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Biomedical subjects

H Okamura

Publications and source records attributed to H Okamura.

At least 541 records · Page 30Linked to original sources

[Long-term follow-up of tetralogy of Fallot corrected by Okamura's method under simple deep hypothermia].

Corrective surgeries for TOF (Tetralogy of Fallot) were performed in 350 patients from 1967 to 1983 by Okamura's method under simple deep hypothermia. Radical operations were done in all patients, including very young children. The operative techniques had several differences from the standard procedure. Patients were operated on without any extracorporeal perfusion, the VSD was closed by a patch which was attached firmly to the margin of VSD using cross mattress sutures and the right ventricular outflow tract was dilated by infundibular myectomy only, with no use of outflow-patch-plasty. One hundred ten out of 350 patients were investigated by questionnaire and their responses showed that they had few long-term problems. ECG, X-ray, echocardiography and stress test by treadmill were performed in 26 cases. They were divided into two groups by the presence or absence of pulmonary regurgitation (PR). The CTR of the PR (+) group was larger than the PR (-) group (p less than 0.05). There was no statistical difference in cardiac function between the two groups. Few arrhythmias were observed during the exercise test. No residual shunts were found in any of the cases. The occurrence of PR was relatively rare in those who were operated on before their second birthdays. These results suggest that our surgical techniques under simple deep hypothermia are adequate to correct TOF.

Adult↗

[Effectiveness of synthetic calcitonin derivative (elcatonin) on the bone pain and serum calcium concentration in multiple myeloma].

Fifteen patients (8 male and 7 female) with multiple myeloma, who were admitted to our hospital between July 1986 and August 1988 and suffering from pain and hypercalcemia, were treated with synthetic calcitonin derivative (elcatonin: ECT). ECT was administered intravenously at a dose of 10-640 units twice daily. Seven patients were treated with ECT (ECT group), and eight patients received combination treatment with ECT and other form of chemotherapy (combination group). With regard to the pain score (PS), significant analgesic effects in both groups were observed during 1-4 week treatments (p less than 0.05). There were no significant differences in PS between two groups. Serum calcium levels in the combination group at 1 and 4 weeks were significantly lower than the initial value (p less than 0.05). Hypocalcemia was not seen in any of the patients. Urinary excretion of calcium at 1 week in ECT group was higher than the initial value (p less than 0.05). The observed toxicities of ECT were slight nausea and vomiting in only 2 patients. These findings suggest that ECT is an useful agent for the treatment of pain and hypercalcemia accompanied with multiple myeloma.

Aged↗

[Midluteal progesterone/estradiol ratio as an indicator of pregnancy potential].

In an effort to determine the reliability of the midluteal progesterone(P)/estradiol(E2) ratio as an index of the potential for conception, we measured the midluteal P and E2 levels in 76 infertile women who had been treated at our infertility clinic. This parameter in conception cycles was compared with that in non-conception cycles. Eighty cycles of the 76 women were classified into two groups, depending upon whether pregnancy occurred or not. Group 1 and group 2 were composed of 31 conception cycles and 49 drug-induced cycles, respectively. Midluteal concentrations of P and E2 did not show any significant differences between the two groups. The P/E2 ratios were 106.4 +/- 71.3 (mean +/- SD) and 71.5 +/- 44.16, respectively. This difference was statistically significant (p less than 0.05). Discriminant function analysis showed that the smallest probability of misclassification between the two groups decreased from 44% when using P only to 36% when using the combined P and E2. The following equation: Y = 0.0727X1 - 0.00456X2 - 0.130 was obtained (when X1 = P, X2 = E2). These results suggest that the midluteal P/E2 ratio gives clinicians the best indication of luteal function for the achievement of pregnancy.

Adult↗

Vasoactive intestinal peptide immunoreactive neurons in the rat suprachiasmatic nucleus demonstrate diurnal variation.

The suprachiasmatic nucleus (SCN) of mammals is considered to be a circadian oscillator and it also demonstrates circadian rhythmicity of its multiple unit activity. A number of neuropeptides have been found in the SCN. Vasoactive intestinal peptide (VIP)- and vasopressin-containing neurons comprise large populations of these cells and have a distinct distribution within the nucleus. Therefore we attempted to examine whether the VIP neurons show a diurnal alteration of their immunoreactivity by combined immunocytochemistry and color image analysis. Our results demonstrate that VIP-like immunoreactive neurons show a diurnal change in the amount of immunoreactivity. Immunoreactivity was most intense in the sections from rats maintained in the cyclic photoperiod and sacrificed at 02.00 h and weakest in the SCN from animals sacrificed at 14.00 h. We considered that VIP-like immunoreactive neurons showed diurnal variation of VIP synthesis depending strongly on the light from the retina.

Animals↗

Demonstration of GABAergic cell bodies in the suprachiasmatic nucleus: in situ hybridization of glutamic acid decarboxylase (GAD) mRNA and immunocytochemistry of GAD and GABA.

The existence of GABAergic neurons in the rat suprachiasmatic nucleus (SCN) was demonstrated by three specific markers; mRNA coding for glutamic acid decarboxylase (GAD) and visualized by in situ hybridization using a 35S-labelled cDNA probe, and GAD protein and GABA were identified by immunocytochemistry using specific antisera. In situ hybridization demonstrated well labelled GAD mRNA positive cells throughout SCN, and GABA and GAD immunoreactive cells showed similar distributions. These results indicate that GABA is a transmitter of a large portion of the SCN neuronal population.

Animals↗

Purification and protein sequence analysis of rat liver prolactin receptor.

Prolactin receptors were purified from rat liver membranes by single-step immunoaffinity chromatography using a specific monoclonal antibody to the rat liver prolactin receptor. Scatchard analysis of 125I-human growth hormone binding to the purified receptor revealed two classes of specific binding sites with Ka = 18.5 x 10(9) and 1.2 x 10(9) M-1. Considering that both classes of binding sites are responsible for high affinity prolactin binding, the partially purified receptor preparation had a binding activity of 1.69 nmol/mg protein, representing 1000-fold purification over microsomal receptors with a recovery of 52%. From three separate purifications, 6 mg of partially purified prolactin receptor were obtained with a purity of approximately 4 to 6.5%. Thus, the use of monoclonal antibody for affinity chromatography resulted in a large improvement of prolactin receptor purification compared to previous hormone affinity chromatography (300-fold purification, 15% recovery). The purified receptor was run on preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis, and a homogeneous preparation of prolactin receptor was obtained by electroelution from gel slices corresponding to Mr 38,000-43,000. Immunoblot analysis using a radiolabeled monoclonal antibody revealed two separate but closely located bands of Mr 42,000 and 40,000 in microsomal, partially purified, and electroeluted preparations. The homogeneous receptor protein was extensively digested with L-1-tosylamido-2-phenylethyl chloromethyl ketone trypsin, and 10 internal amino acid sequences of the rat liver prolactin receptor were determined by gas-phase sequence analysis. Oligonucleotide probes were prepared against two of these internal sequences, and a prolactin receptor cDNA was isolated from a rat liver library using one of these probes (Boutin, J. M., Jolicoeur, C., Okamura, H., Gagnon, J., Edery, M., Shirota, M., Banville, D., Dusanter-Fourt, I., Djiane, J., and Kelly, P. A. (1988) Cell 53, 69-77). The amino acid sequence deduced from the cDNA reveals three potential sites of N-linked glycosylation, two of which were confirmed during protein sequencing. The prolactin receptor was characterized by affinity labeling with 125I-human growth hormone. Cross-linking of microsomes revealed a single band for the hormone-receptor complex with Mr 62,000. On the other hand, cross-linking of Triton X-100-solubilized or partially purified receptor with labeled hormone resulted in the appearance of two bands with Mr 62,000 and 102,000, suggesting the existence of a subunit structure of the prolactin receptor, or alternatively, the existence of two types of prolactin receptor.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Differential colocalization of neuropeptide Y- and methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactivity in catecholaminergic neurons in the rat brain stem.

The present study, using a combination of catecholamine (CA) histofluorescence and peptide immunocytochemistry in the same tissue sections, investigated the coexistence of neuropeptide Y (NPY) and methionine-enkephalin-Arg6-Gly7-Leu8 (MEAGL)-like immunoreactivity (LI) in catecholaminergic neurons of colchicine-treated rat brain stems. Of the total number of catecholaminergic neurons in the A1/C1, A2/C2, A3, A4, and A6 regions approximately 83, 28, 98, 76, and 36%, respectively, contained both NPY-LI and CA. Of the total number of catecholaminergic neurons in A1/C1, A2/C2, A3, and A5 regions, approximately 47, 4, 8, and 17%, respectively, contained both MEAGL-LI and CA. Moreover, about 24% of the catecholaminergic neurons in the A1/C1 region contained both NPY- and MEAGL-LI. Neither the noradrenergic neurons (A7) in the pons nor any of the dopaminergic neurons in the midbrain (A8, A9, A10) contained NPY- or MEAGL-LI. Neurons containing both NPY- and MEAGL-like immunoreactive peptides without CA were not found in the rat brain stem. These findings indicate that catecholaminergic neurons in the brain stem of the rat can be subdivided into distinct subgroups on the basis of the coexistence of specific peptides.

Animals↗

Vasoactive intestinal peptide (VIP)-like immunoreactive neurons located in the rat suprachiasmatic nucleus receive a direct retinal projection.

The existence of a direct projection from retinal ganglion cells to vasoactive intestinal peptide (VIP)-like immunoreactive neuronal elements in the rat suprachiasmatic nucleus (SCN) was revealed by combining analysis of degenerating axons following enucleation and electron microscopic immunocytochemistry. Degenerating axons appeared to make synaptic contact with VIP-like immunoreactive dendrite and neuronal perikarya in the ventral part of the SCN. The possibility of neuronal input from retinal ganglion cells to axons of VIP-like immunoreactive neurons was also suspected since axo-axonic synapses were detected between degenerating axons and axons with VIP-like immunoreactivity. Thus, VIP-like immunoreactive neurons in the SCN receive several neuronal inputs, including those from the retina, and may play a significant role in circadian entrainment.

Animals↗

A new group of tyrosine hydroxylase-immunoreactive neurons in the cat thalamus.

A new cell group composed of a large number of neurons immunoreactive to tyrosine hydroxylase (TH) was demonstrated in the paraventricular nucleus and midline nuclei of the cat thalamus, using four different anti-TH sera after colchicine treatment. However, in these regions, we did not detect any cell bodies containing other catecholamine synthesizing enzymes nor dopamine.

Animals↗

Comparative distribution of three opioid systems in the lower brainstem of the monkey (Macaca fuscata).

The regional distribution of the three opioid peptide neuronal systems--proopiomelanocortin (POMC), proenkephalin A, and proenkephalin B--was investigated in the lower brainstem of Japanese monkeys (Macaca fuscata) by immunocytochemical techniques. Antiserum to beta-endorphin/beta-lipotropin, [Met]-enkephalin-Arg6-Gly7-Leu8, and human leumorphin were used to identify the POMC and the proenkephalin A and B systems, respectively. POMC-related immunoreactive material was not found in the neuronal perikarya in the lower brainstem; reactive fibers and apparent terminals were distributed in the substantia nigra, lemniscus lateralis, midbrain central gray, the nucleus raphes, nucleus parabrachialis lateralis, ventral area of the spinal trigeminal nerve, nucleus tractus solitarii, and in the reticular formation throughout the lower brainstem. Proenkephalin A-related immunoreactive neuronal perikarya were detected in the central gray, reticular formation, nucleus raphes, trapezoid body, nucleus parabrachialis lateralis and medialis, nucleus spinalis nervi trigemini, nucleus dorsalis nervi vagi, and in the nucleus tractus solitarii. Densely packed immunoreactive fibers were widely distributed in the substantia nigra, nucleus interpeduncularis, nucleus raphes, superior colliculus, periaqueductal central gray, nucleus parabrachialis lateralis and medialis, locus coeruleus, trapezoid body, nuclei cochleares, nucleus spinalis nervi trigemini, tractus spinalis nervi trigemini, nucleus tractus solitarii, nucleus dorsalis nervi vagi, nucleus gracilis, nucleus cuneatus, nucleus cuneatus accessorius, and in the reticular formation throughout the lower brainstem. Neuronal perikarya containing immunoreactive material related to proenkephalin B were found in the periaqueductal central gray, nucleus parabrachialis lateralis and medialis, nucleus tractus solitarii, and nucleus spinalis nervi trigemini. In addition, immunoreactive fibers were detected in the ventral tegmental area, substantia nigra, nucleus parabrachialis lateralis and medialis, nucleus vestibularis lateralis and medialis, and in some areas of the reticular formation. These anatomical findings demonstrate that these three opioid peptide neuronal systems are widely but uniquely distributed in the lower brainstem of the monkey.

Animals↗

Coexistence of catecholamine and methionine enkephalin-Arg6-Gly7-Leu8 in neurons of the rat ventrolateral medulla oblongata. Application of combined peptide immunocytochemistry and histofluorescence method in the same vibratome section.

An overlapping distribution of catecholamine-containing cells and proenkephaline-A derived peptide-containing neurons have been identified in the rat medulla oblongata. However, it is not evident whether the coexistence of these bioactive substances occurs in the same neurons or not. Therefore, we examined the coexistence of catecholamine and methionine-enkephalin-Arg6-Gly7-Leu8 (MEAGL), a proenkephaline-A derived peptide, using a combination of histofluorescence and peroxidase-anti-peroxidase (PAP) immunohistochemical (modified formaldehyde-glutalaldehyde (Faglu)) methods on the same tissue sections. We found one third of A1/C1 catecholamine fluorescent cells show MEAGL-like immunoreactivity.

Animals↗

Caudato-nigral transmission in the substantia nigra pars reticulata neurons changes with recovery from circling movements induced by microinfusion of ibotenic acid. I. Behavioral study.

Following creation of a unilateral circumscribed lesion in a portion of a cat substantia nigra pars reticulata by microinfusion of ibotenic acid, circling movements toward the contralateral side of the lesion appeared within 2 days and disappeared a few days later. After recovery, the circling movements reappeared when the cats were decerebrated at the premammillary and precollicular level, suggesting that brain centers rostral to the decerebration participate in compensating the circling movements.

Animals↗

Immunocytochemical localization, binding, and effects of atrial natriuretic peptide in rat adipocytes.

The metabolic effects of atrial natriuretic peptide (ANP) have not been widely investigated. Since adipocyte cells represent a model system extensively used to examine the metabolic actions of many peptide hormones, we sought to establish whether ANP could bind to adipocyte membranes, alter cyclic nucleotide metabolism, and affect spontaneous or hormone-stimulated lipolysis. Using in vitro autoradiographic techniques, radiolabelled ANP was found to bind specifically to mammary gland fat cells. Additionally, endogenous ANP-like immunoreactivity could be localized in the plasma membrane compartment and cytoplasmic matrix of fat cells, but not in fat vacuoles. [125I]ANP bound to single high affinity sites (Kd = 0.72 nM) in fat cell membranes. The binding was rapid (equilibrium within 1 min at 25 degrees C) and specific. The atrial peptide was capable of stimulating a time- and concentration-dependent increase in cGMP accumulation in isolated adipocytes, but had no effect on spontaneous or stimulated [-)-isoproterenol, ACTH, forskolin) cAMP formation. ANP did not alter the increase in glycerol production stimulated by l-epinephrine in isolated fat cells. While i.v. infusion of ANP stimulated a marked increase in circulating levels of cGMP, the atrial peptide did not alter plasma triglyceride levels. These data demonstrate the presence of specific ANP binding sites on adipocyte membranes and internalization of ANP-associated immunoreactivity. These receptors are biochemically functional given the ability of ANP to augment cGMP formation. The peptide, however, does not exert an action on adipocyte lipolysis. Adipocytes, therefore, represent an ANP target tissue in which the physiological action of the peptide is yet to be defined.

Adipose Tissue↗

Effects of pH and ionic strength on the binding of egg white riboflavin binding protein with flavins.

The effects of pH and ionic strength on the equilibrium constants and rate constants (binding and dissociation rate constants) between riboflavin binding protein (RBP) and flavins (riboflavin, 3-carboxymethylriboflavin [CMRF], and FMN) were studied by fluorometry. The equilibrium constant and the binding rate constant between RBP and riboflavin were pH-independent between pH 6 and 9, and both constants were also independent of the ionic strength, while the constants between RBP and CMRF or FMN were dependent on both pH and ionic strength. The dissociation rate constants between RBP and the flavins used here were not so dependent on pH and ionic strength in the pH region 6 to 9, and the patterns of pH profiles as a whole were similar to each other, although the constants for FMN were about 30-60 times larger than those for CMRF or riboflavin. RBP had lower affinity for FMN than for riboflavin in the neutral pH region, which is based on the small binding rate constant and the large dissociation rate constant for FMN. The former is due to an electrostatic repulsion force between negative net charges of RBP and the phosphate group of FMN, and the latter is due to steric interference by the phosphate group of FMN.

Animals↗

Purification, cloning, and expression of the prolactin receptor.

The rat liver prolactin receptor has been purified to homogeneity, and partial amino acid sequences have been obtained. The structure of the receptor has been deduced from a single complementary DNA clone. The mature protein of 291 amino acids has a relatively long extracellular region, a single transmembrane segment, and a short (57 amino acids) cytoplasmic domain. With the rat cDNA used as a probe, the prolactin receptor in rabbit mammary gland and human hepatoma cells has also been isolated. These tissues contain a second, longer form of the receptor (592 and 598 amino acids, respectively). Both the short and long forms of the prolactin receptor show regions of strong sequence identity with the human and rabbit growth hormone receptors, suggesting that the prolactin and growth hormone receptors originate from a common ancestor.

Amino Acid Sequence↗

Increase in ovarian blood volume during ovulation in the gonadotropin-primed immature rat.

This study quantifies ovarian blood volume in Wistar rats by measuring the optical density (414 nm) of hemoglobin in ovarian extracts and comparing this measurement to the optical density of known amounts of whole blood. Immature rats were primed with pregnant mare's serum gonadotropin (PMSG), 10 IU s.c., at 23 days of age. On Day 25, the ovulatory process was initiated by human chorionic gonadotropin (hCG), 10 IU s.c., and ova began to appear in the oviducts 10 h later. At 2-h intervals, the ovaries were extirpated and homogenized in 1.0 ml of 0.05 M tris (hydroxymethyl)aminomethane buffer (pH 7.4) for 30 s. Homogenates were centrifuged for 20 min and the supernatant fluids were analyzed with a Gilford RESPONSE UV/VIS spectrophotometer. The hemoglobin in these ovarian extracts had the same peak absorbance of 414 nm characteristic of oxyhemoglobin in whole blood taken by cardiac puncture of the rats. There was a linear relationship between the absorbance and the volume of whole blood in the samples. The volume of blood per ovary from groups of 8 rats was 0.60 +/- 0.07 microL at 0 h after hCG. The volume increased to 1.37 +/- 0.26 microL at 4 h after hCG and reached a peak of 4.55 +/- 0.72 microL at 10 h. Indomethacin treatment (0.3-10.0 mg/rat, s.c.) partially inhibited this 7-fold increase in ovarian blood volume. In conclusion, the increase in ovarian blood volume during ovulation may reflect the vasodilation and hyperemia that are characteristic of inflamed tissues.

Animals↗

Decrease in ovarian platelet-activating factor during ovulation in the gonadotropin-primed immature rat.

Platelet-activating factor (PAF) is a biologically active phospholipid that is released locally during acute inflammatory reactions and tissue injury. Since there is evidence that the biochemical events of mammalian ovulation resemble an inflammatory reaction, the objective of this study was to determine whether ovarian levels of PAF change during ovulation. At 2-h intervals during the ovulatory process in gonadotropin-primed 25-day-old Wistar rats, the ovaries were extirpated, homogenized, and extracted for lipids. The extracts were subjected to thin-layer chromatography (TLC), and the portion of the silica gel that comigrated with PAF was re-extracted and assayed for PAF activity. The PAF was measured (in fmole equivalents of synthetic PAF) by a bioassay based on the capacity of aliquots of the extracts to release [3H]-serotonin from platelets isolated from whole blood of rabbits and prelabeled with [3H]-serotonin. The ovarian level of PAF decreased (p less than 0.01) by 36% from 6.67 +/- 0.77 to 4.27 +/- 0.45 fmoles/mg ovary by 2 h after treatment with human chorionic gonadotropin (hCG), and it declined another 14% by 4 h after hCG. The ovarian PAF remained at this reduced level for up to 24 h after hCG. The administration of indomethacin (5 mg/rat, s.c.) or epostane (5 mg/rat, s.c.) at 1 h after hCG prevented ovulation, but neither drug affected the decline in ovarian PAF. Preliminary tests showed that the lipid extracts from the ovaries also contained PAF inhibitor(s) that comigrated with PAF on the TLC plates. Similar to PAF, the lipid-soluble inhibitor(s) decreased (p less than 0.05) in the ovaries within 4 h after hCG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Androstenols↗