Search PubMed⌕ Search

Biomedical subjects

M Wikström

Publications and source records attributed to M Wikström.

At least 181 records · Page 10Linked to original sources

Postnatal development of catecholamines and response to hypoxia in adrenals and paraganglia of rabbits.

The postnatal development of catecholamine levels in adrenals and paraganglia in newborn rabbits has been analyzed. At birth, the dominant catecholamine was noradrenaline, contributing 66% of the total catecholamine pool at day 1, 55% of which came from the paraganglia. There was a rapid postnatal increase of adrenaline, which constituted 67% of the total catecholamines at day 6 and 97% in the adult. After hypoxia at day 1, the noradrenaline levels decreased in paraganglia but not in adrenals, while adrenaline levels did not significantly change in either organ.

Adrenal Glands↗

Structural models of the redox centres in cytochrome oxidase.

Evolutionary conservation, predicted membrane topography of the subunits, and known chemical and physical properties of the catalytic metals in cytochrome oxidase provided the basis for plausible structural models of the enzyme's redox centres. Subunit II probably binds one of the copper ions (CuA) whilst subunit I is likely to bind the two haems (a and a3) and the other redox-active copper (CuB). Two cysteine and two histidine residues of subunit II are the likely ligands of CuA, forming a centre that may be structurally similar to that in azurin. The two haems may be sandwiched between two transmembranous segments of subunit I, one of which also provides a histidine ligand to CuB. A third segment may provide two more histidine ligands to the latter. The model was constructed with a 4 A Fe-Cu distance in the binuclear haem a3-CuB centre, and a 14 A distance between the haem irons. The subunit I model involves only three transmembranous helices which bind three catalytic metal groups. The fit of this model to several known physicochemical properties of the redox centres is analysed.

Animals↗

Principles of coupling between electron transfer and proton translocation with special reference to proton-translocation mechanisms in cytochrome oxidase.

The recent general acceptance of the proton-pumping function of cytochrome oxidase has stimulated discussion and experiment on possible underlying molecular mechanisms. Adequate experimental design requires clear understanding of the theoretical principles governing such a linked function. The increasing structural knowledge of cytochrome oxidase also contributes to a present-day requirement of more precise chemical and physical description of redox-linked proton translocation, which is the fundamental process underlying conservation of energy from aerobic metabolism in all eukaryotes and many bacteria. This essay is based on our original theoretical treatment of this problem, which is expanded here to include discussion of more recent analyses by others, classification of different types of coupling principles, as well as some concrete proposed molecular mechanisms. The latter will be analysed qualitatively, and in some cases quantitatively where this is possible, using a common theoretical framework to help comparison between models. Experimental findings relevant to this problem will be critically reviewed, and some suggestions will be made to stimulate further experiments dedicated to clarify the problem.

Biological Transport↗

The capability of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius to indicate progressive periodontitis; a retrospective study.

This study evaluated the statistical association of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius with progressive periodontitis. 146 adults with a history of advanced periodontitis contributed 105 "nonprogressing" and 130 "progressing" periodontal sites. Periodontal disease activity was assessed by radiographic changes in crestal alveolar bone level. The subgingival proportion of the 3 test bacteria was determined by selective and nonselective culturing. The relationship between bacterial proportions and disease progression was evaluated using subgrouping and multiple-regression analyses. All 3 test bacteria had to be considered in order to distinguish nonprogressing and progressing periodontitis with a reasonably high sensitivity. A recovery rate below 0.01% for A. actinomycetemcomitans, 0.1% for B. gingivalis and 2.5% for B. intermedius defined a site with nonprogressing disease with 87% sensitivity and 84% specificity. By utilizing transformed values of the bacterial recovery rates and optimal test criteria determined by multiple regression analysis, it was possible to obtain sensitivities between 83% and 95% and specificities between 86% and 69%. These 3 bacterial species might serve as valuable components of a periodontitis activity test based on microbiological variables.

Actinobacillus↗

Studies on the role of the oligomeric state and subunit III of cytochrome oxidase in proton translocation.

Anion-exchange fast protein liquid chromatography in the presence of lauryldimethylamine N-oxide (LDAO) was introduced to separate cytochrome oxidase into different complexes that either did or did not contain subunit III. Both kinds of enzyme complex exhibited H+ translocation after reconstitution into phospholipid vesicles, but with a significantly (approx. 50-60%) reduced H+/e- ratio as compared with unchromatographed enzyme. The anion-exchange FPLC fractions of the enzyme (with or without subunit III) sedimented more slowly than the control enzyme upon sucrose gradient centrifugation in the presence of cholate and a high potassium phosphate concentration. When the control enzyme was subjected to the sucrose gradient centrifugation in the presence of LDAO or Triton X-100, instead of cholate, one band containing all subunits was observed, which sedimented slowly like the FPLC fractions. Transfer of this band to cholate medium, and reapplication on the sucrose gradient (with cholate), yielded both a slow- and a fast-migrating band after centrifugation. Enzyme complexes that sedimented slowly or rapidly in the sucrose gradients revealed longer and shorter elution times, respectively, in gel filtration FPLC. This suggests that these complexes corresponds to monomers and dimers of cytochrome oxidase. Solubilization of proteoliposomes and subsequent sucrose gradient centrifugation in cholate yielded one fast-migrating band for the untreated enzyme, but both a fast- and a slow-migrating band for the anion-exchange FPLC-treated enzyme, which was exclusively slow-migrating before reconstitution into liposomes. It is suggested that dimerisation of monomeric cytochrome oxidase may be favoured when the enzyme encounters a membranous milieu, and that the dimeric structure might be necessary for proton translocation.

Animals↗

Determination of the stoichiometry of redox-linked proton translocation from the kinetics of pulse experiments. A simulation study.

We have previously published a simple kinetic model to analyse possible pitfalls in kinetic measurements of H+/O ratios in mitochondria [(1984) FEBS Lett. 178, 187-192]. While this model demonstrated how relative electrode response times may affect the results, it did not adequately describe the kinetics of proton back-diffusion across the membrane. Here this model is further developed and improved, and shown to give a good quantitative description of both oxygen-pulse type experiments as well as of experiments where the reaction is started by photolysis of the cytochrome c oxidase-CO complex. Simulations based on this model reveal that the extrapolation procedure used by Lehninger et al. [e.g. (1984) J. Biol. Chem. 259, 4802-4811] to estimate the H+/O ratio will tend to yield overestimated values. This is mainly due to the back-diffusion of protons into the mitochondria, which is not correctly accounted for by this extrapolation.

Carbon Monoxide↗

Evidence for a mobile semiquinone in the redox cycle of the mammalian cytochrome bc1 complex.

Experimental evidence is presented to demonstrate that cytochromes b of the mammalian cytochrome bc1 complex may be rapidly oxidised by a pulse of oxidising equivalents which react with cytochrome c1, even when all cytochrome b is fully reduced before the pulse. The oxidation is sensitive both to antimycin and to myxothiazol. Such behaviour is inconsistent with models in which only the fully oxidised ubiquinone may move between the centres 'o' and 'i' of the complex. It is proposed that the charged semiquinone (Q-) may move between these centres, which may constitute separate reaction domains of a single ubiquinone-binding site. The bearing of this on the mechanism of electron, proton and charge transfer in the complex is discussed.

Animals↗

The semiquinone cycle. A hypothesis of electron transfer and proton translocation in cytochrome bc-type complexes.

The Q cycle and the b cycle are the main current models of action of the cytochrome bc-type complexes of mitochondria, bacteria, and chloroplasts. Both are based on the concept, proposed in 1972, of two sequential one-electron oxidations of (ubi)quinol along two discrete pathways which operate at different redox potentials, and with bound semiubiquinone as an intermediate. The models differ in two respects, viz. in the pathway of electron transfer and the principle of linkage of electron transfer to proton translocation. In this article we outline a new model, called the semiquinone or, simply, SQ cycle, which is based on the electron transfer principles of the b cycle but which incorporates the Q cycle concept of direct coupling between electron transfer and proton translocation through action of ubiquinone.

Benzoquinones↗

The occurrence of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius in destructive periodontal disease in adults.

A total of 235 subgingival sites, including 104 progressive deep lesions from 61 untreated patients, 26 progressive deep lesions from 10 treated patients, 33 nonprogressive deep sites from 20 untreated patients, and 72 nonprogressive sites from 55 treated patients were examined for Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius. The periodontal disease progression was mainly determined on the basis of radiographic changes in the crestal alveolar bone level. A. actinomycetemcomitans isolation was carried out using the selective TSBV medium and B. gingivalis and B. intermedius isolations were performed using a nonselective blood agar medium. 1 or more of the 3 bacteria studied appeared in 99.2% of progressive periodontal lesions but only in 40.0% of nonprogressive sites. Culture-positive progressive periodontal sites in comparison with culture-positive nonprogressive sites showed higher median recovery rates of A. actinomycetemcomitans (0.5% vs 0.3%), B. gingivalis (30.5% vs 0.3%) and B. intermedius (4.9% vs 0.5%). Of total progressive lesions, 12.3% yielded solely A. actinomycetemcomitans, 21.5% demonstrated solely B. gingivalis, and 20.8% revealed solely B. intermedius. The A. actinomycetemcomitans--B. intermedius combination was found in 24.6% of progressive lesions. A. actinomycetemcomitans appeared in significantly higher prevalence in treated-progressive lesions (80.8%) than in nontreated-progressive lesions (42.3%). 32 of the 42 culture-positive nonprogressive sites yielded B. intermedius as the sole test organism. The main conclusion is that A. actinomycetemcomitans, B. gingivalis and B. intermedius are closely related to disease-active periodontitis, and more closely than to periodontal pocket depth. This finding is important in understanding periodontal disease etiology and pathogenesis and may also aid in a clinical setting to differentiate progressing and nonprogressing periodontal sites.

Actinobacillus↗

Ability of oral bacteria to degrade fibronectin.

The fibronectin-degrading ability of 116, mainly oral, strains was assayed by using plasma-derived fibronectin adsorbed to a polystyrene surface. Ability to degrade fibronectin was revealed in strains of Bacteroides gingivalis, Bacteroides intermedius, Bacteroides loeschii, Staphylococcus aureus, Staphylococcus epidermidis, Peptococcus prevotii, Clostridium sporogenes, and Propionibacterium acnes. The fibronectinolytic activity of subgingival bacteriological samples was found to be related to the presence of B. gingivalis and B. intermedius. In addition, strains of the nonoral Bacteroides species B. asaccharolyticus and B. fragilis showed fibronectin-degrading ability. No such ability was detected in the oral strains tested of Streptococcus, Veillonella, Actinomyces, Lactobacillus, Actinobacillus, Capnocytophaga, Fusobacterium, or Haemophilus species.

Bacteria↗

Monoclonal antibodies against Escherichia coli heat-stable toxin (STa) and their use in a diagnostic ST ganglioside GM1-enzyme-linked immunosorbent assay.

Seven monoclonal antibodies (MAbs) against heat-stable enterotoxin (ST) from a human Escherichia coli isolate were prepared and evaluated for their usefulness in an ST immunodetection assay, the ST ganglioside GM1-enzyme-linked immunosorbent assay (ELISA). This assay is based on the ability of STa, as present in, for example, culture filtrates from ST-producing E. coli, to inhibit specific anti-ST antibody from binding to solid-phase-bound ST ganglioside (GM1-bound ST-cholera B subunit). Four of the MAbs were of immunoglobulin G1 (IgG1), one was of IgG2b, and two were of IgM isotype. All the IgG1 MAbs could be completely inhibited by addition of free ST; 0.2 to 0.4 ng of purified ST inhibited binding of these MAbs by 50%. The non-IgG1 MAbs were, in contrast, not inhibited by 200-fold-higher amounts of purified ST, probably because they were directed against linkage epitopes or were of low affinity or both. When the IgG1 MAbs were tested in the ST GM1-ELISA, ST could be detected in culture filtrates from stock human E. coli isolates with 100% sensitivity and specificity. ST in filtrates from fresh stool cultures was demonstrated with higher sensitivity with the MAbs ST GM1-ELISA than with the conventional infant mouse test. Both subtypes of STa, STaI and STaII, could be detected by the ST GM1-ELISA by using either IgG1 MAb in the immunodetection step, whereas infant-mouse-active ST from Yersinia enterocolitica failed to react.

Animals↗

Monoclonal antibodies to Escherichia coli heat-labile enterotoxins: neutralising activity and differentiation of human and porcine LTs and cholera toxin.

Forty-four monoclonal antibodies (MAbs) prepared against heat-labile enterotoxins (LTs) from human (LTh) or porcine (LTp) E. coli isolates were characterised, especially with regard to their reactivity with epitopes shared with the heterologous LT and/or cholera toxin (CT), and their toxin neutralising activity. Of 24 MAbs against LTh (all directed against the B subunit portion) 12 cross-reacted with LTp and CT, 4 with LTp but not CT, and 1 with CT but not LTp; 7 MAbs reacted with LTh epitope(s) not shared by either LTp or CT. Among 20 MAbs against LTp (9 directed against the B subunits and 11 against the A subunit) 2 cross-reacted with LTh as well as CT, 13 with LTh but not CT, and 5 MAbs were specific for LTp. Irrespective of whether the anti-LT MAbs were directed against shared or unshared epitopes, or against the A or B subunits, they neutralised their homologous toxin in direct proportion to their toxin-binding titre. The results show how minute differences in enterotoxin primary structures e.g., the LTh and LTp B chains differ in only 4 of 103 amino acid residues, are associated with antigenic epitopes against which toxin-differentiating MAbs with neutralising activity can be produced. Such MAbs are promising tools for species-specific diagnostic detection of enterotoxins in clinical specimens.

Amino Acids↗

The oxidation of exogenous cytochrome c by mitochondria. Resolution of a long-standing controversy.

Several reports in the past have dealt with the oxidation of cytochrome c added to suspensions of rat liver mitochondria. Yet, it is generally believed that the cytochrome cannot penetrate the outer membrane. Probably it has been assumed that the permeability of the outer membrane to cytochrome c is very low but finite, and that fast oxidation may be observed if time is allowed for sufficient penetration before initiation of electron flow. Here we show that this view is false. The main fraction of rat liver mitochondria, as isolated by conventional procedures, does not catalyse any significant oxidation of added cytochrome c, even after prolonged incubation. The observed appreciable oxidation of added cytochrome c is catalysed by a very small fraction (5-12%) of the mitochondria that apparently has a damaged outer membrane. Consequently, the turnover of cytochrome oxidase is very high in this fraction during oxidation of added cytochrome c. This finding readily explains why Moyle and Mitchell (e.g., FEBS Lett. 88 (1978) 268-272; 90 (1978) 361-365) have failed to observe proton translocation by cytochrome oxidase during oxidation of ferrocytochrome c added to rat liver mitochondria, which has been their main reason for rejecting the proton-pumping function of cytochrome oxidase.

Animals↗

Retinoids activate superoxide production by polymorphonuclear leucocytes.

Retinol and retinoic acid were effective activators of oxygen consumption by human polymorphonuclear leucocytes (PMN) in micromolar concentrations. In contrast, retinyl acetate was ineffective as an activator. Retinol caused activation only after a lag time, the length of which depended on retinol concentration. Oxygen consumption was due to superoxide production by PMN. Superoxide production was observed as superoxide dismutase-inhibitable cytochrome c reduction. Previously, retinoids have been reported to inhibit PMN activation by phorbol myristate acetate, a tumour promoter. This retinoid-induced inhibition of PMN activation has been suggested to be a mechanism by which retinoids may protect against carcinogenesis in animals. However, the retinoid concentrations at which PMN inhibition was reported were much higher than those found to cause activation in this study. We found that retinoic acid slightly inhibited phorbol myristate acetate-activated superoxide production, but only at concentrations that caused activation. In contrast, activation by formyl-Met-Leu-Phe was effectively inhibited at a retinoic acid concentration that did not cause activation by itself.

Diterpenes↗

The H+/O ratio of proton translocation linked to the oxidation of succinate by mitochondria.

In a recent communication Lehninger and co-workers (Costa, L.E., Reynaferje, B., and Lehninger, A.L. (1984) J. Biol. Chem. 259, 4802-4811) reported values approaching 8 for the H+/O ratio of vectorial proton ejection from rat liver mitochondria respiring with succinate. Here we present a rigorous analysis of these measurements which reveals that they may significantly overestimate the true H+/O stoicheiometry.

Animals↗