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T Yanaihara

Publications and source records attributed to T Yanaihara.

At least 19 recordsLinked to original sources
PubMed1998

Lactoferrin and interleukin-6 interaction in amniotic infection.

Lactoferrin (Lf) has been found in most biological fluids including amniotic fluid and cervical mucoids in pregnant women, and released from neutrophils in response to the inflammation. As Lf possesses antimicrobial properties, it is widely considered to be an important component of the host defence against microbial infections. It is known that premature labor is caused by amniotic infection with the increase of prostaglandin production. High concentration of the inflammatory cytokines: interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) in the amniotic fluid has been known. However, changes of Lf in amniotic fluid with infection has not been reported. In the present study, Lf concentrations in amniotic fluid were measured under the intra-uterine infections state and the biological significance of Lf was investigated. The effects of Lf on the IL-6 and IL-6mRNA production in cultured amnion cells were also investigated. The concentrations of Lf and IL-6 in amniotic fluid with CAM were 8.76 +/- 0.65 micrograms/ml and 6.92 +/- 4.88 ng/ml (n = 28) respectively and both were significantly higher (p < 0.01) than those without CAM [0.86 +/- 0.81 microgram/ml and 0.34 +/- 0.25 ng/ml (n = 31)]. Significant positive correlation (r = 0.91, p < 0.01) between Lf and IL-6 levels in amniotic fluid was found. IL-6 production induced by lipopolysaccharide (LPS) (100 ng/ml) in cultured amnion cells was significantly inhibited (p < 0.05) under the physiological concentration of Lf in amnion. Total RNA was extracted from the amniotic cells by guianizine solution. RT-PCR procedure and product analysis were performed from one microgram aliquote of total RNA. beta-actin was used as an international standard and c-DNA samples were followed by 30 cycles of PCR. RT-PCR product of IL-6 mRNA was detected by Southern hybridization. Expression of IL-6 mRNA was inhibited by the addition of Lf. From the results, the possibility that Lf might suppress amniotic IL-6 production under the condition of amniotic infection is suggested. It is also suggested that Lf might act as self defence mechanism from intra-uterine infection.

Amnion↗
PubMed1998

Analysis of HLA-DQ alpha sequences for prenatal diagnosis in single fetal cells from maternal blood.

We have extended a previously developed method that allows prenatal DNA diagnosis of female fetuses through the isolation of single nucleated erythrocytes from maternal blood by developing a method that can distinguish between maternal and fetal nucleated erythrocytes. Nucleated erythrocytes were separated by a density-gradient method and then collected by micromanipulation. Sex was determined after primer extension preamplification (PEP) of the entire genome of a single cell, and human leukocyte antigen (HLA)-DQ alpha type was determined after further amplification of this gene. The HLA-DQ alpha genotype of fetal erythrocytes in maternal blood samples and their corresponding paternal and maternal lymphocytes were successfully determined in all cases. The accuracy of the method was determined by using single nucleated erythrocytes from umbilical cord blood from five normal deliveries. This is the first demonstration that the fetal HLA-DQ alpha gene sequences can be identified in a small aliquot of a single nucleated erythrocyte in maternal blood. We believe that this method ushers in a new era in which the reliability and accuracy of noninvasive prenatal DNA diagnosis from maternal blood is markedly improved.

Alleles↗
PubMed1998

Prenatal diagnosis of ornithine transcarbamylase deficiency by using a single nucleated erythrocyte from maternal blood.

We have developed a method that allows the prenatal DNA diagnosis of ornithine transcarbamylase (OTC) deficiency by using a single fetal nucleated erythrocyte (NRBC) isolated from maternal blood. OTC gene analysis of a male patient (TF) with early onset OTC deficiency was performed by single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing. To investigate the possible prenatal diagnosis of OTC deficiency, maternal blood was obtained at 13 weeks of gestation of a subsequent pregnancy, from the mother of patient TF. NRBCs in the maternal blood were separated by using the density gradient method and then collected with a micromanipulator. The entire genome of a single NRBC was amplified by primer extension preamplification (PEP). The human leukocyte antigen (HLA)-DQ alpha genotype and sex were determined from small aliquots of the PEP product. The HLA-DQ alpha genotype of each of the parents of the male patient was also determined. Once a single NRBC had been identified as being of fetal origin, the OTC gene was analyzed by using the restriction fragment length polymorphism (RFLP) method. DNA analysis revealed a point mutation in exon 9 of the OTC gene in the OTC-deficient patient (TF). All NRBCs retrieved from maternal blood were successfully identified as being of fetal origin by HLA-DQ alpha genotyping and sex determination. RFLP analysis demonstrated that the fetal OTC gene was normal. This is the first study to successfully diagnose OTC deficiency prenatally, by using a single fetal NRBC from the maternal circulation. Such prenatal DNA diagnosis is non-invasive and can be applied to other genetic diseases, including autosomal and X-linked diseases.

Erythrocytes↗
PubMed1998

Inhibitory effects of catecholamines and maternal stress on aromatase activity in the fetal rat brain.

OBJECTIVE: Aromatization in the the fetal brain is thought to be involved both in sex differentiation during early development and in adult sexual behavior. Although recently the relationship between aromatase and catecholamine has been discussed, the effect of stress on aromatase in the fetal brain has not been clarified. Therefore, in the present study, localization of aromatase and the inhibitory effects of catecholamines and maternal stress on aromatase activity in the fetal rat brain were examined. METHODS: Localization of aromatase cytochrome P-450 using a specific polyclonal antiserum against human placenta aromatase was examined, and the inhibitory effects of dopamine and norepinephrine on aromatase activity in vitro were studied. Further, the influences of intrauterine stress on aromatase activity in the prenatal rat brain were evaluated in vivo. RESULTS: Aromatase-immunoreactive neurons are located principally in the medial amygdaloid nucleus. Aromatase activity in the fetal rat brain was competitively inhibited by dopamine and norepinephrine, with Ki values of 120 microM and 100 microM, respectively. Aromatase activity in the fetal brain was significantly lower in stressed rats given 1.5% salt water (89.2 +/- 17.5 fmol/mg/hr; n = 4) (p < 0.05) than in the control group (123.1 +/- 10.0 fmol/mg/hr; n = 4). CONCLUSION: Aromatase activity in the prenatal rat brain is influenced by catecholamine metabolism during intrauterine stress.

Amygdala↗
PubMed1997

Steroid sulfatase activity in osteoblast cells.

We have demonstrated steroid sulfatase activity in osteoblast cells and characteristics of the enzyme were also investigated. Cell free homogenate of rat osteoblast cell line, UMR106-01 and human osteoblast cell lines, MG-63, HOS were incubated with [3H] dehydroepiandrosterone-sulfate (DHEA-sulfate) or [3H] estrone-sulfate (E1-sulfate). The formation of DHEA or E1 from the corresponding substrate was identified by crystallization to constant specific activity. Michaelis constant (K(m)) for DHEA-sulfate was estimated as 2.1 x 10(-8)M in UMR106-01, 7.4 x 10(-7)M in MG-63, 5.8 x 10(-7)M in HOS and that for E1-sulfate was 4.1 x 10(-7)M, 3.0 x 10(-7)M, 9.8 x 10(-7)M, respectively. The expression of steroid sulfatase messenger ribonucleic acid in human osteoblast cells, HOS and MG-63 was first demonstrated by reverse transcription-polymerase chain reaction. The existence of steroid sulfatase in human and rat osteoblast cells suggests that osteoblast cells have the capacity to convert circulating sulfo-conjugated steroids to more active androgens and estrogens. This may indicate an important role of bone in facilitating hormonal action.

Animals↗
PubMed1997

A study of gamma-aminobutyric acid (GABA) in amniotic fluid.

OBJECTIVE: The purpose of the study was to evaluate the role of gamma-aminobutyric acid (GABA), an inhibitory neurotransmitter, in amniotic fluid (AF) during fetal distress, because it has been reported that several neurotransmitters, e.g. norepinephrine, are affected by GABA. METHODS: AF was obtained during elective cesarean section (CS, n = 11) and cesarean section due to fetal distress without labor pain (FD, n = 7). Maternal and umbilical-cord blood, as well as the first urine of the neonates, also were collected. GABA, norepinephrine (NE), and epinephrine (EP) concentrations were measured using HPLC. RESULTS: The GABA concentration was higher in the AF than in either maternal or fetal circulation, or in the first urine of neonates. The GABA concentrations in the AF and in the first urine of neonates were significantly higher in the FD group than in the CS group (p < 0.05). Furthermore, significant positive correlations were observed between the NE and GABA concentrations and between the EP and GABA concentrations in the AF. GABA was produced in a time-dependent manner in cultured amnion cells. CONCLUSION: The highest concentration of GABA was found in the AF. The GABA in the AF appeared to be derived from both the amniotic membrane and the fetal urine. The increase in the GABA concentration in cases of fetal distress might be partially derived from the fetus via fetal urine. The positive correlations between the concentrations of GABA and those of NE and EP in the AF, suggest that GABA, NE, and EP might play important roles during fetal distress.

Adrenergic alpha-Agonists↗
PubMed1997

3Beta-hydroxysteroid dehydrogenase activity in human osteoblast-like cells.

This paper describes for the first time the presence of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity in osteoblast-like cells and investigates its characteristics. 3beta-HSD activity was detected by the formation of androstenedione from [3H]dehydroepiandrosterone (DHEA) in whole cell assays of human osteoblast-like cells, HOS and MG-63. The radiolabeled product, androstenedione, was purified by thin-layer chromatography and identified by recrystallization on admixture with authentic androstenedione to show constant specific activities. The apparent Michaelis constant (Km) for DHEA in HOS was found to be 9.9 microM and that in MG-63 was 80.4 microM. The expression of the 3beta-HSD messenger ribonucleic acid in HOS and MG-63 was demonstrated through a reverse transcription-polymerase chain reaction. The PCR products were confirmed by Southern blot analysis. The existence of 3beta-HSD in osteoblast-like cells indicates that these cells convert delta5 androgens into more biologically active delta4 3-keto steroids. These results, together with the demonstration of other steroid converting enzyme systems, suggest that the osteoblast cells play an important role in facilitating hormonal action in bone tissue.

3-Hydroxysteroid Dehydrogenases↗
PubMed1997

[Gonadal dysfunction].

Function of hypothalamic-pituitary-ovarian axis is an essential factor for the maintenance of regular cycles in mature women. The disturbance of function of those organs causes gonadal dysfunction such as anovulation, amenorrhea and menstrual disorders. Therefore, the correct diagnosis for the assessment of CNS and ovarian function is clinically important to treat the patients those who have an menstrual disorders. In this review, the mechanism of normal gonadal cycles and the diagnostic method and the treatment of gonadal dysfunction are described.

Amenorrhea↗
PubMed1996

Behavioral and adrenocortical responses to stress in neonates and the stabilizing effects of maternal heartbeat on them.

Adrenocortical response to stress in neonates was assessed by estimating salivary concentrations of cortisol, dehydroepiandrosterone (DHEA) and dehydroepiandrosterone-sulfate (DHEA-S). Behavioral responses to stress were also scored clinically, according to the method of Lewis and Thomas [15]. The modulation of these measures by hearing maternal heartbeat was also evaluated. Levels of cortisol and DHEA in saliva in neonates correlated well with those of serum showing significant variation in painful stress. Hearing the maternal heartbeat significantly reduced the grade of variation, but the recorded sound of the Japanese drum with the same rhythm showed no significant effect. Scores of the behavioral response to stress also decreased significantly in neonates who were presented with the maternal heartbeat in comparison with those hearing no sound or the beat of a Japanese drum.

Circadian Rhythm↗
PubMed1996

Prenatal diagnosis of the fetal RhD blood type using a single fetal nucleated erythrocyte from maternal blood.

OBJECTIVE: To develop a method that allows prenatal diagnosis of the fetal RhD blood type from maternal blood. METHODS: Maternal blood was obtained at 8-31 weeks' gestation, and nucleated erythrocytes were separated with Percoll using a discontinuous density gradient method, then collected individually by micromanipulation under microscopic observation. After whole genome amplification with primer extension pre-amplification, exon 7 of the RhD and RhCE as well as the ZFX/ZFY loci were further amplified by a nested polymerase chain reaction (PCR). RESULTS: Nucleated erythrocytes were detected in nine of ten maternal blood samples, and sex was determined in 13 of 21 nucleated erythrocytes. RhD genotype could be diagnosed in 12 of the 13 nucleated erythrocytes in which sex could be determined. The results of RhD blood type and sex in nucleated erythrocytes obtained from maternal blood were identical with those of newborns. Fetal RhD blood type could be determined in six of ten maternal blood samples. CONCLUSION: A new method for noninvasive prenatal diagnosis of the fetal RhD blood type using a single nucleated erythrocyte isolated from maternal blood was demonstrated. This diagnostic method offers extremely useful information for the management of Rh-negative pregnant women. Furthermore, this method of prenatal diagnosis can be applied to other genetic disorders and is expected to become the preferred method of noninvasive prenatal diagnosis of DNA.

Base Sequence↗
PubMed1996

Osteoblast cells (MG-63 and HOS) have aromatase and 5 alpha-reductase activities.

Both aromatase and 5 alpha-reductase activities were found by whole-cell assay in osteoblast-like cells, MG-63 and HOS. Aromatase activity was measured by the [3H] water release method, and the formation of 5 alpha-androstanedione from androstenedione was expressed as 5 alpha-reductase activity. When CGS16949A, an inhibitor of aromatase, was added to the incubation medium at a concentration of 2 x 10(-9) M, sufficient to completely inhibit placental aromatase activity, only 63% to 68% inhibitions were observed. When progesterone, a competitive inhibitor of 5 alpha-reductase, was added at a concentration of 10(-5) M, 28% to 40% inhibitions were recorded. Because the release of [3H] from [1 beta-3H] androstenedione into water by 5 alpha-reductase is reported, results from the present study suggest that the measurement of aromatase activity in osteoblasts by the [2H] water release method may overestimate aromatase activity owing to the inclusion of 5 alpha-reductase activity. The results also suggest that osteoblast cells may play an important role in bone metabolism by transforming androgens into estrogens and more biologically active androgen derivatives.

Animals↗
PubMed1996

Aromatase and estrogen 2-hydroxylase activities of human placental microsomes in pregnancy-induced hypertension.

2-Hydroxylation is one of the major metabolic pathways of estrogens and is believed to be catalyzed by a form of cytochrome P450. Recently it has been reported that estrogen 2-hydroxylase activity in human placenta is catalyzed by aromatase. Some investigators suggested the effect of catechol estrogen on human placental steroidogenesis which may be related to pregnancy-induced hypertension (PIH) through the inhibition of catechol-O-methyltransferase (COMT) activity. In order to better understand the interrelationship between placental aromatase and estrogen 2-hydroxylase activities in PIH patients, both activities were evaluated in the PIH placentas. Human placental microsomes obtained from PIH patients were incubated with [1 beta-3H]androstenedione or [2-3H]estradiol in the presence of NADPH. Aromatase and estrogen 2-hydroxylase activities were assessed by the tritium water method. The immunosuppression patterns of both activities due to monoclonal antiaromatase cytochrome P450 antibody (MAb3-2C2) were studied. Estrogen 2-hydroxylase activity was significantly higher in PIH placentas (4.7 +/- 0.9 pmol/min/mg protein, n = 7) than in normal placentas (3.0 +/- 0.7 pmol/min/mg protein, n = 7). When the PIH placental microsomes were subjected to immunosuppression by 1 to 100 micrograms IgG of MAb3-2C2, estrogen 2-hydroxylase activity was suppressed by 94 to 65% whereas aromatase activity was strongly suppressed by 72 to 17%, respectively. From our results of high estrogen 2-hydroxylase activity in PIH placentas, it is assumed that there is a different estrogen catalyzing mechanism in PIH placentas.

Antibodies, Monoclonal↗
PubMed1996

A study of monoamine oxidase activity in fetal membranes.

OBJECTIVE: To study the role of decidual monoamine oxidase (MAO)-A and -B activities before delivery, the relationship between MAO activity in fetal membranes and catecholamine (CA) concentration in amniotic fluid (AF) was determined. METHODS: Fetal membranes and AF were obtained at the time of elective Cesarean section (CS group, n = 11) and Cesarean section due to fetal distress without labor pains (FD group, n = 5). MAO-A and -B activities were radiometrically measured using 14C-5-hydroxytriptamine for MAO-A substrate and 14C-benzylamine for MAO-B substrate. CA concentrations in AF were measured by high performance liquid chromatograph with an electro-chemical detector. RESULTS: Both MAO-A and -B activities in decidua obtained from CS were significantly lower than those obtained from FD. Both norepinephrine (NE) and epinephrine (EP) concentrations were significantly lower in the CS group than the FD group. A significant positive correlation between decidual MAO-A activity and NE concentration in AF was observed. No significant correlation was observed between MAO-B activity and the concentration of NE in AF. There was no correlation between EP concentrations and MAO activities. CONCLUSION: These results suggest that CA concentration in AF may be related to the activity of MAO in fetal membranes, determined by certain physiological processes during pregnancy. It has been suggested that metabolism of monoamines in fetal membranes also plays an important role in reducing monoamine influx into maternal myometrium from the AF.

Adult↗
PubMed1996

[Stage IIIa endometrial carcinoma: MR findings].

[PURPOSE]: An attempt was made to evaluate the ability of magnetic resonance (MR) imaging to diagnose stage IIIa endometrial carcinoma. [MATERIALS AND METHODS]: Thirty-three patients with endometrial carcinoma underwent MR imaging and surgery. Surgical staging was classified as I in 21 patients, II in 3 patients and III in 9 patients. The MR images of each patient were retrospectively reviewed by three radiologists. Only the clinical diagnosis of endometrial carcinoma was previously notified. Segmental disruption of the full thickness of the myometrium was considered serosal invasion. Intraperitoneal metastasis was diagnosed according to three criteria (intraperitoneal solid mass of isointensity compared with endometrial lesion, cystic mass excluding benign ovarian cysts, ascites). These evaluations were compared with the surgical findings and analyzed by the kappa statistic. [RESULTS]: The rates of sensitivity and positive predictive value (PPV) for serosal invasion were 33% and 6%, respectively. False positive evaluation frequently occurred when thickness of the intact myometrium was less than 5mm. The rates of sensitivity and PPV for intraperitoneal metastasis were 86% and 72%, respectively. The reason for false negative evaluation was small foci of intraperitoneal metastasis. Overall, sensitivity and PPV for stage IIIa were 86% and 69%, respectively. [CONCLUSION]: MRI was useful in detecting intraperitoneal metastasis of endometrial carcinoma with the exception of diagnosing serosal invasion. It is difficult to detect small foci of peritoneal metastasis. It is necessary to differentiate adnexal metastasis from benign adnexal masses.

Adenocarcinoma↗
PubMed1996

[Gynecological management of climacteric syndrome with psychiatric disorders].

PURPOSE: To discriminate patients with severe mental disorders from those complaining of menopausal symptoms, psychological tests were performed on patients who visited the menopausal clinic. The effectiveness of hormone replacement therapy (HRT) in patients with slight mental disorders was also evaluated. SUBJECTS AND METHODS: Patients with menopausal symptoms (n = 150, 41-59 yr. old) were interviewed by a psychiatrist (according to DSM-III-R) and classified as the climacteric syndrome group (C) and the mental disorder group (P). The patients were also evaluated by the Menopausal index (MI), Maudsley Personality Inventory (MPI) and Self-rating depression Scale (SDS). The patients with severe mental disorders were eliminated and the remaining patients with menopausal symptoms were treated with conjugated equine estrogen (0.625 mg/day) and medroxyprogesterone acetate (2.5mg/day) for 24 weeks. RESULTS: 1) Thirty six patients were diagnosed as P (24%) and seven of them were diagnosed as having major depression (4.7% of the all patients). 2) MPI-N (neurosis scale) in C was 16.2 +/- 10, and in P was 30.5 +/- 9.0, and SDS in C and P were 39.0 +/- 8.0, and 51.4 +/- 8.9, respectively. In both psychological test, P shows a significantly higher value than C (p < 0.01). 3) Ninety point six % of the patients responded. HRT was not effective in 63.2% of P, but when psychotropic drugs were combined, significant improvement (p < 0.05) was observed. CONCLUSION: One fourth of the patients were suspected of having mental disorders in the menopausal clinic, and 5% of the patients needed the care of the psychiatrist. And the application of MPI and SDS was useful in helping the gynecologist to differentiate and classify the mental disorders to some extent. The applicability and effectiveness of HRT for patients with mild mental disorders were suggested.

Adult↗
PubMed1996

[The effect of Shakuyaku-kanzo-to on prostaglandin production in human uterine myometrium].

Shakuyaku-kanzo-to (SK), a Chinese herb medicine consisting of Shakuyaku (Paeoniae Radix) and Kanzo (Glycyrrhizae Radix) has been used for the treatment of dysmenorrhea. It is reported that prostaglandin (PG) production increased during menstruation in uterine myometrium. To know the effect of SK on PG production in the uterine myometrium, the following in vitro study was undertaken. Human uterine myometrium was obtained from patients who underwent hysterectomy. 1. Myometrial cells were cultured and, SK, Shakuyaku (S), Kanzo (K), or glycyrrhetinic acid (GA), which is the major component of K, were added to the culture medium. Concentrations of PGE2, PGF2 alpha, and 6-ketoPGF1 alpha in the medium measured by RIA were significantly decreased by the addition of SK, K or GA but no effect was observed when S. 2. [3H]-arachidonic acid (AA) was added to culture medium and incorporated into the Sn-2 of phospholipids. [3H]-AA release and PG production in the medium were determined. PG were extracted and the radioactivity of PG was measured. The production of PGE2, PGE2 alpha and 6-ketoPGF1 alpha from labeled cells was significantly reduced by the addition of SK, K and GA. 3. Phosphatidylcholine containing [14C]-AA in Sn-2 (150,000 dpm) was incubated with cytosol of uterine myometrium and the amounts of [14C]-AA released were calculated as Phospholipase A2 activity. The amount of [14C]-AA release was inhibited dose-dependently by SK, K and GA. It is the first time that SK has been shown to suppress PG production in the myometrium by inhibiting cPLA2 activity.

Arachidonic Acid↗
PubMed1996

[Measurement of bone density by ultrasound bone densitometer in normal pregnant women].

The bone mineral density in reproductive aged and pregnant women was measured with an ultrasound bone densitometer (Achilles, Luner Co, USA). Both parameters of bone density on Os calsis, sound of speed (SOS) and broad ultrasound attenuation (BUA) were significantly correlated with bone mineral density in lumbar2-4 measured by dual energy X-ray absorptiometry (DXA) (p < 0.01) in reproductive aged women (mean age; 24.5 +/- 2.3, n = 30). The changes in bone density during pregnancy and at delivery were studied. In a cross-sectional study, no significant changes in SOS, BUA and stiffness were observed, but in a longitudinal study (n = 21), all of these parameters decreased as pregnancy advanced. The markers of bone turnover (serum Ca, alkaline phosphatase, procollagen type 1-C peptide, urinary Ca/Cr and deoxypyridinoline/Cr) did not change during pregnancy. Ten pregnant women with low bone density (less than-1.5 SD mean value) were found. There was no significant difference between the markers of bone turnover in pregnant women with low and normal bone density. The clinical usefulness of the screening of bone density of reproductive aged and pregnant women to detect osteopenia is suggested.

Adult↗