Expression of prostate specific antigen on the surface of a filamentous phage.

We have constructed two phagemid vectors containing the gene coding for human Prostate Specific Antigen (PSA) translationally fused to a pelB signal sequence and minor coat protein III of phage fd leading to phage particles that carry PSA on their tips. Phages were characterized by Western blotting and by panning, using biotinylated monoclonal anti-PSA antibodies immobilized onto streptavidin-coated microtitration wells. Binding of PSA-phages was 10(2)- to 5 x 10(3)-fold more effective than that of wild-type phages.

Lipoprotein fractionation in deuterium oxide gradients: a procedure for evaluation of antioxidant binding and susceptibility to oxidation.

Oxidative modifications of lipoproteins appear to contribute to their atherogenicity. Very low and low density lipoproteins (VLDL and LDL) are protected against these modifications by antioxidants that can be incorporated in vivo or in vitro into the particles. We describe here ultracentrifugal procedures for isolation of VLDL and LDL that do not require subsequent dialysis or buffer equilibration.

A new simple and rapid dual assay for AFP and free beta hCG in screening for Down syndrome.

We have evaluated a simple and rapid 2-step dual-label assay (DELFIA) for alphafetoprotein (AFP) and free beta subunit of human gonadotropin (hCG beta) in second-trimester screening for Down syndrome. Based on stored serum samples from 1059 normal control pregnancies and 72 cases of Down syndrome, we have found the mean Multiple of Median (MoM) for AFP and free hCG beta to be 0.70 and 2.

Surgery for chronic symptoms after whiplash injury. Follow-up of 20 cases.

20 patients operated on with discectomy and anterior cervical fusion because of chronic symptoms after whiplash injury were examined 4 years after surgery. The commonest indications for surgery were disabling headache and neck pain and radiographically verified disc protrusion. At follow-up, 11 patients had reduced headache and neck pain, while paresthesia and radicular pain were diminished in 9 patients.

Simultaneous assay of alpha-fetoprotein and free beta subunit of human chorionic gonadotropin by dual-label time-resolved immunofluorometric assay.

We developed a simple, rapid two-step dual-label assay for the noncompetitive determination of alpha-fetoprotein (AFP) and beta subunit of human chorionic gonadotropin (hCG beta) in serum. Monoclonal antibodies to detect AFP and hCG beta were labeled with europium (Eu) and samarium (Sm), respectively. Highly fluorescent chelates were developed by using the Delfia enhancement principle.

Concanavalin A affinity chromatography of human serum gonadotropins: evidence for changes of carbohydrate structure in different clinical conditions.

We have studied the carbohydrate of circulating human gonadotropins (FSH and LH) in different clinical conditions using Concanavalin A (Con A) affinity chromatography. This technique permits separation of molecules differing in the extent of carbohydrate branching. The proportion of molecules that does not bind to Con A was greater in circulating FSH than in LH, reflecting a higher content of multiantennary and/or bisected biantennary complex carbohydrate structures in serum FSH.

A supersensitive immunofluorometric assay for rat luteinizing hormone.

We have developed an immunofluorometric assay (IFMA) for rat (r) LH, which is based on two monoclonal antibodies, one to bovine and the other to human LH. Signal detection occurs by time-resolved fluorescence evoked by a europium label (Delfia, Wallac). The method is fast in comparison to the standard RIA with the NIDDK reagents (4 h vs.

Determination of dextropropoxyphene and norpropoxyphene in plasma by high-performance liquid chromatography.

A simple and sensitive procedure for the routine assay of the analgesic drug dextropropoxyphene and its main metabolite, norpropoxyphene, in plasma is described. After liquid-liquid extraction from alkalinized plasma and back-extraction into a small volume of an acidic aqueous phase, the aqueous phase was injected into a column packed with 3-microns octadecylsilica particles. Ultraviolet absorbance detection at 210 nm was used.

Inhibition of platelet accumulation by beta 1-adrenoceptor blockade in the thoracic aorta of rabbits subjected to experimental sympathetic activation.

Arterial platelet adhesion is an initiating event in the thrombo-embolic complications of atherosclerosis and may also accelerate the development rate of atherosclerotic lesions. Psychosocial stress has been shown to accelerate atherogenesis in animals, an effect probably mediated via beta-adrenoceptor activation. In view of the postulated roles of platelets and beta-adrenoceptor activation in atherosclerosis development, we decided to test whether beta blockade affects arterial platelet accumulation.

Simultaneous quadruple-label fluorometric immunoassay of thyroid-stimulating hormone, 17 alpha-hydroxyprogesterone, immunoreactive trypsin, and creatine kinase MM isoenzyme in dried blood spots.

We describe a quadruple-label fluorometric immunoassay for simultaneously measuring four analytes: thyroid-stimulating hormone (TSH), 17 alpha-hydroxyprogesterone (17 alpha-OHP), immunoreactive trypsin (IRT), and creatine kinase MM (CK-MM). The assay is based on immunoreagents labeled with four different lanthanide ions (Eu3+, Tb3+, Sm3+, and Dy3+), on dissociative fluorescence enhancement applying the principle of co-fluorescence, and on time-resolved fluorometry. The monoclonal anti-alpha-TSH and anti-IRT antibodies and the polyclonal anti-CK-MM antibody were labeled with Eu3+, Sm3+, and Dy3+, respectively; 17 alpha-OHP was labeled with Tb3+.

An immunologically anomalous luteinizing hormone variant in a healthy woman.

An investigation was undertaken to characterize an immunological LH variant in a 31-yr-old healthy woman whose serum LH was either poorly or not at all recognized by two monoclonal antibodies. The two antibodies recognize epitopes present on the intact LH dimer, but not on the free subunits. It was found that the immunologically aberrant LH of the subject was bioactive, as evidenced by an in vitro bioassay for LH.

Prostacyclin synthesis in relation to sympathoadrenal activation. Effects of beta-blockade.

Results from several studies suggest that beta-adrenoceptor blockade causes increased biosynthesis of the vasodilator prostanoid prostacyclin in the arterial wall. This effect may contribute to the clinical effects of beta-blockers in hypertension and coronary heart disease. Studies in hypertensive patients and in animals indicate that the arterial pressure reduction after beta-blockade is related to the associated increase of prostacyclin biosynthesis, regarding both magnitude and time of onset of effect.

Role of sympathoadrenal medullary activation in the initiation and progression of atherosclerosis.

Clinical and epidemiological investigations provide evidence that psychosocial factors influence the development of coronary heart disease and underlying atherosclerosis, an association that appears to be independent of the effects of other coronary disease risk factors. It has been hypothesized that sympathoadrenal medullary activation mediates behavioral influences on coronary disease, perhaps by potentiation of atherogenesis. This article summarizes four recent studies of the effects of psychosocial stress and sympathetic arousal on atherogenesis in cynomolgus monkeys (Macaca fascicularis) and rabbits.

Monoclonal antibody-based discrepancies between two-site immunometric tests for lutropin.

We compared five two-site immunometric assays (including three commercially available kits) for measuring lutropin (LH) in serum. Four of the assays involved monoclonal antibodies directed against the alpha-beta dimer, intact LH; these assays measured significantly lower concentrations of LH in 19 (out of 83) samples than did a commercial method not involving such antibodies. In five serum samples, two of the intact LH assays failed to detect any significant immunoreactivity above the detection limit.

Prostate-specific antigen in serum occurs predominantly in complex with alpha 1-antichymotrypsin.

Immunologic measurements of the serum concentration of prostate-specific antigen (PSA), an abundant prostatic-secreted serine proteinase, are frequently used to monitor patients with prostate cancer, though it has not been ascertained whether this immunoreactivity represents a PSA zymogen, the active proteinase, or PSA complexed to extracellular proteinase inhibitors. To characterize the PSA immunoreactivity in serum, we used monoclonal antibodies produced against PSA and a polyclonal rabbit IgG against alpha 1-antichymotrypsin in the design of three noncompetitive PSA assays: assay T, which detected PSA both when present as the active proteinase and when complexed to alpha 1-antichymotrypsin; assay F, which recognized the active proteinase but most poorly detected PSA complexed to alpha 1-antichymotrypsin; and assay C, which was specific for PSA complexed to alpha 1-antichymotrypsin. We used the three assays to measure PSA immunoreactivity in 64 patients' sera and in the effluent after gel chromatography of sera from four patients.

Effect of felodipine on left ventricular performance in conscious dogs: assessment by left ventricular pressure-volume analysis.

We assessed the effect of felodipine on left ventricular (LV) performance in 10 healthy, conscious, chronically instrumented dogs, using load-independent measures derived from variably loaded pressure-volume relations. With all reflexes intact, felodipine (25 nmol/kg, intravenously) caused significant decreases in LV end-systolic pressure (132 +/- 13 vs. 109 +/- 14 mm Hg, P less than .

Individual differences in lutropin immunoreactivity revealed by monoclonal antibodies.

We screened 11 monoclonal anti-lutropin (LH) antibodies for use in two-site immunometric combinations, with time-resolved fluorescence as the detection principle. Seven immunofluorometric assays (IFMA), six detecting only the intact hormone and one detecting both the intact hormone and free beta subunit, were compared with each other and with a conventional radioimmunoassay involving a polyclonal antiserum. Two monoclonal antibodies that exclusively reacted with intact LH showed restricted reactivity with LH in 25% of the samples tested.

Liquid chromatographic determination of the enantiomers of ibuprofen in plasma using a chiral AGP column.

A reversed-phase high-performance liquid chromatographic method has been developed for the determination of the R- and S-enantiomers of ibuprofen. The enantiomers and the internal standard 4-pentylphenylacetic acid are extracted from plasma, separated and quantified on a Chiral-AGP column using ultraviolet detection. The simplicity, sensitivity and precision of the method makes it convenient for use in pharmacokinetic studies.

The effects of age, gender, and parity on rectoanal functions in adults.

The effects of age, gender, and parity on rectoanal function were examined in a cross-sectional population study, including 68 normal subjects (32 men and 36 women) aged 23 to 91 years. Rectal volumetry was evaluated with graded isobaric rectal distension with 5-60 cm H2O in combination with anal manometry. Data were analysed by use of the multiple regression technique.

Ultrasensitive two-site immunometric assay of human lutropin by time-resolved fluorometry.

A rapid, sensitive two-site immunometric assay for human lutropin (LH), with two monoclonal antibodies directed against the beta subunit, has been developed with time-resolved fluorescence as the detection principle. The cross-reactivities with human thyrotropin and human follitropin are negligible and that with human choriogonadotropin is 1% in the standard two-step protocol. Free beta subunits of LH are also detected.

Experimental sympathetic activation causes endothelial injury in the rabbit thoracic aorta via beta 1-adrenoceptor activation.

Sympathetic activation appears to accelerate the development of atherosclerosis, an effect that may be inhibited by beta-receptor blockade. It is unclear, however, which mechanisms mediate this effect. In view of the significance attached to endothelial injury in the initial phases of atherogenesis, we decided to test whether sympathetic activation might lead to an increase in endothelial injury.