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Biomedical subjects

A Sato

Publications and source records attributed to A Sato.

At least 19 recordsLinked to original sources

Responses of regional cerebral blood flow to intravenous administration of thyrotropin releasing hormone in aged rats.

The effects of i.v. administration of thyrotropin releasing hormone (TRH) on regional cerebral blood flow (rCBF) were examined in both healthy adult (3-5 months old) and healthy aged (24-25 months old) male Wistar rats under halothane anesthesia. The rCBFs in 9 different brain regions-cerebral cortex, caudate putamen, hippocampus, thalamus + hypothalamus, superior colliculus, inferior colliculus, cerebellum, pons, and medulla-were measured by [14C]iodoantipyrine method. In the adult rats, i.v. administration of TRH (300 micrograms/kg) produced significant increases in rCBFs in cerebral cortex, caudate putamen, hippocampus, thalamus + hypothalamus and superior colliculus. In the aged rats, the rCBFs in all brain regions measured did not change significantly by TRH administration. From these results, it is suggested that the system involved in TRH-induced vasodilatation of cerebral blood vessels was impaired with aging.

Aging

Stimulation of the nucleus basalis of Meynert increases cortical cerebral blood flow without influencing diameter of the pial artery in rats.

The effects of focal electrical stimulation of the nucleus basalis of Meynert (NBM) and hypercapnia on diameter of the pial artery and regional cerebral blood flow (CBF) in the parietal cortex were examined in urethane-anesthetized rats. The diameter of pial artery was measured using the video microscope and cortical CBF was measured using the laser Doppler flowmetry. Hypercapnia at 10% of end-tidal CO2 concentration caused significant increases in diameter of the pial artery and cortical CBF. Electrical stimulation of the NBM (with parameters of 200 microA, 50 Hz, 0.5 ms, for 1 min) produced a significant increase in cortical CBF, but did not influence the diameter of pial artery. The results suggest that cholinergic fibers originating in the NBM do not contribute to neural vasodilatation of the pial artery, although they induce neural vasodilation of parenchymal blood vessels which results in an increase in CBF in the parietal cortex.

Animals

Construction of a tRNA-embedded-ribozyme trimming plasmid.

We have combined Cotten and Birnstiel's tRNA-embedded ribozymes and our 5'- and 3'-trimming system. Although the activity of the tRNA-embedded ribozyme was ca. 30% lower than those of naked ribozymes, since the stability of the former in bovine serum was higher than those of naked ribozymes, the tRNA-embedded ribozymes appear useful especially when the 5'- and 3'-trimming units are concatenated in tandem.

Base Sequence

A farnesyl arabinoside as an enhancer of glucose transport in rat adipocytes from a soft coral, Sinularia sp.

Separation of a lipophilic extract of a soft coral, Sinularia sp., assayed by enhancement of glucose transport in rat adipocytes, gave farnesyl 4-O-beta-D-arabinopyranosyl-beta-D-arabinopyranoside-2,2',3- triacetate (1a) whose structure was determined by spectroscopy. Enhancers of glucose transport may be useful for the prevention and treatment of diabetic disorders.

Adipose Tissue

Design and synthesis of a CD4 beta-turn mimetic that inhibits human immunodeficiency virus envelope glycoprotein gp120 binding and infection of human lymphocytes.

Poor bioavailability, rapid degradation, antigenicity, and high cost often limit the use of proteinaceous pharmaceuticals. One goal of structural biochemistry is the reduction of complex molecules to small functional units that are amenable to high-resolution structural analysis and rapid modification. The dissection of complex proteins into small synthetic conformationally restricted components is an important step in the design of low molecular weight nonpeptides that mimic the activity of the native protein. We have developed a reverse-turn mimetic system to explore peptide and protein structure-function relationships. We now report the design and synthesis of a small molecule (M(r) 810, as its trifluoroacetate salt), water soluble, proteolytically stable mimetic of residues Gln40-Thr45 of the complementarity-determining 2-like region of CD4. This mimetic has a low micromolar Kd for human T-lymphotropic virus type IIIB gp120 and reduces syncytium formation.

Binding, Competitive

Nitric oxide (NO) is involved in increased cerebral cortical blood flow following stimulation of the nucleus basalis of Meynert in anesthetized rats.

The effects of i.v. administration of a nitric oxide (NO) synthase inhibitor, L-NG-nitroarginine (L-NOArg), on the increase in cerebral cortical blood flow (cortical BF), following either electrical stimulation of the nucleus basalis of Meynert (NBM), whose cholinergic fibers project to the cortex, or hypercapnia with 10% CO2 inhalation, were studied in anesthetized rats. Cortical BF was measured using laser Doppler flowmetry. The threshold intensity of electrical stimulation of the NBM (0.5 ms, 50 Hz for 10 s) that induced an increase in regional cortical BF was defined as 1T. The cortical BF was increased on a stimulus intensity dependent manner at 1T-5T intensities tested. L-NOArg was administered cumulatively i.v. starting from 0.3 mg/kg, then 3 mg/kg, and 30 mg/kg. Time interval between each cumulative administration of L-NOArg was approximately 40 min. Three and 30 mg/kg of L-NOArg significant reduced the NBM stimulation-induced increase of cortical BF at intensities of 2T and 3T. The response at an intensity of 5T was reduced only by 30 mg/kg of L-NOArg to about half the control response. The reduced responses at 2T, 3T, and 5T were reversed following the i.v. administration of a physiological precursor of NO, L-arg (300 mg/kg). Inhalation of 10% CO2 for 15 s induced an increase in cortical BF which was not influenced by L-NOArg and L-Arg. These results suggest that NO is a necessary factor in the vasodilation of the cortical BF that is brought about by cholinergic fibers originating in the NBM.

Anesthesia

The effects of morphine administered into the vertebral artery on the somatosympathetic A- and C-reflexes in anesthetized cats.

The effects of morphine (0.2-200 micrograms/kg) administered into the brain stem via a vertebral artery on the somatosympathetic A- and C-reflexes evoked in a cardiac sympathetic nerve by electrical stimulation of somatic myelinated (A) and unmyelinated (C) afferent nerve fibers in anesthetized cats were examined. The intravertebral arterial administration of 20-200 micrograms/kg morphine depressed dose-dependently both the somatosympathetic A- and C-reflexes. Following the administration of 200 micrograms/kg morphine, the amplitudes of the sympathetic A- and C-reflexes were depressed to 71 +/- 8% and 80 +/- 8% of the control magnitude, respectively. Injection of 2-200 micrograms/kg morphine caused hypotension in a dose-dependent manner. It is concluded that morphine administered in the brain stem via the vertebral artery produced a nonselective and slight depression of both the somatosympathetic A- and C-reflexes by acting at the level of brain stem.

Anesthesia

Involvement of lysosomes in substrate stabilization of tryptophan-2,3-dioxygenase in rat liver.

Administration of tryptophan or hydrocortisone to rats caused a several-fold increase in tryptophan-2,3-dioxygenase activity in the liver. Highly purified lysosomes were isolated from livers of tryptophan- or hydrocortisone-treated animals as well as the control rats. Immunoblotting of lysosomal proteins with anti-tryptophan-2,3-dioxygenase showed 48 kDa band, corresponding to the subunit molecular weight of the enzyme. The relative amount of the immuno-reactive substance in the lysosomes from hydrocortisone-treated rats was 3 times higher than the control while the value in the lysosomes from tryptophan-treated rats was essentially the same as in the control. These results indicate that administration of tryptophan renders cytosolic tryptophan-2,3-dioxygenase less vulnerable to lysosomal uptake and causes an accumulation of the enzyme in the cytosol.

Animals

Enhancement of ethanol-induced lipid peroxidation in rat liver by lowered carbohydrate intake.

In order to investigate the effect of carbohydrate intake on ethanol-induced lipid peroxidation and cytotoxicity, rats were maintained on four different test diets, a medium-carbohydrate (carbohydrate intake, 8.4 g/day/rat on average), a low-carbohydrate (carbohydrate intake, 2.8 g/day/rat on average), an ethanol-containing medium-carbohydrate (carbohydrate and an ethanol intake, 8.4 and 2.9 g/day/rat on average, respectively), and an ethanol-containing low-carbohydrate diet (2.8 and 2.9 g/day/rat on average, respectively). Ethanol and the low-carbohydrate diet each increased the liver malondialdehyde content, but the combined effect of both (ethanol-containing low-carbohydrate diet) was much more prominent than either alone. The degree of increase in malondialdehyde content almost paralleled the activity of the microsomal ethanol oxidizing system. Both the low-carbohydrate and the ethanol-containing low-carbohydrate diets decreased the liver glutathione content, but ethanol combined with the medium-carbohydrate diet had no effect on the content. Ethanol treatment increased the liver triglyceride content only when combined with the low-carbohydrate diet. The rate of NADPH-dependent microsomal malondialdehyde formation was much higher in microsomes from rats maintained on the ethanol-containing low-carbohydrate diet than in those from rats on the ethanol-containing medium-carbohydrate diet, indicating that lowered carbohydrate intake augments ethanol-induced malondialdehyde accumulation in the liver by enhancing the rate of lipid peroxidation. In addition, when incubated with red blood cells in the presence of NADPH, microsomes from rats fed the ethanol-containing low-carbohydrate diet caused marked hemolysis, which was prevented by the addition of 5 mM glutathione to the incubation system. Furthermore, addition of 50 mM ethanol to the reaction system greatly accentuated the hemolysis. These results suggest that lowered carbohydrate intake at the time of ethanol consumption potentiates ethanol cytotoxicity by enhancing ethanol-induced lipid peroxidation.

Animals

Non-essential repeats in the promoter region of a Brassica rapa acyl carrier protein gene expressed in developing embryos.

A genomic clone of an acyl carrier protein gene (Bcg4-4) which is highly expressed in developing embryos of Brassica rapa was isolated and sequenced. The promoter and transcription terminator regions of Bcg4-4 were used to express a beta-glucuronidase reporter gene in transgenic rapeseed. Deletion of repeated domains in the promoter region did not lower beta-glucuronidase expression in seeds.

Acyl Carrier Protein

Occurrence of unusual heterogeneous lipid-containing granule-storing cells in the main excretory duct epithelium of the male mouse submandibular gland.

The fine structure of the main excretory duct epithelium of the male mouse submandibular glands was investigated by scanning and transmission electron microscopy. Three principal cell-types were observed: type I and II, and basal cells. This epithelium was characterized by the presence of intercellular canaliculi. Type-I cells were the most numerous. They had an abundance of mitochondria, well-developed Golgi apparatus, a few electron-lucent lipid-containing granules and poorly developed basal infoldings. These cells were also characterized by many glycogen granules throughout the cytoplasm and abundant smooth endoplasmic reticulum in the apical cytoplasm. Type-II cells were the second most numerous. Their most characteristic feature was the presence of abundant heterogeneous lipid-containing granules having acid phosphatase activity at the periphery. They were concentrated in the infra- and supra-nuclear cytoplasm. The granules may be derived from mitochondrial transformation and seem to be a special kind of secondary autolysosome. Type-II cells also contained abundant mitochondria throughout the cytoplasm, much smooth endoplasmic reticulum in the apical cytoplasm, a well developed Golgi apparatus adjacent to the heterogeneous lipid-containing granules and no basal infoldings. Basal cells were situated adjacent to the basal lamina. They had a large nucleus and the cytoplasm was filled with glycogen granules.

Acid Phosphatase

Rapid, sensitive, specific, and quantitative detection of human T-cell leukemia virus type 1 sequence in peripheral blood mononuclear cells by an improved polymerase chain reaction method with nested primers.

Improving on the nested double polymerase chain reaction (PCR) described previously, we have developed a new two-step PCR (TS-PCR) method for detecting more specifically the human T-cell leukemia virus type 1 (HTLV-1) proviral sequences in peripheral blood mononuclear cells (PBMC). In our TS-PCR method, the point of modification is to use optimal concentrations of primers in the first amplification step in the range of 0.01-0.025 microM. This increases sensitivity and specificity enough to detect from 1 to 10(5) copies of template DNA without radioisotopes. This method is rapid because of completion in 1 day and is also applicable for quantitative detection of clinical specimens. The data show that the quantitative detection of HTLV-1 proviral sequences by this method correlates with the anti-HTLV-1 antibody titers from serologic analysis of seropositive healthy carriers. Moreover, the TS-PCR method using each specific primer was also attempted for successful detection of other viral genomes; therefore, the principle of this method is widely suitable for routine detection of genomes in the basic and clinical microbiological fields.

Base Sequence

Multiple endocrine neoplasia type 2 with malignant pheochromocytoma--long term follow-up of a case by 131I-meta-iodobenzylguanidine scintigraphy.

The case of a 33-year-old Japanese man, who has Multiple Endocrine Neoplasia Type 2 (MEN IIa) (Sipple's syndrome) with malignant pheochromocytoma, is reported. He had survived for twelve years since the initial diagnosis of malignant pheochromocytoma. Within this period, he had undergone 131I-meta-iodobenzylguanidine scintigraphy twice, in 1983 and 1990. This is the first case in Japan of a longterm surviving patient with malignant pheochromocytoma followed up by 131I-MIBG scintigraphy. Although he had no exacerbation of clinical symptoms or urinary catecholamine levels, second scintigraphy clearly showed an increase in the tumor size, new metastasis of the malignant pheochromocytoma and exacerbation of the medullary thyroid carcinoma. Compared with any other roentgenological device and hormonal data, 131I-MIBG scintigraphy was seen to be a good tool for evaluating the localization and the progression of tumors. 131I-MIBG scintigraphy is a useful procedure not only for initial diagnosis but also for judging progression in a case of advanced malignant pheochromocytoma.

3-Iodobenzylguanidine

Local cerebral blood flow responses in rats to hypercapnia and hypoxia in the rostral ventrolateral medulla and in the cortex.

The effects of hypercapnia and hypoxia on two local cerebral blood flows in the parietal cortex (PC-BF) and rostral ventrolateral medulla (RVLM-BF) were examined using laser Doppler flowmetry in anesthetized rats. Hypercapnia for 45 s duration at the end-tidal CO2 between 5% and 10%, induced by increasing inspiratory CO2, increased both cerebral blood flows and systemic blood pressure in a degree-dependent manner. The response of RVLM-BF was significantly stronger than that of PC-BF. Both cerebral blood flow responses to hypercapnia were not influenced by cutting peripheral chemoreceptor afferent nerves. Hypoxia for 45 s duration at the end-tidal O2 between 12% and 6%, induced by decreasing inspiratory O2, produced an increase of similar magnitude in both RVLM and PC local blood flows in a degree-dependent manner and a decrease in systemic blood pressure. The responses of both PC-BF and RVLM-BF to hypoxia were significantly diminished after cutting peripheral chemoreceptor afferent nerves. It is concluded that: (1) the RVLM-BF is much more sensitive to hypercapnia than the PC-BF; and (2) activation of peripheral arterial chemoreceptors possibly contributes to hypoxia-induced increase in the RVLM-BF and PC-BF.

Animals

Mechanism of the reflex inhibition of micturition contractions of the urinary bladder elicited by acupuncture-like stimulation in anesthetized rats.

The effects of acupuncture-like stimulation of various segmental areas on the rhythmic micturition contractions (RMCs) of the urinary bladder were examined in anesthetized rats. The urinary bladder was cannulated via the urethra and expanded by infusing saline until the urinary bladder produced micturition contractions rhythmically as a consequence of the rhythmic burst discharges of the vesical pelvic efferent nerves. An acupuncture needle, having a diameter of either 160 or 340 microns, was inserted to a depth of about 4-5 mm into the skin and underlying muscles at various segmental areas, rostrally from the face then caudally to the hindlimb. Once being inserted, the needle was twisted left and right with the fingers about once every second for 1 min. (1) Acupuncture-like stimulation applied to the perineal area inhibited both the RMCs and the rhythmic burst discharges of vesical pelvic efferent nerves without any significant changes in the hypogastric efferent nerve activity. By contrast, stimulation applied to the face, neck, forelimb, chest, abdomen, back, and hindlimb areas was ineffective. (2) After surgically separating the perineal skin from the underlying muscles with the main cutaneous nerve branches intact, stimulation of either the perineal skin or the perineal muscles inhibited the RMCs. Stimulation of the perineal muscles produced a stronger inhibition of the RMCs than that of the perineal skin. (3) Stimulation of the perineal area increased afferent nerve activity, either recorded from the pudendal nerve branches innervating the perineal skin or underlying muscles, or recorded from the pelvic nerve branches innervating the perineal muscles. (4) The stimulation-induced inhibition of the RMCs was abolished after surgically severing both pudendal and pelvic nerve branches that innervated the perineal skin and underlying muscles. (5) The present findings indicate that the inhibition of the RMCs following acupuncture-like stimulation of the perineal area is a reflex response characterized by segmental organization. The afferent arcs of the reflex are both pelvic and pudendal nerve branches innervating the perineal skin and underlying muscles, while the efferent arcs are pelvic nerve branches innervating the urinary bladder.

Acupuncture Therapy

Depending on the mode of application morphine enhances or depresses somatocardiac sympathetic A- and C-reflexes in anesthetized rats.

The effects of morphine on the reflex discharges in sympathetic efferents recorded from branches of the inferior cardiac nerve (ICN) were studied in rats anesthetized with alpha-chloralose and urethane. Somatocardiac sympathetic A- and C-reflexes were elicited by single shock electrical stimulation of myelinated (A) and unmyelinated (C) afferent fibers of the tibial nerve, respectively. Application of morphine either into the femoral vein or into the subarachnoid space of the cisterna magna enhanced both the A- and C-reflexes in a dose-dependent manner, while application of morphine into the intrathecal space of the lumbar spinal cord selectively inhibited C-reflexes. All effects of morphine were antagonized by naloxone. Application of morphine via the internal carotid artery to central nervous structures above the brainstem had no effect on the somatocardiac sympathetic reflexes. It is concluded that in the anesthetized rat morphine in a dose-dependent and naloxone-reversible manner (1) depresses spinal transmission of C-afferent activity, whereas (2) at the brainstem it enhances the transmission of somatocardiac sympathetic A- and C-reflexes.

Adrenergic Fibers

Regulation of regional cerebral blood flow by cholinergic fibers originating in the basal forebrain.

We review mainly recent studies on vasodilative regulation of cortex and hippocampus by central cholinergic nerves originating in the basal forebrain. We also briefly review the influence of other central noradrenergic fibers originating in the locus ceruleus, serotonergic fibers originating in the dorsal raphe nucleus, dopaminergic fibers originating in the substantia nigra, and peripheral sympathetic and parasympathetic nerve fibers upon regulation of regional cerebral blood flow. Local metabolites have long been considered to play an important physiological role in regulating regional cerebral blood flow. However, the evidence reviewed here emphasizes that the regulation of regional cerebral blood flow by these central cholinergic nerves is independent of regional metabolism. We propose through this review that although studies investigating neural regulation of cortical and hippocampal blood flow by cholinergic fibers originating in the basal forebrain have added much to the understanding of regulation of regional cerebral blood flow further studies are needed to determine the physiological relevance of regional cerebral blood flow in relation to higher nervous functions such as memory, learning, and personality, and changes in these cognitive functions with aging and pathology such as Alzheimer's disease.

Animals