Relation of myeloperoxidase promoter polymorphism and long-term hormone replacement therapy to oxidized low-density lipoprotein autoantibodies in postmenopausal women.

OBJECTIVE: The myeloperoxidase enzyme (MPO) is a potent precursor of low-density lipoprotein (LDL) oxidation in atherosclerotic lesions. The MPO gene has a promoter polymorphism, 463G/A, which leads to high (GG) and low-expression (AG, AA) genotypes. Hormone replacement therapy (HRT) is known to affect MPO activity and LDL oxidation. The purpose of this study was to test whether the effect of HRT on the levels of oxLDL-ab varies according to MPO genotype. MATERIAL AND METHODS: Eighty-seven postmenopausal women aged 45-71 years were divided into three groups based on the use of HRT. The HRT-EVP group (n = 25) used sequential estradiol valerate (EV) plus progestin, the HRT-EV group (n = 32) used EV alone, and the control group (n = 30) no HRT. MPO genotypes were determined by polymerase chain reaction (PCR) and oxLDL-ab by ELISA. RESULTS: We found a significant HRT group by MPO genotype interaction (p = 0.021) in plasma oxLDL-ab levels. In subjects with the GG genotype, the oxLDL-ab titer increased in the order of 2.13 in controls, 2.53 in the EV group and 3.21 in the EVP group (ANOVA for trend p = 0.006). CONCLUSIONS: The effects of HRT on LDL oxidation can vary according to MPO genotype and the concurrent progestin therapy with EV may counteract the more neutral effect of EV on LDL oxidation in subjects with the MPO high-expression genotype.

The relation of oxidized LDL autoantibodies and long-term hormone replacement therapy to ultrasonographically assessed atherosclerotic plaque quantity and severity in postmenopausal women.

BACKGROUND: In epidemiologic studies, the incidence of atherosclerosis rises soon after menopause in women, and hormone replacement therapy (HRT) has proved to be useful in preventing onset of clinical manifestations of the disease. However, it is not known how HRT affects sonographically determined atherosclerotic severity (AS) and number of atherosclerotic plaques (NAP) in large arteries. Furthermore, it is not clear how HRT affects oxidation of low density lipoproteins (LDL), which obviously has an important role in the pathogenesis of atherosclerosis. OBJECTIVES: The purpose of the study was to determine whether HRT has a beneficial effect on sonographically determined AS and NAP in large arteries of 101 postmenopausal women compared to 40 controls without HRT. We also studied the interaction of HRT and antibodies against oxidized LDL on AS and NAP progression. RESULTS: Estradiol valerate alone, combined estradiol valerate-levonorgestrel and combined estradiol valerate-medroxyprogesterone acetate therapy are each associated with lower NAP and AS as compared to controls without HRT. In a multiple regression model explaining NAP in the whole study population, the strongest predictors were HRT (P=0.0006) and copper-oxidized LDL cholesterol autoantibodies (P=0.0491). DISCUSSION: Our findings indicate that postmenopausal HRT is associated with a lower total number of atherosclerotic plaques and less severe atherosclerotic lesions, as compared to controls without HRT, and that this outcome may be associated with the effect of HRT on LDL cholesterol oxidation.

Magnetic resonance imaging based volume estimation of ovarian tumours: use of a segmentation and 3D reformation software.

The application of a new segmentation software, Anatomatic in the evaluation of volumetric measurements of ovarian tumours and the new Medimag three-dimensional (3D) software in the evaluation of 3D image representation of ovarian tumours with 1.5 T magnetic resonance imaging (MRI) is described. Our goal was to compare MRI based volumetry with operative findings at laparotomy for six consecutive patients with suspected ovarian tumours. Volumetric analysis and three dimensional image reconstructions of the tumours were obtained. At laparotomy, the tumour sizes were measured in situ, and the volumes were calculated. Using Anatomatic, reproducible tumour volumes were achieved with ease and within a reasonably fast time in patients with ovarian tumours without ascites. Medimag helped achieve realistic 3D representations of the tumours. For the four solitary tumours segmentation based volumetry and laparotomy findings agreed in three cases. In one patient with an oval shaped tumour, the segmented volume was double as compared to that estimated at laparotomy. Of the two patients with multiple tumours, both patients had significant ascites, and volumetry misinterpreted the fluid as tumour cyst fluid and markedly overestimated the tumour size. In conclusion, the MRI based segmentation volumetry and 3D image reconstructions are rapid, and reproducible methods of measuring ovarian tumours in patients without significant ascites.

Is potential to produce inhibins related to prognosis in ovarian granulosa cell tumors?

OBJECTIVE: The prognosis of ovarian granulosa cell tumors is difficult to predict by conventional histological methods. STUDY DESIGN: The tumors of 30 patients operated on for a granulosa cell tumor were retrospectively studied for the expression of the inhibin alpha-subunit. The immunohistochemical staining results were correlated to the FIGO stage of the tumor and the prognosis of the patients. RESULTS: Twenty-six (87%) of the ovarian granulosa cell tumors stained positively for the inhibin alpha-subunit. All FIGO stage I and II tumors expressed inhibin alpha-subunit. All FIGO stage I and II tumors expressed inhibin, whereas the majority of stages III and IV did not (p=0.001, chi2-test). The survival of the patients with inhibin immunopositive tumors was significantly longer (median 183 months, SE 41, p=0.000, log rank test) than that of patients without inhibin expression (median 2.5 months, SE 5.25). CONCLUSIONS: Patients with ovarian granulosa cell tumors with a potential to produce the inhibin alpha-subunit may have a more favorable prognosis than those with inhibin immunonegative tumors.

Magnetic resonance imaging compared to ultrasonography in fetal weight and volume estimation in diabetic and normal pregnancy.

OBJECTIVE: To estimate fetal volume and weight in diabetic and normal pregnancy using high-resolution magnetic resonance imaging. METHODS: T1-weighted magnetic resonance imaging was combined with semiautomatic segmentation technique. The accuracy of fetal volume estimations thus obtained was compared with conventional ultrasound-based weight estimations in ten pregnant women with insulin-dependent diabetes mellitus and ten women with normal pregnancy. Examinations were made within 48 hours before delivery. RESULTS: Ultrasound-based estimations of fetal weight showed a correlation rate of r=0.77 with the actual birth weights in the whole material, while volume determinations based on magnetic resonance imaging showed a significantly better correlation rate of r=0.95. Diabetic women did not differ from the normal pregnancy group with regard to birth weight or the accuracy of weight estimations. CONCLUSIONS: High-resolution magnetic resonance imaging combined with semiautomatic segmentation software was found to be accurate in determining fetal volume and, consequently, better than conventional ultrasound-based techniques in estimating fetal weight. The use of magnetic resonance imaging in fetal weight estimation may be recommended for clinical situations where an accurate weight estimate is considered essential.

Inhibin A, B and pro-alphaC in serum and peritoneal fluid in postmenopausal patients with ovarian tumors.

OBJECTIVE: To compare serum and peritoneal fluid concentrations of inhibin A, B, and pro-alphaC in women with ovarian tumors. METHODS: Serum and peritoneal fluid samples were taken from 41 postmenopausal women operated on for an ovarian tumor. Twenty-one patients with endometrial cancer formed a control group. Serum and peritoneal fluid inhibin A, B, and pro-alphaC concentrations, and serum FSH and tumor marker CA 125 (study group only) concentrations were analyzed. RESULTS: Inhibin A was found in low concentrations (median 4.1pg/ml, range <2-29pg/ml) in serum in most postmenopausal patients with epithelial ovarian carcinoma, whereas inhibin B was not measurable. Inhibin pro-alphaC circulated in high concentrations (median 125pg/ml, range 37->1000pg/ml). All inhibins were found in clearly greater concentrations in the peritoneal fluid than in serum. International Federation of Gynecology and Obstetrics (FIGO) stage III-IV and poor differentiation grade were associated with significantly lower concentrations of inhibin A and pro-alphaC in the peritoneal fluid compared with stages I-II or low grade. This correlation was not found in the serum concentrations of inhibin A or pro-alphaC. In the control group, no dimeric inhibins were found in serum, and pro-alphaC circulated in median concentrations of 47pg/ml (range 12-174pg/ml). CONCLUSIONS: Postmenopausal patients with epithelial ovarian tumors had low concentrations of inhibin A and relatively high concentrations of inhibin pro-alphaC in serum. The peritoneal fluid concentrations of all inhibins far exceeded those in the serum. Relatively low concentrations of inhibin A and pro-alphaC in the peritoneal fluid of patients with ovarian cancer seem to be associated with high stage and grade and, to a lesser degree, with positive peritoneal cytology.

Regulation of UT-OC-3 ovarian carcinoma cells by cytokines: inhibitory effects on cell proliferation and activation of transcription factors AP-1 and NF-kappaB.

The present study was designed to investigate the growth regulatory effects of cytokines in UT-OC-3 ovarian cystadenocarcinoma cells in vitro. The effects of interleukin-6 (IL-6), interferons alpha (IFN-alpha) and gamma (IFN-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor alpha (TNF-alpha), and transforming growth factor beta1 (TGF-beta1) were investigated by (125)I-deoxyuridine ((125)IUdR) incorporation assay. In order to understand better the molecular mechanisms of the observed effects, the activation of DNA-binding proteins was studied by electrophoretic mobility shift assay. In addition, cellular DNA was tested by fragmentation analysis to determine if the most growth inhibitory cytokines are able to induce programmed cell death (apoptosis). After 48h in culture, TGF-beta1, TNF-alpha, IFN-alpha and IL-6 showed a clear inhibitory effect on (125)IUdR incorporation (P<0.005), and IFN-gamma and GM-CSF caused even more significant inhibition (P<0.001). IFN-alpha and IFN-gamma were both growth inhibitory after 72h in culture (P<0.005). Similarly, GM-CSF induced a slight inhibition (P<0.05), whereas TGF-beta1 and TNF-alpha almost blocked DNA synthesis (P<0.001) after 72h. IL-6 had no statistically significant effect on cell proliferation after 72h. Transcription factors AP-1 and NF-kappaB were both constitutively expressed in UT-OC-3 cells. The binding activity of AP-1 was found to be stimulated by the growth inhibitory cytokines, TGF-beta1 and TNF-alpha, and the binding of NF-kappaB was stimulated by TNF-alpha. Apoptosis does not seem to be induced by any of these cytokines in the UT-OC-3 ovarian cancer cell model.

Ovarian cancer and gonadotropins in vitro: new evidence in favor of independence.

BACKGROUND: The aim of this study was to investigate whether newly established epithelial ovarian carcinoma cell lines secrete inhibins, and whether their proliferative and secretory activity can be regulated by gonadotropins. MATERIALS AND METHODS: Three recently characterized human epithelial ovarian carcinoma cell lines were exposed to human choriongonadotropin hCG and follicle stimulating hormone FSH. Cell proliferation was determined by counting. Secretory activity of the cell lines was studied by analyzing inhibin A, inhibin B, inhibin pro-aC, estradiol, progesterone, testosterone, and CA 125 concentrations from the medium. The expression of gonadotropin receptors was studied by RT-PCR. RESULTS: None of the cell lines were found to secrete any of the inhibins, progesterone or testosterone. Only UT-OC-4 cells secreted low levels of estradiol. Gonadotropin receptors were not expressed by any of the cell lines, and accordingly neither FSH nor hCG stimulated the growth of these cells. However, hCG had some dose dependent growth inhibitory effect on UT-OC-3. Passage 42 cells of UT-OC-3 secreted significantly more CA 125 than passages 8 cells (P = 0.000). CONCLUSIONS: The present results suggest that the carcinomatous epithelial cells of the ovary do not secrete inhibins. The serum inhibin levels previously detected in patients with epithelial ovarian carcinoma may therefore reflect an ovarian stromal response to carcinoma. The findings are also in favor of an independence of ovarian cancer of gonadotropins.

Ovarian testosterone secretion during perimenopause.

OBJECTIVE: The aim of the study was to investigate ovarian testosterone secretion during the last few years of reproductive life and after menopause. MATERIALS AND METHODS: Ovarian and peripheral venous levels of total testosterone were analyzed in 52 women aged 42-69 years (mean 51) undergoing hysterectomy with adnexal removal for benign indications at the Department of Obstetrics and Gynecology at Tampere University Hospital, Finland. The study population was divided into pre- (n = 19), peri- (n = 18) and postmenopausal (n = 15) women in addition to the classical division according to menstrual cycle. Corresponding serum estradiol, progesterone and gonadotropin levels were measured, and the degree of ovarian stromal hyperplasia was analyzed. RESULTS: The levels of all steroid hormones were higher in the ovarian vein than in the periphery. A significant positive correlation was found between age and ovarian vein testosterone levels (r = 0.3, P = 0.01). In premenopausal women, it was 1.5 nmol/l (median; range 0.78-6.0), in perimenopausal women 2.2 nmol/l (range 0.9-13.6), and 2.5 nmol/l (range 0.6-26.6) in postmenopause, respectively. Peripheral testosterone level did not increase with age. Ovarian stromal hyperplasia was significantly associated with increased testosterone secretion (P = 0.009). CONCLUSION: The ovary seems to increase the secretion of testosterone into circulation during the menopausal transition period, as the highest levels were measured in postmenopausal women. High testosterone levels in the ovarian vein, however, were not reflected in the peripheral venous blood.

Cytokine production profiles in the peritoneal fluids of patients with malignant or benign gynecologic tumors.

BACKGROUND: Cytokines play a key role in the regulation of cells of the immune system and also have been implicated in the pathogenesis of malignant diseases. Some cytokines have been shown to have potential in the diagnosis of cancer. METHODS: A total of 111 patients with ovarian, cervical, or endometrial carcinomas or benign ovarian or uterine tumors were enrolled on the study, and the levels of interleukin (IL)-2, IL-4, IL-6, IL-10, interferon (IFN)-gamma, granulocyte-colony stimulating factor (G-CSF), granulocyte-macrophage-colony stimulating factor (GM-CSF), macrophage-colony stimulating factor (M-CSF), and tumor necrosis factor (TNF)-alpha were measured by cytokine specific, enzyme-linked immunoadsorbent assays. In addition, ratios of IL-2, IL-4, and IFN-gamma production were studied to characterize the type of T-cell response that occurred in the peritoneal cavities of the patients. RESULTS: High levels of M-CSF (mean for all patients, 26,050 pg/mL) and G-CSF (mean for all patients, 20,267 pg/mL) were observed in virtually all patients, but no significant differences between the study groups were observed. Similarly, no differences in the levels of IL-2, IL-4, IL-10, IFN-gamma, GM-CSF, or TNF-alpha were found. However, IL-6 levels were significantly higher in patients with ovarian carcinoma (mean +/- standard error of the mean [SEM]: 5572 +/- 1266) or benign tumors (mean +/- SEM: 4474 +/- 2008) than in those with cervical (mean +/- SEM: 1222 +/- 546) or endometrial carcinoma (mean +/- SEM: 1977 +/- 616). A predominantly Th1 type cytokine profile, irrespective of the diagnosis, was observed in patients with gynecologic tumors. CONCLUSIONS: With the exception of IL-6, the cytokine synthesis profiles in the peritoneal fluids of patients with benign and malignant gynecologic tumors were found to be similar. These results suggest that cytokine production in these patients is a result of nonspecific inflammation rather than a specific response against the tumor cells, and that skewing of cytokine synthesis toward either the Th1 or the Th2 phenotype is not the underlying mechanism resulting in the malignant process in women with gynecologic tumors.

Activin B: detection by an immunoenzymometric assay in human serum during ovarian stimulation and late pregnancy.

A recently developed immunoenzymometric assay for activin B has been characterized further by measurement during ovarian stimulation and pregnancy. The assay is based on a monoclonal anti-peptide antibody, anti-betaB(101-115). In addition to quantitative analyses, the antibody has been used for immunohistochemical localization of the activin betaB-subunit in human term placenta. Serum samples obtained from patients suffering from tubal factor infertility who were admitted for in-vitro fertilization (IVF) treatment protocols or from patients with proven fertility who were admitted for laparoscopic tubal ligation were collected. The aim was to correlate serum activin B concentrations with other parameters during IVF and with phases of the menstrual cycle. Serum samples obtained from healthy pregnant volunteers were studied to correlate activin B concentrations with clinical parameters. During the IVF treatment protocols, activin B was detectable in all patients studied, and a significant negative correlation was observed between serum activin B and oestradiol concentrations. On the other hand, no significant difference was observed in activin B concentrations when serum samples obtained from patients at different phases of the menstrual cycle were compared, and low concentrations of activin B were observed in the samples obtained from these patients. During pregnancy, a positive correlation was observed between serum activin B concentrations and gestational age. In immunohistochemical analyses of human placental tissue obtained from healthy parturients, the activin betaB-subunit was present in trophoblast, amniotic epithelial and Hofbauer cells. The results suggest a potential clinical application in female reproductive medicine for serum activin B measurements.

Inhibin A and B in peri- and postmenopause.

OBJECTIVE: The aim of the study was to investigate the origin of inhibin A and B during the last years of the reproductive age and after menopause by measuring their levels in the ovarian and peripheral venous blood. METHODS: The study population consisted of 43 women, aged 42-69 years (mean 50), who underwent hysterectomy with ovarian removal for a benign disease. A total of 24 of them were in follicular phase, 11 in luteal phase, and eight were postmenopausal. Peripheral and ovarian venous blood was collected for measurement of inhibin A and B. In addition, sex steroid hormone and gonadotropin levels were measured. RESULTS: Ovarian venous inhibin B correlated significantly with ovarian estradiol secretion (r = 0.5, P = 0.001). The levels of inhibin B were significantly higher in the ovarian vein than in the peripheral vein (P = 0.006). The highest inhibin B concentrations were detected in the mid-proliferative (mid-follicular) phase (median 31.6 pg/ml range 25.9-47.9). In postmenopausal women, inhibin B was not detectable. No correlation between FSH and ovarian inhibin B was found. Inhibin A rose rapidly in late proliferative (late follicular) phase (median 28.5, range < 2-51.8) and dominated in the circulation throughout the luteal phase (median 20.9, range 8.8-60). For inhibin A, no concentration gradient existed between the ovarian and peripheral vein. Unlike inhibin B, inhibin A was detectable in ovarian and peripheral blood in postmenopausal women. A significant negative correlation between ovarian and peripheral inhibin A and FSH was found (r = -0.386, P = 0.015; r = -0.345, P = 0.034, respectively). CONCLUSION: Inhibin B correlates with ovarian estradiol secretion and seems to reflect follicular function. Inhibin A dominates in circulation during the luteal phase but is detectable at low concentrations both in follicular phase and even in postmenopause. Our findings suggest that inhibin A may play a role in FSH suppression in the female reproduction. In addition to the ovary, there may be extragondal source(s) of inhibin A.

Inhibitory effects of cytokines on ovarian and endometrial carcinoma cells in vitro with special reference to induction of specific transcriptional regulators.

In the present study, we have investigated the effects of interferons-alpha (IFN-alpha) and -gamma (IFN-gamma), interleukin-10 (IL-10) and -13 (IL-13), transforming growth factor-beta1 (TGF-beta1), granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha) on cell proliferation and induction of transcription factors AP-1 and NF-kappaB in UM-EC-3 human endometrial adenocarcinoma cells and UT-OC-5 ovarian carcinoma cells in vitro. In addition, cellular DNA was extracted to study if any of these factors is able to induce apoptosis. In UM-EC-3 cell line DNA synthesis was inhibited by GM-CSF, IL-10, IL-13, TGF-beta1, IFN-alpha, and IFN-gamma after 48 and 72 h in culture, whereas TNF-alpha had no significant effect on cell proliferation in any of the experiments. The inhibition of DNA synthesis was similarly observed in UT-OC-5 ovarian carcinoma cells by IL-10, TNF-alpha, and IFN-gamma after 48 and 72 h, whereas IFN-alpha had no statistically significant effect. An inhibitory effect of GM-CSF was observed only after 48 h and TGF-beta after 72 h in culture, respectively. Transcription factors AP-1 and NF-kappaB were both constitutively active in UM-EC-3 and UT-OC-5 cells. The binding activity of AP-1 was found to be stimulated by all growth-inhibitory cytokines studied in both cell lines, whereas the specific binding activity of NF-kappaB was affected moderately only by TNF-alpha in UT-OC-5 ovarian carcinoma cells. No signs of DNA fragmentation typical of apoptosis were observed in any of these studies.

Prognostic significance of p53 expression in ovarian granulosa cell tumors.

OBJECTIVE: The purpose of the study was to investigate the significance of p53 expression for the prognosis in patients with ovarian granulosa cell tumors (GCT). METHODS: The records of 30 patients operated on for GCT at Tampere University Hospital, Finland, were reviewed. The mean age at the time of the diagnosis was 55 years. Twenty-one of the tumors were of FIGO stage I, three were of stage II, three were of stage III, and three were of stage IV. Paraffin-embedded tumor specimens were analyzed by immunohistochemistry for expression of mutated p53 protein and by flow cytometry. RESULTS: Eleven tumors were positive for p53 and 19 were negative. The median crude survival of p53-negative patients was 10 times that of p53-positive ones (210 months vs 21 months, P = 0.037, log-rank test). The association between p53 immunoreactivity and stage was statistically significant (P = 0.026 Pearson chi2 test), while there was no association between p53 expression and DNA ploidy or S-phase fraction. CONCLUSION: Although the results should be considered as preliminary, expression of mutated p53 in ovarian granulosa cell tumors seems to be associated with unfavorable prognosis.

Impaired ovarian function and risk factors for atherosclerosis in premenopausal women.

OBJECTIVES: The aim of the study was to obtain information on the possible relationship between impaired ovarian function and risk factors for cardiovascular disease. MATERIAL AND METHODS: Serum lipid levels, plasma fibrinolytic potential and histological and biochemical changes in the intima of the uterine artery were investigated in premenopausal women with irregular menstrual cycles, and the results were compared with those from regularly menstruating women. In addition, the same parameters were studied in postmenopausal women on hormone replacement therapy (HRT) and in postmenopausal women who had never used HRT. In total 64 patients undergoing hysterectomy for benign reasons were included the study. RESULTS: Plasma fibrinogen concentration was significantly higher in irregularly menstruating women as compared with women with regular cycles. In women with irregular cycles thickened or sclerotic arterial intima was a significantly more common finding as compared with regularly menstruating women. A significant positive correlation was observed between plasma fibrinogen concentration and intimal esterified cholesterol content in women with thickened or sclerotic uterine artery. CONCLUSIONS: These data suggest an important role for normal ovarian function in the prevention of atherosclerosis.

Regulation of proliferation of UM-SCV-1A and UM-SCV-6 vulvar carcinoma cells by cytokines.

The biology and pathogenesis of vulvar carcinoma are poorly understood at present. In order to understand this disease better, we have used recently developed squamous cell carcinoma lines of the vulva as models. Two cell lines originating from two individuals (UM-SCV-1A and UM-SCV-6) were cultured in vitro in 10% fetal calf serum. The effects of interleukins 10 and 13, interferons alpha and gamma, granulocyte/macrophage-growth-stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF alpha), and transforming growth factor beta (TGFbeta) on the proliferation of the cells was investigated by using radioactively labelled uridine as tracer. In addition, an investigation on the molecular structure of extracted cellular DNA was carried out to investigate whether programmed cell death (apoptosis) would be inducible by any of the factors. In UM-SCV-1A cells, interleukin-10 (IL-10) and interleukin-13 (IL-13) caused an approximately 12-fold decrease in DNA synthesis in cells cultured for 72 h (P<0.001), while GM-CSF had no significant effect. TGFbeta showed a significant inhibitory effect on deoxyuridine incorporation (P<0.001), which was 2.0- and 4.2-fold at 48 h and 72 h, respectively. TFG alpha showed a 1.2-fold inhibitory effect on DNA synthesis at 48 h (P<0.01) and a 1.5-fold inhibition at 72 h (P<0.05). Interferon gamma (IFNgamma) showed an inhibitory effect on DNA synthesis (1.3-fold; P<0.01). In UM-SCV-6 cells, both IL-10 and IL-13 showed inhibitory effects on deoxyuridine incorporation (1.3- and 1.4-fold at 48 h, respectively; P<0.001) that were even more pronounced at 72 h (2.4- and 2.5-fold respectively; P<0.001). IFNgamma caused a 3.6-fold inhibition of DNA synthesis by UM-SCV-6 cells at 72 h (P<0.001). Both TFGbeta and TNF alpha inhibited uridine incorporation (3.0- and 1.6-fold at 48 h, respectively; 2.7-fold at 72 h for both factors). GM-CSF inihibited DNA synthesis by UM-SCV-6 cells 1.3- 2.0-fold at 48 h and 72 h, respectively. In dose/response analyses, the effect of INF alpha on DNA synthesis was inhibitory in both cell lines at 48 h, while stimulatory effects were observed at 72 h. Electrophoretic analyses of DNA isolated from cells cultured in the presence or absence of different factors did not reveal DNA fragmentation. All cytokines, with the exception of IFN alpha, showed inhibitory effects on DNA synthesis by vulvar carcinoma cells. Of the factors studied, the recently described interleukins 10 and 13 showed potent inhibition of cell growth, encouraging further investigation on the molecular mechanisms of the observed inhibition. Apoptosis does not seem to be induced in the two vulvar carcinoma cell lines by any of the cytokines studied.

Measurement of activin B in human saliva and localization of activin subunits in rat salivary glands.

The present paper is the first report to demonstrate that measurable amounts of activin B are secreted into the saliva. The results show wide fluctuations in activin B concentrations during the menstrual cycle with peak values detected at the follicular phase. Estrogen replacement therapy was found to increase salivary activin B levels in post-menopausal subjects. The concentration in males was negligible. These data suggest that activin B concentrations in the human saliva may be under hormonal regulation and propose that salivary activin B measurements may prove useful in investigating the as yet less well defined local and/or physiological roles of activin B. The present paper reports, in addition, the immunohistochemical localization of activin/inhibin subunits in the duct systems of the rat submandibular, sublingual and parotid salivary glands.

Mechanisms of action of sex steroid hormones: basic concepts and clinical correlations.

The review deals with the clinically important aspects of the basic mechanisms of sex steroid hormones. Steroids can act through two basic mechanisms: genomic and non-genomic. The classical genomic action is mediated by specific intracellular receptors, whereas the primary target for the non-genomic one is the cell membrane. Many clinical symptoms seem to be mediated through the non-genomic route. Furthermore, membrane effects of steroid and other factors can interfere with the intranuclear receptor system inducing or repressing steroid-and receptor-specific genomic effects. These signalling pathways may lead to unexpected hormonal or anti-hormonal effects in patients treated with certain drugs. Steroid receptors (SRs) are members of a large family of nuclear transcription factors that regulate gene expression by binding to their cognate steroid ligands, to the specific enhancer sequences of DNA (steroid response elements) and to the basic transcription machinery. SRs are phosphoproteins, which are further phosphorylated after ligand binding. The role of phosphorylation in receptor transaction is complex and may not be uniform to all SRs. However, phosphorylation/dephosphorylation is believed to be a key event regulating the transcriptional activity of steroid receptors. SR activities can be affected by the amount of SR in the cell nuclei, which is modified by the rate of transcription and translation of the SR gene as well as by proteolysis of the SR protein. There is an auto- and heteroregulation of receptor levels. Some of the SRs appear to bind specific protease inhibitors and exhibit protease activity. The physiological significance of this weak proteolytic activity is not clear. Some SRs are expressed as two or more isoforms, which may have different effects on transcription. Receptor isoforms are different translation or transcription products of a single gene. Isoform A of the progesterone receptor is a truncated form of PR isoform B originating from the same gene, but it is able to suppress not only the gene enhancing activity of PR-B but also that of other steroid receptors. From the clinical point of view, it is important to note that the final hormonal effect in a target tissue is dependent on the cross talk between different nuclear steroid receptors and on expression of receptor isoforms.