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Biomedical subjects

K Kurihara

Publications and source records attributed to K Kurihara.

At least 91 records · Page 5Linked to original sources

Detection of triazolam in skeletal remains buried for 4 years.

Analyses of the hypnotic triazolam from the remains of two human skeletons buried underground for 4 years were made for purposes of confirmation. The bone marrow and mummified muscle were digested with 2 M sodium hydroxide, efficiently extracted using a 3-step solvent extraction procedure, and selectively analyzed by gas chromatography/mass spectrometry with the negative ion chemical ionization mode. Estazolam was the internal standard used. Triazolam was detected in all the samples; the concentrations were 0.36 ng/g in the bone marrow of one victim, and 0.37 and 5.5 ng/g in the bone marrow and mummified muscle of the other victim. This method should prove useful for determination of triazolam in extensively decomposed bodies.

Bone Marrow↗

Distribution of neuropeptide-containing nerve fibers in the human submandibular gland, with special reference to the difference between serous and mucous acini.

Distribution of neuropeptide Y (NPY)-, vasoactive intestinal polypeptide (VIP)-, galanin (GAL)-, substance P (SP)-, and calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibers in the human submandibular gland was examined by the peroxidase-antiperoxidase method with attention to high-quality fixation and the condition of patients. NPY-, VIP-, and GAL-immunoreactive varicose fibers were densely distributed around the acini and ducts. Some of these fibers extended between acinar cells. The density of SP- and CGRP-immunoreactive fibers was relatively low. The number of NPY-, VIP-, and GAL-immunoreactive fibers around the mucous acini was significantly higher than around the serous acini. In the perivasculature, NPY-immunoreactive fibers were more numerous than other immunoreactive fibers. No somatostatin-, leucine-, or methionine-enkephalin-immunoreactive fibers were detected. Our findings suggest that a large number of periacinar VIP-, NPY-, and GAL-immunoreactive fibers may participate in regulating the synthesis of saliva and its secretion. Since the VIP-, NPY-, and GAL-immunoreactive fibers are more numerous around the mucous acini than around the serous ones, these fibers may take part more actively in regulating the secretory mechanisms in the mucous acini than in the serous ones. The relatively low number of CGRP- and SP-immunoreactive fibers suggests that they are less involved in the function of the human submandibular gland. Perivascular peptidergic fibers, especially NPY-immunoreactive fibers, may be involved in controlling local blood flow in this gland.

Aged↗

A calcium channel in human submandibular duct cell line, HSG cells, not regulated by P2U purinergic receptor-mediated intracellular calcium mobilization.

Signal transduction via P2 purinergic receptors was investigated in HSG cells, a continuous cell line originally derived from an irradiated human salivary gland. Ligand specificity for nucleotide receptors in HSG cells was investigated with various nucleotides and their analogues. Inositol 1,4,5-trisphosphate (IP3) production was significantly increased by ATP, UTP and ATP gamma S. The ligand specificity of this effect agreed well with that of the P2U purinergic receptor. On the other hand, 45Ca2+ influx was stimulated by ATP, UTP > ATP gamma S, ADP, UDP > ADP beta S > AMPPNP, GTP, TTP > CTP, GDP, TDP, AMPPCP, AMPCPP. This ligand specificity of 45Ca2+ influx was much broader than IP3 production. Also pertussis and cholera toxin had no effect on both IP3 production and 45Ca2+ influx by ATP or UTP. 3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate (Bz-ATP) stimulates 45Ca2+ influx more effectively than IP3 formation. A 53-kDa membrane protein was photolabelled with [alpha-32P]Bz-ATP. This 53-kDa protein is a putative P2 purinergic receptor. In particular, the labelling was inhibited by a ligand profile that corresponded to that for 45Ca2+ influx. These findings suggest that nucleotides stimulate 45Ca2+ influx and IP3 formation by separate pathways via pertussis and cholera toxin-insensitive G proteins. Thus, in HSG cells, IP3 formation is coupled to the P2U subclass, while 45Ca2+ influx is coupled to another subclass, such as P2X, that regulates calcium channels.

Adenine Nucleotides↗

Tissue-specific methylation occurs in the essential promoter element of the tyrosine hydroxylase gene.

Expression of tyrosine hydroxylase (TH) is regulated in a tissue-specific manner by multiple mechanisms. In catecholaminergic cells, the expression of TH-mRNA is up-regulated by forskolin (FK) and is suppressed by retinoic acid (RA). We have previously provided evidence that, in N-18 cells, the expression of TH-mRNA is suppressed by DNA methylation of the TH gene itself. In the present study, using a catecholaminergic cell line, N1E-115, we performed deletional and mutational analyses on the 5'-flanking region of the mouse TH gene. The results indicate that a cAMP response element (CRE) mediates constitutive transcription of the TH gene, as well as responsiveness to FK and RA. Using bisulfite sequencing methods, we analyzed the methylation status of the TH gene 5'-flanking region in various cell lines and rat tissues. We found that three cytosine residues in the domain surrounding the CRE of the TH gene promoter were specifically methylated in N-18 cells and TH non-expressing rat tissues. In contrast, these cytosines were undermethylated in TH expressing cell lines and tissues. The inverse correlation between the frequency of cytosine methylation at these specific sites and the levels of TH expression supports a role for DNA methylation in the regulation of tissue-specific gene expression.

Animals↗

An immortalized septal cell line that expresses trkA mRNA in response to basic fibroblast growth factor.

Cells dissociated from embryonic rat septum were transfected with SV40 large T antigen gene by the calcium phosphate precipitation method. One of the cloned lines (EG6) had a flat morphology and expressed nestin gene, a marker of neural stem cells. Upon stimulation with basic fibroblast growth factor (bFGF), morphological changes of EG6 cells were induced, e.g. shrinkage of cell body and extension of neuritic processes. Furthermore, bFGF induced expression of the high affinity NGF receptor (trkA) gene. EG6 cell line thus will be useful for studies of roles of bFGF in the developing CNS as well as for studies of the regulation of trkA gene expression.

Animals↗

Odor discrimination of 'cAMP-' and 'IP3-increasing' odorants at high temperature and at high NaCl concentration in turtle olfactory system.

The ability of the turtle olfactory system to discriminate between various cAMP- and IP3-increasing odorants at high temperature and at high NaCl concentration in the olfactory bulb was examined by the cross-adaptation technique. The degrees of discrimination in high [Na+] solution were similar to those in normal Ringer's solution, suggesting that selectivities of receptors coupled with cAMP- and IP3-dependent pathways are similar to those coupled with both cAMP- and IP3-independent pathways. The mean values of the degree of discrimination among the IP3-increasing odorants were higher than those among the cAMP-increasing odorants at high temperature and at high [Na+] concentration. The degrees of discrimination among the IP3-increasing odorants at 40 degrees C were greater than those at 25 degrees C, while those among the cAMP-increasing odorants at 40 degrees C were similar to those at 25 degrees C, suggesting that the features of the receptors of cAMP-increasing odorants are different from those which respond to IP3-increasing odorants.

Adaptation, Physiological↗

Long-term recording of olfactory and vomeronasal stimulant-induced waves from the turtle main olfactory bulb and accessory olfactory bulb.

Recording of stimulant-induced waves (bulbar responses) from the main olfactory bulb is a useful tool for measuring quantitative stable olfactory responses. There is a good relationship between the olfactory bulbar response, olfactory nerve response and electroolfactogram (EOG), suggesting that the bulbar response reflects events in receptor cells. The modern whole-cell recording technique offers direct information on olfactory transduction in single cells, but it requires long experimental periods and many animals. On the other hand, analysis of bulbar responses provides useful information and requires the use of few animals. For example, we found that cAMP-increasing and IP3-increasing odorants were not distinctly received by the turtle olfactory organ by measuring olfactory bulbar responses and analyzed with a multidimensional scaling from about 60 animals. However, to record similar odor responses from isolated turtle olfactory neurons, at least 200 animals would be necessary. Bulbar responses are recorded with electrodes implanted into or located on the main olfactory bulb. When electrodes are located on the olfactory bulb surface, it is possible to record stable responses over a period of 3 days. These methods were applied successfully to the accessory olfactory bulb. In this paper, we describe the protocols used for recording of the stimulant-induced waves from the main and accessory olfactory bulb.

Adaptation, Physiological↗

Specific inhibitor for bitter taste: inhibition of frog taste nerve responses and human taste sensation to bitter stimuli.

Among various taste stimuli, bitter substances are most abundant and their chemical structures are greatly diverse from each other. It has been known that there are multiple receptor sites and transduction mechanisms. Already in neonates, bitter stimuli elicit rejection responses, indicating strong negative hedonic tone. Bitter taste is decidedly unpleasant when the sensation is strong. The development of a method to mask bitterness has widely been required in pharmaceutical sciences and food sciences. To mask bitterness, a specific bitterness inhibitor would be most useful. Such an inhibitor would also be useful in elucidating the receptor mechanisms of bitter substances. No inhibitor has, however, been available. Recently we found that a lipoprotein, PA-LG made of phosphatidic acid (PA) and beta-lactoglobulin (beta-LG), selectively suppresses the taste responses to bitter substances. In this paper we describe the protocol used for inhibition of the frog taste (glossopharyngeal) nerve responses to bitter stimuli by the lipoprotein. The frog taste system is used because it is sensitive to various bitter substances and surgery of the animal for the electrophysiological recording is rather easy. We also describe the protocol used for inhibition of human taste sensation to bitter stimuli.

Animals↗

Basic studies for the practical use of bitterness inhibitors: selective inhibition of bitterness by phospholipids.

PURPOSE: We examined the effects of phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol (PI), and phosphatidic acid (PA) on human taste sensation to various substances. METHODS: The effects were evaluated psychophysically using paid volunteers. RESULTS: PA inhibited the bitterness of various substances dissolved in water without affecting sweetness, saltiness, and sourness, although its inhibitory activity was less than that of PA-LG. PI also showed inhibitory activity on bitterness, although its activity was less than PA. A soybean lecithin fraction containing high contents of PA and PI also demonstrated inhibitory activity on the bitterness of various substances. Both the incorporation of either PA or the lecithin fraction into granules containing quinine and the coating of the granules with PA or the fraction effectively inhibited the bitterness of quinine. CONCLUSIONS: The lecithin fraction is permitted for use as an additive to drugs and food and can be produced on an industrial scale. It is expected that the lecithin fraction will be used safely as a bitterness inhibitor for practical applications.

Humans↗

Heat-induced apoptosis and p53 in cultured mammalian cells.

Heat-induced apoptosis was studied in M10 and MOLT-4 cells by determining nuclear morphological changes, decrease in cell size, DNA degradation into fragments of about 30 kbp, and the appearance of internucleosomal DNA fragments (DNA ladders). Morphological changes in the nucleus were detected within 30 min after heat-treatment at 44 degrees C in M10 cells, but much later (> 5 h) in MOLT-4 cells. In M10 cells, 30 kbp-DNA fragments were observed even at the end of the heat-treatment and decreased 10 min later, while the DNA ladder increased at 10-30 min after heat treatment. DNA fragments of 30 kbp appeared in MOLT-4 cells at 1 h after the heat-treatment and apparently accumulated for up to 24 h. Heat-treatment increased p53 protein in MOLT-4 cells but not in M10 cells. Analysis of the DNA sequence of the p53 gene revealed that M10 cells have a heterozygous mutation in codon 173 of exon 5. These results suggest that apoptosis is induced by hyperthermia in a cell-line dependent manner, that the formation of 30 kbp-DNA fragments is a very early event in apoptosis, that DNA fragmentation into a DNA ladder occurs via the 30 kbp fragments, and that apoptosis in heat-treated M10 cells may be independent of p53.

Apoptosis↗

Inositol-1,4,5-trisphosphate induces responses in receptor neurons in rat vomeronasal sensory slices.

Using the whole-cell mode of the patch-clamp technique, we recorded action potentials, voltage-activated cationic currents and putative second messenger-activated currents in receptor neurons in the vomeronasal sensory epithelium of female rats. The resting membrane potential and input resistance were -45.5 +/- 2.5 mV (mean +/- SEM, n = 39) and 1.5 +/- 0.2 G omega (mean +/- SEM, n = 37). Current injection of 1-3 pA induced overshooting action potentials. The firing frequency increased with increasing current injections linearly from 1 to 10 pA and reached a plateau at 30 pA, suggesting that rat vomeronasal receptor neurons sensitively elicit action potentials in response to a small receptor potential. Under voltage clamp, voltage-dependent Na+ inward current, inward Ca2+ current, sustained outward K+ current and Ca-(2+)-activated K(+)-current were identified. Dialysis of D-inositol-1,4,5-trisphosphate (D-IP3) induced inward currents with an increase in membrane conductance in approximately 54% of the cells and inward current fluctuations in 15% of the cell. L-IP3 also induced inward currents and current fluctuations in 53 and 13% of the cells respectively. The mean amplitude of inward currents induced by 100 microM D-IP3 and L-IP3 were 84.6 +/- 14.0 pA (SEM, n = 82) and 66.1 +/- 9.4 pA (SEM, n = 100) respectively. The IP3-induced responses were blocked by elimination of Na+ and Ca2+ in the external solution or application of 10 microM ruthenium red. The present study suggested that IP3-mediated transduction pathways exist in rat vomeronasal receptor neurons.

Action Potentials↗

Concentration and membrane fluidity dependence of odor discrimination in the turtle olfactory system.

In the present study, we examined the concentration dependence of odor discrimination in turtle olfactory bulbar responses using the cross-adaptation technique. In the odorant pairs with diverse molecular structures, the degree of discrimination was unchanged or only slightly decreased with an increase in odorant concentrations, suggesting that odorants are well discriminated even at high concentrations. In the odorant pairs with closely related molecular structures, the degree of discrimination was decreased with an increase in odorant concentrations. An increase in the temperature of turtle olfactory epithelium also decreased the ability to discriminate these odorants. There was a good correlation between changes in the odor discriminating ability induced by an increase in odor concentrations and those induced by a temperature increase. The liposomes were made of lipids extracted from the turtle olfactory epithelia and changes of their membrane fluidity induced by adsorption of odorants were monitored with DPH. There was a good correlation between a decrease in odor discriminating ability and the membrane fluidity changes induced by odorants. We suggest that decreases in odor discriminating ability induced either by an increase in odor concentration or by a temperature increase are ultimately caused by changes in the membrane fluidity.

Animals↗

Pigmented variant of benign oncocytic lesion of the pharynx.

A case of pigmented variant of benign oncocytic lesions is reported. The lesion was incidentally found in the pharynx of a 69-year-old man, and the gross appearances were multiple, small, flat elevations with black discoloration. Microscopically, ducts of seromucinous glands were replaced by melanin-containing oncocytes and adjacent dendritic melanocytes. To date, such a pigmented variant of benign oncocytic lesion has not been reported before.

Aged↗

Tophaceous pseudogout of the temporomandibular joint: report of a case.

An 85-year-old man was found to have a calcified mass protruding from the joint space of the right temporomandibular joint (TMJ). Microscopically, the removed mass consisted of chondromyxoid tissue with atypical chondrocytes, resembling a cartilaginous tumor. However, the chondromyxoid tissue contained abundant deposits of rod-shaped to rhomboid crystals which proved to be calcium pyrophosphate dihydrate (CPPD) crystals. The review of the literature revealed that tophaceous pseudogout was the most common variant of CPPD deposition disease involving the TMJ.

Aged↗

Sudden cardiac death in Japanese people aged 20-60 years--an autopsy study of 133 cases.

In order to elucidate the principal cause and actual circumstances of sudden cardiac death in Japan, especially among people in the prime of life, we investigated 133 out of 161 autopsied patients (82.6%) (106 men and 27 women, mean age 47.5 years). Coronary artery disease (CAD) was the most frequently detected disorder (50 cases, 37.6%), and included 15 cases of acute myocardial infarction (AMI) (11.3%). We found that CAD was less frequent among younger patients than in Western countries: 10.0% in subjects in their twenties and 22.2% among subjects in their thirties. The left anterior descending artery (LAD) was the vessel most often affected by infarction (47.0%), but the proportion of LAD lesions was not different from that in AMI patients who were survived for least 1 day after the attack. In conclusion, CAD was infrequent among patients aged 20-39 years in comparison with Western countries and LAD was the most commonly affected vessel, but the proportion was not different from that found among AMI patients who survived for at least 1 day after the attack.

Adult↗

Cladinose analogues of sixteen-membered macrolide antibiotics. IV. Improved therapeutic effects of 4-O-acyl-L-cladinose analogues of sixteen-membered macrolide antibiotics.

Six derivatives of sixteen-membered macrolides possessing 4-O-acyl-alpha-L-cladinose as a neutral sugar were synthesized via 3"-methylthiomethyl ether intermediates in reasonable yield. Introduction of a methyl group on the 3"-hydroxyl group of midecamycin A1 was effective for enhancing its antibacterial activity. All these derivatives exhibited excellent therapeutic effects in mice, and some of them showed improved pharmacokinetics compared with the natural antibiotics (mycarose type) in mice. Facile synthesis of 9-O-acylated analogues are also described.

Animals↗

Cladinose analogues of sixteen-membered macrolide antibiotics. III. Efficient synthesis of 4-O-alkyl-L-cladinose analogues: improved antibacterial activities compatible with pharmacokinetics.

The synthesis and biological evaluation of sixteen-membered macrolides possessing a 4-O-alkyl-alpha-L-cladinosyl moiety as a neutral sugar are described. These potent novel derivatives have been efficiently synthesized avoiding glycosylations. Two hydroxyl groups in mycarose of the tri-(tert-butyldimethylsilyl) ether intermediate were successively alkylated. Sequential deprotections of silyl groups afforded 4-O-alkyl-L-cladinose analogues and 3,4-di-O-alkyl-L-mycarose analogues of leucomycin V. Some 4-O-alkyl-L-cladinose analogues exhibited potent antibacterial activities. The most active derivative, 3"-O-methyl-4"-O-(3-methylbutyl)leucomycin V, showed improved metabolic stability in rat plasma in vitro and extremely high concentrations in serum after oral administrations in mice and in hamsters.

Animals↗