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Biomedical subjects

M Irahara

Publications and source records attributed to M Irahara.

At least 91 records · Page 5Linked to original sources

Effect of 2-buten-4-olide, an endogenous suppressant of feeding, on reproductive function in rats.

The effects of 2-buten-4-olide, an endogenous feeding suppressant, on the estrous cycle and LH secretion were studied to determine the influence of this compound on reproductive function. Estrous cycling female Wistar rats were treated ip with 2-buten-4-olide (0, 30 or 100 mg.kg-1.day-1) for 2 weeks. Treatment with 100 mg.kg-1.day-1 delayed the estrous cycle. 2-Buten-4-olide increased the pituitary content of LH (1651.3 +/- 164.4 vs 927.7 +/- 65.1 ng/pituitary; p less than 0.01), and decreased the serum LH level compared with the control level in diestrus (0.16 +/- 0.01 vs 0.26 +/- 0.03 microgram/l; p less than 0.05). However, it did not affect the GnRH content of the mediobasal hypothalamus. The direct effects of 2-buten-4-olide on the pituitary response to GnRH was examined by perifusing the pituitary. Medium containing 2-buten-4-olide (10(-4) mol/l) suppressed the pituitary response to GnRH (2 micrograms/l) (percent increase at 50 min after start of GnRH stimulation: 180 +/- 47 vs 406 +/- 66%; p less than 0.05). These findings suggest that 2-buten-4-olide is involved in the regulation of pituitary gonadotropin secretion directly by suppressing the pituitary responsiveness to GnRH, and that 2-buten-4-olide may play an important role in starvation-induced anestrus.

4-Butyrolactone↗

Significance of D-mannose as a sperm receptor site on the zona pellucida in human fertilization.

The role of monosaccharides in human fertilization was studied by testing their effects on penetration of spermatozoa into mature human oocytes (zona penetration test). When oocytes were pretreated with concanavalin A, wheat germ agglutinin, or Ricinus communis agglutinin-I at a concentration of 100 micrograms/ml, no spermatozoa were found to bind to or penetrate through the zona pellucida. Penetration of spermatozoa was restored when the zona pellucida pretreated with wheat germ agglutinin and Ricinus communis agglutinin-I were rinsed with N-acetyl-D-glucosamine (wheat germ agglutinin inhibitor) and D-galactose (Ricinus communis agglutinin inhibitor), respectively. Conversely, the blocking effect of concanavalin A on sperm penetration was not reversed by treatment with D-mannose (concanavalin A inhibitor). Furthermore, pretreatment of spermatozoa with D-mannose (50 mmol/L) completely inhibited sperm penetration through the zona pellucida. However, sperm penetration was clearly demonstrated when the zona pellucida was pretreated with D-mannose. These data suggest that D-mannose residues are essential in, or sterically closely related to, the sperm receptor site on the human zona pellucida.

Carbohydrates↗

Treatment of malignant ascites with allogeneic and autologous lymphokine-activated killer cells.

Two cases of peritonitis carcinomatosa with gynecological cancer, which did not respond to conventional treatment, were treated by intraperitoneal adoptive immunotherapy with allogeneic or autologous lymphokine-activated killer (LAK) cells plus interleukin-2. In case 1, reduction of acute ascites and disappearance of malignant cells from the peritoneal fluid and decreased levels of tumor markers (CA12-5 and CA19-9) were demonstrated during the treatment. In case 2, which also received additional treatment for pleuritis carcinomatosa, reduction of ascites and pleural effusion and disappearance of malignant cells from the pleural fluid were noted. In these cases allogeneic LAK therapy was well tolerated. This therapy did not prolong the patients' lives, but the findings indicate that it had appreciable local antitumor effects on peritonitis and pleuritis carcinomatosa in these patients with gynecological cancer.

Ascites↗

[Clinical implication of serum antibody to Thomsen-Friedenreich antigen in patients with gynecological malignancies].

Thomsen-Friedenreich antigen (T-antigen) is a carbohydrate antigen that is expressed in a variety of cancer tissues. T-antigen is thought to have an antigenicity because circulating T antibodies can be detected as natural antibodies in humans. In this study, we examined the serum T antibody titers in patients with gynecological cancer using the hemagglutination test, and studied the relationship between the expression of T-antigen in cancer tissues and serum TA-4 levels and serum T antibody titers. Serum T antibody titers in patients with gynecological cancer were lower than those in normal controls, especially in endometrial and ovarian cancer (p less than 0.05) in which T-antigen was strongly expressed. Furthermore, the low antibody titers correlated with the expression of T-antigen in cancer tissues. T antibody titers significantly increased (p less than 0.01) after operation and the inverse relationship was found with the levels of circulating TA-4 in cervical cancer patients. These findings suggests that patients with gynecological cancer immunologically responded to T-antigen and the measurement of circulating T antibodies may be useful as an indicator of the progression of cancer in tissues.

Antibodies↗

Inhibitory effects of tunicamycin on the mixed lymphocyte reaction and mitogen-induced lymphocyte blastogenesis.

The role of asparagine(Asn)-linked saccharides on the surface of lymphocytes in cellular interactions was examined by performing studies on the effects of tunicamycin (TM), which inhibits the glycosylation of proteins N-glycosylated at asparagine residues, on the mixed lymphocyte reaction (MLR) and mitogen-induced lymphocyte blastogenesis of human lymphocytes by measuring 3H-thymidine (TdR) incorporation. Responses were expressed as percentages of that of the control (MLR without TM). The lymphocyte blastogenesis in the one- and two-way MLR were, respectively, 43.1% and 48.0% of the control at 0.1 microgram/ml of TM, and 5.5% and 7.2% at 1 microgram/ml of TM. The inhibitory effect of TM on the one-way MLR was shown using TM-pretreated stimulator cells, TM-pretreated responder cells or both. TM blocked lymphocyte blastogenesis in the secondary as well as in the primary MLR. The inhibitory effect of TM on the two-way MLR was observed when TM was added on day 0 to day 2, but not on day 4 of incubation. TM blocked mitogen-induced lymphocyte blastogenesis by phytohemagglutinin, concanavalin A or by pokeweed mitogen. As TM had no cytotoxic effect on cultured cells, these inhibitory effects of TM were thought to be due to the loss of Asn-linked saccharides from glycoprotein of the surface of lymphocytes. These findings indicated that Asn-linked saccharides of glycoprotein on the surface of lymphocytes were important in cellular interactions that are necessary for the cellular immune response.

Antigens, Surface↗

[Expression of Thomsen-Friedenreich antigen (T-Ag) in gynecologic cancer tissues].

The expression of Thomsen-Friedenreich antigen (T-Ag), the precursor of MN blood group antigen, was studied by the ABC method with Arachys Hypogaea lectin (PNA) and the relationship of the results to the histology and clinical behavior of primary gynecologic cancers were examined. The results were as follows: 1. In cervical squamous cell tumors, the incidence of the expression of T-Ag in invasive cancers (52.9%, 37/70) was significantly higher (p less than 0.05) than that in intraepithelial tumors (28.6%, 8/28). No cryptic T-Ag(-) tissue was found in intraepithelial tumors or microinvasive cancers. 2. In squamous cell carcinomas of the cervix, the incidence of cryptic T-Ag(-) tissue was high (36.4%, 4/11) in tissue of the small cell non-keratinizing type, the most undifferentiated type, whereas it was not found in 5 cases of well-differentiated keratinizing carcinoma examined. 3. In stage Ib and II cervical cancers, no relationship was recognized between the expression of T-Ag in primary lesion and extension to the parametrium or the pelvic lymph nodes. 4. The incidence of T-Ag(+) tissue in adenocarcinomas of the uterine cervix (62.5%, 5/8), the endometrium (91.7%, 11/12), and the ovarian (80%, 8/10) was higher than that in squamous cell carcinoma of the uterine cervix (52.5%, 32/61). 5. In endometrial cancers, no relationship was recognized between the expression of T-Ag and hormone receptor status.

Adenocarcinoma↗

Differences in clinical significance of blood group antigens A, B, and H in carcinoma tissue in the uterine cervix.

The losses of blood group antigens A, B, and H in carcinoma tissue of the uterine cervix were studied by the avidin-biotin-peroxidase complex (ABC) method and the relations of these losses to invasion and dedifferentiation of primary cancer were examined. The incidence of cases showing complete loss of A or B antigen increased in proportion to the progression of cancer, but in most cases even of invasive cancer, H antigen, the precursor of A and B antigens, was detected. Complete loss of H antigen was not demonstrated in well-differentiated keratinizing invasive carcinomas, but was seen in 15% (15/101) of the cases of large cell non-keratinizing type cancer and 50% (8/16) of those of small cell non-keratinizing type cancer. No relationship was found between losses of A, B, and H antigens and parametrial spread of carcinoma or metastasis to the pelvic lymph nodes, but the incidence of death within 2 years after hysterectomy was higher in H antigen-negative cases than in H antigen-positive cases. These results indicate that loss of A and B antigens depends on some activity of invasion of cancer, while loss of H antigen strongly indicates dedifferentiation of cancer cells and also may indicate a poor prognosis.

ABO Blood-Group System↗

Blocking of human fertilization in vitro by sera with sperm-immobilizing antibodies.

Serum samples with sperm-immobilizing antibody activity from six women were examined for ability to block sperm-egg interaction by a zona penetration test where human follicular ova matured in vitro were used. Exposure of spermatozoa from a fertile healthy donor to the sera impaired binding to and penetration through the zona pellucida of the spermatozoa completely in five cases and incompletely in one case. Successful fertilization in vitro was achieved by using fetal cord serum instead of autoserum of the patient included in the in vitro fertilization and embryo transfer program. These results suggest that interference with sperm-egg interaction may be an additional mechanism of infertility that is caused by antisperm antibodies.

Adult↗

Antibody reactivity with porcine zona pellucida.

A comparative study on anti-zona antibody activities in the sera from clinically defined categories of patients registered at the WHO Reference Bank for Reproductive Immunology was performed at five different laboratories with different detection methods. Considerably higher incidences of positive reactions were detected by immunofluorescence on porcine zonae in infertile women (16.3%) than in control subjects (7.1%). A similar proportion of positives was found by radioimmuno-binding assay (RIBA) using porcine zona antigen preparation in the infertile group (13.0%) but not in the female control group (0%), giving an indication of the specificity of this test. It is noteworthy that high incidences of positives were observed by RIBA with sera from male subjects with unexplained sterility, vasectomy and aspermatogenesis. A test system of passive hemagglutination reaction (PHAR) using purified porcine zona substance as antigen gave a low but slightly higher incidence of positives in infertile (3.1%) than in control sera (0.9%). No positive reactions were observed with infertile and control sera by another PHAR or by radioimmunoassay using an antigen preparation common to the two test systems. Anti-zona activities in these sera were therefore seen to vary, depending largely upon the detection systems and the antigen preparations.

Animals↗

Inhibitory action of somatostatin on meiotic maturation of cultured porcine follicular ova.

To investigate the physiological importance of somatotrophin release-inhibiting factor (SRIF), effects in vitro of synthetic SRIF 14 on germinal vesicle breakdown (GVB) of cultured porcine follicular ova were studied. The proportion of ova with GVB decreased gradually and significantly with increasing concentrations of SRIF 14 in the range from 6 X 10(-12) to 6 X 10(-7) M during a 22 h period of culture. The inhibitory effect was apparent for the period between 14 and 22 h in the course of culture but was reversed by a concomitant addition of anti-SRIF to the medium. Neither synthetic oxytocin, vasoactive intestinal polypeptide nor substance P exerted any inhibitory or stimulatory action on GVB. These results suggest a limited but definite inhibitory action of SRIF on GVB of porcine ova.

Animals↗

Detection of anti-zona pellucida activities in human sera by the passive hemagglutination reaction.

A simple method of detecting anti-zona activities in human sera has been developed by passive hemagglutination reaction ( PHAR ) using bovine erythrocytes as indicator cells coated with purified porcine zona substance. By this test, 8 of 88 serum samples (9.1%) from infertile women gave positive reactions, whereas only 1 of 90 control sera (1.1%). A positive immunofluorescence (IF) assay confirmed the results of the PHAR in seven of the eight positive samples. Positive PHAR and IF on porcine zonae were abolished after adsorption with porcine erythrocytes, but IF on human zonae was retained by four of eight serum samples, indicating the presence of anti-zona autoantibodies in these infertile sera that were directed to an antigen(s) proper to human zonae.

Adult↗

Evidence for existence of somatostatin-like immunoreactivity with molecular heterogeneity in porcine ovaries.

Measurement of somatostatin (SRIF) by RIA in porcine ovarian tissues after removal of the follicular fluid revealed the existence of immunoreactive (IR) SRIF at concentrations of 2.2 +/- 0.6 ng/g wet weight (mean +/- SD). On gel chromatographic analysis of ovarian extracts one major and two minor components with SRIF-like immunoreactivities have been detected. The major component comprised 59.5 +/- 15.1% of the total IR-SRIF and was coeluted with synthetic tetradecapeptide (SRIF 14). The two other components seemed to have almost the same molecular weights as synthetic octacosapeptide (SRIF 28) (25.1 +/- 10%) and pro-somatostatin (15.4 +/- 5.5%), respectively. It is concluded that SRIF in porcine ovaries is heterogeneous, comprising SRIF 14 as a major component, as well as larger forms of SRIF which probably represent SRIF precursors.

Animals↗

The mechanism of the effect of combination treatment with clomiphene and bromocriptine in patients with normoprolactinemic anovulation.

The combination treatment with bromocriptine and clomiphene citrate was applied to 11 normoprolactinemic anovulatory patients who did not respond to clomiphene citrate alone. This combination treatment restored ovulation in 8 of these patients (72.7%). Conception was observed in 2 patients (18.1%) out of 11. The patients who responded to combination treatment showed a significant increase in the serum level of estradiol in the preovulatory phase, of progesterone in the mid-luteal phase, and a significant decrease of serum prolactin. They also showed significant increase in the frequency of luteinizing hormone (LH) pulsatility on day 12 of the cycle from 1.38 +/- 0.86 to 3.75 +/- 0.83 pulses/4h. The 3 patients who did not respond to combination treatment showed no increase in the serum level of estradiol or progesterone, but showed increase in the frequency of LH pulsatility in spite of continuous anovulation. These results indicate that the combination treatment with bromocriptine and clomiphene citrate is effective for treatment of patients with normoprolactinemic anovulation who do not respond to clomiphene alone, and suggest that the mechanism of the effect of combination treatment is related to an increase in the frequency of LH pulsatility caused by bromocriptine, which in turn stimulates follicular maturation.

Adolescent↗

Specific cytotoxicity of antibody to YAL-198, a sperm antigen peptide, to murine zygote.

Active immunization with the peptide segments rSMP-230 and YAL-198, corresponding to the hydrophilic extracellular domain of two human sperm antigens (rSMP-B and YWK-II, respectively), reduced fertility in female rats by different mechanisms. The anti-rSMP-230 antibody interferes with human and murine fertilization, and the anti-YAL-198 antibody blocks the development of mouse embryos. The authors examined in vitro at which stage the antibodies to rSMP-230 and YAL-198 were cytotoxic to murine embryos up to morula/blastocyst stage. Anti-rSMP-230 antibody was not cytotoxic to any stages. On the other hand, the anti-YAL-198 antibody arrested the growth of embryos at the 2-cell stage but not at more advanced developmental stages. When the anti-YAL-198 antibody was used, spotty staining was observed only on the surfaces of embryos that had arrested at the 2-cell stage. Unstained embryos, however, continued to develop normally. In contrast, the anti-rSMP-230 antibody stained murine sperm but failed to stain murine ova and embryos. The present results suggest that the human sperm components rSMP-B and YWK-II play important roles in sperm-egg interaction and early development of the embryo, respectively.

Animals↗

Progesterone accelerates the onset of capacitation in mouse sperm via T-type calcium channels.

This study was undertaken to evaluate whether progesterone induces capacitation of mouse spermatozoa. When sperm were evaluated by chlortetracycline staining, addition of progesterone significantly increased the proportion of spermatozoa exhibiting the B pattern at 60 minutes of incubation, compared with that before incubation (23 +/- 6.2% vs. 13 +/- 2.9%, p < 0.01) and that in hTF medium without progesterone (23 +/- 6.2% vs. 13 +/- 4.2%, p < 0.01). If the redistribution of proteins in sperm plasma membrane such as protein binding calcium ion were defined as capacitation, it could be said that progesterone promoted capacitation of mouse sperm. This progesterone-induced capacitation was prevented by depletion of extracellular calcium ion and addition of NiCl2, a T-type calcium channel blocker, although thapsigargin, an inhibitor of Ca2+-ATPase, did not increase the number of capacitated sperm (B pattern; progesterone vs. progesterone + depletion of calcium ion, 18 +/- 3.5% vs. 8 +/- 2.5%, p < 0.05, progesterone vs. progesterone + NiCl2, 20 +/- 3.8% vs. 6 +/- 5.2%, p < 01). Furthermore, genistein, a protein tyrosine phosphorylation inhibitor, inhibited progesterone-induced capacitation (B pattern; progesterone vs. progesterone + genistein, 20 +/- 3.8% vs. 11 +/- 2.4%, p < 01). In conclusion, progesterone induces capacitation in mouse sperm and this capacitation may be associated with calcium influx and tyrosine phosphorylation.

Animals↗

Sperm-immobilizing antibodies block capacitation in human spermatozoa.

Sperm-immobilizing antibodies block human fertilization by interfering with the acrosome reaction (AR). To clarify the mechanism of blockage of AR by sperm-immobilizing antibodies, the authors examined their effects on the increase of intracellular free Ca2+ concentration induced by follicular fluids (Ca2+ influx) in spermatozoa and on their capacitation. Sperm-immobilizing antibodies did not suppress Ca2+ influx induced by follicular fluid, but they inhibited capacitation of human spermatozoa. Namely delta%AR (%AR after addition of an AR inducer--%AR before treatment) induced by progesterone was significantly (p < .0001) lower when spermatozoa were incubated in human tubal fluid medium cotaining antibody-positive serum (1.2%), compared to that when incubated in control medium (19.2%). Furthermore, the proportion of both spermatozoa that became capacitated and ones that had become capacitated decreased significantly (p < .0001) after 2, 4, and 6 h of incubation in medium containing antisperm antibody-positive serum, compared to those of spermatozoa incubated in control medium. In conclusion, sperm-immobilizing antibodies may be closely related to their blockage of capacitation.

Acrosome Reaction↗

Concentration of granulocyte elastase in seminal plasma is not associated with sperm motility.

Although elastase could affect sperm motility in vitro, secretory leukocytes protease inhibitor (SLPI) prevents sperm from being attacked by elastase. The authors investigated the correlations of elastase level with sperm motility and SLPI level in vivo. Semen samples (n = 116) were collected and centrifuged after semen analysis. Elastase and SLPI levels were determined by an enzyme immunosorbent assay. Samples were classified by elastase levels into low (<250 ng/mL), moderate (250-1,000 ng/mL), and high elastase groups (> or =1,000 ng/mL). Elastase levels (range, 2.8-23,974.4 ng/mL) were not associated with sperm motility. The median SLPI level in the high elastase group was 15,900 ng/mL (range, 2.860-46,900 ng/mL). However, there was no significant correlation between elastase and SLPI levels in seminal plasma. Since SLPI forms a 1:1 complex with elastase, these results suggest that seminal plasma has a sufficient amount of SLPI to protect spermatozoa from elastase.

Enzyme Inhibitors↗