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Biomedical subjects

I Mylonas

Publications and source records attributed to I Mylonas.

At least 19 recordsLinked to original sources
PubMed2007

Steroid receptors ERalpha, ERbeta, PR-A and PR-B are differentially expressed in normal and atrophic human endometrium.

OBJECTIVE: The endometrium expresses estrogen (ER) and progesterone receptor (PR), which are related to autocrine and paracrine processes that respond to estrogen and progesterone. Therefore, the aim of this study was to evaluate the distribution pattern of ERalpha, ERbeta, PR-A and PR-B with monoclonal antibodies in normal human endometrial tissue. STUDY DESIGN: Human endometrial tissue was obtained from 84 premenopausal and 11 postmenopausal patients and immunohistochemically analysed with monoclonal antibodies against ERalpha, ERbeta, PR-A and PR-B. RESULTS: ERalpha, PR-A and PR-B declined significantly (p<0.001, p<0.05, p<0.05 respectively) in glandular epithelium from proliferative to late secretory phase. The ERbeta immunohistochemical reaction showed a similar significant declining pattern (p<0.05), although the staining intensity was lower than that of ERalpha. While ERalpha, ERbeta and PR-B decrease significantly in atrophic endometrial tissue compared to proliferative endometrium, a significant up-regulation of PR-A was observed compared to late secretory phase (p<0.05). CONCLUSION: ERalpha, ERbeta, PR-A and PR-B were expressed in normal human endometrium with a cyclical variation during the menstrual cycle. In normal postmenopausal endometrial tissue, a down-regulation of ERalpha, ERbeta and PR-B occurs with a subsequent higher expression of PR-A. These results show the presence of steroid receptors in human epithelium, indicating that these cells respond to estrogen and progesterone, thus playing a significant role in endometrial physiology.

Atrophy↗
PubMed2006

Quality of life among breast cancer patients undergoing autologous breast reconstruction versus breast conserving therapy.

PURPOSE: Besides the quality of the aesthetic results, the quality of life after surgery is one of the most important criteria when reviewing different operation methods, especially in oncologic diseases. This study was performed to evaluate the difference in the health-related quality of life following breast conserving surgery and autologous breast reconstruction after mastectomy. PATIENTS AND METHODS: Hundred and forty-four breast cancer patients were included in this study. Sixty seven patients underwent breast conserving surgery followed by radiotherapy. In 77 patients a mastectomy was performed with immediate or late reconstruction. To evaluate the health-related quality of life we used the SF-36 self-administered questionnaire. RESULTS: A significant difference was found in quality of life in the subscale "physical functioning" showing better results in the breast reconstruction group (P = 0.01). No significant difference was found in the other subscales, but there was a tendency to a better "emotional role" among the breast reconstruction patients. CONCLUSION: Our study demonstrated that autologous tissue breast reconstruction in breast cancer patients did not affect adversely the health-related quality of life compared to breast conserving therapy when the quality of life is assessed by the standardized questionnaire SF-36. In particular, the physical function is not reported to be significantly influenced negatively by the more extensive surgical therapy.

Adult↗
PubMed2006

Expression of inhibin/activin subunits alpha (-alpha), beta A (-beta (A)) and beta B (-beta (B)) in placental tissue of normal and intrauterine growth restricted (IUGR) pregnancies.

During human pregnancy the placenta produces a variety of proteins like steroid hormones and their receptors that are responsible for the establishment and ongoing of the feto-placental unit. Inhibins are dimeric glycoproteins, composed of an alpha-subunit and one of two possible beta-subunits (beta (A) or beta (B)). Aims of the present study were the determination of the frequency and tissue distribution patterns of the inhibin/activin subunits in human placental tissue of normal pregnancies and pregnancies complicated with fetal growth restriction (IUGR). Slides of paraffin embedded placental tissue were obtained after delivery from patients diagnosed with IUGR (n = 6) and normal term placentas (n = 8). Tissue samples were fixed and incubated with monoclonal antibodies inhibin/activin-subunits -alpha, -beta (A), -beta (B). Intensity of immunohistochemical reaction on the slides was analysed using a semi-quantitative score and statistical analysis was performed (P<0.05). A significant lower expression of the inhibin-alpha subunit in IUGR extravillous trophoblast compared to normal pregnancies was observed, while the inhibin-alpha immunostaining was significantly upregulated in syncytiotrophoblast. Additionally, a significant down-regulation of inhibin-beta (B) subunit in extravillous trophoblast cells in IUGR syncytiotrophoblast cells was demonstrated. A co-localisation of inhibin-alpha and the beta-subunits was also observed, suggesting a production and secretion of intact inhibin A and inhibin B. Although the precise role of these inhibin/activin subunits in human placenta and IUGR pregnancies is still unclear, they could be involved in autocrine/paracrine signalling, contributing to several aspects like angiogenesis and tissue remodelling.

Activins↗
PubMed2006

Development and characterization of monoclonal antibodies for the immunohistochemical detection of glycodelin A in decidual, endometrial and gynaecological tumour tissues.

AIMS: Glycodelin is a glycoprotein with a molecular weight of 28 kDa. Unusual LacdiNAc structures have been identified on glycodelin A, isolated from amniotic fluid. Three major functions of this glycoprotein have been identified. Glycodelin is an immunosuppressive molecule, a marker of morphological differentiation, and a contraceptive. Because no monoclonal antibodies for glycodelin A are commercially available, our aim was to develop and characterize three monoclonal antibodies against this glycoprotein. METHODS AND RESULTS: Glycodelin A was purified from amniotic fluid by three chromatographic steps and its purity was checked by SDS-PAGE. Antibodies were generated from immunized BALB/c mice. Three IgG1 monoclonal antibodies detecting glycodelin A were cloned. All three antibodies recognized carbohydrate structures of glycodelin A and did not cross-react with glycodelin S. They are applicable to immunohistochemistry (frozen and paraffin sections), ELISA and Western blots. CONCLUSION: The new antibodies can be used for the detection of glycodelin A in frozen and paraffin-embedded decidual and endometrial tissue. One antibody (A87-B/D2) can be used for the detection of glycodelin in endometrial and ovarian tumour tissues. Because glycodelin A is a major secretory endometrial product during the luteal phase, in early pregnancy and in gynaecological tumours, the new antibodies are, potentially, valuable tools for the study of endometrial development and tumour progression.

Amniotic Fluid↗
PubMed2006

Expression of inhibin/activin subunits alpha (-alpha), betaA (-betaA), and betaB (-betaB) in placental tissue of normal, preeclamptic, and HELLP pregnancies.

During human pregnancy the placenta produces a variety of proteins for the establishment of the fetoplacental unit, including inhibins and activins. Inhibins are dimeric glycoproteins, composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB). Aims of the present study were (a) the determination of the frequency and tissue distribution patterns of the inhibin/activin subunits in human placental tissue of normal pregnancies and pregnancies complicated with preeclampsia and HELLP syndrome (hemolysis, elevated liver enzymes, low platelets) and (b) the assessment of a combined expression of inhibin-alpha- and both beta-subunits (betaA-and betaB-subunits) using double immunofluorescence technique. A significant lower expression of the inhibin-alpha subunit in preeclamptic and HELLP placental tissue compared to normal pregnancies was observed, while the inhibin-alpha immunostaining was significantly upregulated in syncytotrophoblast. Additionally, we demonstrated a significant down-regulation of inhibin-betaB subunit in extravillous trophoblast cells between normal and preeclamptic compared to HELLP placental tissue, while inhibin-betaA-subunit was significantly higher in preeclamptic syncytotrophoblast cells. A colocalization of inhibin-alpha and the beta-subunits could be demonstrated, suggesting a production and secretion of intact inhibin A and inhibin B. Therefore, inhibin A and activin A might be useful markers in preeclampsia. Valuable parameters in HELLP syndrome could be inhibin A, rather than inhibin B, and activin B. Furthermore, the lower betaB-subunit production in extravillous trophoblast cells demonstrates that this subunit might have an important role in the pathogenesis of HELLP syndrome. Additionally, the higher production of the betaA-subunit in syncytotrophoblast cells suggest a higher production of activin A rather than inhibin A in preeclampsia that might be utilized as a marker of placental function.

Adult↗
PubMed2005

Expression of glycodelin A in decidual tissue of preeclamptic, HELLP and intrauterine growth-restricted pregnancies.

During pregnancy, the placenta produces a variety of proteins that are responsible for the establishment of the foeto-maternal tolerance and circulation. The aim of this study was to investigate the expression of glycodelin A (formerly named PP14) in decidual tissue of placentas with intrauterine growth restriction (IUGR), preeclamptic patients, hemolysis, elevated liver, low-platelet (HELLP) patients and normal decidual tissue. Slides of paraffin-embedded decidual tissue of patients with IUGR, preeclamptic patients, HELLP patients and normal-term placentas were incubated with either polyclonal or monoclonal antibodies against glycodelin A. Staining reaction was performed with the ABC reagent. Intensity of immunohistochemical reaction on the slides was analysed using a semi-quantitative score. In addition, expression of glycodelin mRNA was analysed by in situ hybridisation. Expression of glycodelin A was significantly reduced in decidual cells of placentas with IUGR and HELLP, as investigated with both monoclonal and polyclonal antibodies and in situ hybridisation. However, preeclamptic decidual tissue showed no significantly different expression of intensity of glycodelin mRNA compared with normal placental tissue controls. A reduced expression of glycodelin A by decidual cells seems to be related to IUGR and HELLP. Therefore, glycodelin A might play an important role in the pathogeneses of these diseases.

Adult↗
PubMed2005

[Stimulation of HCG, estrogen and progesterone production in isolated trophoblast cells by glycodelin A or its N-glycans].

UNLABELLED: The immunosuppressive protein glycodelin A (GdA) is secreted by decidual tissue in high rates in the first trimester of pregnancy. GdA forms about 10 % of the total protein amount released by the decidua in the first trimester of pregnancy and has a unique glycosylation including fucosylated LacdiNAc structures. The purpose of this study was to investigate the effect of glycodelin A and its N-glycans on hCG, estrogen and progesterone release by cytotrophoblasts in vitro. Trophoblast cells were isolated from human term placenta by fragmentation of villous tissue. Trophoblast cells fuse in vitro to syncytiotrophoblast cells. Trophoblast cells were incubated with GdA and various glycans of GdA. The production of hCG and progesterone was measured after 24 and 48 hours. The production of hCG, estrogen and progesterone was increased in GdA and glycan-treated cell cultures as compared to untreated trophoblast cell cultures. CONCLUSION: HCG, estrogen and progesterone are markers for the differentiation process of cytotrophoblast cells. The results suggest that GdA with its distinct glycosylation modulates the differentiation of trophoblasts.

Cell Differentiation↗
PubMed2004

Glycodelin A and differentiation of first trimester trophoblast cells in vitro.

AIM: The glycoprotein, glycodelin A (GdA) is a main product of the maternal decidua in the first trimester of pregnancy and is secreted into the amniotic fluid. The purpose of this study was to investigate the effect of GdA on secretion and surface markers of isolated first trimester trophoblasts in vitro. METHODS: Cytotrophoblasts were prepared from human first trimester placentae and incubated with varying concentrations of GdA or transfected separately with the expression plasmid of GdA. Supernatants were assayed for human chorionic gonadotropin (hCG) protein concentrations. Expression of human placental lactogen (hPL), mucin 1 (MUC1) and the Thomsen-Friedenreich (TF) epitope was analysed in stimulated trophoblast cells and in unstimulated controls by immunocytochemistry. RESULTS: Glycodelin A induced a reduced expression of hPL compared with unstimulated controls. Expression of MUC1 was not affected by GdA. Freshly isolated trophoblast cells showed no TF expression but became positive for this antigen after 96 h of cultivation. GdA-stimulated trophoblast cells inhibited TF expression after 96 h of cultivation. GdA plasmids induced a significantly higher hCG production in transfected cells than in cells transfected with the empty plasmid. CONCLUSIONS: The results obtained in this study suggest that GdA is involved in the differentiation of trophoblast cells. The treatment of GdA plasmid transfected trophoblast cells stimulated hCG production in isolated trophoblast cells and inhibited hPL and TF expression, suggesting a functional link between hCG and GdA.

Adult↗
PubMed2004

[Investigations on regulation of HCG by cortisol (prednisolon) in trophoblast cells in vitro].

OBJECTIVE: Trophoblast cells synthesize a variety of hormones, in which hCG plays a major role. On the strength of special enzymes they are capable of catalyzing the reaction cortisol <--> cortisone. In vitro experiments showed the influence on ACTH- and cortisol secretion by CRH, ACTH and prednisolon. In this study we describe the influence of cortisol (prednisolon) on hCG production of trophoblast cells in vitro. MATERIAL AND METHODS: Trophoblast cells were prepared from human term placentae by standard trypsin-DNAse dispersion of villous tissue followed by a percoll gradient centrifugation step. After adjusting the cell suspension to a defined cell concentration of 1 x 10 (6) cells/ml cells were cultivated. The addition of prednisolon followed every eight hours. The samples were collected after 24 hours for a total of 96 hours also from unstimulated cultures. Culture supernatants were assayed for hCG by enzyme-immunometric methods. RESULTS: The addition of prednisolon (50 microg/ml) stimulates the concentration of hCG in a time-depending manner. CONCLUSIONS: The trophoblast cell shows an increase in the concentration of hCG after stimulation with cortisol. For the first time an influence of cortisol (prednisolon) on hCG production could be demonstrated in cultured trophoblast cells.

Chorionic Gonadotropin↗
PubMed2003

Immunohistochemical expression of inhibin-alpha in human endometrium and the in vitro secretion of inhibin, estradiol and cortisol in cultured human endometrial glandular cells.

BACKGROUND: Inhibins are multipotent dimeric glycoproteins, composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB). Aims of this study were (a): the immunohistochemical characterisation of normal human endometrium for the inhibin-alpha subunit; (b) the assessment of the secretion and metabolism of inhibin, E2 and cortisol; (c) the evaluation of any relationship between these three substances in cell culture medium of isolated and cultivated normal human endometrial glandular cells. MATERIALS AND METHODS: Samples of human endometrium were obtained from 34 premenopausal patients. Nineteen endometrial specimen (proliferative [PP] n=8; early secretory [ES] n=7; late secretory phase [LS] n=4) were brought into cell culture. Fifteen endometrial specimen (PP n=5; ES n=5; LS n=5) were paraffin-fixed and used for the immunohistochemical analysis for inhibin-alpha. Stromal and epithelial cells were separated by collagenase digestions, filtrations, sedimentations and Ficoll-gradient centrifugation. E2 and cortisol were measured with radioimmunoassay (RIA) and inhibin with enzyme-immunoassay (EIA). Statistical analysis was performed with the non-parametric Mann-Whitney rank-sum test and linear regression analysis. RESULTS: Inhibin-alpha showed a weak (positive) expression during proliferative phase, which increased significantly as the menstrual cycle continued. In secretory glands the mean inhibin concentration was higher than that from proliferative samples. A significant correlation was observed between inhibin and E2 (p<0.001) as well as cortisol and inhibin (p<0.0001) in glands from proliferative phase. Between inhibin and E2 (p<0.05) as well as inhibin and cortisol (p<0.002) a significant correlation in early secretory glands was also noted. In late secretory phase inhibin and E2 (r2=0.78650; p<0.0001), inhibin and cortisol (r2=0.58326; p<0.001) and E2 and cortisol (r2=0.52880; p<0.001) showed a significant correlation. DISCUSSION: In conclusion, we found a cyclical expression of inhibin-alpha subunit in the endometrium demonstrated by immunohistochemical means. A higher in vitro secretion of inhibin from secretory glands was also observed. In addition, a significant correlation between inhibin with E2 and cortisol in PP and ES glands and a significant correlation between inhibin, E2 and cortisol in LS glands could also be demonstrated. We conclude that inhibin can be associated with E2 and cortisol metabolism, playing an important role in paracrine/autocrine mechanisms in the endometrium and possibly exerting its function through cortisol and E2. The cortisol concentration also correlates with E2, suggesting a link between these steroids in the endometrial function. The correlation of inhibin, E2 and cortisol suggest complex autocrine/ paracrine mechanisms in human endometrial glands, modulated and controlled by all these three substances.

Adult↗
PubMed2003

Unilateral ovarian agenesis: report of three cases and review of the literature.

INTRODUCTION: Unilateral ovarian agenesis is a rare event and only a few case have been reported. CASE REPORT: We present three additional cases, where patients presented with diffuse lower abdominal pain. During laparoscopy, an unilateral ovarian agenesis was observed in the three cases. DISCUSSION: There are two possible explanations of a unilateral ovarian absence, involving an asymptomatic adnexal torsion or congenital absence. Unknown environmental factors or genetic predisposition could contribute to this kind of anomaly.

Adult↗
PubMed2003

[Reduction of the risk of breast cancer].

Aspects of nutrition and lifestyle may be largely responsible for the development of common cancers in Western countries, as indicated by the large differences in breast cancer rates between countries, the striking changes in these rates among migrating populations, and the rapid changes over time within countries. The better informed and increasingly health-conscious population of the present day are intensively seeking to identify and eliminate these putative carcinogenic risk factors and to exploit the preventive effects that have been attributed to certain dietary components. Nutrition and 'lifestyle' may exert its carcinogenic effects indirectly by cell stimulations (alcohol, hormone replacement therapy), inhibition of DNA-repair mechanisms (lack of vitamins), effecting estrogen metabolism (phytoestrogen), or as promotors to enhance growth of tumors (body mass index). Some 'substances' may act as a carcinogenic itself, e. g. aromatic hydrocarbons in tobacco or increased polycyclic aromatic hydrocarbons in well done meat. Individual differences in the effects of nutritional factors on mammary epithelia could be caused by genetic polymorphisms. In this critical review, we focus on current data regarding the effect of nutrition and lifestyle, on the risk of developing breast cancer. A health lifestyle, consisting of "healthy diet", physical activity, renunciation of stimulants, is recommended from childhood throughout life.

Body Mass Index↗
PubMed2003

[Investigations on isolation, purification and cultivation of human endometrial cells and on the in vitro inhibin expression in glandular epithelial cells].

The separate in vitro cultivation of isolated and purified human endometrial glands and stromal cells seems to be the most attractive experimental way of studying the endometrial function on cellular level. In this paper a new method has been described to establish monolayer cultures of isolated endometrial stromal and epithelial cell populations. After a first collagenase digestion, stromal and epithelial cells were separated by filtration. The glandular epithelial cells were further purified with two collagenase digestion steps, filtration, a differential sedimentation at unity gravity and a Ficoll gradient centrifugation. Stromal cells were isolated with the use of erylyse-buffer, filtration and differential sedimentation at unity gravity. A significant higher inhibin production was observed during late secretory compared to proliferative and early secretory phase. Therefore, glandular epithelial cells maintain in vitro their initial differentiation. The higher inhibin concentration during secretory phase implicates a substantial role in endometrial function and maturation. Therefore, inhibin could be used as a marker of endometrial differentiation. Experiments on isolated glandular epithelial cells should be performed within two weeks. The method described here allows the propagation in vitro of separate endometrium cell types which can be used to study endometrial function as well as implantation mechanisms.

Cell Culture Techniques↗
PubMed2003

[Investigations on regulation of CRH, ACTH and cortisol in trophoblast cells in vitro].

OBJECTIVE: Trophoblast cells synthesise CRH and ACTH, which are peptide hormones. On the strength of special enzymes they are capable of catalyzing the reaction cortisol <--> cortisone. In vitro experiments should give a proof of influence to ACTH- and cortisol secretion by CRH, ACTH and prednisolon. The basal rate of cortisol secretion was examined in a long term experiment. MATERIAL AND METHODS: Trophoblast cells were prepared from human term placentae by standard trypsin-DNAse dispersion of villous tissue followed by a percoll gradient centrifugation step. After adjusting the cell suspension to a defined cell concentration of 1 x 10(6) cells/ml the cells were cultivated. The addition of CRH, ACTH or prednisolon followed every eight hours. The samples collected 20 or 30 minutes later, also from unstimulated cultures, were assayed for ACTH and cortisol by enzyme-immunometric methods. RESULTS: The concentration of cortisol shows a rhythmical course in long term cell cultures. The addition of CRH (500 ng/ml, 1 microg/ml) stimulates the concentration of ACTH- and cortisol in a time-depending manner. The addition of ACTH (500 ng/ml-2 microg/ml) stimulates the concentration of cortisol in a time-depending manner. The addition of prednisolon stimulates the concentration of ACTH. CONCLUSIONS: The trophoblast cell shows a rhythmical course in the concentration of cortisol. For the first time a CRH-ACTH-cortisol feedback loop could be demonstrated in cultured trophoblast cells.

Adrenocorticotropic Hormone↗
PubMed2002

Complete bilateral crossed ureteral duplication observed during a radical hysterectomy with pelvic lymphadenectomy for ovarian cancer. A case report.

Complete bilateral ureteral duplication is a rare malformation. We report the case of a patient diagnosed with ovarian cancer, in which complete crossed bilateral ureteral duplication was observed during a planned radical hysterectomy with pelvic lymphadenectomy operation. If unilateral duplication is observed on one side, the existence of a similar malformation on the opposite side should always be suspected. Although bilateral ureteral duplication is an even rarer malformation than unilateral, it should be always considered as a differential diagnosis.

Acetabulum↗
PubMed2001

Fatal recurrent ureteroarterial fistulas after exenteration for cervical cancer.

BACKGROUND: Ureteroarterial fistula (UAF) is a rare occurrence. It can be difficult to diagnose with a high mortality. We report a case of a recurrent UAF. CASE: A 38-year-old women diagnosed with cervical cancer had undergone pelvic exenteration for severe radiation-induced necrosis with a vesicovaginal and rectovaginal fistula after primary radiation therapy. Hemorrhage into the urinary tract necessitated surgical intervention and vascular repair with a femoral-femoral bypass. Although these measures were effective, the patient died 6 months later following an acute hemorrhage into her conduit. Arteriogram revealed a second UAF. CONCLUSION: When urinary tract bleeding occurs in patients previously diagnosed with a gynecologic malignancy and treated with radiation therapy and extensive surgery with urinary diversion, UAF should be considered in the differential diagnoses.

Adult↗
PubMed2000

Immunohistochemical analysis of steroid receptors and glycodelin A (PP14) in isolated glandular epithelial cells of normal human endometrium.

Highly purified fractions of isolated endometrial cells can be useful for investigating endometrial function. After a first collagenase digestion, normal human endometrial stromal and epithelial cells were separated by filtration. Glands were purified further by two collagenase digestion steps, filtration, differential sedimentations, and Ficoll gradient centrifugation. Epithelial cells were polyhedral and grew as islands in a whorl-like wavy pattern around glandular fragments. High cell culture purity was confirmed with the positive immunohistochemical reaction against cytokeratin 7,8,18,19. Isolated human glands had a similar distribution pattern of estrogen receptor (ER) and progesterone receptor (PR) as observed in vivo, suggesting that glands have a functional hormone receptor system at the time of plating. Using a specific monoclonal antibody against glycodelin A (GdA), a characteristic cyclical expression was demonstrated during the menstrual cycle. The GdA reaction was weak in the proliferative phase, increasing significantly till the late secretory phase, suggesting a similar GdA concentration in vitro as observed in vivo glands. In conclusion, this method could be a model for studying endometrial glandular cells from different menstrual phases, endometrial cell interactions, implantation mechanisms, GdA regulation mechanisms, and pharmacological or other influences on ER and PR alteration.

Adult↗