OCT4 expression in human uterine myometrial stem/progenitor cells.

BACKGROUND: The transcription factor, octamer-binding transcription factor 4 (OCT4)/POU5F1, is expressed in embryonic stem cells, germ cells and some types of adult stem cells. Human OCT4 encodes two isoforms, OCT4A and OCT4B. While OCT4A plays a crucial role in the maintenance of stem cell properties, including pluripotency, whereas OCT4B does not. We previously reported that human myometrium contains side population cells (myoSP) with a Hoechst 33 342 low-fluorescent profile. These cells exhibit phenotypic and functional characteristics of myometrial stem cells. The objective of this study was to investigate the comparative expression of OCT4 in the stem/progenitor cell population of the human myometrium. METHODS: Human myometrial tissue samples were collected from 18 consenting patients who underwent hysterectomy because of benign gynecological diseases. The resultant isolated or cultured myometrial cells and isolated myoSP were subjected to semi-quantitative and real-time RT-PCR analyses, immunoblot analyses and immunohistochemistry. RESULTS: RT-PCR and immunoblot analyses revealed that OCT4 mRNA and OCT4 protein were detectable in some (but not all) myometrial samples. Immunohistochemistry showed that OCT4 protein was confined to the nuclei of relatively few cells in myometrial tissues expressing OCT4 mRNA. OCT4 and OCT4A transcripts, but not those of OCT4B, were more abundant in myoSP than in non-myoSP, as determined by real-time and semi-quantitative RT-PCR analyses. CONCLUSIONS: Relatively few myometrial cells express OCT4 protein. OCT4 mRNA, in particular OCT4A mRNA, is up-regulated in myoSP that have been reported to exhibit stem cell-like properties. Taken together, the present results indicate that the myoSP population is enriched in OCT4 mRNA.

Analysis of serum FSH bioactivity in a patient with an FSH-secreting pituitary microadenoma and multicystic ovaries: A case report.

FSH-secreting pituitary adenoma (FSHoma) is often associated with increased levels of serum FSH and ovarian hyperstimulation syndrome (OHSS). The OHSS has historically been attributed to elevated FSH production by the FSHoma; however, some FSHoma patients with OHSS have normal serum FSH levels. OHSS may result not from increased FSH levels, but also from increased bioactivity of the FSH derived from the adenoma. To address this, we measured the FSH bioactivity in the serum of a 40-year-old woman with an FSHoma and OHSS, whose FSH levels were normal. Chinese hamster ovary cells stably expressing FSH receptors were prepared and transfected with a cAMP-responsive element-driven luciferase reporter plasmid. Cells were then treated with recombinant human FSH (rhFSH), the patient's sera, or sera from controls, collected at different time points, and subjected to a luciferase assay. Luciferase activity was increased in response to rhFSH in a dose-dependent manner. The responsiveness was further augmented by co-addition of a 3-methyl isobutylxanthine, which improved the sensitivity of our assay. Unexpectedly, the serum FSH bioactivity/immunoactivity ratio of the patient was mostly equal to that of normal subjects. This was confirmed with a granulosa cell aromatase assay. This case report suggests that alternate explanations may exist for the OHSS phenotype seen in some FSHoma patients.

Synergism of diethylstilbestrol and other carcinogens in concurrent development of hepatic, mammary, and pituitary tumors in castrated male rats.

Castrated male WF rats, given implants of pellets containing 5.0 mg diethylstilbestrol (DES), were given N-butyl-N-nitrosourea (NBU) in small amounts, which alone produced no mammary tumors in intact female rats. Treatment resulted in the high yield of hepatic tumors (HT), mammary tumors (MT), and pituitary tumors (PT) concurrently in each rat. If animals were further treated with prolactin, the development of HT and MT was accelerated, whereas that of PT was suppressed. None of the intact or castrated rats receiving NBU and/or prolactin developed tumors in any tissues if DES treatment was omitted. Exposure of male rats, preconditioned similarly to NBU treatment, to 200 rads of 14.1-MeV fast-neutron radiation also elicited HT, MT, and PT with an efficiency comparable to that of NBU-treated rats. These findings indicate that DES played an essential role in the whole carcinogenic process in each tissue and that castrated male rats, if conditioned properly with estrogens, are useful for the study of the carcinogenesis mechanism in these tissues.

Comparison of 5-fluorouracil with ftorafur in adjuvant chemotherapies with combined inductive and maintenance therapies for gastric cancer.

A group of 243 patients with gastric cancer was subjected to a prospective randomized trial of adjuvant chemotherapy after curative gastrectomy. One of the three arms (regimen A) is induction therapy with a three-drug combination of mitomycin C (MMC), 5-fluorouracil (5-FU), and cytosine arabinoside (ara-C) followed by intermittent oral administration of 5-FU for two years (MFC + F). In regimen B, 5-FU in regimen A is replaced by ftorafur, administered on the same schedule as regimen A (MF'C + F'). Regimen C is the control, in which patients are treated by surgery alone. Regimen A was proven effective for the suppression of relapse in the limited groups of moderately locally advanced cancer; five-year survival rates of the subset of stages I, II, and III were 72.1% with regimen A and 53.1% with regimen C (P less than .05). Regimen B yielded a better survival than the control, although the difference did not reach statistical significance. The results seem to favor 5-FU as adjuvant chemotherapy for gastric cancer.

Expression of epiregulin and amphiregulin in the rat ovary.

We have previously reported that the epidermal growth factor (EGF) family growth factor, epiregulin, is expressed in rat ovarian granulosa cells by induction with pregnant mare serum gonadotropin (PMSG). In this study, we report that amphiregulin, another member of the EGF family, was also induced in the rat ovary by gonadotropin treatment. Northern blot analysis revealed that PMSG treatment induced the expression of both epiregulin and amphiregulin mRNA after 24 h, but the expression then decreased 48 h after treatment. Further treatment with human chorionic gonadotropin (hCG) rapidly induced the expression of both epiregulin and amphiregulin genes and maximal levels were reached 4 h after hCG treatment. A marginal increase in amphiregulin mRNA levels was also observed 6 h after PMSG treatment. In situ hybridization revealed that epiregulin and amphiregulin mRNAs were localized in the granulosa cells of large antral follicles. These spatio-temporal expression patterns were similar to those of cyclo-oxygenase-2 (COX-2) and progesterone receptor (PR). In adult cycling rats, epiregulin and amphiregulin were strongly induced at 1800 and 2000 h on proestrus coinciding with the preovulatory LH surge. An in situ hybridization study also showed that epiregulin and amphiregulin mRNAs were detectable in the granulosa cells of preovulatory ovarian follicles at 2000 h on proestrus, where transcripts of COX-2 and PR were co-localized with those of epiregulin and amphiregulin. These observations suggested that the EGF family members, epiregulin and amphiregulin, may play a role in the ovulatory process of cycling rats as well as in the induction of ovulation in immature rats.

Characteristic echographic features of circumscribed cancer.

Of 964 amputated breast cancers which had been examined preoperatively by ultrasound and mammography, 147 breast (17.0%) contained macroscopically circumscribed tumors and 596 cases (69.0%) showed infiltrative tumors on examination of the cut surface. It is well known that infiltrative types of cancer show ultrasonic images that usually possess clear cut signs of malignancy, such as jagged augmented boundary echoes and acoustic shadows. On the other hand, circumscribed types of cancer do not usually show such signs, but exhibit lobulated shapes and strong, coarse and non-uniform internal echoes. These latter features are intermediate between those of infiltrative cancers and those of solid benign tumors. As such circumscribed cancers are often difficult to diagnose correctly by ultrasonic techniques. The diagnostic accuracy rates by ultrasound were 88.0% in the infiltrative types and 69.4% in the circumscribed types. The ultrasonic pictures of the 147 cases of such circumscribed tumors are classified and discussed.

Febrile convulsions and rolandic discharges.

Clinical and electroencephalographic assessments were performed on 39 children with febrile convulsions showing EEG evidence of rolandic discharges and 23 epileptic children showing rolandic discharges with a preceding history of febrile convulsions. Ten families in which EEGs from the proband and his (or her) sibling or cousin demonstrated rolandic discharges were also described. The results of these studies stress a close genetic relationship between febrile convulsions with rolandic discharges and focal seizures or generalized convulsions that occur during sleep, namely, benign epilepsy of childhood with rolandic discharges. This indicates that the febrile convulsions with rolandic discharges constitute a particular subtype of febrile convulsions.

Relationship between benign epilepsy of children with centro-temporal EEG foci and febrile convulsions.

In order to clarify the relationship between benign epilepsy of children with centro-temporal EEG foci (BECCT) and febrile convulsions (FC), we compared the previous and family histories of FC of one hundred children with BECCT, aged 3 years to 13 years, with those of 100 non-epileptic controls matched for age and sex. The incidences of FC in children with BECCT and non-epileptic controls were 18% and 8%, respectively (P less than 0.05). Forty-eight children (48%) with BECCT and 21 (21%) non-epileptic controls had positive family histories of FC within third-degree relatives (P less than 0.001). Of the fourteen pairs of siblings, one of each having BECCT and the other FC, we conducted an EEG examination in 7 siblings. All of these 7 siblings exhibited rolandic discharges (RD) on EEG. These observations strongly suggested a genetic link between BECCT and FC.

Prognostic and therapeutic values of peritoneal cytology in gastric cancer.

Exfoliative cytology of peritoneal lavage was performed at the time of laparotomy for 458 patients with gastric cancer. Positive cytologic specimens were obtained from 28 per cent of all cases and from 16 per cent of patients with curative surgery. Peritoneal cytology has revealed itself to be a good predictor of prognosis and an indicator for adjuvant therapies. In the group treated with curative surgery, the five year survival rate was 86 per cent for patients with intact serosa and negative cytologic specimens, and 52 per cent for patients with involved serosa and negative cytology. None of the patients who had involved serosa and positive cytologic specimens survived more than five years. A slight benefit of adjuvant chemotherapy was observed in the patients who had curative surgery with negative cytology. Long term immunochemotherapy, including repeated intraperitoneal administration of mitomycin-C, improved the lifespan of patients with non-curative surgery.

Surgical treatment of gastric cancer today.

The utilization of radical operation in gastric cancer cases has led to the development of various operation techniques. With the improvement in perioperative management it can be said that age no longer is a restriction. A study of gastric cancer prognosis indicates an upward trend in survival since 1960 along with the increased detection of early cancer. This suggests the most importance of early detection in increasing the therapeutic results obtained in cases of gastric cancer. In cases with advanced cancer, 5-year survival rate of 44.7 per cent was attained through such aggressive actions as extended dissection of lymph nodes and combined resection of surrounding organs with evaluation of the macroscopic classification of the cancer.

Lithium potassium borate and lithium rubidium borate: new non-linear optical crystals

The title compounds, LiKB(4)O(7) and LiRbB(4)O(7), are newly developed non-linear optical crystals containing two kinds of anionic groups, namely (B(3)O(8))(7-) and (B(5)O(10))(5-). The (B(3)O(8))(7-) groups form infinite spiral chains parallel to the [100] direction, which are interconnected by sharing O atoms with (B(5)O(10))(5-) groups.

[Quantitative evaluation of liver function with 99mTc-GSA scintigraphy using extraction index].

Extraction index (EI5) was introduced to evaluate liver function quantitatively with 99mTc-GSA (GSA) scintigraphy, and it was compared with conventional indices; receptor index (LHL15) and clearance index (HH15). EI5 is expressed as following equation: EI5 = (L5 - L3)/(H3 + H5) * PH/PL where L3, L5: counts at 3 or 5 minutes after the injection in the liver ROI, respectively, H3, H5: counts at 3 or 5 minutes in the heart ROI, respectively; PL, PH: numbers of pixels in the liver- and heart-ROI, respectively. We performed GSA scintigraphy in 40 patients with liver dysfunction and calculated values of the indices. Good correlations were observed between EI5 and liver function tests. Correlation coefficients were almost equal to or higher than those between conventional parameters and liver functional tests. EI5 was thought to be a practical index, and it could be calculated in a short time without aid of computer. Evaluation of local liver function may be possible, because EI5 was corrected with numbers of pixels in the liver- and heart-ROI; which was not considered in the conventional parameters.

[Adjuvant chemotherapy of colorectal cancer--results of prospective randomized trials].

The group of research for colorectal cancer treatments-Kajitani-group (chief T. Kajitani) has carried out the co-operative study for the evaluation of adjuvant chemotherapy after curative resection of colorectal cancer. During the period 1975 and 1978, a series of 1,156 cases of cancer of colon and rectum were entered into the prospective randomized controlled study which consisted of three treatment programs. There included chemotherapy of 2 modes of regimen combining MMC with Tegaful and non adjuvant treatment as control. In colon cancer, adjuvant chemotherapy combining MMC with Tegaful was effective on the increasing of survival rates, especially significantly (p = 0.017) in the cases of Dukes B stage (85-88% vs 69.2% in survival rates of 8 year). In rectal cancer, systemic intravenous administration of MMC 4 mg, two times a week for immediately postoperative three weeks, combined with postoperatively prolonged oral administration of Tegaful 800 mg/day more than three months was also significantly effective, especially in the cases of Dukes C stage (52.3% vs 40% in survival rates of 8 year). However, the analysis of recurrence did not prove that the intra-operative local intra vessel administration of MMC 10 mg was useful for the prevention of liver metastasis in colon cancer or pelvic recurrence in rectal cancer respectively.

[Comparative clinical study of adjuvant postoperative chemotherapy (5-FU, Tegafur, 5-FU + MMC) in curatively resected cases of gastric cancer. Study Group on 5-FU Oral Adjuvant Chemotherapy in Gastric Cancer].

A multi-center collaborative study was conducted in curatively resected gastric cancer patients at Stages II and III to compare oral 5-FU (Group A), oral Tegafur (Group B) and i.v. MMC + oral 5-FU (Group C). From May 1982 to April 1985, 1,012 cases were enrolled at 55 institutions. Some 138 (13.8%) were excluded, and 874 were analyzable. In the analysis of background factors, Group B had more cases with tumor of large diameter and advanced Stage. Adverse effects were relatively mild in all groups, and there was no problem in drug tolerance. Five-year survival rate was 67.6%, 62.4% and 68.6% in Groups A, B and C, respectively, reflecting no significant difference among them. It was 85.0%, 83.0% and 81.1% in Stage II and 52.5%, 51.0% and 59.0% in Stage III of Groups A, B and C, respectively. No significant difference was found, but Stage III of Group C showed a slightly higher survival rate. Supportive clinical study will be required to assess the usefulness of MMC as an introduction therapy. We found no difference in efficacy between 5-FU and Tegafur as maintenance therapy. The life-prolongation effect of fluoropyrimidines in comparison with surgery alone should be studied separately.

[Intramural injection of anticancer emulsion by gastroendoscopy].

Attempting to establish a topical chemotherapy for stomach cancer, injectable 5-FU emulsion (O/W type) has been developed. The distribution of 5-FU was analyzed by the procedures of quantification and X-rays. The combination of lipiodol having a contrast ability in the oil phase was studied. During the endoscopic examination before operation, 1.0 ml of 5-FU was injected at a site, a total of 50 ml at 2 to 4 sites in the submucosal tissue of the lesion. From the result of examination of 5-FU distribution in the extracted stomach and lymph node, it was confirmed that the drug was retained at the site of injection. In addition, 0.017-0.056 microgram/g on the average of 5-FU concentration was transferred to the secondary lymph node. At the same time, its distribution was monitored by X-rays. In those cases which received intermittent administration for twice or more before operation, the 5-FU concentration was detected at a higher rate than in cases receiving a single dose. This suggests that the therapy is markedly effective for stomach cancer and its metastasis to the lymph node as topical therapy.

[Study of pre- and post-operative chemotherapy with oral 5-FU in patients with gastric cancer. Study Group of Pre- and Post-Operative Oral 5-FU in Gastric Cancer].

A cooperative study group consisted of 37 medical institutions evaluated the utility of pre- and post-operative chemotherapy with oral 5-FU by comparing with the historical controls of the patients treated with operation alone who were registered in Japanese Gastric Cancer List. Of 634 patients entered in this trial, 366 patients received curative operation at macroscopical stages II and III were studied as eligible cases. The relative 5-year survival at macroscopical stage II and III was 68.6% with 5-FU, significantly higher than 62.6% with operation alone (p < 0.05). Particularly, 5-year survival at macroscopical stage III was markedly different between 57.1% with 5-FU and 42.7% with operation alone. In order to make more effective comparison with the historical controls in this 1-arm trial, the background factors in 5-FU and control groups were matched in the ratio of 1:2 or 1:3 with respect to the following 4 factors: tumor site, S, N and macroscopical tumor type (by Kajitani's classification). As a result, the 5-year survival at macroscopical stages II and III was significantly higher with 5-FU (67.5%, 69.1%) than with operation alone (59.9%, 59.8%), and in particular this tendency was more marked at macroscopical stage III. These results suggest the usefulness of adjuvant chemotherapy in macroscopical stage III gastric cancer.

[Estimates of circulating breast cancer-associated antigen CA 15-3 as a monitoring marker in patients with breast cancer].

CA 15-3 is a newly developed tumor marker detected by breast tumor-associated antigen 115D8/DF3 and is being studied as a monitoring marker in breast cancer patients (pts), even though its sensitivity as a screening marker is not so high. The cut off value of CA 15-3 was set at 27 U/ml. We assayed the plasma CA 15-3 levels of breast cancer pts from June 1985 for the purpose of estimating it as a monitoring marker in comparison with CEA. In the monitoring of over 2,000 postoperative pts, 23 were discovered to have metastatic lesions. For prediction of recurrence, CA 15-3 was useful for 11 pts (48%), while CEA was useful for 8 pts (35%), and CA 15-3 or CEA were useful for 14 pts (61%). Although it was little useful for local recurrence, CA 15-3 was highly useful for the prediction of organ & bone recurrence in 7/11 pts (64%). With regard to monitoring of the clinical course of metastatic carcinoma of the breast, the levels of CA 15-3 were positive in 47/68 pts (69%), while in contrast CEA was positive in 42/68 pts (62%). The trend of CA 15-3 was also highly correlated with the clinical course. CA 15-3 thus appears to be a better marker, especially as a monitoring marker, than CEA for breast cancer. Additional research will be required on this marker, but it seems likely that CA 15-3 combined with CEA would provide better information for the monitoring of breast cancer patients.

[Relationship between proliferative activity, and tissue differentiation and invasive mode in human oral squamous cell and colorectal carcinomas analysed by PCNA immunostaining].

The proliferative activity of carcinoma cells is generally considered to relate to the degree of the malignancy of carcinoma tissues. In this study, the proliferative activity at the tumor-stromal border was studied in 17 cases of oral squamous cell carcinoma (OSCC) and in 30 cases of colorectal adenocarcinoma (CAC) by means of proliferating cell nuclear antigen (PCNA) immunostaining, to evaluate the correlation between proliferative activity and tissue differentiation or invasive mode at the tumor-stromal border. No statistical difference was detected between the PCNA labelling index (PI) and the tissue differentiation of both OSCC and CAC. A significant difference was demonstrated between PI and invasive mode in OSCC, suggesting that the invasive mode at the tumor-stromal border relate to the degree of the malignancy of carcinoma tissues. However, no significance was found between PI and invasive mode of CAC. In addition, no difference of PI was demonstrated between tissue differentiation or invasive mode, and vascular invasion or lymph node metastasis. Therefore, it seems likely that the invasive mode at the tumor-stromal border in CAC also has no significance in deciding the degree of the malignancy of carcinoma tissues.

[Indications for total gastrectomy combined with pancreaticosplenectomy in the treatment of middle gastric cancer].

Of 1725 patients with middle gastric cancer (M, MA, MC) on whom radical surgery was performed (except for absolute non-curative resection) from 1960 to 1984 at the Cancer Institute Hospital, 238 patients in whom total gastrectomy or proximal partial gastrectomy combined with pancreaticosplenectomy was carried out, were submitted for both clinico-pathological and prognostic examination. The following results were obtained: 1. There are few indications for total gastrectomy plus pancreaticosplenectomy in patients with middle gastric cancer in which the lesions are superficial or semi-superficial. 2. For patients in whom the cancer occupies portions of the greater curvature, on posterior wall of the stomach, the whole stomach, or is of a non-localized type, the metastatic rate of lymph nodes No (10) or lymph nodes No (11) is high, then total gastrectomy plus pancreaticosplenectomy should be considered. 3. Lymph nodes No (2), (4d), (4sb) (7) and (9) can be good indices indicating presence or absence of metastasis of lymph nodes No (10) and lymph nodes No (11). 4. Radical surgery as well as effective adjuvant therapy are required for gastric cancer with invasion of the serosa.

[Distribution of recurrent lesions after radical resection of cancer of the thoracic esophagus].

In 150 patients who got cancer recurrence after curative resection for cancer of the thoracic esophagus, the sites where recurrent lesions were clinically detected for the first time were examined. The distribution of recurrent lesions in patients who did not undergo neck dissection at the operation (group A) differed from the distribution in those who underwent neck dissection (group B). Cervical and/or upper mediastinal recurrence occurred in 49% of cases in group A and in 11% of group B. On the contrary, middle or lower mediastinal recurrence was more often in group B. The distribution of recurrent lesions varied depending on the state of lymph node metastasis detected at surgery. Cervical and upper mediastinal recurrence was much more frequent than hematogenic recurrence in cases without lymph node involvement in group A, while hematogenic recurrence was more frequent in cases with both mediastinal and abdominal lymph node metastasis. In the upper mediastinum, recurrence along the recurrent laryngeal nerves was most frequent and it was supposed to have developed from residual lymphatic metastases. In the middle and lower mediastinum, recurrent lesions were located around the left main bronchus and descending aorta, and cancer infiltration of the neighboring organs was frequent. Recurrence at the abdominal paraaortic nodes was observed mainly in cases with perigastric lymph node involvement.