Minoru Ueki

Effect of continuous combined therapy with vitamin K2 and vitamin D3 on bone mineral density and coagulofibrinolysis function in postmenopausal women

OBJECTIVES To investigate the therapeutic effect of combined use of vitamin K(2) and D(3) on vertebral bone mineral density in postmenopausal women with osteopenia and osteoporosis. SUBJECTS AND METHODS We enrolled 172 women with vertebral bone mineral density <0.98 g/cm(2) (osteopenia and osteoporosis) as measured by dual-energy X-ray absorptiometry. In this study, we employed the criteria for diagnosis of osteopenia and osteoporosis using dual energy X-ray absorptiometry proposed by the Japan Society of Bone Metabolism in 1996. Subjects were randomized into four groups (each having 43 subjects in vitamin K(2) therapy group, vitamin D(3) therapy group, vitamin K(2) and D(3) combined therapy group, or a control group receiving dietary therapy alone) and treated with respective agents for 2 years, with bone mineral density was measured prior to therapy and after 6, 12, 18, and 24 months of treatment. The bone metabolism markers analyzed were serum type 1 collagen carboxyterminal propeptide (P1CP), serum intact osteocalcin, and urinary pyridinoline. Tests of blood coagulation function consisted of measurement of activated partial thromboplastin time (APTT) and analysis of concentrations of antithrombin III (AT III), fibrinogen, and plasminogen. RESULTS Combined therapy with vitamin K(2) and D(3) for 24 months markedly increased bone mineral density (4.92 +/- 7.89%), while vitamin K(2) alone increased it only 0.135 +/- 5.44%. The bone markers measured, revealed stimulation of both bone formation and resorption activity. We observed an increase in coagulation and fibrinolytic activity that was within the normal range, suggesting that balance was maintained in the fibrinolysis-coagulation system. CONCLUSIONS Continuous combination therapy with vitamin K(2) and D(3) may be useful for increasing vertebral bone mass in postmenopausal women. Furthermore, the increase in coagulation function observed during this therapy was within the physiological range, and no adverse reactions were observed.

Correlation between vascular endothelial growth factor-C expression and invasion phenotype in cervical carcinomas

The correlation between vascular endothelial growth factor (VEGF)‐C gene expression and in vitro invasive activity and matrix metalloproteinase (MMP)‐2 or 9 gene expression and proteolytic activity in 11 cervical carcinoma cell lines, was investigated. Immunohistochemical expression of VEGF‐C in 52 cervical carcinoma tissues was also correlated with tumor aggressiveness with respect to clinicopathologic features, tumor vascularity, MMP‐2 expression and patient outcome. Expression of VEGF‐C mRNA differed remarkably among the cell lines and there was a statistical correlation between VEGF‐C gene expression and the number of invaded tumor cells (p = 0.0009) and MMP‐2 gene expression and activity (p < 0.05). Anti‐VEGF‐C antibody inhibited the invasive and proteolytic activity of tumor cells in a concentration‐dependent manner. VEGF‐C or MMP‐2 expression in clinical tissue samples was well correlated with depth of myometrial invasion, endometrial invasion, pelvic lymphnode metastasis and tumor vascularity (p < 0.05) and there was a close relation between VEGF‐C and MMP‐2 expression (p < 0.0001) in cervical carcinomas. Overall survival rates for 14 patients with strong VEGF‐C staining tumors were lower than those for 38 patients with weak VEGF‐C staining tumors (p = 0.0132) and VEGF‐C tissue status emerged as an independent prognostic parameter (p = 0.0232). These results suggest that VEGF‐C expression is closely related to invasion phenotype and affects the patients survival in cervical carcinomas.

Ovarian tumors associated with pregnancy

Objective: The clinical characteristics of ovarian tumors associated with pregnancy were investigated. Method: A series of 106 cases of ovarian tumor surgically resected during pregnancy was investigated with respect to incidence, clinicopathologic features and outcome. Results: Among 106 cases undergoing ovarian surgery, 31 (29.2%), 70 (66%) and five (4.7%) were diagnosed as physiologic, benign and malignant, respectively. The incidence of benign neoplastic tumor was 1:112 deliveries and that of malignant neoplastic tumor was 1:1684 deliveries. Dermoid cyst was the most common lesion found. Of the 70 benign tumors, 51 (72.9%) were greater than 8 cm in diameter and 55 (78.6%) were preoperatively diagnosed before the 10th gestational week; 44 (62.9%) were operated before the 15th gestational week. The spontaneous abortion rate in 80 cases followed up after surgery was only 10%, 61 patients (76.3%) progressing to full‐term delivery. Five malignant tumors included three epithelial carcinomas, one embryonal carcinoma and one dysgerminoma. Conclusion: Ovarian surgery in the first trimester for persistent or enlarging masses is important to obtain a final histologic diagnosis and rule out malignancy.

Vascular endothelial growth factor-C expression and invasive phenotype in ovarian carcinomas

Purpose: To investigate the biological correlation between vascular endothelial growth factor (VEGF)-C expression and invasive phenotype in ovarian carcinomas. Experimental Design: Gene and protein expression levels of VEGF-C in 10 ovarian carcinoma cell lines were correlated with invasive activity of the cells. The correlation between immunohistochemical expression of VEGF-C and tumor aggressiveness in 73 ovarian carcinomas was also examined with respect to clinicopathologic features and patient outcome. Results:VEGF-C gene and protein expression differed remarkably among the cell lines, and there was a statistical correlation among VEGF-C expression, in vitro invasive activity, and matrix metalloproteinase-2 (MMP-2) gene expression and its activity. Anti-VEGF-C and anti-MMP-2 antibodies inhibited the invasive activity of tumor cells. VEGF-C expression in clinical tissue samples was well correlated with clinical stages, retroperitoneal lymph node metastasis, MMP-2 expression, angiogenesis, lymphangiogenesis, and low apoptotic index (AI). The patients whose tumors had strong VEGF-C expression and low AI underwent a poorer prognosis than did those with weak VEGF-C expression and high AI. Conclusion: VEGF-C expression is closely related to invasive phenotype and affects the patients survival in ovarian carcinomas.

Histologic study of endometriosis and examination of lymphatic drainage in and from the uterus

Histologic study of endometriosis and investigation of lymphatic drainage in and out of the uterus were conducted to obtain more information on the histogenesis of endometriosis. Endometriosis is a disease originating from the normal endometrium, specifically from its basal layer. Internal endometriosis (adenomyosis) is caused mainly by direct invasion of the endometrium into the spaces located in the connective tissue of the muscle layer. Serous and ovarian types of endometriosis are frequently found together with adenomyosis. Endometrial fragments in vessels are detected in 4.5% of cases of endometriosis. Observation of serial sections proved that serous endometriosis is caused mainly by continuous or semicontinuous invasion of the tunica muscularis spaces primarily from adenomyosis lesions and partly from the endometrium. Confirmation of lymph flow into the ovary from the uterine body strongly suggests that ovarian endometriosis arises as a consequence of vascular (lymphatic) transport of endometrial fragments from adenomyosis lesions, serous endometriosis lesions, or the endometrium.

Vascular smooth muscle cell growth-promoting factor/F-spondin inhibits angiogenesis via the blockade of integrin αvβ3 on vascular endothelial cells

Vascular smooth muscle cell growth‐promoting factor (VSGP) was originally isolated from bovine ovarian follicular fluid as a stimulator of vascular smooth muscle cell proliferation. Homology searches indicate that bovine and human VSGPs are orthologs of rat F‐spondin. Here, we examined whether recombinant human VSGP/F‐spondin affected the biological activities of endothelial cells. VSGP/F‐spondin did not affect the proliferation of human umbilical vein endothelial cells (HUVECs); however, it did inhibit VEGF‐ or bFGF‐stimulated HUVEC migration. To clarify the mechanism of this inhibitory effect, we examined the adhesion of HUVECs to extracellular matrix proteins. VSGP/F‐spondin specifically inhibited the spreading of HUVECs on vitronectin via the functional blockade of integrin αvβ3. As a result, VSGP/F‐spondin inhibited the tyrosine phosphorylation of focal adhesion kinase (FAK) when HUVECs were plated on vitronectin. Moreover, VSGP/F‐spondin inhibited the activation of Akt when HUVECs on vitronectin were stimulated with VEGF. VSGP/F‐spondin inhibited tube formation by HUVECs in vitro and neovascularization in the rat cornea in vivo. These results indicate that VSGP/F‐spondin inhibits angiogenesis at least in part by the blockade of endothelial integrin αvβ3.

Gene expression of adhesion molecules and matrix metalloproteinases in endometriosis

Various types of cell adhesion molecules and matrix metalloproteinases (MMPs) seem to play an important role in the invasion process of endometriosis; however, limited investigation has focused on their gene expression in human peritoneal endometriotic lesions. A total of 63 endometriotic tissues were surgically obtained from 35 women with endometriosis, which included 43 pigmented and 20 non-pigmented lesions. Gene expression levels of E-cadherin, α- and β-catenin, MMP-2, MMP-9 and membrane-type 1 (MT1)-MMP in these endometriotic lesions were compared with those in normal eutopic endometrium obtained from 12 women without endometriosis. MMP-2, MMP-9 and MT1-MMP mRNA expression in pigmented lesions was significantly higher than that in normal endometrium (p < 0.05), whereas E-cadherin, α- and β-catenin mRNA expression was not suppressed in endometriotic lesions. There was a close correlation between MMP-2 or MT1-MMP and E-cadherin, α- or β-catenin gene expression in 63 endometriotic tissues examined (p < 0.01). Immunohistochemical expression of E-cadherin, α- and β-catenin in glandular epithelial cells was positive not only for all of seven cases with normal eutopic endometrium but also for 9 of 11 with ovarian endometriosis. MMP expression in ectopic endometrium was much greater than that in eutopic endometrium. These results suggest that endometriotic tissues expressing MMPs might be invasive and simultaneously possess cell-to-cell adhesion property in pelvic peritoneal foci.

Prevalence, incidence, and awareness in the treatment of menopausal urinary incontinence.

OBJECTIVES To investigate the prevalence of urinary incontinence and to evaluate the awareness of treatment in postmenopausal women. METHODS The study group was comprised of 3026 postmenopausal women consulting the outpatient clinic. One component of the urological questions was formulated to determine the voiding habits and presence or absence of urinary incontinence. The incontinent responders were questioned further regarding the nature of the urine loss to determine the severity of incontinence. RESULTS It was reported by 26.3% (795/3026) of the respondents that they currently had urinary incontinence. They were classified by types as follows: stress 64.9%, urge 18.6%, and mixed incontinence 7.3%. The incidence of women who desired medical treatment for incontinence was only 2.9% (87/3026), while the incidence of women who actually received medical treatment for urinary incontinence was 1.9% (56/3026). CONCLUSIONS We conclude that urinary incontinence was prevalent to a significant extent among postmenopausal women, but that few affected individuals in this group consulted the medical center desiring clinical treatment. The present study demonstrated that it is necessary to improve the awareness of postmenopausal women concerning the benefits of seeking treatment for urinary incontinence, in order to improve their quality of life.

Homeobox gene expression and mutation in cervical carcinoma cells.

An association between deregulation of homeobox (HOX) gene expression and oncogenic transformation has been recently reported in human tumors. In this study, we investigated HOX gene expression and mutation in cervical carcinoma cells. Using reverse transcription‐PCR, 11 human cervical carcinoma cell lines and 14 normal cervical tissue samples were examined for mRNA expression of the 39 class I HOX genes. DNA samples from 11 cell lines were tested for mutations in exons 1 and 2 of the HOXA10 and A13 genes using overlapping primer pairs which also cover intron 1 of these genes. HOXA1, B2, B4, C5, C10 and D13 genes were expressed in 8, 7, 9, 9, 9 and 11 of 11 cervical carcinoma cell lines, respectively, but not in any of the normal cervical tissues. HOXA9, A11, A13, B5, C4, D3 and D9 genes were expressed in all cell lines and normal tissues. In contrast, 13 of 39 HOX genes were silent in all materials examined. Single‐strand conforma‐tional polymorphism and sequence analysis revealed a C insertion after base 1042 and/or a G to C substitution at base 1113 in intron 1 of the HOXA13 gene in 4 of 11 cell lines, however, neither deletions nor mutations were detected in exons 1 and 2 of the HOXA10 and A13 genes. Our data suggest that the expression of HOXA1, B2, B4, C5, C10 and D13 genes might be involved in the process leading to the transformation of normal cervical cells. (Cancer Sci 2003; 94: 437–441)

Correlation between thymidine phosphorylase expression and invasion phenotype in cervical carcinoma cells

The correlation between thymidine phosphorylase (dThdPase) expression and invasion phenotype in human uterine cervical carcinoma cells was investigated using 10 cervical carcinoma cell lines. Semi‐quantitative reverse transcription‐polymerase chain reaction analysis was performed to investigate the mRNA levels of dThdPase and matrix metalloproteinase (MMP)‐2 with β‐actin coamplified as an internal standard. dThdPase protein expression levels were detected by highly sensitive enzyme‐linked immunosorbent assay. Tumor cell migration along a gradient of substratum‐bound fibronectin and invasion into reconstituted basement membrane were evaluated by haptotactic migration and invasion assay. Although dThdPase mRNA and protein expression levels differed remarkably among the cell lines, there was a statistical correlation between them (r = 0.743, p = 0.0139). dThdPase gene and protein expression levels were well correlated with the number of cells that migrated and invaded (p < 0.05). Moreover, there was a close correlation between MMP‐2 gene and dThdPase gene and protein expression levels (p < 0.05). Tumor cells that produce dThdPase may have a higher invasive and metastatic potential because of their capacity to pass through tissue barriers.