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V Marks

Publications and source records attributed to V Marks.

At least 181 records · Page 10Linked to original sources

Determination of caffeine, theophylline and theobromine in serum and saliva using high-performance liquid chromatography.

A method is described for the measurement of theobromine, theophylline and caffeine in serum and saliva by high-performance liquid chromatography (HPLC). A chloroform/isopropanol extract (85:15, v/v) is evaporated to dryness and chromatographed on a 100 X 4.5 mm id Hypersil octadecylsilane column with UV detection at 280 nm. Theobromine, theophylline, caffeine and the internal standard proxyphylline are satisfactorily resolved with an elution system of acetonitrile/tetrahydrofuran/50 mM acetate buffer, pH 4.0, (4:1:95, v/v). No interference is observed from the presence of xanthine metabolites or any of a number of common drugs examined. A good correlation was observed between the concentrations of caffeine in serum and in saliva suggesting that salivary measurements may be useful for the study of caffeine pharmacokinetics in man. Caffeine levels determined by the HPLC procedure described here agreed well with those obtained by a radioimmunoassay method. The method is also suitable for determining the xanthine content of beverages by direct injection of diluted samples.

Caffeine↗

Measurement of plasma caffeine concentrations by substrate labelled fluoroimmunoassay.

Abuse or misuse of caffeine has been implicated in the pathogenesis of a variety of clinical disorders and measurement of its plasma concentration is likely to become important in clinical laboratories in the future. A rapid, sensitive and easily performed method for the measurement of caffeine in biological fluids by the technique of fluoroimmunoassay is described. The results produced are precise and agree well with those obtained by radioimmunoassay or HPLC. Cross-reactivity of the antiserum with other substituted xanthines and uric acids is negligible for the major metabolites of caffeine but there is significant cross-reactivity with theophylline at concentrations likely to be found when this drug is being administered therapeutically. The measurement of caffeine by fluoroimmunoassay has a place in the clinical laboratory particularly where small batches of samples are to be analysed.

Binding Sites, Antibody↗

Plasma immunoreactive gastric inhibitory polypeptide in obese hyperglycaemic (ob/ob) mice.

Gastric inhibitory polypeptide (GIP), a recognized component of the enteroinsular axis, is raised in the plasma and intestine of obese hyperglycaemic (ob/ob) mice. To evaluate the control of plasma GIP and its role in the hyperinsulinaemia of the ob/ob syndrome, GIP and insulin were determined at different ages in fed mice, and at 10-12 weeks of age after fasting/refeeding and administration of GIP, different nutrients and insulin to mice fasted for 18 h. Plasma GIP and insulin were raised in adult (10- and 20-week-old) compared with younger (3- and 5-week-old) mice, although GIP was not increased in the presence of hyperinsulinaemia at 3 weeks of age. Fasting suppressed and refeeding promptly restored plasma GIP and insulin concentrations. Administration of GIP to mimic postprandial concentrations evoked a marked but transient insulin response which was protracted in the presence of rising hyperglycaemia. Orally administered fat, glucose and amino acids raised GIP concentrations with fat having a particularly strong effect. Glucose and amino acids also evoked prominent increases of insulin, but fat produced only a small rise in insulin in the absence of increasing glucose concentrations. Consistent with glucose-potentiation, a mixture of all three nutrients greatly augmented the insulin response without further increase of plasma GIP. Glucose-induced increase in endogenous insulin and doses of exogenous insulin up to 100 units/kg did not suppress basal, fat-stimulated or glucose-stimulated GIP release. The results indicate that raised GIP concentrations make an important contribution to the hyperinsulinaemia and related metabolic abnormalities of the ob/ob syndrome.

Amino Acids↗

Effects of physical training on insulin, connecting peptide (C-peptide), gastric inhibitory polypeptide (GIP) and pancreatic polypeptide (PP) levels in obese subjects.

Ten severely obese women were subjected to physical training for three months on ad libitum diet. Under metabolic ward conditions oral glucose tolerance test was performed before and after the training period with the same energy intake quantitatively and qualitatively, and glucose, insulin, connecting (C)-peptide, gastric inhibitory polypeptide (GIP) and pancreatic polypeptide (PP) were determined. In confirmation of previous work, physical training caused no decrease in body fat in these severely obese subjects, and no change in body cell mass or glucose tolerance, while insulin and blood pressure decreased. The control of dietary conditions demonstrated that the latter phenomena were not due to quantitative or qualitative changes in the diet. C-peptide concentrations decreased also, indicating effects of physical training in obesity on insulin production. GIP is believed to be a gastrointestinal factor facilitating insulin secretion (Incretin). Previous work has indicated that gastrointestinal factor(s) are involved in the insulin lowering effect seen after physical training. It is possible that GIP is contributing to this phenomenon.

Adipose Tissue↗

Effect of pretreatment with a high fat diet on the gastric inhibitory polypeptide and insulin responses to oral triolein and glucose in rats.

Male Wistar rats were pretreated with 3 ml triolein orally for 4 days in addition to their normal diet. A similar control group were allowed free access to normal laboratory food. When given an oral fat load (1 ml triolein) plasma gastric inhibitory polypeptide (GIP) and triglyceride levels were significantly higher in the fat pretreated group. Inhibition of fat-stimulated GIP release by exogenous insulin was demonstrated in the untreated control group (plasma GIP: 663 +/- 49 versus 853 +/- 92 ng/l, mean +/- SEM p less than 0.025), but pretreatment with an oral fat load abolished this effect (plasma GIP: 1008 +/- 95 versus 1116 +/- 100 ng/l, p NS). Plasma glucose levels were significantly higher in fat pretreated rats given oral fat and intraperitoneal insulin compared with untreated controls (plasma glucose nadir 2.6 +/- 0.48 versus 1.6 +/- 0.15 mmol/l, p less than 0.05). Fat-pretreated rats showed significantly higher insulin and glucose levels compared with the untreated rats when given oral glucose (plasma insulin: 6.2 +/- 1.2 versus 2.5 +/- 0.59 micrograms/l, p less than 0.01; plasma glucose: 10.2 +/- 0.39 versus 8.9 +/- 0.41 mmol/l, p less than 0.025). Pretreatment of rats on a high fat diet causes (1) increased GIP secretion in response to an oral fat load, (2) abolition of the feed-back inhibition of exogenous insulin on fat-stimulated GIP release, and (3) some degree of insulin resistance.

Animals↗

Plasma levels and response to prednisolone therapy in rheumatoid arthritis.

Plasma prednisolone levels were measured in parallel with clinical and laboratory assessments in seven patients with rheumatoid arthritis on single daily doses of prednisolone which were reduced at weekly intervals from 10 mg. to 7, 6 and 5 mg. A significant negative rank correlation was observed between prednisolone dose and the duration of morning stiffness (p less than 0.05). No significant correlation was observed between plasma prednisolone levels, other clinical assessment criteria or laboratory indices.

Adult↗

Diurnal variation in prednisolone kinetics.

Plasma concentrations of free and total prednisolone were measured after oral doses at four time points to investigate the possibility of a diurnal variation in the drug's kinetics. There were marked differences in plasma prednisolone concentrations, clearance rates, and bioavailability of both free and total fractions at different times of the day. Changes in the protein binding characteristics of prednisolone with clock time resulted in marked differences between the kinetics of free and total prednisolone. It is recommended that for maximum efficacy and minimum toxicity prednisolone therapy be confined to once-daily dosing in the morning.

Adult↗

Measurement of serum theophylline concentrations using a modified Ames TDA system.

Results are presented which show that the Ames TDA Theophylline kit provides an accurate and reliable method of measuring the concentration of theophylline in plasma. It may be used with a shortened incubation time (5 min) without loss of accuracy, and a single calibration curve can be used over a 3-week period. There is an acceptable precision at all assayed levels (coefficient of variation, 3.3%) and results obtained for patients' samples correlate well with those obtained using liquid chromatography (y = 0.02 + 1.06x, r = 0.99) and enzyme immunoassay (y = 0.58 + 1.03x, r = 0.99).

Bilirubin↗

Plasma prednisolone studies in rheumatic patients.

Prednisolone concentration in plasma after a daily maintenance steroid dose was monitored in 83 patients with rheumatic diseases. Although no restrictions were imposed on the intake of food or drink, plasma drug levels tended to peak at 1 h after the ingestion of the standard tablets, whereas intestinal absorption of the enteric-coated preparation was found to be most unpredictable. A profound individual variation was observed in plasma prednisolone concentrations, and neither total nor unbound drug levels showed any consistent relationship with the size of the prednisolone dose or the control of the disease activity. The basal cortisol production was suppressed in one-third of the patients. There is no support from this study for the concept of an 'optimum therapeutic range' of plasma prednisolone for the treatment of rheumatic disease.

Adult↗

Clinical pathology of alcohol.

There is good though not conclusive evidence that a small to modest average daily intake of alcohol--that is, 20-30 g/day is associated with increased longevity due mainly to a reduction in death from cardiovascular disease. Larger average daily alcohol intakes--especially those in excess of 60 g/day for men and 40 g/day for women--are associated with gradually increasing morbidity and mortality rates from a variety of diseases. Alcohol may be unrecognised as the cause of somatic disease, which can occur without overt psychosocial evidence of alcohol abuse, unless the index of suspicion is high and a thorough drink history obtained. Laboratory tests for the detection and/or confirmation of alcohol abuse are useful but subject to serious limitations being neither as sensitive nor specific as sometimes believed. The value of random blood and/or breath alcohol measurements, in outpatients, as an aid to diagnosis of alcohol-induced organic disease is probably not sufficiently appreciated and, though relatively insensitive, is highly specific.

Acetaldehyde↗

A critical evaluation of a new fluorescence immunoassay system for the measurement of serum phenytoin concentrations.

Results are presented which show that the Ames TDA tm phenytoin kit may be used with a shortened incubation time (5 minutes) and that, using a Fluorostat tm instrument, a single calibration curve can be stored and used effectively over a three-week period. The importance of temperature control when using a single calibration curve is emphasised. An unexplained finding was that higher results were obtained for phenytoin concentrations up to about 15 mg/l with increasing incubation temperature. There was acceptable precision of measurement at all levels of phenytoin concentration, and there was no significant interference by haemoglobin, bilirubin, or lipids. The TDA results correlated well with those obtained using liquid chromatography (y = 0.39 + 0.98x, r = 0.98, where y = TDA and x = HPLC) and enzyme immunoassay (y = 0.08 + 0.93x, r = 0.96, where y = TDA and x = EIA).

Bilirubin↗

Abnormalities of GIP in spontaneous syndromes of obesity and diabetes in mice.

The role of GIP in the pathogenesis of spontaneous syndromes of obesity-diabetes was examined in ob/ob mice of the Aston stock and db/db mice of the C57BL/KsJ background. Compared with lean controls, fed adult ob/ob and db/db mice, respectively, exhibited 1.8-fold and 2.1-fold increases in body weight, 1.8-fold and 2.8-fold elevations of plasma glucose, and 15.4-fold and 5.6-fold elevations of plasma insulin. As indicated by the relative magnitude of the hyperglycemia and hyperinsulinemia, db/db mice displayed a particularly severe form of diabetes. Plasma GIP concentrations of ob/ob and db/db mice were elevated 15.1-fold and 6.2-fold, respectively; the increments closely corresponded with the degrees of hyperinsulinemia. Small intestinal weight was increased 1.4-fold and 1.8-fold in ob/ob and db/db mice, respectively, but the intestinal GIP content expressed as microgram/g intestine or microgram/intestine was raised only in ob/ob mice (1.9-fold and 2.8-fold, respectively). Since glucose stimulation of insulin release is defective in both mutant strains, the results strongly implicate pathologically raised GIP concentrations in the hyperinsulinemia and related metabolic abnormalities of the obesity-diabetes syndromes. It is suggested that hypersecretion of GIP results in part from loss of normal feedback inhibition by endogenous insulin.

Animals↗