Effects of EDTA on chemiluminescent immunoassay measurement of ACTH, cortisol, and thyroid hormones in dogs.

When measuring blood hormones, pre-analytical sample handling can impact the quality of the results. Previous studies have shown improved stability of canine cortisol in ethylenediaminetetraacetic acid (EDTA) plasma compared to serum and interchangeability of serum and plasma when cortisol is measured by radioimmunoassay. Additionally, cortisol samples were also interchangeable when measured by chemiluminescent immunoassay if the EDTA concentration was consistent with that of optimally filled tubes, whereas excess EDTA interfered with the measurement of cortisol and serum and EDTA plasma were not interchangeable when measuring total thyroxine (TT4). The main limitation of these studies was that they were performed by spiking pooled serum samples with EDTA or in previously collected blood samples submitted to a clinical pathology laboratory. The purpose of the present study was to evaluate the effect of EDTA on the measurement of adrenocorticotropic hormone (ACTH), cortisol, TT4, free thyroxine (FT4), and thyroid stimulating hormone (TSH) in healthy dogs using the Siemens IMMULITE 1000. Whole blood from forty dogs was aliquoted into three Monoject sample tubes: no additive, completely filled EDTA tube, and 50% filled EDTA tube. Handling and storage conditions were identical, and all samples were analyzed on the same day. Bland-Altman plots and Passing-Bablok regression were used to assess agreement and risks for error, respectively. Proportional errors were found between serum and plasma samples for ACTH, cortisol, TT4, FT4, and TSH; systematic errors were also found for FT4. There was poor agreement and clinically significant differences between the measured concentrations of all hormones in serum and plasma, proving that these sample types are not interchangeable. Incompletely filled EDTA tubes were associated with significantly lower ACTH concentrations compared to completely filled EDTA tubes. When measured by chemiluminescent immunoassays that utilize alkaline phosphatase at the reporter enzyme, serum should be used for cortisol, TT4, FT4, and TSH, while plasma from completely filled EDTA tubes should be used for ACTH.

[Measuring visual field loss by automated perimetry in chronic glaucoma stabilized by trabeculectomy]. / Evolution du champ visuel en périmétrie automatisée dans le glaucome chronique équilibré par trabéculectomie.

The progression of field loss was determined by Humphrey Automated threshold perimetry in the short and intermediate term after surgical trabeculectomy for chronic glaucoma: mean indices (MD, PSD et FOV) were studied before and after normalisation of the intraocular pressure. Risk factors such as secondary glaucoma, previous visual field defects and cardiovascular risk factors were significantly correlated to poor or bad progression of field loss: the mean variation of MD = +1.23 dB +/- 2.84 (p < 1.5%), the mean variation of PSD = 0.50 dB +/- 1.37 (p < 6%), but the mean variation of FOV was not significant; so 36% of field defects were improved, 42% were stabilized and 22% were aggravated. When the risk factors were absent, 86% of field losses were stabilised but when cardiovascular factors were present, only 64% were stabilised. These risk factors can also be used to ascertain the prognosis of post-surgical progression of visual field, as already shown in previous reports. Automated threshold perimetry was useful to study the visual field progression with standardized and reliable numerical parameters, allowing statistical computerised management of the patients with glaucoma.

Folliculitis ulerythematosus reticulata: a report of four cases and brief review of the literature.

Folliculitis ulerythematosus reticulata (FUR) is an uncommon genodermatosis best classified as one of the keratosis pilaris atrophicans group of disorders. It is characterized by erythema and follicular plugging of the cheeks that evolves into honeycomb atrophy. Associated cutaneous and visceral abnormalities can occur. This report describes four patients with FUR, one with an unaffected twin, a finding not previously reported. The significance of an early and accurate diagnosis is emphasized.

Comparison of high pressure liquid chromatographic and chemical methods for vitamin D3 concentrates. II. Collaborative study.

A collaborative study was carried out which compared the official chemical method, 43.B14-43.B24, the official rat bioassay, 43.165, and the high pressure liquid chromatographic method for vitamin D3 resin, vitamin D3 resin in oil, and dry concentrate. A total of 340 samples were distributed to 17 collaborators for analysis. Five laboratories performed both the chemical and HPLC methods on 5 sets of blind duplicates. A 2-way analysis of variance comparing both methods for each sample showed a significant (P less than 0.01) difference between methods only for Sample 5. When the 2 methods were compared over all the samples, no significant (P less than 0.05) difference was found. Except for Sample 5, there were no differences in the repeatability of the methods. Per cent recoveries on Sample 3, which contained exactly 0.200 X 10(6) IU/g, showed 98.2% for the chemical method and 100.6% for the HPLC method for the 5 laboratories that performed both methods. The assay results of the HPLC and chemical methods are in good agreement with those found by the biological assay on Samples 1-4, but not for Sample 5. Evidence indicates that Sample 5 degraded partially to isotachysterol, and while the HPLC method yielded a reasonable value on this material, the chemical method erroneously showed full potency. An amendment is included for the collaboratively studied HPLC method which detects and eliminates 5,6-trans-vitamin D3, a possible interferant.