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PubMed1999

Macroprolactinaemia: validation and application of the polyethylene glycol precipitation test and clinical characterization of the condition.

OBJECTIVE: The clinical characteristics and natural history of macroprolactinaemia are yet to be fully established and there is currently no widely accepted simple in vitro test for routine identification of the condition. This study was designed to further validate the polyethylene glycol (PEG) precipitation test with assessment of percentage prolactin recovery (%R), for the detection of macroprolactinaemia and clinically characterize cases identified using the test. DESIGN: Analytical validation and application of the PEG precipitation test for diagnosis of macroprolactinaemia and a retrospective case-notes review of some of the patients identified with the condition. PATIENTS: Sera from 188 patients with elevated serum prolactin (PRL) concentrations were screened for macroprolactinaemia. Seventeen of the patients who were under follow-up at the local teaching hospital and were identified to have macroprolactinaemia were systematically characterized clinically. MEASUREMENTS: Prolactin was measured in sera, fractions from gel filtration chromatography and supernatant obtained after PEG precipitation, by the DELFIA fluoroimmunoassay. CLINICAL ENDPOINTS: Initial serum PRL and symptoms and their response to dopamine agonist therapy; imaging of the pituitary gland; serum PRL and symptoms after cessation of dopamine agonist therapy; course of serum PRL and symptoms without dopamine agonist therapy. RESULTS: The within-batch coefficient of variation (CV) of the PEG precipitation test ranged between 2.8 and 7.3% while the between-batch CV ranged between 3.4 and 9.7%. Intra-individual variability up to five months was 9.9%. A prolactin recovery </= 40% was indicative of macroprolactinaemia, while the condition was unlikely to be present at values > 50%. The prevalence of macroprolactinaemia was 15% without sex difference. Clinical characterization of 17 patients showed modest initial serum PRL concentrations and there was imaging evidence of a coexisting pituitary adenoma in three of them. Dopamine agonist therapy resulted in substantial falls in serum PRL and this was associated with improvement or resolution of symptoms in some patients. Stoppage of dopamine agonist resulted in rebound hyperprolactinaemia in all cases. Spontaneous improvement in symptoms occurred in three patients but hyperprolactinaemia persisted in all those who were not on treatment. CONCLUSIONS: The PEG precipitation test with assessment of 'free' prolactin has been shown to be reproducible and sensitive for the detection of macroprolactinaemia. Many of the patients who had macroprolactinaemia presented with at least one symptom of the hyperprolactinaemic syndrome. Dopaminergic therapy may influence the course of both serum PRL and symptoms in some of these patients.

Adolescent↗

PubMed2003

Galectin-3 precipitates as a pentamer with synthetic multivalent carbohydrates and forms heterogeneous cross-linked complexes.

Galectin-3 is unique among the galectin family of animal lectins in its biological activities and structure. Most members of the galectin family including galectin-1 possess apoptotic activities, whereas galectin-3 possesses anti-apoptotic activity. Galectin-3 is also the only chimera type galectin and consists of a nonlectin N-terminal domain and a C-terminal carbohydrate-binding domain. Recent sedimentation equilibrium and velocity studies show that murine galectin-3 is a monomer in the absence and presence of LacNAc, a monovalent sugar. However, quantitative precipitation studies in the present report indicate that galectin-3 precipitates as a pentamer with a series of divalent pentasaccharides with terminal LacNAc residues. Furthermore, the kinetics of precipitation are fast, on the order of seconds. This indicates that although the majority of galectin-3 in solution is a monomer, a rapid equilibrium exists between the monomer and a small percentage of pentamer. The latter, in turn, precipitates with the divalent oligosaccharides, resulting in rapid conversion of monomer to pentamer by mass action equilibria. Mixed quantitative precipitation experiments and electron microscopy suggest that galectin-3 forms heterogenous, disorganized cross-linking complexes with the multivalent carbohydrates. This contrasts with galectin-1 and many plant lectins that form homogeneous, organized cross-linked complexes. The results are discussed in terms of the biological properties of galectin-3.

Carbohydrate Metabolism↗

PubMed1998

Delmopinol hydrochloride- and chlorhexidine digluconate-induced precipitation of salivary proteins of different molecular weights.

Gel electrophoresis was used to analyze precipitates formed of delmopinol hydrochloride or chlorhexidine digluconate mixed with unstimulated whole saliva samples from five test subjects. Final concentrations of delmopinol (6.4 mM) or chlorhexidine (6.4 mM, 2.2 mM) mixed with whole saliva were incubated for 10 min at 37 degrees C. The precipitates were pelleted by centrifugation and resuspended to a similar protein density. The protein patterns in the pellets were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, using 12.3% gels. The amount of pellet protein was determined by densitometry in four molecular weight ranges (10-21.5, 21.5-26, 26-45, and 45-300). The results indicated that high molecular weight (45-300) proteins dominated in the precipitate and that 2.2 mM chlorlhexidine precipitated more salivary protein than 6.4 mM. At equimolar concentration (6.4 mM) delmopinol precipitated more high molecular weight salivary proteins than chlorhexidine.

Adult↗

PubMed1976

Precipitation of plasma lipoproteins by PEG-6000 and its evaluation with electrophoresis and ultracentrifugation.

A neutral polymer precipitation procedure for fractionation of human plasma lipoproteins is described. The method was also tested preliminarily for fractionation of rat plasma lipoproteins. Polyethylene glycol-6000 (PEG-6000) at a concentration of about 12% and at pH 7.9 precipitated human plasma very low density (VLDL) and low-density (LDL) lipoproteins completely. Precipitation was independent of time and temperature. VLDL and LDL could not be precipitated separately. Polyethylene glycol-6000 precipitation can be used for rapid determination of high-density (HDL) lipids and of the ratio of HDL to LDL lipids.

Adolescent↗

PubMed2002

MAP precipitation from landfill leachate and seawater bittern waste.

The leachates generated at Hong Kong landfill sites contain high strength of ammonium-nitrogen (NH4+-N) over 3,000 mg l(-1) and are generally inhibitive to most biological treatment processes. To remove the NH4+-N from the landfill leachates and also recover the nitrogen as a struvite fertilizer, a lab-scale study was performed to investigate the feasibility of NH4+-N removal and struvite crystallization using different magnesium sources. Three combinations of chemicals, MgCl2 x 6H2O+Na2HPO4 x 12H2O, MgO+H3PO4 and MgSO4 x 7H2O+Ca(H2PO4) x H2O, were first used at different molar ratios to precipitate NH4+-N from the leachate. The experimental results indicated that NH4+-N was removed by 92%, 36% and 70% respectively at pH 9.0 and a molar ratio of Mg:N:P=1:1:1. Two synthetic seawater bitten wastes containing Mg2+ at 9,220 mg l(-1) and 24,900 mg l(-1) respectively were then used as a magnesium source, while 85% H3PO4 chemical was used as a phosphorus source. The results revealed that NH4+-N was removed by 80% and 72% respectively, while a molar ratio of Mg:N:P=1:1:1 was applied. In the final stage of experiments, the magnesium-ammonium-phosphate (MAP) precipitates were examined by SEM, EDS and XRD. The SEM micrographs of the MAP precipitates showed a typical morphology of elongated tubular and short prismatic crystals. The EDS and XRD results indicated that the chemical composition of the MAP precipitates depended on the chemicals used and experimental conditions. The study confirmed that the recovery of NH4+-N from landfill leachate and seawater bitten wastes could be effectively achieved by MAP precipitation to obtain struvite crystals with a composition of 5.1%N, 10.3%Mg and 16.5%P.

Chemical Precipitation↗

PubMed2005

[Study of the metal precipitation from decontamination leachates of municipal wastes fly ash incinerators].

This research work focuses on the development of a new process for the decontamination of municipal wastes incinerators fly ashes. The objective of this study was to evaluate different total and selective precipitation methods for metals removal from ash decontamination leachates. The tested options include 1) use of hydrated lime and caustic soda for selective (pH 5.0) and total (pH 8.5) metal precipitation; 2) addition of different chemicals (H3PO4, Na2S and FeCl3) in a pH range from 6.0 to 9.0. Fly ash decontamination assays using alkaline and acid washing steps were initially performed using optimal conditions previously established. Treated fly ashes respected the standards based on the TCLP leaching test for all studied metals and SPLP. Total metal precipitation tests carried out at pH 8.5 achieve removal yields for all metals > or = 90% using hydrated lime and > or = 83% using caustic soda. Selective precipitation tests alone at pH 5.0 show removal yields > or = 97% for Cr and between 75 and 87% for Al and Pb. Moreover, assays carried out using a stoechiometric addition of Na2S have allowed the separation of Cd (> or = 99%) and Zn (> or = 71%) as metal sulphides (CdS and ZnS). From an economical point of view, the most interesting option seems to be the leachates neutralization at pH 7.0 using Ca(OH)2 combined with the reuse of the treated leachates in the fly ash leaching steps. Metal precipitation cost at pH 7.0 has been estimated to be 22.7 CAN dollars tct-1 using Ca(OH)2, and 26.7 CAN dollars tct-1 using NaOH.

Carbon↗

PubMed1980

Artifactual reduction of high-density lipoprotein cholesterol estimates after dextran sulfate--Mg2+ precipitation.

Estimates of serum or plasma high-density lipoprotein (HDL) concentrations are usually accomplished by measuring the residual cholesterol after polyanionic precipitation of the very low and low-density lipoproteins. A comparison was made of the HDL cholesterol values obtained using both heparin--MnCl2 and dextran sulfate--Mg2+ as precipitating agents. Enzymatic cholesterol determinations showed that the dextran sulfate--Mg2+ method led to gross underestimation of the serum HDL cholesterol concentration (approximately -25%), a difference that was even greater when ultracentrifugally isolated HDL concentrations were assayed rather than serum. Immunochemical determinations of the A-I and B apolipoproteins demonstrated that the discrepancy was attributable neither to incomplete precipitation of the lower density lipoproteins by heparin--MnCl2 nor to precipitation of HDL by dextran sulfate--Mg2+. Incubation of sera with reagents at 37 C, however, minimized or eliminated the differences in results. These findings indicate that temperature is a critical factor when enzymatic quantification of HDL cholesterol is performed after dextran sulfate--Mg2+ precipitation.

Chemical Precipitation↗

PubMed1982

HDL cholesterol quantitation by phosphotungstate-Mg2+ and by dextran sulfate-Mn2+-polyethylene glycol precipitation, both with enzymic cholesterol assay compared with the lipid research method.

Two methods using commercial kits for high density lipoprotein (HDL) cholesterol quantitation were compared with the Lipid Research Clinics (LRC) procedures. HDL cholesterol quantitations on 50 patient specimens by the Lancer HDL cholesterol Rapid Stat Kit (Lancer) with phosphotungstate-Mg2+ precipitation and enzymic cholesterol assay averaged 424 mg/L, and by a method with dextran sulfate-Mn2+-polyethylene glycol (dextran sulfate) precipitation and enzymic cholesterol assay averaged 474 mg/L. By comparison, the LRC method (heparin-Mn2+ precipitation combined with a Liebermann-Burchard reagent cholesterol assay) averaged 478 mg/L. Supernates obtained by the three precipitation methods had similar cholesterol values when analyzed by the LRC assay, suggesting that the observed differences were primarily due to differences between the cholesterol assays. Results were consistent with underestimation by the enzymic assay of cholesterol in the supernates, offset by a positive interference of Mn2+ in the dextran sulfate-produced supernates. Among-day CVs of 4-5% were observed for the Lancer method, and 6-7% for the dextran sulfate method. Sedimentation of precipitates in hypertriglyceridemic specimens was excellent by both methods.

Chemical Precipitation↗

PubMed1987

Modified glycine precipitation technique for the preparation of factor VIII concentrate of high purity and high stability.

A modification of the glycine precipitation method for the production of factor VIII concentrate is reported. In this procedure, cryoprecipitate was prepared, washed and dissolved in buffer. Contaminating fibrinogen was precipitated by the addition of 2.8 M glycine buffer at 30 degrees C. The fibrinogen precipitate was removed by filtration using a layer of glass beads. Factor VIII was then precipitated from the filtrate by adding solid NaCl. After centrifugation, the precipitate was resuspended in buffer, desalted, sterilized by filtration, lyophilized and subjected to dry-heat treatment. The procedure gave a good recovery of factor VIII (280 IU/l) and seems applicable to large-scale production. The resulting factor VIII concentrate was especially characterized by its high purity (10 IU/mg) and stability both in the liquid state and during dry-heat treatment.

Chemical Precipitation↗

PubMed1966

Precipitate formation in the strontium-phosphate system.

Study of the precipitation process in the aqueous Sr(OH)(2)-H(3)PO(4) system, in order to elucidate the phase transformations and the nature of the final solid phases, shows that over much of the range of compositions studied the initial precipitate is poorly crystalline; the x-ray pattern resembles that of strontium hydroxyapatite but has a strontium: phosphorus molar ratio close to 1.3. Within 1 hour the initial precipitate changes to a stable crystalline phase (or phases), with corresponding change, either up or down, in the strontium: phosphorus ratio. At high ratios of Sr(OH)(2) to H(2)PO(4) the initial precipitate is Sr(3)(PO(4))(2)-4H(2)O, which then converts to a phase having the x-ray diffraction pattern of strontium hydroxyapatite, but having a strontium: phosphorus ratio that depends somewhat on the initial ratio of Sr(OH)(2) to H(3)PO(4) used in the precipitation.

Apatites↗

PubMed1985

Effect of cimetidine on enzyme inactivation, bile acid precipitation, and lipid solubilisation in pancreatic steatorrhoea due to cystic fibrosis.

In pancreatic steatorrhoea, both pH-dependent bile acid precipitation and enzyme inactivation may limit the efficacy of pancreatic enzyme supplements and both may be preventable by addition of cimetidine. To separate these effects we compared postprandial jejunal aspirate from eight adults with steatorrhoea due to cystic fibrosis on three randomised treatment regimens (pancreatin, cimetidine, and both together). We also compared the results with those of previous studies of patients on no treatment, and of healthy subjects. On pancreatin 60% of the test meal entered the jejunum at pH less than 5 compared with 17% in health. Lipase concentration and lipolysis increased over the values on no treatment (14.2 vs 4.4 U/l, p less than 0.01; 16% vs 11%, p less than 0.02) but bile acid precipitation was not reduced (38% vs 27%, NS), and aqueous-phase lipid concentration decreased (6.7 vs 8.6 mM/l, p less than 0.05). On cimetidine, bile acid precipitation fell (19% vs 38%, p less than 0.05); although lipase concentration and lipolysis were lower than on pancreatin (4.8 U/l vs 14.2 U/l, p less than 0.01; 9% vs 16%, p less than 0.01) lipid solubilisation increased (8.8 vs 6.7 mM/l, p less than 0.05). On the combination, there was a marked improvement (p less than 0.02) in lipid solubilisation (18.3 mM/l), reflecting the improvement both in lipase (38.4 U/l) and lipolysis (24%), and in bile acid precipitation (5.6%). We conclude that the efficacy of pancreatin is limited by pH-dependent bile acid precipitation in addition to enzyme inactivation. The action of cimetidine in improving the efficacy of pancreatin depends on prevention of both these effects.

Adolescent↗

PubMed1986

Contrast medium precipitation during abdominal CT.

Abdominal computed tomography (CT) scans of 55 patients who had ingested Gastrografin (meglumine diatrizoate and diatrizoate sodium) diluted to 2% with tap water and flavored with a commercial fruit juice base were reviewed. Twenty patients (36%) demonstrated intraluminal precipitation of Gastrografin shown by focal areas of markedly increased attenuation within the gastric lumen or trapped within gastric folds. Beam-hardening artifact produced by precipitation was observed, which limited the diagnostic value of some examinations. In vitro CT scans of the same Gastrografin solution titrated with hydrochloric acid or sodium hydroxide showed that by raising the pH of the solution, precipitation was virtually eliminated. Fifty-one CT scans of the abdomen using a buffered Gastrografin solution demonstrated precipitation in only five patients. Properly buffered dilute oral Gastrografin solutions should significantly decrease the prevalence of precipitation during abdominal CT examinations.

Beverages↗

PubMed1999

A study of struvite precipitation and urease activity in bacteria isolated from patients with urinary infections and their possible involvement in the formation of renal calculi.

Struvite precipitation and urease activity were studied in 72 bacterial strains isolated from patients with an urinary infection. The results revealed struvite precipitation by urease-positive, urease-negative and weakly urealitic bacteria. However, in some strains of each of these groups, no precipitation was observed. Variations in pH within the cultures were also investigated as were the mineralogy and morphology of the crystals precipitated using X-ray diffraction and scanning electron microscopy, respectively. The role of urease activity and alkalinization of the medium in struvite precipitation is discussed, as is the possible involvement of bacteria in nucleation processes. We concluded that urease-negative bacteria or those with weak urease activity may also be involved in the formation of struvite renal calculi although urease-positive bacteria seem to make a greater contribution.

Chemical Precipitation↗

PubMed1989

The effect of silicic acid on calcium phosphate precipitation.

So that a possible involvement in the mineralization of dental plaque could be investigated, the effects of silicic acid on calcium phosphate precipitation were assessed in vitro. By measuring the decrease in Ca2+ concentration (by means of ion-selective electrodes), we determined both spontaneous precipitation and seeded crystal growth from solutions that contained 1 mmol/L calcium, 7.5 mmol/L phosphate, 50 mmol/L Hepes pH 7.2, and various amounts of silicic acid. Polymerized silicic acid, but not its monomer, was found both to cause a 60% reduction in the lag period that precedes spontaneous precipitation and to enhance the growth rate of seeded hydroxyapatite crystals. Silica suspensions showed effects similar to those of polysilicic acid. In all cases, the precipitated material was found to be hydroxyapatite. Whereas seeded brushite crystals grew slowly without silicic acid, hydroxyapatite was the only mineral detected after crystal growth in the presence of silicic acid. Apparently, polysilicic acid acted as a substrate for hydroxyapatite nucleation, inducing secondary nuclei on both hydroxyapatite and brushite crystals. The finding that polysilicic acid could overcome part of the inhibitory effect of a phosphoprotein on calcium phosphate precipitation gave additional support for the idea that polysilicic acid and silica may promote the formation of dental calculus.

Calcium Phosphates↗

PubMed1991

Induction of calcium phosphate precipitation by titanium dioxide.

Titanium powder and various titanium dioxides were tested for their capacity to reduce the induction time for calcium phosphate precipitation from supersaturated solutions. Only after a pretreatment aimed at increasing its oxide surface layer was titanium powder found to accelerate the precipitation from solutions containing 2 mmol/L CaCl2, 2 mmol/L KH2PO4, 50 mmol/L Hepes, pH 7.2, and to induce precipitation from metastable solutions containing 1.2 mmol/L CaCl2, 1.2 mmol/L KH2PO4, 50 mmol/L Hepes, pH 7.2, at 37 degrees C. Even stronger effects were found when suspensions of the titanium dioxides anatase or rutile (10-50 micrograms/mL) were added to these solutions. TiO2 appeared to serve as a reactive substrate for secondary nucleation at a wide range of calcium-to-phosphate ratios and concentrations, even in the presence of 40 mg/mL bovine serum albumin, which completely inhibited precipitation in control incubations. These results suggest that the oxide surface layer of titanium implants may induce calcium phosphate precipitation in the metal-to-bone interface, which may play a role in the integration of such implants in bone.

Calcium Phosphates↗

PubMed2004

Predicting the precipitation of poorly soluble weak bases upon entry in the small intestine.

Solubility and dissolution relationships in the gastrointestinal tract can be critical for the oral bioavailability of poorly soluble drugs. In the case of poorly soluble weak bases, the possibility of drug precipitation upon entry into the small intestine may also affect the amount of drug available for uptake through the intestinal mucosa. To simulate the transfer out of the stomach into the intestine, a transfer model was devised, in which a solution of the drug in simulated gastric fluid is continuously pumped into a simulated intestinal fluid, and drug precipitation in the acceptor medium is examined via concentration-time measurements. The in-vitro precipitation of three poorly soluble weakly basic drugs, dipyridamole, BIBU 104 XX and BIMT 17 BS, was investigated. For all three, extensive supersaturation was achieved in the acceptor medium. Under simulated fasted-state conditions, precipitation occurred for all three compounds whereas under simulated fed-state conditions, the higher concentrations of bile components and the lower pH value in the acceptor medium inhibited precipitation at concentrations corresponding to usual doses in all cases. Comparison with pharmacokinetic data indicated that a combination of transfer model data with solubility and dissolution profiles should lead to better predictions of in-vivo behaviour of poorly soluble weak bases.

Benzimidazoles↗

PubMed1988

[Clinical studies on abnormal thyroid stimulators in patients with Graves' disease. I. A sensitive assay for thyroid-stimulating antibodies using cultured porcine thyroid cells and polyethylene glycol precipitation of serum].

The activities of thyroid-stimulating antibody (TSAb) in serum from patients with Graves' disease were measured by a sensitive assay, using cultured porcine thyroid cells and the precipitation from serum with polyethylene glycol (PEG), and the activities were compared with those of thyrotropin binding inhibitor immunoglobulin (TBII), measured by the commercial assay kit. Porcine thyroid cells after digestion were cultured for 15-18 hours with TSH of 1-10,000 microU/ml or the precipitations of sera from normal subjects and patients with Graves' disease or Hashimoto's thyroiditis, and then the cAMP levels in the culture medium were determined by the commercial RIA assay kit (Yamasa). The precipitation was obtained by adding 0.5 ml of 30% PEG solution to 0.5 ml serum, and was resuspended with 0.6 ml of Hanks' medium without NaCl, containing 1.5% bovine serum albumin, 20mM Hepes and 0.5 mM 3-isobutyl-1-methylxanthine. The precipitation contained about 85% of immunoglobulin and 63% of albumin of the original amount of the serum, as well as substantial TSH, when the original serum contained TSH more than 40 microU/ml. When the PEG precipitations from 10 normal subjects were incubated with the thyroid cells of 4 X 10(5) cells, the cAMP releases into the medium ranged from 83 to 124%, when the mean value was calculated as 100%. Therefore, the cAMP release of more than 130% of the amount released into the culture medium incubated with normal IgG was judged as positive TSAb activity. The minimum detectable quantities were regarded as about 5 microU/ml TSH equivalent. TSAb and TBII activities were detected in 48 (92%) and 50 (96%) of 52 patients with untreated hyperthyroid Graves' disease, respectively, and either TSAb or TBII activities were detected in 16 (80%) of 20 patients with Graves' disease maintained in a clinically euthyroid state by treatment with antithyroid drugs. TBII was positive in 10(50%) of these patients. Some patients showed distinct discrepancies in these two activities, although there was a significant positive correlation between TSAb and TBII activities (r = 0.53, p less than 0.01) in patients with untreated Graves' disease. In these patients, TSAb activities showed a significant positive correlation with values for 99mTc thyroid uptake, determined 30 min after the injection. However, they did not show any significant correlation with serum T4 or T3 concentrations. Similarly, TBII showed significant correlations with goiter size and 99mTc thyroid uptake. To conclude, the present assay for TSAb is sensitive and reproducible.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1995

Effect of Tamm-Horsfall protein on calcium oxalate precipitation.

The effect of Tamm-Horsfall protein isolated from urine of healthy subjects on calcium oxalate precipitation was studied in model systems of precipitation. The study was performed using following conditions: concentrations of calcium chloride 10 mmol/l, sodium chloride 150 mmol/l, oxalic acid 300 mumol/l; pH 6.0, and temperature 310 K. The concentration of Tamm-Horsfall protein varied between 1-10 mg/l. The kinetics of calcium oxalate precipitation was observed by measuring the number and volume of particles in the suspension, and the precipitate composition by an optic microscope. In all the studied systems, the precipitate morphology corresponded to pure calcium oxalate monohydrate. Tamm-Horsfall protein was found to inhibit the growth of calcium oxalate monohydrate crystals and stimulate their aggregation in the given experimental conditions. Both effects were enhanced by increase in the concentrations of Tamm-Horsfall protein and were most pronounced at the concentration of Tamm-Horsfall protein of 10 mg/l.

Adult↗