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Biomedical subjects

H Ding

Publications and source records attributed to H Ding.

At least 181 records · Page 10Linked to original sources

Hydroubiquinone-cytochrome c2 oxidoreductase from Rhodobacter capsulatus: definition of a minimal, functional isolated preparation.

The hydroubiquinone-cytochrome c2 oxidoreductase (cyt bc1) from Rhodobacter capsulatus has been solubilized according to the dodecyl maltoside method and isolated, and its minimal functional composition has been characterized. We find the complex to be composed of three protein subunits corresponding to polypeptides of cyt b (44 kDa), cyt c1 (33 kDa), and 2Fe2S cluster (24 kDa). A fourth band sometimes discernable at 22 kDa appears to be an artifact of the polyacrylamide gel electrophoresis procedure. Its appearance is shown to be derived from the 2Fe2S cluster subunit by the similarity of the binding of subunit-specific monoclonal antibodies and the identical N-terminal sequence of the 24- and 22-kDa bands. The cofactors of cyt bc1, namely, cyt bH, cyt bL, cyt c1, and the 2Fe2S center, the Qos and Qow domains of the Qo site, and the Qi site appear intact as indicated by their optical and EPR spectral signatures, redox properties, and inhibitor binding. The electron paramagnetic resonance spectrum of the cyt bH heme is altered by antimycin, consistent with a change in the dihedral angle between the ligating histidine imidazoles, while the spectrum of the cyt bL heme is broadened by stigmatellin. The ubiquinone-10 content is variable, ranging from 0.8 to 3 molecules/cyt bc1. Activity studies define this three-subunit cyt bc1 complex as a minimal structure, equipped as the enzyme in the native state and capable of full catalytic activity.

Amino Acid Sequence↗

Effect of intracerebroventricular injection of 6-hydroxydopamine on the peripheral catecholamine and angiotensin II in SHR.

This investigation revealed that the contents of A II and NE in plasma, heart and aorta and E content in plasma were significantly increased in SHR at the age of 12th week. At the same time the systolic blood pressure of SHR was much higher than that of the age-matched WKY. The intracerebroventricular administration of 6-OHDA in SHR at the age of 8th week not only prevented the development of hypertension, but also reduced the contents of NE and E in the brain regions, heart, aorta and plasma. Simultaneously A II content in heart, aorta and plasma was decreased. These results suggested that: 1. the renin-angiotensin system and peripheral sympatho-adrenal system are overactive in SHR, 2. the overactivity of peripheral sympatho-adrenal system is dependent on the central catecholaminergic neurons, and 3. the action of sympatho-adrenal system is partly responsible for the increase of A II content in plasma, heart and aorta in SHR.

Angiotensin II↗

Adherence of Actinomyces pyogenes to HeLa cells mediated by hydrophobic surface proteins.

Determination of cell-surface hydrophobicity of Actinomyces pyogenes by hydrophobic interaction chromatography on phenyl-Sepharose revealed that all 42 cultures examined were strongly hydrophobic. The hydrophobic surface proteins were solubilized by mutanolysin treatment of the bacteria and isolated by hydrophobic interaction chromatography. In SDS-PAGE, they appeared with numerous protein bands and blocked the adhesion of whole bacterial cells to the gel matrix. The A. pyogenes cultures attached to HeLa cells in varying degrees. This attachment of A. pyogenes was greatly reduced in the presence of isolated hydrophobic proteins and in the presence of specific antibodies produced against hydrophobic surface proteins. The results of the present study demonstrate that hydrophobic surface proteins promote the capacity of A. pyogenes to adhere to HeLa cells.

Actinomyces↗

Recombinant human insulin-like growth factor-I accelerates recovery and reduces catabolism in rats with ischemic acute renal failure.

This study evaluated whether recombinant human insulin-like growth factor-I (rhIGF-I) enhances recovery of renal function and reduces catabolism in rats with ischemic acute renal failure (ARF). ARF and sham rats received subcutaneous injections of either rhIGF-I or vehicle three times daily starting 5 h after surgery. Serum creatinine and urea, which initially rose similarly in the ARF+vehicle and ARF+rhIGF-I rats, increased more slowly after commencing the rhIGF-I injections. 72 h after surgery, the ARF+rhIGF-I rats, in comparison with ARF+vehicle animals, showed significantly greater renal plasma flow and filtration fraction, a fivefold higher glomerular filtration rate, greater renal cortical IGF-I levels, increased proliferating cell nuclear antigen expression in proximal tubule nuclei and enhanced DNA synthesis in the renal cortex, corticomedullary junction, glomeruli, and tubules as demonstrated by [3H]thymidine incorporation and in corticomedullary junction tubules as determined by autoradiography. Estimated total nitrogen output (ETNO) was greater in ARF+vehicle than in ARF+rhIGF-I or sham rats throughout the study. ETNO in ARF+rhIGF-I rats returned to sham values by the second day after surgery. 72 h after surgery, protein degradation was increased and protein synthesis reduced in the epitrochlearis muscle of ARF+vehicle as compared with ARF+rhIGF-I or sham+vehicle rats. Thus, treatment with rhIGF-I starting 5 h after inducing ischemic ARF in rats increases recovery of renal function, enhances formation of new renal tubular cells, lowers protein degradation, and increases protein synthesis in skeletal muscle and reduces net catabolism.

Acute Kidney Injury↗

[Cloning and sequencing of attacus ricini nuclear polyhedrosis virus polyhedrin gene].

Attacus ricini is a species of insect which only grows in China. It is expected to construct a new expression system for genetic engineering by using ArNPV as a vector. We have established an ArNPV gene library. The 1.1 kb DNA fragment containing ArNPV Ph gene was subcloned. Sequencing analysis shows that the 735 bp Ph structural gene has the homology of 76% and 81% with those of AcNPV and BmNPV respectively. The Rohrmann box in the ArNPV 5'-end regulation region is very similar to those of various other NPVs, but the 3'-end downstream sequence has almost no homology with those of AcNPV and BmNPV, demonstrating the characteristic property of the structure of ArNPV Ph gene. Other structural features of the Ph gene promoter are also discussed in this paper.

Base Sequence↗

In vitro inhibitory effect of IL-8 and other chemoattractants on neutrophil-endothelial adhesive interactions.

We have previously reported that cytokine- or LPS-activated human umbilical vein endothelial cell (HUVEC) monolayers secrete IL-8 that can act as a neutrophil-selective adhesion inhibitor. In our study we investigated the mechanisms involved in the leukocyte adhesion inhibitory action of IL-8. The leukocyte adhesion inhibitory effect appears to be mediated by the action of IL-8 on the neutrophil, does not involve down-regulation of relevant endothelial adhesion molecules such as endothelial-leukocyte adhesion molecule-1 or intercellular adhesion molecule-1, and is quantitatively similar in different endothelial activation states that are predominantly endothelial-leukocyte adhesion molecule-1 dependent or intercellular adhesion molecule-1 dependent. In addition to inhibiting the attachment of freshly isolated peripheral blood neutrophils to cytokine-activated HUVEC monolayers, IL-8 also promoted a rapid detachment of tightly adherent neutrophils from activated HUVEC, and abolished neutrophil transendothelial migration. Certain other chemoattractants, including FMLP and C5a, had similar inhibitory actions, indicating IL-8 was not unique in its ability to inhibit various neutrophil-endothelial interactions. In contrast, two other neutrophil agonists 1-0-alkyl-2-acetyl sn-glycero-3-phosphocholine and granulocyte-macrophage-CSF, which, like IL-8, are produced by activated HUVEC, as well as the leukocyte-derived chemoattractant leukotriene B4, exerted minimal inhibitory effects on adhesion. Regardless of their ability to modulate neutrophil-endothelial cell adhesion, all these agents induced altered leukocyte surface expression of functionally important adhesion molecules, including loss of L-selectin (leukocyte adhesion molecule-1, LECAM-1) and increase in CD11b/CD18. Thus, although the above agonists have been characterized primarily as chemoattractants, our findings demonstrate that these agents can exert a wide range of modulatory effects on neutrophil-endothelial adhesive interactions.

Antigens, CD↗

Cytochrome bc1 complex [2Fe-2S] cluster and its interaction with ubiquinone and ubihydroquinone at the Qo site: a double-occupancy Qo site model.

The ubiquinone complement of Rhodobacter capsulatus chromatophore membranes has been characterized by its isooctane solvent extractability and electrochemistry; we find that the main ubiquinone pool (Qpool) amounts to about 80% of the total ubiquinone and has an Em7 value close to 90 mV. To investigate the interactions of ubiquinone with the cyt bc1 complex, we have examined the distinctive EPR line shapes of the [2Fe-2S] cluster of the cyt bc1 complex when the Qpool-cyt bc1 complex interactions are modulated by changing the numbers of Q or QH2 present (by solvent extraction and reconstitution), by the exposure of the [2Fe-2S] to the Qpool in different redox states, by the presence of inhibitors specific for the Qo site (myxothiazol and stigmatellin) and Qi site (antimycin), and by site-specific mutations of side chains of the cyt b polypeptide (mutants F144L and F144G) previously identified as important for Qo site structure. Evidence suggests that the Qo site can accommodate two ubiquinone molecules. One (designated Qos) is bound relatively strongly and is second only to the ubiquinone of the QA site of the reaction center in its resistance to solvent extraction. In this strong interaction, the Qo site binds Q and QH2 with approximately equal affinities. Their bound states are distinguished by their effects on the [2Fe-2S] cluster spectral feature at gx at 1.783 (Q) and gx at 1.777 (QH2); titration of the line-shape change reveals an Em7 value of approximately 95 mV. The other molecule (Qow) is bound more weakly, in the same range as the ubiquinone of the QB site of the reaction center. Again, the affinities of the Q form (gx at 1.800) and QH2 form (gx at 1.777) are nearly equal, and the Em7 value measured is approximately 80 mV. These results are discussed in terms of earlier EPR analyses of the cyt bc1 complexes of other systems. A Qo site double-occupancy model is considered that builds on the previous model based on Qo site mutants [Robertson, D. E., Daldal, F.,& Dutton, P. L. (1990) Biochemistry 29, 11249-11260] and includes the recent suggestion that two of the [2F3-2S] cluster ligands of the R. capsulatus cyt bc1 complex are histidines [Gurbiel, R. J. Ohnishi, T., Robertson, D. E. Daldal, F., & Hoffman, B. M. (1991) Biochemistry 30, 11579-11584]. We speculate that the cyt bc1 complex complexes a full enzymatic turnover without necessary exchange of ubiquinone with the Qpool.

Bacterial Chromatophores↗

Cell surface hydrophobicity of Actinomyces pyogenes determined by hexadecane adherence- and salt aggregation studies.

Cell surface hydrophobicities of Actinomyces pyogenes were determined by measuring the adherence of the bacteria to hexadecane droplets and by salt aggregation tests. Among 42 A. pyogenes cultures tested 25 (60%) adhered strongly (adherence greater than or equal to 75%) and 17 (40%) less pronounced (adherence between 25-75%) to the hexadecane droplets. Pre-treatment of the bacteria with proteolytic enzymes completely eliminated the adherence properties whereas heat treatment had no effect. The salt aggregation studies revealed that 4 (10%) cultures aggregated in ammonium sulfate solutions of a molarity of 0.05 mol/l, 5 (12%), 14 (33%) and 3 (7%) cultures in ammonium sulfate solutions with molarities of greater than or equal to 1.5 mol/l, greater than or equal to 3 mol/l and greater than or equal to 4.5 mol/l, respectively. No aggregation at all could be observed with 16 (38%) of the cultures. Pronase treatment completely eliminated the salt aggregation reactions, trypsin- and heat treatment had no effect. The results from hexadecane adherence and salt aggregation did not correspond. The differences in surface hydrophobicities, possibly related to adherence properties of A. pyogenes, could be used for epidemiological typing of individual cultures of this bacterial species.

Actinomyces↗

Evaluation of the API Coryne test system for identification of Actinomyces pyogenes.

The present study was designed to evaluate the accuracy of the API Coryne test system for identification of Actinomyces pyogenes. The test system correctly identified 36 of 42 A. pyogenes and 4 of 5 comparatively studied Arcanobacterium haemolyticum-cultures. The biochemical profiles of the remaining 6 A. pyogenes- and 1 A. haemolyticum-cultures were not included in the analytical profile index. None of the cultures were misidentified. According to the API database (ATB Plus V 1.5.4.) the unidentified cultures could be correctly identified as A. pyogenes and A. haemolyticum respectively. A greater repertoire of A. pyogenes specific biochemical profiles incorporated into the analytical profile index would improve the applicability of this test system for veterinary diagnostics.

Actinomyces↗

[Measurement of dopamine in rat striatum in vivo with a biomicroelectrode made from mixed plant tissue-carbon paste].

The mixed plant tissue-carbon paste electrode was prepared and their electrochemical characteristics were investigated. The sensitivity and selectivity of this bioelectrode were found to be good because of the utility in biocatalysis. It may work continuously with high stability for 10 h in vivo. The principal advantages of the new bioelectrodes are shorter in response time and longer in stability. These advantages meet the requirements of in vivo determination. Using these electrodes, the dopamine contents in rat striatum were measured by anodic stripping voltammetry. The drug-induced changes in dopamine levels were monitored. From the results, we can conclude that the biomicroelectrode is a very economic biosensor for its low cost and easiness to prepare and is a valuable tool for studying dopamine function. It will provide a good method for the evaluation of drug actions on dopamine neurones.

Animals↗

[Hepatic segmentectomy using microwave tissue coagulator].

Hepatic segmentectomy using a microwave tissue coagulator guided by intraoperative ultrasonography is a new operative procedure, which our research unit was the first to start using from 1990. Up to now we have performed this kind of operation with success in 26 cases. Our results suggested that the new procedure simplified the original operation and greatly reduced the risk of hemorrhage and iatrogenic spread of the cancer cells during operation. Besides, this operation as a kind of definite anatomic hepatectomy can minimally resect the tumor bearing tissue in a radical fashion, while maximally preserve the tumor-free tissue of the liver.

Adult↗

[Central norepinephrine and angiotensin II contents in the brain regions of spontaneously hypertensive rats (SHR) and the interaction between them].

The norepinephrine (NE) and angiotensin II (A II) contents in the brain regions of SHR and WKY (Wistar Kyoto) rats at different ages were determined by fluorospectrophotometry and radioimmunoassay. The systolic blood pressure (SBP) of the rats was measured indirectly with a tail cuff technique in conscious state. The results were as follows: There was no significant difference in the central A II and NE contents between SHR and WKY rats at 8-week age. Since 12th week age the SBP of SHR has increased gradually, up to 16th to 20th week and then maintained steady level. Whereas there was no significant change of SBP in WKY rats in the same span of age. In the early and late states of hypertension the A II contents in the medulla oblongata, pons, hypothalamus and nucleus caudatus of SHR were markedly higher than those of the age-matched WKY rats. But the change of NE content of SHR in the early stage showed a different picture as compared with that of WKY rats, i.e., NE decreased in medulla oblongata and anterior hypothalamus but increased in pons, posterior hypothalamus and nucleus caudatus. However, in the late stage there was no such significant difference between SHR and WKY rats. Consequently, it is suggested that the central A II and NE participated in the development of hypertension of SHR, and that the maintenance of hypertension is mainly dependent upon the increased A II content. Microinjection of captopril or 6-OHDA in the lateral cerebroventricle of SHR elicited a decrease of BP and reduction of both A II and NE contents in the medulla and hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗