This simple solid-phase chemiluminescence immunoassay for measurement of progesterone in extracts of venous plasma has sensitivity and precision similar to that of conventional radioimmunoassay with use of a tritiated antigen. The labeled antigen, 11 alpha-progesteryl-2-succinoyltyramine-4-(10-methyl)-acridini um-9-carboxylate, and a monoclonal antibody to progesterone-11 alpha-succinyl-bovine serum albumin are incubated with a 100-microL aliquot of plasma extract (equivalent to 20 microL of plasma) and 50 microL of a suspension of an IgG fraction of a donkey antiserum to mouse immunoglobulins, covalently attached to cellulose particles. After the antibody-binding reaction (60 min at 4 degrees C), 1 mL of phosphate buffer is added to each tube, the tubes are centrifuged (5 min, 1500 X g), and the supernatant fluid is aspirated. The washing step is repeated and diluted hydrochloric acid (50 mmol/L, 50 microL) is added to the pellet. Luminescence is initiated by oxidation with dilute sodium hydroxide/hydrogen peroxide. The signal is integrated over 10 s. The light yield is inversely proportional to the progesterone concentration in the standard or sample.
An extracellular dextranase (EC 3.2.1.11) was purified approximately 75-fold from cell-free culture filtrates of Fusarium moniliforme. The purified dextranase was of the endo type, and isomaltose was identified as the primary end product of dextran hydrolysis. The molecular weight of the dextranase was determined to be 39,000 by gel permeation chromatography. The enzyme was most active at pH 5.5, and the temperature optimum was near 55 C. Activity was not inhibited by either ethylenediaminetetraacetic acid or iodoacetate. The Km for dextran with an average molecular weight of 10,000 was estimated to be 1.1 X 10-4 M. The electrophoretic mobility of the dextranase was distinctly different from that of a Penicillium-derived commercial dextranase. The F. moniliforme dextranase was also found to differ from the commercial preparation by its greater relative activity against glucans isolated from Streptococcus mutans.
BackgroundWhile type 1 diabetics often require self-monitoring of blood glucose (SMBG), the evidence for tight blood glucose monitoring in non-insulin treated type 2 diabetes mellitus (T2DM) patients is limited. In addition to its lack of cost-effectiveness, unnecessary blood glucose monitoring may also result in anxiety and decreased quality of life. In this retrospective audit, we assessed SMBG prescribing practice at one general practice against guidelines from the National Institute for Health and Care Excellence (NICE).
MethodsA systematic search of T2DM patients diagnosed at a general practice in London, United Kingdom, in the last 10 years was undertaken. A total of 146 patients fulfilled these criteria, of which 100 patients were randomly selected for inclusion in this audit. Medical notes were reviewed and collated for analysis.
ResultsOnly 85% of patients with T2DM were being managed in accordance with the NICE guidelines on SMBG, while 15% were not. It was more common for patients who did not need monitoring to be inappropriately prescribed SMBG (10%) than it was for patients who needed monitoring to be under-prescribed SMBG (5%). The reasons for prescribing SMBG were often left undocumented.
ConclusionAdherence to the NICE guidelines is subpar. Recommended solutions include educating healthcare professionals involved in the prescribing of SMBGs, regular reviews of the continued necessity of SMBG, and digital alerts on e-prescribing systems.
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