Expression of tumor-associated differentially expressed Gene-14 (TADG-14/KLK8) and its protein hK8 in uterine endometria and endometrial carcinomas.

To clarify the biological behavior of TADG-14/KLK8, we investigated TADG-14/KLK8 mRNA by semiquantitative RT-PCR and hK8 expression by immunohistochemistry using 37 normal endometria and 44 endometrial carcinoma tissues. TADG-14/KLK8 mRNA expression levels were significantly higher in proliferative compared to secretory phase endometria (p = 0.0143). Levels of TADG-14/KLK8 mRNA expression correlated with hK8 protein levels. hK8 was detected in 73.3% (11/15) of endometria with a significantly higher detection rate in the proliferative compared to secretory and atrophic phase endometria (p = 0.0002). High expression of hK8 was found in 61.4% of endometrial carcinomas compared to 35.1% of endometrial tissue samples (p = 0.0187). hK8 expression was significantly higher in stage I (p = 0.0433, 0.0038) and grade 1/2 (G1/2) of the tumors (p = 0.0195, 0.0044). We suggest that expression of TADG-14/KLK8may be regulated by sex steroid hormones in endometria. Our results indicate that elevated TADG-14/KLK8 expression is an early event in endometrial carcinogenesis, and may potentially serve as a useful early biomarker for the detection of endometrial carcinomas in menopausal women.

Cyclic and characteristic expression of phosphorylated Akt in human endometrium and decidual cells in vivo and in vitro.

BACKGROUND: Akt is activated by phosphorylation and plays an important role in cell survival and maintenance of structure. METHODS: We investigated whether phosphorylated Akt was characteristically expressed in human endometrium in vivo and whether insulin-like growth factor-I (IGF-I) can activate Akt using cultured decidualized human stromal cells in vitro, using immunohistochemistry and Western blotting analysis. RESULTS: The levels of phosphorylated Akt protein increased markedly in the decidual tissues from ectopic pregnancy. The expression of phosphorylated Akt protein in stromal cells increased with the decidualization. The decidual cells showed strong cytoplasmic staining for phosphorylated Akt. However, cultured decidualized human stromal cells diminished phosphorylated Akt expression compared to control cells. IGF-I administration to decidualized human stromal cells significantly recovered pAkt expression. The effect of IGF-I on decidualized human stromal cells was blocked by an inhibitor of phosphatidylinositol-3 kinase (PI3K) (LY294,002). These results suggest that IGF-I may activate Akt via PI3K in human endometrium and decidua. The expression of phosphorylated Akt in stromal cells was only detected in the functional layer, where tissue remodelling occurs during menstruation or implantation. CONCLUSIONS: Akt activation may be involved in cell survival and extracellular matrix remodelling in human endometrium and decidua.

Visual assessment of the mandibular cortex on panoramic radiographs to identify postmenopausal women with low bone mineral densities.

OBJECTIVE: The purpose of this study was to determine whether simple visual estimation of the mandibular inferior cortex width on panoramic radiographs is useful for identifying postmenopausal women with low skeletal bone mineral density (BMD). STUDY DESIGN: Panoramic radiographs were made on 100 women aged 50-84 years who also had BMD assessment of the lumbar spine and femoral neck by dual-energy x-ray absorptiometry. The panoramic images were examined twice by 4 observers to evaluate the thickness of the mandibular inferior cortex. RESULTS: Intraobserver agreement varied from moderate to almost perfect. Interobserver agreements varied from fair to almost perfect. Overall mean sensitivity, specificity and positive predictive value in identifying women with low BMD were 54.7%, 82.0%, and 72.5% in the lumbar spine and 51.8%, 82.2%, and 75.7% in the femoral neck, respectively. CONCLUSIONS: Our results suggest that simple visual estimation of the mandibular inferior cortex on panoramic radiographs may be useful for identifying postmenopausal women with low BMD.

Relative expression levels of Th1 and Th2 cytokine mRNA are independent prognostic factors in patients with ovarian cancer.

The aim of the present study was to examine mRNA expression levels of Th1 (TNF-alpha , IFN-gamma, and IL-12p40) and Th2 (IL-6 and IL-10) cytokines for any association with clinicopathological characteristics of epithelial ovarian cancer. mRNA was isolated, and cDNA prepared from 40 samples of epithelial ovarian cancers. Expression level of each cytokine mRNA was examined by the real-time PCR technique (GAPDH gene, internal control). Expression ratio (target gene/GAPDH) was used to evaluate gene expression. Results were analyzed against clinical stage, histological grade, and histological type. Prognostic significance of expression levels of each combination of Th1/Th2 values was assessed. Tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) expression levels were significantly higher in serous adenocarcinoma than in non-serous adenocarcinoma (p<0.05), but with no difference between individual cytokine mRNA expression levels and clinical stage or histological grade. Log-rank testing showed that high TNF-alpha mRNA expression (p=0.033) and the diameter of largest residual lesion at initial surgery (p=0.012) significantly correlate with longer survival in advanced stage (II/III/IV) ovarian carcinomas. In examining all combinations of Th1/Th2 expression values, the most significant association was between high IFN-gamma.IL-12p40/IL-6 expression levels and better prognosis in advanced stage (II/III/IV) ovarian carcinomas (p=0.004). In multivariate analysis, high IFN-gamma.IL-12p40/IL-6 expression (p=0.009) and the diameter of residual lesion (p=0.011) remained significantly associated with survival, whereas high TNF-alpha expression lost significance. In conclusion, Th1 and Th2 cytokines might play an important role in regulating the immune reaction in epithelial ovarian cancer cells. IFN-gamma.IL-12p40/IL-6 expression may be a useful prognostic molecular marker for patients with advanced ovarian cancer.

Relationship between self-reported periodontal status and skeletal bone mineral density in Japanese postmenopausal women.

OBJECTIVE: Several investigators have linked periodontal disease progression and low skeletal bone mineral density in postmenopausal women. However, little is known about whether self-reported periodontal status is the reflection of skeletal bone mineral density. We investigated whether self-reported poor periodontal status is associated with low skeletal bone mineral density in postmenopausal women. DESIGN: Relationships among self-reported periodontal status, number of teeth remaining, and bone mineral density of the lumbar spine and the femoral neck were evaluated in 253 Japanese postmenopausal women (mean +/- SD, 56.6 +/- 7.7) recruited from the patients who visited our clinic for bone mineral assessment between 1997 and 2003. Self-reported periodontal symptoms included gingival swelling, gingival bleeding, purulent discharge, and tooth mobility at the time of bone mineral assessment. RESULTS: Analysis of covariance adjusted for age, height, weight, years since menopause, duration of estrogen use, and regular oral care revealed that subjects without periodontal symptoms had significantly higher BMD of the lumbar spine than did those with periodontal symptoms (mean +/- SEM, 0.962 +/- 0.014 vs 0.921 +/- 0.013; P = 0.038); however, there were no significant differences in the number of remaining teeth and bone mineral density of the femoral neck between them. The odds of low spine bone mineral density in subjects with periodontal symptoms was 2.01 (95% CI = 1.15 to 3.50). CONCLUSION: Our results suggest that self-reported poor periodontal status may be associated with low bone mineral density of the lumbar spine in postmenopausal women.

Pregnancy rate, multiple pregnancy rate, and embryo quality: Clues for single blastocyst transfer from double blastocyst transfer in an unselected population.

Objective: Minimizing multiple pregnancy is a priority in assisted reproduction. As implantation rates are critical to success and reduce multiple pregnancy, we investigated whether blastocyst grade determined implantation rate following double blastocyst transfer in unselected cases. Materials and Methods: We studied 69 three-cleavage stage embryo transfers and 64 two-blastocyst transfers. Two blastocysts, or one when two blastocysts were not available, were transfered after evaluating the grade of blastocysts. The difference in pregnancy and implantation rates to patient age, the number of retrieved oocytes and grade of blastocysts were analyzed. Results: Blastocyst and grade 3AA rates per fertilized egg were 50.3% and 26.0%, respectively. Following two-blastocyst transfer, pregnancy rate per transfer, implantation rate per embryo, and multiple pregnancy rate per pregnancy were 39.1%, 26.5%, and 24.0%, respectively. Two-blastocyst transfer achieved implantation more often than three-cleavage-stage embryo transfer, but did not reduce multiple pregnancy. Pregnancy, implantation, and multiple pregnancy rates did not reflect maternal age. Higher pregnancy and implantation rates per transfer were attained for with six or more oocytes retrieved or transfer of two-blastocyst graded 3AA or higher especially when two or more blastocysts graded 3AA or higher are available, but the latter showed a high multiple pregnancy rate (38.5%). Conclusions: Single embryo transfer could be carried out when two or more blastocysts of grade 3AA or higher have been developed. (Reprod Med Biol 2005; 4: 153-160).

Prediction of outcomes of assisted reproduction treatment using the calcium ionophore-induced acrosome reaction.

BACKGROUND: Sperm concentration and motility are poor predictors of the outcome of intrauterine insemination (IUI), hysteroscopic intratubal insemination (HIT), or complete fertilization failure (CFF) in conventional IVF. We investigated whether the calcium ionophore-induced acrosome reaction (AR) constitutes an additional indicator of CFF and pregnancy that is independent of these semen parameters. METHODS: Infertile couples with no female factor (n=388) and women with tubal obstruction (n=32) were studied: IVF (n=133), ICSI (n=72), HIT (n=245) and IUI (n=61). The percentage of acrosome-reacted sperm in relation to viable sperm was calculated. Receiver operating characteristic curve and multiple logistic regression analyses were used to determine threshold values and the best predictor for CFF and pregnancy. RESULTS: Threshold values of AR for predicting CFF in IVF and pregnancy in IVF and HIT + IUI were 21, 26 and 22% respectively. These values were independent of the conventional semen analysis parameters. CFF was lower (2 versus 20%; P<0.01) and the pregnancy rate was higher (46 versus 24% P<0.05) for those with AR >21% in IVF. CFF and pregnancy rate in ICSI did not differ according to AR. Pregnancy rate was higher for those with an AR >22% for HIT + IUI (23 versus 11% P<0.01). CONCLUSIONS: Ionophore-induced AR appears to be a useful indicator in addition to routine semen analysis for selection of patients for treatment with appropriate assisted reproduction procedure.

Validation of dental panoramic radiography measures for identifying postmenopausal women with spinal osteoporosis.

OBJECTIVE: Measurements of mandibular inferior cortical shape and width detected on dental panoramic radiographs may be a useful screening tool for spinal osteoporosis in postmenopausal women. The purposes of this study were to clarify whether these measures are validated compared with simple screening tools based on questionnaires, such as the osteoporosis self-assessment tool (OST) and whether these measures can be used in postmenopausal women with histories of hysterectomy, oophorectomy, or estrogen use. SUBJECTS AND METHODS: We calculated the diagnostic performances of panoramic measurements and the OST for identifying women with spinal osteoporosis in both 159 healthy postmenopausal and 157 postmenopausal women with histories of hysterectomy, oophorectomy, or estrogen use. Spinal osteoporosis was defined as a bone mineral density T score of -2.5 or less at the lumbar spine. Cortical shape and width were evaluated on dental panoramic radiographs. Receiver operating characteristic curve analyses were used to determine the optimal cutoff thresholds for cortical width and the OST in healthy postmenopausal women. RESULTS: The sensitivity and specificity, respectively, for identifying women with spinal osteoporosis were 89.5% and 33.9% for cortical width, 86.8% and 57.8% for the OST, and 86.8% and 63.6% for cortical shape in healthy postmenopausal women. Sensitivity and specificity, respectively, were 92.5% and 35.0% for cortical width, 72.5% and 58.1% for the OST, and 80.0% and 64.1% for cortical shape in postmenopausal women with histories of hysterectomy, oophorectomy, or estrogen use. CONCLUSION: Dentists may be able to refer postmenopausal women with suspected spinal osteoporosis for bone densitometry on the basis of dental panoramic radiographs with diagnostic performance similar to that of osteoporosis screening tools based on questionnaires.

Apolipoprotein E phenotype affects the malondialdehyde-modified LDL concentration and forearm endothelial function in postmenopausal women.

OBJECTIVE: We investigated whether the apolipoprotein E (apo E) phenotype affects the serum concentration of malondialdehyde-modified low-density lipoprotein (MDA-LDL) or forearm endothelial function in postmenopausal women. PATIENTS AND MEASUREMENTS: Individuals were divided into three groups according to their apo E phenotype: E2 (E2/2 and E2/3, n = 12); E3 (E3/3, n = 71); and E4 (E3/4 and E4/4, n = 27). The serum concentrations of lipids and MDA-LDL were measured. Forearm blood flow during reactive hyperaemia and after sublingual nitroglycerin administration was measured by strain-gauge plethysmography. RESULTS: The serum concentrations of total and LDL cholesterol were significantly higher in the E4 group than in the E2 group (P < 0.05) or in the E3 group (P < 0.05). The serum apo B concentration was significantly higher in the E4 group than in the E2 group (P < 0.05). The serum concentrations of high density lipoprotein (HDL) cholesterol and nitrite/nitrate were significantly lower in the E4 group than in the E2 group (P < 0.05). Other lipid concentrations did not differ in the three groups. The serum MDA-LDL concentration was highest in the E4 group, and was lowest in the E2 group (E2: 91.1 +/- 6.9 IU/l, E3: 112.3 +/- 5.9 IU/l, E4: 128.8 +/- 9.9 IU/l; P < 0.05). The forearm blood flow response to reactive hyperaemia was lowest in the E4 group, and highest in the E2 group (E2: 52.2 +/- 5.8 ml/min per 100 ml tissue, E3: 40.7 +/- 1.7 ml/min per 100 ml tissue, E4: 33.4 +/- 2.4 ml/min per 100 ml tissue; P < 0.05). The forearm blood flow changes in response to nitroglycerine were similar between all three groups. CONCLUSIONS: The apo E phenotype affects the serum MDA-LDL concentration and forearm endothelial function in postmenopausal women.

Expression of human kallikrein 7 (hK7/SCCE) and its inhibitor antileukoprotease (ALP/SLPI) in uterine endocervical glands and in cervical adenocarcinomas.

To investigate the potential role of human kallikrein 7 (hK7/SCCE) and its inhibitor antileukoprotease (ALP/SLPI) in the development and progression of uterine cervical adenocarcinoma, we examined hK7 and ALP protein expression by immunohistochemistry in 70 cervical adenocarcinomas and 13 normal cervical tissues. Positive hK7 expression rates in normal endocervical glands and in cervical adenocarcinomas were 46.2 and 80%, respectively. A significantly higher hK7 expression rate was observed in cervical adenocarcinomas compared to normal endocervical glands (p=0.0099). In contrast, positive ALP detection rates in normal endocervical glands and in cervical adenocarcinomas were 100 and 15.7%, respectively. A significantly lower ALP detection rate was observed in cervical adenocarcinomas compared to normal endocervical glands (p<0.0001). There was a significant inverse correlation between hK7 and ALP expression status (p=0.0010). However, no statistically significant differences in hK7 or ALP expression status were found with respect to age, clinical stage, histological grade and lymph node metastasis status in cervical adenocarcinoma cases. Log-rank testing showed that advanced clinical stage and positive lymph node metastasis significantly correlated with poor patient survival (p=0.0005 and p<0.0001, respectively), whereas no correlation was found between hK7 or ALP expression and survival. These results suggest that increased expression of hK7 and decreased expression of ALP might play an important role in cervical adenocarcinoma development.

Effect of estrogen use on tooth retention, oral bone height, and oral bone porosity in Japanese postmenopausal women.

OBJECTIVE: Recent studies in the United States support the protective effect of estrogen use on tooth retention; however, little is known as to how estrogen promotes tooth retention. The aims of this study were to investigate the effects of estrogen use on tooth retention, oral bone height, and oral bone porosity in Japanese postmenopausal women and to clarify how estrogen promotes tooth retention. DESIGN: Relationships among the number of teeth remaining (total, anterior, and posterior teeth), oral bone height, oral bone porosity, bone mineral density of the lumbar spine and the femoral neck, estrogen use status, and the duration of estrogen use were evaluated in 330 Japanese postmenopausal women (mean age +/- SD, 56.8 +/- 7.6 y). RESULTS: Analysis of covariance adjusted for confounding variables revealed that estrogen users (66 women) tended to have more posterior teeth than did nonusers (264 women) (P = 0.065), although there were no significant differences in number of total (P = 0.196) and anterior (P = 0.751) teeth remaining, oral bone height (P = 0.970), oral bone porosity (P = 0.745), and bone mineral density of the lumbar spine (P = 0.459) and the femoral neck (P = 0.749) between estrogen users and nonusers. Multiple regression analysis showed that the duration of estrogen use was significantly associated with number of total (P = 0.019) and posterior (P = 0.007) teeth remaining, independent of age and oral bone height. CONCLUSION: Our results suggest that estrogen may promote tooth retention by strengthening the periodontal attachment surrounding the teeth, but not increasing oral bone height and not decreasing oral bone porosity.

Forearm endothelial function and bone mineral loss in postmenopausal women.

It is widely believed that the vasculature plays an important role in bone remodeling. We investigated the relationship between forearm endothelial function and bone mass in the lumbar spine in early postmenopausal women without a history of smoking or diabetes mellitus. We studied the forearm resistance artery endothelial function in 110 Japanese women-52 postmenopausal women with normal spinal bone mineral density (BMD), 36 postmenopausal women with osteopenia, and 22 osteoporotic postmenopausal women. Forearm blood flow (FBF) during reactive hyperemia and after sublingual nitroglycerin (NTG) administration was measured by strain-gauge plethysmography. BMD of the lumbar spine (L2-L4) was measured by dual-energy X-ray absorptiometry. After adjustment for age, body mass index, years since the start of menopause, and basal FBF, women with osteoporosis had a lower maximal FBF response to reactive hyperemia (28.4 +/- 3.8 mL/min per 100 mL tissue) than those with normal BMD (39.8 +/- 2.8 mL/min per 100mL tissue) or osteopenia (35.6 +/- 2.5 mL/min per 100mL tissue) (P = 0.029). A significant increase in serum angiotensin-converting enzyme (ACE) activity (P = 0.042) and a significant decrease in the serum concentrations of nitrite/nitrate (P = 0.041) were noted in osteoporotic women compared to women with normal BMD or osteopenia. The present findings suggest that postmenopausal women with low BMD, especially those with osteoporosis, have impaired endothelial function in the forearm resistance arteries.

Human kallikrein 8 (hK8/TADG-14) expression is associated with an early clinical stage and favorable prognosis in ovarian cancer.

The purpose of this study was to examine human kallikrein 8 (hK8/TADG-14) expression in epithelial ovarian tumors and to investigate the association of hK8 expression levels with patient survival. Human kallikrein 8 protein (hK8) expression was examined by immunohistochemistry in 74 ovarian adenocarcinomas and 6 normal ovaries. Results of immunostaining were correlated with clinicopathological variables and overall survival of the patients. Human kallikrein 8 gene (KLK8) mRNA expression was examined by semi-quantitative PCR in 35 ovarian tumors and 7 normal ovaries. Expression of hK8 was not detected on the surface epithelium of normal ovaries. In contrast, hK8 expression was detected in 51.4% (38/74) of carcinomas with a significantly higher detection rate of hK8 expression being observed in early stage disease compared to advanced stage disease (p=0.0192). Data analysis using the log-rank test showed hK8 expression correlated significantly with favorable patient survival (p=0.0328). Younger age (p=0.0008), early clinical stage (p<0.0001), and low histological grades of the tumors (p=0.0018) were also associated significantly with a favorable prognosis. In a multivariate model, age (p=0.0186) and clinical stage (p<0.0001) remained associated significantly with overall survival, whereas hK8 expression and histological grade lost their significance. There was significant relationship between the hK8 expression status and KLK8 mRNA expression levels (p=0.0304). Expression of hK8 is increased during the development of ovarian cancer and down-regulated during ovarian cancer progression. Expression of hK8 is a favorable prognostic marker in patients with ovarian cancer.

Tooth loss is associated with an increased risk of hypertension in postmenopausal women.

Tooth loss has been associated with an increased risk of vascular diseases such as coronary heart disease and cerebrovascular disease. Little is known whether hypertension is an important factor linking 2 phenomena in postmenopausal women. We compared an incidence of hypertension and traditional risk factors for vascular diseases between 2 age-matched groups: 67 postmenopausal women with missing teeth and 31 without missing teeth. In addition to blood pressure, serum concentration of total cholesterol, high- and low-density lipoprotein cholesterol and triglycerides, plasma angiotensin-converting enzyme activity, plasma angiotensin II concentration, plasma renin activity, and resting heart rate were measured as traditional risk factors for vascular diseases. Subjects without missing teeth had significantly lower diastolic blood pressure than did subjects with missing teeth (P=0.021). The former tended to have lower systolic blood pressure than did the latter (P=0.058). There were no significant differences in other variables between subjects with and without missing teeth. The odds ratio of having hypertension in subjects with missing teeth was 3.59 (95% confidence interval, 1.10 to 11.7) after adjustment of obesity, hypercholesterolemia, and hypertriglyceridemia. Our results suggest that hypertension may be an important factor linking tooth loss and an increased risk of vascular diseases in postmenopausal women.

Substitution of transdermal estradiol during oral estrogen-progestin therapy in postmenopausal women: effects on hypertriglyceridemia.

OBJECTIVE: We investigated effects of changing from oral estrogen to transdermal estradiol on the lipid and lipoprotein profile of postmenopausal women who developed hypertriglyceridemia (serum concentrations exceeding 150 mg/dL) during estrogen-progestin therapy. DESIGN: Sixty-one postmenopausal Japanese women receiving 0.625 mg of conjugated equine estrogen plus 2.5 mg of medroxyprogesterone acetate daily for 12 months had developed serum triglyceride concentrations exceeding 150 mg/dL after 12 months of treatment. Thirty-six of them, chosen randomly for study, were assigned at random to either a group that continued this oral regimen or another that changed to transdermal estradiol while continuing 2.5 mg of oral medroxyprogesterone acetate for the next 3 months (n = 18 for each). Blood lipids were compared between groups. RESULTS: Serum concentrations of triglyceride and very-low-density lipoprotein triglyceride decreased significantly after changing to transdermal estradiol (triglyceride, from 226.0 +/- 43.9 to 110.5 +/- 44.1 mg/dL, P < 0.01). No changes were seen in concentrations of low-density lipoprotein cholesterol or high-density lipoprotein cholesterol. CONCLUSION: Changing to transdermal estradiol may improve triglyceride metabolism in women who developed hypertriglyceridemia during oral estrogen-progestin therapy, with minimal effect on cholesterol profiles.

Human kallikrein gene 11 (KLK11) mRNA overexpression is associated with poor prognosis in patients with epithelial ovarian cancer.

PURPOSE: The purpose of this study was to examine expression levels of the human tissue kallikrein 11 gene (KLK11) in epithelial ovarian tumors and to identify the relationship between KLK11 expression and patient survival. EXPERIMENTAL DESIGN: KLK11 mRNA expression was examined by semiquantitative PCR in 64 epithelial ovarian tumors (7 adenomas, 6 low malignant potential tumors, and 51 adenocarcinomas) and in 10 normal ovaries. Semiquantitative PCR results were correlated with clinicopathologic variables and overall survival. cDNA from human normal tissues and tumor tissues was also analyzed. RESULTS: KLK11 mRNA expression was detected in various human cancer tissues including breast, lung, colon, prostate, pancreas, and ovarian carcinoma. The mean value of relative KLK11 expression ratio was significantly higher in ovarian tumor samples than in normal ovary samples (compared with normal samples: adenoma, P = 0.0006; low malignant potential tumor, P = 0.0049; and carcinoma, P < 0.0001). No statistically significant associations between KLK11 mRNA expression level and clinical stage, histological type, or histological grade were observed. The log-rank test showed that high KLK11 mRNA expression and advanced clinical stage significantly correlated with poor patient survival (P = 0.0185 and P = 0.0043, respectively). High KLK11 mRNA expression and clinical stage remained significantly associated with overall survival (P = 0.0225 and P = 0.0202, respectively) after multivariate analysis. CONCLUSIONS: KLK11 expression may play an important role in ovarian cancer development and act as an independent prognostic marker in ovarian cancer patients.

Mechanisms regulating the expression of indoleamine 2,3-dioxygenase during decidualization of human endometrium.

BACKGROUND: Expression of the tryptophan catabolizing enzyme, indoleamine 2,3-dioxygenase, in the mouse placenta has been shown to be critical in preventing immunological rejection of the fetal allograft. To clarify the physiological importance of indoleamine 2,3-dioxygenase in human pregnancy, we have studied how the expression of this enzyme changes during decidualization of human endometrium at both the cell and tissue level. METHODS AND RESULTS: The level of indoleamine 2,3-dioxygenase mRNA expression (determined by RT-PCR) was higher in decidual than in endometrial tissue. Uterine decidual tissue in ectopic pregnancy similarly showed increased mRNA expression. Immunohistochemistry demonstrated that indoleamine 2,3-dioxygenase protein immunoreactivity was found in glandular epithelium and in stromal cells. The intensity of this immunoreactivity was increased in decidualized tissue. In a cell culture model, the level of indoleamine 2,3-dioxygenase mRNA was suppressed specifically by progesterone-induced decidualization of isolated endometrial stromal cells. Indoleamine 2,3-dioxygenase protein abundance (determined by Western blot) was also decreased by progesterone-induced decidualization. However interferon-gamma, a potent stimulator of indoleamine 2,3-dioxygenase gene expression, increased the level of indoleamine 2,3-dioxygenase mRNA and protein in both non-decidualized and in decidualized cells. Indoleamine 2,3-dioxygenase activity (determined by measuring the concentration of tryptophan and its indoleamine 2,3-dioxygenase catabolite, kynurenine) was also decreased by progesterone-induced decidualization but enhanced following interferon-gamma treatment. Expression of other interferon-gamma inducible genes (STAT1 and tryptophanyl-tRNA synthetase) showed the same pattern as that of indoleamine 2,3-dioxygenase in tissue samples, but was not changed by decidualization in the cell culture model. CONCLUSIONS: These data suggest that despite suppression by progesterone, indoleamine 2,3-dioxygenase expression in endometrial stromal cells may increase during decidualization due to stimulation by interferon-gamma secreted by infiltrating leukocytes.

Indoleamine 2,3-dioxygenase: distribution and function in the developing human placenta.

Studies in mice have suggested that the placenta is protected from immune rejection by maternal T cells by means of localised indoleamine 2,3-dioxygenase dependent depletion of tryptophan. To determine whether such mechanisms might operate in the human placenta, we have studied the physiological importance of human placental indoleamine 2,3-dioxygenase immunohistochemically and functionally. Indoleamine 2,3-dioxygenase is detectable immunohistochemically from day 6 human blastocysts and thereafter throughout pregnancy in syncytiotrophoblasts, extravillous cytotrophoblasts and macrophages in the villous stroma and in the fetal membranes. Interferon-gamma added to villous explants markedly stimulates indoleamine 2,3-dioxygenase protein expression in macrophages. Indoleamine 2,3-dioxygenase-mediated tryptophan degradation in the first trimester villous and decidual tissue explants is stimulated by interferon-gamma and inhibited by 1-methyl-tryptophan (an inhibitor of indoleamine 2,3-dioxygenase). Peripheral blood mononuclear cell proliferation is controlled by indoleamine 2,3-dioxygenase-mediated tryptophan degradation. These results suggest the cellular basis of a mechanism present at the human maternal-fetal interface involved in regulating the maternal immune response to conceptus.

Human papillomavirus DNA status after loop excision for cervical intraepithelial neoplasia grade III - A prospective study.

The aims of the study were to investigate the relationship between human papillomavirus (HPV) DNA status and recurrence of cervical intraepithelial neoplasia (CIN) after loop excision (LEEP/LLETZ). Women (n=161) who underwent loop excision for CIN III and who were followed up prospectively for at least 4 years were the study cohort. Cervical smear cytology and testing for HPV DNA was performed at 3, 6 and 12 months prospectively and thereafter at intervals of 6-12 months, using the PCR method with a consensus primer targeting the L1 region. There has been no recurrence in 141 (81.6%) out of 161 subjects, while squamous intra-epithelial lesions (SIL) of low or high grade on cytology and CIN grade I-III on histology have been detected in 20 subjects. Prior to loop excision, HPV DNA was detected in 17 subjects who developed recurrence (9 had type 16, 2 each had types 18 and 52, and 1 each had types 31, 51, 58, and unknown). Within 3 months postoperatively, 12 (70.7%) subjects became negative for HPV, but 2 remained positive for the same type (1 each had types 16, 18), along with high-grade SIL on cytology, and CIN III on histology within 6 months, so repeat loop excision was performed. On the other hand, cytological findings were normalized in all transiently infected subjects within 18-36 months. Our results suggest that loop excision has improved HPV infection in many cases of CIN III and the persistent infection with a high-risk type of HPV is a predictor of the recurrence of CIN grade III.