Analytical and clinical comparison of different immunoassay systems for the detection of antiphospholipid antibodies.

INTRODUCTION: We evaluated analytical and clinical performances of IgG and IgM anticardiolipin (aCL) antibodies and anti-ß2-glycoprotein I (a-ß2GpI) antibodies and upper limit reference ranges (99th percentiles) in comparison with manufacturer's cutoff values with different commercial methods. METHODS: We assayed aCL and a-ß2GpI in serum samples from 30 healthy individuals, 77 patients with antiphospholipid syndrome (APS) diagnosed according to the Sydney criteria, 51 patients with autoimmune diseases, eight patients with previous thrombotic events, six patients with other diseases, and 18 patients with infectious diseases, using ELISA Inova Diagnostics; EliA Phadia Laboratory Systems; CliA Zenit-RA; and CliA Bio-Flash. RESULTS: Anticardiolipin and a-ß2GpI IgG and IgM immunoassays showed good analytic performances with both 99th percentile and manufacturer's cutoff reference values. Our results showed fair to moderate agreement among assays. In-house cutoff values gave significantly better performances only for a-ß2GpI IgG with all the immunoassays analyzed with the exception of Inova CliA Bio-Flash where we obtained the same performances with in-house and manufacturer's cutoffs. CONCLUSIONS: By guidelines, all laboratories are strongly advised to validate/verify the manufacturer's cutoff values. We recommend establishing low-positive, medium-/high-positive, and high-positive CliA IgG aCL and a-ß2GpI ranges in order to help clinicians in the diagnosis and treatment of APS.

Impact of perioperative symbiotic therapy on infectious morbidity after Hpb Surgery in jaundiced patients: a randomized controlled trial.

This study aimed at evaluating whether the administration of symbiotic therapy in jaundiced patients could reduce their postoperative infectious complications. The study was conducted between November 2008 and February 2011. Jaundiced patients scheduled for elective extrahepatic bile duct resection without liver cirrhosis, intestinal malabsorption or intolerance to symbiotic therapy were randomly assigned to receive [Group A] or not [Group B] symbiotics perioperatively. The primary endpoint was the infectious morbidity rate. Forty patients were included in the analysis (20 in each group). The patients in Group B presented a higher overall morbidity (70 vs 50%) and infectious morbidity rate (50 vs 25%), but the differences were not significant. Eleven patients in Group A (Group ndA) and 13 in Group B (Group ndB) did not receive preoperative biliary drainage. The results of the two groups were comparable. Infectious complications were higher in Group B [5 (34%) vs 0, p = 0.030], while the prevalence of natural killer (NK) cells was higher in Group ndA the day before surgery (17% ± 5.1 vs 10% ± 5.3, p < 0.01) and on post-operative day (POD) 7 (13.1% ± 4.1 vs 7.7% ± 3.4, p < 0.01). The rates of lymph node colonization were similar. The symbiotic therapy failed to reduce the rate of infectious morbidity in jaundiced patients. Further studies investigating the place of symbiotic in no-drainage patients are required.

Application of the use of high-throughput technologies to the determination of protein structures of bacterial and viral pathogens.

The Structural Proteomics In Europe (SPINE) programme is aimed at the development and implementation of high-throughput technologies for the efficient structure determination of proteins of biomedical importance, such as those of bacterial and viral pathogens linked to human health. Despite the challenging nature of some of these targets, 175 novel pathogen protein structures (approximately 220 including complexes) have been determined to date. Here the impact of several technologies on the structural determination of proteins from human pathogens is illustrated with selected examples, including the parallel expression of multiple constructs, the use of standardized refolding protocols and optimized crystallization screens.

Spot urinary iodine concentration as a measure of dietary iodine, evaluated in over 3800 young male subjects undergoing medical check-up preliminary to military enrolment in Piemonte and Aosta Valley (Italy).

The aim of the study was to assess iodine status in over 3800 young male subjects aged 18, living in 6 different provinces of Piemonte and in Aosta Valley, Italy. A cross-sectional study on 3837 young male subjects undergoing medical evaluation preliminary to military enrolment was performed. Spot urine samples were obtained in the morning hours and urinary iodine was measured by a colorimetric method. As outcome measure iodine status, based on spot urinary iodine median concentration, categorised as sufficient (>99 microg/l), mild deficiency (50-99 microg/l), moderate deficiency (20-49 microg/l) and severe deficiency (<20 microg/l), was obtained. Median urinary iodine concentration was 101.8 microg/l in our sample. Moderate-to-severe iodine deficiency was found in <10% of all subjects. Mild iodine deficiency was found in <40% of the subjects, with greater variability within the provinces considered. For each geographical area significant differences can be observed in more than 50% of the comparisons of the confidence intervals related to the frequencies of samples with spot urinary iodine concentration below 100 microg/l. The high frequency of mild iodine deficiency found in our sample suggests that dietary sources of iodine in Piemonte, Italy, have improved since the last evaluation 25 yr ago (2) but may still be insufficient. Further population studies are required.

[Intact whole bioactive parathormone: problems arising from comparing different methods]. / Paratormone intatto intero bioattivo: le problematiche emergenti dal confronto.

BACKGROUND: Parathyroid hormone (PTH) has important applications in the nephrological clinical practice. Because assays of Intact PTH (I-PTH) are liable to interferences by N-truncated fragments, a novel method for whole-(1-84) PTH has been proposed. This study is aimed at comparing the latter with some of the previous I-PTH assays. For each method the results are referred to pertinent markers of mineral metabolism. METHODS: We enrolled 171 subjects, including 56 healthy controls (C), 65 calcium stone- formers (CaSF), 40 haemodialysis patients (HD), 10 with primary hyperparathyroidism (PHP). On blood samples we measured: I-PTH by four methods (N-Tact, Advantage, Elecsys, Scantibodies), whole-(1-84) PTH, defined as CAP (Cyclase Activating PTH), total and ionised calcium, phosphate, vitamin D, osteocalcin and Crosslaps. The difference between I-PTH and CAP Scantibodies is defined as CIP (Cyclase Inhibiting PTH). RESULTS: Despite relating to each other (r>0.97) PTH values varied remarkably among methods. For all methods, the reference intervals differed from those provided by the producer. Assuming these new ranges, 10 CaSF had over-range values not always associated with abnormalities of mineral metabolism. One of the PHP patients was normal for I-PTH with 2/4 methods. In HD the differences among methods were even greater, there were inverse (p<0.05) and direct (p<0.001) relationships with ionised calcium and osteocalcin-crosslaps, respectively. The CAP/CIP ratio was lower in low bone turnover patients, but the two subgroups widely overlapped. CONCLUSIONS: This study indicates that the reliability of I-PTH assays is still unsatisfactory, and none of the four methods emerged as the best. Assay for CAP only improves diagnostic efficiency, whereas the CAP/CIP ratio does not exhibit powerful discriminating capacity. Our suggestion is that each Centre should establish its own reference ranges. PTH assay should always be coupled with measurements of other markers of mineral metabolism as well as renal function.

Dehydroepiandrosterone concentration in breast cancer tissue is related to its plasma gradient across the mammary gland.

Dehydroepiandrosterone (DHEA) has been shown to affect the growth of mammary carcinomas both in vitro and in vivo. In humans, very high levels of DHEA and/or dehydroepiandrosterone sulfate (DHEAS) have been found in breast tissues and secretions, and epidemiological studies suggest a role of these steroids in the modulation of breast cancer growth. An uptake from plasma and a transformation from precursors can be both postulated, but the main source of the adrenal C19 steroids found within the breast is debated. Attempting to clarify this point, in ten patients undergoing surgery for breast cancer we studied: a) DHEAS and DHEA concentrations in tumor tissue; b) the differences between DHEAS (or DHEA) concentration in peripheral venous plasma and that draining the affected breast, that we assume to reflect the arteriovenous gradient of these steroids; c) DHEA sulfatase activity in tumor tissue. Results show that DHEA sulfatase activity is not related to DHEAS or DHEA concentrations in breast cancer tissue. A negative DHEA plasma gradient across the breast is unveiled, whereas DHEAS levels are not different in blood supplying and draining the breast with cancer. The DHEA plasma gradient across the breast is positively related to DHEA concentration in tumor tissue. Data are consistent with the hypothesis that the plasma source contributes remarkably to DHEA found within breast cancer tissue.

Steroid gradients across the cancerous breast: an index of altered steroid metabolism in breast cancer?

The concentrations of 17 beta-estradiol, estrone, testosterone (T), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulphate, androstenedione (A), cortisol and prolactin (PRL) were determined in the peripheral venous blood and in the lateral thoracic vein of 14 premenopausal and 34 postmenopausal women who underwent surgery for a breast carcinoma. The difference between the two blood samples, defined as concentration gradient across the cancerous breast, was calculated for all hormones. A significant peripheral-local concentration gradient was found for DHEA and A both in pre- and postmenopausal patients, whereas for T it was observed only in postmenopausal subjects. Furthermore, DHEA and A gradients were correlated to the presence of estrogen receptors as determined by a radioligand binding assay. An inverse relationship between DHEA gradient and the expression of estrogen receptors was observed in premenopausal women, whereas in postmenopausal patients an opposite, although not significant, trend was found. These results suggest that in the cancerous breast: (1) DHEA, A and T (the latter only in postmenopause) could be taken up from plasma, and thus there could be a storage of these steroids inside the breast tissue and/or perhaps some alterations in their local metabolism; (2) androgens could play a different role in breast carcinogenesis in relation to the estrogen circulating levels and to the expression of estrogen receptors.

Iodine content of human tissues after administration of iodine containing drugs or contrast media.

Total iodine contents were determined in 209 bioptic or autoptic specimens of various extrathyroidal tissues. Fifty-two of the 80 subjects examined had no previous exposure to excessive iodine, 24 were tested after administration of x-ray contrast media and 4 after treatment with various iodine containing drugs. Unexposed subjects had tissue iodine contents (mean +/- SD) ranging from 0.85 +/- 0.17 microgram/100 g in the brain and 9.78 +/- 2.75 microgram/100 g in the liver of adults. A significantly lower iodine concentration was found in the liver of newborns (2.79 +/- 1.00 microgram, p less than 0.001). Most of the other tissues had iodine concentrations of 2-4 microgram/100 g. Subjects with previous exposure to iodine containing drugs or x-ray contrast media showed increased iodine contents of various degree in all examined tissues, including adipose tissue, bone, brain, kidney, liver, lung, skeletal muscle, skin and spleen. Accumulation of iodine in adipose tissue was still demonstrable more than two years after cholecystography.