Norimasa Mori

Correlation between dysmenorrheic severity and prostaglandin production in women with endometriosis

The role of prostaglandins (PGs) in dysmenorrhea of endometriosis is poorly understood. The relationship between dysmenorrheic severity and prostaglandin production was investigated in endometriosis. Slices of normal myometrium, adenomyosis, normal ovary and endometrial cyst were incubated. 6-Keto-PGF1 alpha (a metabolite of PGI2), TXB2 (a metabolite of TXA2), PGF2 alpha, and PGE2 concentrations of the incubation medium were measured by RIA. The results showed that 6-keto-PGF1 alpha production in adenomyosis and endometrial cyst were significantly higher than those in normal myometrium and ovary. A direct relationship between the degree of dysmenorrheic severity and PGs production in tissue in endometriosis was observed.

Radiotherapy for carcinoma of the uterine cervix by remotely controlled afterloading intracavitary system with high-dose-rate

Between January 1978 and June 1985 seventy-nine patients with carcinoma of the uterine cervix were treated by a remotely controlled afterloading system (RALS) with high-dose-rate irradiation following external beam therapy. It was given once a week with a dose per fraction of 6 Gy to point A, 3 to 6 times, using applicators equipped with 1 Ci 60Co bead in each ovoid and two 1 Ci 60Co beads in a tandem 1 cm apart. Five-year survival rates were 85, 68, and 51 percent in Stages I, II, and III, respectively. Recurrences were noted in 12 patients (20%) with a minimum follow-up of 2 years: 4 patients in the parametrium, 2 in the central area, 5 in both the parametrium and central areas, and 5 in the para-aortic lymph node. Complications were noted in 13 patients (22%): temporary rectal bleeding in 7, stenosis of the sigmoid in 2, recto-vaginal fistula in 1, ileo-sigmoidal fistula in 1, and perforation of the intestine in 2 patients. The treatment results of the high-dose-rate schedule for RALS appear to be comparable to those of conventional low-dose-rate systems reported by others. Therefore a fraction dose of 6 Gy to point A once a week by RALS should be considered to be practical, economical, and acceptable to patients.

Detection of human T-cell lymphotropic virus type 1 infection by the polymerase chain reaction using dried blood specimens on filter papers.

A simple method for detection of proviral DNA sequences of human T-cell lymphotropic virus type 1 (HTLV-1) was developed using dried blood specimens on filter papers. The whole blood was blotted onto the Guthrie paper. After the blood has dried, the blotted paper was punched out into small discs. The discs were then boiled to prepare the template for PCR (filter paper-PCR method). The filter paper-PCR method detected even a single HTLV-1-infected cell in three discs. The sensitivity of the filter paper-PCR method was equivalent to that of the method in which DNA was extracted with phenol and used as the template for PCR (DNA extraction-PCR method). In addition, DNA in the blotted filter paper was still utilizable as the template after the storage at 25 degrees C for at least 7 wk. A total of 53 clinical specimens from 30 seropositive and 23 seronegative individuals who were screened by particle agglutination (PA) test were analysed for HTLV-1 DNA by both PCR methods. Of 30 PA-positive specimens, 28 were also positive for HTLV-1 antibody by Western blot (WB) analysis, but two were indeterminate. The twenty eight WB-positive and one of the two indeterminate specimens were positive for HTLV-1 proviral DNA by both PCR methods. Of 23 PA-negative specimens, 22 were negative for HTLV-1 proviral DNA by both PCR methods. However, one PA-negative specimen was positive by both PCR methods. This patient was a 16-mth-old infant who was born to an HTLV-1 carrier mother and fed thereafter without her breast milk. In comparison to DNA extraction-PCR method, the sensitivity and specificity of the filter paper-PCR method was 100%, respectively.

Interleukin-1 β stimulates prostaglandin E2 and F2α synthesis in human ovarian granulosa cells in culture

To evaluate the roles of interleukin-1 (IL-1) in regulation of ovarian prostaglandin (PG) synthesis, we examined the effects of IL-1β on PGE2, PGE2α, 6-keto-PGF1α and thromboxane (TX) B2 synthesis in cultures of human ovarian granulosa cells. Granulosa cells were obtained from hyperstimulated follicles in patients undergoing oocyte retrieval for in vitro fertilization-embryo transfer (IVF-ET). IL-1β increased immunoreactive concentrations of PGE2 and PGF2α in culture medium in time- and dose-dependent manners. Concentration of PGE2 was significantly higher after 24 h incubation with 5 or more units/ml of IL-1β, when compared to the control value obtained without IL-1β (P < 0.05). Concentration of PGF2α was significantly higher after 8 h incubation with more than 2 units/ml of IL-1β (P < 0.05). The increase in PGE2 was observed even in the presence of human chorionic gonadotropin (hCG), and blocked by indomenthacin, an inhibitor of cyclooxygenase. During a 10 day incubation period, stimulatory effects of IL-1β on PG synthesis were observed only on the first 2 days incubations. Concentrations of 6-keto-PGF1α and TXB2 were below our measurement limits. This study demonstrated that IL-1β stimulates PG synthesis in human ovarian granulosa cells in vitro. IL-1 seems to play an important role in regulating ovarian functions.

Cell-Mediated Immunity in Human Pregnancy

The function of thymus‐dependent lymphocytes (T lymphocytes) was studied in women during pregnancy and labor and postpartum by evaluating the blastogenesis of peripheral lymphocytes, which were stimulated with phytohemagglutinin‐P (PHA‐P) in both whole‐blood semimicroculture and purified lymphocyte culture. Data from 353 random samples (203 women) and 50 serial specimens from 10 women revealed that PHA‐P induced‐lymphocyte blastogenesis was significantly (p<0.005) reduced during pregnancy and labor but rapidly returned to normal several days after artificial termination in the early stage of pregnancy as well as after full‐term delivery. These results indicate that the T‐lymphocyte function in maternal peripheral blood is depressed by causes related to pregnancy. It seems very likely that depressed T‐lymphocyte function during pregnancy is caused by inhibitory factors in the blood plasma derived from the feto‐placental unit. Questions relating to the inhibitory factors in maternal plasma are discussed.

Synergistic effect of progesterone on prostaglandin E modulation of the mitogenic response of human peripheral lymphocytes

The combined effects of prostaglandins of the E series (PGE) and progesterone (P), both of which are found to increase locally in the secretory endometrium, were studied in mitogen-stimulated cultures of lymphocytes from human peripheral blood. When added separately to lymphocyte cultures, both PGE and P produced a concentration-dependent inhibition of T-cell, but not B-cell, mitogenesis. If added together, these agents caused a much greater inhibition of T-cell mitogenesis, with marked synergy, than that observed with either agent alone. The synergistic inhibition was achieved with endometrial concentrations of PGE and P. The kinetics of PGE- and P-mediated inhibition investigated at various times during lymphocyte activation indicated that both agents affected the early events of T-cell mitogenesis. PGE- and P-inhibition of T-cell mitogenesis were shown to be reversible phenomena by washing the cells treated with these agents. These results suggest that elevated levels of PGE and P in the secretory endometrium, acting synergistically in inhibiting the proliferative response of T cells, may participate in facilitating implantation of histoincompatible fetal tissue in the maternal uterus as non-specific local immunosuppressive factors.

Leiomyosarcoma of the vagina

A 46‐year‐old woman, who had very interesting tumor of the vagina microscopically, was presented. The removed tumor (40 g) in the upper lateral wall of the vagina was composed of two nodules. The cut surface of one nodule (20 g) was separated into two parts and they were diagnosed as necrosis and borderline case of leiomyomata microscopically. The other nodule (20 g) was diagnosed as leiomyosarcoma with interlacing bundles of spindle‐shaped cells with large hyperchromatic, irregularly shaped nuclei and abundant mutases. These bindings suggest a possibility that leiomyoma of the vagina may undergo malignant change to become leiomyosarcoma.

Purification of an NAD+-dependent 15-hydroxyprostaglandin dehydrogenase from the human placenta.

We purified 15-hydroxyprostaglandin dehydrogenase (PGDH) from the human placenta by gel filtration high performance liquid chromatography (GFHPLC) with spectrophotometric assay for the chromophore of 15-keto PG. Purification steps were as follows; homogenization and centrifugation at 10,000g, for 30 min, ammonium sulfate precipitation, DEAE-cellulose (DE-52), Sephadex G-100 gel filtration, NAD-affinity chromatography (AGNAD type 1), and GFHPLC. The specific activity was increased from 1.5 X 10(-2) unit/mg in the initial supernatant to 1272 X 10(-2) after GFHPLC. PGDH showed a single band on SDS-polyacrylamide gel electrophoresis and the molecular weight determined by GFHPLC and electrophoresis was approximately 50,000 daltons. After 4 months of storage, PGDH in a buffer containing 50% glycerol at -20 degrees C retained 80% of its activity. Michaelis constant for PGE2 was 0.4 microM at pH 7.0 with NAD as a cofactor. These properties of PGDH were compared with the results reported in other literature.

Eicosanoids production in endometriosis

In order to investigate the production of eicosanoids in human endometrium, myometrium, leiomyoma, adenomyosis, normal ovary, non-endometrial cyst and endometrial cyst, slices of each tissue were incubated. 6-Keto-prostaglandin (PG) F1 alpha, thromboxane (TX) B2, PGF2 alpha and PGE2 concentrations in the incubation medium were measured by direct RIA. 6-Keto-PGF1 alpha production of adenomyosis was significantly higher than that of endometrium, myometrium and leiomyoma, especially in the menstrual phase. The production of eicosanoids in endometrial cyst was significantly higher than that of non-endometrial cyst and normal ovary. These results suggest that endometriosis is associated with increased eicosanoid production in vivo.

Prostacyclin producing activity of human umbilical blood vessels in adrenergic innervated and non-innervated portions

The present experiment was performed in order to clarify the significance of prostacyclin (PGI2) in the regulation of human umbilical blood flow. Distribution of adrenergic nerve fibers in umbilical cord was examined by means of a modification of the glyoxylic acid fluorescence histochemical technique. PGI2 producing activity in various portions of umbilical blood vessels was measured by platelet bioassay. Adrenergic nerve fibers were observed only in the region surrounding umbilical arteries at the fetal end of the cord. PGI2 producing activity of umbilical arteries was significantly lower in the innervated region than in the non-innervated region. There were no significant regional differences in umbilical vein which has no adrenergic innervation. The relationship between vascular PGI2 producing activity and adrenergic innervation, and the significance of PGI2 in the regulation of human umbilical blood flow are discussed.