PubMed Health⌕ Search

Biomedical subjects

T Onaka

Publications and source records attributed to T Onaka.

At least 37 records · Page 2Linked to original sources
PubMed2001

Desulfurization characteristics of thermophilic Paenibacillus sp. strain A11-2 against asymmetrically alkylated dibenzothiophenes.

The thermophilic bacterium Paenibacillus sp. A11-2, which can utilize dibenzothiophene (DBT) as the sole sulfur source at high temperature (45-55 degrees C), was investigated for its ability to cleave carbon-sulfur bonds in the dibenzothiophene (DBT) ring with asymmetrical alkyl substitution, such as methyl, dimethyl, trimethyl, ethyl and propyl DBTs. The biodesulfurization products of each of these alkylated DBTs (Cx-DBTs) were identified and quantitatively determined. The results suggested that each of the Cx-DBTs was desulfurized at a low rate, then converted to alkylated hydroxybiphenyls containing the isomers, and molar ratios of these metabolic isomers were altered in terms of not only the positions but also the numbers and lengths of the alkyl substituents. Moreover, these ratios were compared with those obtained using the mesophilic desulfurizing bacterium Rhodococcus erythropolis KA2-5-1. Consequently, biodesulfurization reactions of these microbes could be characterized using asymmetrically Cx-DBTs and their molecular shape parameters (length and length-to-breadth ratio), indicating differences in the selectivity of the microbial enzymic systems between the two bacterial strains.

Journal Article↗
PubMed2001

Selective cleavage of the two CS bonds in asymmetrically alkylated dibenzothiophenes by Rhodococcus erythropolis KA2-5-1.

The Rhodococcus erythropolis strain KA2-5-1 was characterized by its ability to cleave carbon-sulfur bonds in the dibenzothiophene (DBT) ring by asymmetrically alkyl substitution, such as C2-DBTs (e.g., dimethyl and ethyl DBTs) and C3-DBTs (e.g., trimethyl and propyl DBTs), which are known to remain in hydrodesulfurization-treated diesel fuels. After treatment by solid-phase extraction (SPE) of solvents from microbial reactions of alkylated DBTs (Cx-DBTs), we used gas chromatography (GC), GC-atomic emission detection, GC-mass spectrometry and 1H nuclear magnetic resonance spectroscopy to identify and quantitatively evaluate the Cx-DBT metabolites. Molar ratios of metabolic isomers of the desulfurization products suggested that resting-cell reactions of KA2-5-1 against these Cx-DBTs occurrs through specific carbon-sulfur-bond-targeted cleavages, yielding alkylated hydroxybiphenyls, and that the manner of the attack on the DBT skeleton is affected not only by the position but also by the number and length of the alkyl substituents.

Journal Article↗
PubMed2000

Desulfurization of alkylated forms of both dibenzothiophene and benzothiophene by a single bacterial strain.

Thirty-five bacterial strains capable of converting dibenzothiophene into 2-hydroxybiphenyl were isolated. Among them Rhodococcus erythropolis KA2-5-1 was chosen for further characterization because of its ability to retain high desulfurization activity stably. PCR cloning and DNA sequencing of a KA2-5-1 genomic DNA fragment showed that it was practically identical with dszABC genes from Rhodococcus sp. IGTS8, a representative carbon-sulfur-bond-targeted dibenzothiophene-degrading bacterium. KA2-5-1 desulfurized a variety of alkyl dibenzothiophenes through the specific cleavage of their C-S bonds. In addition, unexpectedly, KA2-5-1 also attacked alkyl benzothiophenes in a C-S-bond-targeted fashion. The purified monooxygenase, encoded by dszC of KA2-5-1, converted benzothiophene and dibenzothiophene into benzothiophene sulfone and dibenzothiophene sulfone, respectively, with the aid of an NADH-dependent oxidoreductase. This result raises the possibility that the same enzymatic step may be involved in desulfurization of alkylated forms of both dibenzothiophene and benzothiophene in KA2-5-1 cells.

Alkylation↗
PubMed2000

Demonstration of the carbon-sulfur bond targeted desulfurization of benzothiophene by thermophilic Paenibacillus sp. strain A11-2 capable of desulfurizing dibenzothiophene.

Paenibacillus sp. strain A11-2, which had been primarily isolated as a bacterial strain capable of desulfurizing dibenzothiophene to produce 2-hydroxybiphenyl at high temperatures, was found to desulfurize benzothiophene more efficiently than dibenzothiophene. The desulfurized product was identified as o-hydroxystyrene by GC-MS and 1H-NMR analysis. Benzothiophene was assumed to be degraded in a way analogous to the 4S pathway, which has been well-known as a mode of dibenzothiophene degradation. These results suggest that benzothiophene desulfurization may share at least partially the reaction mechanism with dibenzothiophene desulfurization.

Bacillus↗
PubMed2000

A Broad 22 Micron Emission Feature in the Carina Nebula H ii Region.

We report the detection of a broad 22 µm emission feature in the Carina Nebula H ii region by the Infrared Space Observatory (ISO) short-wavelength spectrometer. The feature shape is similar to that of the 22 µm emission feature of newly synthesized dust observed in the Cassiopeia A supernova remnant. This finding suggests that both of the features are arising from the same carrier and that supernovae are probably the dominant production sources of this new interstellar grain. A similar broad emission dust feature is also found in the spectra of two starburst galaxies from the ISO archival data. This new dust grain could be an abundant component of interstellar grains and can be used to trace the supernova rate or star formation rate in external galaxies. The existence of the broad 22 µm emission feature complicates the dust model for starburst galaxies and must be taken into account correctly in the derivation of dust color temperature. Mg protosilicate has been suggested as the carrier of the 22 µm emission dust feature observed in Cassiopeia A. The present results provide useful information in studies on the chemical composition and emission mechanism of the carrier.

Journal Article↗
PubMed2000

Operon structure and functional analysis of the genes encoding thermophilic desulfurizing enzymes of Paenibacillus sp. A11-2.

Paenibacillus A11-2 can efficiently cleave two carbon&bond;sulfur bonds in dibenzothiophene (DBT) and alkyl DBTs, which are refractory by conventional petroleum hydrodesulfurization, to remove sulfur atom at high temperatures. An 8.7-kb DNA fragment containing the genes for the DBT desulfurizing enzymes of A11-2 was cloned in Escherichia coli and characterized. Heterologous expression analysis of the deletion mutants identified three open reading frames that were required for the desulfurization of DBT to 2-hydroxybiphenyl (2-HBP). The three genes were designated tdsA, tdsB, and tdsC (for thermophilic desulfurization). Both the nucleotide sequences and the deduced amino acid sequences show significant homology to dszABC genes of Rhodococcus sp. IGTS8, but there are several local differences between them. Subclone analysis revealed that the product of tdsC oxidizes DBT to DBT-5,5'-dioxide via DBT-5-oxide, the product of tdsA converts DBT-5,5'-dioxide to 2-(2-hydroxyphenyl) benzene sulfinate, and the product of tdsB converts 2-(2-hydroxyphenyl)benzene sulfinate to 2-HBP. Cell-free extracts of a recombinant E. coli harboring all the three desulfurization genes converted DBT to 2-HBP at both 37 and 50 degrees C. In vivo and in vitro exhibition of desulfurization activity of the recombinant genes derived from a Paenibacillus indicates that an E. coli oxidoreductase can be functionally coupled with the monooxygenases of a gram-positive thermophile.

Amino Acid Sequence↗
PubMed2000

Application of solid-phase extraction to the analysis of the isomers generated in biodesulfurization against methylated dibenzothiophenes.

A solid-phase extraction (SPE) technique was applied to analyze and characterize the biodesulfurization reactions against asymmetrically methylated dibenzothiophenes (mDBTs) such as 1-, 2-, 3- and 4-methyldibenzothiophenes present in fossil fuels. Recently, we found that these mDBTs are efficiently degraded by the bacterial strain, Rhodococcus erythropolis KA2-5-1. Separation and concentration of the microbial desulfurization products from each of the mDBTs could be carried out with high efficiency and reproducibility by the SPE procedure. These desulfurization products were identified using the SPE technique combined with GC, GC-atomic emission detection, GC-MS and 1H nuclear magnetic resonance spectroscopy. The analytical data obtained suggested that the desulfurization reactions against mDBTs by this bacterial strain may occur through specific carbon-sulfur bond-targeted cleavages that can be affected by the positions of methyl groups.

Chromatography, Gas↗
PubMed2000

Vasopressin regulation of noradrenaline release within the supraoptic nucleus.

The effect of electrically evoked dendritic vasopressin release on noradrenaline release into the hypothalamic supraoptic nucleus was assessed by in vivo microdialysis in conjunction with high pressure liquid chromatography and electrochemical detection. Electrical activation of magnocellular supraoptic neurones by stimulation of their axons at the level of the neural lobe significantly increased noradrenaline release into the nucleus (2.5-fold, P<0.03). This increase was completely blocked by administration of a nonpeptide vasopressin V1a receptor antagonist via the microdialysis probe. These data suggest that dendritically released vasopressin facilitates noradrenaline release into the hypothalamic nucleus.

Antidiuretic Hormone Receptor Antagonists↗
PubMed2000

Catecholaminergic mechanisms underlying neurohypophysial hormone responses to unconditioned or conditioned aversive stimuli in rats.

Oxytocin release from the neurohypophysis is facilitated by systemic cholecystokinin octapeptide (CCK) administration and noxious stimuli. Oxytocin release after CCK administration is mediated by A2 noradrenergic neurones while the release after noxious stimuli appears to be mediated by A1 noradrenergic neurones. On the other hand, facilitation of vasopressin release after noxious stimuli is not dependent upon noradrenergic neurones but on dopamine receptors. Environmental stimuli previously paired with noxious stimuli (conditioned fear stimuli) or novel environmental stimuli facilitate oxytocin release and suppress vasopressin release. These neuroendocrine responses to conditioned fear stimuli, but not to novel stimuli, are impaired by central noradrenaline depletion or i.c.v. adrenoceptor antagonists. These data suggest that there are at least two types of stress responses in neuroendocrine systems, one noradrenaline dependent, and one noradrenaline independent. It is also suggested that noradrenergic neurones are functionally heterogeneous in the control of oxytocin release.

Animals↗
PubMed2000

Purification and characterization of dibenzothiophene sulfone monooxygenase and FMN-dependent NADH oxidoreductase from the thermophilic bacterium Paenibacillus sp. strain A11-2.

A dibenzothiophene (DBT) sulfone monooxygenase (TdsA), which catalyses the oxidative CS bond cleavage of DBT sulfone to produce 2-(2-hydroxyphenyl)benzenesulfinate (HPBS) was purified from the thermophilic DBT desulfurizing bacterium Paenibacillus sp. strain A11-2 by multistep chromatography. The molecular mass of the purified enzyme was determined to be 120 kDa by gel filtration and the subunit molecular mass was calculated to be 48 kDa by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) indicating a dimeric structure. The N-terminal amino acid sequence of the purified TdsA was determined to be MRQMHLAGFFAAGNTHH, which revealed no significant similarity to any other known amino acid sequences. The purified TdsA absolutely required an oxidoreductase for its activity. This oxidoreductase (TdsD) was also purified to homogeneity, and its molecular size was calculated to be 50 kDa and 25 kDa by gel filtration and SDS-PAGE, respectively. TdsD was completely FMN-dependent, and FAD could not act as a cofactor. The N-terminal amino acid sequence of the purified TdsD was determined to be TSQTAEQSIAPIVAQYRHPEQPISALFVNR, which showed significant similarity to kinesin-like protein (44% identity). The optimal temperatures for the activity of TdsA and TdsD were 45 degrees C and 55 degrees C, respectively. Both enzymes showed optimal activity at pH 5.5. TdsA was slightly inhibited by sulfate, but not by 2-hydroxybiphenyl (2-HBP), which is another end product of DBT. TdsA showed higher activity toward bulkier substrates than its mesophilic counterpart, DszA. These properties suggest the applicability of biodesulfurization to the processing of actual petroleum fractions.

Journal Article↗
PubMed2000

Alkylated benzothiophene desulfurization by Rhodococcus sp. strain T09.

A benzothiophene desulfurizing bacterium was isolated and identified as Rhodococcus sp. strain T09. Growth assays revealed that this strain assimilated, as the sole sulfur source, various organosulfur compounds that cannot be assimilated by the well-studied dibenzothiophene-desulfurizing Rhodococcus sp. IGTS8. The cellular growth rate of strain T09 for the alkylated benzothiophenes depended on the alkylated position and the length of the alkyl moiety.

Alkylation↗
PubMed1999

Does the pineal gland play a role in neuroendocrine fear responses?

The pineal gland secretes melatonin under an influence of suprachiasmatic nucleus neurones. Pinealectomy or melatonin administration affects behavioural responses to novel stimuli. Fear or novel stimuli inhibit vasopressin (VP) and facilitate oxytocin (OT) or prolactin (PRL) release from the pituitary. Thus the suprachiasmatic nucleus-pineal gland system may modulate VP, OT and PRL responses to conditioned fear stimuli. In the present experiments with male rats, pinealectomy or melatonin administration did not significantly change VP, OT or PRL responses to conditioned fear stimuli. Electrolytic lesions of the suprachiasmatic nuclei impaired VP but not OT or PRL responses. The results show that the pineal gland is not involved in neuroendocrine responses to conditioned fear stimuli and suggest that the suprachiasmatic nucleus is necessary for the VP response to fear stimuli.

Animals↗
PubMed1998

Role of noradrenergic projections to the bed nucleus of the stria terminalis in neuroendocrine and behavioral responses to fear-related stimuli in rats.

The bed nucleus of the stria terminalis (BNST) receives dense noradrenergic projections from the brainstem and has been claimed to play a role in expression of a variety of stress responses. Fear-related stimuli suppress vasopressin and facilitate oxytocin release from the neurohypophysis and induce behavioral suppression. Here we investigated in male rats whether conditioned fear stimuli increase noradrenergic activity in the BNST and whether depletion of epinephrine content in the BNST prevents neuroendocrine and behavioral responses to fear stimuli. Environmental stimuli previously paired with electric footshocks increased the ratio of 3-methoxy-4-hydroxyphenylglycol to norepinephrine contents in the BNST, suggesting that the stimuli activated noradrenergic projections to the BNST. 5-Amino-2, 4-dihydroxy-alpha-methylphenylethylamine, a neurotoxin relatively selective for noradrenergic fibers, when injected into the BNST 7 days before measurement, decreased the content of norepinephrine by 95% and that of dopamine or serotonin by about 50%. In the rats that received the neurotoxin, the suppressive vasopressin but not the augmentative oxytocin response to intermittent footshocks was abolished. In the experiments with conditioned fear stimuli, the neurotoxin given before training partially but significantly impaired the suppressive vasopressin and behavioral responses to testing stimuli. The neurotoxin given after training, however, did not prevent the vasopressin, oxytocin or behavioral responses. The results suggest that noradrenergic fibers in the BNST mediate the suppressive vasopressin but not the augmentative oxytocin response to nonassociatively applied fear stimuli and that they modulate, in a facilitative fashion, acquisition but not retention or recall of the emotional memory associated with the vasopressin and behavioral responses to conditioned fear stimuli.

Amygdala↗
PubMed1998

Role of NMDA receptors in the emotional memory associated with neuroendocrine responses to conditioned fear stimuli in the rat.

Behavioral experiments have shown that the N-methyl D-aspartate (NMDA) subclass of glutamate receptor plays an important role in acquisition of emotional memory. Exposure of a rat to conditioned fear stimuli suppresses vasopressin (VP) release and augments oxytocin (OT) or prolactin (PRL) release from the pituitary. Present experiments aimed at investigating the effect of intraperitonially administered MK-801, an antagonist of NMDA receptor on the emotional memory associated with the suppressive VP and the augmentative OT or PRL responses to conditioned fear stimuli in male rats. MK-801 injected 30 min before training impaired the VP, OT and PRL responses to the testing fear stimuli. The antagonist injected after training, however, did not block the responses. MK-801 administered before testing impaired the previously acquired VP, OT and PRL responses to conditioned fear stimuli. In the experiments with non-associatively applied fear stimuli, MK-801 did not block the VP, OT or PRL response. In the experiments with novel environmental stimuli, MK-801 did not impair VP, OT or PRL responses. The results suggest that an activation of NMDA receptors are required to acquire and recall but not to consolidate or retain the emotional memory associated with VP, OT and PRL responses to conditioned fear stimuli.

Animals↗
PubMed1998

Oxytocin release from the neurohypophysis after the taste stimuli previously paired with intravenous cholecystokinin in anaesthetized rats.

Intravenously administered cholecystokinin octapeptide (CCK) induces oxytocin release from the neurohypophysis in anaesthetised rats. Memory of conditioned taste aversion can be acquired under anaesthesia. The present experiments aimed at investigating whether taste stimuli previously paired with i.v. CCK evoke oxytocin release from the neurohypophysis in urethane-anaesthetised male rats. Sucrose solution (0.75-2.0 M) paired with i.v. CCK or the vehicle was applied to the tongue. After 3 h, sucrose solution was applied again. The second sucrose slightly increased plasma oxytocin concentration in rats that had received the first sucrose solution paired with the vehicle. Plasma oxytocin concentration after the second sucrose application, however, was significantly higher in CCK-injected than in vehicle-injected rats. In rats that received CCK 1 h before the first sucrose application, a second sucrose application did not produce the oxytocin response. The magnitude of the oxytocin response to the second sucrose solution was increased in a manner related to CCK doses. In separate experiments, NaCl solution (0.75 M) paired with CCK or the vehicle was applied to the tongue. The second NaCl solution applied 3 h after the first one facilitated oxytocin release both in the rats that had received CCK or the vehicle. The increase in plasma oxytocin, however, was significantly larger in CCK than in vehicle-injected rats. In rats that had received the first sucrose solution paired with CCK, a second sucrose solution evoked a significantly larger increase in plasma oxytocin concentrations than a testing NaCl solution did. In rats that had received NaCl solution paired with CCK, a testing sucrose solution did not significantly change plasma oxytocin concentrations. These data suggest that the taste stimulus previously paired with i.v. CCK induces oxytocin release from the neurohypophysis in urethane-anaesthetised rats.

Anesthetics, Intravenous↗
PubMed1998

Role of adrenoceptors in vasopressin, oxytocin and prolactin responses to conditioned fear stimuli in the rat.

Conditioned fear or novel environmental stimuli suppress vasopressin (VP) and augment oxytocin (OT) and prolactin (PRL) release in rats. We examined the effects of intracerebroventricular (i.c.v.) injections of adrenoceptor antagonists on these neuroendocrine responses to conditioned fear or novel environmental stimuli in male rats. A beta1 antagonist, metoprolol, blocked the VP but not the OT or PRL response to conditioned fear stimuli, but did not abolish neuroendocrine responses to novel environmental stimuli. A beta2 antagonist, ICI118551, impaired the PRL but not the VP or OT response to fear or novel environmental stimuli. In rats injected with a alpha1 adrenoceptor antagonist, benoxathian, conditioned fear stimuli did not significantly induce the VP, OT or PRL responses. The effects of benoxathian were not due to a general reduction of arousal, since benoxathian did not prevent the VP, OT or PRL response to novel environmental stimuli. These data suggest that beta1 adrenoceptors play a selective role in the VP response to conditioned fear stimuli, as do beta2 adrenoceptors in the prolactin response to conditioned fear and novel environmental stimuli. We conclude that alpha1 adrenoceptors play a facilitative role in VP, OT, PRL responses to conditioned fear stimuli.

Adrenergic Antagonists↗
PubMed1997

The role of afferent inputs to supraoptic nucleus oxytocin neurons during naloxone-precipitated morphine withdrawal in the rat.

During prolonged exposure to morphine, oxytocin neurons of the rat supraoptic nucleus develop dependence, shown by hyperexcitation following morphine withdrawal. The present study investigated the role of afferent projections to the supraoptic nucleus in this withdrawal excitation. Rats were made morphine-dependent by continuous intracerebroventricular infusion of morphine at increasing doses (up to 50 microg/h). On the sixth day, rats were anaesthetized with pentobarbitone and morphine withdrawal was precipitated by intraperitoneal injection of naloxone (5 mg/kg). Fos-immunoreactivity in the supraoptic nucleus, and also in the median preoptic nucleus, organum vasculosum of the lamina terminalis and subfornical organ, which project to the supraoptic nucleus, increased following morphine withdrawal. However, retrograde tracing from the supraoptic nucleus showed that, of the neurons in these regions which project to the supraoptic nucleus, only 0.4-7.1% expressed Fos in response to morphine withdrawal. Following morphine withdrawal, Fos-immunoreactivity was present in 39.2% and 19.8% of the tyrosine hydroxylase-immunoreactive neurons of the A1/C1 and A2/C2 cell groups. Of the cells in these regions identified as projecting to the supraoptic nucleus, 11.3% in the region of the A2 cell group and 12.7% in the region of the A1 cell group expressed Fos after morphine withdrawal. In a second study, monoamine release was measured in the supraoptic nucleus of urethane-anaesthetized morphine-dependent and -naive rats. Retrodialysis of naloxone (10[-5] M) into the supraoptic nucleus induced a small increase in plasma oxytocin concentration in morphine-dependent rats (13.5+/-4.8 pg/ml increase) but not in naive rats (1.2+/-5.9 pg/ml decrease), with no significant change in monoamine release in either morphine-dependent or -naive rats. Intravenous injection of naloxone (5 mg/kg) 1 h later produced a further significant increase in plasma oxytocin concentration in morphine-dependent rats concomitant with a significant increase in noradrenaline release from the supraoptic nucleus. Thus, morphine-withdrawal excitation of supraoptic oxytocin neurons occurs concurrently with a modestly increased activity of their input from the brainstem, and very little activation in other known inputs.

Afferent Pathways↗