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Biomedical subjects

A Nishi

Publications and source records attributed to A Nishi.

At least 37 records · Page 2Linked to original sources

DARPP-32: regulator of the efficacy of dopaminergic neurotransmission.

Dopaminergic neurons exert a major modulatory effect on the forebrain. Dopamine and adenosine 3',5'-monophosphate-regulated phosphoprotein (32 kilodaltons) (DARPP-32), which is enriched in all neurons that receive a dopaminergic input, is converted in response to dopamine into a potent protein phosphatase inhibitor. Mice generated to contain a targeted disruption of the DARPP-32 gene showed profound deficits in their molecular, electrophysiological, and behavioral responses to dopamine, drugs of abuse, and antipsychotic medication. The results show that DARPP-32 plays a central role in regulating the efficacy of dopaminergic neurotransmission.

Amphetamines↗

Bidirectional regulation of DARPP-32 phosphorylation by dopamine.

Dopamine has been shown to stimulate phosphorylation of DARPP-32, a phosphoprotein highly enriched in medium-sized spiny neurons of the neostriatum. Here, we investigated the contribution of D1-like and D2-like dopamine receptors in the regulation of DARPP-32 phosphorylation in mouse striatal slices. D1-like and D2-like receptors had opposing effects on the state of DARPP-32 phosphorylation. The D1 receptor agonist SKF82526 increased DARPP-32 phosphorylation. In contrast, the D2 receptor agonist quinpirole decreased basal as well as D1 agonist-, forskolin-, and 8-bromo-cAMP-stimulated phosphorylation of DARPP-32. The ability of quinpirole to decrease D1-stimulated DARPP-32 phosphorylation was calcium-dependent and was blocked by the calcineurin inhibitor cyclosporin A, suggesting that the D2 effect involved an increase in intracellular calcium and activation of calcineurin. In support of this interpretation, Ca2+-free/EGTA medium induced a greater than 60-fold increase in DARPP-32 phosphorylation and abolished the ability of quinpirole to dephosphorylate DARPP-32. The antipsychotic drug raclopride, a selective D2 receptor antagonist, increased phosphorylation of DARPP-32 under basal conditions and in D2 agonist-treated slices. The results of this study demonstrate that dopamine exerts a bidirectional control on the state of phosphorylation of DARPP-32.

8-Bromo Cyclic Adenosine Monophosphate↗

Functional analyses of a variety of chimeric dioxygenases constructed from two biphenyl dioxygenases that are similar structurally but different functionally.

The biphenyl dioxygenases (BP Dox) of strains Pseudomonas pseudoalcaligenes KF707 and Pseudomonas cepacia LB400 exhibit a distinct difference in substrate ranges of polychlorinated biphenyls (PCB) despite nearly identical amino acid sequences. The range of congeners oxidized by LB400 BP Dox is much wider than that oxidized by KF707 BP Dox. The PCB degradation abilities of these BP Dox were highly dependent on the recognition of the chlorinated rings and the sites of oxygen activation. The KF707 BP Dox recognized primarily the 4'-chlorinated ring (97%) of 2,5,4'-trichlorobiphenyl and introduced molecular oxygen at the 2',3' position. The LB400 BP Dox recognized primarily the 2,5-dichlorinated ring (95%) of the same compound and introduced O2 at the 3,4 position. It was confirmed that the BphA1 subunit (iron-sulfur protein of terminal dioxygenase encoded by bphA1) plays a crucial role in determining the substrate selectivity. We constructed a variety of chimeric bphA1 genes by exchanging four common restriction fragments between the KF707 bphA1 and the LB400 bphA1. Observation of Escherichia coli cells expressing various chimeric BP Dox revealed that a relatively small number of amino acids in the carboxy-terminal half (among 20 different amino acids in total) are involved in the recognition of the chlorinated ring and the sites of dioxygenation and thereby are responsible for the degradation of PCB. The site-directed mutagenesis of Thr-376 (KF707) to Asn-376 (LB400) in KF707 BP Dox resulted in the expansion of the range of biodegradable PCB congeners.

Amino Acid Sequence↗

Clinical quiz.

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Acute Disease↗

Recurrent exercise-induced acute renal failure in renal hypouricemia.

We describe a male patient with four episodes of acute renal failure after strenuous exercise occurring between the age of 14 and 25 years. He was found to have low serum uric acid (0.4 mg dl-1 after recovery) and high fractional excretion of uric acid. A benzbromarone/pyrazinamide test suggested that renal hypouricemia was due to defective proximal tubular reabsorption of uric acid at a presecretory site. A renal biopsy revealed acute tubular necrosis, a renal computer tomography scan showed patchy contrast enhancement and a treadmill exercise test induced an immediate fall in creatinine clearance. These findings suggest that the cause of acute renal failure was renal vasoconstriction rather than obstruction by uric acid crystals.

Acute Kidney Injury↗

Engineering hybrid pseudomonads capable of utilizing a wide range of aromatic hydrocarbons and of efficient degradation of trichloroethylene.

We constructed hybrid Pseudomonas strains in which the bphA1 gene (coding for a large subunit of biphenyl dioxygenase) is replaced with the todC1 gene (coding for a large subunit of toluene dioxygenase of Pseudomonas putida Fl) within chromosomal biphenyl-catabolic bph gene clusters. Such hybrid strains gained the novel capability to grow on a wide range of aromatic hydrocarbons, and, more interestingly, they degraded chloroethenes such as trichloroethylene and cis-1,2-dichloroethylene very efficiently.

Benzene Derivatives↗

Studies on metabolites of mycoparasitic fungi. V. Ion-spray ionization mass spectrometric analysis of trichokonin-II, a peptaibol mixture obtained from the culture broth of Trichoderma koningii.

The sequence of a peptide, trichokonin-II (TK-II), obtained from the culture broth of Trichoderma koningii OUDEMANS, was examined by ion-spray ionization mass spectrometry (ISI-MS), including the collision-induced dissociation (CID) technique. TK-II was concluded to be a mixture of three peptaibols, TK-IIa, TK-IIb, and TK-IIc.

Alamethicin↗

Longitudinal changes in peritoneal equilibration test with or without peritonitis in children.

This study was conducted to evaluate longitudinal changes in the peritoneal equilibration test (PET) in children treated with continuous peritoneal dialysis (CPD). The effects of prolonged CPD and episodes of peritonitis on the PET were examined. PET was repeated up to five times in 12 paediatric patients who were subdivided into groups with and without peritonitis. In the peritonitis group (n = 6), the dialysate/plasma (D/P) creatinine ratio at a 4-h dwell time decreased progressively with time on CPD in five of six patients. In a comparison of the initial and final PETs performed at a mean interval of 22.8 +/- 11.6 months, the D/P creatinine ratio in the final PET was significantly lower than in the initial PET (P < 0.01). In contrast, in the non-peritonitis group (n = 6), the D/P creatinine ratio in the final PET was unchanged for 28.2 +/- 12.3 months from the initial PET. The D/Do glucose ratio at a 4-h dwell time was unchanged over time in each group. Thus, repeated PET measurements revealed that membrane permeability for creatinine was not affected by prolonged CPD itself, but decreased with time after episodes of peritonitis. Although the protocol for PET is not standardised in children, PET was useful for determining the sequential changes in peritoneal function in such patients on CPD.

Child↗

Separation of bacterial cells by free flow electrophoresis under microgravity: a result of the SpaceLab-Japan project on Space Shuttle flight STS-47.

We demonstrated free flow electrophoresis (FFE) of charged cells under microgravity, where gravitational effects are almost eliminated. Separation of a mixture of three bacterial strains (mutants of Salmonella typhimurium LT2) by FFE was conducted on NASA Space Shuttle flight STS-47 (September 1992). The experiment was designed to differentiate three strains having different lipopolysaccharide core structures in the cell membrane. The results were compared to those of ground experiments, in order to examine whether or not FFE in a weightless environment provides distinct advantages. Smooth strain SL1027 and rough strain SL3749 migrated to two separated fractions. The quality (viability) and the yields of the separated samples were sufficient to show the advantage of microgravity. Another rough strain, SL1102, exhibited unexpected electrophoretic behavior, which prevented the complete resolution of the three strains. All the strains were recovered as viable cells after 8 days of flight. The present study suggests that electrophoretic separation of bacterial cells is much more effective under microgravity conditions with relatively good resolution in comparison with the ground operation.

Anti-Bacterial Agents↗

Physarum polycephalum haemagglutinins: effect of nutrition on synthesis, and their possible role in nature.

The activity of haemagglutinins in plasmodia of Physarum polycephalum was measured under different culture conditions. The activity was markedly increased when the plasmodia were incubated in a non-nutrient salt medium. During starvation, significant amounts of haemagglutinins were found in the slime layer on the surface of the plasmodia. An increase in activity was not observed in the presence of actinomycin D or cycloheximide. Under starvation conditions, plasmodia are known to differentiate into either sclerotia (spherules) or fruiting bodies. Acceleration of haemagglutinin synthesis, however, was not always observed during spherulation and fruiting-body formation. Attempts to detect endogenous glycoconjugates that bind to the haemagglutinins were unsuccessful but we found that the haemagglutinins could bind to acidic polysaccharides produced by Escherichia coli K12. The bacterial glycoconjugates were purified and partially characterized. They contained N-acetylhexosamine residues which appeared to be important for binding with the haemagglutinins. It is possible that the haemagglutinins play a physiological role in the interaction with these organisms.

Animals↗

Anti-neutrophil cytoplasmic antibody for proteinase 3 in a child with polyarteritis nodosa.

We report a case of a 15 year old boy with polyarteritis nodosa associated with antineutrophil cytoplasmic antibody (ANCA) against proteinase 3 (PR3). After months of steroid and immunosuppressant therapy, the symptoms subsided and the polyaneurysms almost disappeared. The levels of anti-PR3 antibody and of cytokines also decreased. The results indicate that ANCA is a good indicator of this disease activity, and may play some pathogenic role in the disease.

Adolescent↗

Studies on metabolites of mycoparasitic fungi. IV. Minor peptaibols of Trichoderma koningii.

Three minor peptaibols, trichokonins (TKs)-Ia, Ib, and IX, were obtained from the culture broth of Trichoderma koningii Oudemans. Primary structures of these peptaibols were elucidated by ion-spray ionization mass spectrometry (ISI-MS) including the collision-induced dissociation (CID) technique together with two-dimensional nuclear Overhauser enhancement spectroscopy (NOESY).

Alamethicin↗

Studies on metabolites of mycoparasitic fungi. II. Metabolites of Trichoderma koningii.

Four peptaibols, named trichokonins (TKs) V, VI, VII, and VIII, were isolated from the culture broth of Trichoderma koningii Oudemans. Primary structures of these peptaibols were elucidated by electrospray ionization mass spectrometry (ESI-MS), FAB-MS, and collision-induced dissociation (CID) techniques along with nuclear Overhauser enhancement spectroscopy (NOESY).

Alamethicin↗

Involvement of plasma membrane-located calmodulin in the response decay of cyclic nucleotide-gated cation channel of cultured carrot cells.

Increase in cytoplasmic cyclic AMP concentration stimulates Ca2+ influx through the cyclic AMP-gated cation channel in the plasma membrane of cultured carrot cells. However, the Ca2+ current terminated after a few minutes even in the presence of high concentrations of cyclic AMP indicating that hydrolysis of the nucleotide is not responsible for stop of the Ca2+ influx. Cyclic AMP evoked discharge of Ca2+ from inside-out sealed vesicles of carrot plasma membrane, and it was strongly inhibited when the suspension of the vesicles was supplemented with 1 microM of free Ca2+, while Ca2+ lower than 0.1 microM did not affect the Ca(2+)-release. The Ca2+ flux across plasma membrane was restored from this Ca(2+)-induced inhibition by the addition of calmodulin inhibitors or anti-calmodulin. These results suggest that Ca2+ influx initiated by the increase in intracellular cAMP in cultured carrot cells is terminated when the cytosolic Ca2+ concentration reaches the excitatory level in the cells, and calmodulin located in the plasma membrane plays an important role in the response decay of the cyclic nucleotide-gated Ca2+ channel.

Biological Transport↗