Hitoshi Morioka

Oxidative stress impairs oocyte quality and melatonin protects oocytes from free radical damage and improves fertilization rate

Abstract:  We investigated the relationship between oxidative stress and poor oocyte quality and whether the antioxidant melatonin improves oocyte quality. Follicular fluid was sampled at oocyte retrieval during in vitro fertilization and embryo transfer (IVF‐ET). Intrafollicular concentrations of 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG) in women with high rates of degenerate oocytes were significantly higher than those with low rates of degenerate oocytes. As there was a negative correlation between intrafollicular concentrations of 8‐OHdG and melatonin, 18 patients undergoing IVF‐ET were given melatonin (3 mg/day), vitamin E (600 mg/day) or both melatonin and vitamin E. Intrafollicular concentrations of 8‐OHdG and hexanoyl‐lysine adduct were significantly reduced by these antioxidant treatments. One hundred and fifteen patients who failed to become pregnant with a low fertilization rate (50%) in the previous IVF‐ET cycle were divided into two groups during the next IVF‐ET procedure; 56 patients with melatonin treatment (3 mg/day) and 59 patients without melatonin treatment. The fertilization rate was improved by melatonin treatment compared to the previous IVF‐ET cycle. However, the fertilization rate was not significantly changed without melatonin treatment. Oocytes recovered from preovulatory follicles in mice were incubated with H2O2 for 12 hr. The percentage of mature oocytes with a first polar body was significantly reduced by addition of H2O2 (300 μm). The inhibitory effect of H2O2 was significantly blocked by simultaneous addition of melatonin. In conclusion, oxidative stress causes toxic effects on oocyte maturation and melatonin protects oocytes from oxidative stress. Melatonin is likely to improve oocyte quality and fertilization rates.

Tumor-antigen TA-4 in the detection of recurrence in cervical squamous cell carcinoma

A radioimmunoassay for a tumor‐antigen (TA‐4) of squamous cell carcinoma was used to detect the recurrence in patients with squamous cell carcinoma of the uterine cervix. Of 17 patients who had recurrence, 15 cases showed reappearance of serum TA‐4 levels. Reappearance of serum TA‐4 was faster than other clinical signs of recurrence in 11 cases. These results indicated that TA‐4 assay would be a useful aid for the detection of recurrence in cervical squamous cell carcinoma.

Value of tumor-antigen (TA-4) of squamous cell carcinoma in predicting the extent of cervical cancer

The value of a tumor‐antigen (TA‐4) of squamous cell carcinoma in evaluating the extent of disease was studied in patients who primarily had surgical treatments with a diagnosis of cervical squamous cell carcinoma. Pretreatment serum TA‐4 levels were determined by a radioimmunoassay method. The extent of disease was determined by surgical evaluation and postoperative histologic examinations. Furthermore, all patients were followed for at least two years, and those patients who developed recurrence at the sites which were unresectable by radical hysterectomy were retrospectively regarded as the unresectable case. It was found that serum TA‐4 levels reflected the extent of disease, and that high pretreatment TA‐4 levels indicated the presence of the widespread tumor which was unresectable by radical hysterectomy.

Tumor antigen of human cervical squamous cell carcinoma. Correlation of circulating levels with disease progress

A double‐antibody radioimmunoassay method was used for serial determinations of a tumor‐antigen (TA‐4) of cervical squamous cell carcinoma, and the correlation of serum antigen levels with the disease progress was investigated in 23 patients with cervical squamous cell carcinoma. Ten cases with widespread metastases received radiotherapy and/or chemotherapy. Nine of these cases who showed progression of the disease had a corresponding increase in serum antigen levels, while one case who had regression of the disease showed a corresponding decrease in serum antigen levels. Thirteen patients received radical surgery, and in all of these, high pretreatment antigen levels declined to undetectable levels 1 or 2 weeks after surgery. A panel of coded sera from the NCI‐Mayo Clinic Serum Bank was also studied for evaluating the specificity of the assay. Thirteen of 25 patients (52%) with cervical squamous cell carcinoma showed positive serum antigen levels, while only one of 58 control cases (1.7%) showed false‐positive result. These results suggest that serial TA‐4 determinations may provide a useful method for evaluating regression or progression of the disease. Cancer 43:585–590, 1979.

Prognostic significance of the tumor antigen TA-4 in squamous cell carcinoma of the uterine cervix

Prognostic values of a tumor antigen (TA-4) of squamous cell carcinoma were studied in 135 patients with invasive squamous cell carcinoma of the uterine cervix. In order to evaluate the host defense responses against cancer, the percentage of lymphocytes (percentages of lymphocytes in total leukocyte counts in the peripheral blood) was also determined simultaneously in each case. All patients were followed up for 2 years. Sixty patients with Stage II disease underwent a radical operative procedure, and all other cases (75 cases) were treated primarily with radiation therapy. In both groups of patients, the survival rate or disease-free rate was significantly worse in those with TA-4 levels of greater than or equal to 15 microunits/ml compared to that of those with lower levels. The prognosis was particularly poor in those patients who had TA-4 levels of greater than or equal to 15 microunits/ml and lymphocyte percentages of less than 30. It was concluded that the simultaneous determinations of serum TA-4 and lymphocyte percentages would be useful in predicting the prognosis of cervical squamous cell carcinoma.

Genetic aberrations detected by comparative genomic hybridization predict outcome in patients with endometrioid carcinoma.

Endometrial cancer progression is determined by a complex pattern of multiple genetic aberrations, but how these aberrations affect prognosis is unknown. In this study, we undertook a genome‐wide screening to detect genetic changes by comparative genomic hybridization (CGH) in 51 tumors from patients with primary endometrioid carcinoma of the uterine corpus. The observed genetic changes were subsequently correlated with the progression of the disease and the clinical outcome in each case. The average number of genetic aberrations (copy number gains and losses) was significantly greater in non‐surviving patients than in disease‐free patients (12.6 vs. 2.7, P < 0.0001). According to multivariate analysis, lymph node metastasis (P = 0.015), cervical involvement (P = 0.007) and one or more copy number losses at 9q32–q34, 11q23, or Xq12–q24 (P = 0.023) were significantly predictive of death from the disease. Interestingly, lymph node metastasis was significantly associated with copy number gains at 8q22–q23 and 8q24–qter (P = 0.003 and P = 0.025, respectively). Moreover, cervical involvement was also correlated significantly not only with gains of 8q22–q23 and 8q24–qter but also with loss of 11q23 (P = 0.04, 0.0003, and P = 0.009, respectively). These results suggest that analysis of genetic changes may help predict clinical outcome and the presence of metastatic disease as well as assist in therapeutic decision making for patients with endometrioid carcinoma. Genes Chromosomes Cancer 29:75–82, 2000.

Luteal blood flow and luteal function

BackgroundBlood flow in the corpus luteum (CL) is associated with luteal function. The present study was undertaken to investigate whether luteal function can be improved by increasing CL blood flow in women with luteal phase defect (LFD).MethodsBlood flow impedance in the CL was measured by transvaginal color-pulsed-Doppler-ultrasonography and was expressed as a resistance index (RI). The patients with both LFD [serum progesterone (P) concentrations < 10 ng/ml during mid-luteal phase] and high CL-RI (≥ 0.51) were given vitamin-E (600 mg/day, n = 18), L-arginine (6 g/day, n = 14) as a potential nitric oxide donor, melatonin (3 mg/day, n = 13) as an antioxidant, or HCG (2,000 IU/day, n = 10) during the subsequent menstrual cycle.ResultsIn the control group (n = 11), who received no medication to increase CL blood flow, only one patient (9%) improved in CL-RI and 2 patients (18%) improved in serum P. Vitamin-E improved CL-RI in 15 patients (83%) and improved serum P in 12 patients (67%). L-arginine improved CL-RI in all the patients (100%) and improved serum P in 10 patients (71%). HCG improved CL-RI in all the patients (100%) and improved serum P in 9 patients (90%). Melatonin had no significant effect.ConclusionVitamin-E or L-arginine treatment improved luteal function by decreasing CL blood flow impedance. CL blood flow is a critical factor for luteal function.

Melatonin as a new drug for improving oocyte quality

BackgroundAlthough recent technical advances have benefited infertile couples, inadequate embryo development as a result of poor quality oocytes still contributes to infertility. The purpose of the present study was to evaluate melatonin as a drug for improving oocyte quality in such cases.MethodsTwenty-seven women from whom fewer than three fertilized embryos were grown and who failed to fall pregnant in previous treatment cycles were enrolled in the current prospective clinical study. Subjects took 1 mg or 3 mg tablets of melatonin orally at 22:00 h from the fifth day of the previous menstrual cycle to the day they were injected with human chorionic gonadotropin. The numbers of mature follicles, retrieved oocytes, degenerate oocytes, and fertilized embryos were compared to their previous data without melatonin (the control cycle).ResultsIntrafollicular melatonin concentrations were significantly increased, and intrafollicular lipid peroxide concentrations showed a tendency towards lower levels in the 3 mg melatonin treatment cycles compared with the control cycles. The number of degenerate oocytes was significantly reduced, and the number of fertilized embryos showed a tendency towards an increase in the 3 mg cycle compared to the control cycle. Three women succeeded in falling pregnant.ConclusionMelatonin is likely to become the drug of choice for improving oocyte quality in women who cannot fall pregnant because of poor quality oocytes.

Individualization of the Cutoff Value for Serum Squamous-Cell Carcinoma Antigen Using a Sensitive Enzyme Immunoassay

A sensitive enzyme immunoassay (IMx) for the squamous-cell carcinoma (SCC) antigen was used to evaluate low concentrations of this marker in the blood circulation. The assay can detect 0.05 ng/ml of serum SCC antigen, with intra- and interassay variabilities of 4.8 and 8.9%, respectively. Daily changes in serum SCC antigen were determined in healthy volunteers and in patients with cervical squamous-cell carcinoma after complete resection of the primary tumor, which revealed that, although the absolute values were variable from one case to another, the daily changes were relatively constant in each case, with a mean daily variation of 24.1 +/- 3.7% (mean +/- SE). The determination of the cutoff value for each individual patient may be effective for the early detection of an abnormal rise of circulating tumor marker, and will be useful in detecting small tumor foci during follow-up after treatment.

Luteal blood flow in patients undergoing GnRH agonist long protocol.

BackgroundBlood flow in the corpus luteum (CL) is closely related to luteal function. It is unclear how luteal blood flow is regulated. Standardized ovarian-stimulation protocol with a gonadotropin-releasing hormone agonist (GnRHa long protocol) causes luteal phase defect because it drastically suppresses serum LH levels. Examining luteal blood flow in the patient undergoing GnRHa long protocol may be useful to know whether luteal blood flow is regulated by LH.MethodsTwenty-four infertile women undergoing GnRHa long protocol were divided into 3 groups dependent on luteal supports; 9 women were given ethinylestradiol plus norgestrel (Planovar) orally throughout the luteal phase (control group); 8 women were given HCG 2,000 IU on days 2 and 4 day after ovulation induction in addition to Planovar (HCG group); 7 women were given vitamin E (600 mg/day) orally throughout the luteal phase in addition to Planovar (vitamin E group). Blood flow impedance was measured in each CL during the mid-luteal phase by transvaginal color-pulsed-Doppler-ultrasonography and was expressed as a CL-resistance index (CL-RI).ResultsSerum LH levels were remarkably suppressed in all the groups. CL-RI in the control group was more than the cutoff value (0.51), and only 2 out of 9 women had CL-RI values < 0.51. Treatments with HCG or vitamin E significantly improved the CL-RI to less than 0.51. Seven of the 8 women in the HCG group and all of the women in the vitamin E group had CL-RI < 0.51.ConclusionPatients undergoing GnRHa long protocol had high luteal blood flow impedance with very low serum LH levels. HCG administration improved luteal blood flow impedance. This suggests that luteal blood flow is regulated by LH.