[Show abstract][Hide abstract] ABSTRACT: Classen-Linke I, Moss S, Gröting K, Beier H M, Alfer J & Krusche C A (2012) Histopathology Mammaglobin 1: not only a breast-specific and tumour-specific marker, but also a hormone-responsive endometrial protein Aims: The secretoglobin mammaglobin 1 (MGB1) is strongly expressed in breast tumours, and is therefore used to detect breast cancer metastases, although it has also been detected in other tissues. The aim of this study was to examine MGB1 expression and its hormonal regulation in human endometrium to further investigate the use of MGB1 as a marker molecule. Methods and results: Mammaglobin 1 expression was assessed immunohistochemically in endometrial samples from 60 normal fertile patients throughout the menstrual cycle, in 49 endometriotic tissue samples, in 15 endometrial adenocarcinomas, and in 36 breast carcinomas. In addition, 25 endometrial samples were analysed by western blot and quantitative real-time reverse transcription polymerase chain reaction. To prove hormonal regulation, primary endometrial epithelial cells were cultured with 17β-oestradiol and promegestone. MGB1 was detected in human endometrial tissue, with peak expression during the luteal phase, in 31% of endometriotic samples, in 53% of endometrial adenocarcinomas, and in 64% of breast carcinomas. MGB1 mRNA expression was increased in vitro by hormonal treatment. Conclusions: Our data show that MGB1 expression is not restricted to normal and malignant breast tissue. Besides its documented occurrence in endometriotic and malignant endometrial tissues, MGB1 is also expressed in normal human endometrium, and such expression is controlled by steroid hormones.
[Show abstract][Hide abstract] ABSTRACT: Class I histone deacetylases (HDACs) and acetylases (HATs) are members of transcriptional pre-initiation complexes assembled by steroid hormone receptors. Recently, HDAC inhibitors were shown to enhance differentiation of endometrial fibroblasts and endometrial adenocarcinomas. However, there is only rare information on HDAC and HAT expression in the human endometrium.
HDAC-1, -2, -3 and HAT (PCAF and GCN5) mRNA expression was studied in tissue from premenopausal women undergoing hysterectomy by real-time or semiquantitative RT-PCR. HDAC protein expression was assessed by Western Blot and immunohistochemistry. In endometrial adenocarcinomas (n = 17), HDAC-1 expression was studied by immunohistochemistry.
In the human endometrium, HDAC-1, -2, -3 and PCAF mRNA are expressed without cyclical changes. Western blot analysis demonstrated that HDAC-2 protein expression was slightly, but significantly elevated in the secretory phase (P < 0.01 versus day 5-8), whereas HDAC-1 and -3 protein expression was constitutive throughout the menstrual cycle. By immunohistochemistry, nuclear expression of HDAC proteins was detected in all endometrial cell types. In the case of HDAC-3, immunostaining was significantly reduced in the endometrial surface epithelium on day 6-10 (P < 0.01 versus days 15-18 and 24-28). Compared to normal endometrium, a high proportion of endometrial adenocarcinomas showed impaired HDAC-1 protein expression in the epithelial and stromal compartment.
Class I HDACs and HATs are expressed in the human endometrium throughout the menstrual cycle, suggesting the cyclic endometrium as a potential target for HDAC inhibitors. We hypothesis that alterations of HDAC and/or HAT expression are potentially involved in impaired endometrial differentiation.
Human Reproduction 11/2007; 22(11):2956-66. · 4.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The clinical outcome of treatment of odontogenic cysts differs depending on separate entities. Particular clinical relevance must be attached to the distinction between odontogenic keratocysts, which have an evident tendency to recur, and other odontogenic cysts. The aim of this study was to evaluate cytokeratin (CK) expression patterns as an additional tool for characterization of different cysts as the histomorphologic appearance often is not decisive.
Thirty cases of dentigerous and radicular cysts respectively as well as 15 cases of odontogenic keratocysts were considered. Expression of CK 5/6, 7, 10, 13, 17, 19 and 20 was determined in addition to Ki-67 immunohistochemically.
Expression of CK 17 was discernible in 93.3% of the odontogenic keratocysts, but only in 35.0% of dentigerous and radicular cysts under study (P < 0.001). CK 19 could be detected in 48.3% of dentigerous and radicular cysts, whereas odontogenic keratocysts were completely negative (P < 0.002).
Immunohistochemical detection of CK 17 and 19 seems to be a valuable additional parameter distinguishing between odontogenic keratocysts and other odontogenic--especially dentigerous--cysts which clinically are likely the most significant differential diagnoses in this context. J Oral Pathol Med (2005) 34: 558-64.
Journal of Oral Pathology and Medicine 11/2005; 34(9):558-64. · 2.06 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The most frequent complications of total hip arthroplasty are septic and aseptic wear-induced loosening. A reliable differentiation between septic and aseptic loosening with current diagnostic tools is not possible. Therefore, we examined the diagnostic valency of positron emission tomography (PET) with (18)F-fluorodeoxyglucose (FDG) in cases of septic or aseptic hip arthroplasty loosening compared with conventional triple-phase bone scan (TPBS).
Fifty patients with 70 total hip replacements (symptomatic n=50, asymptomatic n=20) were examined by means of FDG-PET and TPBS to detect septic and aseptic loosening and differentiate between the two. A differentiated algorithm subdivided into categories I-V was developed for FDG-PET. Additionally, standardized uptake values (SUV) were calculated from the lesion with the highest FDG uptake. Interpretations of the TPBS were done according to the criteria described by Wilson. The final diagnosis was based on operative findings including microbiological and histological examinations (n=50), while the remaining asymptomatic arthroplasties (n=20) were integrated into a clinical follow-up (> or =9 months).
Sensitivity/specificity of FDG-PET was 91%/92% (accuracy 91%) compared with 78%/70% (accuracy 74%) for TPBS. A high correlation could be proved between FDG-PET investigation and operative histopathological findings (r(Spear)> or =0.9). No significant differences were found regarding cemented and uncemented implanted hip arthroplasties (p> or =0.05). Calculation of the SUV turned out to be inappropriate as a sole criterion for image interpretation.
FDG-PET is a promising, highly accurate examination method to detect polyethylene and metal wear-induced chronic inflammation followed by periprosthetic osteolysis. In addition, FDG-PET has a significantly higher sensitivity and specificity than TPBS for differentiating between aseptic loosening and infection.
Archives of Orthopaedic and Trauma Surgery 06/2005; 125(5):322-9. · 1.36 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Impaired histone acetylation was recognized to be involved in carcinogenesis. Furthermore, histone deacetylase (HDAC) inhibitors induce differentiation of breast cancer cells and inhibit tumour growth. These results prompted us to study HDAC-1 and -3 expression in breast tumours to establish their potential therapeutic and prognostic significance. HDAC-1 und HDAC-3 protein expression was analyzed immunohistochemically on a tissue microarray (TMA) containing 600 core biopsies from 200 patients. HDAC-1 and -3 expression was correlated to steroid hormone receptor-, Her2/neu- and proliferation status of tumours as well as to overall and disease free survival. Moderate or strong nuclear immunoreactivity for HDAC-1 was observed in 39.8% and for HDAC-3 in 43.9% of breast carcinomas. HDAC-1 and -3 expression correlated significantly with oestrogen and progesterone receptor expression (both p< 0.001). HDAC-1 expression predicted significantly better disease free survival (DFS: p=0.044), in particular, in patients with small tumours of all differentiation types (DFS: p=0.016). Multivariate analysis demonstrated that HDAC-1 is an independent prognostic marker. Our data suggest that evaluation of HDAC-1 protein expression enables a more precise assessment of the prognosis of breast cancer patients. Thus, HDAC-1 expression analysis might be clinically useful to facilitate an individual, risk-directed, adjuvant systemic therapy in breast cancer patients.
Breast Cancer Research and Treatment 04/2005; 90(1):15-23. · 4.47 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The etiology and the pathogenesis of congenital pseudarthrosis of the tibia (CPT) are still unknown. The affected tibia exhibits insufficient mechanical strength and osteogenetic capability. CPT is frequently associated with neurofibromatosis type 1 (NF1; von Recklinghausen's disease); however, both diseases have not yet been linked pathogenetically. This study presents the pathomorphologic findings of CPT under special consideration of NF1. Therefore, samples from patients operated on for CPT (n = 4) with (n = 3) and without (n = 1) neurofibromatosis were investigated by light microscopy, immunohistochemistry, and electron microscopy. The most striking finding in all patients was thickened periosteum with accumulation of nerval cells surrounding small arteries, causing subtotal or complete obliteration. In conclusion, impaired vascularization can result in decreased osteogenic capabilities. The similarity of ultrastructural findings in the abnormal periosteum and in skin neurofibromas of neurofibromatosis patients may indicate a pathogenetic association of both diseases.
Pathology - Research and Practice 02/2005; 201(4):305-12. · 1.21 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The human endometrium prepares for implantation of the blastocyst by reorganization of its whole cellular network. Endometrial stroma cells change their phenotype starting around the 23rd day of the menstrual cycle. These predecidual stroma cells first appear next to spiral arteries, and after implantation these cells further differentiate into decidual stroma cells. The phenotypical changes in these cells during decidualization are characterized by distinct changes in the actin filaments and filament-related proteins such as alpha-actinin. The carboxy-terminal LIM domain protein with a molecular weight of 36 kDa (CLP36) is a cytoskeletal component that has been shown to associate with contractile actin filaments and to bind to alpha-actinin supporting a role for CLP36 in cytoskeletal reorganization and signal transduction by binding to signaling proteins. The expression patterns of CLP36, alpha-actinin and actin were studied in endometrial stroma cells from different stages of the menstrual cycle and in decidual stroma cells from the 6th week of gestation until the end of pregnancy. During the menstrual cycle, CLP36 is only expressed in the luminal and glandular epithelium but not in endometrial stroma cells. During decidualization and throughout pregnancy, a parallel upregulation of CLP36 and smooth muscle actin, an early marker of decidualization in the baboon, was observed in endometrial decidual cells. Since both proteins maintain a high expression level throughout pregnancy, a role of both proteins is suggested in the stabilization of the cytoskeleton of these cells that come into close contact with invading trophoblast cells.
[Show abstract][Hide abstract] ABSTRACT: Embryo implantation and subsequent decidualization, trophoblast invasion and formation of a functional placenta are crucial for establishment and maintenance of pregnancy. Interleukin-11 signalling has been shown to be obligatory for adequate decidualization and trophoblast invasion in mice. Defects in IL-11 signalling in mice result in trophoblast over-invasion and fetal loss. The pathological situation of human tubal pregnancy resembles that of IL-11Ralpha(-/-) mice concerning these symptoms. As our interest is focused on the human early pregnancy, we compared IL-11 expression at the implantation site of ectopic tubal pregnancy (EP) to 1st and 2nd trimester of normal intrauterine pregnancies (IP), and to the normal cycling endometrium. The mRNA expression of IL-11 and IL-11Ralpha was analysed by semiquantitative RT-PCR. Protein expression was detected by western blotting and immunohistochemistry. IL-11Ralpha is expressed constitutively in all tissue specimens analysed. IL-11 is expressed predominantly during follicular and early luteal phase of the menstrual cycle. In IP, IL-11 expression peaks during the 1st trimester and declines from the beginning of the 2nd trimester onwards. In tubal abortions, IL-11 expression is reduced in comparison to vital EP and IP. Cultured primary endometrial and decidual epithelial cells were analysed for hormonal regulation of IL-11 by enzyme-linked immunosorbent assay and RT-PCR. IL-11 is up-regulated by estrogen and down-regulated by progesterone. Overall, our results indicate that in humans, IL-11 signalling is significantly involved in regulation of trophoblast invasion. In the case of tubal abortion, inadequate IL-11 signalling may therefore result in dysregulation of trophoblast invasion.
Molecular Human Reproduction 12/2004; 10(11):783-92. · 4.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: During the first trimester of pregnancy extravillous trophoblast cells (EVT) invade the maternal decidua. Invasion normally is reduced from the second trimester onwards and stops in the inner third of the myometrium. By contrast, in extrauterine tubal pregnancy, trophoblast invasion may even penetrate the tubal wall, which ultimately leads to the rupture of the fallopian tube. Induction of apoptosis of EVT cells, by maternal immune competent cells, may be an important mechanism to limit EVT invasion in uterine pregnancy. Tissue specimens from first and second trimester uterine pregnancy and first trimester tubal pregnancy were analyzed for apoptosis by TUNEL- and M30-staining. By immunohistochemical double labelling, maternal leukocyte subtypes were co-localized to apoptotic cells and in this context, the number of CD56(+)NK cells was analyzed. Our data show that apoptosis is confined to the decidua basalis. Most apoptotic cells are single cytokeratin-positive epithelial cells residing in the stromal compartment. Consequently these cells can only be EVT cells. Maternal leukocytes are not apoptotic. They are located in close contact to apoptotic cells. The number of apoptotic cells in the second trimester (1.8+/-0.7 per cent) is reduced compared to first trimester (5.6+/-0.7 per cent) of uterine pregnancy. In parallel, the number of NK cells declines from first (24.4+/-2.9) to second (12.4+/-1.8) trimester. Furthermore, apoptosis is significantly reduced in ectopic (0.9+/-0.3 per cent) compared to eutopic first trimester pregnancies. Consequently, we suggest that in first trimester uterine pregnancy, induction of EVT cell apoptosis by the maternal immune system is one mechanism to limit EVT invasion. During the second trimester, in parallel to declining numbers of NK cells, the mechanism changes. However, in tubal pregnancy due to differing immunological microenvironments at the ectopic implantation site, apoptosis induction fails, which deleteriously may result in uncontrolled invasion and penetration of the tubal wall.
[Show abstract][Hide abstract] ABSTRACT: The classification of cystic kidney diseases according to the pathologic-anatomic potter classification may be difficult. New molecular genetic findings are important to understand the underlying pathogenesis, but less useful to classify the hereditary diseases. An exact classification of polycystic kidney disease in fetus and children is very important for the human genetic consultation. Therefore, the investigation of pathological anatomy of kidney and liver, as well as the evaluation of additional malformations and family history is necessary. For clinical use the mode of inheritance (autosomal dominant and autosomal recessive) is used to differentiate hereditary polycystic kidney diseases.
Der Pathologe 11/2003; 24(6):410-20. · 0.62 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Die diagnostische Typisierung zystischer Nierenerkrankungen kann im Einzelfall Schwierigkeiten bereiten, da die nach Potter benannten pathologisch-anatomisch definierten Typen oft keine eindeutige Klassifikation erlauben. Die molekulargenetischen Erkenntnisse haben neue Einsichten geschaffen, tragen in der Praxis jedoch nur wenig zur Einteilung bei. Da eine przise Klassifikation vor allem kindlicher, aber auch fetaler Zystennieren fr die humangenetische Beratung betroffener Familien von groer Bedeutung ist, sollte neben einer przisen pathoanatomischen Beschreibung von Vernderungen der Nieren und der Leber, sowie eventuell weiterer Fehlbildungen, immer auch die Familienanamnese bercksichtigt werden. In der klinischen Praxis werden fr die erblichen polyzystischen Nierenerkrankungen die genetischen Begriffe (autosomal-dominant und autosomal-rezessiv) bevorzugt.The classification of cystic kidney diseases according to the pathologic-anatomic potter classification may be difficult. New molecular genetic findings are important to understand the underlying pathogenesis, but less useful to classify the hereditary diseases. An exact classification of polycystic kidney disease in fetus and children is very important for the human genetic consultation. Therefore, the investigation of pathological anatomy of kidney and liver, as well as the evaluation of additional malformations and family history is necessary. For clinical use the mode of inheritance (autosomal dominant and autosomal recessive) is used to differentiate hereditary polycystic kidney diseases.
Der Pathologe 10/2003; 24(6):410-420. · 0.62 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To analyze the impact of cellular proliferative activity and apoptosis, MIB-1 immunopositivity and the apoptotic rate of normal tissue (n = 20), follicular adenoma (n = 30), follicular carcinoma (n = 32), papillary carcinoma (n = 40), Hashimoto thyroiditis (n = 17) and de Quervain thyroiditis (n = 12) was investigated by means of TV-image analysis. Three-micron sections from paraffin-embedded surgical specimens were investigated. Immunohistochemical reactions were performed using an indirect peroxidase method. For determination of apoptosis the in situ DNA nick-end labeling method (TUNEL) was used. The rate of positive cells was determined using the CM-2 TV-image analysis system (Hund, Wetzlar, Federal Republic of Germany). Forty viewing fields (1.94 mm2) were measured with 20:1 objective mangnification. An average of > 4,400 cells were assessed in each case. The mean MIB-1 immunopositivity was higher in follicular carcinoma (average, 2.30%) than in de Quervain thyroiditis (1.48%), papillary carcinoma (1.26%), Hashimoto thyroiditis (0.97%), follicular adenoma (0.58%) and normal thyroid tissue (0.14%). The apoptotic rates were higher in Hashimoto thyroiditis (4.54%) and de Quervain thyroiditis (3.55%) than in thyroid tumors (0.31%-0.49%) and normal thyroid tissue (0.10%). Calculating the ratio of MIB-1 expression and apoptotic rate, thyroid tumors (12.1-14.6) revealed higher values than normal thyroid tissue (3.6), indicating cell gain. In Hashimoto thyroiditis (1.7) and de Quervain thyroiditis (0.7) the ratio was lower than in normal tissue indicating cell loss. MIB-1 immunohistometry and apoptotic rate quantification present new insights into the proliferative and apoptotic potential of thyroid lesions based on a high number of cells investigated per case. The impact of both methods could be used for the interpretation of diagnostically difficult and inconclusive fine-needle aspirates. However, as there was an overlap of the single values, the results have to be interpreted carefully for diagnostic purposes.
Anticancer research 01/2003; 23(5b):4269-75. · 1.71 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Biologisch ist die Reifung des Endometriums in jedem Zyklus darauf gerichtet, einer Blastozyste günstige Bedingungen für die
Implantation zu bieten. Infertilität und frühes Ende einer Schwangerschaft beruhen nicht selten auf einer unvollkommenen Reifung
des Endometriums. Bereits eine mangelhafte Rezeptivität des Endometriums führt zum Scheitern der Implantation. Um die endometriale
Ursache einer Infertilität zu erkennen, werden die normalen zyklischen Veränderungen des Endometriums studiert. Morphologische
Veränderungen bilden bis heute in diesem Bemühen der Gynäkologen noch die Grundlage der klassischen histologischen Diagnostik
nach Noyes et al. (1950). Da jedoch evident ist, dass molekulare Defizite des Endometriums sich nicht unmittelbar histologisch
erfassen lassen, werden neue Methoden und signifikante Parameter gesucht, die diese Art funktioneller Defekte diagnostisch
erfassen lassen. Histochemische Lokalisierungen von definierten Proteinen, von Adhäsionsmolekülen und Zytokinen, erlauben
bereits eine detailliertere und aussagekräftigere molekulare Analyse als es die klassische Morphologie ermöglicht. Überzeugende
Argumente sprechen inzwischen dafür, mit einfachen biochemischen Analysen die Proteine des Uterussekrets als weitergehende
signifikante Parameter zu nutzen. Die elektrophoretische Auftrennung des Uterussekrets ergibt typische Proteinmuster, welche
die zyklischen Funktionsphasen des Endometriums darstellen. Das adäquate Proteinmuster der Lutealphase ist als das Produkt
eines rezeptiven Endometriums zu betrachten und repräsentiert das für diese molekulare Analytik charakteristische „Implantationsfenster“.
Es wird bereits 2 Tage post ovulationem exprimiert und bei ungestörtem Zyklusverlauf 8 Tage aufrechterhalten (15.–24. Zyklustag).
The biological aim of the differentiation and maturation of endometrial tissue compartments during any menstrual cycle is
the achievement of suitable conditions for blastocyst implantation and the establishment of pregnancy. Infertility and early
embryonic loss are frequently caused by insufficient endometrial differentiation. Even any incomplete receptivity stage of
the luteal phase endometrium will prevent attachment and implantation. We have studied the physiological changes throughout
an endometrial cycle to elucidate causes of endometrial insufficiencies leading to subfertility or infertility. Up to now,
the histological changes described by Noyes et al. are understood as classical diagnostic approaches. However, evidence is
accumulating that molecular deficits of endometrial differentiation are by no means detectable histologically, and consequently
require research on new diagnostic methods and parameters. There are histochemical localizations of specific protein molecules,
adhesion molecules and cytokinesthat permit far more detailed and significant molecular analyses than any classical morphological
means could yield. Moreover, there are convincing arguments for using further biochemical assessments on proteins of the uterine
secretions as specific diagnostic parameters. The electrophoretical resolution presents typical protein patterns, which in
turn can be interpreted as characteristic reflections of the functional phases of the endometrial cycle. That which is demonstrated
as the so-called adequate luteal phase protein pattern is clearly the product of the receptive endometrium, reflecting the “implantation window”. This is already established two
days after ovulation and usually persists eight further days if the endometrial cycle is undisturbed (15th to 24th day of the cycle).
[Show abstract][Hide abstract] ABSTRACT: Leptin and its receptor are involved in endocrine and paracrine regulation of metabolism, obesity and reproduction. Here, we describe the detection of the functional long isoform receptor of leptin in human endometrium. The leptin receptor protein was shown to be expressed in glandular and luminal epithelium and is periodically regulated throughout the menstrual cycle, demonstrating main expression in follicular and mid-luteal phase. In contrast, leptin receptor mRNA is detectable by reverse transcription-polymerase chain reaction (RT-PCR) as a constitutive component. Since RT-PCR analyses showed that leptin is not expressed in this tissue, the present study suggests that the human endometrium is a novel target for leptin. Therefore, we investigated 11 subfertile patients who underwent two biopsies in one menstrual cycle. The patients presented with a repetitive endometrial maturation defect, but showed adequate serum hormone concentrations and normal steroid hormone receptor expression and down-regulation in the endometrium. These patients were, however, deficient for expression of the functional leptin receptor. These analyses provide evidence that the lack of the leptin receptor in an ovulatory cycle may contribute to subfertility by a yet undefined 'endometrial factor'.
Molecular Human Reproduction 08/2000; 6(7):595-601. · 4.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Endogenous nitric oxide (NO) can be synthesized by endothelial cells and can act as a potent vasodilator. We investigated endothelial nitric oxide synthase (eNOS), one of the three different enzymes responsible for the synthesis of NO by immunohistochemical methods throughout the menstrual cycle on 34 endometrial samples and compared its detection with the von Willebrand Factor (vWF) as a reliable marker molecule of the endothelium on serial sections. Immunoreactivity for eNOS was clearly localized in various types of arterial and venous endothelial cells as well as in capillaries. In addition, in some samples there was a positive staining in endometrial glandular epithelium. There was no staining in endometrial fibroblasts or in myometrial smooth muscle cells. Whereas the endothelium was constantly stained by the monoclonal antibody against vWF, eNOS was not always expressed in the endothelial lining of the vessels during the menstrual cycle. The number of vessels positively stained for eNOS increased gradually during the proliferation phase and most of the vessels were positive in the early secretory phase. These results suggest that its markedly increased expression during the early secretory phase of the menstrual cycle indicates a physiological significance.
Molecular Human Reproduction 03/2000; 6(2):185-90. · 4.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Ziel der Untersuchungen war die Identifizierung von sogenannten Markermolekülen für ein rezeptives und somit implantationsbereites Endometrium. Hierbei wurden fertile und subfertile Patientinnen untersucht. Sonographisch war das Endometrium der subfertilen Patientinnen während des gesamten ovulatorischen Zyklus < 7mm. Wie tierexperimentelle molekularbiologische Studien zeigten, führen drei der ausgesuchten Faktoren (LIF, Leptin, ß3-Integrin), wenn sie fehlen oder verändert sind zur Infertilität. Folgende Marker wurden immunhistochemisch analysiert: (1.) die Zytokine VEGF und LIF, (2.) das Hormon Leptin und sein Rezeptor und (3.) das ß3-Integrin. Die Analyse der oben genannten Faktoren innerhalb des Menstruationszyklus fertiler Patientinnen zeigt eine Expression dieser Moleküle überwiegend in der mittleren Lutealphase des Zyklus. Dies weist daraufhin, daß auch beim Menschen diese für die erfolgreiche Reproduktion notwendig sind, und ihre Expression daher als Marker für die endometriale Rezeptivität genutzt werden könnte. Den untersuchten subfertilen Patientinnen fehlten diese Marker nahezu vollständig. Die fehlende oder inadäquate Exprimierung der hier getesteten Markermoleküle könnte durch die additive Wirkung von Funktionsstörungen für Subfertilität oder Infertilität verantwortlich sein. Eine wesentliche Gemeinsamkeit der unterschiedlichen Marker ist ihre Einbindung in den zellbiologischen Prozess der Angiogenese. Ihre fehlende Expression im Endometrium der subfertilen Patientinnen könnte auf molekularer Ebene den defizienten Aufbau dieser Endometrien erklären, der seinen wesentlichen Ursprung in einer insuffizienten Angiogenese zu haben scheint. Durch diese Arbeit konnte, bei dem bisher nicht näher definierten endometrialen Faktor, erstmalig eine mögliche Störung der Angiogenese als Ursache für Subfertilität aufgezeigt werden. Aim of this study was to identify marker molecules for a receptive endometrium. We investigated endometrial biopsies of fertile and subfertile patients. The endometrium of subfertile patients was < 7mm for the whole ovulatory cycle. As animal models and molecular investigations demostrated, three of our selected markers (LIF, Leptin, ß3-Integrin) are leading, when absent or mutated, to infertility. The following markers were investigated by immunohistochemistry: (1.) the cytokines VEGF and LIF, (2.) the hormone leptin and its receptor and (3.) the ß3-integrin. The analyses of these markers during the menstrual cycle of fertile patients showed an expression in the endometrium during the midluteal phase. These observations indicate a significant role for a successful reproduction and we postulate that their expression can be used for the assessment of endometrial receptivity. Subfertile patients were almost negative for these markers. The absence or inadequate expression of the tested molecules might be responsible for subfertility or infertility. The fundamental identity of these markers is their involvement in angiogenesis. Their absence in the endometrium of subfertile patients may contribute to a deficient transformation in this tissue as an indicator for insufficient angiogenesis. The present study enabled us to define the "endometrial factor" on a molecular level, in particular by a deficient angiogenesis as a reason for subfertility.
[Show abstract][Hide abstract] ABSTRACT: Uteroglobin is a progesterone binding protein, a member of the antiflammin gene family and possibly a novel cytokine. Initially, uteroglobin was identified as the major protein of rabbit uterine secretion during the phase of preimplantation. Counterparts of the rabbit uteroglobin or its gene are described in rat, mouse, hamster, hare, pig, horse and human. While uteroglobin appears as one of the most extensively studied proteins, particularly its physico-chemical properties, including its crystal structure and its gene, the true physiological role of this protein still remains to be unravelled. Essential to understanding the significance of human uteroglobin in reproductive organs, particularly in the endometrium, is a knowledge of the spatial and chronological expression of this secretory protein. Our studies on 115 volunteers combined reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry and quantitative assessment by an enzyme-linked immunosorbent assay for uteroglobin. The expression, localization and release of uteroglobulin in the human endometrium are presented. Secretory uteroglobin is found in endometrial tissue homogenates in highest levels of expression during the early luteal phase (days 15-19, 340 pg/mg total protein). In turn, uteroglobin is released into the uterine lumen in peak amounts during the receptive phase of the menstrual cycle (mid-luteal phase, days 20-23, secretion level 833.4 pg/mg total protein). Our immunohistochemical studies match with these results, as uteroglobin is located during the early and mid-luteal phase in the apical compartments of endometrial gland cells. These observations strongly suggest an involvement of uteroglobin in endometrial preparations for implantation.
Molecular Human Reproduction 01/2000; 5(12):1155-61. · 4.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The influences of the synthetic progestin, medroxyprogesterone acetate (MPA), the progesterone receptor modulator J867, and the antagonist ZK137316 were studied in vitro on isolated endometrial epithelial cells, as well as endometrial fibroblasts. We evaluated the expression of estrogen receptor α (ER) and the progesterone receptor (PR) by RT-PCR. ER and PR were strongly expressed in the fibroblasts and epithelial cells under treatment with 10−8 M 17β-estradiol (E2). Treatment with 10−6 M J867 or ZK137316 upregulated the PR expression as did E2, in contrast to treatment with 10−6 M MPA, which caused a downregulation of PR in epithelial cells, but not in fibroblasts. In addition, the vascular endothelial growth factor (VEGF) release into the cell culture medium was analyzed by a VEGF-ELISA. VEGF which plays an important role in angiogenesis, is regulated by steroid hormones as well as hypoxia. E2 stimulates VEGF release into the medium in both cell types. MPA reduces VEGF release significantly in the fibroblast cell culture, but increases it in the epithelial cell culture. ZK137316, in the presence or absence of E2, reduces VEGF release in fibroblast cell culture. J867 increases the VEGF production in fibroblasts only in the presence of E2. Both compounds show no significant effects, compared to E2, in epithelial cell culture. The different results for the epithelial cells and fibroblasts indicate that the pharmacological effects of PR modulators (PRMs) and progesterone antagonists (PAs) may be cell specific and depend on the presence or absence of partial progestagenic agonistic activities. This observation opens up new perspectives for various clinical applications.
[Show abstract][Hide abstract] ABSTRACT: An established cell culture system of isolated human endometrial stromal and epithelial cells has been used to study the effects of oestrogen and progesterone, as well as their antagonists, upon endometrial cells. Normal hormonal regulation in vivo was investigated simultaneously in endometrial tissue samples taken at different phases of the menstrual cycle. Several marker molecules analysed by immunohistochemistry appeared to depend strongly on endocrine regulation and could be traced in culture. Immunohistochemically, basic parameters of cell biology were identified in vitro, e.g. cell proliferation (Ki-67), adhesion molecules (beta3 integrin) and paracrine factors (leukaemia inhibitory factor). The most reliable parameters to assess hormonal influences were oestrogen and progesterone receptor molecules. Immunohistochemical localization could be improved by molecular biological analysis using RT-PCR. In the presence of oestrogen, a significant expression of hormone receptors was also shown by RT-PCR, and withdrawal of oestrogens and addition of gestagen, i.e. medroxyprogesterone acetate, caused receptor downregulation. Addition of the anti-oestrogen ICI 182.780 to cell-culture medium significantly decreased the synthesis of progesterone receptors.
Human Reproduction Update 01/1998; 4(5):539-49. · 8.85 Impact Factor