Advances in immunoassay technology.

Major developments continue to be reported in key areas of immunoassay technology. Following the development of excellent signal generation methods, attention has shifted to the development of immunochemical methods and instrumentation to provide convenient systems of high performance. Important advances have been made in the design of immunochemical approaches that permit the replacement of competitive format assays for small molecules, such as drugs, metabolites and pollutants, with non-competitive formats, bringing advantages previously seen only with large molecular analytes. Bispecific antibodies and recombinant proteins are also beginning to impact immunodiagnostics, with the promise of even more highly specified reagents. Improvements in automation have brought the facility of homogeneous systems to high-throughput and high-performance heterogeneous systems. Similarly, 'point of need' testing continues to progress. Through all of these advances, systems are evolving according to the needs of users in terms of operator convenience, accuracy, specificity, speed, robustness, and sensitivity.

Eating disorder symptoms in former female college gymnasts: relations with body composition.

The purpose of this investigation was to describe eating disorder symptoms in 36.6 +/- 3.8-y-old former college gymnasts as well as relations between body dissatisfaction and body composition. Former college gymnasts (n = 22) and age-(mean +/- SE difference: 0.05 +/- 0.26 y), height-(0.47 +/- 0.75 cm), and weight-matched (2.20 +/- 0.30 kg) control subjects (n = 22) participated. Current and past symptoms were assessed by using the Eating Disorders Inventory-2 (EDI-2) and visual analog scales. EDI-2 body-dissatisfaction scores were correlated with assessments of body composition by dual-energy X-ray absorptiometry. Weight preoccupation was stable across the life span for control subjects but was lower before former gymnasts had begun gymnastics training and higher for former gymnasts when they were participating in college gymnastics (P = 0.03). Current levels of body dissatisfaction were more strongly related to actual minus ideal body-weight discrepancy scores (r = 0.77) than to percentage fat (r = 0.50) for the former gymnasts whereas the opposite was true for the control subjects (r = 0.51 and 0.77, respectively). These results suggest that symptoms of eating disorders abate after retirement from gymnastics and that concerns about achieving an ideal body may be a more important determinant of body dissatisfaction than percentage body fat for gymnasts.

Hyperprolactinemia. Long-term effects of bromocriptine.

Patients with hyperprolactinemia may be managed by pituitary surgery or irradiation, bromocriptine treatment, or a combination of these methods, and some patients remain untreated. Little is known of the long-term consequences of some of these therapeutic regimens. Forty-six hyperprolactinemic patients (40 female and six male) managed solely with bromocriptine or no treatment over a period of 12 months to six years were therefore evaluated in this study. Nine patients with radiologically normal pituitary fossae were untreated and 10 received bromocriptine, 7.5 to 10 mg daily, while 20 patients with radiologic evidence of a pituitary tumor were treated with bromocriptine, generally 10 to 20 mg daily. Patients were assessed clinically, biochemically, and radiologically before treatment and at least six weeks after discontinuation of therapy. A further seven patients were similarly assessed before and after eight bromocriptine-induced pregnancies. Symptoms persisted in the untreated group of nine patients, although menstruation returned in four of the females with previous amenorrhea; serum prolactin levels remained elevated, other pituitary function did not change, and pituitary fossae remained normal radiologically. In all patients treated with bromocriptine, symptoms improved irrespective of radiologic findings on the pituitary, and were abolished in 67 percent during treatment associated with a decrease in serum prolactin levels in all, and a return of levels to within normal limits in 80 percent of patients. Persistent side effects were usually dose-related, but remained troublesome in 13 percent. Bromocriptine-induced tumor regression was evident radiologically in all patients with suprasellar tumor tissue and in some with purely intrasellar adenomas. This effect occurred rapidly and persisted or increased throughout follow-up. On discontinuation of treatment, prolactin levels remained significantly lower than before therapy (mean 2,934 versus 5,052 mU/liter, p less than 0.05) but were within the normal range in only two patients. Other pituitary function was unaltered, or improved in some patients with definite tumors. Bromocriptine-induced pregnancy produced no permanent change in clinical, biochemical, or radiologic status. Long-term bromocriptine treatment for hyperprolactinemia is thus highly effective in alleviating symptoms and suppressing prolactin secretion, and induces persistent tumor regression on treatment without deterioration of other pituitary function in patients with macroadenomas. On discontinuation of therapy, however, hyperprolactinemia usually recurs, and treatment may therefore need to be continued for years.

Heparin resistance and decreased hepatic triglyceride hydrolase release during long-term estrogen-progestin treatment.

These studies were undertaken to define the mechanism for the depression of post-heparin triglyceride hydrolase activity in women treated with estrogen-progestin oral contraceptives. Six treated and six control women were studied. Total, protamine-inhibited, and protamine-resistant triglyceride hydrolase activities were measured after six different intravenous doses of heparin in each subject in order to determine the dose-response relationships for lipase release. As has been reported during short-term treatment with estrogens, long-term treatment with oral contraceptive agents is accompanied by selective depression of protamine-resistant (hepatic) lipase activity. This depression can be partly reversed by the administration of large heparin doses, but maximally releasing heparin does fail to restore postheparin protamine-resistant activity to control values. These data are compatible with the idea that the releasable pool of hepatic triglyceride hydrolase activity is diminished in women who receive oral contraceptive agents and that the pharmacokinetics of its release are altered in such a way that only relatively high concentrations of heparin displace the enzyme from this pool.

Seasonal variation of serum 25-hydroxyvitamin D in patients with chronic renal failure treated by regular haemodialysis.

Serum 25-hydroxyvitamin D was measured by a competitive protein-binding assay in 44 normal subjects, 60 uraemic patients on regular haemodialysis at different times of the year and in 13 non-dialyzed uraemic patients. The results obtained indicate that uraemic patients on regular haemodialysis have a mean serum 25-hydroxyvitamin D concentration comparable to controls and that they also exhibit a seasonal variation with a significant reduction during the winter months. However, serum 25-hydroxyvitamin D concentration remained essentially within the normal range and did not reflect the increased incidence of osteomalacia in these patients. In the 13 non-dialyzed uraemic patients, serum 25-hydroxyvitamin D concentrations were lower than in the dialyzed patients, but the explanation is not yet clear. This reduction in serum 25-hydroxyvitamin D was not accompanied by any osteomalacia. The results indicate that deficiency of 25-hydroxyvitamin D in our patients on regular haemodialysis is uncommon and clearly not the explanation of dialysis osteomalacia.

Serum adrenal androgens in women with primary operable breast cancer and their relationship with age and body mass index.

Several studies have found elevated levels of adrenal androgens in postmenopausal women and depressed levels in premenopausal women with breast cancer, suggesting a role for adrenal androgens in the aetiology of breast cancer. We have measured serum dehydroepiandrosterone sulphate and androstenedione in 81 women with primary operable breast cancer and 62 age-matched controls. Results showed that serum levels of both adrenal androgens fell significantly with age in women with breast cancer (P=0.003). However, no relationship was observed between serum adrenal androgen levels and body mass index in either women with breast cancer or controls. Dehydroepiandrosterone sulphate levels were elevated in postmenopausal women with breast cancer compared to controls, and this was not due to preoperative stress. No differences were observed in androstenedione levels between premenopausal or postmenopausal women with breast cancer and controls, nor were dehydroepiandrostenedione sulphate levels significantly different between premenopausal women with breast cancer and controls. These results suggest that dehydroepiandrosterone sulphate has a role in the aetiology of postmenopausal breast cancer.

Stimulation of cyclic AMP production in chick renal tissue by cadmium and manganese.

The effects of cadmium on production of cyclic AMP by partially purified chick renal plasma membrane preparations and binding of 125I-parathyrin to the membranes have been investigated. At certain concentrations Cd2+ ions (and Mn2+ ions) markedly stimulated the production of cyclic AMP by the tissue. It was found that concentrations of Cd2+ roughly in the same range were also capable of stimulating binding of 125I-parathyrin to the membrane preparations.

Moderate-intensity muscle pain can be produced and sustained during cycle ergometry.

PURPOSE: The aim of this experiment was to determine whether moderate-intensity quadriceps muscle pain could be produced and sustained during cycle ergometry. METHODS: 12 women volunteers completed a peak cycle ergometry test during which quadriceps muscle pain-intensity ratings were obtained using a 0-10 scale. On a subsequent day, participants cycled for 20 min beginning at the power output associated with quadriceps muscle pain threshold during the peak test. Participants manipulated power output during the first 10 s of each minute to produce and sustain moderate-intensity quadriceps muscle pain. During the last 20 s of each minute, VO2, ratings of quadriceps muscle pain intensity, and ratings of perceived exertion (Borg 6-20) were obtained. RESULTS: Moderate-intensity quadriceps pain ratings were achieved within 4 min, during which time the average power output was increased from 124 to 138 W. Pain-intensity ratings were unchanged from minutes 4 to 20 (F = 0.94; df = 16, 176; P = 0.52; eta2 = 0.079). Quadriceps RPE responses mimicked the pain responses. To sustain a moderate-intensity quadriceps pain participants reduced power output significantly (F = 7.75; df = 16, 176; P < 0.001; eta2 = 0.413) and in a linear fashion by 21.7 W (15.7%). The changes in % VO2peak were similar in pattern to the changes in power output but smaller in magnitude. CONCLUSIONS: 1) College women can produce and sustain a moderate-intensity quadriceps muscle pain during cycle ergometry, 2) moderate-intensity quadriceps muscle pain is sustained by reducing power output, and 3) sustaining moderate-intensity quadriceps muscle pain during short duration cycling is associated with a "hard" perceptual effort and 69-74% VO2peak.

Naturally occurring muscle pain during exercise: assessment and experimental evidence.

The objectives were: (i) to present a method for assessing muscle pain during exercise, (ii) to provide reliability and validity data in support of the measurement tool, (iii) to test whether leg muscle pain threshold during exercise was related to a commonly used measure of pain threshold pain during test, (iv) to examine the relationship between pain and exertion ratings, (v) to test whether leg muscle pain is related to performance, and (vi) to test whether a large dose of aspirin would delay leg muscle pain threshold and/or reduce pain ratings during exercise. In study 1, seven females and seven males completed three 1-min cycling bouts at three different randomly ordered power outputs. Pain was assessed using a 10-point pain scale. High intraclass correlations (R from 0.88 to 0.98) indicated that pain intensity could be rated reliably using the scale. In study 2, 11 college-aged males (age 21.3 +/- 1.3 yr) performed a ramped (24 W.min-1) maximal cycle ergometry test. A button was depressed when leg muscle pain threshold was reached. Pain threshold occurred near 50% of maximal capacity: 50.3 (+/- 12.9% Wmax), 48.6 (+/- 14.8% VO2max), and 55.8 (+/- 12.9% RPEmax). Pain intensity ratings obtained following pain threshold were positively accelerating function of the relative exercise intensity. Volitional exhaustion was associated with pain ratings of 8.2 (+/- 2.5), a value most closely associated with the verbal anchor "very strong pain." In study 3, participants completed the same maximal exercise test as in study 2 as well as leg cycling at 60 rpm for 8 s at four randomly ordered power outputs (100, 150, 200, and 250 W) on a separate day. Pain and RPE ratings were significantly lower during the 8-s bouts compared to those obtained at the same power outputs during the maximal cycle test. The results suggest that noxious metabolites of muscle contraction play a role in leg muscle pain during exercise. In study 4, moderately active male subjects (N = 19) completed two ramped maximal cycle ergometry tests. Subjects drank a water and Kool-Aid mixture, that either was or was not (placebo) combined with a 20 mg.kg-1 dose of powdered aspirin 60 min before exercise. Paired t-tests revealed no differences between conditions for the measures of exercise intensity at pain threshold [aspirin vs placebo mean (+/- SD)]: power output: 150 (+/- 60.3 W) versus 153.5 (+/- 64.8 W); VO2: 21.3 (+/- 8.6 mL.kg-1.min-1) versus 22.1 (+/- 10.0 mL.kg-1.min-1); and RPE: 10.9 (+/- 3.1) versus 11.4 (+/- 2.9). Repeated measures ANOVA revealed no significant condition main effect or condition by trial interaction for pain responses during recovery or during exercise at 60, 70, 80, 90, and 100% of each condition's peak power output. It is concluded that the perception of leg muscle pain intensity during cycle ergometry: (i) is reliably and validly measured using the developed 10-point pain scale, (ii) covaries as a function of objective exercise stimuli such as power output, (iii) is distinct from RPE, (iv) is unrelated to performance of the type employed here, and (v) is not altered by the ingestion of 20 mg.kg-1 acetylsalicylic acid 1 h prior to the exercise bout.

Pasteurella haemolytica as a co-pathogen in pullets and laying hens.

Pasteurella haemolytica was isolated from internal organs of laying hens and pullets of four flocks in which five incidents of increased mortality had occurred. There was inflammation of the trachea in all cases, and infectious laryngotracheitis virus was identified in three of the incidents. Rapid response to antimicrobial therapy was seen in two out of three outbreaks in the pullets. P. haemolytica was regarded as a significant secondary invader in these cases.

Vitamin D-binding protein gene microsatellite polymorphism influences BMD and risk of fractures in men.

UNLABELLED: Here we report the results of a vitamin D-binding protein gene microsatellite polymorphism study in 170 men, comprising healthy male subjects and men with osteoporosis-related symptomatic vertebral fractures. We confirm the results of an earlier study in a different cohort, showing relationship between certain genotypes of (TAAAn)-Alu repeats and reduced BMD and vertebral fractures. INTRODUCTION: Vitamin D-binding protein (DBP) plays a critical role in the transport and metabolism of metabolites of vitamin D, including the key calciotropic hormone 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3). METHODS: We have investigated intra-intronic variable tandem (TAAA)n-Alu repeat expansion in the DBP gene in 170 men, comprising healthy male subjects and men with idiopathic osteoporosis and low trauma fractures. RESULTS AND CONCLUSIONS: The predominant DBP-Alu genotype in the control subjects was 10/10 (frequency 0.421), whereas the frequency of this genotype in men with osteoporosis was 0.089. DBP-Alu alleles *10, *8 and *9, respectively, were the three commonest in both healthy subjects and men with osteoporosis. Allele *10 was associated with a lower risk of osteoporosis (OR 0.39, 95% CI 0.25-0.64; p < 0.0005), as was allele *11 (odds ratio 0.09, 95% CI 0.01-0.67; p < 0.007). Logistic regression gave similar results, showing that individuals with genotype 10/10 and 19-20 repeats (genotypes 9/10, 9/11, 10/10,) are protected from fracture or osteoporosis. Overall, there was a relationship between DBP Alu genotype and BMD, suggesting that DBP-Alu genotype may influence fracture risk. This effect may be mediated by changes in the circulating concentrations of DBP which influences free concentrations of vitamin D.

Objective evidence of cognitive complaints in Chronic Fatigue Syndrome: a BOLD fMRI study of verbal working memory.

Individuals with Chronic Fatigue Syndrome (CFS) often have difficulties with complex auditory information processing. In a series of two Blood Oxygen Level Dependent (BOLD) functional Magnetic Resonance Imaging (fMRI) studies, we compared BOLD signal changes between Controls and individuals with CFS who had documented difficulties in complex auditory information processing (Study 1) and those who did not (Study 2) in response to performance on a simple auditory monitoring and a complex auditory information processing task (mPASAT). We hypothesized that under conditions of cognitive challenge: (1) individuals with CFS who have auditory information processing difficulties will utilize frontal and parietal brain regions to a greater extent than Controls and (2) these differences will be maintained even when objective difficulties in this domain are controlled for. Using blocked design fMRI paradigms in both studies, we first presented the auditory monitoring task followed by the mPASAT. Within and between regions of interest (ROI), group analyses were performed for both studies with statistical parametric mapping (SPM99). Findings showed that individuals with CFS are able to process challenging auditory information as accurately as Controls but utilize more extensive regions of the network associated with the verbal WM system. Individuals with CFS appear to have to exert greater effort to process auditory information as effectively as demographically similar healthy adults. Our findings provide objective evidence for the subjective experience of cognitive difficulties in individuals with CFS.

Real-time assessment of postprandial fat storage in liver and skeletal muscle in health and type 2 diabetes.

Liver and skeletal muscle triglyceride stores are elevated in type 2 diabetes and correlate with insulin resistance. As postprandial handling of dietary fat may be a critical determinant of tissue triglyceride levels, we quantified postprandial fat storage in normal and type 2 diabetes subjects. Healthy volunteers (n = 8) and diet-controlled type 2 diabetes subjects (n = 12) were studied using a novel 13C magnetic resonance spectroscopy protocol to measure the postprandial increment in liver and skeletal muscle triglyceride following ingestion of 13C-labeled fatty acids given with a standard mixed meal. The postprandial increment in hepatic triglyceride was rapid in both groups (peak increment controls: +7.3 +/- 1.5 mmol/l at 6 h, P = 0.002; peak increment diabetics: +10.8 +/- 3.4 mmol/l at 4 h, P = 0.009). The mean postprandial incremental AUC of hepatic 13C enrichment between the first and second meals (0 and 4 h) was significantly higher in the diabetes group (6.1 +/- 1.4 vs. 1.7 +/- 0.6 mmol x l(-1) x h(-1), P = 0.019). Postprandial increment in skeletal muscle triglyceride in the control group was small compared with the diabetic group, the mean 24-h postprandial incremental AUC being 0.2 +/- 0.3 vs. 1.7 +/- 0.4 mmol x l(-1) x h(-1) (P = 0.009). We conclude that the postprandial uptake of fatty acids by liver and skeletal muscle is increased in type 2 diabetes and may underlie the elevated tissue triglyceride stores and consequent insulin resistance.

Stress effect of antilymphocyte serum.

Antilymphocyte serum prepared in the rabbit against guinea-pig lymph node cells produces a sustained weight loss when injected into normal guinea-pigs over 14 days, compared with a very transient loss produced by normal rabbit serum. Antilymphocyte serum also produces a much greater increase in plasma cortisol level than normal rabbit serum. Part of this effect may be due to adrenal stimulation, and further studies of its activity in adrenalectomized animals are suggested.