Volker Briese

Complementary and alternative therapeutic approaches in patients with early breast cancer: a systematic review

SummaryComplementary and alternative medicine (CAM) is becoming increasingly popular, particularly among patients with breast cancer. We have done a systematic review of studies published between 1995 and February 2005, identified through a comprehensive search. CAM encompasses a wide range of treatment modalities, including dietary and vitamin supplements, mind-body approaches, acupuncture, and herbal medicines. The objectives of CAM treatments are diverse: reduction of therapy-associated toxicity, improvement of cancer-related symptoms, fostering of the immune system and even direct anticancer effects. Clinical trials have generated few or no data on the efficacy of CAM, whether regarding disease recurrence, survival, overall quality of life or safety. Some CAM methods may even have adverse effects or reduce the efficacy of conventional treatment. The primary justification for CAM is based on empirical evidence, case studies, and hypothetical physiological effects. We conclude that␣available data on CAM modalities in the treatment of early-stage breast cancer does not support their application.

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.

Maternal serum corticotropin-releasing hormone and ACTH levels as predictive markers of premature labor

Objective: To investigate whether corticotropin‐releasing hormone (CRH) and corticotropin (ACTH) plasma concentrations in women diagnosed with preterm labor are of potential clinical value in the assessment of the risk of preterm birth. Method: Plasma samples of 79 women diagnosed with preterm labor were used in this study. Samples were divided into three groups based on the week of gestation (24th–28th, 29th–32nd, 33rd–37th). CRH and ACTH values were determined by ELISA. Result: Mean maternal peripheral plasma values of CRH and ACTH were significantly higher (p < 0.001) in women who were initially diagnosed with preterm labor and finally delivered a preterm birth, compared to women with the same diagnosis but with term birth. Conclusion: CRH and ACTH serum levels in women diagnosed with preterm labor could be used as predictors for the timing of parturition.

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

BackgroundInhibins are multipotent dimeric glycoproteins, composed of an α-subunit and one of two possible β-subunits (βA or βB). Aims of this study were (a): the immunohistochemical characterisation of normal human endometrium for the inhibin-α 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 methodsSamples 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-α. 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.ResultsInhibin-α 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.DiscussionIn conclusion, we found a cyclical expression of inhibin-α 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.

Absolute quantification of human chorionic gonadotropin-β mRNA with TaqMan™ detection

We describe a reverse transcriptase-polymerase chain reaction (RT-PCR) for determination of human chorionic gonadotropin-β (HCGβ) mRNA copies using the TaqMan™ system. To evaluate our quantitative assay, we analyzed HCGβ transcripts of all protein coding genes (HCGβ 5, 3, 8, and 7) in human RNA panels of different normal tissues and in glycodelin-A-stimulated trophoblast cell cultures. Absolute quantification using HCGβ TaqMan probe was found to be highly reproducible. Our study of RNA panels confirms recently published results that expression of HCGβ transcripts is a common feature of a great variety of different normal tissues. High levels of HCGβ mRNAs (>1.000 molecules per 200 ng RNA) were detected in placenta, uterus, and testis. An increase of HCGβ mRNA expression (1.7-fold) was detected at 150 µg/mL glycodelin-A treatment in trophoblast cell culture. Time-dependence study showed that the increase in HCGβ mRNA level was evident at 60 min after glycodelin-A treatment. In summary, we have developed a highly sensitive one-tube, one-enzyme quantitative RT-PCR system that is time-saving and avoids postamplification procedures.

Obesity and pregnancy--a risk profile.

AIM We aimed to illustrate the relationship between maternal obesity during pregnancy and maternal and fetal outcomes. We examined the influence of maternal BMI at the beginning of pregnancy on risks of pregnancy and birth, and on the somatic classification of the neonates. MATERIAL AND METHODS In our retrospective cohort study we included 499,267 singleton pregnancies taken from the German perinatal statistics of 1998-2000. 51,506 obese pregnant women (BMI >or= 30) were compared to 320,148 pregnant women of normal weight (BMI 18.50-24.99). We divided obesity into 3 BMI-categories: BMI = 30.00-34.99, BMI = 35.00-39.99, and BMI >or= 40.00. We defined small-for-gestational-age (SGA), appropriate-for-gestational-age (AGA), and large-for-gestational-age (LGA) status by birth weight percentiles. RESULTS 10.3 % of all pregnant women had a BMI >or= 30.00 and 0.8 % had a BMI >or= 40.00. The frequency of hypertension increased with the extent of obesity: 7.1 % (BMI = 30.00-34.99), 12.5 % (BMI = 35.00-39.99) and 18.3 % (BMI >or= 40.00) compared to 1.2 % (BMI 18.50-24.99). Cephalopelvic disproportion was found in 6.8 % (BMI >or= 40.00) compared to 2.8 % (BMI 18.50-24.99). Fetal macrosomia occurred in 24.8 % (BMI >or= 40.00) compared to 7.9 % in the control group. Rates of pre-eclampsia, gestational diabetes, and fetal structural anomalies also increased with maternal BMI. Women with different BMIs differed in parity but not in age. CONCLUSIONS Obesity during pregnancy is associated with a range of maternal and fetal adverse outcomes. Pregnancy in obese women therefore calls for close monitoring and careful planning of delivery. Pre-conceptional weight reduction should be considered.

SERUM PLACENTAL PROTEIN 14 (PP14) LEVELS IN PATIENTS WITH THREATENED ABORTION

Abstract. Serum placenta protein 14 (PP 14) were significantly lower serum levels in patients with threatened abortion at 10 to 20 weeks controls than in normal (normal group, n = 133, median: 52,0 µg/l [16 – 83, SD: 17,9], risk group: n = 20, median: 34,0 µg/l [13 – 63, SD: 13,7]).

Influence of phytoestrogens on the proliferation and expression of adhesion receptors in human mammary epithelial cells in vitro.

Tumor metastasis is associated with integrin-mediated adhesion and hyaluronan receptor expression. Accumulating evidence suggests that phytoestrogens, which are naturally occurring, plant-derived phytochemicals, could inhibit tumorigenesis during the development of breast cancer. Less is known, however, about the regulation of adhesion receptors by phytoestrogens and, particularly, their potency to influence proliferation of primary human breast cells in comparison with the steroid hormone 17&bgr;-estradiol. Throughout the proliferation experiments, we used primary human mammary epithelial cells from normal tissue that was derived from plastic surgery. For receptor expression (&bgr;1, &agr;2, &agr;3, CD44), we used the cell line MCF-7. Both investigations were carried out by flow cytometry. The phenotype of primary human mammary epithelial cells was microscopically characterized by analyzing the distribution of ZO-1, cytokeratin and the estrogen receptors &agr; and &bgr;. The integrins and the hyaluronan receptor were significantly up-regulated with 17&bgr;-estradiol in human MCF-7 cells. In contrast, genistein and daidzein did not affect the expression at a concentration of 100 μmol/l. In all proliferation experiments with a significant stimulation of the primary human mammary epithelial cell growth due to 17&bgr;-estradiol, in general, genistein and daidzein did not influence S-phase and G2/M-phase cells. Additionally, the stimulative effect of 17&bgr;-estradiol could be inhibited. As the phytoestrogens do not up-regulate adhesion receptors in human breast cells and, regarding proliferation, are able to abolish the stimulatory effect of 17&bgr;-estradiol, we suggest that phytoestrogens could have beneficial effects for the prevention or inhibition of carcinogenesis in hormone-dependent malignancies.

Effects of phytoestrogens genistein and daidzein on progesterone and estrogen (estradiol) production of human term trophoblast cells in vitro

Objective. Phytoestrogens are a diverse group of nonsteroidal plant compounds that occur naturally in many plants. Because they possess a ring system similar to estrogens they are able to bind on estrogen receptors alpha and beta in humans. The effects of the phytoestrogens genistein and daidzein on the production of progesterone and estrogen in isolated human term trophoblast cells in vitro were tested in this study. Material and methods. Cytotrophoblast cells were isolated from human term placentas. Phytoestrogens genistein and daidzein were incubated in different concentrations with trophoblast cells. Untreated cells were used as controls. After 24 h aliquots were removed and tested for progesterone and estrogen production. Results. The production of the steroid hormones progesterone and estrogen are influenced by phytoestrogens genistein and daidzein in human term trophoblast cells. A strong inhibition effect of both phytoestrogens tested in the production of progesterone was demonstrated. In addition, a significant stimulating effect on estrogen production by genistein and daidzein was observed. Conclusion. Results obtained with this study show that phytoestrogens (genistein and daidzein) sufficiently reduce progesterone production in human term trophoblast cells. Because blockade of progesterone is a possible mechanism involved in initiation of labor, we may speculate that high doses of phytoestrogens at the feto-maternal interphase could play a negative role in maintenance of pregnancy. Stimulation of estrogen production by genistein and daidzein in trophoblast cells is probably due to estrogen receptor blocking effects of both phytoestrogens. Trophoblast cells seem to compensate blocking of its estrogen receptors by higher estrogen production.

Stimulation of hCG protein and mRNA in first trimester villous cytotrophoblast cells in vitro by glycodelin A

Abstract Aim: Human chorionic gonadotropin (hCG) is produced by fetal trophoblast cells and secreted into maternal circulation mainly in the first trimester of pregnancy. Another glycoprotein, glycodelin A, is one of the main products of the maternal decidua during this period. The purpose of this study was to investigate the effect of glycodelin A on hCG release by isolated cytotrophoblast cells in vitro. Methods: Cytotrophoblast cells were prepared from human first trimester placenta and incubated with varying concentrations of glycodelin A. Supernatants were assayed for hCG protein concentrations, and quantification of beta hCG mRNA was carried out by RT-PCR. Expression of hCG was analysed in stimulated trophoblast cells and in unstimulated controls by immunocytochemistry. Results: Glycodelin A induces a dose-dependent increase of hCG production. An increase of hCG expression was measured at 100 and 200 μg/mL glycodelin-A treatment in trophoblast cell culture by TaqMan assay on mRNA level. We found a moderate staining of hCG in control trophoblast cells, whereas a strong hCG staining was seen in glycodelin A-treated trophoblast cells. Conclusions: HCG is a marker for the differentiation process of trophoblast cells. Our results suggest that glycodelin A secreted by the decidualized endometrium is involved in the regulation of hormones produced by the trophoblast.