Frost-heave phenomena of 4He on porous glasses.
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Journal Article↗
Biomedical subjects
M Hiroi
Publications and source records attributed to M Hiroi.
The measurement of hyaluronidase activity in human spermatozoa by substrate slide assay and its clinical application.
Acrosomal hyaluronidase activity of individual sperm can be detected by a halo formation around the sperm head on hyaluronic acid substrate slides. The following results were obtained by this method in clinical practice. A significant correlation was found between hyaluronidase activity and sperm concentration in male infertility. Hyaluronidase activity increased as the concentration of sperm increased, and the least hyaluronidase activity was determined at less than 10 X 10(6)/ml. The group with more than 40% motility had a higher hyaluronidase activity than other groups with poor motility. Although there was no significant correlation between hyaluronidase and cumulus dispersion and fertilization rates in immature oocytes, there was an excellent correlation in mature oocytes in human in vitro fertilization. These findings suggest that a newly developed assay will be useful for evaluating sperm fertilizing capacity.
Acrosome↗
The development of porcine zona pellucida using monoclonal antibodies: I. Immunochemistry and light microscopy.
We established three monoclonal antibodies (Mabs) against the zonae pellucidae (ZP) of porcine oocytes, named STA-1, STA-2, and STA-3, and eventually we determined that they all reacted with the isolated ZP. Based on Western blotting without 2-mercaptoethanol (2-ME), STA-1 reacted with the 80,000-110,000 Mr component, STA-2 with the 42,000-63,000 Mr component, and STA-3 with the 40,000-80,000 Mr component of ZP. We immunohistochemically specified the components of porcine ZP reactive with the three Mabs during the course of follicular development. Each Mab reacted with both the ZP and the interfollicular cell space (IFCS). One ZP component, reactive with STA-2 and STA-3, was first produced in the primordial follicle and was not found at the cumulus follicle stage, which corresponds to the stage of large antral follicles more than 5 mm in diameter. Another ZP component, reactive with STA-1, was not produced until the secondary follicle stage, and was never found at the antral follicle stage. These results suggest that each ZP component is produced and secreted at a specific stage or stages of folliculogenesis.
Animals↗
The development of porcine zona pellucida using monoclonal antibodies: II. Electron microscopy.
Following our previous study on the immunohistochemistry of porcine zonae pellucidae (ZP), we undertook the present study to localize the components of the ZP with immunoelectron microscopy, using three types of anti-porcine-ZP monoclonal antibodies (Mabs), named STA-1, STA-2, and STA-3. Some organelles of the oocyte were seen to react with STA-2 and STA-3 prior to ZP formation. As soon as a follicle began to mature, STA-2 and STA-3 reacted with the perinuclear space and the endoplasmic reticular membrane of the oocyte. The follicle first reacted with STA-1 at the secondary follicle stage. At this stage, the positive reaction involved the follicular cell layer as well as the oocyte and ZP. Positive reaction was scattered within and limited to the interfollicular cell space and was never found in the cytoplasm of follicular cells. At the antral follicle stage, the oocyte was surrounded by a thick, electron-dense ZP. A strong reaction was observed in the outer layer, but no significant reaction occurred in the inner layer. The convex and ragged outer margin of the ZP was characterized by the strongest reaction.
Animals↗
Estrogen-binding protein in blood and follicular fluid, and its biochemical properties in human females.
Testosterone-estradiol binding globulin (TeBG) has been known to have specific binding to estradiol (E2) in blood, however an unknown binding protein having higher affinity to E2 than TeBG is thought to exist in blood. Therefore blood serum and follicular fluid were collected in normal females. Ammonium sulfate precipitation showed different maximum bound ratio between E2BP and TeBG. Concanavalin A (Con-A) Sepharose adsorption analysis showed that the Con-A-bound phase demonstrated estradiol-binding protein (E2BP). Isoelectric focusing showed TeBG in pH 4.9 and E2BP in pH 3.9. The substance giving the peak at pH 3.9 has a high affinity to dehydroepiandrosterone sulfate and E2, but the substance with peak at pH 4.9 shows a high affinity to 5-dihydrotestosterone and testosterone. E2BP changed concomitantly with total E2 in blood in the menstrual cycle, but in follicular fluid E2BP was found only in small amounts. It consisted of two components, one protein with low affinity binding and the other with high affinity binding. These results suggest that E2BP exists as a specifically bound fraction to E2, while TeBG, as a nonspecifically bound fraction to E2, is not directly involved in the E2-related biological function.
Adult↗
45,XO/46,X,dic(Yq) mosaicism in Turner's phenotype with endodermal sinus tumor of the ovary.
A case of a endodermal sinus tumor of the ovary in a patient with 45,XO/46,X,dic(Y) mosaicism is reported because of the rarity of the karyotype and condition. A 15-year-old girl was admitted to Yamagata University Hospital because of constipation for several days. Physical examination showed webbing of the neck, cubitus valgus and short stature. Her abdomen was bulging. Chromosomal analysis showed 45,XO/46,X,dic(Yq) mosaicism in karyotype. alpha-Fetoprotein and CA-125 in the serum were high. A left ovarian tumor was found by laparotomy; however, the right ovary was a streak gonad and the uterus was hypoplastic. An endodermal sinus tumor was diagnosed by a pathologist. After operation, cisplatin-vinblastin-bleomycin chemotherapy was instituted and the tumor marker went down. This patient is still healthy and under observation at the outpatient clinic.
Adolescent↗
Further evidence that prolactin controls the prepubertal sexual development in the female rat.
To assess the role of prolactin (PRL) in the puberty onset and the ovarian aromatase activity, hyper- and hypoprolactinemic conditions were induced by treating chronically prepubertal female rats with PRL and bromocriptine, respectively. Injection of rat PRL (5 micrograms, twice daily) from day 20 onward advanced the day of vaginal opening. By contrast, the suppression of PRL release by bromocriptine, which was provided in the drinking water at a concentration of 40 micrograms/ml, caused a significant delay in the age of occurrence of vaginal opening. This bromocriptine-induced delayed puberty was prevented by the concomitant administration of exogenous PRL, suggesting that the effect of bromocriptine is exerted through its suppressive effect on the endogenous PRL release. The in vitro release of estradiol by the ovaries from 30-day-old rats treated with PRL was found to be higher than that from the control animals, when the ovaries were perifused for 6 h with an excess of androstenedione. On the other hand, the ovaries perifused in the presence of androgen produced less estradiol in the bromocriptine-treated rats than in their controls. It appears, therefore, that PRL is more likely related to the aromatase activity in the maturing ovary. These results suggest that PRL may be one of the factors that regulate the timing of puberty onset by influencing the aromatase activity in the ovary.
Animals↗
[Combination chemotherapy of ovarian cancer with cisplatinum, aclarubicin and tegafur].
Two hundred fifty-nine patients with ovarian cancers were treated with the combination of CDDP, aclarubicin and tegafur. Ninety-seven of them had macroscopic diseases during the combination chemotherapy, and the overall anti-tumor response rate (CR + PR) for these patients was 54.6%. It was 60.0% for the 80 cases with epithelial ovarian cancer among them. The median survival time for the antitumor responders was 18 months, against 7 months for the non-responders and 18 months for 103 patients with advanced ovarian cancer (stage III-IV).
Aclarubicin↗
Skeletal muscle blood flow, metabolism and morphology in chronic congestive heart failure and effects of short- and long-term angiotensin-converting enzyme inhibition.
Blood flow to skeletal muscle during exercise is limited in patients with chronic congestive heart failure compared with normal persons. This may, in part, be attributed to the inability of the vessels to dilate adequately. In addition, an abnormal skeletal muscle metabolism with oxidative capacity can be demonstrated in patients with chronic congestive heart failure. The latter may partially explain why vasodilators and inotropes that improve central hemodynamics, global leg perfusion or skeletal muscle blood flow during exercise do not increase oxygen uptake by working muscle after acute drug administration. Furthermore, increases in blood flow to skeletal muscle may be redistributed to inactive or less oxygen-dependent muscle fibers. Although acute angiotensin-converting enzyme inhibition usually neither exerts substantial improvement in exercise hemodynamics nor interferes with leg perfusion during exercise, skeletal muscle blood flow and oxygen uptake gradually increases with long-term angiotensin-converting enzyme therapy, leading to improved systemic oxygen consumption and exercise tolerance. Several factors, such as reduction in plasma and vascular angiotensin II and sympathetic nervous activity, increased prostaglandin levels and renal function may contribute to this beneficial long-term effect.
Angiotensin-Converting Enzyme Inhibitors↗
Existence and biochemical analysis of estradiol binding protein (E2BP) in normal menstrual cycle.
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Adult↗
Inhibition of in vitro fertilization by a monoclonal antibody reacting with the zona pellucida of the oviductal egg but not with that of the ovarian egg of the golden hamster.
A glycoprotein designated ZP-0 is secreted from the epithelial cells of the oviduct and is bound to the zonae pellucidae (ZPe) of the oviductal eggs. We have previously established a monoclonal antibody reacting with ZP-0, termed AZPO-8. In the present study, we have examined the effect of AZPO-8 on in vitro fertilization in the hamster. Treatment of the oviductal eggs with AZPO-8 at concentrations of more than 100 micrograms/ml resulted in: (1) the formation of "zona precipitation" on the outer region of the ZP; (2) inhibition of sperm binding to ZP; (3) the acquisition of resistance to protease treatment. Fertilization was inhibited by AZPO-8 in a dose-dependent manner. These results indicate that a monoclonal antibody (MoAb) to an additional component of ZP derived from the oviduct shows an inhibitory effect on fertilization in vitro.
Animals↗
A monoclonal antibody reacting with the zona pellucida of the oviductal egg but not with that of the ovarian egg of the golden hamster.
A monoclonal antibody (mAb) designated AZPO-8 was produced by hybridizing a mouse myeloma with spleen cells from BALB/c mice immunized with materials obtained from the hamster oviduct. With an immunofluorescence test, AZPO-8 reacted with the zona pellucida (ZP) of ovulated eggs in the oviduct (ZP-OVI) but not with the zona pellucida of eggs in the ovary (ZP-OVA). Using indirect enzyme immunostaining, this mAb reacted with epithelial cells of the oviduct, the uterus (especially the cervical epithelium) and the gastric mucosa, but not with other hamster tissues examined. The reactivity of antigen-positive tissues was abrogated by pretreatment of the tissues with periodic acid. Western blotting analysis revealed that AZPO-8 reacted with substances of broad molecular weight range, and the strongest reactivity was detected at a molecular weight of approximately 200,000 in both cases when extract of ZP-OVI or the hamster oviduct was applied on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel. AZPO-8 showed strong hemagglutination activity only to group A human red blood cells. These results indicated that (1) ZP-OVI had an antigen that was not detected on ZP-OVA, (2) ZP-OVI and the oviduct shared the same antigenicity, and (3) the antigenic determinant reactive with the mAb might be carbohydrate in nature. A possible role of this antigen in fertilization was discussed.
Animals↗
Interaction between opioid peptides and monoamines in the regulation of gonadotropin secretion in the female rat.
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Animals↗
Effects of methylnitrosourea on visualization of acridine orange binding to DNA in mouse lymphoma L-1210 cells.
The purpose of this study is to examine effects of methylnitrosourea (MNU) on electron microscopic visualization of acridine orange (AO) binding to DNA in mouse lymphoma L-1210 cells and to demonstrate alteration of the euchromatin/heterochromatin ratio by morphometry. [3H] uridine uptake into RNA molecule is inhibited and percentage of AO positive cells is decreased to approximately 20% of that of the untreated control cells by treatment of the lymphoma cells with MNU. When the MNU-treated cells are cultured in the presence of 3-amino-benzamide, a specific inhibitor of poly(ADP-ribose) synthetase, the depression in RNA synthesis is prevented, and percentage of AO positive cells as well as numbers of AO chromatin interaction products per single cell nucleus are also completely recovered as compared to those in the untreated cells. In the MNU-treated cells the decreased number of AO positive cells coincides with a reduced [3H] uridine uptake as well as with a lowered euchromatin/heterochromatin ratio. The results suggest that visualization of AO chromatin interaction products in the proliferating cells is related not only to RNA synthesis in the cells, but also to the euchromatin/heterochromatin ratio.
Acridine Orange↗
Effects of actinomycin D on localization of acridine orange chromatin interaction complex in rat astrocytoma C6 cells.
The purpose of this study was to examine the effects of actinomycin D on localization of acridine orange (AO) binding to DNA in rat astrocytoma C6 cells and to discuss briefly the significance of AO chromatin interaction products. Actinomycin D markedly inhibited 3H-uridine incorporation into RNA and the percentage of AO positive cells was reduced to approximately 40% of that of the untreated control cells, whereas no distinct decrease of 3H-thymidine incorporation was induced by actinomycin D. Electron microscopic radioautography combined with the AO ultracytochemistry revealed that silver grains indicating binding of 3H-actinomycin D are located mostly over the euchromatin portion near the segregated nucleolus and heterochromatin and that no or only a few AO chromatin complex was found in nuclei labeled heavily with 3H-actinomycin D. These results of the present study together with the results of the previous studies seem to indicate that AO might selectively bind to active or derepressed DNA template sites for DNA dependent RNA polymerase in the euchromatin portion of the cell nucleus.
Acridine Orange↗
Rapid immunochemical assay of pregnanediol-3-glucuronide in urine and its clinical application.
A rapid and simple technique for assessing ovulation and luteal function is needed in clinical practice. A sensitive immunoassay kit is presented here for clinical use. An immunoassay of pregnanediol-3-glucuronide (Pd-3G) in urine was established. The sensitivity of this assay was 0.4 mg/l of Pd-3G. The standard error range of the recovery test was within 0.8 mg/l, and the coefficients of variation of within-run and day-to-day precision test were within 4%. Cross reactions of various steroids were observed in free pregnanediol, pregnanetriol and progesterone, but their reactivity exhibited less than 4%. The Pd-3G/creatinine (mg/g) ratio was demonstrated as nearly constant over a 24-hour period. A significant rise of Pd-3G coincided with a luteinizing hormone surge and continued during the luteal phase in the ovulatory menstrual cycle. In an anovulatory cycle, Pd-3G was found to be under 4 mg/g creatinine. This Pd-3G direct-assay method could be widely used in infertility clinics.
Adult↗
Correlation between the follicular gonadotropin inhibitor and the maturity of the ovum-corona-cumulus complex.
We assessed the correlation between the maturity of the ovum-corona-cumulus complex, the gonadotropin inhibitor in follicular fluids (FFs), and culture media derived from the granulosa cells of these follicles. FFs were obtained from patients (n = 20) stimulated with clomiphene citrate and human chorionic gonadotropin. The percentages of the inhibition of the ovarian weight augmentation (71.5% +/- 4.4%) and the number of ova (91.3% +/- 3.7%) in the mice given injections of FF from follicles containing mature ova and pregnant mare serum gonadotropin (PMS) were markedly greater when compared with those given FF from follicles containing immature ova and PMS (weight, 33.7% +/- 6.4%; ova, 60.8% +/- 5.8%). The immature granulosa cell culture media of days 4 to 6 and the mature cell media of days 1 to 3 greatly inhibited ovarian responses to gonadotropin (percent inhibition, immature, weight, 27.2% +/- 6.7%, ova, 48.1% +/- 13.2%; mature, weight, 42.9% +/- 9.2%, ova, 37.4% +/- 7.3%). It appears that mature follicles contain more gonadotropin inhibitor than immature follicles, that mature granulosa cells secrete the inhibitor more during days 1 to 3 of the culture than during days 4 to 6, and that immature cells secrete more during days 4 to 6 than during days 1 to 3.
Animals↗
[An immunochemical assay of pregnanediol 3-glucuronide in urine and its variation in the peri-ovulatory period].
A rapid, simple and sensitive immunochemical assay of pregnanediol 3-glucuronide (Pd-3G) in urine was established. The standard error range of the recovery test was within 0.8 microgram/ml, and the coefficient variation of the within run and day to day precision tests were within 4%. Cross reactivity of various steroids exhibited less than 4%. A significant rise of Pd-3G (mg)/creatinine (g) coincided with the LH surge and continued during the luteal phase in the ovulatory menstrual cycle. In an anovulatory cycle, Pd-3G was found to be under 4 mg/g. This Pd-3G direct assay method could be widely used for assessing ovulation and luteal function.
Adult↗