[The structural and functional characterization of P-450.aromatase (estrogen synthetase)].
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Amino Acid Sequence↗
Biomedical subjects
N Harada
Publications and source records attributed to N Harada.
[Two cases of blind pouch syndrome].
Case 1. A 39-yr-old male was presented with anemia. Initial blood examination showed remarkable iron deficiency anemia. Small bowel X-ray and abdominal CT examination revealed a dilated tract at the end of the ileum. Surgical operation was done. The surgical specimen showed side to side anastomosis which was reconstructed at the operation for rt. inguinal herniation 46 days after his birth. The oral side of the tract was remarkably dilated. Case 2. A 50-yr-old male was presented with dizziness. He had anemia and clubbed fingers. A small bowel X-ray examination was done and it showed side to side anastomosis in the middle of the small intestine. Surgical operation was performed. The surgical specimen showed side to side anastomosis about 2 m 20 cm from the Treitz' ligament, which was reconstructed during the operation for ileus when he was 10 years old. The blind portion was remarkably dilated. In both case, the anemia was improved after the operation.
Adult↗
Genetic studies to characterize the origin of the mutation in placental aromatase deficiency.
Placental aromatase deficiency has recently been shown to be due to expression of RNA transcripts encoding abnormal aromatase molecules with 29 extra amino acids. To establish whether this aromatase deficiency is a hereditary or sporadic disease, we examined the genetic defect of the aromatase gene in the family of a patient. Direct sequencing of fragments of the aromatase gene prepared by PCR revealed that the splicing donor sequence (GT) of intron 6 in controls was mutated to GC in the patient, whereas the parents showed signals of both GT and GC. Subcloning of PCR products of the parents gave two different types of clones with GT and GC sequences in this site. Furthermore, for diagnosis of this deficiency, competitive-oligo-nucleotide-priming PCR of genomic DNA was performed in the presence of both normal and mutational oligonucleotide primers labeled with two kinds of fluorescent dyes, and the products were separated by agarose gel electrophoresis and were detected fluorometrically in the gel. Genomic DNA of the patient gave a PCR product primed only by the mutational primer, whereas that of controls gave a product primed only by the normal primer. The PCR products of the parents were primed by both primers. The results obtained by this fluorometric method were also confirmed by differential hybridizations with specific oligonucleotide probes. Thus these findings indicate that this deficiency is an autosomal hereditary disease and that the patient is a homozygote, while the parents are heterozygotes, for this mutation.
Aromatase↗
[A case of heterotopic pancreas in the ileum].
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Adult↗
Molecular study of the Prader-Willi syndrome: deletion, RFLP, and phenotype analyses of 50 patients.
Deletion and RFLP studies with 5 cloned DNA markers localized at 15q11.2 were performed in 50 patients with the Prader-Willi syndrome (PWS). A one-copy density (deletion) for at least one of 4 loci, D15S9, D15S11, D15S10, D15S12, was detected in 32 (64%) of the 50 patients; deletions of each of the 4 loci were found in 29, 30, 29, and 28 patients, respectively. Three patients showed 4 or more copy density for D15S12 locus, in addition to deletions. The remaining 18 patients showed two-copy densities for each of the 4 loci. A common site of rearrangements among our 32 patients as well as the reported patients seemed to be confined to a segment between D15S9 and D15S11, suggesting the putative PWS gene locus in this segment. Of 6 patients who have cytologic deletions but did not show any molecular deletions, 3 have normal size of hands and feet, and 4 have normally pigmented skin and hair. The normal pigmentation was also observed in 3 patients who had small molecular deletions in the examined 5-locus segment. These observations may support the conception of contiguous gene syndrome. RFLP analysis demonstrated maternal uniparental isodisomy of chromosomes 15 in both a patient with 45,t(15q;15q) and a karyotypically normal patient. Based on the results of the present study, a new model is proposed to explain the occurrence of PWS with a variety of chromosome abnormalities, including partial monosomy, disomy, trisomy, and/or tetrasomy for 15q11.2. The normal development may require an even or more "number ratio" of paternally derived allele(s) to maternally derived allele(s) of the gene(s) localized at 15q11.2, and a disturbance of the ratio would lead to the PWS phenotype.
Adolescent↗
Neuroanatomical specificity in the co-localization of aromatase and estrogen receptors.
The relative distributions of aromatase and of estrogen receptors were studied in the brain of the Japanese quail by a double-label immunocytochemical technique. Aromatase immunoreactive cells (ARO-ir) were found in the medial preoptic nucleus, in the septal region, and in a large cell cluster extending from the dorso-lateral aspect of the ventromedial nucleus of the hypothalamus to the tuber at the level of the nucleus inferioris hypothalami. Immunoreactive estrogen receptors (ER) were also found in each of these brain areas but their distribution was much broader and included larger parts of the preoptic, septal, and tuberal regions. In the ventromedial and tuberal hypothalamus, the majority of the ARO-ir cells (over 75%) also contained immunoreactive ER. By contrast, very few of the ARO-ir cells were double-labeled in the preoptic area and in the septum. More than 80% of the aromatase-containing cells contained no ER in these regions. This suggests that the estrogens, which are formed centrally by aromatization of testosterone, might not exert their biological effects through binding with the classical nuclear ER. The fact that significant amounts of aromatase activity are found in synaptosomes purified by differential centrifugation and that aromatase immunoreactivity is observed at the electron microscope level in synaptic boutons suggests that aromatase might produce estrogens that act at the synaptic level as neurohormones or neuromodulators.
Animals↗
Possible mapping of the gene for transient myeloproliferative syndrome at 21q11.2.
The parental origin of the extra chromosome 21 was studied in 20 patients with trisomy 21-associated transient myeloproliferative syndrome (TMS) using chromosomal heteromorphisms as markers; this was combined with a study of DNA polymorphisms in 5 patients. Of these, 10 were shown to result from duplication of a parental chromosome 21, viz., maternal in 8 and paternal in 2. A patient with Down syndrome-associated TMS had a paracentric inversion in two of his three chromosomes 21 [47,XY,-21,+inv(21)(q11.2q22.13)mat,+inv(21)(q11.2 q22.13)mat]. These findings support our hypothesis of "disomic homozygosity" of a mutant gene on chromosome 21 in 21-trisomic cells as being a mechanism responsible for the occurrence of TMS. The finding also suggests that the putative TMS gene locus is at either 21q11.2 or 21q22.13, assuming that the gene is interrupted at either site because of the inversion. The study of 5 TMS patients using DNA polymorphic markers detected a cross-over site on the duplicated chromosomes 21 between 21q11.2 (or q21.2) and 21q21.3 in one patient, and a site between 21q21.3 and q22.3 in another patient, evidence that confined the gene locus to the 21cen-q21.3 segment. These findings suggest that the putative TMS gene is located at 21q11.2. The extra chromosome 21 in the latter two TMS patients probably resulted from maternal second meiotic non-disjunction, in view of the presence of recombinant heterozygous segments on their duplicated chromosomes 21.
Chromosome Aberrations↗
Distribution of aromatase-immunoreactive cells in the mouse forebrain.
The distribution of aromatase-immunoreactive cells was studied by immunocytochemistry in the mouse forebrain using a purified polyclonal antibody raised against human placental aromatase. Labeled perikarya were found in the dorso-lateral parts of the medial and tuberal hypothalamus. Positive cells filled an area extending between the subincertal nucleus in the dorsal part, the ventromedial hypothalamic nucleus in the ventral part, and the internal capsule and the magnocellular nucleus of the lateral hypothalamus in the lateral part. The same distribution was seen in the two strains of mice that were studied (Jackson and Swiss), and the number of immunoreactive perikarya did not seem to be affected by castration or testosterone treatment. No immunoreactivity could be detected in the medial regions of the preoptic area and hypothalamus; these were expected to contain the enzyme based on assays of aromatase activity performed in rats and on indirect autoradiographic evidence in mice. Our data raise questions concerning the distribution of aromatase in the brain and the mode of action of the centrally produced estrogens.
Animals↗
Evaluation of murine interleukin 4 (IL-4) receptor expression using anti-receptor monoclonal antibodies and S1 nuclease protection analyses.
Anti-receptor antibodies have previously been used in two cytokine systems (IL-1 and TNF alpha) to identify the existence of different cytokine receptors on different cell types. In this study, we have similarly used two approaches to evaluate whether IL-4 receptors on different cell types are identical, or whether more than one species of IL-4 receptor exists. The first approach involved production of monoclonal antibodies specific for the IL-4 receptor expressed by the murine mast cell line, MC/9. Six anti-IL-4 receptor monoclonal antibodies were produced against the purified soluble extracellular domain of the recombinant IL-4 receptor derived from MC/9 cells. These antibodies were capable of binding to and specifically immunoprecipitating the soluble extracellular domain of the recombinant mast cell IL-4 receptor. Following biotinylation of the antibodies and addition of phycoerythrin-streptavidin, their binding to cell associated IL-4 receptors on MC/9 mast cells could be readily visualized by immunofluorescence. Using this approach, the anti-mast cell IL-4R antibodies were found to specifically bind IL-4 receptors expressed on a variety of other murine cell types, including T cells, B cells, macrophages, fibroblasts, and L cells. The antibodies did not bind to two human cell lines known to bind human but not murine IL-4. The intensity of staining was directly related to the number of IL-4 binding sites identified previously by receptor-ligand equilibrium binding analyses. As a second approach to evaluating potential receptor heterogeneity, we constructed S1 nuclease protection assay probes for two separate regions of the mast cell IL-4 receptor, one located in the extracellular domain and one in the intracellular domain. Subsequent S1 analyses showed that both regions are expressed by the following types of cells: T cells, B cells, macrophages, myeloid cells, L cells, and stromal cells. The two approaches used in this study therefore indicate that the same or highly similar IL-4 receptor species is expressed by a wide variety of hemopoietic and nonhemopoietic cells. Since the anti-IL-4 receptor antibodies produced in this study did not block binding of IL-4 to its receptor, we cannot exclude the possible existence of a second type of IL-4R coexpressed on the cells tested in this study, or expressed uniquely by other cell types that were not investigated.
Animals↗
The direct contractile effect of gastrin releasing peptide on isolated gastric smooth muscle cells of the guinea pig.
Smooth muscle cells isolated from the gastric muscle layers of the guinea pig were used to determine whether gastrin releasing peptide (GRP) can cause contraction by exerting a direct action on muscle cells. In addition, the inhibitory effect of 8-( N,N-diethylamino )-octyl-3,4,5-trimethoxybenzoate hydrochloride ( TMB-8 ), an inhibitor of intracellular Ca2+ release, and verapamil, a Ca2+ channel blocker, on the GRP-induced contraction of gastric smooth muscle cells were examined. GRP elicited a contractile response of gastric muscle cells in a dose-dependent manner. The ED50 was 13 pM. TMB-8 significantly inhibited the contractile effect of GRP in gastric muscle cells. These results demonstrate the direct action of GRP on the gastric smooth muscle cells of the guinea pig, and the importance of Ca(2+)-release from intracellular calcium, stores in the contractile response to GRP.
Animals↗
Prevalence of Raynaud's phenomenon in Japanese males and females.
In order to estimate the prevalence rate of Raynaud's phenomenon in the Japanese population, 1875 males and 1998 females were interviewed by medical doctors, aided by a typical photograph of Raynaud's phenomenon. The prevalence rates were 3.3% in males and 2.5% in females. Approximately 60% of the male cases of Raynaud's phenomenon were presumed to be caused by vibration syndrome or trauma to the fingers. Prevalence rates excluding these two causes were 1.2% in males and 2.2% in females. The male:female ratio of these prevalence rates was 1:1.9. The prevalence rate for males increased with age, while that for females tended to decrease. The male:female ratio for subjects under 50 years old was 1:5.2 and for those 50 years old or older the ratio was 1:1.1. The prevalence rates of symptoms limited to finger whitening, excluding cases caused by vibration syndrome or trauma, were 1.0% in males and 1.8% in females. Both of these prevalence rates were lower than those of male and female Caucasians.
Adult↗
Calcium distribution and mobilization in single rat parotid acinar cells investigated by digital imaging microscopy.
The intracellular distribution and mobilization of cytosolic free calcium in single rat parotid acinar cells was analyzed by a digital imaging microscope equipped with a microspectrofluorometer, using calcium-sensitive dye fura-2. In the resting state, intracellular distribution of cytosolic free calcium concentration ([Ca2+]i) was heterogeneous: [Ca2+]i in the nuclear and perinuclear region was usually higher than that in the cytoplasm. By Ca(2+)-ionophore ionomycin and muscarinic agonist carbachol stimulation in the presence of 1 mM extracellular Ca2+, [Ca2+]i increased markedly and the gradient of [Ca2+]i between the nuclear region and the cytoplasm decreased. In ionomycin stimulation, [Ca2+]i increased homogeneously and this homogeneous increase was irreversible. In carbachol stimulation the gradient of [Ca2+]i between the nuclear region and the cytoplasm obviously reappeared within 2 min. By carbachol stimulation in the absence of extracellular Ca2+ (added 1 mM EGTA), [Ca2+]i returned to the prestimulation level after the initial transient increase. The distribution of [Ca2+]i also returned approximately to the prestimulation state. The gradient of [Ca2+]i between the nuclear region and the cytoplasm did not disappear even when [Ca2+]i elevated at the peak value.
Animals↗
XY translocation in a boy with ichthyosis, hypogonadism, short stature and mental retardation.
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Abnormalities, Multiple↗
De novo interstitial deletion of 1p (pter----p34.1::p32.3----qter).
We report a case of a 9 month old girl with a de novo interstitial deletion of 1p, karyotype 46,XX, del(1)(pter----p34.1::p32.3----qter). She had dysmorphic features including upward slanting palpebral fissures, a bulbous nose, a long philtrum, low set and malformed ears, a short neck, hypoplastic nails on both index fingers, widened interdigital spaces between the toes, dilated lateral ventricles, right hydronephrosis, a dilated right ureter, mental and motor developmental delay, and generalised hypotonia.
Abnormalities, Multiple↗
Factors influencing vibration sense thresholds used to assess occupational exposures to hand transmitted vibration.
The effects of various conditions, including temporary threshold shifts (TTS) induced by exposure to vibration on vibration sense thresholds, have been investigated. The vibration sense thresholds of five subjects were measured on the middle fingertip of the left hand. A contactor with a diameter of 7 mm was surrounded by three alternative plates with holes of different sizes. The contact force was controlled at either 1 N, 2 N, or 3 N. For the TTS test, the left hand was exposed to vibration at 20 ms-2 rms for five minutes. The frequencies of both the exposure to vibration and the vibration threshold test were in the range 16 Hz to 500 Hz. Using a surround around the contactor greatly reduced the vibration sense threshold at 16 Hz and 31.5 Hz but increased the threshold at 125 Hz, 250 Hz, and 500 Hz. An effect of contact force was seen only at the higher frequencies; larger contact forces led to lower thresholds at 125 Hz, 250 Hz, and 500 Hz. As temperature of the finger skin decreased, the vibration thresholds increased, with the changes at higher frequencies greater than those at lower frequencies. The TTS at 16 Hz and 31.5 Hz measured 0.5 minutes after exposure to vibration (TTS0.5) were highest after exposures to vibration at lower frequencies. The TTS0.5 at 63 Hz was similar after exposure to all frequencies. The TTS0.5 values at 125 Hz, 250 Hz, and 500 Hz were highest after exposures to vibration at 125 Hz and 250 Hz. It was apparent that the physiological characteristics of vibration sensation at low and high frequencies differed significantly. These findings suggest that two representative frequencies can be used when evaluation the neurological effects of occupational exposures to vibration by means of vibration sense thresholds.
Adult↗
Isolation of and calcium mobilization in vestibular hair cells of the guinea pig.
Vestibular hair cells were isolated from the macula utriculi, macula sacculi and crista ampullaris of the guinea pig, using enzymatic and mechanical techniques. The cells could be classified into two types: flask-shaped ones with a neck and rod- or round-shaped ones without a neck. The flask-shaped cells were thought to be type I cells, the rod-shaped cells type II cells, while the round-shaped cells could have been type I or type II cells. The intracellular free calcium ion concentrations [( Ca2+]i) of these cells were determined using the Ca2(+)-sensitive dye Fura-2 and digital imaging miroscopy. In the resting state, [Ca2+]i in the nucleus and cuticular plate was variable in both types of cells. The 150 mM KCl stimulation, which might induce a depolarization, resulted in a temporary increase in [Ca2+]i in both types of cells. In the presence of 1 micron ionomycin, there was an irreversible increase in [Ca2+]i in both types of cells. These observations aid in elucidating vestibular function.
Animals↗
[Mechanism of protective effects of ouren-gedoku-to and san'ou-syashin-to on the gastric mucosa].
Ouren-Gedoku-To (OGT) and San'ou-Syashin-To (SST), traditional Chinese prescriptions, have been used to treat peptic ulcer. In order to elucidate the mechanism of the protective effects of OGT and SST on the gastric mucosa, we studied their effects on gastric mucosal lesions, ion permeability and prostaglandin synthesis using ulcer experimentally induced in rats by saturation with acidic test solution after taurocholate treatment. Both OGT and SST inhibited taurocholate-induced gastric mucosal lesions in a dose-related manner. These prescriptions dose-dependently inhibited not only the increase of hydrogen and sodium ion net fluxes but also possible hydrogen ion back diffusion into the gastric mucosa during saturation with acidic test solution. SST induced a significant increase in gastric mucosal prostaglandins synthesized from 14C labeled arachidonate. These results suggest that both OGT and SST by strengthening the gastric mucosal barrier demonstrate their protective effects on the gastric mucosa and that this effect of SST is attributable to the increased ability of the gastric mucosa to synthesize prostaglandin.
Animals↗
Isolation of and calcium kinetics in cochlear inner hair cells of the guinea pig.
Single inner hair cells of the guinea pig cochlea were isolated using enzymatic and mechanical techniques. The intracellular free calcium ion concentrations [( Ca2+]i) of the isolated inner hair cells were determined using the Ca2+ sensitive dye fura-2 and digital imaging microscopy. In the presence of 1 micron ionomycin, a Ca2+ ionophore, there was an irreversible increase in [Ca2+]i. The 150 mM KCl stimulation, which induces a depolarization, resulted in a temporary increase in [Ca2+]i. This increase in [Ca2+]i was not observed under conditions of depolarization, in Ca(2+)-free medium. These observations are interpreted to mean that the [Ca2+]i during membrane depolarization mainly originates from an influx of extracellular Ca2+ into the cytoplasm.
Animals↗