Biomedical subjects

K Sasamoto

Publications and source records attributed to K Sasamoto.

At least 19 recordsLinked to original sources

PubMed2003

Determination of stale-flavor carbonyl compounds in beer by stir bar sorptive extraction with in-situ derivatization and thermal desorption-gas chromatography-mass spectrometry.

A method for the determination of stale-flavor carbonyl compounds including E-2-octenal, E-2-nonenal, E,Z-2,6-nonadienal and E,E-2,4-decadienal in beer was developed using stir bar sorptive extraction (SBSE) with in-situ derivatization followed by thermal desorption-GC-MS analysis. The derivatization conditions with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine and the SBSE conditions--sampling mode, salt addition, sample volume, polydimethylsiloxane volume (sample/polydimethylsiloxane phase ratio) and extraction time--were examined. The method showed good linearity over the concentration range from 0.1 to 10 ng ml(-1) for all analytes and the correlation coefficients were higher than 0.9993. The limits of detection ranged from 0.021 to 0.032 ng ml(-1) for all analytes. The recoveries (98-101%) and precision (RSD 2.4-7.3%) of the method were examined by analyzing beer samples fortified at the 0.5-ng ml(-1) level. The method was successfully applied to low-level concentration samples.

Beer↗

PubMed2002

Neural activities in the substantia nigra modulated by stimulation of the orofacial motor cortex and rhythmical jaw movements in the rat.

Neurons related to jaw movements in the substantia nigra pars reticulata were explored by examining changes in their neural activities in response to electrical stimulation of the orofacial sensorimotor cortex and during rhythmical jaw movements induced by mechanical stimulation applied to the oral cavity in the rat. Out of 80 neurons tested, 59 showed changes in their firing patterns of activities in response to the electrical stimulation of the cortex. The responding neurons were mainly located in the dorsolateral part of the substantia nigra pars reticulata. The substantia nigra pars reticulata neurons showing responses were classified into the following five types according to their response patterns: (1) an inhibition preceded by an early excitation and followed by a late excitation (n = 26), (2) an inhibition preceded by an early excitation but not followed by a late excitation (n = 7), (3) an inhibition not preceded by an early excitation but followed by a late excitation (n = 2), (4) an inhibition without early or late excitations (n = 7) and (5) an excitation without an inhibition (n = 17). Out of 18 neurons responding to the cortical stimulation, 11 (61.1%) increased or decreased their neural activities during rhythmical jaw movements. Some of these neurons had a projection to the lateral part of the superior colliculus (n = 5) and/or to the parvicellular reticular formation (n = 2). These results provide first neurophysiological evidence for neurons in the dorsolateral part of the substantia nigra pars reticulata with inputs from and outputs to the areas related to jaw movements. These neurons may participate in the control of jaw movements in the rat.

Action Potentials↗

PubMed2001

Deficiency of triad junction and contraction in mutant skeletal muscle lacking junctophilin type 1.

In skeletal muscle excitation-contraction (E-C) coupling, the depolarization signal is converted from the intracellular Ca2+ store into Ca2+ release by functional coupling between the cell surface voltage sensor and the Ca2+ release channel on the sarcoplasmic reticulum (SR). The signal conversion occurs in the junctional membrane complex known as the triad junction, where the invaginated plasma membrane called the transverse-tubule (T-tubule) is pinched from both sides by SR membranes. Previous studies have suggested that junctophilins (JPs) contribute to the formation of the junctional membrane complexes by spanning the intracellular store membrane and interacting with the plasma membrane (PM) in excitable cells. Of the three JP subtypes, both type 1 (JP-1) and type 2 (JP-2) are abundantly expressed in skeletal muscle. To examine the physiological role of JP-1 in skeletal muscle, we generated mutant mice lacking JP-1. The JP-1 knockout mice showed no milk suckling and died shortly after birth. Ultrastructural analysis demonstrated that triad junctions were reduced in number, and that the SR was often structurally abnormal in the skeletal muscles of the mutant mice. The mutant muscle developed less contractile force (evoked by low-frequency electrical stimuli) and showed abnormal sensitivities to extracellular Ca2+. Our results indicate that JP-1 contributes to the construction of triad junctions and that it is essential for the efficiency of signal conversion during E-C coupling in skeletal muscle.

Animals↗

PubMed2001

Determination of trace amounts of off-flavor compounds in drinking water by stir bar sorptive extraction and thermal desorption GC-MS.

A method for the determination of trace amounts of off-flavor compounds including 2-methylisoborneol, geosmin and 2,4,6-trichloroanisole in drinking water was developed using the stir bar sorptive extraction technique followed by thermal desorption-GC-MS analysis. The extraction conditions such as extraction mode, salt addition, extraction temperature, sample volume and extraction time were examined. Water samples (20, 40 and 60 ml) were extracted for 60-240 min at room temperature (25 degrees C) using stir bars with a length of 10 mm and coated with a 500 microm layer of polydimethylsiloxane. The extract was analyzed by thermal desorption-GC-MS in the selected ion monitoring mode. The method showed good linearity over the concentration range from 0.1 or 0.2 or 0.5 to 100 ng l(-1) for all the target analytes, and the correlation coefficients were greater than 0.9987. The detection limits ranged from 0.022 to 0.16 ng l(-1). The recoveries (89-109%) and precision (RSD: 0.80-3.7%) of the method were examined by analyzing raw water and tap water samples fortified at the 1 ng l(-1) level. The method was successfully applied to low-level samples (raw water and tap water).

Gas Chromatography-Mass Spectrometry↗

PubMed2001

Analysis of rhythmical jaw movements produced by taste stimulation in rats.

Taste stimulation of the mouth induces various oral movements. Sucrose or salt solution induces rhythmical jaw movements (RJM) or tongue protrusion as an ingestive behavior. Bitter taste induces a gaping or tongue retraction as an aversive behavior. There is no report that describes the precise pattern of jaw movements induced by taste stimulation. The purpose of the present study is to clarify the pattern of the taste-induced RJM with electromyographic activities of the masticatory muscles in the anesthetized rat. When water was injected into the mouth of the rat, an RJM was induced. In this type of RJM, the lower jaw swung right and left side in each open-close cycle alternately. The anterior digastric muscle was active in every opening phase, while activities of the jaw closing muscles were generally weak. The RJM induced by water was opening-dominant movements. Sucrose or salt solution induced a similar pattern of RJM to water-induced RJM. When acetic acid was injected, amplitude of the alternate lateral jaw movement was significantly larger than that in the water-induced RJM. The activity of the superficial temporalis muscle was large in those closing phases with ipsilateral side movement, while it was small in the closing phases with contralateral movement. The pattern of quinine-induced RJM was characterized by small lateral, large open-close and large antero-posterior movements. Tastes which are reported to induce ingestive behavior such as sweet or salty don't alter the pattern of RJM; however, the tastes which induce aversive behavior make the pattern of RJM different from the water-induced RJM.

Animals↗

PubMed2001

Behavioral taste similarities and differences among monosodium L-glutamate and glutamate receptor agonists in C57BL mice.

Monosodium L-glutamate (MSG) and 5'-ribonucleotides elicit umami taste in humans and probably in some species of animals. Previous studies suggest that taste-mGluR4 and NMDA receptor may be involved in taste transduction for umami, but behavioral responses in rats do not support the involvement of NMDA receptor. In the present study, behavioral similarities and differences among MSG, mGluR4 agonist L(+)-2-amino-4-phosphonobutyrate (L-AP4), and NMDA receptor agonist N-methyl-D-aspartate (NMDA) were compared in C57BL mice by using a conditioned taste aversion paradigm. Mice conditioned to avoid either MSG or 10 mM L-AP4 appeared to avoid MSG, disodium 5'-inosinate (IMP), a mixture of MSG and IMP, and L-AP4, but not NMDA. Aversive conditioning to either sucrose or NMDA was generalized only to a mixture of MSG+IMP or NaCl. However, aversive conditioning to L-AP4 at 1 mM was generalized to NMDA and the umami substances. Lick rates for L-AP4 increased by mixing with (RS)-alpha-cycloprophy-4-phosphonophenylglycine (mGluR4 antagonist) when animals were conditioned to avoid MSG or L-AP4. Lick rates for NMDA also either decreased or increased by mixing with glycine (NMDA receptor coagonist) or D(-)-2-amino-5-phosphonopentanoic acid (NMDA receptor antagonist) when animals were conditioned to avoid L-AP4 or NMDA. In sucrose-conditioned mice. gurmarin (a sweet inhibiting peptide) suppressed the avoidance of sucrose and a mixture of MSG and IMP, but not L-AP4 and NMDA. The results suggest the possibility that to C 57BL mice MSG may taste similar to L-AP4 but different from NMDA, although both types of glutamate receptors as well as gurmarin-sensitive sweet receptor may be involved in perception of umami taste.

Animals↗

PubMed2001

Beta-cyclodextrin inhibits the sweet taste suppressing activity of gurmarin by the formation of an inclusion complex with aromatic residues in gurmarin.

Our recent study in mice revealed that the inhibitory activity of gurmarin on the sweet taste responses was reduced significantly by the presence of beta-cyclodextrin (beta-CD). To investigate the mechanism involved in the action of beta-CD, physicochemical experiments were performed on the interaction of CDs with gurmarin examining the effect of CDs on the UV absorption spectrum of gurmarin and on the elution behavior in gel filtration (or exclusion) chromatography. Among the three kinds of cyclodextrins tested, beta-CD induced significant changes in the UV absorption spectrum of gurmarin that were characteristic of those found in the inclusion complex formation of tyrosine and tryptophan with beta-CD. The abnormal retention behavior of gurmarin in gel filtration resulting from hydrophobic interaction with the gel matrix reverted to normal in the presence of beta-CD in the elution buffer. These results suggest that the unique domain of gurmarin, in which five aromatic amino acid residues are all directed outwardly and form a hydrophobic cluster, is a possible site of interaction with the gurmarin-sensitive sweet taste receptor molecules in rodents.

Chemical Phenomena↗

PubMed2000

[Histological identification of brainstem neurons related to generation of rhythmical jaw movements in the rat].

Rhythmical jaw movements (RJM) were induced by stimulation of the oral cavity or the cerebral peduncle in the anesthetized rat. We found that the phase-dependent rhythmical (RJM-phasic) neuronal activities were necessary for the generation of RJM and iontophoretic application of glutamate activated a small number of these neurons rhythmically in phase with simultaneous RJM (Glu-RJM-phasic neurons) although it activated most RJM-phasic neurons tonically without RJM. N-(2-aminoethyl) biotinamide hydrochloride was iontophoretically applied to visualize these neurons. An identified Glu-RJM-phasic neuron had the complicated outline of soma with eight or more short and long dendrites. The RJM-phasic neurons had the simple outlines of soma (fusiform, triangular or rhomboid) with several short dendrites. Differential activation by glutamate corresponded to differential morphological characteristics of the brainstem neurons related to RJM-generation.

Animals↗

PubMed1999

Activity of the rat pontomedullary reticular neurons related to rhythmical jaw movements.

Electrical stimulation of the cerebral peduncle or oral mechanical or chemical stimulation induced rhythmical jaw movements (RJM) in the anesthetized rat. Extracellular recording was made from the pontomedullary reticular neurons in relation to RJM. The RJM-related activity was classified to a tonic, a transient and a phase-dependent rhythmical activities (116, 92 and 45 neurons, respectively). We found that the neurons showing the phase-dependent activities discharged exclusively or almost exclusively during RJM without responding to at least one of three kinds of RJM-inducing stimulation. Fourteen neurons were activated by all kinds of RJM-inducing stimulation and majority of them showed weak or no change in activity during RJM, although some showed the transient or the rhythmical activity during RJM. We propose that the phase-dependent rhythmical activity is necessary to generate RJM since any stimulation-induced RJM was reversibly blocked by microinjection of lidocaine or glutamate receptor blocking agents into the brainstem site concentrated by this type of neurons. No other type of activity was observed exclusively during RJM.

Animals↗

PubMed1999

Glutamate activates neuronal discharges and rhythmical jaw movements in the rat.

Rhythmical jaw movements (RJM) were induced by stimulation of the cerebral peduncle or the oral cavity in the anesthetized rat and iontophoretic application of glutamate was examined on each reticular neuron with phase-dependent rhythmical activity during RJM (RJM-phasic neuron). Glutamate activated six neurons rhythmically with simultaneous rhythmical jaw movements in five rats. However, glutamate activated fifty of the other 52 RJM-phasic neurons tonically without RJM. Therefore, a small number of RJM-phasic neurons (10.3%) was involved in the generation of RJM. The rhythmical activities of most RJM-phasic neurons should have been synaptically induced by activities of those RJM-generator neurons. Glutamate receptor blocking agents, cyano-nitroquinoxaline-dione and D-amino-phosphono-valerate were examined and the former was more effective on 17 neurons although the latter was more effective on three and both were equally effective on 13. Inhibitory synaptic mechanism is not necessary for RJM-generation since bicuculline or strychnine did not significantly alter the rhythmical pattern of activity in any neuron.

Animals↗

PubMed1998

Neuronal activity related to spontaneous and capsaicin-induced rhythmical jaw movements in the rat.

Intraoral capsaicin induced rhythmical jaw movements (RJM) in anesthetized rats. Neurons in the trigeminal spinal nucleus caudalis or the cortico-peduncular (CP) axons were extracellularly recorded. Capsaicin excited dose-dependently most caudalis neurons, which were activated by stimulation of the oral cavity and/or the tooth pulp and activated during spontaneous or induced RJM. Ten of 55 CP axons were antidromically activated by stimulation of the contralateral trigeminal motor nucleus. All antidromic and 29 other CP axons discharged prior to the spontaneous RJM, but most of them did not during capsaicin-induced RJM. These neuronal activities possibly initiate spontaneous RJM although the activities of caudalis neurons are necessary for capsicin-induced RJM.

Animals↗

PubMed1998

[Effects of institutional therapy and factors which influence the prognosis].

We examined the effect of institutional therapy on asthmatic children and evaluating the factors influencing the prognosis. We analyzed the questionnaires and laboratory findings obtained form the 565 children who had been treated with the therapy in National Higashi-Saitama Hospital from April 1978 to March 1997. The institutional therapy seems to be effective in the following 3 aspects. 1) Pulmonary function test such as FEV1.0, and bronchial hypersensitivity test such as acetylcholine inhalation test were improved. 2) Exercise abilities such as free running were improved. 3) More than 40% of the patients had no asthma attack or only a few mild asthma attacks requiring no medication. When compared the group whose symptoms were not improved with the remission group, who had no asthma attack, significant differences were seen in the following points, 1) The dose and the number of the medicines at the end of the hospitalization, 2) the bronchial hypersensitivity test, 3) the age of onset, 4) the age of the symptoms become perennial and 5) the term from the symptoms become perennial to the hospitalization were significantly different. The therapy principles including the recipes for exercise training were not different between the two groups. It is concluded that we should consider institutional therapy or, a combination of physical training and environmental control as one of the most important therepeutic methods when asthmatic symptoms become perennial in spite of the conventional medication.

Asthma↗

PubMed1997

Evaluation of precipitation and direct methods for HDL-cholesterol assay by HPLC.

HDL-cholesterol (HDL-C) values measured by precipitation (sodium phosphotungstate-MgCl2) and direct methods were compared with those obtained by HPLC with a new column (TSKgel Lipopropak) and an eluent (TSKeluent LP-1). The HDL-C values determined by the precipitation method were significantly (P <0.001) lower than those by the HPLC method, whereas the HDL-C values by the direct method were slightly but significantly higher (P <0.02) than those by the HPLC method. A quantitative HPLC analysis of the cholesterol concentration in HDL and non-HDL fractions in the supernatant of serum separated by precipitation reagents with different MgCl2 concentrations ranging from 7.3 to 44 mmol/L revealed that reagents with >22 mmol/L MgCl2 precipitated part of HDL as well as non-HDL lipoproteins. The HPLC method providing quantitative and qualitative information with high precision was regarded as being a reliable approach for HDL-C assay. The HPLC can be also used to evaluate alternative methods for cholesterol assay.

Chemical Precipitation↗

PubMed1996

A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet.

Cell viability and in vitro cytotoxicity assay methods were developed using a combination of dyes, 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1), neutral red (NR) and crystal violet (CV), with HeLa cells as a bioindicatior. As WST-1 produces a highly water soluble and non-cytotoxic formazan dye, ti allows each assay to be carried out in one culture dish. The combined cell viability assay using WST-1, NR and CV gave an absorbance that correlated linearly with the number of cells over the range 1000 to 50,000 cells/well. The combined assay was applied to the evaluation of IC50 values for sodium dodecyl sulfate as a model toxicant, which yielded similar values to those obtained with each assay independently.

Cell Survival↗

PubMed1995

2-(5-hydrazinocarbonyl-2-oxazolyl)-5,6-dimethoxybenzothiazole as a precolumn fluorescence derivatization reagent for carboxylic acids in high-performance liquid chromatography and its application to the assay of fatty acids in human serum.

A unique acid hydrazide, 2-(5-hydrazinocarbonyl-2-oxazolyl)-5,6-dimethoxybenzothia zole, that is characterized by the benzothiazole structure conjugated to an oxazoline moiety was synthesized, and its applicability as a precolumn derivatization reagent for carboxylic acids in HPLC was examined in view of sensitivity and separability. The sensitivity of the hydrazide for carboxylic acids was determined using lauric acid to be 0.1 pmol (per 10-microliters injection volume) at a signal-to-noise ratio of 3. The reagent allowed rapid assays of carboxylic acids within 20 min with satisfactory separability. The method was applied to the determination of fatty acids in human sera from healthy volunteers as well as from patients with diabetes or thyroid dysfunction.

Carboxylic Acids↗

PubMed1995

[On the antibacterial effect of hinokitiol against Staphylococcus aureus showing double zone phenomenon].

We studied the antibacterial effect of Hinokitiol against Staphylococcus aureus. As reference, we studied also the antibacterial effects of the oil and water from Thujopsis dolablata var. hondai Makino which contains Hinokitiol an important element of its antibacterial effect. Interestingly, Hinokitiol showed the double zone phenomenon, i.e., minimum inhibitory concentration of hinokitiol against MSSA was 15-125 micrograms/ml and against MSSA was 125 micrograms/ml. But when the concentration of Hinokitiol was diluted to 1.87-0.94 micrograms/ml, then the growth of MSSA was inhibited (Table 1 and Fig. 1). This phenomenon was observed the study on antibacterial effect using disk containing Hinokitiol (Fig/3). Similar phenomenon were observed also in the study using water from Thujopsis dolablata (Table 2 & Fig. 2). We deduce that Hinokitiol has a strong antibacterial effect against S. aureus. But this effect shows double zone effect. The antibacterial effect of Hinokitiol is more prominent against MRSA than MSSA.

Dose-Response Relationship, Drug↗

PubMed1995

Development of a protein-free medium with iron salts replacing transferrin for a human-human hybridoma.

Many protein-free media have been developed, because protein-free media are usually more economical than serum-free or serum-containing media and facilitate the purification of bioactive materials. We evaluated various iron salts and chelating agents replacing transferrin to develop a protein-free medium for a human-human hybridoma, HB4C5, and found out that ferric citrate was favorable for the production and the productivity of monoclonal antibodies.

Antibodies, Monoclonal↗

PubMed1994

Dropout of young non-insulin-dependent diabetics from diabetic care.

We investigated the follow-up status and prognosis of 109 patients with non-insulin-dependent diabetes mellitus aged from 10 to 19 years old (66 males and 43 females). Patients who had not attended hospital for at least 20 months up to the end of September 1990 were regarded as defaulters, and were surveyed by questionnaire. There were 62 defaulters (56.9%) among the 109 patients originally enrolled in diabetes care. The defaulters had a significantly higher body mass index (both males and females), mean arterial blood pressure, and fasting blood glucose level than the patients still attending the diabetes clinic, as well as a significantly worse lipid profile. The main reason given for non-attendance was a busy schedule. Compared with patients attending the diabetes clinic, a lower percentage of the defaulters remained on a diet or took regular exercise. Rapid eating was more common among the defaulters than the attendees (92.9% vs. 60%, P = n.s.). Thus, the lifestyle of the defaulters seemed to be undesirable for young diabetic patients. These findings emphasize the importance of effective education and follow-up for young obese patients with non-insulin-dependent diabetes mellitus.

Adolescent↗