasia, pyometra and uterine remodelling post partum were determined. Mature bitches (n = 10) were ovariectomized, treated with hormones (oestradiol benzoate, progestagen) and investigated at stages simulating pro-oestrus (n = 2), oestrus (n = 2), dioestrus (n = 2), and mid- (n = 2) and late (n = 2) anoestrus (3 and 9 weeks, respectively, after cessation of treatment with progestagen). Untreated bitches (n = 1 per group) served as controls (Expt 1). An additional 10 ovariectomized bitches, at the end of treatment-induced simulated dioestrus, were treated each day for a further 3 weeks either with the same dose (standard dose, n = 3), a decreased dose (n = 3) or an increased dose (n = 3) of progestagen, or no treatment (withdrawal dose, n = 1). These bitches were then investigated (Expt 2). A suture was placed in the lumen of one uterine horn of another five bitches at ovariectomy. Three of these bitches were treated to induce simulated dioestrus and two bitches served as untreated controls. In the hormone-treated bitches, the suture resulted in cystic endometrial hyperplasia in one bitch and in cystic endometrial hyperplasia with pyometra in two bitches. The control bitches showed no cystic endometrial hyperplasia or pyometra (Expt 3). Four intact bitches were studied at 2 (n = 1), 3 (n = 2) and 11 (n = 1) weeks post partum. Uterine tissues were also collected from two bitches with naturally occurring cystic endometrial hyperplasia with pyometra (Expt 4). All uteri were examined histologically and the activities of MMP-2, -7 and -9 (latent and active forms) were assessed using zymography of extracts of endometrium. In Expts 1 and 2, marked degeneration of the luminal epithelium in six of 25 bitches (simulated mid-anoestrus, withdrawal dose and decreased dose groups) was not associated with changes in MMP activities. Markedly increased activities of MMP-2 (active form), -7 (latent form) and -9 (active and latent forms) were observed in the bitches with cystic endometrial hyperplasia with pyometra (but not with cystic endometrial hyperplasia alone) and in the bitches at 2 and 3 weeks post partum (but not at 11 weeks post partum). These results indicate that MMPs are not involved with degeneration of the luminal epithelium, but are involved with uterine remodelling in the postpartum bitch and with cystic endometrial hyperplasia with pyometra.

The characteristic pattern of aspartate aminotransferase and alanine aminotransferase in the bitch with the cystic hyperplasia-pyometra complex: effect of medical or surgical treatment.

In 75 clinically normal unspayed female control dogs between two and eleven years old the average plasma level of aspartate aminotransferase (AST) was 21.6 +/- 5.7 (+/- SD) IU/l, of alanine aminotransferase (ALT) 40.4 +/- 13.0 IU/l and of the AST/ALT ratio 0.6 +/- 0.2. These values showed only minor changes over years. In 96 bitches with the cystic hyperplasia-pyometra complex there was a very significant increase of the AST, decrease of the ALT and increase of the AST/ALT ratio. The changes were more pronounced in 62 clinically ill bitches with typical endometritis post oestrum, in 18 dogs with gram negative organisms in the uterus and in 53 bitches with white blood cell (WBC) levels higher than 40 X 10(9)/1. Renal failure had no influence on the specific changed values. The changed values returned either temporarily to normal after prostaglandin (PGF2 alpha)-treatment or definitely after ovariohysterectomy.

Distribution of peroxidase and granulocytes in the human uterus.

A variety of uterine cell types demonstrate endogenous peroxidase activity. Ultracytochemical localization, biochemical assays, and uterine granulocyte counts were used to characterize peroxidase activity in various regions of the human uterus and cervix during the menstrual cycle and during the postmenopausal period. Previous studies of rat uteri, using electron microscopy and biochemical assays, have shown that endometrial peroxidase is induced by estrogenic stimulation (Anderson, De Sombre, and Kang, J Cell Biol 64:668, 1975; and Biol Reprod 16:409, 1977). Tissue samples from four regions of the human uterus and one sample from the endocervix were processed for ultrastructural cytochemistry, biochemical assay, and histology. Endogenous peroxidase activity was identified with electron microscopy in the endoplasmic reticulum of endometrial epithelial cells lining four regions of the uterine cavity; the isthmus, body (2), and fundus, of some proliferative phase (2 of 6), all secretory phase (4 of 4) and all postmenopausal (3 of 3) endometria. Peroxidase activity was not demonstrable in endocervical epithelial cells. Endogenous peroxidase activity was also identified in the cytoplasmic granules of uterine eosinophils and neutrophils and in the endoplasmic reticulum of mast cells. These uterine granule-containing cells, identified with special stains in the histologic sections, were quantitated. Approximately 80% of these "uterine granulocytes" from normal uteri without intrauterine devices were neutrophils. In women of reproductive age the uterine granulocytes, although present throughout the menstrual cycle, were most numerous in the endocervix and lower uterine segment. The highest biochemical assays of peroxidase activity were also obtained in the cervix and lower uterine segment. Uterine granulocyte counts varied directly with biochemical assays of peroxidase activity indicating that they were a major determinant of biochemical peroxidase activity. Endometrial epithelial peroxidase is anatomically and temporally well placed to function as an important adjunct in maintaining a mucosal barrier to microorganisms.

Histochemical demonstration of lysosomal hydrolase activity in endometrial mononuclear cells. II. Abnormal endometrium.

The mononuclear cells in the endometrial stoma change in reactivity for lysosomal hydrolases during the menstrual cycle. Lymphoid follicles may occur in the stroma in any phase of the cycle and have been found in gestational endometrium. However, these cells have no significant lysosomal activity. Alterations in the endometrium are reflected in modified patterns of activity. Endometritis, association with an intrauterine contraceptive device, pregnancy, and adenocarcinoma result in increased numbers and staining intensity of mononuclear cells. In contrast, no consistent changes were apparent in foci of glandular hyperplasia, and decreased staining was seen in atrophic areas of endometrium. These data suggest that interstitial mononuclear cells are a sensitive monitor of morphologic changes in the endometrium.