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Biomedical subjects

K Aihara

Publications and source records attributed to K Aihara.

At least 19 recordsLinked to original sources

Pruning of rat cortical taste neurons by an artificial neural network model.

1. Taste qualities are believed to be coded in the activity of ensembles of taste neurons. However, it is not clear whether all neurons are equally responsible for coding. To clarify the point, the relative contribution of each taste neuron to coding needs to be assessed. 2. We constructed simple three-layer neural networks with input units representing cortical taste neurons of the rat. The networks were trained by the back-propagation learning algorithm to classify the neural response patterns to the basic taste stimuli (sucrose, HCl, quinine hydrochloride, and NaCl). The networks had four output units representing the basic taste qualities, the values of which provide a measure for similarity of test stimuli (salts, tartaric acid, and umami substances) to the basic taste stimuli. 3. Trained networks discriminated the response patterns to the test stimuli in a plausible manner in light of previous physiological and psychological experiments. Profiles of output values of the networks paralleled those of across-neuron correlations with respect to the highest or second-highest values in the profiles. 4. We evaluated relative contributions of input units to the taste discrimination of the network by examining their significance Sj, which is defined as the sum of the absolute values of the connection weights from the jth input unit to the hidden layer. When the input units with weaker connection weights (e.g., 15 of 39 input units) were "pruned" from the trained network, the ability of the network to discriminate the basic taste qualities as well as other test stimuli was not greatly affected. On the other hand, the taste discrimination of the network progressively deteriorated much more rapidly with pruning of input units with stronger connection weights. 5. These results suggest that cortical taste neurons differentially contribute to the coding of taste qualities. The pruning technique may enable the evaluation of a given taste neuron in terms of its relative contribution to the coding, with Sj providing a quantitative measure for such evaluation.

Animals

Synthetic study of 2-[(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9- yl)-sulfinyl]-1H-benzimidazole analogs and their biological properties as novel proton pump inhibitors.

A series of 2-[(cycloalka[b]pyridinyl)sulfinyl]-1H-benzimidazoles (11) was synthesized and tested for antisecretory activity against pentagastrin-induced gastric acid secretion in rats. A novel benzimidazole derivative containing a cyclohepta[b]pyridine moiety was found to be the most potent among the congeners, which included five- to eight-membered cycloalka[b]pyridine ring systems. Some 2-[(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)sulfinyl]-1H- benzimidazole analogs (14) with various substituents on the aromatic rings showed superior properties to omeprazole (1) in biological examinations in vivo. A diastereoisomer, TY-11345 (28Ba), was selected as a promising agent for further evaluation.

Animals

Electron microscopical studies of the cell population in nasal secretions.

The purpose of this study is to identify the cell types and ultrastructural changes of the cells in nasal secretions, and to understand the pathology of allergic and infectious rhinitis. Nasal secretions from 20 patients with allergic rhinitis and 15 patients with infectious rhinitis have been observed by transmission electron microscopy. The cell population of the allergic group consists of (in order of predominance): epithelial cells, eosinophils, neutrophils, lymphocytes, basophilic cells (basophil leukocytes and mast cells), and macrophages. In the infectious group the population contains: neutrophils, epithelial cells, macrophages, and lymphocytes. Marked degranulation has been observed in the granules of eosinophils in allergic nasal secretions together with granule fusion, vacuolation, and signs of phagocytosis. Increased numbers of basophil leukocytes and mast cells are also a feature of the allergic nasal secretion. Degranulation of neutrophils is markedly increased in the infectious group as compared to the allergic group. Clustered epithelial cells are observed in the allergic group more often than in the infectious group. Four types of lymphocytes with different morphological features are observed in both groups, i.e. small lymphocyte, T-lymphocyte-like cells, large granular lymphocyte-like cells, and plasma cells. The results of the present study show special ultrastructural characteristics in the cell population of allergic nasal secretions, i.e., an increase in the number of degranulated eosinophils and basophilic cells, clustered epithelial cells, and large granular lymphocytes, while an increase in degranulated neutrophils and macrophages with marked phagocytosis are characteristic for infectious nasal secretions.

Adult

Nizofenone, a neuroprotective drug, suppresses glutamate release and lactate accumulation.

It was investigated whether nizofenone, 2'-chloro-2-(2-diethylaminomethyl-1- imidazolyl)-5-nitrobenzophenone fumarate, shows neuroprotective effect by inhibiting glutamate release and lactate accumulation in rats. Nizofenone (10 mg/kg i.p.) significantly inhibited neuronal cell death in the hippocampus CA1 pyramidal cells 7 days after 15 min of ischemia caused by 4-vessel occlusion. The extracellular level of glutamate and lactate increased in the hippocampus during ischemia. Upon reperfusion, the glutamate level decreased rapidly but, in contrast, the lactate level increased further and reached a maximum at 15 min following reperfusion. Nizofenone (10 mg/kg i.p.) completely inhibited the ischemic glutamate increase and post-ischemic lactate increase. These results suggest that the inhibition of ischemic release of glutamate and post-ischemic accumulation of lactate could contribute greatly to the neuroprotective effect of nizofenone.

Analysis of Variance

Purification and characterization of recombinant human macrophage colony-stimulating factor expressed in Chinese hamster ovary cells.

We expressed human macrophage colony-stimulating factor (M-CSF) in Chinese hamster ovary (CHO) cells by introducing an expression plasmid coding for a 554-amino-acid M-CSF precursor and dihydrofolate reductase (DHFR) gene, and by amplifying the sequence. A cell line was obtained that secreted approximately 200,000 units/ml after 6 days in culture. The expressed recombinant human M-CSF (rhM-CSF) primarily consisted of two molecular species, a main 80-90 kD M-CSF as a homodimer and a molecular form higher than 150 kD. Purification of a main rhM-CSF gave an apparently homogeneous protein disulfide-bonded from 42-kD subunits, but one of the purified rhM-CSFs was composed of two subunit species with molecular masses of 44 and 42 kD. These purified rhM-CSFs had substantially the same specific activity (1 to 4 x 10(7) units/mg protein). Deglycosylation experiments with the latter rhM-CSF using chemical (trifluoromethanesulfonic acid) and enzymatic methods found a terminal neuraminic acid in addition to N- and O-glycosylation, but the two subunit species did not coalesce into a single molecule.

Amino Acid Sequence

Effects of Y-20811, a thromboxane A2 synthetase inhibitor, on experimentally induced coronary thrombosis in anesthetized dogs.

The effects of Y-20811, a selective inhibitor of thromboxane A2 (TXA2) synthetase, on blood flow and local levels of immunoreactive thromboxane B2 (i-TXB2) and 6-keto prostaglandin F1 alpha (i-6-keto PGF1 alpha) in the coronary artery were investigated in the canine model of coronary thrombosis. Thrombosis was induced by applying an electric current to the intraluminal surface of the coronary artery. The plasma levels of i-TXB2 and i-6-keto PGF1 alpha were measured distal to the electrode. In the control group, coronary blood flow decreased and finally stopped 207 +/- 53 min (mean +/- S.E.M., n = 5) after the start of current application. The level of i-TXB2 rose before the coronary occlusion. Coronary blood flow did not change significantly in the Y-20811-treated group (1 mg/kg i.v.). The level of i-TXB2 decreased and remained significantly lower than that in the control group. The level of i-6-keto PGF1 alpha tended to increase slightly in the Y-20811-treated, but not in the control group. The weight of the thrombus in the Y-20811-treated group was significantly less than that in the control group (P less than 0.01). These results suggest that Y-20811 prevents coronary thrombosis by the inhibition of TXA2 production around the electrically injured lumen of the coronary artery.

6-Ketoprostaglandin F1 alpha

A novel method to analyse response patterns of taste neurons by artificial neural networks.

Despite several notions on the gustatory code proposed over three decades, investigators have not yet reached a consensus. This paper describes a new approach to analyse gustatory neural activities. Three-layer neural networks were trained by the back-propagation learning algorithm, to classify the neural response patterns to four basic taste qualities. The discrimination by the trained networks on taste qualities in the response patterns of rat chorda tympani fibres (CT) and cortical taste neurons (CN) was consistent both with the correlation analysis and with behavioural experiments. By examining the connection weights of each neuron, some input neurons representing CN were 'pruned' without deteriorating the ability of the network to discriminate taste. This characteristic of the network is contrary to a previous hypothesis, that taste neurons are of equal importance in the neural coding.

Animals

The antihypertensive effect of 2-alkynyladenosines and their selective affinity for adenosine A2 receptors.

We examined the affinity for adenosine receptors and the antihypertensive effects of 2-alkynyladenosines, especially 2-hexynyladenosine (2-H-Ado) and 2-octynyladenosine (2-O-Ado). The order of decreasing affinity of 2-H-Ado, 2-O-Ado, and other agonists tested for A1 receptors was N6-cyclopentyladenosine (CPA) greater than N6-cyclohexyladenosine (CHA) greater than N6-R-phenylisopropyladenosine (R-PIA) greater than 2-chloroadenosine (CADO) = 5'-N-ethylcarboxamideadenosine (NECA) greater than N6-S-phenylisopropyladenosine (S-PIA) greater than 2-H-Ado greater than 2-O-Ado greater than 2-phenylaminoadenosine (CV-1808), and that for A2 receptors was 2-H-Ado greater than 2-O-Ado = NECA greater than CADO greater than CV-1808 greater than R-PIA greater than CPA greater than CHA greater than S-PIA. The Ki values of 2-H-Ado and 2-O-Ado for [3H] NECA binding to A2 receptors were 4.1 and 12.1 nM, respectively, and those for [3H]CHA binding to A1 receptors were 146 and 211 nM, respectively: the affinity of 2-H-Ado and 2-O-Ado for A2 receptors was about 36- and 17-fold higher than their affinity for A1 receptors. Injection of 2-H-Ado and 2-O-Ado (0.03-100 micrograms/kg) decreased the blood pressure of anaesthetized spontaneously hypertensive rats (SHR). A slight decrease in heart rate was observed after i.v. injection of 100 micrograms/kg 2-H-Ado and 2-O-Ado. A potent and long-lasting antihypertensive effect was also observed after oral administration of 2-H-Ado and 2-O-Ado to conscious SHR.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine

Relations between contractile responses and beta-adrenoceptors in gastric fundus of diabetic rats.

Contractile responses to norepinephrine (NE), and the population of beta-adrenoceptors, were determined in gastric fundus smooth muscle from rats with diabetes induced by streptozotocin (STZ), and age-matched controls. Relaxation and/or contraction of fundus strips of controls and diabetics were induced by 10(-5)M NE. Responses to NE were mainly relaxation in gastric fundus isolated from controls, and contraction in fundus isolated from diabetics. Contraction was blocked by 10(-8) M prazosin and relaxation was blocked by 10(-6) M propranolol. Relaxation by isoproterenol of contraction induced by 10(-6) M acetylcholine was significantly less in fundus from diabetics than in that from controls. The number of beta-adrenoceptors, measured with [125I] iodocyanopindolol as a ligand, was significantly less in gastric fundus membrane isolated from diabetics than in that from controls, but affinity was no different. The level of plasma catecholamine was higher in diabetics than in controls. Results suggest that depression of gastric fundus relaxation and increase of contraction by NE in diabetics could be due to fewer beta-adrenoceptor binding sites caused by down-regulation by higher catecholamine level in diabetic rats.

Animals

Experimental studies of moderate temperature burns.

The pathology and characteristics of moderate temperature burns have been investigated in rats with a heating apparatus capable of applying thermal doses varying in temperature and exposure time, to which the responses have been observed both macroscopically and microscopically. The macroscopic examination showed only erythema and greyish necrosis. The microscopic changes were divided into six grades of severity from 0 to 5. From the time-temperature threshold curves made by microscopic observations, the critical temperatures for superficial and deep dermal burns, and full skin thickness burns were 37.8 degrees C, 41.9 degrees C and 47.9 degrees C, respectively. According to the data, almost all commercially available topical heaters are considered hazardous. Susceptibility to thermal injury was markedly increased by compression and ischaemia, the effects of which are thought to be due mainly to rapid heat accumulation (heat damage) and hypoxaemia (hypoxic damage).

Animals

Detection of heat-stable delta 3,delta 2-enoyl-CoA isomerase in rat liver mitochondria and peroxisomes by immunochemical study using specific antibody.

The subcellular distribution of delta 3,delta 2-enoyl-CoA isomerase [EC 5.3.3.8] and the inducing effect of clofibrate, a peroxisomal proliferator, on the enzyme activity were examined in rat liver. From the results of spectrophotometric investigation of the fractions, which were prepared by sucrose discontinuous gradient centrifugation from the light mitochondrial fraction, the isomerase activity was found in the fractions enriched in mitochondria and those enriched in peroxisomes of the control and the clofibrate treated rat livers. The anti-isomerase antibody reacted with both the mitochondrial isomerase and the peroxisomal isomerase, revealing a single band with an apparent molecular weight of 30,000. However, the isomerase was induced by clofibrate administration mainly in the mitochondrial fraction. These results suggest that delta 3,delta 2-enoyl-CoA isomerase is located in the mitochondria and the peroxisomes of the normal rat liver, and that the isomerase in the mitochondria is induced by clofibrate administration.

Animals

Silver staining of nucleolar organizer regions in prostatic lesions.

Variations in the number of silver-stained nucleolar organizer region-associated proteins (AgNORs) were studied in paraffin sections of 42 benign prostatic lesions, comprising four cases of granulomatous prostatitis, five of squamous or transitional metaplasia, eight of atypical and 25 of regular hyperplasia, and 37 of prostatic adenocarcinoma, with their metastases. There was a significant difference between the mean AgNOR counts of the benign and malignant prostatic lesions (1.58 +/- 0.26 v. 4.34 +/- 1.53; P less than 0.01). The mean AgNOR counts significantly increased with increasing Gleason's grade (P less than 0.01) and clinical stage (P less than 0.05) of the tumours. AgNOR counting may contribute to the conventional diagnostic and prognostic indices of cancer of the prostate.

Adenocarcinoma

A histological, ultrastructural and immunohistochemical study of superficial temporal arteries and middle meningeal arteries in moyamoya disease.

Pathologic changes in superficial temporal arteries (STA) and middle meningeal arteries (MMA) biopsied from 15 patients with moyamoya disease (MD) who had undergone cerebro-temporal arterio-synangiosis were studied histologically, ultrastructurally and immunohistochemically. The main pathologic features were: proliferation of smooth muscle cells (SMCs) and thickening of the intima, degeneration and destruction of SMCs in the media and intima, and the presence of condensed organelles in necrosed SMCs or the interstitium among SMCs, or both outside and within the elastica interna (EI). The EI had become thin, porous, fragmented and was even absent in some segments. These changes are different from those of other forms of angiopathy, but identical with those at the ends of internal carotid arteries (ICA) reported by us previously, being pathognomonic for MD. These changes in the STA and MMA reveal that MD involves not only the ICA but also the intra- and extracranial branches of external carotid arteries. The medial necrosis of SMCs seems to be the primary injury of the arterial wall in MD. STA tissue blocks from two cases of MD were stained immunohistochemically. By electron microscopy, IgG-, IgM-, and C3-positive granules were observed on the ER of endothelial and intimal cells. Further studies on more cases are needed to determine whether an immunoreaction has occurred in these arteries.

Adolescent

Pharmacological profile of the 2-alkynyladenosine derivative 2-octynyladenosine (YT-146) in the cardiovascular system.

We investigated the cardiovascular effects of 2-octynyladenosine (YT-146), an adenosine A2 agonist, in various mammalian preparations in comparison with adenosine and 2-chloroadenosine. YT-146, when intravenously administered, caused a dose-dependent decrease of blood pressure in anesthetized normotensive rats (with ED30 values of 0.4 micrograms/kg), and YT-146 was 250 times more potent than adenosine. Whereas adenosine and 2-chloroadenosine decreased heart rate at approximately equihypotensive doses, YT-146 had no negative chronotropic effects at h hypotensive doses. Orally given YT-146 (0.1 - 1 mg/kg) produced a potent and long-lasting antihypertensive effect in spontaneously hypertensive rats. YT-146 was 15.9 and 12.5 times more potent than adenosine in producing relaxation of isolated porcine coronary arteries and in increasing dog coronary blood flow, respectively. Although YT-146 was equipotent to adenosine in causing a negative inotropic effect in isolated guinea pig atria, it was less potent than adenosine in producing atrioventricular conduction block in guinea pigs. On the other hand, 2-chloroadenosine was 9.1, 1.8 and 2.4 times more potent than adenosine in lowering blood pressure, relaxing isolated porcine coronary arteries and increasing dog coronary blood flow, respectively. 2-Chloroadenosine was the most potent in producing cardiodepression, i.e., negative inotropy and atrioventricular conduction block in guinea pigs. From these results, we concluded that YT-146 is a potent coronary vasodilator and also a potent, orally active and long-acting hypotensive agent having less cardiac depressant activity.

Adenosine

Nitrendipine facilitates recovery of cerebral blood flow, EEG and metabolites following cerebral ischemia in anesthetized rabbits.

The effect of nitrendipine, an antihypertensive calcium antagonist, on the impairment of cerebral blood flow and EEG observed after 10-min complete cerebral ischemia in anesthetized rabbits was compared with those of nicardipine. The ischemia was produced by neck tourniquet in combination with hypotension (50-60 mmHg). Blood flow was measured by hydrogen-clearance method. Transient reactive hyperemia was observed immediately after the cessation of ischemic procedure, and was followed by a decrease in blood flow in the range of 58-73% of corresponding basal values in the total brain, cortex and thalamus. The postischemic decrease in blood flow was suppressed when nitrendipine (0.3-1 mg/kg) or nicardipine (3-10 mg/kg) was given intraduodenally before ischemia. The postischemic decrease in total intensity and frequency index in EEG recovered rapidly when nitrendipine was pretreated, whereas the recovery of EEG parameters was not obtained by the nicardipine pretreatment. These results suggest that the effect of nitrendipine and nicardipine on the postischemic cerebral blood flow may be due to the inhibition of calcium-induced contraction in cerebral vessels, whereas the discrepancy between the effects of these agents on EEG may not be due solely to the improvement in cerebral circulation. Furthermore, the improvement in postischemic cerebral energy metabolism was confirmed by nitrendipine pretreatment (0.3 mg/kg).

Animals

Comparison of the protective effects of the pertussis acellular vaccine with the component vaccine, which have different amounts of fimbriae, against the experimental aerosol infection of mice with Bordetella pertussis.

We obtained highly purified fimbrae from Bordetella pertussis cells which gave a single band in SDS-polyacrylamide gel electrophoresis. Using the purified fimbriae, we prepared anti-fimbriae rabbit or goat IgGs by fimbriae-Sepharose affinity chromatography, and developed ELISA for its determination. The concentration of fimbriae in our former type of acellular vaccine (80 micrograms per ml) was determined to be 20 ng per ml, and that in our component vaccine was at an almost negligible level. To estimate the protective effects of the small levels of fimbriae, we evaluated the protective activities of three types of vaccines; the former type acellular vaccine (pertussis toxin (PT): filamentous hemagglutinin (FHA): fimbriae = 15:61:0.02 (microgram per ml)), the two-component vaccine (16:64:0.0001), and the three-component vaccine (16:63:0.8). The three types of vaccines showed no significant differences in protectivity against the experimental aerosol infection suggesting that these levels of fimbriae are not effective against the experimental aerosol infection of mice with Bordetella pertussis.

Adhesins, Bacterial

Application of cryo-sapphire knife in rapid frozen ultrathin sectioning for electron microscopy.

Conventional sectioning of frozen and fixed tissues is usually performed using the glass knife. Although the glass knife is economical, it has several disadvantages. In this study, the Authors have examined the sectioning quality of the sapphire knife in frozen tissues as an alternative to the glass knife. It was concluded that the sapphire knife has better quality and durability than the conventional glass knife in cutting frozen tissues.

Animals

Further evaluation of the pertussis component vaccine produced by apoceruloplasmin affinity chromatography.

The heat-treated apoceruloplasmin (Apocp) is a useful protein as an affinity ligand for the purification of pertussis toxin (PT). The amounts of Apocp in the purified antigens or the pertussis component vaccine were determined. Anti-Apocp antibodies were not detected by the passive cutaneous anaphylaxis (PCA) test in rats. No anti-Apocp antibody was detected after hyperimmunization of rabbits with the vaccine. Apocp was not detected in PT and filamentous hemagglutinin (FHA) by ELISA using rabbit anti-Apocp IgG. In the experiments using 125I-labelled Apocp, 125I-Apocp was not detected in either PT or FHA which were purified by 125I-labeled Apocp-Sepharose, DEAE Sepharose, and cellulose sulfate chromatography. The contents of human DNA were also determined to be less than 10 pg per 1 mg of Apocp, by the dot-blot hybridization method using the 32P-labeled DNA probe of Alu sequence. In the tests for the presence of inapparent viruses, HBs antigen and HTLV-III antibody, no contamination was found in either the Apocp or in the vaccine. Large amounts of various viruses, which were intentionally added to the Apocp (spiking test), were completely inactivated by heating at 65 degrees C for 18 hr. Both the Apocp and the vaccine passed the general pharmacology and acute toxicity tests. From these results, the heat-treated Apocp was considered to be a suitable affinity ligand for the purification of the antigens for the pertussis component vaccine.

Adhesins, Bacterial