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Biomedical subjects

X Xin

Publications and source records attributed to X Xin.

64 records · Page 4Linked to original sources
PubMed1994

Probing the Vibrio harveyi luciferase beta subunit functionality and the intersubunit domain by site-directed mutagenesis.

While the critical role of the bacterial luciferase alpha subunit in catalysis has been amply documented, the beta subunit was only known to be involved in thermal stability and substrate binding. Two conserved histidyl residues at position 81 and 82 of the beta subunit of Vibrio harveyi luciferase were each mutated to an alanine, aspartate, or lysine to probe further the beta functionality. These mutations resulted in higher Km values for reduced riboflavin 5'-phosphate, less efficient oxidations of the aldehyde substrate, and decreased light-emitting activities. beta His82 appears to be significantly more critical than beta His81. For the beta His82-mutated luciferases, the maximal light intensities and total light outputs were reduced to 19-4% of that for the wild-type enzyme, and the values of Vmax/Km,flavin were decreased by 2-3 orders of magnitude. The reduced light emission activities for these mutated luciferases can be correlated to lower yields of the flavin 4a-hydroperoxide intermediate, reduced productions of the excited flavin emitter, and/or enhanced quenching of the emitter. The beta subunit and the conserved beta His82 in particular have thus been shown to be critical not only to flavin binding but also to catalytic characteristics of luciferase. The dimeric structure of luciferase is essential to its high catalytic efficiency. To characterize the intersubunit domain, three sets of single/double mutants were constructed, and the additivities of mutational effects were tested to screen for residues that could interact across the subunit interface.(ABSTRACT TRUNCATED AT 250 WORDS)

Crystallography, X-Ray↗
PubMed1994

Morphological changes in rat aortic endothelial cell line stimulated by endothelin.

We have reported that endothelin (ET)-1 activates interleukin (IL)-6 production in rat aortic endothelial cell line (WAE-1 cell). In this experiment, we investigated the morphological changes induced by ET-1 in cultured WAE-1 cells. The cells were treated with ET-1 for 8 h or 24 h, and then compared with untreated cells by light and electron microscopies. The WAE-1 cells treated with ET-1 for 8 h showed remarkable alterations as following: by light microscopic observation, the cells were enlarged and filled with many vesicles in the cytoplasm, and by electron microscopic observation, the cells showed the increases of nuclear membrane infoldings, increased density of chromatin granules just inside the nuclear membrane, and multivesicular bodies in the cytoplasm. These morphological changes were hardly observed in WAE-1 cells-treated with ET-1 for 24 h. It was suggested that ET-1 stimulated DNA synthesis and secretion of cytoplasmic products in WAE-1 cells.

Animals↗
PubMed1991

Functional consequences of site-directed mutation of conserved histidyl residues of the bacterial luciferase alpha subunit.

The available sequences for the different bacterial luciferases reveal five conserved histidyl residues at positions 44, 45, 82, 224, and 285 of the alpha subunit. Ten variants of Vibrio harveyi luciferase were obtained by selective site-directed mutations of these five histidines. The essentiality of alpha His44 and alpha His45 was indicated by 4-7 orders of magnitude of bioluminescence activity reductions resulting from the substitution of either histidine by alanine (alpha H44A or alpha H45A), aspartate (alpha H44D or alpha H45D), or lysine (alpha H45K). Moreover, alpha H44A and alpha H45A were distinct from the native luciferase in thermal stabilities. Mutations at the other three positions also resulted in activity reductions ranging from a fewfold to 3 orders of magnitude. Despite these widely different bioluminescence light outputs, mutated luciferases exhibited, in nonturnover in vitro assays, light emission decay rates mostly similar to that of the native luciferase using octanal, decanal, or dodecanal as a substrate. This is attributed to a similarity in the catalytic rate constants of the light-emitting pathway for the native and mutated luciferases, but various mutated luciferases suffer in different degrees from competing dark reaction(s). In accord with this interpretation, the bioluminescence activities of mutated luciferases showed a general parallel with the relative stabilities of their 4a-hydroperoxyflavin intermediate species. Furthermore, the drastically reduced bioluminescence activities for luciferases with the alpha His44 or alpha His45 substituted by aspartate, alanine, or lysine were accompanied by little or no activities for consuming the aldehyde substrate.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1991

Trimipramine: pharmacological reevaluation and comparison with clozapine.

Trimipramine has been reported to differ from other typical tricyclic antidepressant drugs in several aspects, for instance it does not inhibit neuronal transmitter uptake and does not cause down-regulation of beta-adrenoceptors. Moreover, it may possess antipsychotic activity in schizophrenic patients. In the present investigation it was found that trimipramine did not alter the electrically-induced release of [3H]noradrenaline and [3H]5-hydroxytryptamine, from slices of the cerebral cortex of the rat, in concentrations of less than 1 microM. It did not antagonize the inhibitory effect of noradrenaline and 5-hydroxytryptamine on the release of transmitter, mediated by presynaptic autoreceptors. In radioligand binding studies, D,L-trimipramine showed fairly high affinities (KI 10-60 nM) for some dopamine (DA), noradrenaline and 5-hydroxytryptamine (5-HT) receptor subtypes (5-HT2 receptors = alpha 1A/B-adrenoceptors greater than or equal to D2 receptors), intermediate affinities (300-550 nM) for D1 receptors, alpha 2B-adrenoceptors and 5-HT1C receptors but only low affinities (greater than 1000 nM) for alpha 2A-adrenoceptors, 5-HT1A, 5-HT1D and 5-HT3 receptors. It may thus be classified as an atypical neuroleptic drug. Especially, its affinities for dopamine receptors, alpha 1-adrenoceptors and 5-HT2 receptors closely resembled the values measured for clozapine. The L-enantiomer of trimipramine showed higher affinities for these binding sites than D-trimipramine. The present findings may explain the mechanism of the potential antipsychotic action but not the antidepressant effect of trimipramine.

Animals↗
PubMed1990

Urapidil analogues are potent ligands of the 5-HT1A receptor.

Urapidil and three derivatives with hypotensive properties (5-acetyl-, 5-formyl-, 5-methylurapidil) bind selectively to 5-HT receptors of the 5-HT1A subtype and to alpha 1-adrenoceptors labeled by [3H]8-OH-DPAT and [3H]prazosin, respectively. Binding to these receptors is likely to contribute to their hypotensive action. 5-Methylurapidil, the most potent of these drugs, was used in its 3H-labeled form as a radioligand. After blockade of alpha 1-adrenoceptors by prazosin, [3H]5-methylurapidil binds with nanomolar affinity to a binding site that is similar to the (5-HT1A) site labeled by [3H]8-OH-DPAT. No binding to other 5-HT1 and 5-HT2 receptors was observed. 5-HT uptake inhibitors did not inhibit [3H]5-methylurapidil binding. [3H]5-methylurapidil binding is sensitive to GTP and is modulated by divalent cations. Our results show that urapidil derivatives bind to the 5-HT1A recognition site and that [3H]5-methylurapidil is a valuable tool for the investigation of this receptor subtype.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1989

[Molecular cytogenetic study on the case with 14p+ marker chromosome].

A 23-year-old female was found carrying a submetacentric marker chromosome 14p+. The short arm of the marker chromosome showed a homogeneously stained region in both G-banded and C-banded preparations and was silver-positive in its distal part in AG-NOR preparation. Using a 3H-labelled 7.3 kb fragment of 18S and 28S rRNA genes as a probe and by chromosomal in situ hybridization, it was demonstrated that the autoradiographic silver grains to be located along entire p+ part of the marker chromosome. The short arm of the 14p+ had 5.3 times as many autoradiographic silver grains as that of any other normal acrocentric chromosome, but had no greater NOR activity than did the other acrocentric. The difference of these findings suggested that most of the amplified rRNA genes on the 14p+ chromosome were inactive, they might be inactivated by a process involving DNA methylation.

Adult↗
PubMed1986

Orally administrable brucellosis vaccine: Brucella suis strain 2 vaccine.

An orally administrable brucellosis vaccine, Brucella suis strain 2 vaccine was developed in China. The characteristics and merits of the vaccine are described. It is effective for oral vaccination of sheep, goats, cattle and pigs and has been widely used for prevention of animal brucellosis in China over the past 15 years. About 30-40 million doses of the vaccine are produced every year.

Administration, Oral↗
PubMedDate not supplied

Modeling pollutant release from a surface source during rainfall runoff.

Though runoff from manure spread fields is recognized as an important mode of nonpoint-source pollution, there are no models that mechanistically describe transport from a field-spread manure-type source. A mechanistic, physically based model for pollutant release from a surface source, such as field-spread manure, was hypothesized, laboratory tested, and field-applied. The primary objective of this study was to demonstrate the potential applicability of a mechanistic model to pollutant release from surface sources. The laboratory investigation used stable sources and a conservative "pollutant" (KCl) so that the dynamic effects of source dissolution and chemical transformations could be ignored and transport processes isolated. The field investigation used runoff and soluble reactive phosphorus (SP) data collected from a dairy-manure-spread field in the Cannonsville watershed in the Catskills region of New York State. The model predictions corroborated well with observations of runoff and pollutant delivery in both the laboratory and the field. "Pollutant" release from surface sources was generally predicted within 11% of laboratory KCl measurements and field SP observations. Laboratory flume runoff predictions with 15 and 26% errors for 25 and 15 mm h(-1) simulated rainfall intensity experiments, respectively, represented root mean square errors of less than 0.2 mLs(-1). A 26% error was calculated for overland flow predictions in the field, which translated into approximately a 39 mLs(-1) error. Results suggest that the hypothesized model satisfactorily represents the primary mechanisms in pollutant release from surface sources.

Agriculture↗