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Biomedical subjects

A Kemp

Publications and source records attributed to A Kemp.

At least 127 records · Page 7Linked to original sources
PubMed1981

Photolabelling with 8-azido-adenine nucleotides of adenine nucleotide-binding sites in isolated spinach chloroplast ATPase (CF1).

1. Photolabelling of chloroplast ATPase (CF1) with either 8-azido-ATP or 8-azido-ADP leads to inactivation of the ATPase activity. ATP and ADP protect against the inactivation, whereas AMP dose not. 2. Ca2+ has little if any effect on the degree of inactivation by photolabelling with 8-azido-ADP, but, at the same degree of inactivation, twice as much label is bound in the presence of Ca2+ as in its absence. 3. The degree of inactivation of ATPase and the amount of bound photolabel are independent of the extent of pre-activation of the CF1. 4. Upon extrapolation to complete inactivation, 2 mol label, either 8-azido-ATP or 8-azido-ADP can be bound. 5. In all cases the label is bound specifically to the alpha and beta subunits in almost equal amounts. The location of the bound label is not affected by addition of Ca2+, ATP or ADP.

Adenosine Diphosphate↗
PubMed1981

The function of ascorbate with respect to prolyl 4-hydroxylase activity.

1. Incubation in the presence of 2-oxoglutarate and oxygen inactivates prolyl 4-hydroxylase (prolyl-glycyl-peptide, 2-oxoglutarate:oxygen oxidoreductase, EC 1.14.11.2), with a t 1/2 of 80 s at 37 degrees C. This inactivation is not affected by the presence or absence of the prolyl peptide substrate or added Fe(II). 2. This inactivation can be prevented by either ascorbate or dithiothreitol. It can be reversed by dithiothreitol but not by ascorbate. 3. Although the iron-containing form of prolyl 4-hydroxylase requires ascorbate for activity, ascorbate is not stoicheiometrically consumed in the reaction catalysed by the enzyme. Ascorbate cannot be replaced by alloxan, lactate, NADH plus phenazine methosulphate, dithiothreitol or L-cysteine. 4. Ascorbate has a double function with respect to prolyl 4-hydroxylase activity. On the one hand, it is required to initiate the reaction when the enzyme has become oxidized during isolation. On the other hand it is required for the protection against inactivation induced by 2-oxoglutarate and oxygen, presumably by preventing S-S bridge formation. The latter function may be of physiological importance.

Animals↗
PubMed1980

The number and localisation of adenine nucleotide-binding sites in beef-heart mitochondrial ATPase (F1) determined by photolabelling with 8-azido-ATP and 8-azido-ADP.

1. When irradiated 8-azido-ATP becomes covalently bound (as the nitreno compound) to beef-heart mitochondrial ATPase (F1) as the triphosphate, either in the absence or presence of Mg2+, label covalently bound is not hydrolysed. 2. In the presence of Mg2+ the nitreno-ATP is bound to both the alpha and beta subunits, mainly (63%) to the alpha subunits. 3. After successive photolabelling of F1 with 8-azido-ATP (no Mg2+) and 8-azido-ADP (with Mg2+) 4 mol label is bound to F1, 2 mol to the alpha and 2 mol to the beta subunits. 4. When the order of photolabelling is reversed, much less 8-nitreno-ATP is bound to F1 previously labelled with 8-nitreno-ADP. It is concluded that binding to the alpha-subunits hinders binding to the beta subunits. 5. F1 that has been photolabelled with up to 4 mol label still contains 2 mol firmly bound adenine nucleotides per mol F1. 6. It is concluded that at least 6 sites for adenine nucleotides are present in isolated F1.

Adenine Nucleotides↗
PubMed1980

Nitrate poisoning in cattle. 4. The effect of nitrite dosing during parturition on the oxygen capacity of maternal blood and the oxygen supply to the unborn calf.

In a series of experiments the effect of administering KNO2 was studied, during parturition, on the capability of oxygen transport of maternal blood and on oxygen transfer to foetal blood. The following blood parameters were analysed, MHb percentage, pO2, O2-saturation, pH, pCO2, and (NO2) in maternal arterial blood (carotid art.) and venous blood (jugular vein) and in foetal arterial blood (umbilical art.) and venous blood (umbilical vein). The relative O2-saturation was calculated from the estimated O2-saturation by multiplying with the factor Hb (mmol/l) minus MHb (mmol/l), divided by Hb (mmol/l). In addition, blood pressure in the carotid artery, heart rate, and respiration rate in the dam were continuously recorded for some hours. A dosage of 9 to 12 mg of NO1/kg body weight intravenously or of 30 mg of NO2/kg body weight orally to the dam caused much higher MHb percentages and NO2 contents in the maternal blood than in the foetal blood. In maternal blood the ratio of NO2 content to MHb percentage was proportional to that in foetal blood. In the arterial blood, MHb percentages were almost as high as in the venous blood. After administering of nitrite, relative O2-saturation dropped simultaneously with the increase in methaemoglobin. Nitrite treatment caused a drop in the maternal blood pressure; heart rate and respiration rate increased. O2-saturation in the blood in the umbilical vein was much lower in the animals with nitrite treatment than in those without. These experimental results show clearly that the oxygen capacity of the blood decreases after nitrite treatment. In pregnant cows the oxygen supply to the foetus will be adversely affected after nitrate intake, especially by the lower oxygen transfer via the placenta, though hardly at all by methaemoglobin formation in the foetal blood. When the oxygen transfer to the foetal blood decreases too sharply, intra-uterine death and ultimately abortion may result.

Animals↗
PubMed1980

Properties of prolyl 4-hydroxylase containing firmly-bound iron.

1. Prolyl 4-hydroxylase (prolyl-glycyl-peptide, 2-oxoglutarate:oxygen oxidoreductase, EC 1.14.11.2) was isolated in a form containing iron (0.85-1.1 mol Fe/mol enzyme). 2. The enzyme was pure according to gel electrophoresis and had a high specific activity (1.8-2.6 mumol . mi-1 . mg-1). 3. Experiments with metal chelators showed this iron to be firmly bound and to be required for catalytic activity. 4. According to EPR spectrometry the bound iron is not part of a [2Fe-2S] or a [4Fe-4S] cluster. 5. The enzyme activity is to a large extent independent of added Fe2+. 6. The enzyme activity is completely dependent on ascorbate. 7. In the absence of ascorbate but the presence of substrates the enzyme is irreversibly inactivated. 8. Continuous measurement of enzyme activity was possible by following oxygen uptake.

Animals↗
PubMed1980

EEG observations in dystrophia myotonica (Curschmann-Steinert).

An investigation has been carried out in a group of 84 (+2) patients suffering from myotonic dystrophia (Curschmann-Steinert). Manual measurements show that in these patients: (1) the alpha rhythms have low frequencies, related to the age of the patient and to the duration and severity of the disease; (2) the alpha rhythms are regular, monotonous; (3) the alpha rhythms have small amplitudes; there appears to be no relation between alpha frequency and amplitude; (4) the alpha indices are high during eyes closed as well as during eyes open. The computer measurements generally appear to concur with the above and in addition show that the bilateral and longitudinal coherences are high, indicating considerable synchrony of the alpha rhythm over large areas on both sides of the head. The possible significance of these observations is discussed.

Adolescent↗
PubMed1980

HLA antigens in cases of giant cell arteritis.

HLA tissue-type antigen determination for A-, B- and C-antigens in 88 patients suffering from giant cell arteritis of the temporal artery showed no significant deviations as compared to a control material of 3164 blood donors. A weak indication of association with antigen HLA-B8 appeared to be of interest due to a corresponding indication in previous investigation. The patients were a mixed hospital material, consisting of cases of clinical temporal arteritis and patients with polymyalgia rheumatica. There was an overrepresentation of women (77%). Familial occurrence was demonstrated sporadically (3 pairs of siblings).

Aged↗
PubMed1979

Localisation of adenine nucleotide-binding sites on beef-heart mitochondrial ATPase by photolabelling with 8-azido-ADP and 8-azido-ATP.

1. In addition to the previously studied 8-azido-ATP, 8-azido-ADP is a suitable photoaffinity label for beef-heart mitochondrial ATPase (F1). 2. Photolysis at 350 nm of 8-azido-ADP in the presence of isolated F1 leads to inactivation of ATPase activity. Both ATP and ADP (but not AMP) protect against the inactivation. 3. In the absence of Mg2+, 8-azido-ADP binds almost equally to the alpha and beta subunits of F1, whereas in the presence of Mg2+ the alpha subunits are predominantly labelled. 4. The ATPase activity is completely inhibited when two molecules of 8-azido-ADP are bound per molecule F1. 5. 8-Azido-ATP and ATP are competitive substrates for F1, indicating that in the presence of Mg2+ 8-azido-ATP binds to the same site as ATP. 6. The amount of tightly bound nucleotides in F1 is not significantly changed upon incubation with 8-azido-ATP either in the light or the dark. 7. 8-Azido-ATP is also a suitadrial particles, photolabelling leading to inactivation of ATPase activity. 9. Oxidative phosphorylation and the ATP-driven reduction of NAD+ by succinate are also inhibited by photolabelling Mg-ATP particles with 8-azido-ATP. 10. In contrast to the uncoupled ATPase activity, where the two ATP-binding sites do not interact, cooperation between the two sites is required for ATP hydrolysis coupled to reduction of NAD+ by succinate.

Adenosine Diphosphate↗
PubMed1979

Inhibition of human neutrophil migration by aggregated gammaglobulin.

Heat-aggregated human gammaglobulin has been shown to inhibit the random migration of human neutrophils in serum-containing medium. This inhibition was not due to metabolic exhaustion or deactivation of the cells, since migration in the presence of aggregated gammaglobulin and casein as a chemotactic stimulus was not inhibited. The inhibition of migration was not mediated by a negative chemotactic gradient produced as a result of complement activation, and could be demonstrated in complement-depleted serum. Sera obtained from patients with rheumatoid arthritis with evidence of circulating immune complexes were able to significantly inhibit neutrophil migration, indicating that this phenomenon may be a useful means for the detection of circulating immune complexes. It is suggested that aggregated gammaglobulin or immune complexes can inhibit the chemokinetic effect of serum on neutrophils by a reversible interaction with the neutrophil surface, and that this inhibition could contribute to the accumulation of neutrophils at sites of immune complex deposition in vivo.

Adult↗
PubMed1978

Brown-adipose-tissue mitochondria: photoaffinity labelling of the regulatory site of energy dissipation.

Brown-adipose-tissue mitochondria possess an energy-dissipating ion uniport which is inhibited by purine nucleotides. The regulatory nucleotides bind to a high-affinity site on the outer face of the inner membrane which is independent of the adenine nucleotide translocator. A direct correlation between affinity for the regulatory site and ability to inhibit the ion uniport is demonstrated for a number of nucleotide analogues. 8-Azido-adenosine 5'-triphosphate, a photoaffinity label, also competes with GDP for the binding site and induces respiratory control. 8-Azido-adenosine [gamma-32P]triphosphate was prepared and covalently bound to hamster brown-adipose-tissue mitochondria by near-ultraviolet irradiation. Two major radioactive bands were identified of apparent molecular weight 30000 and 32000, representing 6% and 10% of the inner membrane protein respectively. Selective labelling enabled the 30000-Mr protein to be identified as the carboxyatractylate binding component of the adenine-nucleotide translocator and the 32000-Mr protein to be identified as the regulatory site of the energy-dissipating ion uniport. The levels of the 32000-Mr protein in the inner membrane of guinea-pig brown-adipose-tissue mitochondria correlate with the degree of thermogenic adaptation of the animal.

Adenosine Triphosphate↗
PubMed1977

Specific photolabelling of beef-heart mitochondrial ATPase by 8-azido-ATP.

1. 8-Azido-ATP is a suitable photoaffinity label for beef-heart mitochondrial ATPase (F1) 2. 8-Azido-ATP is hydrolysed slowly by F1 in the dark. Photolysis at 350 nm in the presence of F1 leads to inhibition of the ATPase activity. The presence of ATP during illumination prevents the inhibition. Illumination of F1 in the absence of 8-azido-ATP causes no inhibition. 3. Added Mg2+ is not necessary for the binding of the 8-azido-ATP to F1. 4. 8-Azido-ATP binds specifically to the beta subunits of F1. 5. The ATPase activity is completely inhibited when 2 mol of 8-azido-ATP are bound per mol F1.

Adenosine Triphosphatases↗
PubMed1977

Uncoupling of mitochondrial respiration by ADP.

Even when oxidative phosphorylation is blocked completely by addition of high concentrations of oligomycin plus aurovertin, the addition of ADP to a suspension of mitochondria containing a high concentration of ATP inside the mitochondria induces a stimulation of respiration and oxidation of nicotinamide nucleotide. It is concluded that transport of ADP into mitochondria with a high endogenous ATP/ADP ratio requires energy.

Adenosine Diphosphate↗
PubMed1977

Monozygotic twins with temporal arteritis and ophthalmic arteritis.

Temporal arteritis - ophthalmic arteritis occurring in a pair of monozygotic twins, who were 72 and 77 years of age respectively at the onset of symptoms, is reported. It is pointed out that it is rare to see, as in the first case, an improvement in an already existing severely reduced vision, in relation to corticosteroid treatment. The importance of genetic factors in temporal arteritis is discussed; further it is suggested that a possible association of the disease with tissue type antigens be the object of further study.

Aged↗
PubMed1976

Role of the intramitochondrial adenine nucleotides as intermediates in the uncoupler-induced hydrolysis of extramitochondrial ATP.

1. A formula is given that describes the appearance of [14C]ATPADP outside the mitochondria after the addition of [14C] 1atp during the steady-state uncoupler-induced hydrolysis of extramitochondrial ATP. If the transported adenine nucleotides equilibrate with the intramitochondrial pool, [14C]ADP0 would be expected to appear with a lag phase that corresponds with the time needed for the radioactive labelling of the intramitochondrial adenine nucleotide pool. 2. The rates of formation of [14C]ADP outside the mitochondria after addition of [14C]ATP during the steady-state uncoupler-induced ATP hydrolysis catalysed by rat-liver mitochondria at 0 degree C were measured. 3. In the presence of carbonyl cyanide m-chlorophenylhydrazone the time course of the [14]ADPo formation was the same as that predicted on the basis of the above assumption. 4. In the presence of the less effective uncoupler, 2,4-dinitrophenol, the time course of [14C]ADPo formation was not consistent with the theoretical predictions: no lag phase was present and the measured rate was higher than the maximal calculated rate. These results can be explained by assuming a functional interaction between the adenine nucleotide translocator and the mitochondrial ATPase (F1). 5. It is concluded that under phosphorylating as well as dephosphorylating conditions, the adenine nucleotide translocator and the mitochondrial ATPase can be functionally linked to catalyse phosphorylation or dephosphorylation of extramitochondrial ADP or ATP, without participation of the intramitochondrial adenine nucleotides.

Adenosine Diphosphate↗
PubMed1976

Malplacement of endocardial pacemaker electrodes in the middle cardiac vein.

Endocardial transvenous pacemaker electrodes have been implanted in 246 patients owing to symptomatic bradycardia. The electrode was malplaced in the middle cardiac vein in 12 patients. All the patients were on stable ventricular pacing at the time of implantation. The electrodes were still in function in six patients after 5-96 months (average 55). The electrode was replaced in three patients owing to the occurrence of exit block. Three patients died during the period under study. Right-sided bundle branch block (RBBB) configuration of paced QRS complexes was observed on the ECG when the electrode was malplaced in the middle cardiac vein. As RBBB only occurs with malplacement, the ECG can be employed to ensure correct positioning of the electrode.

Aged↗