Serum concentrations of retinol, d-alpha-tocopherol and beta-carotene: effects of storage at -70 degrees C for five years

To investigate the effects of prolonged storage of serum samples at -70 degrees C on concentrations of micronutrients, we measured concentrations of retinol, d-alpha-tocopherol, and beta-carotene in serum samples drawn in 1986. We compared values we measured in 1991 to values we obtained in 1986, using the same analytical methods. The relative concentrations obtained in 1991 (mean +/- S.D.) were: retinol 99.7 +/- 12.6% (n = 23), d-alpha-tocopherol 100.7 +/- 6.4% (n = 19), and beta-carotene 103.4 +/- 13.7% (n = 28). Using these techniques of sample preparation and high-performance liquid chromatographic analysis, we found that the effects of storage of serum at -70 degrees C for five years appear insignificant in a small population of patients. However, we did identify clinically important changes in concentration (> 20% difference) in several individual subjects.     

Effects of fasting and 3,3',4,4',5,5'-hexabromobiphenyl on plasma transport of thyroxine and retinol: fasting reverses elevation of retinol

Male Wistar rats were injected ip with 0 or 20 mg/kg 3,3',4,4',5,5'-hexabromobiphenyl and blood samples were collected 1, 3, 6, 7, and 8 d later. At 8 d after the injection, serum retinol was increased 30% and serum thyroxine was decreased 26% relative to control values. These effects were apparently unrelated to transthyretin in that the biphenyl did not alter the proportion of thyroxine binding in vitro to the prealbumin fraction of serum proteins. Separate groups of control and HBBP-injected rats did not receive food on d 7 (i.e., 24-h fast) and d 8 after injection (i.e., 48-h fast). Fasting decreased the serum retinol and thyroxine concentrations as well as the proportion of thyroxine binding in vitro to the prealbumin fraction of serum. The decreases in retinol and thyroxine concentrations associated with fasting are therefore ascribed to a decrease in the concentration of transthyretin in circulation.     

A critical role for transforming growth factor-beta in donor transfusion-induced allograft tolerance

To evaluate the relationship between carotenoid concentrations in serum and breast tissue, we measured serum carotenoid concentrations and endogenous carotenoid levels in breast adipose tissue of women with benign breast tumor (n = 46) or breast cancer (n = 44). Before extraction, serum was digested with lipase and cholesterol esterase, and breast adipose tissue was saponified. Serum and tissue carotenoids were extracted with ether/hexane and measured by using HPLC with a C30 column. Serum retinoic acid was extracted with chloroform/methanol and measured using HPLC with a C18 column. There were no significant differences in serum carotenoids [lutein, zeaxanthin, cryptoxanthin (both alpha- and beta-), alpha-carotene, all-trans beta-carotene, 13-cis beta-carotene and lycopene], retinoids (retinol, all-trans and 13-cis retinoic acids), and alpha- and gamma- tocopherol concentrations between benign breast tumor patients and breast cancer patients. A substantial amount of 9-cis beta-carotene was present in adipose tissue and was the only carotenoid that had a significantly lower level in benign breast tumor patients than in breast cancer patients. Correlations between carotenoid concentrations in serum and in breast adipose tissue were determined by combining the data of the two groups. Concentrations of the major serum carotenoids except cryptoxanthin showed significant correlations with breast adipose tissue carotenoid levels. When the concentrations of serum carotenoids were adjusted for serum triglycerides or LDL, correlations between serum carotenoid concentrations and breast adipose tissue carotenoid levels markedly increased, including that of cryptoxanthin (P <0. 001). The strong correlation between serum carotenoid concentrations and endogenous breast adipose tissue carotenoid levels indicate that dietary intake influences adipose tissue carotenoid levels as well as serum concentrations, and that adipose tissue is a dynamic reservoir of fat-soluble nutrients.     

Single dose pharmacokinetics of manidipine in hepatic impaired patients and healthy controls

The pharmacokinetics and safety of a single oral dose of 20 mg manidipine dihydrochloride have been studied in 8 patients with mild to moderate hepatic impairment (grade A or B in Child's classification, or score < or = 7 in Pugh's modification of Child's classification), and in 12 healthy subjects. They received one 20 mg manidipine dihydrochloride tablet with 100 ml of tap water after a standard breakfast. Manidipine was determined using HPLC with electrochemical detection from plasma samples taken up to 24 or 36 h after dosing. The medication was well tolerated. A trend toward higher Cmax, AUC, and MRT was observed in patients with a more severe hepatic impairment, as a consequence of reduction in the liver metabolic function. Patients with grade A hepatic impairment did not exhibit significantly altered pharmacokinetics with respect to healthy subjects, while grade B impairment patients had significantly higher AUC and MRT. Tmax values pointed to reduced absorption rate in patients compared to healthy subjects; the changes were more evident in grade B than grade A patients, although statistical significance was not reached. The reduction in absorption rate in grade B patients is probably related to their higher mean age, since this effect has been reported for manidipine. The pharmacokinetics of manidipine seem only modified in patients with a certain degree of hepatic impairment (at least Pugh grade 6 and Child grade B); therefore, adaptation of the dosing regimen does not seem to be generally recommendable, but should be modulated according to the liver status of the patient.     

Predictors of serum retinol in children with shigellosis

BACKGROUND: Low serum retinol can be useful as an indicator of depleted liver vitamin A stores, particularly in population-based studies. However, serum retinol concentrations decrease transiently during infection, independent of any changes in liver stores. The magnitude of the decrease in serum retinol is often proportional to indicators of disease severity. OBJECTIVE: We examined the relation of serum retinol in children with culture-positive shigellosis with severity of illness, anthropometric indicators of nutritional status, urinary retinol excretion, and serum concentrations of C-reactive protein, alpha1-acid glycoprotein, retinol binding protein, and transthyretin. DESIGN: This was a prospective study assessing the clinical and laboratory measurements at admission and recovery of 90 children with dysentery (66 with shigellosis) hospitalized in Bangladesh. RESULTS: Serum retinol concentrations were low at admission but were significantly greater at discharge even though no vitamin A supplements were given during the illness (0.36 +/- 0.22 compared with 1.15 +/- 0.50 micromol/L, P < 0.001). Serum retinol concentrations were lower in children with Shigella dysenteriae type 1 infection than in children with shigellosis due to less virulent strains of Shigella. Low serum retinol was independently associated with S. dysenteriae type 1, high serum C-reactive protein concentrations, and low weight-forage in multiple regression analysis. CONCLUSIONS: This study showed that shigellosis was associated with a significant, transient decrease in serum retinol concentrations of approximately 0.8 micromol/L, and that this change was significantly associated with severity of disease and poor underlying nutritional status, particularly low weight-for-age.     

Severity of respiratory syncytial virus disease related to type and genotype of virus and to cytokine values in nasopharyngeal secretions

BACKGROUND: Investigations concerning the severity of respiratory syncytial virus (RSV) disease as related to (1) RSV type and genotype determined respectively by PCR and restriction enzyme analysis and (2) interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) values in samples of nasopharyngeal secretion (NPS) have not been previously reported. METHODS: We prospectively studied 105 RSV infections in the lower respiratory tract of infants and young children admitted to a pediatric department in Copenhagen during three winter seasons, 1993, 1994 and 1995. RSV strains were typed and genotyped, respectively, by PCR and nucleic acid restriction analysis and correlated to the severity of the disease. The ratio IL-6:TNF-alpha, determined from IL-6- and TNF-alpha values in samples of NPS, was related to the severity of the disease. Concentrations of IL-6 and of TNF-alpha were determined in serum samples taken during 5 weeks after the onset of illness. RESULTS: Type B infections produced more severe disease than did type A infections, as assessed on the length of the hospital stay, use of respiratory support and the presence of an infiltrate on a chest radiograph. This difference was age-related. It was observed in infants 0 to 5 months old, but not in older age groups. Type B genotype B1122 produced more severe disease than type A genotype A2311 in infants 0 to 11 months old. Increased serum concentrations of IL-6 and TNF-alpha were detected in samples taken 1 to 2 days after the onset of illness. Whereas TNF-alpha serum concentrations remained high, IL-6 serum concentrations decreased during the following 3 to 4 weeks. The IL-6:TNF-alpha ratio in samples of NPS was related to the severity of the disease. A high ratio was related to a low severity. CONCLUSIONS: The severity of disease in patients admitted with acute RSV infections can be correlated to the RSV type as determined by PCR, to the RSV genotype as determined by nucleic acid restriction analysis and to the ratio IL-6:TNF-alpha in NPS.     

Cerebrospinal fluid levels of alpha-tocopherol in amyotrophic lateral sclerosis

We compared CSF and serum levels, and the CSF/serum ratio of alpha-tocopherol (vitamin E), measured by HPLC, in 30 patients with sporadic amyotrophic lateral sclerosis (SALS) and 78 matched controls. The mean CSF and serum vitamin E levels did not differ significantly between the 2 study groups. These values were not influenced by the clinical form (spinal versus bulbar) of SALS. CSF alpha-tocopherol levels did not correlate with age, age at onset, and duration of the disease. These results suggest that CSF and serum alpha-tocopherol concentrations are unrelated with the risk for ALS.     

Sarcoidosis

Total serum immunoglobulin E (IgE) concentrations were determined by a competitive solid phase radioimmunoassay technique in serum samples from patients with a variety of venereal diseases. The mean IgE concentrations for groups of normal persons without venereal diseases was significantly lower then the means for groups of appropriately matched patients with primary syphilis and gonorrhoea. There were also relatively higher IgE values in patients with trichomoniasis. Our data indicate that patients with urogenital infections have higher concentrations of IgE in the serum than matched control patients without such infections.     

Development of an automated turbidimetric immunoassay for quantification of bovine serum immunoglobulin G

OBJECTIVE: To develop an automated turbidimetric immunoassay (TIA) for measurement of bovine IgG. SAMPLE POPULATION: 24 bovine serum samples. PROCEDURE: IgG concentration was determined by use of the TIA, and results were compared with those of the radial immunodiffusion (RID) method. Variables were determined, using commercially available reagents and a clinical biochemical analyzer. For the TIA, polyclonal goat anti-bovine IgG (Fc specific) serum, bovine IgG calibrator serum, and polyethylene glycol reaction buffer were used. Sample concentrations were determined by the instrument, using the linear regression method of least squares. The accuracy of this assay was validated by referencing to a purified bovine IgG standard and by recovery of control standards. Parallelism was documented by assay linearity and serial sample dilution linearity. Interference resulting from hemolyzed samples was examined. RESULTS: The TIA method correlated positively (r = 0.9957) and significantly (P < 0.05) with the RID method, yielding a regression equation with slope of 0.78708 and y-intercept of 1.02102. Bias attributable to hemolysis was not observed. CONCLUSIONS: The TIA method is automated, accurate, and precise for bovine serum IgG quantification. CLINICAL RELEVANCE: This assay provides sample results in approximately 10 minutes and may be used as an alternative to the manual RID method.     

Surface charge control of electropermeabilization and glycophorin electroinsertion with 1,2-diacyl-sn-glycero-3-phosphocholine (lecithin) liposomes

beta-Carotene has been studied widely as a potential cancer-preventing agent. Recent studies found that subjects who took beta-carotene supplements orally had increases in their serum concentrations of alpha-carotene and lycopene that were large (> 150% increase) and significantly greater than such increases in subjects who received placebo and that similar supplementation was associated with a decrease of approximately 37% in plasma lutein concentrations. A biologic interaction between beta-carotene and other carotenoids was suggested. We measured concentrations of retinol, alpha-tocopherol, and five carotenoids in serum specimens from a random sample of subjects enrolled in a clinical trial of the use of antioxidant vitamins in preventing colonic adenomas. We used serum specimens obtained at enrollment and after the subjects took placebo (n = 54) or 25 mg beta-carotene/d (n = 54) orally for 4 y. In a multivariate analysis, baseline serum concentrations of the analytes, sex, body mass index, diet, smoking status, and age were associated with variable changes in some analytes over the 4-y period but supplementation with beta-carotene was related only to a mean increase in serum beta-carotene itself of 151%. We excluded with 95% confidence an increase in lycopene > 4.9%, an increase in alpha-carotene > 17.6%, and a decrease in lutein > 14.7% in subjects given beta-carotene. These results confirm previous findings that supplementation with beta-carotene given orally does not alter serum concentrations of retinol or alpha-tocopherol. The findings also indicate that beta-carotene supplementation, which results in a moderate increase in serum beta-carotene concentration, does not significantly change serum concentrations of other carotenoids.     

Validation of procedures for quantitative whole-body autoradiography using digital imaging

The objective of this study was to demonstrate that tissue concentrations of radioactivity derived by digital analysis of autoradiograms were comparable to values derived from direct sampling and analysis of tissues. In addition, we describe the preparation and calibration of standards for use in quantitative whole-body autoradiography. For this study, three male Long-Evans hooded rats were administered 14C radioactivity intravenously. The animals were sectioned for whole-body autoradiography, with concomitant sampling of blood and 16 selected tissues. After 3 weeks of film exposure, the optical densities of the resulting autoradiograms were analyzed with a RAS3000 digital imaging system to estimate tissue concentrations of radioactivity. These concentrations were then compared with those obtained by direct analysis of the tissue samples. The concentrations derived from digital analysis of the autoradiograms were very highly correlated with those determined from direct tissue analysis (r = 0.956). Linear regression analysis yielded a straight line with a slope of 0.97 and a goodness of fit (r2) of 0.913. This analysis suggested that there is an approximate 1:1 correlation between concentration values determined by the two methods. Marked differences between the values derived via the two techniques were observed for only three tissues. However, this subset of the data accounted for only 6% of the total data, and the differences were probably due to contamination from adjacent tissues during excision. Overall, the concentrations of radioactivity derived from digital analysis of the autoradiograms were comparable to those derived from direct analysis of tissue samples. The results indicated that the digital analysis procedure for film can serve as a valuable adjunct to conventional tissue analysis for radioactivity.     

Reappraisal of the role of vegetables in the vitamin A status of mothers in Central Java, Indonesia

Food-based approaches for controlling vitamin A deficiency and its consequences, such as increased mortality, more severe morbidity, and anemia, have become increasingly important, thus prompting a reassessment of the relation between vitamin A intake and status. A nutrition surveillance system in Central Java, Indonesia, assessed the vitamin A intake and serum retinol concentration of women with a child < or =24 mo old with a semiquantitative 24-h recall method that categorized vitamin A-containing foods into 3 categories of plant foods and into 2 categories of animal foods and identified portions as small, medium, or large. Median vitamin A intake was 335 retinol equivalents (RE)/d (n = 600) and vitamin A intake from plant foods was 8 times higher than from animal foods. Serum retinol concentration was related to vitamin A intake in a dose-response manner. The multiple logistic regression model for predicting the chance for a serum retinol concentration greater than the observed median (> or = 1.37 micromol/L) included physiologic factors, vitamin A intake from plant [odds ratio (95% CI) per quartile: 1st, 1.00: 2nd, 1.23 (0.75, 2.02); 3rd, 1.60 (0.97, 2.63); and 4th, 2.06 (1.25, 3.40)] and animal [1st and 2nd, 1.00; 3rd, 1.31 (0.86, 2.02); and 4th, 2.18 (1.40. 3.42)] foods, home gardening [(no, 1.00; yes, 1.71 (1.12, 2.60)], and woman's education level [< or =primary school, 1.00; > or =secondary school, 1.51 (1.02, 2.22)]. Despite the fact that plant foods contributed 8 times as much vitamin A as did animal foods, serum retinol concentrations did not reflect this large difference. Home gardening and woman's education level seemed to reflect longer-term consumption of vitamin A-rich plant and animal foods, respectively.     

A routine method for the simultaneous measurement of retinol, alpha-tocopherol and five carotenoids in human plasma by reverse phase HPLC

We describe a simple isocratic HPLC method for the accurate and precise measurement of retinol, alpha-tocopherol and the major carotenoids in plasma using UV detection. Reference ranges for retinol, alpha-tocopherol and five carotenoids are determined in a healthy population group. The most abundant carotenoids found in plasma were beta-carotene, lycopene, lutein and cryptoxanthin. Retinol, alpha-tocopherol and carotenoids were determined simultaneously using two internal standards, retinol acetate for retinol and tocopherol acetate for alpha-tocopherol and carotenoids. The use of echinenone as an internal standard for carotenoids was investigated. The protective effect of an antioxidant (ascorbic acid) on the stability of samples and extracted material is documented. The method is useful for the routine measurement of plasma retinol, alpha-tocopherol and carotenoids and could also be used in large scale epidemiological studies.     

Developmental pattern of fetal growth hormone, insulin-like growth factor I, growth hormone binding protein and insulin-like growth factor binding protein-3

AIMS: To evaluate the developmental pattern of fetal growth hormone (GH), insulin-like growth factor I (IGF-I), GH binding protein (GHBP) and IGF binding protein-3 (IGF-3); to determine the implications for fetal growth. METHODS: Serum GH, IGF-I, GHBP and IGFBP-3 were measured in 53 fetuses, 41 aged 20-26 weeks (group A) and 12 aged 31-38 weeks (group B). Fetal blood samples were obtained by direct puncture of the umbilical vein in utero. Fetal blood samples were taken to rule out beta thalassaemia, chromosome alterations, mother to fetus transmissible infections, and for maternal rhesus factor. GHBP was determined by gel filtration chromatography of serum incubated overnight with 125I-GH. GH, IGF-I and IGFBP-3 were determined by radioimmunoassay. RESULTS: Fetal serum GH concentrations in group A (median 29 micrograms/l, range 11-92) were significantly higher (P < 0.01) than those of group B (median 16.7 micrograms/l, range 4.5-29). IGF-I in group A (median 20 micrograms/l, range 4.1-53.3) was significantly lower (P < 0.01) than in group B (median 75.2 micrograms/l, range 27.8-122.3). Similarly, IGFBP-3 concentrations in group A (median 950 micrograms/l, range 580-1260) were significantly lower than those of group B (median 1920 micrograms/l, range 1070-1770). There was no significant difference between GHBP values in group A (median 8.6%, range 6.6-12.6) and group B (median 8.3%, range 6-14.3). Gestational age correlated positively with IGF-I concentrations (P < 0.0001) and IGFBP-3 (P < 0.0001) and negatively with GH (P < 0.0001). GHBP values did not correlate with gestational age. Multiple regression analysis showed a negative correlation between GH:IGF-I ratio and fetal growth indices CONCLUSIONS: The simultaneous evaluation of fetal GH, IGF-I, IGFBP-3 and GHBP suggests that the GH-IGF-I axis might already be functional in utero. The progressive improvement in the efficiency of this axis in the last part of gestation does not seem to be due to an increase in GH receptors.     

Occurrence of fumonisins in corn-based food products

Corn-based food products obtained from commercial outlets in three different parts of the U.S., Maryland, Nebraska, and Arizona were analyzed for total fumonisins by a commercial competitive direct enzyme-linked immunosorbent assay (CD-ELISA) and for fumonisin B1 (FB1) by high-performance liquid chromatography (HPLC). The highest fumonisin concentrations were found in samples collected in Maryland, where all 18 samples were found positive for fumonisins (200 to 7,450 ng/g of food) by CD-ELISA and 15 of the 18 samples (83%) were found positive for FB1 (< 75 to 5,916 ng/g) by HPLC. Fumonisins were also detected by CD-ELISA in 14 of 15 samples collected in Arizona with concentrations ranging from 200 to 1,450 ng/g, but analyses by HPLC showed that only 8 of 15 samples (53%) were positive for FB1 (< 75 to 1,565 ng/g of food). Of the 23 samples collected in Nebraska, 20 (87%) were positive for fumonisins (200 to 2,500 ng/g) by CD-ELISA, but only 10 (44%) were positive for FB1 (< 75 to 927 ng/g) by HPLC. The highest fumonisin and FB1 concentrations were found in cornmeal samples, ranging up to 7,450 ng/g of cornmeal by CD-ELISA and 5,916 ng/g by HPLC. These findings indicate that there may be a risk of human exposure to fumonisins through the consumption of some corn-based foods.     

Synthesis and QSAR of some 3-amino-2-(substituted)aminomethyl-5,6-disubstituted thieno[2,3-d]pyrimidin-4(3H)-ones as novel H1-receptor antagonists

The plasma concentrations of two bone matrix proteins (osteocalcin, osteonectin) were monitored in 56 samples from 14 patients receiving renal transplants and the values compared with serum bone alkaline phosphatase mass concentrations and osteotropic hormone levels (parathyroid hormone, calcitriol). There were no significant changes in the concentrations of plasma osteonectin at any time after transplantation, as compared with the values before transplantation (P > 0.1). None of the plasma samples showed osteonectin levels above the reference interval. There was a weak but significant relationship between platelet counts and plasma osteonectin levels (r = +0.322; P < 0.05). Osteocalcin showed a marked decrease of the values 1 week following transplantation as compared with the values before transplantation without further change of the values 1 and 3 months after transplantation (P > 0.5) whereas 3 months after transplantation bone alkaline phosphatase levels were higher than before transplantation (P < 0.05). Multiple regression analysis (performed with data from 42 samples obtained after transplantation) revealed serum creatinine as an independent predictor of plasma osteocalcin whereas serum calcitriol was an independent predictor of serum bone alkaline phosphatase (P < 0.05). No correlation was observed between serum calcitriol/plasma parathyroid hormone on the one hand and plasma osteocalcin on the other (P > 0.05). After transplantation there was a lack of correlation between serum bone alkaline phosphatase mass concentrations and plasma osteocalcin values (P > 0.05). In conclusion, serum bone alkaline phosphatase should be preferred to bone matrix proteins for the assessment of bone metabolism in patients receiving renal transplants: (a) bone alkaline phosphatase-but not osteocalcin-is significantly correlated with calcitriol and adequately reflects increased bone formation after renal transplantation; (b) interpretation of osteocalcin values is severely hampered by their strong correlation with serum creatinine concentrations; (c) plasma osteonectin determinations are not useful for monitoring bone formation.     

Serum beta-carotene and antioxidant micronutrients in children with cancer. The 'Cancer in Children and Antioxidant Micronutrients' French Study Group

Serum antioxidant vitamins A (retinol) and E (alpha-tocopherol), beta-carotene, zinc and selenium for 418 children with newly diagnosed malignancy were compared with those of 632 cancer-free controls. Incident cancer cases and controls were 1-16 years old and recruited in 1986-1989. Age- and sex-adjusted serum concentrations of retinol, beta-carotene and alpha-tocopherol were significantly inversely associated with cancer. In similar models, the odds ratio (OR) comparing the highest with the lowest quintile was 2.06 (95% confidence interval [CI] 1.40-3.02) for retinol, 3.87 (95% CI: 2.54-5.90) for beta-carotene, 2.15 (95% CI: 1.48-3.10) for alpha-tocopherol, 1.29 (95% CI: 0.75-2.23) for selenium, and 1.94 (95% CI: 1.17-2.23) for zinc. The cancer sites that were associated with serum beta-carotene were, in general, leukaemia, lymphoma, central nervous system, bone and renal tumours. Moreover, leukaemia was associated with low mean serum levels of retinol, selenium and zinc. Subjects with lymphoma, bone and renal tumours also had lower mean retinol and alpha-tocopherol levels than controls. Brain tumour patients had low vitamin E levels. Low serum values of antioxidant vitamins were associated with childhood neoplasm occurrence. Some site-specific effect was reported. Low peripheral nutrient levels are not considered as cancer promoters but rather as an impairment of the body's defence mechanism occurring during the cancer-related metabolic and nutritional disturbances and inflammation processes.     

Alterations in acid-base balance and serum electrolyte concentrations in cattle: 632 cases (1984-1994)

OBJECTIVE: To determine typical alterations in acid-base balance and serum electrolyte concentrations in cattle grouped on the basis of age, breed type, hydration status, clinical signs, and underlying disease. DESIGN: Retrospective study. ANIMALS: 632 cattle. PROCEDURE: Information on blood pH, PVO2, PVCO2, HCO3-concentration, and base excess (BE) as well as serum or plasma sodium, potassium, chloride, and total carbon dioxide concentrations was obtained. Values for calves (ie, cattle < 1 month old) were compared with values for cattle > or = 1 month old. Within each age-group, values were compared for cattle grouped on the basis of breed type, previous treatment, and hydration status. Proportions of cattle with various disorders for which values were within, greater than, or less than reference ranges were determined. RESULTS: BE, pH, and HCO3- concentration were significantly higher and PVCO2 and sodium, potassium, and chloride concentrations were significantly lower among cattle > or = 1 month old than calves. Base excess and HCO3- concentration were significantly lower among dairy than beef calves. Sodium and potassium concentrations were significantly lower among dairy than beef cattle > or = 1 month old. A higher proportion of cattle that did not survive had a high anion gap than cattle that did survive. Sodium, potassium, and chloride concentrations were significantly lower among dehydrated cattle than cattle that were not dehydrated. CLINICAL IMPLICATIONS: Because certain alterations were consistently found in some groups of cattle, it may be possible to make reasonable predictions of alterations in acid-base balance and serum electrolyte concentrations when laboratory evaluations are not available.     

Health sector reform: priorities and packages

Using time-resolved fluorometry, a simple one-step dual-label immunometric assay has been developed, which allows simultaneous determination of pregnancy-associated plasma protein A (PAPP-A) and free beta-subunit of human chorionic gonadotrophin (beta-hCG) in first-trimester maternal serum samples. Two monoclonal antibodies were biotinylated and immobilized onto the surface of streptavidin-coated microtitration plates, and used to capture PAPP-A and beta-hCG. respectively. Europium (Eu) and Samarium (Sm) chelates were conjugated to two additional monoclonal antibodies acting as detection antibodies for PAPP-A and beta-hCG. The assay was performed using a 4-h one-step format. The within-run precision with buffer-based calibrators was below 8% over the working range of PAPP-A (40-10000 mIU/l) and beta-hCG (7.3-525 micrograms/l) and no hook effect was observed. The intra- and inter-assay coefficients of variation were below 7.1% for serum samples. PAPP-A and beta-hCG concentrations measured by the dual assay in 39 first-trimester serum samples correlated excellently with those obtained by DELFIA single-label PAPP-A (r = 0.997) and the beta-hCG part (r = 0.993) of the DELFIA AFP/beta hCG dual-label assay.     

Monitoring serum concentrations of clomipramine and metabolites: fluorescence polarization immunoassay versus high performance liquid chromatography

This study compared fluorescence polarization immunoassay (FPIA) with a high performance liquid chromatographic (HPLC) method from the point of view of their applicability to therapeutic drug monitoring of patients treated with clomipramine alone. Blood was withdrawn from 20 depressed inpatients (54 +/- 14 years) under steady state conditions. The FPIA determined total tricyclic antidepressant (TCA) concentrations with day to day variability below 11%. The automated HPLC method separated clomipramine, N-desmethylclomipramine, 8-hydroxyclomipramine and 8-hydroxydesmethylclomipramine with interassay coefficients of variance below 12%. The concentrations measured by FPIA were similar to HPLC results. Total TCA concentrations measured by FPIA and the sum of clomipramine and desmethylclomipramine measured by HPLC correlated significantly (r = 0.780 and p < 0.01). However, 40% of individual FPIA determinations yielded results that differed by more than 50% from the HPLC concentrations. Changes in clinical rates were related only to TCA serum concentrations that had been analyzed by HPLC. It is concluded that the semiquantitative FPIA is unsuitable for therapeutic drug monitoring in patients under clomipramine treatment, whereas the differential analysis of clomipramine and metabolites by HPLC is informative and can be used to improve the antidepressant drug treatment.