Takao Fukaya

Statin Attenuates Increase in C-Reactive Protein During Estrogen Replacement Therapy in Postmenopausal Women

To the Editor: In their interesting study, Koh et al1 demonstrated that 0.625 mg of conjugated equine estrogen (CEE) daily for 6 weeks caused an increase in plasma C-reactive protein (CRP), whereas the combination of simvastatin 10 mg daily with CEE for 6 weeks reduces the estrogen-induced increase in CRP. The antiinflammatory effect of HMG-CoA reductase inhibitors (statins) may reduce the estrogen-induced increase in plasma concentrations of CRP. The Heart and Estrogen/Progestin Replacement Study (HERS) demonstrated that estrogen and progestin therapy did not reduce the overall rate of coronary events in postmenopausal women with established coronary disease.2 Because elevated CRP may be associated with plaque destabilization and rupture, a proinflammatory effect of estrogen might explain the increased number of cardiovascular events demonstrated in women with existing cardiovascular disease during the first year of the HERS trial. Accordingly, a combination of …

Superoxide dismutase in normal cycling human ovaries: immunohistochemical localization and characterization

OBJECTIVE To investigate the expression of manganese (Mn) and copper-zinc (Cu,Zn) superoxide dismutase (SOD) in normal cycling human ovaries throughout the menstrual cycle. DESIGN Descriptive, controlled study. SETTING Tohoku University School of Medicine. PATIENT(S) Twenty-four normal cycling human ovaries were obtained from patients who underwent oophorectomy and hysterectomy for squamous cell carcinoma of the uterine cervix. INTERVENTION(S) Immunohistochemistry for Mn-SOD and Cu,Zn-SOD. MAIN OUTCOME MEASURE(S) Immunostaining. RESULT(S) In the follicular stage, Mn-SOD immunoreactivity was detected in granulosa and theca interna cells of steroid-producing follicles, that is, preantral, nondominant, dominant, and atretic follicles, whereas Cu,Zn-SOD was detected in theca interna cells of these follicles and in granulosa cells of dominant follicles. In the luteal stage, immunoreactivity for Mn-SOD and Cu,Zn-SOD was observed in both luteinized granulosa and theca cells of the functioning corpus luteum. In the early degenerating corpus luteum, both Mn-SOD and Cu,Zn-SOD were positive in steroid-producing luteinized theca cells. Mn-SOD immunoreactivity was also detected in nonsteroid-producing luteinized granulosa cells and macrophages. CONCLUSION(S) Our results suggest that the expression of Mn-SOD and Cu,Zn-SOD closely correlates with steroidogenesis in the human ovary. In addition, Mn-SOD may play an important role in the process of luteal regression.

Melatonin protects fetal rat brain against oxidative mitochondrial damage.

Our objective was to investigate the effects of melatonin on the free radical‐induced oxidative damage to mitochondria in fetal rat brain. Female Wistar rats on day 19 of pregnancy were used. Melatonin (10 mg/kg) or vehicle (control) was injected intraperitoneally 60 min prior to laparotomy for removal of the fetuses. The mitochondrial fraction was isolated from the fetal rat brain of each group. Superoxide dismutase (SOD) and glutathione peroxidase (GSH‐Px) activities were measured. As indicators of mitochondrial respiratory activity, we determined the respiratory control index (RCI) and the adenosine 5‐diphosphate/oxygen (ADP/O) ratio in the presence and absence of 2.5 μM hypoxanthine and 0.02 units/mL xanthine oxidase. Mitochondrial lipid peroxidation was determined by measuring the concentration of thiobarbituric acid reactive substances in fetal brain mitochondria in the presence or absence of 2.5 μM hypoxanthine, 0.02 units/mL xanthine oxidase, and 50 μM FeSO4. The free radical‐induced rates of inhibition of mitochondrial RCI and the ADP/O ratio were both significantly lower in the fetal rat brains treated with melatonin compared with those of the controls (RCI, 44.25±15.02% vs. 25.18±5.86%, P<0.01; ADP/O ratio, 50.74±23.05% vs. 13.90±7.80%, P<0.001). The mitochondrial lipid peroxidation induced by free radicals was significantly reduced in the melatonin‐treated group compared with the controls (484.2±147.2% vs. 337.6±61.0%, P<0.01). Pretreatment with melatonin significantly increased the activity of GSH‐Px (20.35±5.27 to 28.93±11.01 mU/min mg−1 protein, P<0.05) in fetal rat brain mitochondria, but the activity of SOD did not change significantly. Results indicate that the administration of melatonin to the pregnant rat may prevent the free radical‐induced oxidative mitochondrial damage to fetal rat brain by a direct antioxidant effect and the activation of GSH‐Px.

Different Effects of Oral Conjugated Equine Estrogen and Transdermal Estrogen Replacement Therapy on Size and Oxidative Susceptibility of Low-Density Lipoprotein Particles in Postmenopausal Women

Background—Postmenopausal estrogen replacement therapy (ERT) has an antioxidant effect that opposes the oxidation of LDL particles. Oral ERT-induced increases in plasma triglyceride, however, decrease LDL particle size, which may counteract this antioxidant effect. Because transdermal ERT decreases plasma triglyceride, it may not decrease LDL particle size and may preserve estrogen’s antioxidant effect. The present study investigates whether transdermal ERT can eliminate the adverse effects of oral ERT on the size and oxidative susceptibility of LDL in postmenopausal women. Methods and Results—Postmenopausal women received no treatment (n=12) or were treated with either 0.625 mg oral conjugated equine estrogen daily (n=16) or with transdermal estradiol (50 &mgr;g/d, n=16) for 3 months. Plasma lipids and the diameter of LDL particles were determined. Susceptibility of LDL to oxidation was analyzed by incubation with CuSO4 and subsequent measurement of thiobarbituric acid reactive substance (TBARS) concentrations. Oral ERT significantly increased plasma triglyceride and decreased LDL diameter but did not affect LDL-derived TBARS concentrations. In contrast, transdermal ERT significantly decreased the concentrations of plasma triglyceride and LDL-derived TBARS and significantly increased LDL diameter. Estrogen-induced changes in LDL diameter correlated negatively with changes in plasma triglyceride (r =−0.51, P <0.001) and LDL-derived TBARS (r =−0.50, P <0.001). Conclusions—Because transdermal, but not oral ERT, decreases plasma triglyceride and produces larger LDL particles that are resistant to oxidation, the antioxidant effect of estrogen can be preserved.

Increased killer inhibitory receptor KIR2DL1 expression among natural killer cells in women with pelvic endometriosis.

OBJECTIVE To investigate the host immunologic response to endometriosis in terms of killer inhibitory receptor (KIR) expression by natural killer (NK) cells. DESIGN Case-control study of immunologic markers. SETTING University hospital. PATIENT(S) We compared cells from Japanese women with laparoscopically diagnosed endometriosis to cells from 40 women with other laparoscopic diagnoses. INTERVENTION(S) Peripheral venous blood sampling and laparoscopic peritoneal fluid collection. MAIN OUTCOME MEASURE(S) Flow cytometry was used to assess expression of KIR by NK cells in the cell samples. RESULT(S) The percentage of cells that expressed KIR2DL1 among NK (KIR2DL1(+)NK) cells in peritoneal fluid and peripheral blood was significantly higher in women with endometriosis than in controls. The proportion of KIR2DL1(+)NK cells in peripheral blood NK cells before and 1 month after laparoscopic surgery did not differ significantly. CONCLUSION(S) The proportion of KIR2DL1(+)NK cells was increased in peritoneal fluid and peripheral blood in women with endometriosis; this difference is probably related to NK cell suppression in endometriosis. This increase in KIR2DL1 expression by NK cells may represent a risk factor in the pathogenesis of endometriosis.

Melatonin stimulates glutathione peroxidase activity in human chorion

In preeclampsia, placental production of lipid peroxides is abnormally increased, while placental glutathione peroxidase (GSH‐Px) and superoxide dismutase (SOD) activities are decreased. Administration of melatonin, a powerful scavenger of oxygen free radicals, also may protect the placenta from free radical‐induced damage by increasing the activity of antioxidant enzymes. To test this hypothesis we administered melatonin to pregnant women before they underwent voluntary interruption of pregnancy between 7 and 9 wk of gestation. Melatonin (6 mg) was administered orally at 12:00 hr, and samples of chorion and maternal blood were obtained at the time of the procedure, 1, 2 or 3 hr later. We measured the melatonin concentration in maternal serum and activities of GSH‐Px and SOD and levels of melatonin in chorionic homogenates. Melatonin administration was reflected by markedly increased melatonin concentrations in maternal serum and in chorion, with peak levels achieved 1 hr after melatonin administration (serum, 46.87±10.87 nM/L; chorionic homogenate, 4.36±1.56 pmol/mg protein). Between 1 and 3 hr after melatonin administration, GSH‐Px activity in chorionic homogenates increased significantly (P<0.001), with peak levels occurring at 3 hr (51.68±3.22 mU/mg protein per min, 137.3% of GSH‐Px activity in untreated control subjects). No significant changes in chorionic SOD activity occurred during the 3‐hr post‐administration period. These results indicate that exogenous melatonin increases GSH‐Px activity in the chorion and thereby may protect indirectly against free radical injury. Melatonin could be useful in treating preeclampsia and possibly other clinical states involving excessive free radical production, such as intrauterine fetal growth retardation and fetal hypoxia.

Comparison of four malignancy risk indices in the preoperative evaluation of patients with pelvic masses

OBJECTIVE The aim of this study was to evaluate the ability of four malignancy risk indices (RMI 1, RMI 2, RMI 3, and RMI 4), incorporating menopausal status, serum CA125 levels, and ultrasound findings, to discriminate a benign from a malignant pelvic mass. STUDY DESIGN This is a retrospective study of 253 women admitted to the Department of Obstetrics and Gynecology of Kochi Medical School, between January 2002 and April 2005 for surgical exploration of pelvic masses. To diagnose ovarian cancer, the sensitivity, specificity, and positive predictive value of serum CA125, ultrasound findings and menopausal status were taken separately and combined into RMI 1, RMI 2, RMI 3, and RMI 4. RESULTS This study confirms that, for the diagnosis of malignancy, four malignancy risk indices were more accurate than menopausal status, serum CA125 levels, and ultrasound findings separately. The accuracy of the RMI 4 was better than RMI 1 (P=0.0013), RMI 2 (P=0.0009) and RMI 3 (P=0.0013). The RMI 4 at a cutoff level of 450 yielded a sensitivity of 86.8%, a specificity of 91.0%, a positive predictive value of 63.5%, a negative predictive value of 97.5%, and an accuracy of 90.4%. CONCLUSION We found that, in the discrimination between benign and malignant pelvic disease, the RMI 4 method was more reliable than RMI 1, RMI 2 and RMI 3. The RMI 4 method is a simple technique that can be used in gynecology clinics as well as less-specialized centers.

Maternally administered melatonin protects against ischemia and reperfusion-induced oxidative mitochondrial damage in premature fetal rat brain

Abstract:  We investigated the oxidative susceptibility of the brain and the effect of maternally administered melatonin on ischemia/reperfusion‐induced cerebral damage in premature fetal rat. Fetal brain mitochondria was separated on the 16th and 19th days of pregnant rats and the respiratory control index (RCI) was measured as an indicator of mitochondrial respiratory activity in the presence or absence of xanthine and xanthine oxidase. The utero–ovarian arteries were occluded bilaterally for 20 min in female rats on day 16 of pregnancy to induce fetal ischemia. Reperfusion was achieved by releasing the occlusion and restoring circulation for 30 min. A sham operation was performed in control rats. Melatonin (10 mg/kg) or vehicle was injected intraperitoneally into the dams 60 min prior to occlusion. The RCI and concentration of thiobarbituric acid‐reactive substances (TBARS) in fetal brain mitochondria were measured. The addition of xanthine and xanthine oxidase significantly decreased mitochondrial RCI at both the 16‐ and 19‐day‐old fetal brain. Xanthine and xanthine oxidase‐induced reduction in RCI was significantly greater in the 16‐day‐old fetal brain than that in the fetal brain from the 19th day of pregnancy. Ischemia/reperfusion significantly reduced RCI and elevated TBARS concentrations in the 16‐day‐old fetal brain mitochondria. Melatonin treatment reversed ischemia/reperfusion‐induced reduction in RCI (2.22 ± 0.10 to 2.53 ± 0.08, P < 0.01) and elevation in TBARS concentrations (13.50 ± 1.82 nmol/mg protein to 8.80 ± 0.78 nmol/mg protein, P < 0.01), resulting in values similar to those in untreated, sham‐treated animals. Results indicate that brain mitochondria in the premature fetal rats appear to be more susceptible to oxidative damage. Melatonin administration to pregnant rats may prevent ischemia/reperfusion‐induced oxidative mitochondrial damage in premature fetal brain.

Intercellular Adhesion Molecule-1 and Hepatocyte Growth Factor in Human Endometriosis: Original Investigation and a Review of Literature

Defects in the cell-mediated immune system may play a role in the pathogenesis or progression of pelvic endometriosis. Possible mediators include macrophages, interleukins-1 and -6, and tumor necrosis factor-α. More recent work points to the involvement of adhesion molecules and growth factors. To clarify the pathogenesis of endometriosis, we compared the characteristics of soluble intercellular adhesion molecule-1 (soluble ICAM-1) and hepatocyte growth factor (HGF) in women with and without endometriosis. We found that, in patients with endometriosis, the concentrations of soluble ICAM-1 in peritoneal fluid increased and interfered with the activity of natural killer cells. We also found that HGF secretion was significantly increased in cultured endometrial stromal cells, and that HGF stimulated the proliferation and migration of, and morphogenic changes in, endometrial epithelial cells. HGF and ICAM-1 play important roles in the initiation and regulation of endometriotic lesions on the microenvironment level. The increased secretion of HGF by eutopic endometrial stromal cells may contribute to the pathogenesis of endometriosis, whereas the increased levels of soluble ICAM-1 may impair natural killer cell activity and accelerate the progression of the disease.

Increased secretion of hepatocyte growth factor by eutopic endometrial stromal cells in women with endometriosis

OBJECTIVE We evaluated the secretion of hepatocyte growth factor by eutopic endometrial stromal cells in vitro. DESIGN Eutopic endometrial stromal cells were isolated and the culture supernatants were collected after 48 hours of incubation. The secretion of hepatocyte growth factor was analyzed by ELISA. SETTING Department of Obstetrics and Gynecology of Tohoku University Hospital. PATIENT(S) Specimens of endometrium in the proliferative phase were excised from 11 patients who were undergoing laparoscopy for infertility. Six of the patients had endometriosis and five did not. None of the patients had received hormonal treatment before surgery. RESULT(S) Hepatocyte growth factor secretion was found to be increased significantly in cultured endometrial stromal cells from the infertile patients with endometriosis compared with those without endometriosis. CONCLUSION(S) The secretion of hepatocyte growth factor was up-regulated in eutopic endometrial stromal cells from patients with endometriosis. An increase in hepatocyte growth factor production may be characteristic of endometriosis and may be involved in the pathophysiology of this disease.