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Background The use of diuretics in patients on haemodialysis (HD) is thought to maintain diuresis. However, this assumption and the optimal dose are based on little scientific evidence, and associations with clinical outcomes are unclear. Methods We reported international variations in diuretic use and loop diuretic dose across 27 759 HD patients with dialysis vintage <1 year in the Dialysis Outcomes and Practice Patterns Study phases 2–5 (2002–2015), a prospective cohort study. Doses of torsemide (4:1) and bumetanide (80:1) were converted to oral furosemide-equivalent doses. Adjusted Cox, logistic and linear regressions were used to investigate the association of diuretic use and dose with outcomes. Results Diuretic utilization varied widely by country at vintage <3 months, ranging from >80% in Germany and Sweden to <35% in the USA, at a median dose ranging from 400–500 mg/day in Germany and Sweden to <100 mg/day in Japan and the USA. Neither diuretic use nor higher doses were associated with a lower risk of all-cause mortality, a higher risk of hospitalization for fracture or elevated parathyroid hormone levels, but the prescription of higher doses (>200 mg/day) was associated with a higher risk of all-cause hospitalization. Conclusions Substantial international differences exist in diuretic prescriptions, with use and doses much higher in some European countries than the USA. The prescription and higher doses of loop diuretics was not associated with improved outcomes. [ABSTRACT FROM AUTHOR]

Background Chronic kidney disease is associated with a high cardiovascular risk. Compared with glomerular filtration rate–matched CKD patients (CKDps), we previously reported a 2.7-fold greater risk of global mortality among kidney transplant recipients (KTRs). We then examined aortic stiffness [evaluated by carotid–femoral pulse wave velocity (CF-PWV)] and cardiovascular risk in KTRs compared with CKDps with comparable measured glomerular filtration rate (mGFR). Methods We analysed CF-PWV in two cohorts: TransplanTest (KTRs) and NephroTest (CKDps). Propensity scores were calculated including six variables: mGFR, age, sex, mean blood pressure (MBP), body mass index (BMI) and heart rate. After propensity score matching, we included 137 KTRs and 226 CKDps. Descriptive data were completed by logistic regression for CF-PWV values higher than the median (>10.6 m/s). Results At 12 months post-transplant, KTRs had significantly lower CF-PWV than CKDps (10.1 versus 11.0 m/s, P = 0.008) despite no difference at 3 months post-transplant (10.5 versus 11.0 m/s, P = 0.242). A lower occurrence of high arterial stiffness was noted among KTRs compared with CKDps (38.0% versus 57.1%, P < 0.001). It was especially associated with lower mGFR, older age, higher BMI, higher MBP, diabetes and higher serum parathyroid hormone levels. After adjustment, the odds ratio for the risk of high arterial stiffness in KTRs was 0.40 (95% confidence interval 0.23–0.68, P < 0.001). Conclusions Aortic stiffness was significantly less marked in KTRs 1 year post-transplant than in CKDps matched for GFR and other variables. This observation is compatible with the view that the pathogenesis of post-transplant cardiovascular disease differs, at least in part, from that of CKD per se. [ABSTRACT FROM AUTHOR]

Background Optimal parathyroid hormone (PTH) control during non-dialysis chronic kidney disease (ND-CKD) might decrease the subsequent risk of parathyroid hyperplasia and uncontrolled secondary hyperparathyroidism (SHPT) on dialysis. However, the evidence for recommending PTH targets and therapeutic strategies is weak for ND-CKD. We evaluated the patient characteristics, treatment patterns and PTH control over the first year of haemodialysis (HD) by PTH prior to HD initiation. Methods We studied 5683 incident HD patients from 21 countries in Dialysis Outcomes and Practice Patterns Study Phases 4–6 (2009–18). We stratified by PTH measured immediately prior to HD initiation and reported the monthly prescription prevalence of active vitamin D and calcimimetics over the first year of HD and risk of PTH >600 pg/mL after 9–12 months on HD. Results The 16% of patients with PTH >600 pg/mL prior to HD initiation were more likely to be prescribed active vitamin D and calcimimetics during the first year of HD. The prevalence of PTH >600 pg/mL 9–12 months after start of HD was greater for patients who initiated HD with PTH >600 (29%) versus 150–300 (7%) pg/mL (adjusted risk difference: 19%; 95% confidence interval : 15%, 23%). The patients with sustained PTH >600 pg/mL after 9–12 months on HD were younger, more likely to be black, and had higher serum phosphorus and estimated glomerular filtration rates at HD initiation. Conclusions Increased PTH before HD start predicted a higher PTH level 9–12 months later, despite greater use of active vitamin D and calcimimetics. More targeted PTH control during ND-CKD may influence outcomes during HD, raising the need for PTH target guidelines in these patients. [ABSTRACT FROM AUTHOR]

Background MYH9-related diseases (MYH9-RD) are autosomal dominant disorders caused by mutations of the MYH9 gene encoding the non-muscle myosin heavy chain IIA. They are characterized by congenital thrombocytopenia, giant platelets and leucocyte inclusions. Hearing impairment, pre-senile cataract and nephropathy can also occur. We aimed to evaluate renal involvement and outcome in MYH9-RD patients followed-up by nephrologists. Methods We conducted a retrospective multicentre observational study of 13 patients among 9 families with MYH9 mutation diagnosed by genetic testing and immunofluorescence assay referred to nephrologists. Results At initial referral, median age was 30 (range 14–76) years. Median estimated glomerular filtration rate was 66 mL/min/1.73 m2 (0–141) and two patients had already end-stage renal disease (ESRD). Renal presentation associated proteinuria (n  = 12), haematuria (n  = 6) and hypertension (n  = 6). Three patients developed a rapid onset ESRD whereas five others had a relatively stable kidney function over a 3-year median follow-up (1–34). Extra-renal features varied widely, with hearing impairment in six patients, cataract in two and mild liver dysfunction in seven. Thrombocytopenia existed at referral in 11 patients. Time to diagnosis varied from 0 to 29 years (median 3 years). Initial diagnoses such as idiopathic thrombocytopenic purpura (n  = 4) and focal segmental glomerulosclerosis (n  = 1) led to corticosteroid administration (n  = 4), intravenous immunoglobulins (n  = 3), cyclophosphamide (n  = 1) and splenectomy (n  = 1). Conclusions Renal involvement and outcome in MYH9-RD are heterogeneous. The diagnosis is often delayed and misdiagnoses can lead to unnecessary treatments. MYH9-RD should be considered in any patient with glomerular involvement associated with a low or slightly decreased platelet count and/or hearing loss and liver dysfunction. [ABSTRACT FROM AUTHOR]

Of note, urine osmolality and creatinine levels were not significantly higher in these patients, suggesting that the presence of urinary crystals was not primarily due to higher urine concentration. Patients with uric acid crystals had more frequently ongoing AKI and ongoing bacterial infection, and displayed higher plasma uric acid levels, higher urine uric acid levels, lower urinary pH and lower urinary citrate levels (Table 2). Previous literature has raised the interest of urine microscopy examination in order to detect or predict AKI and/or sepsis in critically ill patients [2], but none has described urinary crystals. [Extracted from the article]

Stem cells are functionally defined by their ability to self-renew and generate a progeny capable of creation or reconstitution of various tissues. Microarray analysis has shown a member of the transforming growth factor (TGF)-β superfamily, Lefty, to be the single most abundant inhibitor in stem cells and in maternal decidua that supports embryo implantation. Lefty is regulated by pathways such as Smad (Sma and Mad [mothers against decapentaplegic]) and WNT (wingless-type) and by the transcriptional factor Oct3/4 (octamer-binding transcription factor 3/4), which support ‘stemness.’ Lefty is also induced upon exit from the state of stemness, including forced in vitro differentiation, and leukemia inhibitory factor withdrawal. Lefty is a candidate in cell-fate decisions because of its unique ability to modulate the expression of TGF-β family proteins such as Nodal and by blanket inhibition of the activity of members of this family which require EGF-CFC (epidermal growth factor-Cripto, Frl-1, and Cryptic) as a coreceptor. [ABSTRACT FROM AUTHOR]

BACKGROUND: Endometrium is a unique tissue that is prepared for implantation of blastocyst during each menstrual cycle. In humans, if implantation does not occur or fails, endometrium is shed. METHODS and RESULTS: We identified ebaf/lefty, as a key cytokine, highly expressed in human endometrium during the non-receptive phase of tissue remodelling. Lefty was increased in the endometria of a number of patients with ‘unexplained infertility’ during the receptive phase, suggesting dysregulation of lefty as a potential factor contributing to infertility. Here, we showed that induction of a similar state of lefty overexpression in endometrium, by in vivo gene delivery, decreased implantation in pregnant mice. This state of overexpression could be induced by a retroviral vector transducing lefty or by liposome-mediated introduction of a lefty expression vector. Analysis of endometrium showed increased lefty after in vivo gene transfer. CONCLUSION: These findings suggest that induction of a state of lefty overexpression in endometrium leads to reduced implantation. [ABSTRACT FROM AUTHOR]

BACKGROUND: Reproductive tissues are unique structures that exhibit cyclic stromal remodelling during menstrual cycles in humans. Ebaf/lefty participates in tissue remodelling of human endometrium by induction of matrix metalloproteases (MMP).METHODS: We describe the temporal expression and spatial distribution of lefty and tissue remodelling events in mouse endometrium. RTPCR and real-time PCR were used to identify mRNA expression and western blots to analyse Lefty protein. Immunolocalization was performed with specific antibodies and horseradish peroxidase staining. RESULTS: Lefty was expressed in endometrium throughout the estrous cycle. Expression of MMP (MMP-2, -3, -7 and -14) was higher at estrus, metestrus and/or diestrus while collagen content of endometrium decreased in these phases. During pregnancy, lefty levels were higher on days 35 and were minimal by day 9. Similarly, expression of endometrial MMP was higher on days 3 and 5 of pregnancy and was low on day 9. During pregnancy, loss of collagen was initiated on day 3, persisted to day 5, and led to a significantly reduced collagen on day 9. Immunoreactive lefty decorated basal laminae, and was associated with extracellular matrix in stroma. CONCLUSIONS: Regulated expression and spatial distribution of lefty in mouse endometrium confines its biological impact on tissues that undergo remodelling during estrous cycle and pregnancy. [ABSTRACT FROM AUTHOR]

A well-orchestrated sequence of events enables the human endometrium to become receptive to embryo implantation during a defined period in the menstrual cycle. Ovarian steroids, oestradiol and progesterone, regulate many of these functions in a highly co-ordinated manner. There are no experimental systems for critically examining the regulation of endometrial functions by steroid hormones, especially those responses restricted to the epithelium. Using an experimental xenograft model where the steroid responses of normal endometrium could be predictably recapitulated, the role of progesterone in the regulation of α crystallin B in the glandular epithelial cells of human endometrium was established. [ABSTRACT FROM PUBLISHER]

During each menstrual cycle, the human endometrium undergoes a series of orchestrated and well controlled changes in anticipation of the arrival of the blastocyst. In the absence of implantation, the endometrium is shed. The underlying basis of the menstrual bleeding is not clear, however, it seems to be related to steroid hormone withdrawal. We showed that tumour necrosis factor-α (TNF-α) is released by human endometrium and that endometrial epithelial cells are a major source of TNF-α mRNA and protein. We show here that TNF-α mRNA shows a specific menstrual cycle-dependent expression. The expression of TNF-α is mostly minimal throughout the proliferative, early and mid-secretory phases. Expression of TNF-α mRNA, however, is increased in the human endometrium in the late secretory phase and during endometrial bleeding. Such a menstrual cycle-dependent expression suggests that specific signals regulate the expression of TNF-α mRNA in the human endometrium. In vitro, the expression of TNF-α mRNA in endometrial epithelial cells could be regulated by exogenous TNF-α. This induced expression was both time- and dose-dependent. In vitro, the TNF-α mRNA expression was not altered by oestrogen, progesterone, or both, in the endometrial epithelial cells under conditions that maintain the steroid hormone receptors. However, in vivo, oestrogen withdrawal led to an enhanced expression of TNF-α in endometrial epithelial cells. These findings suggest that the up-regulation of TNF-α in human endometrium in the late secretory phase may be related to the falling serum oestrogen concentration at the end of the menstrual cycle as well as the potentiating effect of released TNF-α on its own mRNA expression. [ABSTRACT FROM AUTHOR]

Human endometrium is the end organ of the hypothalamic-pituitary-ovarian axis. Therefore, endometrium is susceptible to changes in the cases of infertility that originate from disturbances in the normal functioning of this axis. In addition, some cases of unexplained infertility may be due to altered endometrial function. This disturbed endometrial function may originate from lesions in the molecular repertoire that are crucial to implantation. Human endometrium becomes receptive to implantation by the blastocyst within a defined period during the menstrual cycle. The duration of this so-called 'endometrial receptivity' or 'implantation' period seems to span from few days after ovulation to several days prior to menstruation. Successful implantation results from a co-ordinated series of events that would allow establishment of a timely dialogue between a receptive endometrium and an intrusive blastocyst. The members of the molecular repertoire that make endometrium receptive to implantation are gradually being recognized. Among these are the cytokines, integrins, heat shock proteins, tastin and trophinin. In addition, the expression of a second set of genes including tumour necrosis factor α (TNF-α) and ebaf, may be the appropriate signal for the closure of the 'implantation window', for making the endometrium refractory to implantation and for preparing it for the menstrual shedding. [ABSTRACT FROM PUBLISHER]

Apoptosis is a selective process for deletion of cells in various biological systems. This event, similar to proliferation, is tightly regulated, with both processes playing essential roles in the homeostasis of renewable tissues. In human endometrium, proliferation and apoptosis occur at opposing poles of the menstrual cycle. The proliferative phase is marked by rapid growth of the endometrial epithelial lining, whereas progressive increase in apoptosis in this tissue is the hallmark of the secretory and menstrual phases. The purpose of this review is to highlight some of the signals and molecular events which are associated with and that may participate in apoptosis. This is followed by a review of the current literature regarding apoptosis in human endometrium. [ABSTRACT FROM AUTHOR]

Human endometrium is a steroid-sensitive tissue and there is evidence that supports the viewpoint that heat shock proteins (HSP) are implicated in the regulation of steroid function. Therefore, in this study we examined the expression of various members of the heat shock family of proteins in the steroid-responsive human endometrium. Western blot analysis revealed that the expression of HSP90 showed minimal changes throughout the menstrual cycle. When normalized to the amount of HSP90, the expression of HSP27, HSP60 and the constitutive form of heat shock protein 70 (HSC70) increased progressively during the late proliferative and early secretory phases, and diminished in the mid- to late secretory and menstrual phases. In contrast, the inducible form of heat shock protein 70 (HSP70) did not undergo these changes. The cellular and subcellular localizations of these proteins were examined in human endometria by immunohistochemical staining. With the exception of HSP70, which was found primarily in the epithelial cells, the immunoreactivity for other heat shock proteins was found in both the stroraa and the epithelium. Immunoreactivity for HSP27 was found in the lymphoid aggregates within endometrial stroma, and both HSP27 and HSP90 were found in endothelial cells. The immunoreactive heat shock proteins were found in the nuclei and/or cytoplasm of cells. However, no consistent nuclear versus cytoplasmic staining emerged, and such localization was irrespective of the site, the cell type or the phase of the menstrual cycle. Our findings show that endometrium has a full complement of heat shock proteins. The menstrual cycle-dependent changes in the amounts of heat shock protein suggest regulation by steroid hormones. [ABSTRACT FROM PUBLISHER]

Granulocyte macrophage colony-stimulating factor (GM-CSF) has emerged as an important growth factor for trophoblast and other placental cells, leading to improved placental functioning and fetal survival. Recent observations have indicated that GM-CSF is synthesized by epithelial cells in the murine pregnant and non-pregnant uterus. In this study, the production of GM-CSF by cells derived from human endometrium is assessed using a sensitive bioassay and specific neutralization of the cytokine bioactivity with a monoclonal antibody to GM-CSF. Originally, GM-CSF was assayed in the culture supernatants of explant cultures of human endometria. Concentrations of GM-CSF up to 4440 pg/ml were detected. Subsequently, enriched epithelial cell cultures were prepared from glands isolated from human endometrium. The purity of epithelial cultures was demonstrated by the expression of cytokeratin, a weak immunoreactivity for vimentin and a lack of immunoreactivity for leukocyte common antigen, CD68, a macrophage-specific protein and endothelial marker (factor VIII-related antigens). Detected concentrations of GM-CSF were as high as 18,800 pg/ml. Furthermore, pure epithelial cells of a neoplastic endometrial cell line ECC1 secreted GM-CSF, confirming the ability of endometrial epithelial cells to secrete this cytokine. The immunostaining of dated endometria from proliferative and secretory phases showed primarily that epithelial cells, and to a lesser extent stromal cells, exhibited immunoreactivity for GM-CSF. A Western blot analysis, performed to validate the immunohistochemical data, confirmed the presence of an immunoreactive gene product for GM-CSF in human endometrium throughout the menstrual cycle. These findings indicate that human endometrium synthesizes GM-CSF and that epithelial cells are a major contributor to its production. [ABSTRACT FROM AUTHOR]

Implantation is a complex process requiring the interaction of the blastocyst, and subsequently the developing embryo with the endometrium. Initially, the detailed cellular interactions implicated in this process were defined. More recently, many signals and molecular pathways are recognized that induce, or regulate the complex series of interactions required for implantation. In this review, the cellular and molecular interactions that take place during implantation are discussed. [ABSTRACT FROM AUTHOR]

Implantation is a complex process requiring the interaction of the blastocyst, and subsequently the developing embryo with the endometrium. Initially, the detailed cellular interactions implicated in this process were defined. More recently, many signals and molecular pathways are recognized that induce, or regulate the complex series of interactions required for implantation. In this review, the cellular and molecular interactions that take place during implantation are discussed. [ABSTRACT FROM PUBLISHER]

Tumour necrosis factor (TNF)-alpha induced, in a time- and dose-dependent fashion, dyscohesion (cell-cell dissociation) of the endometrial epithelial cells. TNF-alpha impaired the ability of cells to aggregate and to attain compaction. The cell-cell adherent junction is a specialized region of the plasma membrane where cadherin molecules act as adhesion molecules and actin filaments are densely associated with the plasma membrane through a well-developed plasmalemmal undercoat. Dyscohesion induced by TNF-alpha was associated with the disordered expression of cadherin/beta-catenin at the sites of cell-cell contact. In addition, within the time-frame that dyscohesion was induced, TNF-alpha down-regulated the expression of actin mRNA only at 100 ng/ml without modulating the overall amount of actin protein, its beta-isoform or the amount of ribosylated actin. However, TNF-alpha-mediated dyscohesion of epithelial cells was associated with loss of plasmalemmal undercoat as well as intracytoplasmic aggregates of F-actin and a simultaneous increase in G-actin. The effect of cytochalasin-B, which disrupts actin filaments on cell-cell binding, was less pronounced than the effect of TNF-alpha, suggesting that the effect of this cytokine on dyscohesion is not solely dependent on the disassembly of actin filaments. These findings show that the induction of disordered expression of adhesion molecules, as well as disassembly of actin filaments, are implicated in the dyscohesion induced by TNF-alpha. [ABSTRACT FROM AUTHOR]

Endometrium is unique since it is the only tissue that undergoes regular cyclic bleedings. Menstrual shedding is associated with the breakdown of endometrium, including the fragmentation of endometrial glands. To gain insight into the underlying basis of fragmentation of the endometrial epithelium during the menstrual phase, we examined the expression of proteins implicated in epithelial cell-cell binding in human endometria throughout the entire menstrual cycle. Western blotting failed to reveal differences in the relative amount of E-cadherin, alpha- or beta-catenin or actin in the menstrual endometria compared with those in the proliferative or secretory phases. However, specific changes in the expression pattern of these proteins as well as desmoplakin I/II were detected by immunohistochemical staining in epithelial cells of menstrual endometria. Desmoplakin I/II, E-cadherin, alpha- and beta-catenins and beta-actin were localized to intercellular borders as well as the luminal and basal regions of glandular epithelium during the proliferative and secretory phases. Immunoreactivity of E-cadherin and alpha-catenin was confined to epithelial cells, whereas beta-catenin and beta-actin were present in epithelial cells, as well as in stroma and endothelial cells. Binding of F-actin to fluorescein isothiocyanate-labelled phalloidin localized this form of actin to the intercellular borders, and the basal and luminal cytoplasm of epithelial cells in proliferative and secretory endometria. Menstrual shedding was associated with disorganization of the site-specific distribution of desmoplakin I/II, E-cadherin and alpha- and beta-catenins.(ABSTRACT TRUNCATED AT 250 WORDS) [ABSTRACT FROM AUTHOR]

Endometrium is unique since it is the only tissue that undergoes regular cyclic bleedings. Menstrual shedding is associated with the breakdown of endometrium, including the fragmentation of endometrial glands. To gain insight into the underlying basis of fragmentation of the endometrial epithelium during the menstrual phase, we examined the expression of proteins implicated in epithelial cell—cell binding in human endometria throughout the entire menstrual cycle. Western blotting failed to reveal differences in the relative amount of E-cadherin, α- or β-catenin or actin in the menstrual endometria compared with those in the proliferative or secretory phases. However, specific changes in the expression pattern of these proteins as well as desmoplakin I/II were detected by immunohistochemical staining in epithelial cells of menstrual endometria. Desmoplakin I/II, E-cadherin, α- and β-catenins and β-actin were localized to intercellular borders as well as the luminal and basal regions of glandular epithelium during the proliferative and secretory phases. Immunoreactivity of E-cadherin and α-catenin was confined to epithelial cells, whereas β-catenin and β-actin were present in epithelial cells, as well as in stroma and endothelial cells. Binding of F-actin to fluorescein isothiocyanate-labelled phalloidin localized this form of actin to the intercellular borders, and the basal and luminal cytoplasm of epithelial cells in proliferative and secretory endometria. Menstrual shedding was associated with disorganization of the site-specific distribution of desmoplakin I/II, E-cadherin and α- and β-catenins. This included focal patchy expression of these proteins or their total loss from cell—cell binding sites, as well as loss of F-actin in epithelial cells of menstrual endometrial glands. However, these changes were not observed in the basalis region that is not shed during the menstrual phase. These findings suggest that fragmentation of the endometrial glands during the menstrual phase is related to the disorganization and/or loss of the proteins at the adherens and desmosomal junctions, as well as loss of F-actin. [ABSTRACT FROM PUBLISHER]

Tumour necrosis factor (TNF)-α induced, in a time- and dose-dependent fashion, dyscohesion (cell-cell dissociation) of the endometrial epithelial cells. TNF-α impaired the ability of cells to aggregate and to attain compaction. The cell-cell adherent junction is a specialized region of the plasma membrane where cadherin molecules act as adhesion molecules and actin filaments are densely associated with the plasma membrane through a well-developed plasmalemmal undercoat. Dyscohesion induced by TNF-α was associated with the disordered expression of cadherin\β-catenin at the sites of cell-cell contact. In addition, within the time-frame that dyscohesion was induced, TNF-α down-regulated the expression of actin mRNA only at 100 ng/ml without modulating the overall amount of actin protein, its β-isoform or the amount of ribosylated actin. However, TNF-α-mediated dyscohesion of epithelial cells was associated with loss of plasmalemmal undercoat as well as intracytoplasmic aggregates of F-actin and a simultaneous increase in G-actin. The effect of cytochalasin-B, which disrupts actin filaments on cell-cell binding, was less pronounced than the effect of TNF-α, suggesting that the effect of this cytokine on dyscohesion is not solely dependent on the disassembly of actin filaments. These findings show that the induction of disordered expression of adhesion molecules, as well as disassembly of actin filaments, are implicated in the dyscohesion induced by TNF-α. [ABSTRACT FROM PUBLISHER]

Apoptosis in human endometrial epithelium progressively increases from early to late secretory/menstrual phases and remains consistently more prominent in the basalis. It has been suggested that tumour necrosis factor (TNF) alpha secreted during the secretory/menstrual phases plays a role in induction of programmed cell death in these cells. In the present study, we characterized expression of receptors of TNF alpha, Fas antigen and BCL-2 in endometrial cells to gain insight as to whether this type of cell death in endometrium may be related to differential or preferential expression of these proteins at specific phases of the menstrual cycle. In addition, to relate production of TNF alpha to the development of apoptosis, the amount of TNF alpha released by human endometrium was measured. Immunostaining demonstrated that the TNF receptor (TNFr; p55/60)-I, TNFr-II (p75/80) as well as Fas protein were expressed in endometrial epithelium throughout the entire menstrual cycle. This expression was progressively diminished from the basalis towards the upper functionalis. In the proliferative phase, the expression of BCL-2 was prominent in the endometrial glands particularly in those residing in the basalis. This expression became weak as early as the third post-ovulatory day and remained low during the remaining phases of the menstrual cycle. The amount of TNF alpha released by endometrial fragments obtained from various phases of the menstrual cycle was determined. The amount of TNF alpha released into the culture medium by the endometrium was low in the proliferative phase. However, the amount of released TNF alpha progressively increased in the secretory phase and peaked in the menstrual phase. TNFr-I, TNFr-II, Fas, BCL-2 and TNF alpha could be identified by Western blot analysis of proteins extracted from endometrium. Therefore, endometrial epithelium by virtue of expression of receptors of TNF alpha as well as Fas protein is properly poised to respond to ligand signals that regulate apoptosis. Induction of apoptosis in endometrial epithelium and menstrual shedding may be related to loss of the protective effect of BCL-2 as well as to the amount of TNF alpha. [ABSTRACT FROM AUTHOR]

Apoptosis in human endometrial epithelium progressively increases from early to late secretory/menstrual phases and remains consistently more prominent in the basalis. It has been suggested that tumour necrosis factor (TNF) α secreted during the secretory/menstrual phases plays a role in induction of programmed cell death in these cells. In the present study, we characterized expression of receptors of TNFα, Fas antigen and BCL-2 in endometrial cells to gain insight as to whether this type of cell death in endometrium may be related to differential or preferential expression of these proteins at specific phases of the menstrual cycle. In addition, to relate production of TNFa to the development of apoptosis, the amount of TNFα released by human endometrium was measured. Immunostaining demonstrated that the TNF receptor (TNFr; p55/60)-I, TNFr-II (p75/80) as well as Fas protein were expressed in endometrial epithelium throughout the entire menstrual cycle. This expression was progressively diminished from the basalis towards the upper functionalis. In the proliferative phase, the expression of BCL-2 was prominent in the endometrial glands particularly in those residing in the basalis. This expression became weak as early as the third post-ovulatory day and remained low during the remaining phases of the menstrual cycle. The amount of TNFα released by endometrial fragments obtained from various phases of the menstrual cycle was determined. The amount of TNFα released into the culture medium by the endometrium was low in the proliferative phase. However, the amount of released TNFα progressively increased in the secretory phase and peaked in the menstrual phase. TNFr-I, TNFr-II, Fas, BCL-2 and TNFα could be identified by Western blot analysis of proteins extracted from endometrium. Therefore, endometrial epithelium by virtue of expression of receptors of TNFα as well as Fas protein is properly poised to respond to ligand signals that regulate apoptosis. Induction of apoptosis in endometrial epithelium and menstrual shedding may be related to loss of the protective effect of BCL-2 as well as to the amount of TNFα. [ABSTRACT FROM PUBLISHER]

Recently, the demarcating boundaries that allowed separation of the fields of reproductive biology, endocrinology, immunology and neurobiology have faded. The missing link that now ties these disciplines together is the understanding that the language by which cells communicate within these diverse systems is unanimous. This language is the network of products collectively called cytokines. The effect of these factors spans from the hypothalamus to the endometrium and is undoubtedly involved in the maintenance of the delicate balance within the hypothalamic—pituitary—gonadal—endometrial axis. Orchestrated networks of these cytokines also seem to be linked to the steroid hormone signals, an essential feature for maintenance of normal menstrual cycles. Evidence in favour of these emerging concepts is discussed. Major emphasis is placed on interferons, interleukins, tumour necrosis factor, transforming growth factors and colony-stimulating factors. [ABSTRACT FROM PUBLISHER]

Expression of human leukocyte antigen (HLA)-DR molecules and proliferation of epithelium in human endometrium are polarized. We have suggested that the induction of such a polarized micro-environment is T cell and interferon (IFN)-gamma dependent. The present study was designed to demonstrate the induction of such a micro-environment around T cells and around the source of IFN-gamma. Spheroids reminiscent of endometrial glands were formed by allowing three-dimensional aggregation of endometrial epithelial cells of a cloned HLA-DR negative endometrial carcinoma cell line (ECC1) over agarose. Both HLA-DR expression and inhibition of proliferation were found to be directly dependent on the dose of IFN-gamma that was allowed to diffuse in the agarose beneath the spheroids. To show that the interaction of the epithelial cells with activated T cells also induces HLA-DR molecules in a paracrine fashion in the epithelial cells, ECC1 spheroids were co-cultured with increasing numbers of allogeneic peripheral blood T cells for various time-intervals. T cells bound to the ECC1 cells, and became activated as indicated by the expression of interleukin (IL)-2 receptor and HLA-DR molecules. A focal HLA-DR expression became apparent in the ECC1 cells adjacent to the T cells. As the number of T cells added to spheroid cultures was increased, a concomitant increase in the number of HLA-DR positive ECC1 cells occurred and HLA-DR immunoreactivity was enhanced in each cell. There was a corresponding decrease in the proliferation of the ECC1 cells in T cell-ECC1 spheroid co-cultures. Based on these data, we suggest that activation of T cells is associated with the induction of HLA-DR expression and inhibition of proliferation in a paracrine fashion in the epithelial cells and may be responsible for the creation of a polarized micro-environment in vivo. [ABSTRACT FROM AUTHOR]

Recent evidence suggests that diverse endometrial functions may be regulated by cytokines. In this report, the presence of protein and mRNA of cytokines were studied in human endometrium throughout the menstrual cycle. The presence of the interleukin-1 (1L-1)α, interieukin-1 (IL-l)β, interleukin receptor antagonist (TRAP), interleukin-6 (IL-6) and transforming growth factor (TGF)α proteins were demonstrated by immunohistochemical staining. The IL-lα and TGF-α proteins were strongly expressed and IL-1β protein was weakly expressed in all the cells in the stroma as well as epithelial cells. IRAP was markedly expressed in the cells with morphological features of macrophages scattered in the stroma, and the expression of IL-6 protein was predominant in the endometrial epithelium. Diffuse cytoplasmic expression of IL-lα in endometrial epithelium during the pro-liferative phase contrasted markedly with its enhanced luminal expression during the secretory phase of the menstrual cycle. In addition, the presence of the mRNA of these cytokines in endometrium was established throughout the entire menstrual cycle by reverse transcription-polymerase chain reaction (RT-PCR). Abundant expression of cytokines in human endometrium emphasizes the significant roles that cytokines play in cell-cell interactions and in regulating endometrial functions. [ABSTRACT FROM PUBLISHER]

A heterogeneous group of cells interact with each other and with the surrounding matrix to form the complex structure of human endometrium. Since the integrin superfamily of molecules is involved in the cell-cell and cell-matrix interactions, this study was designed to screen, , the cellular distribution of CDW49a-f molecules in human endometrium throughout the menstrual cycle. The integrin molecules were localized by immunohistochemistry using monoclonal antibodies. Glandular epithelium expressed all integrin molecules. With the exception of CDW49d (αβ), surface epithelium also expressed all these molecules. Endothelial cells were positive for all integrin molecules except CDW49a (αβ). Endometrial lymphoid cells were positively immuno-stained for CDW49a, d and e (αβ) and were negative for CDW49b (αβ), CDW49c (αβ) and CDW49f (αβ). Regional differences in the expression of integrin molecules were observed. As compared to the functionalis epithelium, basalis epithelium characteristically exhibited higher expression of CDW49a, d and e. Two integrins in endometrium, CDW49a and d exhibited changes related to the menstrual cycle. CDW49a, which was not expressed in glandular epithelial cells in the proliferative phase, was strongly expressed in these cells after ovulation and its expression was diminished in the late secretory phase. This molecule was not expressed in the stromal cells, however, predecidual cells characteristically expressed this molecule in the late secretory phase. CDW49d was only expressed in the glandular epithelial cells in the mid-proliferative to mid-secretory phases. The endometrial pattern of expression of integrins shows the large array of molecules involved in the cell-cell and cell-matrix interactions in human endometrium and demonstrates the existence of specific microenvironments in this tissue. In addition, the menstrual cycle-related changes in the expression of these molecules are probably related to the cyclic fluctuations in the levels of steroid hormones. [ABSTRACT FROM PUBLISHER]

Examines the regulated expression of cytokines in human endometrium throughout the menstrual cyclelevel of expression of cytokine mRNA during the proliferative and secretory phase. Level of cytokine mRNA expression during gestation; Correlation between habitual abortion and the abnormal expression of interleukin-1beta and interleukin-6 in the mid-secretory phase.

Identifies a novel member of the transforming growth factor (TGF)-beta superfamily and shows that this gene, designated as endometrial bleeding associated factor (ebaf) or TGFbeta4, has a unique expression pattern in human endometrium. Expression of ebaf in endometrium during the late secretory and menstrual phases; Focal expression of TGFbeta4 messenger RNA.