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Biomedical subjects

K Muramoto

Publications and source records attributed to K Muramoto.

At least 109 records · Page 6Linked to original sources

Isolation and characterization of sulfated glycoprotein from human pancreatic juice.

Sulfated glycoprotein was isolated by precipitation from dialyzed human pancreatic juice and purified by ion-exchange chromatography followed by repeated gel chromatography. The sulfated glycoprotein was obtained as a sulfated glycoprotein-lipid complex by Sepharose CL-2B chromatography. Lipids aggregating with the sulfated glycoprotein were glycolipids such as ceramide trihexoside, and simple lipids such as cholesterol and cholesterol ester. This glycoprotein was resistant to digestion with mucopolysaccharidases or alpha-amylase, and consisted of 60% (w/w) protein and 40% sugars. The polypeptide core was characterized by a high content of serine, threonine, aspartic acid and glycine, but lacked cysteine. Its sugar components were N-acetylglucosamine, N-acetylgalactosamine, galactose, fucose and sialic acid. Absorption at 1240 cm-1 and 820 cm-1 by infrared spectroscopy indicated the presence of a sulfate ester group. All the carbohydrate chains of this sulfated glycoprotein, which are polydisperse and heterogeneous, were O-glycosidically linked through N-acetylgalactosamine to a protein core.

Amino Acids↗

Production of an eosinophil chemotactic lymphokine by a monocyte-derived factor from patients with hypereosinophilia.

Peripheral OKT4-positive T lymphocytes from patients with hypereosinophilia spontaneously and selectively produced an eosinophil chemotactic factor (ECF) with chemokinetic activity. The molecular weight of the ECF was about 45,000 to 70,000. A possible mechanism of its spontaneous production by T lymphocytes was analyzed. Culture supernatants of blood monocytes from the patients showed little or no ECF activity, but they had a potency to induce the ECF production from T lymphocytes from normal donors when the cells were stimulated by the supernatants, which suggests that a monocyte-derived soluble factor (MDF) stimulated T lymphocytes to produce an ECF resembling this spontaneously produced ECF from the patients. MDF seemed to be a synthesized protein by the cells. Gel filtration indicated that molecular weight of MDF ranged between 70,000 and 100,000. MDF activity was stable at 56 degrees C for 30 min but more, supernatants of stimulated monocytes by lipopolysaccharide or silica particles failed to show ECF-producing activity, whereas they showed evident lymphocyte-activation activity. Neither recombinant IL-1 nor IL-2 had ECF and ECF-producing activity. From the present experiments, it was suggested that MDF was at least partly involved in the induction of ECF production by OKT4-positive T lymphocytes in patients with hypereosinophilia.

Adult↗

Purification and properties of agglutinins from conger eel, Conger Myriaster (Brevoort), skin mucus.

The skin mucus of the conger eel, Conger myriaster, contains galactose-specific agglutinins. Hemagglutinating activity is independent of divalent cations and is destroyed by heating at 50 degrees C for 15 min. The mucus agglutinins, named congerins, are a mixture of proteins with different electrical charges. Three of these molecules were isolated by affinity chromatography on acid-treated Sepharose 4B and by ion-exchange chromatography. They are simple proteins with the same molecular weight of 30,000 and consisting of two subunits (each 13,000 daltons). The agglutinins inhibited the normal embryonic development of the starfish Asterina pectinifera, and lysed the fertilized eggs at a concentration of 25 micrograms protein/ml. They also agglutinated but did not inhibit the growth of a marine bacterium, Vibrio anguillarum.

Agglutinins↗

Neuronal activity in the ventral tegmental area (VTA) during motivated bar press feeding in the monkey.

Neuronal activity of 58 dopaminergic (DA) and 200 non-dopaminergic (non-DA) neurons in the ventral tegmental area (VTA) of female monkeys was recorded, and correlation to bar press feeding, sensory stimulation and change in motivation was investigated. DA neurons, judged by duration of action potentials (more than 2.5 ms) and responsiveness to apomorphine, had lower firing rates (0-8 impulses/s); non-DA neurons had intermediate firing rates (10-30 impulses/s). Two-thirds of the DA and non-DA neurons responded in bar press feeding; the former with mostly tonic and the latter with phasic responses. Fifteen neurons (5%) responded phasically to arm extension toward the bar, 124 (excitation 88, inhibition 36, 45%) during bar press (BP), and 91 (excitation 32, inhibition 59, 33%) during ingestion reward (RW). Most BP responses (84/124, 68%) continued tonically throughout the BP period with no correlation to each BP movement. In 14 neurons (14/124, 11%), firing showed a specific variation: transient early BP responses shifted to tonic steady ones in palatable food trials, and the shifts correlated well with BP speed. In 20 other neurons, firing increased during BP hip lifting, and at specific vocalization to ask for food; it decreased during food ingestion, drinking and inguino-crural stimulation. Apomorphine administration decreased firing for the first 5-15 min, then increased it with frequent lip smacking, nausea, involuntary movement and vocalization. Thus VTA neurons showed mostly steady tonic responses but some specific phasic responses. They responded not only to motor events but also in close relation to changes of motivational aspects. Neuronal responses were excitation during procurement of reward and inhibition during or after perception of reward. This modulation in firing, might be important in the initiation and execution of movement and/or motivated behavior.

Action Potentials↗

Analysis of reducing sugars as their chromophoric hydrazones by high-performance liquid chromatography.

A chromophoric hydrazide, 4'-N,N-dimethylamino-4-azobenzene sulfonyl hydrazide (DABS-hydrazide), was prepared from 4'-N,N-dimethylamino-4-azobenzene sulfonyl chloride by reaction with hydrazine. Reducing sugars were derivatized with DABS-hydrazide at 50 degrees C for 120 min. The chromophoric hydrazones were separated by reversed-phase HPLC isocratically using a short column (4.6 X 50 mm) and 0.08 M acetic acid-acetone as an eluant with no sample pretreatment and were quantitated at the picomole level. This method was applied to the sugar analysis of 5 micrograms of glycoproteins. Dansyl hydrazide derivatives of sugars were also separated by this HPLC system.

Carbohydrates↗

Amino acid sequence of two sea anemone toxins from Anthopleura fuscoviridis.

The amino acid sequences of two toxins, AFT-I and AFT-II, from the sea anemone Anthopleura fuscoviridis were determined. AFT-I and -II consist of 47 and 48 amino acid residues, respectively, and are cross-linked with three disulfide bridges. The sequences have high homology to those of toxins I and II from Anemonia sulcata and anthopleurin-A and -B from Anthopleura xanthogrammica.

Amino Acid Sequence↗

Isolation and characterization of agglutinins from the hemolymph of an acorn barnacle, Megabalanus volcano.

Two agglutinins, MVA-1 and MVA-2, were isolated from the hemolymph of the acorn barnacle, Megabalanus volcano. They agglutinated human erythrocytes irrespective of the ABO blood group and also rabbit and sheep blood cells. Lactose and fetuin strongly inhibited the hemagglutinating activity. D-galactose, D-arabinose and N-acetylneuraminic acid were also moderate inhibitors. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, both MVA-1 and MVA-2 gave a single band corresponding to 38,000 daltons. It split into one major band with a molecular weight of 23,000 in the presence of 2-mercaptoethanol. The two agglutinins showed the same apparent molecular weight of 116,000 by gel filtration. In isoelectric focusing MVA-1 showed one band at pH 4.8, whereas MVA-2 gave a main band at pH 4.4 with few faint ones in the range between pH 4.0 and 4.8. The agglutinins were glycoproteins containing D-mannose and L-fucose as carbohydrate components. No precipitation reaction was observed in Ouchterlony immuno-diffusion tests using rabbit antisera against the agglutinins from the phylogenetically related Megabalanus rosa.

Amino Acids↗

A microcomputer aided control system for visual tracking task of monkey.

A microcomputer aided control system for visual tracking tasks is described. A video monitor displays positions of target and pointer. In a handle task, the horizontal position of a pointer is coupled to the angular position of a manipulandum. The task involves alignment and maintenance of a pointer on the target. In a bar press task, the pointer moves incrementally after each preset number of bar presses. A microcomputer and video monitor afford system flexibility, while a panel of manual switches permits the setting up of a variety of experimental conditions without extensive keyboard inputs and has the added advantage of constant display of task details. The program is written in assembly language to meet requirements for program control speed and allow timing measurement in msec. This system may be useful in the behavioral study of physiological and neurophysiological functions.

Animals↗

Aplysianin-A, an antibacterial and antineoplastic glycoprotein in the albumen gland of a sea hare, Aplysia kurodai.

Aplysianin-A, an antibacterial and antineoplastic factor in the albumen gland of the sea hare Aplysia kurodai, was isolated. It had a molecular weight of approximately 320 kD and consisted of subunits with a molecular weight of 85 kD. It contained 9.8% neutral sugar. Aplysianin A showed 50% inhibition of Bacillus subtilis growth at a concentration of 4 microgram protein/ml and 50% lysis of murine MM46 tumor cells at 14 ng protein/ml. A partial identity of antigenic specificity of the purified specimen with an antineoplastic factor from Aplysia eggs was observed in immunodiffusion tests.

Albumins↗

Eosinophil chemotactic factors from granuloma of Kimura's disease, with special reference to T lymphocyte-derived factors.

The granuloma of patients with Kimura's disease characterized by tissue and peripheral blood eosinophilia was reviewed with respect to eosinophil infiltration. An infiltrate of inflammatory cells with histiocytes and a sprinkling of eosinophils were observed in the fibrous stroma surrounding the newly formed vessels. Mast cells were rarely seen in the areas where eosinophils were grouped together. Three different eosinophil chemotactic factors (ECF) were isolated from the granulomas of Kimura's disease. They were termed as low molecular weight (LMW), intermediate molecular weight (IMW), and high molecular weight (HMW)-ECF according to the profile on gel filtration (LMW-ECF, about 500; IMW-ECF, about 12,500; HMW-ECF, 45,000-70,000). In terms of their activity when extracted from the granuloma, LMW-ECF and HMW-ECF seemed to be major natural mediators for the tissue eosinophilia, whereas IMW-ECF was a minor one. In an in vitro system, it was shown that granuloma lymphoid cells produce spontaneously at least two ECF having similar properties to LMW- and HMW-ECF, respectively. By analysis with monoclonal antibodies, granuloma T cells, probably OKT4-positive cells, were shown to be responsible for the production of those two ECF. It was thus suggested that prolonged synthesis of LMW- and HMW-ECF by OKT4-positive T cells plays a crucial role in the local eosinophilia of Kimura's disease.

Angiolymphoid Hyperplasia with Eosinophilia↗

A microcomputer-based system for detailed analysis of rat behavior.

A microcomputer-based system that can measure rotational behavior and moving velocity is described. Infrared photobeams detect the animal behavior. One advantage of this system is that data can be stored on floppy disks and analyzed in arbitrary time intervals. This system may be useful for detailed analysis of rat behavior in physiological or psychopharmacological experiments.

Animals↗

Isolation and characterization of hemagglutinins from the sponge Dysidea herbacea.

The sponge Dysidea herbacea (Keller) was found to possess hemagglutinins. The major component, DHA-I, is a protein with a mol. wt of 26,000, which dissociates into subunits of equal size (14,000). It contains large amounts of glutamic acid and aspartic acid residues, but no half-cystine, methionine or histidine residues. DHA-I reacted with rabbit and human AB0 erythrocytes. D-galactose and lactose were effective inhibitors of DHA-I. The sponge also contained a minor component(s) which reacted preferentially with rabbit erythrocytes but not with human AB0 erythrocytes.

Amino Acids↗

Movement and non-movement related pallidal unit activity during bar press feeding behavior in the monkey.

Activity was recorded from 358 neurons in the globus pallidus (GP) of monkeys (Macaca fuscata) during an operant feeding task consisting of 3 stages: (1) food or non-food presentation (1st stage); (2) bar pressing (2nd stage); and (3) food acquisition and ingestion (3rd stage). There were two kinds of neurons, one with high and the other with very low (almost silent), spontaneous firing rates. Two hundred and four neurons (57%) responded in one or more of the feeding stages. Of the 21 neurons which responded in the 1st stage, two responded selectively to food presentation, and 19 responded to both food and non-food visual presentation. One hundred and seventy-four neurons (49%) and 107 neurons (30%) responded in the 2nd and 3rd stages, respectively, and 106 (30%) of these were directly related to specific feeding motor acts such as arm extension, flexion, bar pressing, grasping, chewing etc. Both high and low firing neurons responded to motor acts with sharp or gradual onset. More than half of those that responded to arm extension showed laterality (contra or ipsi)- and function (extension or flexion)-dependent responses. The incidence of the motor related neurons was higher in the caudodorsal part of the GP. On the other hand, about one third, especially in the rostroventral part of the GP, showed dissociating responses in that they responded during bar pressing for food or during ingestion in an operant task, but not during bar pressing for non-food or during forcible ingestion. The magnitude of firing changes during arm extension and bar pressing depended on the nature of the food. Moreover, in trials using new food or false (model) food, firing changes during bar press appeared or disappeared within a few trials with no correlation to bar press movement. These data suggest heterogeneous functions within the GP; the caudodorsal part is strictly concerned with motor execution and preparation, while the rostroventral part is not related to motor function directly, but may rather be important in coupling internal, motivational information to the motor system.

Animals↗

The effects of feeding and rewarding brain stimulation on lateral hypothalamic unit activity in freely moving rats.

In some medial forebrain bundle (MFB) sites, self-stimulation is often modulated by hunger or satiety. With electrodes in the nucleus accumbens (NAC) such modulation rarely occurs. The influence of food deprivation on MFB self-stimulation is the main basis for the hypothesis that electrical stimulation of the MFB can mimic the rewarding effect of food for hungry animals. To investigate this hypothesis, unit activity was recorded from the lateral hypothalamic area (LHA) of freely moving rats during rewarding stimulation at loci in both MFB and NAC, and during food ingestion. Of 63 neurons tested during MFB stimulation, 41 were inhibited, 19 were activated, and 3 were not influenced. NAC stimulation suppressed 8 of the 31 neurons tested, excited 16, and elicited no response in the remaining 7. During ingestion, 29 of the 63 neurons tested were inhibited and one was facilitated. Of 29 neurons suppressed by food, 20 were also inhibited by rewarding MFB stimulation, but 10 of 13 neurons inhibited by food were excited by rewarding NAC stimulation. Thus, most LHA neurons inhibited during feeding were also inhibited by rewarding MFB stimulation. Rewarding NAC stimulation, however, does not inhibit most LHA neurons that are inhibited by food. This result suggests that LHA neurons which are inhibited by food might be involved in mediation of the rewarding effect of electrical stimulation at some sites in the MFB. Nevertheless, self-stimulation may occur by activating reward processes other than those related to food, because rewarding NAC stimulation does not inhibit LHA neurons which are suppressed by food.

Action Potentials↗

The application of fluorescein isothiocyanate and high-performance liquid chromatography for the microsequencing of proteins and peptides.

Amino acid derivatives of fluorescein isothiocyanate have been separated by reverse-phase high-performance liquid chromatography in 45 min, using a linear gradient formed from acetone and 10 mM sodium phosphate buffer (pH 7.0) at 60 degrees C. The fluorescence of the derivatives has been used for detection giving a sensitivity of less than 0.5 pmol for a single component. Applications of this method to the sequence analyses of egg white lysozyme and salmon beta-melanotropin are also described.

Amino Acid Sequence↗

Caudate unit activity during operant feeding behavior in monkeys and modulation by cooling prefrontal cortex.

Activity was recorded from 351 neurons in the head of the caudate nucleus (CD) of monkeys during an operant feeding task consisting of: (1) food or non-food presentation (P); (2) bar pressing (B); and (3) food acquisition and ingestion (I). Of 45 neurons which responded in the P phase and were tested systematically, 27 responded to visual presentation of both food and non-food (non-specific response), and 18 responded to food presentation only (food specific response). The magnitude of food specific responses depended on the nature of the food and was inversely related to the latency of the onset of bar pressing. Thirty-five neurons responded in the B phase: 28 changed firing rate continuously with no correlation to individual bar presses, while the activity of the other 7 was related to each bar press. In the I phase, 62 neurons responded to separate events: the activity of more than half (39 neurons) was often related to chewing movement or gustatory stimuli, and that of one third (23 neurons) changed during individual arm movements. The neurons which responded in the P phase were found to be distributed widely in the head of the CD except for its central zone, while the neurons which responded in the I phase were in the medial part. Cooling of the dorsolateral prefrontal cortex abolished the continuous responses seen in the B phase, but did not abolish the feeding behavior. The data suggest that in the head of the CD there are several groups of neurons that have different functions and different distributions: food specific, sensory integration responses, non-motor responses driven by the frontal cortex, motor responses coupled to various movements, and sensory responses which apparently originate in the intra-oral cavity. These functions may arise sequentially, or in correspondence with integration of the sensory and motor systems to produce coordinated behavior.

Animals↗

Preparation of a specifically tritiated locust adipokinetic hormone analog with full biological potency.

A synthetic peptide related to locus adipokinetic hormone ( AKH ) and shrimp red pigment concentrating hormone ( RPCH ) containing a tyrosine residue in place of phenylalanine was iodinated and the 3,5- diiodotyrosyl derivative was isolated by reverse phase HPLC. Catalytic dehalogenation of the diiodo derivative in the presence of tritium yielded the tritiated AKH analog which was isolated by gel filtration on Sephadex LH-20 and reverse phase HPLC. The tritiated peptide was formed to be identical to AKH in its ability to stimulate lipid release into the hemolymph of locusts in vivo where the diiodotryrosyl derivative was inactive. The specific radioactivity of the tritiated peptide was 57.2 Ci/mmol, or 99% of the theoretical value.

Animals↗