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Biomedical subjects

F Saji

Publications and source records attributed to F Saji.

At least 109 records · Page 6Linked to original sources
PubMed1992

Functional expression of human myometrial endothelin receptors in Xenopus laevis oocytes.

We demonstrate the existence of functional endothelin receptors in human uterine myometrium using the Xenopus oocyte expression system. Fifty nanograms of poly(A)+RNA from myometrium was injected into Xenopus laevis oocytes and incubated for 70-80 h. The membrane potential of the oocyte was clamped at -60 mV and membrane current was measured during and after endothelin stimulation. Endothelin-1 elicited a large inward membrane current in the oocytes injected with poly(A)+RNA; endothelin-2 elicited a small current; while endothelin-3 did not induce any membrane current. These results indicate the existence of messenger RNA encoding functional endothelin-1 receptors in human uterine myometrium.

Animals↗
PubMed1992

Preparation of oligozoospermic and/or asthenozoospermic semen for intrauterine insemination using the SpermPrep semen filtration column.

OBJECTIVE: To improve the quality of oligozoospermic and/or asthenozoospermic semen by the SpermPrep (Fertility Technologies Inc., Natick, MA) semen filtration column. DESIGN: The SpermPrep column was applied for semen manipulation in oligozoospermia and/or asthenozoospermia (sperm count less than 20 x 10(6)/mL, sperm motility less than 40%). After concentration of motile sperm using a 40% Percoll density gradient centrifugation, the sperm suspension was filtered through the SpermPrep column. The percentage yield of motile sperm by the SpermPrep method was compared with those by a two-layer Percoll density gradient (Pharmacia, Uppsala, Sweden) centrifugation and a swim-up method. Infertile couples with poor quality semen were treated with intrauterine insemination (IUI) with motile sperm by the three preparations through three cycles. SETTING: Department of Obstetrics and Gynecology, Osaka University Hospital. PATIENTS: Twenty-one couples with long-standing infertility because of poor quality semen. MAIN OUTCOME MEASURE: Recovery of motile sperm, sperm motility, and outcome of IUI were evaluated among three semen preparations. RESULTS: Motility was improved by the SpermPrep method in 32 of 33 cases of oligozoospermia and/or asthenozoospermia. Percentage yield of motile sperm by the SpermPrep method was significantly greater than those by the two-layer Percoll density gradient and swim-up methods (42.7 +/- 4.6 versus 22.1 +/- 3.1 and 13.8 +/- 3.5), but there is no significant difference in the sperm motility among three semen preparations. After one treatment cycle for each preparation, 2 of 21 women conceived after IUI with motile sperm separated in the SpermPrep method. CONCLUSIONS: The SpermPrep method is an improved semen manipulation method for oligozoospermia and/or asthenozoospermia.

Cell Separation↗
PubMed1992

Evaluation of acrosomal status using MH61-beads test and its clinical application.

OBJECTIVE: To evaluate the acrosomal status of viable sperm by MH61-beads test and its clinical application. DESIGN: Acrosomal status was evaluated using immunobeads coated with MH61 monoclonal antibody, which is specific for acrosome-reacted sperm. When viable sperm was coincubated with MH61-beads for an appropriate length of time, the formation of a sperm-bead complex was observed with a phase-contrast microscope, and the number of sperm binding to MH61-beads increased with the progression of the acrosome reaction. Using this agglutination, we developed the MH61-beads test to assess sperm function. SETTING: Department of Obstetrics and Gynecology, Osaka University Hospital. PARTICIPANTS: Forty-three volunteers and 20 males in our in vitro fertilization (IVF) and embryo transfer program. MAIN OUTCOME MEASUREMENT: The results of MH61-beads test were compared with the percentage of acrosome-reacted sperm detected by Pisium satinum agglutinin staining on 30 volunteers, with the results of sperm penetration assay (SPA) in 43 volunteers, and with the outcome of IVF in 20 patients. RESULTS: The results of MH61-beads test showed good reproducibility and correlated with the results of SPA using zona-free hamster eggs and IVF. CONCLUSIONS: The MH61-beads test, the results of which reflect the acrosomal status of sperm, may provide useful information concerning the fertilizing ability of sperm.

Acrosome↗
PubMed1991

The biological effects of macrophage-colony-stimulating factor induced by progestin on growth and differentiation of endometrial adenocarcinoma cells.

Progestins have biological effects of regression and differentiation on human endometrial adenocarcinoma. We investigated the effects of progestin on the induction of macrophage-colony-stimulating factor (M-CSF) and its receptor messenger RNAs in the human endometrial adenocarcinoma cell line Ishikawa which has receptors for both estrogen and progesterone. Poly(A)+RNA extracted from Ishikawa cells cultured with or without synthetic progestin R5020 was subjected to Northern blot hybridization using M-CSF and c-fms cDNA probes. The expression of M-CSF mRNA in Ishikawa cells increased about 2.3 times following treatment with R5020 at 10(-7) M. Induction of M-CSF mRNA by R5020 was antagonized by anti-progestin RU486 in a dose-dependent manner. However, c-fms mRNA, coding the M-CSF receptor, was expressed constitutively in Ishikawa cells and its expression was not affected by hormonal treatment. We further examined the biological effects of M-CSF on endometrial cancer cells. Colony formation of Ishikawa cells in soft agar, which represents anchorage-independent cell growth, was inhibited by M-CSF treatment. On the other hand, accumulation of glycogen granules in cytoplasm detected by periodicacid-Schiff staining was observed in Ishikawa cells treated with M-CSF. These results indicate that M-CSF, whose gene expression was enhanced by progestin, suppressed growth and induced differentiation of endometrial adenocarcinoma cells. These effects of M-CSF on endometrial cancer cells are similar to those of progestins, so the effects of progestins on these cells are, at least in part, probably mediated by M-CSF in an autocrine or paracrine manner.

Adenocarcinoma↗
PubMed1991

The enhanced production of placental interleukin-1 during labor and intrauterine infection.

The purpose of this study was to determine the effect of labor and chorioamnionitis in interleukin-1 production by human placenta. We studied the activity of the placenta to produce interleukin-1 with an enzyme immunoassay by culturing tissue blocks. The placental tissue obtained after labor produced a larger amount of interleukin-1 than placental tissue obtained before labor. All the placental tissues produced more interleukin-1 beta than interleukin-1 alpha. The placentas with labor and chorioamnionitis produced about seventeenfold more interleukin-1 than placentas with labor only. We immunohistochemically identified interleukin-1--producing cells in the placenta and found that syncytiotrophoblasts produced both interleukin-1 alpha and interleukin-1 beta, while Hofbauer cells produced only interleukin-1 beta. In vitro analysis of the trophoblast activities to produce interleukin-1 revealed that microbial byproducts enhanced interleukin-1 production, possibly inducing accumulation of interleukin-1 receptor-positive cells at the sites of inflammation. In addition to stimulation of prostaglandin biosynthesis and labor, the placental interleukin-1 may act as an inflammatory mediator, leading to systemic and local changes at fetomaternal interface and activating fetomaternal immune systems against intrauterine infection.

Chorioamnionitis↗
PubMed1991

Application of gene amplification by polymerase chain reaction to genetic analysis of molar mitochondrial DNA: the detection of anuclear empty ovum as the cause of complete mole.

To investigate the pathogenesis of complete hydatidiform mole (complete mole), we employed a newly developed gene amplification method by polymerase chain reaction (PCR) for the restriction fragment length polymorphism (RFLPs) analysis of extranuclear DNA (mitochondrial DNA) of complete mole. Whole cellular DNA was extracted from six molar tissues and from peripheral blood mononuclear cells of parents. Two hyperpolymorphic regions of mitochondrial DNA, a 1.5-kb-long fragment and a 1.9-kb-long fragment, were selectively amplified from the extracted DNA by the PCR method. The amplification products amounted to over 10 micrograms after 30 cycles of PCR. The PCR products were digested with endonucleases (HaeIII, HinfI, AluI, and TaqI) and then electrophoresed on agarose gel. The electrophoretic pattern of digested DNA showed that the RFLPs of molar mitochondrial DNA coincided with those of the patient, indicating that the mitochondrial DNA of complete mole was inherited from the ovum. As it has been identified that the intranuclear DNA of complete mole is transmitted only from the spermatozoa, our results verify that complete mole results from the fertilization of an anuclear "empty" ovum with normal sperm at the molecular level.

Base Sequence↗
PubMed1991

Analysis of immunoregulatory activity of a choriocarcinoma-derived factor: specific suppression of proliferative process of cell-mediated immune responses including LAK cell generation.

The immunosuppressive activity of a JEG-3 choriocarcinoma-derived factor in human IL-2-dependent T cell responses has been studied, together with its effect on IL-2-independent T cell responses induced by 10 nM TPA. The factor completely suppressed the IL-2-independent proliferative responses of T cells but failed to suppress antigenic expression of activation-associated CD 25 molecules. Further studies examined the effect of the factor on LAK cell generation induced by rIL-2. Recombinant IL-2-induced LAK cell proliferation was observed on Day 4 and Day 5, but not on Day 3. As the factor suppressed the responses of LAK cell proliferation, we tested whether it blocked the generation of Day 3, Day 4 and Day 5 LAK cells. The addition of the factor failed to suppress the generation of Day 3 LAK cells, while it partially suppressed the lytic activity of Day 4 LAK cells and completely suppressed that of Day 5 LAK cells. The data suggest the presence of a heterogeneous pattern for LAK cell generation; one without proliferation, but the other requiring proliferation, to acquire killer activity. Taken together with the evidence that the factor failed to suppress NK activity, the choriocarcinoma-derived factor suppressed only the proliferative events of immunocompetent cells, but inhibited neither their activation nor the differentiation events. This immunosuppressive factor might be involved in the prevention of host-mediated rejection of choriocarcinoma cells or maternal rejection of the fetus.

Choriocarcinoma↗
PubMed1991

Differential mRNA expression of three distinct classes of Fc gamma receptor at the feto-maternal interface.

Heterogeneous expression of three classes of Fc gamma receptor (Fc gamma RI, IIa, IIb, and III) in the human placenta and decidua was examined by Northern blot hybridization and cDNA amplification analysis by polymerase chain reaction. Messenger RNA of Fc gamma RI, IIa and III genes were consistently expressed in the human placenta in all trimesters of gestation. The transcripts of the Fc gamma RIIb gene, on the other hand, dramatically increased in placentae at the second and third trimesters. This characteristic expression of Fc gamma RIIb after 20 gestational weeks was confirmed by sequential cDNA amplification analysis. Fc gamma RI, IIa and III mRNAs, but not Fc gamma RIIb, were also detected in the human decidua. Interestingly, while Fc gamma R mRNA could be induced in uterine endometrium by pseudopregnancy therapy using estrogen and progesterone, there was no detectable mRNA in hormone-unprimed normal endometrium. These findings suggest that Fc gamma Rs expressed at the feto-maternal interface can be transcriptionally regulated by sex steroid hormones as multifunctional molecules. In addition, the Fc gamma RIIb molecule is predominantly produced by placental tissues after the mid-trimester of gestation and possibly plays an important role in the transport of IgG molecules from mother to fetus.

Antigens, CD↗
PubMed1991

The gene expressions of macrophage colony-stimulating factor (MCSF) and MCSF receptor in the human myometrium during pregnancy: regulation by sex steroid hormones.

We investigated the biological effect of sex-steroid hormones, secreted from the corpus luteum and placenta, on the induction of mRNA encoding macrophage colony-stimulating factor (MCSF) and c-fms proto-oncogene (MCSF receptor) in the human uterine myometrium. Poly(A)+RNA was extracted from the myometrium of pregnant and non-pregnant uterine myometrium and then Northern blot analysis was performed on poly(A)+RNA. The myometrium of non-pregnant women expressed neither mRNA of macrophage colony-stimulating factor (MCSF) nor any transcript related to the c-fms proto-oncogene. On the other hand the myometrium of pregnant women expressed MCSF mRNA (4.7 kb) and two kinds of transcript related to the c-fms proto-oncogene (3.9 and 1.3 kb). The mRNAs of both MCSF and c-fms proto-oncogene were induced in the uterine myometrium of non-pregnant women under pseudopregnant therapy of mestranol and norethindrone. These results indicate that sex steroid hormone secreted from the corpus luteum of pregnancy and/or placenta may be deeply involved in the hypertrophic change of uterus during pregnancy by inducing MCSF and MCSF receptor (c-fms proto-oncogene protein product) in the myometrium.

Corpus Luteum↗
PubMed1991

Probabilistic assessment of the HLA sharing of recurrent spontaneous abortion couples in the Japanese population.

In spite of a number of investigations, the concept of human leukocyte antigen (HLA) sharing in recurrent spontaneous abortion (RSA) couples remains controversial. We introduced the basal antigen sharing rate (BSR) by the original mathematical approach using the gene frequency of all HLA specificities derived from our regional control population and applied this parameter for the comparison with RSA couples. RSA couples were classified into three subgroups; primary (3 or more consecutive abortions), secondary (3 or more consecutive abortions after 1 live birth), and potential (2 consecutive abortions) aborters. No significant differences between the HLA class I sharing rates (one or more antigens shared on a single locus) of the all RSA subgroups and the calculated BSRs were observed. In the HLA-DR and DQ loci, on the other hand, the antigen sharing rates of primary aborters were significantly higher than BSRs (p less than 0.01/DR, p less than 0.05/DQ). While potential aborters showed a result similar to that of the primary aborters, no significant antigen sharing of HLA class II was observed in secondary aborters. Our data suggest that BSR is a useful parameter for detection of significant HLA sharing in regional populations and the consecutive abortions that occurred primarily are certainly relevant to HLA class II sharing.

Abortion, Spontaneous↗
PubMed1991

Trophoblast-derived interleukin-1 (IL-1) stimulates the release of human chorionic gonadotropin by activating IL-6 and IL-6-receptor system in first trimester human trophoblasts.

Interleukin-1 (IL-1) has a unique activity to stimulate the release of multiple hormones in a number of human and murine endocrine systems. IL-6 also expresses such activities by activating IL-6-receptor (R)-mediated signal transduction pathways. Since the placenta produces both of these cytokines and endocrine hormones such as hCG, we investigated how these cytokines regulate hCG release by normal trophoblasts. Trophoblasts purified by Percoll density gradient released hCG from 120 min after stimulation with recombinant (r) IL-1 alpha, and its release was dependent on the rIL-1 alpha concentration used. The rIL-1 alpha-stimulated trophoblasts released a molecule with IL-6 activity antecedently, as determined by an IL-6-dependent cell line, MH60.BSF2 cells. The IL-6 identity of the released molecule was confirmed by goat anti-IL-6 antiserum. rIL-1-mediated hCG release from trophoblasts was completely abrogated to the basal level by pretreatment of the trophoblasts with PM1, an anti-IL-6-R monoclonal antibody. Identical results were observed with rIL-1 beta. These results showed that rIL-1-induced hCG release was totally dependent on IL-6- and IL-6-R-mediated signal transduction in human trophoblasts. The presence of peripheral monocytes in the purified trophoblast fraction, however, induced a rapid decrease in IL-6 and hCG release after their maximal release, suggesting some regulatory interaction between trophoblasts and the monocytes. In contrast, rIL-1-mediated enhancement of IL-6 and hCG secretion by purified trophoblasts was no longer observed at 24 h compared with that of the unstimulated trophoblasts, while spontaneous hCG secretion was significantly inhibited by pretreatment of trophoblasts with PM1. The results showed that IL-6 and hCG secretion might also be regulated by a number of agents besides IL-1, and that hCG secretion as well as its release is dependent on IL-6 and IL-6-R system in trophoblasts.

Antibodies↗
PubMed1990

Perinatal infection of human T-lymphotropic virus type I, the etiologic virus of adult T-cell leukemia/lymphoma. DNA amplification of specific human T-lymphotropic virus type I sequences.

A gene amplification technique, polymerase chain reaction, was used to detect human T-lymphotropic virus type I (HTLV-I), the etiologic agent of adult T-cell leukemia/lymphoma, in mononuclear cells in peripheral blood and breast milk of ten HTLV-I carrier gravida. The DNA in umbilical cord blood mononuclear cells of the neonates born to the HTLV-I carrier gravida were also amplified and examined for the possibility of HTLV-I infection via placenta during pregnancy. The HTLV-I sequences were detected both in the peripheral blood and milk of all ten carrier gravida by Southern blot analysis of amplified DNA. However, HTLV-I proviral DNA could not be detected in the cord blood of the carriers' neonates, indicating that transplacental infection of HTLV-I should be rare and that postpartum infection via breast milk is a likely major perinatal transmission route.

Antibodies, Neoplasm↗
PubMed1990

Studies on the pathogenesis of choriocarcinoma by analysis of restriction fragment length polymorphisms.

The association of complete hydatidiform mole with choriocarcinoma has long been recognized, but it is unknown whether the pathogenesis of the two are identical. We investigated the pathogenesis of these trophoblastic tumors by analyzing restriction fragment length polymorphisms using a minisatellite DNA probe to choriocarcinoma, the complete mole, and normal trophoblasts as well as the parental cells. The polymorphic fragments of the complete mole were all transmitted from the paternal DNA, but some polymorphic fragments of the paternal DNA were not recognized in the complete mole. This confirms at a molecular level the androgenetic origin of the complete mole. In some cases of choriocarcinoma, the pattern of inheritance of restriction fragment length polymorphisms was the same as that in the complete mole, whereas in others all the polymorphic fragments in tumor tissues were identical to those in the host DNA. These results suggest that the pathogenesis of choriocarcinoma varies, being completely different from that of the complete hydatidiform mole in some cases.

Choriocarcinoma↗
PubMed1990

Differential diagnosis between complete mole and hydropic abortus by deoxyribonucleic acid fingerprints.

We used a new method of deoxyribonucleic acid fingerprint analysis to obtain the differential diagnosis between complete mole and hydropic abortus. This method with a deoxyribonucleic acid minisatellite probe requires only a small amount of tissue sample and peripheral blood, and presents individual specific restriction fragment length polymorphisms (deoxyribonucleic acid "fingerprints") by simultaneous detection of many hypervariable regions (minisatellite regions) widely dispersed in the human genome. Southern blot hybridization showed that in cases of complete mole, all polymorphic fragments were exclusively inherited from the father. Some of the polymorphic bands of paternal deoxyribonucleic acid were not observed in molar deoxyribonucleic acid. However, in the hydropic abortus, the polymorphic fragments could be traced back to its parent. These results indicate that deoxyribonucleic acid fingerprints could distinguish the abnormal fertilization of complete mole (androgenesis) from the normal fertilization of hydropic abortus by identifying the difference in genetic variations between complete mole and hydropic abortus at the deoxyribonucleic acid level.

Blotting, Southern↗
PubMed1990

In vitro and in vivo production of interleukin-6 by fetal mononuclear cells.

We examined the functional activity of cord mononuclear cells (MNCs) to produce interleukin (IL)-6 in vitro and in vivo. We stimulated fetal T-cell, B-cell, and macrophage fractions with mitogens. The supernatant of each stimulated cord cell fraction contained a comparable amount of IL-6 to that of each adult cell fraction activated similarly, suggesting functional maturity of cord MNCs' ability to produce IL-6. We then examined fetal cells' activities to produce IL-6 in response to perinatal infections, especially to intraamniotic infections (IAI). Among the cord MNCs from fetuses with IAI, macrophages were major cells producing IL-6. The serum IL-6 level in the fetuses with IAI was elevated, but it decreased to normal levels after antibiotic treatment; this finding indicates that IL-6-mediated host defense mechanisms by cord MNCs are triggered by perinatal infections.

Amniotic Fluid↗
PubMed1990

Effect of alcohol sulfate, linear alkylbenzene sulfonate and natural soap on the development of fertilized eggs of the mouse in vitro.

Eggs from B6 x C3F1 female mice, which were fertilized in vitro with sperm from C3 x 101F1 male mice, were treated with synthetic surfactants, alcohol sulfate (AS) and linear alkylbenzene sulfonate (LAS), and natural soap for 1 h at the pronucleus stage, and then cultivated for 5 days. Eggs treated with AS or LAS at concentrations of less than 0.025% developed to the blastocyst stage as well as the untreated ones. At concentrations of AS or LAS higher than 0.03% no egg developed beyond the 1-cell stage. There appeared to be a threshold concentration between 0.025% and 0.03% of AS or LAS on the development of the mouse egg. However, natural soap had no effect on the development of the mouse egg up to 0.05%. When AS or LAS was applied to the culture medium throughout the cultivation of fertilized eggs for 5 days, there also appeared to be a threshold concentration between 0.01% and 0.025%, but not in the case of natural soap. The results provide additional support to our previous observations that AS and LAS can interrupt mouse pregnancy by killing fertilized eggs.

Alcohols↗
PubMed1990

Expression of the corticotropin-releasing hormone (CRH) gene in human placenta and amniotic membrane.

Immunoreactive corticotropin-releasing hormone (IR-CRH) in maternal plasma increases progressively during pregnancy and decreases rapidly after delivery, suggesting that IR-CRH is produced in the placenta. We studied the expression of the CRH gene in developing human chorionic tissue, the amniotic membrane, the uterine myometrium and a fresh surgical specimen of hydatidiform mole by Northern blot analysis. Our results were as follows: (1) CRH mRNA was demonstrated in the placenta in the third trimester and at term, but under detectable level in the first and second trimesters. (2) CRH mRNA expression was observed in the amniotic membrane, but its expression in the myometrium in normal pregnancy was under detectable level at term. (3) CRH mRNA was also under detectable level in trophoblasts of a hydatidiform mole. These results suggest that the sources of the increased level of IR-CRH in human plasma and amniotic fluid during pregnancy are the placenta and amniotic membrane, and that gene expression of placental CRH increases during pregnancy.

Amnion↗