Biomedical subjects

Klaus Friese

Publications and source records attributed to Klaus Friese.

At least 55 records · Page 3Linked to original sources

PubMed2004

Expression of the inhibin-alpha subunit in normal, hyperplastic and malignant endometrial tissue: an immunohistochemical analysis.

OBJECTIVES: Determination of the frequency and tissue distribution of inhibin-alpha (INH-alpha) in normal, hyperplastic and malignant endometrium. MATERIALS AND METHODS: Endometrial tissue was obtained from normal, hyperplastic (glandular-cystic hyperplasia), adenomatous hyperplasia (AH grade I to III) and endometroid adenocarcinoma and immunohistochemically characterized with INH-alpha antibody. RESULTS: INH-alpha was expressed in normal, hyperplastic and malignant endometrium. Highest expression was observed during secretory phase and glandular-cystic hyperplasia compared to all groups. A continuous decline was noted from AH grade I to III with a significance between AH I and II. DISCUSSION: A menstrual cycle associated expression of INH-alpha in glandular endometrial epithelium was observed. Since AH grade II and III can be considered as a precursor of endometrial cancer, INH-alpha could be a marker of cell transformation and an endometrial tumor-suppressor agent.

Biomarkers, Tumor↗

PubMed2004

Unexpected metastatic lobular carcinoma of the breast with intraabdominal spread and subsequent port-site metastasis after diagnostic laparoscopy for exclusion of ovarian cancer.

INTRODUCTION: Although lobular carcinomas metastasize primarily to lymph nodes, bone, lung and liver, they can also spread to the gastrointestinal tract, peritoneum and gynecologic organs. CASE REPORT: We report a case of intraperitoneal carcinomatosis of a lobular breast carcinoma that metastasized primarily to the peritoneum, with a subsequent abdominal wall invasion at the trocar site following laparoscopic surgery for the exclusion of an ovarian carcinoma. DISCUSSION: Port-site metastases (PSM) have occurred after laparoscopic surgery for endometrial, fallopian tube, ovarian, and cervical cancers. This is the first report of PSM of a lobular breast carcinoma primarily metastasized to the abdominal cavity. Every surgeon should be aware of the metastatic pattern of breast cancer, especially in relation to its histological subtypes. This case report emphasizes that PSM can occur in various kinds of gynecologic tumors, including breast cancer.

Aged↗

PubMed2004

N-glycans of human amniotic fluid transferrin stimulate progesterone production in human first trimester trophoblast cells in vitro.

AIMS: During pregnancy, the placenta produces a variety of steroid hormones and proteins. Several of these substances have been shown to exert immunomodulatory effects. Progesterone is thought to mediate some of these effects by regulating uterine responsiveness. The aim of this study was to clarify the effect of amniotic fluid transferrin and its N-glycans on the release of progesterone by first trimester trophoblast cells in vitro. METHODS: Cytotrophoblast cells were prepared from human first trimester placentae by trypsin-DNAse dispersion of villous tissue followed by a percoll gradient centrifugation and depletion of CD45 positive cells by magnetic cell sorting. Trophoblasts were incubated with varying concentrations (50-300 microg/ml) of transferrin from human amniotic fluid and serum as well as with N-glycans obtained from amniotic fluid transferrin. Culture supernatants were assayed for progesterone by enzyme-immunometric methods. RESULTS: The release of progesterone increased in amniotic fluid transferrin- and N-glycan-treated trophoblast cell cultures compared to untreated trophoblast cells. There was no stimulating effect of serum transferrin on the progesterone production of trophoblast cells. CONCLUSIONS: The results suggest that amnion-transferrin and especially its N-glycans modulate the endocrine function of trophoblasts in culture by up regulating progesterone secretion.

Amniotic Fluid↗

PubMed2003

Intra-mammary tumor location does not influence prognosis but influences the prevalence of axillary lymph-node metastases.

BACKGROUND: The number of axillary lymph-node metastases is not only a function of disease progression in primary breast cancer, but is also influenced by the intra-mammary location of the tumor. Nevertheless, the prognostic role of the tumor site is discussed controversially. The objective of this study was to analyze the impact of primary-tumor location on axillary lymph-node involvement, relapse, and mortality risk by univariate and multivariate analysis, in patients both with and without systemic and loco-regional treatment. METHOD: Retrospective analysis was conducted on 2,414 patients at the I. Frauenklinik, Ludwig-Maximilians University, Munich and Berlin-Charlottenburg, who underwent R(0) resection of the primary tumor and systematic axillary lymph-node dissection (at least five lymph nodes resected) for UICC I-III-stage breast cancer. Patients with unknown tumor site, multifocal tumor spread, central tumor location, or tumor location within 15 degrees of the border between outer and inner quadrants were excluded from the study. Median observation time was 6.7 years. RESULTS: The primary tumor site was within or between the medial quadrants of the breast in 33.6% of the patients ( n=810) and in the lateral hemisphere of the breast in 66.4% ( n=1,604). Tumor size, histopathological grading, and estrogen receptor status were balanced between patients with lateral and medial tumor location. Metastatic axillary lymph-node involvement was significantly associated with a lateral tumor location ( P<0.0001). The mean number of axillary lymph-node metastases was increased by 29% in cases with lateral tumor location (2.2 vs 1.7, P=0.003). In a multivariate logistic regression analysis allowing for tumor location, estrogen receptor status, grading and tumor size, tumor location was confirmed as a significant risk factor ( P=0.02) for axillary lymph-node involvement. Tumor location, however, did not correlate with either disease-free survival (DFS) or overall survival (OS), by univariate (DFS: P=0.41; OS: P=0.57) or by multivariate analysis (DFS: P=0.16; OS: P=0.98). CONCLUSION: We conclude that there is no sufficient evidence to support any independent prognostic significance of intra-mammary tumor location in early breast cancer. However, medial tumor location may lead to the underestimation of axillary lymph-node involvement.

Analysis of Variance↗

PubMed2003

Glycodelin and amniotic fluid transferrin as inhibitors of E-selectin-mediated cell adhesion.

Human amniotic fluid contains a variety of glycoproteins. Several of these substances have been shown to exert immunomodulatory effects. Glycodelin, previously known as placental protein 14, is one of these glycoproteins. It has a unique carbohydrate configuration, consistent with fucosylated LacdiNAc structures that are very unusual for mammals. Oligosaccharides with fucosylated LacdiNAc antennae have previously been shown to block selectin-mediated cell adhesion. Another glycoprotein, human transferrin, is also present in amniotic fluid in relatively high concentrations. This transferrin shows a different glycosylation compared with serum transferrin. Amniotic fluid transferrin carries sialylated Lewis X antigens. Glycodelin and transferrin were isolated from amniotic fluid and for comparison from serum of pregnant women by chromatographic methods. The purified proteins were used as ligands to block E-selectin-mediated HepG2 cell adhesion. Two types of binding assays with distinct receptor accommodations (immobilised E-selectin and activated HUVECs) were used to quantify inhibition efficiencies of the different proteins. We found that glycodelin is a strong inhibitor with a 10(3)-fold potency compared to the monovalent tetrasaccharide sialyl Lewis X whereas the potency of transferrin is rather low.

Adult↗

PubMed2003

The role of infection in preterm labour.

Although there are many maternal characteristics associated with preterm birth, the aetiology in most cases is poorly understood. Our research demonstrates that multiple risk factors, such as maternal age and especially infection, are associated with preterm birth. Bacterial vaginosis and intrauterine infection are now believed to be an important risk factor for preterm delivery.

Cost of Illness↗

PubMed2003

Nutrition and lifestyle factors on the risk of developing 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 therapy in postmenopause), inhibition of DNA-repair mechanisms (lack of vitamins), effecting estrogen metabolism (phytoestrogenes), or as promotors to enhance growth of tumours (body mass index). Some 'substances' may act as a carcinogenic itself, for example, 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.

Breast Neoplasms↗

PubMed2003

Perioperative screening for metastatic disease is not indicated in patients with primary breast cancer and no clinical signs of tumor spread.

BACKGROUND: Is a perioperative metastatic screening program indicated in patients presenting with primary operable breast cancer and no signs of distant metastases? PATIENTS AND METHODS: The impact of staging results (chest X-ray, bone scanning, liver ultrasound) for prognosis, treatment, quality of life and costs was retrospectively analyzed in 1076 patients with an operable breast cancer and no clinical signs of metastases. RESULTS: Staging examinations revealed 30 (2.8%) distant metastases, 130 (12.1%) suspect findings and excluded metastases in 916 (85.1%) patients. Further diagnostic procedures confirmed distant metastases in 7 (5.4%) and excluded them in 123 (94.6%) out of 130 patients with suspect findings. Distant metastases were detected more frequently with increasing pathological tumor size (pT < or = 2.0 cm: 1.6%, pT 2.1-5.0 cm: 3.0%, respectively pT > 5.0 cm: 15.1%; p < 0.001) and increasing number of involved axillary lymph nodes (pN0: 1.9%, pN1-3+: 1.8%, pN4-9+: 4.0%, pN > or = 10+: 18.7%; p < 0.001). Due to false positive findings 123 (11.4%) patients had to live for a significant period of time with the psychological distress of suspected metastatic disease. The abandonment of a perioperative screening in 1076 patients saves costs of at least Euros 259,367.68. CONCLUSIONS: In breast cancer patients without clinical signs of tumor spread perioperative screening for metastases is not warranted because of low frequency of metastases, false positive findings, missing therapeutic consequences and high costs.

Bone Neoplasms↗

PubMed2003

Patients with recurrent breast cancer: does the primary axillary lymph node status predict more aggressive tumor progression?

BACKGROUND: The extent of axillary lymph node involvement represents the foremost important prognostic parameter in primary breast cancer, and, thus, is one of the main determinants for subsequent systemic treatment. Nevertheless, the relevance of the initial axillary lymph node status on survival after disease recurrence is discussed controversially. Persisting prognostic impact after relapse would identify lymph node status as a marker for tumor biology, in contrast to a simply time-dependent phenomenon. METHOD: Retrospective analysis of 813 patients with locoregional or distant recurrence of primary breast cancer, who were primarily diagnosed with their disease at the I. Frauenklinik, Ludwig-Maximilians-University, Munich, and the University Hospital in Berlin-Charlottenburg, Germany, between 1963 and 2000. To be eligible, patients were required to have been treated for resectable breast cancer free of distant disease at the time of primary diagnosis, and must have undergone systematic axillary lymph node dissection. Patients with unknown tumor size or nodal status were excluded from the study. All data were gathered contemporaneously and compared with original patients files, as well as the local cancer registry, ensuring high quality of data. The median observation time was 60 (standard deviation 44) months. RESULTS: At time of primary diagnosis, 273 patients (33.6%) were node-negative, while axillary lymph node metastases were detected in 540 patients (66.4%). In univariate analysis tumor size, axillary lymph node status, histopathological grading, hormone receptor status, as well as peritumoral lymphangiosis and haemangiosis carcinomatosa were significantly correlated with survival after relapse (all, P < 0.0001). Kaplan-Meier analysis estimated the median survival time after relapse in node-negative patients to be 42 months (31-52 months, 95% CI), and 20 months in patients with 1-3 axillary lymph node metastases (16-24 months, 95% CI), compared to 13 months in patients with at least 4 involved axillary nodes (12-15 months, 95% CI). Multivariate logistic regression analysis, allowing for tumor size, axillary lymph node status, histopathological grading, presence of lymphangiosis carcinomatosa, relapse site and disease-free interval confirmed all parameters, except of histopathological grading (P = 0.14), as significant, independent risk factors for cancer associated death. Subgroup analyses, accounting for site of relapse and duration of disease-free interval, confirmed primary lymph node status as independent predictor for cancer-associated death after relapse. CONCLUSION: Lymph node involvement at primary diagnosis of breast cancer patients predicts an unfavorable outcome after first recurrence, independently of the site of relapse and disease-free interval. These observations support the hypothesis that primary lymph node involvement is not a merely time-dependent indicator for tumor progression, but indicates tumors with aggressive biological behavior.

Axilla↗

PubMed2003

Alterations in height, weight, and body mass index of newborns, children, and young adults in eastern Germany after German reunification.

OBJECTIVE: To examine the effects of the socioeconomic changes occurring during the German reunification on anthropometric data of newborns, children, and young adults. Study design Data were drawn from different perinatal surveys and cross-sectional studies that were conducted in the newly formed German states before and after reunification (1984-2000). RESULTS: The mean birth weight of newborns increased 151 g between 1984 and 1985 (n = 51,447) and 1997 (n = 83,052), whereas length increased by 0.2 cm. Among 7- to 10-year-old school children in the federal state of Saxonia between 1984 and 1985 (n = 2359) and 1999 and 2000 (n = 11,575), an increase of height by 1 to 2 cm and weight by 1.2 to 2.6 kg was observed. Average body mass index (BMI) increased by 0.7 to 0.9 kg/m(2). Among military recruits from Eastern Germany, an increase in weight and height was observed between 1985 and 1998. BMI values rose from 21.5 to 22.6 kg/m(2), and "centimeter weight" increased from 381 to 405 g/cm. CONCLUSION: The results show that the change in socioeconomic conditions was associated with alteration in anthropometric measures within a short period. This may have been caused by changes in the nutrient supply and composition as well as reduced physical activity.

Adult↗

PubMed2003

Cathepsin D expression in normal, hyperplastic and malignant endometrial tissue: an immunohistochemical analysis.

Cathepsin D (CathD), a lysosomal aspartyl protease secreted by normal and malignant cells, is considered to be involved in breakdown of the extracellular matrix. Aim of the present study was to determine the frequency and tissue distribution of CathD in normal, hyperplastic and malignant endometrium. Paraffin-fixed endometrial tissue was obtained from premenopausal women in the proliferative phase (n = 5), early secretory phase (n = 4) and late secretory phase (n = 4) as well as glandular-cystic hyperplasia (n = 5), endometrial polyps (n = 5), endometrial polyps from the use of tamoxifen (n = 5), adenomatous hyperplasia (AH) grade I (n = 5), grade II (n = 4), grade III (n = 5) and endometroid adenocarcinoma (n = 5). CathD expression was evaluated with the IRS score and ANOVA analysis was used for statistical evaluation. CathD was primarily localised in luminal and glandular epihelium with little staining in stromal cells. The expression of CathD was significantly higher during the late secretory phase than in the proliferative phase. Highest expression of CathD was observed in the late secretory phase and in glandular-cystic hyperplasia, whereas endometroid carcinoma showed no expression. A continuous increase in CathD expression was observed in AH, with a significant difference between AH grade I and III. In conclusion, CathD was found to be expressed in normal and hyperplastic endometrial tissue. CathD immunostaining in normal endometrial glands varied on the basis of the phase of the menstrual cycle, suggesting physiological functions of CathD in endometrial maturation and degradation. Adenocarcinomas did express significant lower amounts of CathD. Therefore, the prognostic value of this parameter remains uncertain. A continuous increase in CathD immunostaining was observed in AH. Since AH grade III can be considered as a precursor of endometrial cancer, CathD could be a possible parameter for assessing malignant transformation.

Cathepsin D↗

PubMed2003

Skin-sparing mastectomy with conservation of the nipple-areola complex and autologous reconstruction is an oncologically safe procedure.

OBJECTIVE: Is skin-sparing mastectomy (SSM) with conservation of the Nipple-Areola Complex (NAC) and immediate autologous reconstruction as safe in oncologic terms as SSM with resection of the NAC as modified radical mastectomy (MRM)? SUMMARY BACKGROUND DATA: The originally described technique of SSM included the removal of gland, NAC, and biopsy scar. However, the risk of tumor involvement of NAC in patients with breast cancer has been overestimated. PATIENTS AND METHODS: Between 1994 and 2000, 286 selected patients with an indication for MRM and tumor margins of greater than 2 cm from the nipple were presented with the alternative of a SSM. Regular follow-up data were evaluable of 112 patients with SSM and 134 patients with MRM. Immediate reconstruction was achieved by latissimus dorsi flap or TRAM flap. The mean follow-up time was 59 (18 to 92) months. RESULTS: Patients with SSM were significantly younger than those with MRM but were comparable regarding clinical data, tumor parameters, adjuvant treatment, and overall complications. After intraoperative frozen sections of the NAC-ground, the NAC could be conserved in 61 (54.5%) but was resected in 51 (45.5%) of the 112 patients with SSM. The aesthetic results after SSM were evaluated as excellent or good in 91.1% (102/112) patients and were significantly better after preservation of the NAC (P = 0.001). Six (5.4%) recurrences occurred in 112 patients with SSM compared with 11 (8.2%) cases after MRM. Only 1 recurrence in a conserved nipple was treated by wide excision of nipple with conservation of the areola. This patient is still free of disease after 52 months. CONCLUSION: In patients who are candidates for a mastectomy and tumors distant from the nipple, SSM with intraoperative frozen section of the NAC ground offers the opportunity of NAC conservation without increasing the risk of local recurrences.

Adult↗

PubMed2002

Stimulation of hCG and inhibition of hPL in isolated human trophoblast cells in vitro by glycodelin A.

The immunosuppressive protein glycodelin A (formerly named PP14) is produced by human decidua and secreted in the maternal circulation. Glycodelin A concentrations in serum have been used as indicators of endometrial function. The purpose of this study was to investigate the effect of glycodelin A on human chorionic gonadotropin (hCG) and human placental lactogen (hPL) release by freshly isolated cytotrophoblasts (in vitro). Cytotrophoblasts have been prepared from human term placenta by the three-step trypsin-DNase dispersion method of villous tissue followed by a percoll gradient centrifugation step. When placed in culture, the isolated mononuclear trophoblasts differentiated into syncytial counterparts within 12-72 h after plating. Trophoblasts were incubated with varying concentrations (60-300 microg/ml) of glycodelin A. Glycodelin A was isolated and purified by chromatographic methods from amnion fluid. Supernatants of the trophoblast cell cultures were assayed for hCG and hPL by immunological methods. The release of hCG is increased in glycodelin A-treated trophoblast cell cultures compared to untreated trophoblast cells. Glycodelin A inhibits the production of hPL in vitro. Differences in Glycodelin A stimulated cells and untreated controls are statistical significant. hCG and hPL are markers for the differentiation process of trophoblast cells to syncytial trophoblasts. The results imply that glycodelin A secreted by decidualised endometrium modulates endocrine function, as well as the differentiation of trophoblasts in culture.

Cells, Cultured↗

PubMed2002

Tumour Fas ligand:Fas ratio greater than 1 is an independent marker of relative resistance to tamoxifen therapy in hormone receptor positive breast cancer.

BACKGROUND: The objective of the present study was to examine the prognostic and predictive significance of the apoptosis-related marker Fas ligand (FasL):Fas ratio in breast cancer. METHODS: Tumour biopsies from 215 primary invasive breast cancer patients were examined for the expression of FasL and Fas mRNA transcripts by quantitative real-time RT-PCR. Their prognostic and predictive impact on patient survival was determined in univariate and multivariate survival analyses. RESULTS: Using a cutoff value of 1, a FasL:Fas ratio greater than 1 was found to have significant prognostic value for disease-free survival among the total population (median follow up 54 months). It was associated with a significantly decreased disease-free survival (P = 0.022) and with a tendency toward increased mortality (P = 0.14) in univariate analysis. Hormone receptor positive women exclusively treated with tamoxifen (n = 86) and with a FasL:Fas ratio greater than 1 had a significantly decreased disease-free survival (P = 0.008) and overall survival (P = 0.03) in univariate Kaplan-Meier analysis. Furthermore, tumour size and FasL:Fas ratio were of independent predictive significance in the multivariate model for disease-free and overall survival in that subgroup. Among postmenopausal patients (n = 148) both of those factors retained independent prognostic significance in the multivariate model for disease-free survival. In contrast, FasL:Fas ratio had no significant predictive value in patients exclusively treated with chemotherapy. CONCLUSION: The data presented indicate that FasL:Fas ratio may be useful not only as a prognostic factor but also as a predictive factor for projecting response to the antioestrogen tamoxifen. The results strongly support a correlation between FasL:Fas ratio greater than 1 and lack of efficacy of tamoxifen in hormone receptor positive patients.

Biomarkers, Tumor↗

PubMedDate not supplied

hCG in trophoblast tumour cells of the cell line Jeg3 and hCG isolated from amniotic fluid and serum of pregnant women carry oligosaccharides of the sialyl Lewis X and sialyl Lewis a type.

Human chorionic gonadotropin (hCG) is a placental hormone and marker for the differentiation process of cytotrophoblast cells to syncytial trophoblasts. It is secreted into the maternal circulation and urine. The glycoprotein hCG is also found in the serum and urine of patients with malignant trophoblastic diseases. For this study hCG was purified from pooled human mid-trimester amniotic fluid, serum and urine of pregnant women and from supernatants of trophoblast tumour cell cultures Jeg3 and BeWo by immunoadsorption chromatography with specific hCG-antibodies. The purity and identity of the isolated hCG were checked by Western blot analysis. Sialylated oligosaccharides of the Lewis type of isolated hCGs were analysed by dot blot and ELISA technique with monoclonal antibodies specific for sialyl Lewis x and sialyl Lewis a-carbohydrate epitopes. BeWo and Jeg 3 cells were additionally investigated by immunofluorescence. HCG isolated from the serum and amnion of pregnant women bear both sialyl Lewis x (sLex)-antigen and sialyl Lewis a (sLea)-antigen. The same result was observed for hCG isolated from the supernatants of the trophoblast tumour cell line Jeg3. On the contrary, hCG isolated from the supernatants of the trophoblast tumour cell line BeWo showed only a very weak expression of these antigens. HCG isolated from the urine of pregnant women was negative for both sialyl Lewis antigens. The results are discussed with respect to a possible relationship to selectin- mediated cell adhesion processes. This could play a role in the prevention of leukocyte adhesion on the foetal syncytiotrophoblast. Glycosylation of hCG in trophoblast tumour cells, on the other hand, could increase the metastatic activity of these cells.

Amniotic Fluid↗

PubMedDate not supplied

Stimulation of hCG protein and mRNA levels in trophoblast tumour cells Jeg3 and BeWo by glycodelin A.

The placental hormone human chorionic gonadotropin (hCG) represents a marker for the differentiation process of cytotrophoblast cells into syncytial trophoblasts and is also found in the serum and urine of patients with malignant trophoblastic diseases. During pregnancy, serum concentration curves of hCG and glycodelin A show a similar course. The main source of hCG is the trophoblast and trophoblast cells in vitro show an increased hCG release if treated with glycodelin A. In addition, hCG is a tumour marker for chorion carcinoma cells. We investigated the effect of native and recombinant glycodelin on the trophoblast tumour cells Jeg3 and BeWo and the role of expression plasmids of glycodelin A in the same cells. Our study shows that glycodelin A stimulates the secretion of hCG protein in Jeg3 trophoblast tumour cells in a time- and dose-dependent manner. Our results were confirmed on the mRNA level by real-time RT-PCR. The effect of glycodelin A on hCG mRNA regulation is time- and dose-dependent. We observed an increase of hCG mRNA copy numbers after glycodelin A treatment leading to a higher hCG protein production. Glycodelin A had no effect on BeWo trophoblast tumour cells, suggesting that production of hCG is not regulated by glycodelin A in these cells.

Chorionic Gonadotropin↗

PubMedDate not supplied

Induction of apoptosis in human breast cancer and trophoblast tumor cells by galectin-1.

Galectin-1 (gal-1), a member of the mammalian beta-galactoside-binding proteins, preferentially recognizes Galbeta1-4GlcNAc (LacNAc) sequences of oligosaccharides associated with several cell surface glycoconjugates. As demonstrated histochemically, gal-1 recognizes appropriate glycoepitopes on human breast cancer cells (MCF7) and on human chorionic carcinoma cells (BeWo). Gal-1 is expressed in many malignant and normal tissues. A high level of expression is found in lymphatic organs, which feature high rates of apoptosis. Furthermore, it is known that galectin-1 can initiate T cell apoptosis. In this study, we examined the apoptotic potential of gal-1 in vitro on MCF7 and BeWo cells. After growing both cell lines on chamber-slides for three days, apoptosis was induced by incubation with 30 microg gal-1 per ml serum-free medium for 6, 9 and 20 hours. To avoid false increased rates of apoptosis by deletion of FCS, all approaches were done with and without FCS. Apoptotic cells were detected by in situ nick translation. The rate of apoptosis was determined by counting 1500 cells per chamberslide. The normal rate of apoptosis ranged between 0.1% and 0.3%. The incubation of both cell lines with 30 microg/ml gal-1 in serum-free medium for 6 and 9 hours marginally raised the number of apoptotic cells. An increase of apoptosis was only shown by additional stimuli like hyperthermia, removal of CO2 and FCS for 20 hours. Impressing findings were manifested in an older passage of BeWo cells, in which only omission of FCS caused apoptotic rates for up to 25% after 6 hours. The presence of mycoplasma in this BeWo passage was shown by PCR. Our results demonstrated, that galectin-1 shows apoptotic potential in both the epithelial tumour cell lines examined only with additional stress stimuli.

Apoptosis↗

PubMedDate not supplied

Development of monoclonal and polyclonal antibodies and an ELISA for the determination of glycodelin in human serum, amniotic fluid and cystic fluid of benign and malignant ovarian tumors.

OBJECTIVE: The role of glycodelin in human reproduction and gynecological malignancies has been investigated in a large number of studies in recent years. Three dominant functions of this glycoprotein were identified. Glycodelin is immunosuppressive, a morphological marker of differentiation and a contraceptive. Glycodelin is a glycoprotein with a molecular weight of 28 kDa and a carbohydrate content of 17.5%. Unusual LacdiNAc structures were identified on glycodelin A, isolated from amniotic fluid. Because no kit for glycodelin quantification is commercially available, we developed all reagents and a functional ELISA. MATERIALS AND METHODS: Glycodelin A was purified from amniotic fluid by chromatographic methods. The purity of the isolated protein was checked with SDS-PAGE. Polyclonal antibodies against glycodelin were generated in rabbits. Monoclonal antibodies against glycodelin were generated from immunized BALB/c mice. Positive hybridomas were cloned and cultured. Monoclonal antibodies were isolated by immunoadsorption chromatography from culture supernatants. The glycodelin ELISA was developed in two formats, namely coating with polyclonal antibodies and the use of monoclonal antibodies. RESULTS: The factors of variance for the ELISA were 7% (intraassay variance) and 15% (inter-assay variance). The glycodelin ELISA was used to determine the glycodelin A concentration in sera of fertile women during the proliferative and secretory phases of the endometrium. The glycodelin A concentration was insignificantly elevated in the secretory phase compared to the proliferative phase. Significantly higher levels of glycodelin A were found in women using oral contraceptives compared to women who were not (p<0.001). This is probably due to progesterone, which stimulates glycodelin production. We also found significantly increased glycodelin concentrations in the fluids of malignant ovarian cysts compared to benign ovarian tumors (p<0.001). Furthermore, we tested the monoclonal and polyclonal antibodies successfully in Western blot analysis and immunoadsorption chromatography. CONCLUSION: We consider the described ELISA for the quantification of glycodelin as a useful tool for the determination of glycodelin in amniotic fluid, serum and cystic fluids. Its most promising application is expected in the diagnosis of ovarian cancer. The antibodies generated are applicable to multiple techniques.

Antibodies↗