Penicillamine administration reverses the inhibitory effect of hyperhomocysteinaemia on endothelium-dependent relaxation and superoxide formation in the aorta of the rabbit

Although hyperhomocysteinaemia is a risk factor for cardiovascular disease, the mechanisms underlying this association have not been elucidated. It has been demonstrated, however, that copper augments the inhibitory effect of homocysteine on nitric oxide (NO)-mediated relaxation of the rat aorta through increased superoxide formation, which reacts with NO thereby reducing the bioavailability of NO. Since it follows that the administration of a copper chelator may blunt the pathogenic impact of hyperhomocysteinaemia, in vivo, the effect of penicillamine administration on NO-dependent relaxation and superoxide formation in the aortae of hyperhomocysteinaemic rabbits was studied. New Zealand White rabbits were fed a methionine-rich (20 g/kg chow) diet for 1 month+/-penicillamine administered orally (10 mg/kg/day) and aortic relaxation elicited with acetylcholine and superoxide measured. The role of NADPH oxidase was also studied using a range of inhibitors and western analysis of gp47(phox) (a catalytic subunit of NADPH oxidase). The methionine-rich diet markedly increased plasma total homocysteine levels. In hyperhomocysteinaemic rabbits there was a marked reduction of acetylcholine-stimulated relaxation and an increase in superoxide formation that were both inhibited with superoxide dismutase and apocynin, an NADPH oxidase inhibitor. Gp47(phox) expression was also increased in aortae from methionine fed rabbits. Penicillamine administration significantly reduced plasma total copper in methionine-fed rabbits compared to controls. Impaired acetylcholine-stimulated relaxation, increased superoxide formation and increased gp47(phox) expression in aortae from methionine-fed rabbits was reversed by penicillamine administration. These data indicate that hyperhomocysteinaemia augments the formation of arterial superoxide through an increase in NADPH oxidase expression/activity which in turn reduces NO bioavailability. Since these effects were reversed by penicillamine, these data consolidate the hypothesis that copper plays a role in mediating homocysteine-induced vasculopathy.     

Corticosteroid determination in skin preparations by a reaction rate method

A reaction rate method for the determination of beta-methasone, betamethasone valerate, triamcinolone acetonide, and fluocinolone acetonide is described. The method is based on a modification of the widely accepted blue tetrazolium reaction. Analysis times of 30--70 sec are required. Relative standard deviations of 0.3--1.9% are obtained, and the analytical working curves are linear. Analysis of pharmaceutical skin preparations by the new method gave results that correlated well with the time-consuming standard equilibrium method. Analysis of betamethasone and betamethasone valerate mixtures by measuring absorbance values at two different times was performed also.     

Application of automated flow injection analysis (FIA) to dissolution studies

The application of FIA to dissolution studies is described. Propantheline bromide, salicylamide and sulfamethizole were chosen as model drugs to investigate the utility of FIA method for dissolution studies. In each case the FIA system with the appropriate chemistry manifold was coupled with the rotating basket apparatus, A fully automated monitoring of dissolution rates was achieved. A complete dissolution profile in tabulated form is provided by the computer of the system at the end of the experiment.

Automation of any type of solid dosage forms agitation technique can be easily acquired by adapting a FIA system.     

Automated flow-injection spectrophotometric determination of catecholamines (epinephrine and isoproterenol) in pharmaceutical formulations based on ferrous complex formation

A novel automated flow-injection spectrophotometric method for the determination of catecholamines (epinephrine and isoproterenol) has been developed based on the formation of their coloured complexes with Fe(II) in aminoacetic-carbonate buffer pH 8.3 and measuring of the absorbance peaks at the lambda(max) of 530 nm. A fully automated FIA system controlled by home-made software (FIA-MOD) was used for optimising the chemical and manifold parameters and running of routine measurements. The calibration graph was linear in the range of 5-200 mg l(-1) for epinephrine with an RSD of 0.24% (n = 5; c = 150 mg l(-1)) and 10-300 mg(-1) for isoproterenol with an RSD of 0.13% (n = 5; c = 200 mg l(-1)). Measurement throughput was 120 h(-1) ensuring a sample throughput of 40 h(-1) analysed in triplicate. Common excipients for tablets and injections were found not interfering. The proposed method was applied for the assay of various commercial pharmaceutical formulations containing epinephrine and isoproterenol and for the content uniformity test for the isoproterenol tablets. The assay results with RSD 2-4% (n = 3) were comparable with those obtained with the official USP XXIII methods (mean difference 1.9%).     

Contemporary accuracy of death certificates for coding prostate cancer as a cause of death: Is reliance on death certification good enough? A comparison with blinded review by an independent cause of death evaluation committee

Background:

Accurate cause of death assignment is crucial for prostate cancer epidemiology and trials reporting prostate cancer-specific mortality outcomes.

Methods:

We compared death certificate information with independent cause of death evaluation by an expert committee within a prostate cancer trial (2002–2015).

Results:

Of 1236 deaths assessed, expert committee evaluation attributed 523 (42%) to prostate cancer, agreeing with death certificate cause of death in 1134 cases (92%, 95% CI: 90%, 93%). The sensitivity of death certificates in identifying prostate cancer deaths as classified by the committee was 91% (95% CI: 89%, 94%); specificity was 92% (95% CI: 90%, 94%). Sensitivity and specificity were lower where death occurred within 1 year of diagnosis, and where there was another primary cancer diagnosis.

Conclusions:

UK death certificates accurately identify cause of death in men with prostate cancer, supporting their use in routine statistics. Possible differential misattribution by trial arm supports independent evaluation in randomised trials.     

Evaporative light scattering detection (ELSD): a tool for improved quality control of drug substances

Thanks to the recent technological advancements, evaporative light-scattering detection (ELSD) is regarded as a valuable alternative to UV detection for liquid chromatographic analysis of substances that do not contain a chromophore. In the field of substances for pharmaceutical use, LC-ELSD appears to be suitable for aminoglycosides, most of which (for ex. gentamicin) are presently controlled in the Ph. Eur. by pulsed amperometric detection. Other substances (ex sugars, triglycerides) presently employing refractometrric detection, could be conveniently analysed by LC-ELSD. ELS detection is regarded as robust and relatively simple, although not particularly sensitive. A key feature of ELSD is that - unlike refractometry - it can operate in gradient mode, thus allowing application of more selective liquid chromatographic methods. ELSD can also be used to set up MS-compatible methods, as the mobile phase constraints are essentially the same. Due to all the above, ELSD is becoming increasingly used in pharmacopoeial methods.