Measurement of endometrial thickness in premenopausal women in office gynecology.

Purpose: To define the median endometrial thickness (ET) in office gynecology is thought to be important for clinical practice. However, there are few reports about ET that have included the general female population on a large scale. The median ET was determined prospectively in premenopausal women who attended office gynecology for cervical cancer screening. Methods: In total, 849 women were enrolled. The median ET was determined by using transvaginal ultrasound and the relationships between the ET and various clinical factors were analyzed. Results: The participants' median age was 38.5 years. The median ET was 8.6 mm (90% and 95% quantiles: 13.8 and 15.8 mm). The ET was not related to their age, symptoms, obstetric history, geographical location, or risk factors for endometrial cancer. In the women with a menstrual cycle length of 28-30 days, the ET was 7 mm on days 1-6, but it increased from 5.4 mm immediately after menstruation (day 7 or 8) to 9.2 mm on days 13-14. Subsequently, the ET increased further to 11.1 mm on day 18. Conclusion: In all the women, the upper limit of the ET was 13.8 mm and 15.8 mm in the 90% and 95% quantile, respectively, in office gynecology.

Allogeneic intrabone marrow-bone marrow transplantation plus donor lymphocyte infusion suppresses growth of colon cancer cells implanted in skin and liver of rats.

We have recently found that allogeneic intrabone marrow-bone marrow transplantation (IBM-BMT) + donor lymphocyte infusion (DLI) using CD4(+) cell-depleted spleen cells (CD4(-) cells) can prevent graft-versus-host disease (GvHD) but suppress tumor growth (Meth A: fibrosarcoma) in mice. In the present study, we show that allogeneic IBM-BMT + DLI using CD4(-) cells also has suppressive effects on the growth of colon cancer cells implanted not only in the skin but also in the liver of rats. First, we examined the effects of allogeneic IBM-BMT + DLI on the subcutaneously inoculated ACL-15 (rat colon cancer cell line). Lethally irradiated Fischer rats (F344 rats) were transplanted with T-cell-depleted bone marrow cells (BMCs) from Brown Norway (BN) rats. Simultaneously, DLI was performed using whole spleen cells (whole cells), CD4(+) cell-depleted spleen cells (CD4(-) cells) or CD8(+) cell-depleted spleen cells (CD8(-) cells) of BN rats. Although allogeneic IBM-BMT + DLI suppressed tumor growth, a considerable number of rats treated with allogeneic IBM-BMT + DLI using whole cells or CD8(-) cells died due to GvHD. In contrast, allogeneic IBM-BMT + DLI using CD4(-) cells also suppressed tumor growth, but there was no GvHD. Based on these findings, we next examined the effects of allogeneic IBM-BMT + DLI using CD4(-) cells on the cancer cells implanted in the liver. Allogeneic IBM-BMT + DLI using CD4(-) cells via the portal vein significantly prolonged the survival. These results suggest that allogeneic IBM-BMT + DLI using CD4(-) cells could become a new strategy for the treatment of solid tumors.

Effects of prepubertal indole-3-carbinol treatment on development of N-methyl-N-nitrosourea-induced mammary carcinomas in female Sprague-Dawley rats.

BACKGROUND: Indole-3-carbinol (I3C), which is present in cruciferous vegetables, has been shown to prevent the development of mammary cancer when administered to adult animals. However, no studies have been reported on the effects of prepubertal short-term I3C treatment on N-methyl-N-nitrosourea (MNU)-induced mammary carcinogenesis. MATERIALS AND METHODS: Prepubertal female Sprague-Dawley rats were administered the vehicle (Group 1) or I3C (Group 2, 250 mg/kg/day at 15 and 16 days of age; Group 3, 50 mg/kg/day at 15 and 16 days of age; Group 4, 50 mg/kg/day at 15, 16, 29 and 30 days of age; Group 5, 50 mg/kg/day at 29 and 30 days of age). All rats were administered 50 mg/kg MNU at 22 days of age. Rats were sacrificed at 34 weeks of age or when their largest mammary tumor reached a diameter of > or =1 cm. Body weight gain, vaginal opening, estrous cyclicity and mammary carcinogenesis were compared between the groups. RESULTS: Rats administered 250 mg/kg I3C exhibited acute toxicity, and 40% of that group died soon after administration of I3C. There was no significant difference in body weight and relative uterine-ovarian weight of surviving rats between groups at the end of the experiment. However, rats from Group 2 and Group 3 exhibited earlier vaginal opening and prolonged estrous cyclicity, respectively. I3C treatment before and after MNU administration (Group 4) tended to reduce mammary carcinoma incidence (percentage of mammary carcinomas with a diameter of > or =1 cm) and multiplicity (number of all-sized mammary carcinomas per rat), and prolonged the latency (time from MNU administration to point when mammary tumors grew to a diameter of > or =1 cm) compared with the vehicle (control) group. Mammary carcinogenesis was not altered by other I3C treatments. CONCLUSION: Prepubertal I3C treatment before and after carcinogen exposure appeared to provide an insignificant protection against MNU-induced mammary carcinogenesis.

Parous mammary glands exhibit distinct alterations in gene expression and proliferation responsiveness to carcinogenic stimuli in Lewis rats.

Early full-term pregnancy affords lifetime protection against the development of breast cancer. Parity-induced protection can be reproduced in a carcinogen-induced rat mammary carcinoma model, but the molecular mechanisms of this protection against carcinogenic stimuli in rat mammary glands have not been fully characterized. To gain a better understanding of these molecular mechanisms, we used an oligonucleotide microarray to examine gene expression in parous and age-matched virgin (AMV) mammary glands of Lewis rats before and after carcinogen (N-methyl-N-nitrosourea; MNU) treatment. Parous mammary glands before MNU treatment showed up-regulation of multiple differentiation-related genes, such as whey acidic protein (Wap), casein beta (Csn2), casein gamma (Csng), lipopolysaccharide binding protein (Lbp), secreted phosphoprotein 1 (Spp1) and glycosylation-dependent cell adhesion molecule 1 (Glycam1). Also, parous mammary glands before MNU treatment exhibited down-regulation of growth-related genes such as regenerating islet-derived 3 alpha (Reg3a), mesothelin (Msln), insulin-like growth factor 2 (Igf2) and insulin-like growth factor binding protein 4 (Igfbp4). After MNU treatment, AMV mammary glands exhibited up-regulation of growth-related genes, such as Msln, cell division cycle 2 homolog A (Cdc2a), Igf2, Igfbp4, stathmin 1 (Stmn1) and homeobox, msh-like 1 (Msx1), whereas expression of these genes remained low in parous mammary glands. AMV mammary glands also exhibited marked up-regulation of Cdc2a and Stmn1 in response to MNU. After MNU treatment, the PCNA labeling index increased significantly in AMV mammary epithelial cells (13.7+/-1.1%), but remained low in parous mammary glands (3.6+/-0.4%). The response of AMV mammary glands to carcinogenic stimuli includes up-regulation of growth-related genes and increased cell proliferation. The lack of a similar response in parous mammary glands may explain parity-induced protection against mammary tumor development.

Diagnostic cytopathology in type II angiosarcoma of the breast: a case report.

BACKGROUND: The rare occurrence of angiosarcoma of the breast is reflected in limited descriptions of fine needle aspiration (FNA) cytomorphologic findings in this neoplasm. We present a case of angiosarcoma of the breast and discuss the pitfalls in diagnostic cytopathology that can potentially lead to incorrect diagnoses in such cases. CASE: A 45-year-old woman presented with a 2-year history of a right-sided breast mass. FNA cytology revealed a hypocellular smear composed of cohesive ductal epithelial cells; isolated or loosely arranged, round to spindle-shaped fibroblastlike cells; and projectile growths of round, oval and polygonal cells on loose tangles of connective tissue. The background was hemorrhagic, with scattered foam cells. The overall cytologic diagnosis was inconclusive but suggested phyllodes tumor (of borderline malignancy). Excisional biopsy was performed, followed by simple mastectomy. Histologic features were consistent with angiosarcoma, a diagnosis that was supported by immunohistochemical studies. CONCLUSION: On FNA smear, 49.1% of isolated atypical cells were positive for the endothelial marker CD34; however, cytomorphologic appearance of these cells resembled that of CD34-negative active mesenchymal cells. Angiosarcoma rarely occurs in the breast, and a definitive diagnosis is difficult based on cytologic examination of hypocellular smears alone. However, the presence of atypical round, oval and polygonal cells on loose tangles of connective tissue, suggesting sheared vasculature, can be an important diagnostic feature.

Correlation of telomerase activity and telomere length to chemosensitivity.

Telomerase, which is selectively expressed in germline or cancer cells, is a ribonucleoprotein polymerase that contains an integral RNA with a short template element that can compensate telomeric loss by synthesizing TTAGGG repeats at chromosome ends. Telomeres appear to be critical for the integrity of chromosomes, stabilizing them from exonucleolytic degradation, preventing chromosome-to-chromosome fusions, and determining the maximum replicative capacity of cells. During the past decade, the roles of telomere length and telomerase activity have been investigated extensively in a variety of benign and malignant tumors of human origin, and stronger telomerase activity has been observed in more advanced tumors. Generally, the acquisition of telomerase activity in cancer cells is rather universal, which suggests that telomerase inhibition as a novel and potentially selective target for therapeutic intervention. Although a telomerase-specific inhibitor has not been found yet, the possible effect of anticancer agents on telomerase inhibition or the alteration of telomere length has been proposed. Recent development of TRAP assay not only increased the sensitivity but also allowed fast and efficient detection of telomerase activity. Technical aspects of this assay using self-established internal standard and nonradioisotopic detection method are addressed in this report. In addition, an overview of how to determine the telomere length is described.

Dietary factors modifying breast cancer risk and relation to time of intake.

Multiple factors contribute to the development of human breast cancer. However, environmental factors, especially dietary factors, appear to have the greatest effects. Evidence obtained in epidemiological studies has been corroborated by laboratory findings. Dietary components strongly associated with breast cancer include fat and phytochemicals. A diet high in n-3 polyunsaturated fatty acid (PUFA) or monounsaturated fatty acid (MUFA) and low in n-6 PUFA is protective against breast cancer. Some phytochemicals present in fruits and vegetables are also protective. Time of intake appears to be important: lifetime protection may be achieved if one is exposed to a dietary factor that lowers breast cancer risk early in life. Synergistic and antisynergistic interactions between dietary factors can modify breast cancer risk. The available evidence suggests that breast cancer risk can be reduced by early dietary intervention.

Conjugated docosahexaenoic acid suppresses KPL-1 human breast cancer cell growth in vitro and in vivo: potential mechanisms of action.

INTRODUCTION: The present study was conducted to examine the effect of conjugated docosahexaenoic acid (CDHA) on cell growth, cell cycle progression, mode of cell death, and expression of cell cycle regulatory and/or apoptosis-related proteins in KPL-1 human breast cancer cell line. This effect of CDHA was compared with that of docosahexaenoic acid (DHA). METHODS: KPL-1 cell growth was assessed by colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay; cell cycle progression and mode of cell death were examined by flow cytometry; and levels of expression of p53, p21Cip1/Waf1, cyclin D1, Bax, and Bcl-2 proteins were examined by Western blotting analysis. In vivo tumor growth was examined by injecting KPL-1 cells subcutaneously into the area of the right thoracic mammary fat pad of female athymic mice fed a CDHA diet. RESULTS: CDHA inhibited KPL-1 cells more effectively than did DHA (50% inhibitory concentration for 72 hours: 97 micromol/l and 270 micromol/l, respectively). With both CDHA and DHA growth inhibition was due to apoptosis, as indicated by the appearance of a sub-G1 fraction. The apoptosis cascade involved downregulation of Bcl-2 protein; Bax expression was unchanged. Cell cycle progression was due to G0/G1 arrest, which involved increased expression of p53 and p21Cip1/Waf1, and decreased expression of cyclin D1. CDHA modulated cell cycle regulatory proteins and apoptosis-related proteins in a manner similar to that of parent DHA. In the athymic mouse system 1.0% dietary CDHA, but not 0.2%, significantly suppressed growth of KPL-1 tumor cells; CDHA tended to decrease regional lymph node metastasis in a dose dependent manner. CONCLUSION: CDHA inhibited growth of KPL-1 human breast cancer cells in vitro more effectively than did DHA. The mechanisms of action involved modulation of apoptosis cascade and cell cycle progression. Dietary CDHA at 1.0% suppressed KPL-1 cell growth in the athymic mouse system.

Perillyl alcohol inhibits human breast cancer cell growth in vitro and in vivo.

The effect of monoterpene perillyl alcohol (POH) on cell growth, cell cycle progression, and expression of cell cycle-regulatory proteins in estrogen receptor (ER)-positive (KPL-1 and MCF-7) and ER-negative (MKL-F and MDA-MB-231) human breast cancer cell lines was examined. POH inhibited cell proliferation in a dose-dependent manner in all cell lines tested. POH at a dose of 500 micro M had a cytostatic effect, in which growth inhibition was due to accumulation of cells in G1-phase. Cell cycle progression was preceded by a decrease in G1 cyclins (cyclin D1 and E), followed by an increase in p21(Cip1/Waf1) and a decrease in proliferating cell nuclear antigen level. Levels of p53 and cyclin A were unchanged. POH at a dose of 75 mg/kg administered intraperitoneally three times a week throughout the entire 6-week experimental period suppressed orthotopically transplanted KPL-1 tumor cell growth and regional lymph node metastasis in a nude mouse system. POH inhibited both ER-positive and -negative human breast cancer cell growth in vitro, and suppressed growth and metastasis in vivo.

Massive telomere loss and telomerase RNA expression in dexamethasone-induced apoptosis in mouse thymocytes.

Apoptosis is a well-recognized process of cell death occurring under several physiological and pathological conditions and represents the principal mechanism involved in cell selection in the thymus. Glucocorticoids are well known to stimulate apoptosis in rat thymocytes. However, it is unclear whether the same changes occur after in vivo glucocorticoid treatment in mice. Chromosomal stability and cell viability require a proficient telomeric end-capping function. Cells with critical telomere shortening and telomerase dysfunction undergo increased apoptosis. In turn, the change in telomere function in cells undergoing apoptosis is not fully characterized. In order to investigate this, we studied the changes in thymocytes after dexamethasone administration in BALB/c mice. The loss of normal thymocytes coincided with the appearance of small dense cells with characteristic features of apoptosis including condensed chromatin, internucleosomal DNA cleavage, and a "hypodiploid" peak on flow cytometry, which suggested that dexamethasone-induced thymocyte apoptosis in BALB/c mice could be considered a well-defined experimental model for studying apoptotic processes. Dexamethasone-treated thymocytes exhibited rapid and dynamic loss of telomeric sequences and up-regulation of telomerase RNA as an early event in the apoptotic process. Telomerase activity was unchanged in this event. Thereafter, telomere gain associated with an increase in telomerase activity occurred in the regenerative process of the thymus. These results suggest a role of telomere loss and up-regulation of telomerase RNA as key apoptosis sensors.

Caspase-3 inhibitor transiently delays inherited retinal degeneration in C3H mice carrying the rd gene.

BACKGROUND: The effect of a caspase-3 inhibitor on retinal degeneration in C3H mice carrying the rd gene, a mutation of a rod-specific phosphodiesterase, was investigated. METHODS: A quantity of 2 mg/kg of Ac-DEVD-CHO, as inhibitor, was injected intraperitoneally every other day from 8 days of age, and retinal damage was compared with that in saline-treated C3H mice at 13 days (1 day after the third treatment) and 17 days of age (1 day after the fifth treatment). Retina of ICR mice not carrying rd gene was also evaluated under the same protocol. The efficacy of Ac-DEVD-CHO was evaluated based on total retinal thickness and outer retinal thickness (thickness of outer nuclear layer and photoreceptor layer). An apoptotic index and a cell proliferation index for the photoreceptor cells, at 13 days of age, were calculated based on terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling (TUNEL) and proliferating cell nuclear antigen (PCNA) labeling, respectively. RESULTS: At 13 days of age, total and outer retinal thickness in saline-treated C3H mice were 140.3 microm and 37.5 microm, compared with 160.4 microm and 49.5 microm, respectively, in Ac-DEVD-CHO-treated C3H mice ( P<0.01, respectively). In ICR mice, total and outer retinal thickness were 182.1 microm and 90.9 microm, respectively, in saline-treated mice and 183.8 microm and 89.6 microm in Ac-DEVD-CHO-treated mice (not significant). At this time, the TUNEL index was 23.52 cells/10(4) microm (2) of outer nuclear layer in saline-treated C3H mice; Ac-DEVD-CHO treatment significantly reduced this value to 18.73 cells/10(4) microm(2) ( P<0.05). The TUNEL index in saline- and Ac-DEVD-CHO-treated ICR mice was 0.59 cells/10(4) microm(2) and 0.80 cells/10(4) microm(2), respectively (not significant); Ac-DEVD-CHO treatment had no influence on normally developing retina. The PCNA index was not affected by Ac-DEVD-CHO-treatment. However, at 17 days of age, Ac-DEVD-CHO treatment did not ameliorate retinal degeneration. CONCLUSIONS: The caspase-3 inhibitor was transiently effective in delaying retinal degeneration through inhibition of the apoptosis of photoreceptor cells in rd gene-carrying mice. The use of caspase-3 inhibitors may have therapeutic applications in the treatment of human retinal degeneration.