Sorbicillinol, a key intermediate of bisorbicillinoid biosynthesis in Trichoderma sp. USF-2690.

In the course of our screening program for free radical scavengers from Trichoderma sp. USF-2690, we found an unidentified metabolite (1) that appeared by the method used for HPLC analysis. Metabolite 1 gradually decreased with the production of bisorbicillinoids and was easily missed during the general isolation procedure. The LC-ESI-MS (negative) analysis for 1 gave m/z 247 as the (M-1)- ion peak. The hydrolysis of synthetic 6-O-acetylsorbicillinol (+/- -2) by 0.05 M KOH and acetylation of product 1 in an aqueous solution indicated that the structure of 1 was (6S)-4-(2,4-hexadienoyl)-3,6-dihydroxy-2,6-dimethyl-2,4-cyclohexadien-1-one, designated sorbicillinol, a quinol that has been postulated to be important in bisorbicillinoid biosynthesis.

Novel nikkomycin analogues: inhibitors of the fungal cell wall biosynthesis enzyme chitin synthase.

A series of novel nikkomycin analogue inhibitors of the chitin synthase of fungal cell wall was synthesized and evaluated for their inhibitory activities. Among them, the compound having a phenanthrene group at the terminal amino acid was found to possess strong anti-chitin synthase activity.

Synthesis and structure-activity relationships of novel nikkomycin analogs: inhibitors of the fungal cell wall biosynthesis enzyme chitin synthase.

A series of novel nikkomycin analogs, which inhibited chitin synthase, the fungal cell wall biosynthesis enzyme, has been synthesized and evaluated their inhibitory activities.

A case of vulvar schwannoma.

Schwannoma is generally a solitary, nodular, benign tumor of a peripheral nerve sheath, and it rarely affects the external female genitalia. Its most-common locations are the head, neck, upper and lower extremities, posterior mediastinum, and retroperitoneum. We treated a 29-year-old woman with a vulvar schwannoma. A tumor of 2.5 cm in diameter was resected for treatment and biopsy. It was a capsulate tumor with a smooth surface, and was elastic in consistency. A histological examination revealed a swirling pattern with some palisading of spindle-shaped cells in H.E. stains. Immunohistochemical studies showed the tumor to be positive for S-100 protein and negative for desmin. As a result, the mass proved to be schwannoma originating from the vulva.

Visual findings and histologic diagnosis of pelvic endometriosis under laparoscopy and laparotomy.

OBJECTIVE: To assess the diagnostic value of visual findings in the diagnosis of endometriosis from the histological point of view. STUDY DESIGN: 212 specimens from 107 patients with benign (74.8%) or malignant (25.2%) disease were obtained by biopsy or resection under laparoscopy (65 patients) or laparotomy (42 patients). Ages ranged from 19 to 62 (mean age 36.4). Visual findings were classified according to the criteria established by the Endometriosis Committee of the Japan Society of Obstetrics and Gynecology in 1993. Specimens were stained with hematoxylin-eosin, and 15 cases with periodic acid-Schiff stain or silver impregnation stain. RESULTS: Among pigmented lesions, endometriosis was found in 73.0% of specimens from the pelvic peritoneum and in 56.4% of those from the ovaries. Blueberry spots in the pelvic peritoneum as well as ovarian chocolate cysts showed the highest positive rate. In the presence of multiple or complex pigmented lesions of the pelvic area, the rate was still higher (88.6%). Those rates were due to our inclusion of inaccurate and incomplete biopsy specimens. Endometriosis of nonpigmented lesions was found in only 11 patients (12.0%) who also had pigmented lesions and/or adenomyosis. CONCLUSION: The laparoscopic diagnosis of endometriosis can be made only when multiple complex pigmented lesions are observed, but, otherwise, histopathological confirmation is necessary.

Morphologic hysteroscopic criteria suggestive of endometrial hyperplasia.

OBJECTIVES: To evaluate the morphologic hysteroscopic criteria leading to a diagnosis of endometrial hyperplasia and compare their accuracy with that of histology. METHODS: A total of 95 hysteroscopic examinations were evaluated. Of these, 37 had a histologic diagnosis of normal endometrium and the remaining 58 of simple or complex endometrial hyperplasia. We compared the morphologic hysteroscopic criteria for the two groups using Pearson's chi-squared and Fisher's exact test. RESULTS: Only the presence of endometrial glands presenting a cystic pattern at hysteroscopy gave statistically significant results (P < 0.05), with low sensitivity (15.79%), high specificity (97.29%) and a relative risk of 6.75. With a prevalence of endometrial hyperplasia of 22.97% in a population of women with metrorrhagia, the positive predictive value was 63.53% and the negative predictive value was 79.40%. CONCLUSION: Additional, prospective studies are needed to determine the real value of the diagnostic morphologic parameters under consideration.

Endocrinological and histological changes after treatment of uterine leiomyomas with danazol or buserelin.

The following results were obtained from our investigations performed to compare the effects of danazol (400 mg/day orally for 4 to 6 months, n = 164) with those of buserelin (900 micrograms/day intranasally for 4 to 6 months, n = 83) on patients with uterine myomas. The effectiveness of buserelin (76.1%) in shrinking uterine myomas was better than that of danazol (56.7%). The serum-estradiol and progesterone levels were lower in the buserelin group than in the danazol group. The estrogen and progesterone receptors in a hysterectomied uterus were positively stained even at the end of those therapies. The degree and frequency of hyaline degeneration were higher in the buserelin group than in the danazol group. Thus, the shrinking of uterine myomas and the degree of hyaline degeneration might be caused by the lowering of uterine-tissue hormones, particularly of estradiol levels.

Endocrine function of the peri- and postmenopausal ovary.

The concentration of sex steroids in peripheral and ovarian venous blood was measured in 130 perimenopausal women. We found that the postmenopausal ovary continues to secrete a large amount of testosterone and a moderate amount of androstenedione, estrone, and dehydroepiandrosterone. Although the mean estradiol levels in the ovarian vein of postmenopausal women was low, a few subjects showed estradiol levels higher than 50 pg/ml. Postmenopausal ovaries probably continue to secrete estradiol under some endocrine conditions. These data show that the postmenopausal ovary maintains some secretory function related to estrone and estradiol. However, the endocrine effect of the postmenopausal ovary may be related to its contribution of androgens to the plasma pool of estrogens through extragonadal conversion.

Hereditary abnormality of luteinizing hormone resulting in discrepant serum concentrations determined by different assays.

We herein report familial defects in the molecular structure of luteinizing hormone (LH). The propositus was a 29-year-old women with repeated abortion in whom the serum LH concentration was extremely low determined by an immunoradiometric assay utilizing two monoclonal antibodies for the intact LH dimer and beta-subunit of LH (SPAC-S kit). Further studies on the serum LH concentration in this propositus by five different assay systems gave various results ranging from low to normal values. Bioassay of the propositus's LH using C57 black mice showed normal in biological activity. These data suggest that LH in the propositus is abnormal in terms of its molecular structure located in the bond region between alpha and beta subunits. A family study showed that the bioactivity of LH was normal in all family members. The serum LH was not detected in either the propositus or her brother with the SPAC-S kit even after the administration of LHRH, while the serum LH concentrations in the father, mother and sister were approximately half the normal range, indicating that the defect is hereditary and the mode of inheritance is autosomal dominant: the propositus and brother were homozygously affected, and the parents and sister heterozygously affected.

[Long-term administration of carmofur as a post-operative adjuvant chemotherapy for cervical adenocarcinoma. Cervical Adenocarcinoma Cooperative Research Association].

Cervical adenocarcinoma and adenosquamous cell carcinoma are low in radiation sensitivity, and the prognosis is said to be inferior to that of squamous cell carcinoma. Eleven facilities nationwide participated in the historical control study on the recurrence prevention effect of carmofur (HCFU) administered at 300 mg/day for more than two years postoperatively as an adjuvant chemotherapy to patients with cervical adenocarcinoma or adenosquamous cell carcinoma for which radical operation is possible. Registered for the study was a total of 252 patients: 77 patients for whom administration of carmofur was begun during the period from January 1987 and March 1989, 71 patients (control 1) who were treated for the cancer after January 1982 but did not receive adjuvant chemotherapy, and 104 patients who received adjuvant chemotherapies with other than carmofur (control 2). In analyses with the Kaplan-Meier method, the carmofur administration group demonstrated a better cumulative survival rate and disease free rate than control 1 (no adjuvant chemotherapy), suggesting carmofur was effective in preventing the recurrence of cervical adenocarcinoma and adenosquamous cell carcinoma.

Cyclic bcl-2 gene expression in human uterine endometrium during menstrual cycle.

To investigate an apoptotic role for bcl-2 in menses, we studied bcl-2 protein production throughout the cycle. Bcl-2 protein was observed in endometrial glandular, stromal and myometrial smooth-muscle cells. Glandular cells expressed bcl-2 at proliferative through early secretory phases but not at late secretory through menstrual phases. This cyclic expression of bcl-2 gene was similar to that of oestrogen and progesterone receptors. Thus bcl-2 expression in glandular cells may be regulated by ovarian hormones. The disappearance of bcl-2 expression in glandular cells at late secretory phase was consistent with the appearance of apoptotic cells in the same phase.

[Correlation of ill-defined syndrome with depression in the climacterium].

The psychologic and somatic symptoms at climacterium are multiplicative, ill-defined, and nonspecific. We studied 390 climacteric patients with ill-defined psychologic and somatic symptoms. While such symptoms arise from an estrogen-deficient state of the menopause, these also occur in certain major psychiatric disorders, especially those of depression. Thirteen and twenty-one percent of the women were diagnosed as depression and masked depression, respectively. The incidence of generalized and panic disorder was 6.7% in each. The ill-defined syndrome of the menopausal age may be severe and persistent enough to meet the diagnosis of a major psychiatric illness, i.e. depression. As these disorders are important differential diagnoses, personality analysis and careful observation of clinical course may be indicated. Furthermore, the psychosomatic approach is recommended for all of these cases.

Antigenic alteration of an anomalous human luteinizing hormone caused by two chorionic gonadotropin-type amino-acid substitutions.

We analyzed the nucleotide sequence of the luteinizing hormone beta subunit (LH beta) in a patient with an anomalous LH. This anomalous LH showed abnormal immunogenicity, but normal bioactivity, suggesting that this variance of antigenicity was caused by amino acid substitution(s). In the anomalous LH, two single amino acid substitutions, Trp(TGG) to Arg(CGG) and Ile(ATC) to Thr(ACC), were found at the codon for the 8th and 15th residue of LH beta. These two substituted amino acid residues of the anomalous LH are identical to those of chorionic gonadotropin, but not to those of LH, although the rest of the region showed the normal sequence of human LH beta. Pedigree analysis by direct DNA sequencing revealed that the parents of the patient and the healthy sister were heterozygotes for the mutation and the patient and the healthy brother were homozygotes.

Conservative therapy for microinvasive carcinoma of the uterine cervix.

The purpose of this study was to evaluate conservative therapy for microinvasive carcinoma (MIC) of the cervix. Large contact Nd-YAG laser conization (laser cone) with contact vaporization using contact probes was performed on 90 cases of MIC and 21 cases of early invasive carcinoma (early IC), with long-term postoperative follow-up for patients at Osaka Medical College, Osaka, Japan. The cure rates of MIC and early IC were 96.7 and 80.9%, respectively. The rates of incomplete excision and true residuation in cases with MIC were 17.8 and 10.0% and those in cases with early IC were 76.2 and 19.0%. Therefore, the spontaneous cure rate following incomplete excision of MIC and early IC was 43.7 and 75.0%. Hysterectomy was performed, for various indications, after cone in 5 patients (5.6%) with MIC, none of the patients with IC of less than 3 mm, 2 (22.2%) of the patients with IC of 3.1 to 5 mm, and 7 patients (87.5%) whose IC was 5.1 mm or more in depth. Combining the data from our department and those in the literature collected an incidence of lymph node metastasis of 0.15% for MIC and of 1.13% of IC of less than 5 mm. In conclusion, conservative therapy is indicated for MIC, and the contact laser cone with contact vaporization is considered to be an excellent therapeutic conservative method.

Direct determination of NotI cleavage sites in the genomic DNA of adult mouse kidney and human trophoblast using whole-range restriction landmark genomic scanning.

Restriction landmark genomic scanning (RLGS) is a method for visualizing restriction landmarks, employing direct labeling of restriction sites of genomic DNA and high-resolution two-dimensional electrophoresis. We determined the conditions for both the first and second dimensions of RLGS that define all of the restriction fragments which carry the NotI landmark. Using this system, we determined the number of cleavable NotI sites of genomic DNA from the mouse kidney (C57BL/6) and from the human placenta. The mouse and human genomes were cleaved at 2,380 +/- 80 sites (4,760 +/- 160 spots) and 3,240 +/- 110 sites (6,480 +/- 220 spots), respectively with NotI.

Efficacy and short-term effects of pravastatin, a potent inhibitor of HMG-Co A reductase, on hypercholesterolemia in climacteric women.

The effects of pravastatin, a potent inhibitor of HMG-CoA reductase, on serum lipids and lipoproteins were studied in 86 patients (36 premenopausal and 50 postmenopausal women) suffering from primary hypercholesterolemia. The effects of this lipid-lowering drug on serum gonadotropins and sex steroids are also reported. Pravastatin produced a mean reduction in serum total cholesterol (T-CHO) of 24.7%, triglyceride (TG) of 30.0%, and low-density lipoprotein cholesterol (LDL-CHO) of 36.0% after treatment for three months. The drug also produced a mean increase in high-density lipoprotein cholesterol (HDL-CHO) of 1.4% after three months. Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estrone (E1), estradiol (E2), and testosterone (T) levels were not significantly changed from pre-therapy levels in either pre- or post-menopausal women after treatment for six months. Pravastatin is well-tolerated for six months and is a very effective lipid lowering agent with no effects on the biosynthesis of sex steroids. These findings suggest that pravastatin can be used for treatment of primary hypercholesterolemia in women around menopause.

Characterization of epidermal growth factor (EGF) receptor and biological effect of EGF on human uterine cervical adenocarcinoma cell line OMC-4.

In this paper, we investigated how epidermal growth factor (EGF) acts on growth and regulation of extracellular matrix components and their degenerative enzymes in uterine cervical adenocarcinoma cells in vitro. Effects of EGF on cell growth, DNA synthesis, and laminin, collagen IV, and tissue plasminogen activator (t-PA) production of human uterine cervical adenocarcinoma cell line OMC-4 were examined, together with the characteristics of its EGF receptors. Scatchard plot of EGF binding to OMC-4 indicated two classes of binding sites with a dissociation constant of 170 pM and 510 pM. The total binding sites were 1.6 x 10(5)sites/cell. The number of OMC-4 cells did not increase in the presence of EGF, whereas 3H-thymidine incorporation was inhibited by EGF at concentrations of 1 and 10 nM. The production of laminin and collagen IV by OMC-4 cells was inhibited by EGF, whereas that of t-PA was significantly promoted at the physiological concentration of 0.1 nM. These results suggest that EGF is closely associated with regulation of proliferation and extracellular matrix degradation of uterine cervical adenocarcinoma cells.

Plasma lipid and lipoprotein levels in perimenopausal women. Clinical research in 1198 Japanese women.

Hyperlipidemia is associated with an increased prevalence of cardiovascular and cerebrovascular disease. We studied plasma lipid and lipoprotein levels with a survey of endocrine changes in the perimenopausal Japanese women. Total cholesterol, triglyceride, and low density lipoprotein cholesterol levels increased with age, whereas high density lipoprotein cholesterol level fell slightly. The rise in triglyceride and low density lipoprotein levels were particularly more marked just before the onset of the menopause. It might be important that the rapid rise of triglyceride and low density lipoprotein levels at about the menopause age was associated with a decline of estrogen levels. The percentage of hyperlipidemia in the perimenopausal women who showed total cholesterol level exceeding 5.7 mmol/l (220 mg/dl) and low density lipoprotein cholesterol level exceeding 3.9 mmol/l (150 mg/dl) were 31.9% and 27.2%, respectively. It was concluded that we need to recognize relatively high incidence of hyperlipidemia in perimenopausal period, and need to pay careful attention to plasma lipid and lipoprotein levels for prevention of coronary heart disease in perimenopausal women.