Ikunosuke Tsuneki

Mutational Analysis of BRCA1 Gene in Ovarian and Breast-ovarian Cancer Families in Japan

We analyzed the alteration of BRCAI in DNA obtained from 83 individuals of 13 Japanese site‐specific ovarian cancer families and 6 breast‐ovarian cancer families. Six germline mutations were detected in 7 families, which consisted of 4 breast‐ovarian cancer and 3 site‐specific ovarian cancer families, by single‐strand conformation polymorphism analysis, followed by direct sequence determination. The mutations included three framcshifts, two nonsense mutations, and one missense mutation causing loss of a zinc‐binding motif. The frequency of loss of heterozygosity at the microsatellite markers on the BRCAI gene was 57% (8 of 14 cases) in site‐specific ovarian cancer families, and 100% (6 of 6 cases) in breast‐ovarian cancer families. All tumors of the patients carrying a mutation of BRCAI showed deletion of wild‐type alleles, implicating BRCAI as a tumor suppressor gene. Tbese results suggest tbat germline mutations of the BRCAI gene play an important role in the carcinogen‐esis of breast and/or ovarian cancer in a majority of breast‐ovarian cancer families and in some site‐specific ovarian cancer families.

Primary strumal carcinoid tumor of the ovary with multiple bone and breast metastases

Although primary carcinoid tumor of the ovary is an extremely rare neoplasm, survival is excellent if the disease is confined to one ovary. Herein, we present a case of primary strumal carcinoid tumor of the ovary, stage IA, borderline malignancy, in a 34‐year‐old woman. Histological findings of the right ovary indicated higher atypical nuclei, higher mitotic rate and focal necrosis of tumorous cells in some areas, findings that are compatible with atypical carcinoid of the lung. Immunohistochemical staining was positive for synaptophysin, neuron‐specific enolase, chromogranin A, Ki‐67, topoisomerase IIα, peptide YY, and thyroglobulin. Three and a half years postoperatively, multiple bone and breast metastases were found and anticancer chemotherapy was ineffective. The results in the present case indicate that an ovarian carcinoid tumor found to be ‘atypical carcinoid’ according to pulmonary carcinoid criteria or immunohistochemical staining (i.e. highly positive for topoisomerase IIα and Ki‐67) may have a poor prognosis.

Analysis of TH1 and TH2 cells by intracellular cytokine detection with flow cytometry in patients with ovarian cancer

Objective: Our aim is to assess the immune status of patients with ovarian cancer by analyzing the ratio of T helper type 1 (TH1) to T helper type 2 (TH2) populations in peripheral blood lymphocytes (PBL). Methods: We examined TH1/TH2 ratios in PBL obtained from 21 ovarian cancer patients who had just received postoperative chemotherapy, by detecting the intracellular IFN-γ and IL-4 production with 3-color flow cytometry. Additionally, we evaluated the influence of a granulocyte-colony stimulating factor (G-CSF) injection on TH1 and TH2 populations for a rescue of granulocytopenia due to the chemotherapy. Results: We could not find any significant difference of the TH1/TH2 ratios in terms of age, International Federation of Gynecology and Obstetrics (FIGO) clinical stage and clinical tumor status. As for the clinical tumor status, however, the patients with residual cancer had a higher TH1/TH2 ratio, though it was not statistically significant (p = 0.15). Anticancer chemotherapy is also considered to lead to the immunosuppressive state of the patients. TH1 and TH2 populations of PBL in the patients during chemotherapy showed an unfavorable imbalance that was shifted from TH1 to TH2 10 days after anticancer drug administration (p = 0.049). G-CSF administration, on the other hand, was likely to induce a cell population shift from TH2 to TH1 assessed by the intracellular cytokine assay (p = 0.051), and never induced an unfavorable imbalance from TH1 to TH2 in the T cell population by a 1-day injection of G-CSF. Conclusion: Together, these data indicate that the TH1/TH2 ratio analyzed by intracellular cytokine flow cytometry seems to be a good indicator to assess the immune status in cancer.

Examination of the taste disorder associated with gynecological cancer chemotherapy

OBJECTIVE Taste disturbance is known to occur as one of the adverse events associated with chemotherapy for gynecological cancer, but few studies have attempted to assess it in practical terms. Therefore, a range of taste tests was performed in gynecological cancer patients. METHODS The patients were 23 women with gynecological cancer being treated with anticancer agents. Subjective symptoms of altered taste were classified, and objective findings were obtained with the following four gustatory tests: serum trace element (zinc, copper, iron) levels, tongue cultures, electrogustometry, and the filter paper disc tests. RESULTS Of the 23 subjects, 11 perceived taste disturbances. The serum zinc level was consistently below the lower limit of normal. On tongue cultures, indigenous bacteria were seen in all patients during the entire treatment period. Electrogustometry revealed a tendency for the development of hypogeusia in the chorda tympani nerve field. Conversely, hypergeusia tended to develop gradually in the greater petrosal nerve field. The filter paper disc test revealed a tendency for the development of hypergeusia for sweetness, saltiness, and sourness in the chorda tympani nerve field. Hypogeusia for bitterness tended to develop with increasing number of chemotherapy cycles. The glossopharyngeal nerve field exhibited the same tendencies as observed in the chorda tympani nerve field. In the greater petrosal nerve field, there was a tendency for the development of hypergeusia for sweetness, sourness, and bitterness. CONCLUSIONS Abnormal test results were seen in half of patients after cancer chemotherapy.

Effectiveness of the cyclic administration of dienogest in a case of pathological disappearance of intestinal endometriosis.

We have reported good control of atypical genital bleeding when using a cyclic administration of dienogest (repeated 4-week cycles, each consisting of the administration of 2 mg/day of dienogest for 3 weeks, followed by 1 week of drug withdrawal) in patients with endometriosis. Herein, we report the effectiveness of the long-term cyclic administration (22 months) of dienogest in a case of pathological disappearance of intestinal endometriosis diagnosed by endoscopy and histology of the lower gastrointestinal tract. There is no recurrent sign after 16 months of the treatment being stopped. Atypical genital bleeding during treatment was 3–5 days a month in each cycle. Compliance was good, so we could continue the therapy. The long-term cyclic administration of dienogest in patients with intestinal endometriosis may have significant merit.

Outcomes of treatment with cyclic administration of dienogest after ovarian endometriotic cystectomy

Abstract To evaluate the outcomes of patients treated with cyclic administration of dienogest after ovarian endometriotic cystectomy, following the completion of treatment. We retrospectively evaluated 26 patients treated with dienogest (2 mg/day) after cystectomy (revised American Society for Reproductive Medicine [r-ASRM] stage III–IV) in a pilot study. Dienogest was administered cyclically, for a total of six cycles, each comprising three weeks on and one week off. Outcomes of interest included severity of menstrual pain and recurrence of cysts at baseline, during the immediate post-treatment period and at the final outpatient follow-up. The mean outpatient follow-up period was 45.0 months. The visual analog scale score for menstrual pain following 6 cycles of dienogest treatment was significantly lower than that at baseline; it remained low at the final follow-up. The recurrence rates of cysts were 4% and 21% at 24 and 48 months after the completion of dienogest treatment, respectively. Six patients with recurrent disease were all classified as having r-ASRM stage IV. Our results suggest that cyclic dienogest for six months after cystectomy could relieve menstrual pain and reduce the recurrence of cysts, for approximately four years. The necessary treatment period for patients with r-ASRM stage IV disease requires further study. Chinese abstract 研究的目的为评估地诺孕素周期性治疗对卵巢子宫内膜异位囊肿切除术后患者的治疗效果。我们回顾性地分析了一项试验研究中26位内异囊肿切除术（修订版美国生殖医学会[r-ASRM] Ⅲ-Ⅳ期）后服用地诺孕素（2mg/天）的患者。地诺孕素行周期式给药方式，共6个周期，每周期服药3周停药1周。研究观察基线期、治疗结束后以及末次随访时患者的痛经程度和囊肿复发情况。平均随访期为45个月。6个周期治疗结束时患者的痛经视觉模拟评分显著低于基线期；末次随访时仍保持较低水平。异位囊肿的复发率在地诺孕素治疗结束后的24个月与48个月分别为4%和21%。研究中6位复发的患者均为r-ASRM Ⅳ期。研究结果提示，在囊肿切除术后行6个月的地诺孕素周期性治疗可以有效改善痛经症状，降低囊肿的复发率，且效果可能维持4年。在将来的研究中还需要进一步研究r-ASRM Ⅳ期患者的治疗情况。

Ovarian tumor cell detection in peripheral blood progenitor cells harvests by RT-PCR

Background.  To evaluate the frequency of tumor cell contamination in autologous peripheral‐blood progenitor cells from patients with ovarian cancer, and to determine the impact of infusing such cells on relapses after high‐dose chemotherapy.

Circulating CD34+ cells to predict the adequate harvest of peripheral blood progenitor cells in platinum-based chemotherapy.

Abstract  Objective: The aim of this study was to clarify the appropriate timing for peripheral blood progenitor cells (PBPC) harvesting after platinum-based mobilization chemotherapy by measurement of the circulating CD34+ cell concentrations. Patients and Methods: PBPC were collected for autotransplantation via a total of 68 leukaphereses in 16 patients with gynecological cancer. Circulating CD34+ cell concentrations were measured by CD34-side scatter parameter analysis. Results: Data could be fitted into a linear regression line described by the equation y=0.33+4.14×x (R2=0.256), where y=the number of harvested colony-forming unit granulocyte macrophage (CFU-GM (×105/kg) and x=the percentage of circulating CD34+ cells and y=1.747+44.53×x (R2= 0.475), where y=the number of harvested CD34+ cells (×106/kg) and x=the percentage of circulating CD34+ cells. Failure to mobilize sufficient CFU-GM numbers (>1×105/kg) occurred in 29 of 31 leukaphereses when the percentage of circulating CD34+ cells was less than 0.10%. Conclusions: Harvesting procedure may be avoided when circulating CD34+ cells showed less than 0.10% after platinum-based mobilization chemotherapy.

Relief of uterine bleeding by cyclic administration of dienogest for endometriosis

Abstract Objective: This study assessed the relief of uterine bleeding and clinical symptoms during cyclic administration of dienogest for the treatment of endometriosis. Methods: In total, 25 patients undergoing ovarian cyst enucleation and given dienogest participated in this study. Dienogest 2 mg/day was administered for 3 weeks, and the drug was then withdrawn for 1 week (cyclic administration of dienogest). This 4-week cycle was repeated six times. Patients’ records were prospectively analyzed for the number of days on which any uterine bleeding occurred, as well as menstrual pain before and after the start of dienogest administration were evaluated with a view to using the data obtained herein as the basis. Results: During the period of cyclic administration of dienogest, uterine bleeding occurred on 5.8 to 7.7 days per 4-week period on an average through cycles. Of uterine bleeding episodes, menstruation-like uterine bleeding was present in about 80% of patients. The visual analog scale (VAS) value for menstrual pain significantly decreased from 3.8 before dienogest administration after surgery to 1.5 at the completion of cycle 1, VAS remained low thereafter. Conclusion: These results raise the possibility that cyclic administration of dienogest may relieve lessen uterine bleeding, a major adverse event and menstrual pain. Chinese abstract 目的：本研究评估周期应用地诺孕素治疗子宫内膜异位症时子宫出血和临床症状缓解的情况。 方法：共有25例进行了卵巢囊肿摘除术并给予地诺孕素的患者参与了此项研究。地诺孕素2 mg /天，给药3周，然后停药1周（周期应用地诺孕素）。这样4周一个周期，重复六次。记录患者的数据进行前瞻性的分析：包括对任一发生子宫出血的天数，评估用地诺孕素前后的月经疼痛情况。结果：周期应用地诺孕素期间，以每4周的时间作为平均周期，子宫出血发生为5.8到7.7天。子宫出血的发生，像月经的子宫出血发生在大约80%的患者。痛经的视觉模拟量表（VAS）值从术后用地诺孕素前的3.8明显的减少为第一周期结束后的1.5，此后VAS仍然很低。结论：这些结果表明，周期性应用地诺孕素可能可以缓少子宫出血，这是一个主要的不良事件并且减轻痛经。

Circulating highly fluorescent reticulocytes to predict the adequate harvesting of peripheral blood progenitor cells in platinum-based chemotherapy.

We prospectively evaluated whether peripheral blood progenitor cells (PBPC) yield from a single leukapheresis could be predicted by measurement of circulating highly fluorescent reticulocytes (HFR). PBPC were collected from 46 leukaphereses in 15 patients with gynecological cancer following platinum-based chemotherapy. Once a level of at least 5.0% HFR was achieved, sufficient PBPC were collected in a single harvest in 71% of the procedures. Whereas, failure to mobilize sufficient PBPC occurred in 24 of 28 leukaphereses when the percentage of circulating HFR was less than 5.0%. In conclusion, circulating HFR may aid in the efficiency of PBPC collections in platinum-based chemotherapy.