Kohkichi Hata

Doppler ultrasound assessment of tumor vascularity in gynecologic disorders.

Real‐time two‐dimensional and pulsed‐wave Doppler ultrasonic examinations were performed on 8 normal volunteers and 97 patients with various gynecologic disorders; the objective was to assess uterine and tumor vascularities. Each arterial blood flow velocity wave‐form was classified into two types. The resistance indices of normal and abnormal flows were greater than .7 and less than .7, respectively. In normal volunteers, abnormal flows were nil. In 8 of 44 patients with benign tumors (18.2%), abnormal flows were evident and all proved to be cases of leiomyoma or adenomyosis. Doppler signals were not detected in 18 of 36 patients with cervical carcinoma (50%) and abnormal flows were noted in only 6 (16.7%). In all cases of endometrial carcinoma, ovarian carcinoma, and trophoblastic disease, typically abnormal flows were noted. Moreover, in most subjects a decrease in blood flows was observed after chemotherapy by anticancer drugs or irradiation. Therefore, Doppler ultrasound is a pertinent and noninvasive tool that can be used repeatedly for assessing the tumor vascularity in gynecologic disorders.

Hypoxia-inducible factor 1 alpha (HIF-1α) gene expression in human ovarian carcinoma

Abstract Hypoxia-inducible factor 1α (HIF-1α) that regulates genes involved in response to hypoxia and promotes neo-angiogenesis, is a transcriptional factor for vascular endothelial cell growth factor (VEGF). The aim of this study was to examine the expression of HIF-1α and VEGF gene expressions and their relation to angiogenesis, clinicopathologic variables and survival in the patient with human ovarian carcinoma. We retrospectively analyzed HIF-1α and VEGF gene expression levels using reverse transcriptase polymerase chain reaction (RT-PCR) in 60 ovarian carcinomas. Intratumoral microvessel density (IMD) was assessed by immunostaining endothelial cells, using anti-CD 31 antibody in frozen sections. The relationships between the expression level of these genes, IMD and clinicopathologic variables were evaluated by Students t-test and chi-square tests. Survival analysis was performed by Kaplan–Meier curves. HIF-1α or VEGF gene expression level was independent of age, clinical stage and histological subtype besides grade of tumor. There was no relationship between HIF-1α or VEGF gene expression level and IMD in all carcinomas (R=0.118 and 0.224, respectively). In addition, a weak association between HIF-1α and VEGF gene expression level was observed (R=0.300, P=0.020). The association between VEGF gene expression and IMD was observed (R=0.501, P=0.016). However, no association between IMD and HIF-1α gene expression was observed. Further, both HIF-1α and VEGF gene expression levels had no effect on survival in the patient with ovarian carcinoma. These results suggest that VEGF upregulated by HIF-1α gene may be involved in angiogenesis of some type of ovarian carcinoma, but the expression levels of both genes have no effect on survival in the patients with ovarian carcinoma.

Intratumoral peak systolic velocity as a new possible predictor for detection of adnexal malignancy

OBJECTIVE Our purpose was to evaluate whether intratumoral blood flow velocity measured by transvaginal Doppler ultrasonography would afford better discrimination between benign and malignant adnexal tumors. STUDY DESIGN One hundred two patients with adnexal tumors (72 benign, 30 malignant) were studied with transvaginal B-mode, color, and pulsed Doppler ultrasonography before surgery. RESULTS Sixty-five benign tumors (90.3%) had abnormal morphologic features suggesting malignancy, and blood flow was detected in 29 (40.3%). The peak systolic velocity was 13.1 +/- 9.1 cm/sec and the resistance index was 0.563 +/- 0.177. All malignant tumors had abnormal morphologic features and blood flow was detectable in all cases. The peak systolic velocity was 23.9 +/- 11.5 cm/sec and the resistance index was 0.488 +/- 0.118. When 16 cm/sec was considered as the cutoff value of peak systolic velocity, the sensitivity and specificity of peak systolic velocity in detecting malignant adnexal tumors were 83.3% and 91.6%, respectively. The sensitivity and specificity of the resistance index (cutoff value 0.72) was 93.3% and 68.1%, respectively. There was no significant difference in sensitivity between peak systolic velocity and resistance index. The specificity of the peak systolic velocity was significantly higher than that of the resistance index (p < 0.05). Moreover, the sensitivity of the peak systolic velocity did not significantly differ from that with transvaginal ultrasonography diagnosis (86.7%), and the specificity of the peak systolic velocity was significantly higher than that of transvaginal ultrasonography (69.4%). CONCLUSION These results suggest that peak systolic velocity obtained from intratumoral abnormal morphologic features provides a superior means for differentiating malignant from benign adnexal tumors.

Expression of Angiopoietin-1, Angiopoietin-2, and Tie2 Genes in Normal Ovary with Corpus luteum and in Ovarian Cancer

Objective: The recent discovery of angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2) has provided novel and important insights into the molecular mechanisms of blood vessel formation. Ang1 and Ang2 bind with similar affinity to the endothelial cell tyrosine kinase receptor Tie2. Our purpose was to assess the potential role of the Ang/Tie2 system in physiological and pathological angiogenesis in the ovary. Methods:Ang1, Ang2, and Tie2 gene expression in 14 normal ovaries with corpus luteum (CL) and in 19 cases of ovarian cancer were analyzed by polymerase chain reaction of RNA after reverse transcription. The level of each gene expression was presented by the relative yield of each gene to the β2-microglobulin gene, respectively. Furthermore, cellular distribution of Ang1 and Ang2 mRNA was examined by in situ hybridization, and localization of Tie2 was studied by immunochemistry. Results: The Ang1, Ang2, and Tie2 gene expression in normal ovary with CL ranged from 0.18 to 1.06 (median 0.54), 0.31–2.64 (median 1.01), and 0.10–0.47 (median 0.20), respectively. The expression of these same genes in ovarian cancer ranged from 0.06 to 0.75 (median 0.14), 0.69–1.59 (median 1.12), and 0.04–0.35 (median 0.15), respectively. Ang1 gene expression in normal ovary with CL was significantly higher than that in ovarian cancer (p = 0.0004). The gene expression levels of Ang2 and Tie2 were statistically the same in both groups. There was a significant correlation between Ang1 gene expression and Tie2 gene expression in normal ovary with CL (r = 0.619, p = 0.018). No such significant correlation was found in ovarian cancer. Moreover, Ang2 gene expression showed no significant correlation with the Tie2 gene expression either in normal ovary with CL or in ovarian cancer. Transcripts for Ang1 were observed in CL cells and endothelial cells around CL, and in tumor cells and endothelial cells at the periphery of tumor invasion. Ang2 transcripts were expressed in the same patterns. Tie2 expression was positive primarily in the endothelial cells around CL and in those at the periphery of tumor invasion. Conclusion: Our results indicate that there is a difference in the Ang/Tie2 gene expression between physiological and pathological angiogenesis in the ovary. This finding may aid in the development of new therapeutic interventions for ovarian cancer.

Gene Expression for Dihydropyrimidine Dehydrogenase and Thymidine Phosphorylase Influences Outcome in Epithelial Ovarian Cancer

PURPOSE Dihydropyrimidine dehydrogenase (DPD) is a pyrimidine salvage enzyme responsible for degradation of thymine, which is produced from thymidine by thymidine phosphorylase (TP). Our purpose was to determine whether DPD affects prognosis in patients with epithelial ovarian cancer and how the two enzymes may interact in such effects. PATIENTS AND METHODS DPD gene expression was analyzed by reverse transcription-polymerase chain reaction in 27 samples from normal ovaries and the 85 epithelial ovarian cancers previously studied with regard to TP gene expression. RESULTS DPD gene expression was significantly lower in epithelial ovarian cancers than in normal ovaries (P: <.0001), whereas TP gene expression and the ratio of TP to DPD gene expression (TP:DPD) were significantly higher in epithelial ovarian cancer (P: <.0001 for both). In patients with epithelial ovarian cancer, DPD gene expression and the TP:DPD ratio did not significantly correlate with any clinicopathologic factors. Patients with a high TP:DPD ratio (higher than the median) had significantly poorer outcomes than those with lower ratios (P: =.0002). The difference in survival between groups with high and low TP:DPD ratios was greater than the difference between groups with high and low TP gene expression. Multivariate analysis showed the TP:DPD ratio to be the independent prognostic factor (P: =.0002). In tumors with high TP gene expression, low DPD gene expression significantly correlated with poor survival (P: =. 04). CONCLUSION Downregulation of DPD gene expression may enhance the negative prognostic effect of high TP gene expression in patients with epithelial ovarian cancer. Certain newly available chemotherapeutic choices may take the TP:DPD ratio into consideration.

Maternal ophthalmic artery Doppler velocimetry in normotensive pregnancies and pregnancies complicated by hypertensive disorders.

OBJECTIVE Our objective was to compare maternal ophthalmic artery pulsatility index values in normotensive pregnancies and pregnancies complicated by hypertensive disorders. STUDY DESIGN The ophthalmic artery in 17 normotensive nonpregnant women, 29 normotensive pregnant women, 9 patients with mild preeclampsia, 6 with severe preeclampsia, 6 with transient hypertension, and 9 with chronic hypertension was studied with color Doppler flow imaging and pulsed Doppler ultrasonography. The mean arterial blood pressure and the ophthalmic artery pulsatility index were calculated in each group. RESULTS The pulsatility index (1.17 +/- 0.08) in severe preeclampsia was lowest among the groups p < 0.05), whereas that (2.92 +/- 0.59) in normotensive pregnant women was highest among the groups (p < 0.05). The pulsatility index (1.47 +/- 0.30) in mild preeclampsia was significantly lower than that (1.89 +/- 0.27) in transient hypertension (p < 0.05). There was no significant difference in pulsatility index between mild preeclampsia and chronic hypertension (1.69 +/- 0.49) or between transient hypertension and chronic hypertension. The pulsatility index inversely correlated well with the mean arterial blood pressure (R2 = 0.645, p < 0.0001). CONCLUSIONS These results suggest that the lower pulsatility index should be interpreted as orbital vascular vasodilation, indicating orbital hyperperfusion or hyperemia. Changes in pulsatility index in the ophthalmic artery may be indicative of similar changes in other cerebral vessels.

Prognostic value of overexpression of p53 in human ovarian carcinoma patients receiving cisplatin

A major obstacle to the treatment of ovarian carcinoma is intrinsic/acquired resistance to cisplatin-based chemotherapy. The clinical significance of p53 overexpression in patients with ovarian carcinoma is still controversial. The aim of this study was to investigate the independent prognostic significance of p53 overexpression in patients with ovarian carcinoma who are treated with cisplatin. We retrospectively examined the overexpression of p53 in primary ovarian carcinoma, and its association with chemotherapeutic efficacy. One hundred and thirty four ovarian carcinomas were surgically removed from patients who received adjuvant cisplatin-based chemotherapy. Immunohistochemical analysis of p53 was performed using a DO7 antibody against the p53 protein in 134 ovarian carcinomas. The significance of p53 in the prognosis of patients with ovarian carcinomas was also examined by a survival analysis of mortality follow-up data covering the period from 1988 to 2001. Thirty-three tumors (25%) exhibited p53 overexpression. Overexpression of p53 in grade 2/grade 3 tumors was significantly higher than that seen in grade 1 tumors (P=0.0088, 0.0229). Patients with tumors who also showed overexpression of p53 had a significantly inferior response to chemotherapy compared with the patients with p53-negative tumors (P=0.04). Cox regression analysis revealed that p53 overexpression was prognostic for poor disease outcome after adjustment for FIGO stage, grade and residual tumor. These findings suggest that overexpression of p53 in ovarian carcinoma is associated with unfavorable clinical outcome in patients treated with cisplatin-based chemotherapy. Therefore, detection of p53 overexpression using the DO7 antibody may be considered as a predictive marker of chemoresistance for cisplatin in patients with ovarian carcinoma.

Ovarian tumors of low malignant potential: transvaginal Doppler ultrasound features.

Twenty-nine patients with ovarian tumors were studied with transvaginal Doppler ultrasound before surgery. After surgery, pathological examination revealed that 26 tumors were benign and 3 were of low malignant potential (LMP). B-mode sonography, computed tomography, and magnetic resonance imaging showed no positive findings for malignancy in these 3 cases of LMP. Serum levels of the CA-125, tissue polypeptide antigen, and carcinoembryonic antigen were also within the normal range. Blood flow velocity waveforms were evaluated by the calculation of the resistance index (RI). There was a significant difference between the RI value (0.818 +/- 0.223) in benign tumors and that (0.418 +/- 0.072) in LMP (P less than 0.01). When the 0.56 (mean of LMP tumor RI value + 2SD) was considered as the cutoff value of RI, the sensitivity was 100% and the specificity was 88.5%. Transvaginal Doppler ultrasound provides a useful diagnostic information for the differentiation of benign and LMP ovarian tumors before surgery.

Clinical significance of interleukin-1 receptor antagonist in patients with cervical carcinoma

OBJECTIVE Our purpose was to determine the clinical significance of interleukin-1 receptor antagonist (IL-1ra), which is an endogenous inhibitor cytokine of IL-1, in patients with cervical carcinoma. METHODS Tissue IL-1ra expression and serum IL-1ra level were examined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and immunohistochemistry in normal controls and patients with cervical carcinoma. RESULTS Tissue IL-1ra protein level by ELISA was significantly higher in squamous cell carcinoma (n = 9) than in the normal cervix (n = 7) and adenocarcinoma (n = 3). Western blotting confirmed the main presence of intracellular IL-1ra type 1 in squamous cell carcinoma. Immunohistochemistry demonstrated significant IL-1ra expression only in tumor cells of squamous cell carcinoma. Elevation of serum IL-1ra level was found in patients with squamous cell carcinoma (n = 38) compared to normal women (n = 13), but not in patients with adenocarcinoma (n = 9). Although serum IL-1ra level did not correlate with clinical stage or any other tumor marker, high serum IL-1ra level was associated with pelvic lymph node metastasis and poor prognosis in patients with squamous cell carcinoma. On the other hand, these results were not obtained in patients with cervical adenocarcinoma. CONCLUSION IL-1ra may play important roles in local and general malignant behaviors in patients with cervical squamous cell carcinoma, and measurement of serum IL-1ra level may be useful in predicting patient survival.

Three-dimensional ultrasonographic assessment of the umbilical cord during the 2nd and 3rd trimesters of pregnancy

Objective: To evaluate the umbilical cord and its abnormalities by use of three-dimensional ultrasonography with a specially developed abdominal three-dimensional transducer. Patients and Methods: Ninety-five pregnancies (92 normal, 2 with hydrops fetalis, and 1 with omphalocele) from 14 to 40 weeks of gestation were studied with a specially developed abdominal three-dimensional transducer (3.5 MHz). This system can provide conventional two-dimensional ultrasonography images and can also generate within seconds high-quality three-dimensional images in the surface and transparent mode with no need for an external workstation. A proportion of the umbilical cords (coiled or noncoiled free loop, abdominal insertion, and placental insertion) visualized at each gestational age interval is presented. Results: The proportion of the umbilical cords visualized during pregnancy except for between 24 and 27 weeks of gestation was about 70% (range 64–83%). Optimal visualization of the umbilical cord was achieved between 24 and 27 weeks of gestation. During this period it was possible to adequately depict the umbilical cord in 93% of the cases. The proportions of the noncoiled umbilical cord depicted during pregnancy ranged from 8 to 45%. The detection rate of abdominal insertion of the umbilical cord visualized at 14–19 weeks was 44%; the detection rate decreased thereafter. Placental insertion of the umbilical cord could not be identified after 28 weeks of gestation. In 2 cases with hydrops fetalis, edematous umbilical cord was evident. In 1 fetus, omphalocele was clearly depicted. Conclusions: The new three-dimensional ultrasound technology generates within seconds high-quality three-dimensional images of the umbilical cord, although limitation of viewing direction exists. These results suggest that the new three-dimensional ultrasonography has the potential to be a supplement to two-dimensional ultrasonography and might be useful in identifying abnormal umbilical cords in utero.