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Evaluation of precipitation and direct methods for HDL-cholesterol assay by HPLC.

HDL-cholesterol (HDL-C) values measured by precipitation (sodium phosphotungstate-MgCl2) and direct methods were compared with those obtained by HPLC with a new column (TSKgel Lipopropak) and an eluent (TSKeluent LP-1). The HDL-C values determined by the precipitation method were significantly (P <0.001) lower than those by the HPLC method, whereas the HDL-C values by the direct method were slightly but significantly higher (P <0.02) than those by the HPLC method. A quantitative HPLC analysis of the cholesterol concentration in HDL and non-HDL fractions in the supernatant of serum separated by precipitation reagents with different MgCl2 concentrations ranging from 7.3 to 44 mmol/L revealed that reagents with >22 mmol/L MgCl2 precipitated part of HDL as well as non-HDL lipoproteins. The HPLC method providing quantitative and qualitative information with high precision was regarded as being a reliable approach for HDL-C assay. The HPLC can be also used to evaluate alternative methods for cholesterol assay.

Chemical Precipitation↗

Determination of monoethylglycinexylidide by fluorescence polarization immunoassay in highly icteric serum samples: modified precipitation procedure and HPLC compared.

Hyperbilirubinemia, which frequently occurs in severe liver disease, interferes with the fluorescence polarization immunoassay (FPIA) monoethylglycinexylidide (MEGX) assay manufactured by Abbott Diagnostics. Because the MEGX test is particularly helpful in this clinical situation, strategies have been developed to overcome this problem. Precipitation of serum with the Abbott Digoxin II precipitation reagent eliminates bilirubin. Therefore, we compared FPIA results after precipitation of 81 icteric samples from 27 MEGX tests to results obtained using a validated HPLC method. The precipitation did not substantially alter the performance characteristics of FPIA: detection limit, 8 microg/L; between-days imprecision, 5.3-6.2%; recovery, 102-104% (50-200 microg/L). This pretreatment of serum did not eliminate all interference, and only a poor correlation was observed between serum MEGX concentrations measured with HPLC or modified FPIA (r2 = 0.46; S(y/x) = 20.0 microg/L). In contrast, MEGX formation values calculated by subtraction of the prelidocaine MEGX concentration were in close agreement (r2 = 0.98; S(y/x) = 2.3 microg/L). Because only MEGX formation is clinically relevant, this modified FPIA procedure offers a simple and rapid alternative to HPLC.

Chemical Precipitation↗

Nanostructured pure gamma-Fe2O3 via forced precipitation in an organic solvent.

Pure maghemite, gamma-Fe(2)O(3), was prepared as ultra fine particles in the nanometer-sized range via the forced precipitation method in an organic solvent. The precipitation of iron(III) ions, from iron(III) chloride in 2-propanol led selectively to highly dispersed particles of ferrihydrite, which upon treatment with temperatures higher than 200 degrees C under dynamic vacuum resulted in high-surface-area particles of gamma-Fe(2)O(3). Precipitation in water also led to ferrihydrite, but the final product, after heating at 300 degrees C, contained a mixture of gamma-Fe(2)O(3) and alpha-Fe(2)O(3) (hematite). The precipitation from iron(III) nitrate in water resulted in goethite which was converted to hematite upon heating. On the other hand, the final product in 2-propanol was a mixture of maghemite and hematite. The products were characterized by FTIR, TGA, XRD, and gas sorption analysis. Nitrogen gas adsorption studies for the pure gamma-Fe(2)O(3) samples revealed mesoporous particles with high surface areas in the range of 70-120 m(2) g(-1) after heat treatment at 300 degrees C. The gamma-Fe(2)O(3) particles retained their gamma-phase as well as their mesoporous structure at relatively high temperatures, as high as 400 degrees C.

Journal Article↗

Treatment of hypercholesterolemia by precipitation of lipoproteins with dextran sulfate.

An on-line continuous system for the selective precipitation of low-density lipoproteins (LDL) and very low-density lipoproteins (VLDL) has been devised and tested. This system conserves high-density lipoproteins (HDL) and other plasma macromolecules. LDL and VLDL are precipitated from plasma using 10-35 mg/dl dextran sulfate (Mr 5,000) in the presence of 55 mM calcium with a reduced concentration of monovalent cations. The plasma is obtained by membrane filtration of whole blood using the COBE Centry TPE System (Cobe Laboratories Inc, Lakewood, Co.). The precipitated LDL plus VLDL is removed by filtration, and the electrolytes are restored by dialysis. The plasma minus LDL plus VLDL is then returned to the patient. Four patients with heterozygous familial hypercholesterolemia (type II) were treated 70 times. The mean pretreatment serum cholesterol was 383 mg/dl. The mean reductions in plasma components were: LDL plus VLDL 63%; HDL 27%; fibrinogen 19%; albumin 15%; IgG 20%; IgA 19%; IgM 25%; C3 30%; and C4 27%. The cholesterol returned to near normal values in approximately 2 weeks after each treatment. Four normal volunteers were each treated one time. These individuals had a mean pretreatment serum cholesterol of 201 mg/dl. The mean reduction in plasma components were: LDL plus VLDL 70%; HDL 27%; fibrinogen 24%; albumin 14%; IgG 18%; IgA 17%; IgM 20%; C3 27%; C4 22%; C3 proactivator 12%; alpha 1-antitrypsin 17%; ceruloplasma 17%; transferrin 18%; alpha 2-macroglobulin 17%; and orosomucoid 13%. It is our conclusion that dextran sulfate precipitation is an effective on-line means of selectively removing LDL plus VLDL from plasma while conserving HDL and other plasma macromolecules.(ABSTRACT TRUNCATED AT 250 WORDS)

Algorithms↗

Techniques for converting Golgi precipitate in CNS neurons into stable electron microscopic markers.

Direct electron microscopy of nervous tissue stained with the Golgi impregnation method is unsatisfactory because the cytoplasm of the cell bodies and processes of the impregnated neurons are completely filled with a compact precipitate of electron dense silver chromate. This precipitate entirely obscures the cytological details of the impregnated neurons. Because of its solidity and instability in aqueous solutions, the silver chromate is also a source of inconvenience during the preparation of the ultrathin sections. This review summarizes methods that have been developed with the aim of replacing the Golgi precipitate in CNS neurons with a more convenient electron dense material--for example, heavy metal salts or metallic particles. Conversion of the precipitate into a stable electron dense marker is done before the material is embedded for electron microscopy. The methods include lead, gold, and bromide substitution, treatment with ammonia, direct chemical reduction into metallic silver, and photoreduction of the silver chromate into silver through irradiation with ultraviolet light.

Aminophenols↗

Interaction of Citric or Hydrochloric Acid with Calcium Fluorapatite: Precipitation of Calcium Fluoride.

The interaction of citric acid (H(3)Ci) with calcium fluorapatite(Ca(10) F(2)(PO(4))(6)) was explored for two reasons: (i) to determine the role of the acid in the dissolution process and hence in the mechanism of tooth fluoridation and (ii) to determine whether there is any formation of calcium citrate. It was found that the concentration of calcium or fluoride ions is not stoichiometric with respect to that of phosphate ions in the solution and the stoichiometric deficiency of the amount of fluoride ions in the solution is twice that of the calcium ions and this demonstrates that some calcium fluoride is precipitated. The interaction may be represented by the following chemical equation (pH approximately 2.5): where x (presently < 1) is related to the amount of CaF(2) that has precipitated and may be calculated from the experimental ratio of Ca to P in the solution. In order to establish that the interaction occurs generally with all acids, the reaction of hydrochloric acid with fluoroapatite was studied, and exactly analogous behavior was observed. These facts are also in accord with the solubility of fluorapatite and calcium fluoride. When the amounts of Ca and P in solution are corrected for the precipitated CaF(2), the ratio of Ca to P becomes stoichiometric (= 1.67). Preliminary X-ray analysis of the reacted fluorapatite showed that it contained calcium fluoride. The precipitated CaF(2) may act as a reservoir for the subsequent fluoridation of the mineral and may inhibit bacterial action in the mouth. The concentration of citrate ions does not change in solution, and no formation of calcium citrate is observed.

Journal Article↗

Sandbed Consolidation with Mineral Precipitation.

A new method has been developed to prevent sand reentrainment during oil production from unconsolidated or poorly consolidated reservoir formations. Consolidation of the zformation around the well is achieved through in situ precipitation of a sparingly soluble salt, namely, calcium phosphate. Control of the depth of salt formation is achieved by alternating injection, mixing, and reaction of two aqueous solutions of calcium chloride and potassium phosphate. Calcium phosphate crystals precipitate and grow on the grain surfaces, forming sufficiently uniform coatings. The formation of relatively uniform coatings on the grains causes an acceptably small decrease of the permeability, which is a feature of primary importance for oil production. The grains are gradually "cemented" with bridges of calcium phosphate crystallites and form a consolidated and still porous structure. As a result, the rate of hydrocarbon production for the problematic reservoir can be increased considerably without undesirable reentrainment of sand. The proposed method for consolidation has been successfully tested in sandbeds. Several series of experiments have been carried out under diverse conditions to establish the optimum parameter values for the implementation of this method. A set of optimum conditions at 25 degrees C were determined and these conditions gave satisfactory consolidation with permeability loss of ca. 60% of the initial value. The conditions of precipitation were chosen so that the precipitated phase was octacalcium phosphate [Ca(4)H(PO(4))(3).2.5H(2)O], along with its byproduct hydroxyapatite [Ca(5)(PO(4))(3)OH]. Experiments were also carried out at 70 degrees C and have shown that it is feasible to consolidate loose sandpacks at oil reservoir conditions. Copyright 2000 Academic Press.

Journal Article↗

In Situ Monitoring of Generation and Precipitation of Ferric Hydroxide Sol with a Piezoelectric Quartz Crystal Impedance Analyzer.

The piezoelectric quartz crystal (PQC) impedance technique was applied to monitor in situ generation and precipitation of the ferric hydroxide sol in aqueous solutions at 90 degrees C. Equivalent circuit parameters and resonant frequencies as well as the half-peak width of the electroacoustic conductance spectrum deltaf(G1/2) for the PQC resonance were obtained and analyzed. Three stages, sol generation and simultaneous adsorption, adsorption equilibrium, and precipitation of ferric hydroxide sol could be identified in the process of adding the ferric nitrate solution into the hot water. A scheme of two consecutive reactions occurring at the electrode/solution interface was used to analyze the adsorption kinetics of ferric hydroxide sol onto the Au electrode. In addition, the electrolyte-induced precipitation of the colloid was monitored and discussed. Temperature effect on the PQC resonance behavior in liquid was also investigated. Since the PQC impedance technique provides multidimensional piezoelectric information in situ, it is highly recommended for studying the process of sol-gel generation and precipitation. Copyright 2001 Academic Press.

Journal Article↗

Multicolour preparations for in situ hybridization using precipitating enzyme cytochemistry in combination with reflection contrast microscopy.

We have further developed a method for the detection of different enzyme cytochemical reaction products by means of reflection contrast microscopy (RCM). By embedding these enzyme precipitates in a protein matrix, we were able to prevent the reaction products from dissolving in immersion oil, which is required for RCM analysis. The applicability of the RCM procedure is, therefore, extended to a range of cytochemical enzyme precipitation methods, which normally result in oil soluble reaction products. To test their usefulness, these enzyme precipitates have been used in single- and well as double-label in situ hybridization (ISH) procedures to visualize a number of DNA target sequences by several different reflection colours, i.e. white, yellow and red. Three repetitive DNA probes for the (sub)centromeric regions of chromosomes 1, 7 and 17, as well as a repetitive DNA probe for the telomeric region of chromosome 1, and two cosmid DNA probes (40 kb each) for both arms of chromosome 11 could be detected with high efficiency in both interphase and metaphase preparations. Moreover the enzyme precipitates were shown to be stable upon exposure to excitation light or upon storage. It may be concluded that these findings render RCM a sensitive method for the visualization of multiple targets in biological specimens.

Carcinoma, Transitional Cell↗

Sodium and calcium localization in cells and tissues by precipitation with antimonate: a quantitative study.

Komnick's antimonate technique, which was devised to localize Na+ in cells and tissues, was studied quantitatively. Some modifications, as well as its application to Ca2+ localization, were also investigated. We combined measurements of Na+ and Ca2+ retention in plant roots during the various procedures, electron microscopy, autoradiography, and semiquantitative X-ray microanalysis. We were able to show that (at least in barley roots) antimonate does not precipitate at all with Na+, irrespective of the Na+ content of the tissue or the method of antimonate application. (Even during precipitative freeze dissolution or after freeze drying, no Na+ is precipitated.) By means of Komnick's antimonate technique Ca2+ is trapped within the tissue, but only after serious dislocation. Perspectives for reliable localization of diffusible ions in cells and tissues, by precipitation simultaneously with conventional fixations, are bad.

Antimony↗

Binding and precipitating activities of Erythrina lectins with complex type carbohydrates and synthetic cluster glycosides. A comparative study of the lectins from E. corallodendron, E. cristagalli, E. flabelliformis, and E. indica.

Erythrina lectins possess similar structural and carbohydrate binding properties. Recently, tri- and tetra-antennary complex type carbohydrates with non-reducing terminal galactose residues have been shown to be precipitated as tri- and tetravalent ligands, respectively, with certain Erythrina lectins [Bhattacharyya L, Haraldsson M, Brewer CF (1988) Biochemistry 27:1034-41]. The present work describes a comparative study of the binding and precipitating activities of four Erythrina lectins, viz., E. corallodendron, E. cristagalli, E. flabelliformis, and E. indica, with multi-antennary complex type carbohydrates and synthetic cluster glycosides. The results show that though their binding affinities are very similar, the Erythrina lectins show large differences in their precipitating activities with the carbohydrates. The results also indicate significant dependence of the precipitating activities of the lectins on the core structure of the carbohydrates. These findings provide a new dimension to the structure-activity relationship of the lectins and their interactions with asparagine-linked carbohydrates.

Carbohydrate Conformation↗

Precipitated withdrawal in squirrel monkeys after repeated daily oral administration of alprazolam, diazepam, flunitrazepam or oxazepam.

The lowest dose of alprazolam, diazepam, flunitrazepam and oxazepam consistently to induce loss of righting reflex in squirrel monkeys or vehicle was orally administered to monkeys on 18 consecutive days: 2 mg/kg alprazolam (n = 4), 30 mg/kg diazepam (n = 4), 1 mg/kg flunitrazepam (n = 4), 280 mg/kg oxazepam (n = 5), or vehicle (n = 4). Tolerance developed rapidly for loss of righting reflex, more slowly for sleep and only minimally for muscle relaxation observed during the period immediately following daily oral administration. Injection of the specific benzodiazepine receptor antagonist flumazenil (10 mg/kg i.v.) 5 h after the ninth daily oral treatment produced signs of precipitated withdrawal (tremor, vomiting and/or convulsions) in one alprazolam-, four diazepam-, one flunitrazepam- and four oxazepam-treated monkeys, but not in the vehicle-treated monkeys. Physiological saline injected intravenously several days later under these same experimental conditions failed to provoke a precipitated withdrawal reaction. When flumazenil-induced precipitated withdrawal was again evaluated after the 18th daily oral treatment, withdrawal signs were observed in all alprazolam- and all diazepam-treated monkeys, as well as in three flunitrazepam- and three oxazepam-treated monkeys, but not in the vehicle-treated monkeys (convulsions were observed in one alprazolam-, two diazepam-, one flunitrazepam- and two oxazepam-treated monkeys). No signs of spontaneous withdrawal were observed in any of the monkeys during a subsequent 3-week drug-free period. Thus, repeated administration of approximately equieffective doses of these four benzodiazepines resulted in a similar development of tolerance and physical dependence (indicated by the occurrence of a precipitated withdrawal reaction).

Administration, Oral↗

Clonidine infusions into the locus coeruleus attenuate behavioral and neurochemical changes associated with naloxone-precipitated withdrawal.

Clonidine, an alpha-2-adrenergic agonist, suppresses signs of opiate withdrawal in animals and in man. Electrical or chemical stimulation of the nucleus locus coeruleus (LC) increases noradrenergic activity and brain concentration of the noradrenergic metabolite MHPG, and produces many signs of opiate withdrawal. Thus, clonidine's ability to attenuate withdrawal might be due to the reduction of noradrenergic neuronal activity originating in the LC, but additional alpha-2-adrenergic receptors throughout the body and other mechanisms may also play a role. The present study explored the neuroanatomical and pharmacological selectivity of alpha-2-adrenergic receptors of the LC in the anti-withdrawal action of clonidine. Experiment 1 tested the hypothesis that behavioral and biochemical measures of naloxone-precipitated withdrawal from morphine would be blocked by infusions of clonidine (0.6 or 2.4 micrograms/microliters) into the LC. Significant reductions were observed in the occurrence of diarrhea, ptosis, weight loss and wet-dog shakes. Clonidine also reversed the naloxone-precipitated increase in hippocampus MHPG concentration. In experiment 2 subjects received an LC infusion or IP injection of a non-lipophilic alpha-2-agonist (ST-91), which does not penetrate the blood-brain barrier, or of clonidine into the dorsal parabrachial nucleus (DPB) to test the selectivity of the effects of clonidine infusions into the LC. ST-91 infusions into the LC reduced several of the observed withdrawal signs and increased others (e.g., jumping). Although peripheral injections of ST-91 attenuated some of the checked signs associated with naloxone-precipitated withdrawal, the frequency of wet-dog shakes was not reduced. ST-91 infusions into the LC, but not systemic ST-91 administration, prevented the withdrawal-induced increase in hippocampus MHPG concentration. Clonidine infused lateral to the LC into the DPB did not significantly attenuate withdrawal or reduce hippocampus MHPG levels. These results provide behavioral and biochemical evidence to support the suggestion that clonidine significantly attenuates naloxone-precipitated withdrawal through an interaction with noradrenergic neurons located in the vicinity of the LC.

Animals↗

A comparison of the effects of clonidine and CNQX infusion into the locus coeruleus and the amygdala on naloxone-precipitated opiate withdrawal in the rat.

Both the locus coeruleus (LC) and the amygdala have been implicated in aspects of opiate dependence and withdrawal. The LC is known to be one of the most sensitive sites for precipitating withdrawal behaviors after local opiate antagonist infusions in morphine-dependent subjects. The amygdala is also known to mediate antagonist-induced withdrawal behaviors and aversive motivational states. The goal of the present study was to evaluate directly the ability of noradrenergic agonists and glutamatergic antagonists to attenuate naloxone-precipitated withdrawal behaviors when infused into the LC or the central nucleus of the amygdala (CeA). The alpha-2-noradrenergic agonists clonidine or ST-91 were infused into the CeA to compare the effects of noradrenergic activation in the CeA to the attenuation of withdrawal previously observed in rats infused with clonidine into the LC, since the LC and CeA are known to contain co-localized opiate and noradrenergic receptors. The effects of microinfusions of the non-NMDA excitatory amino acid antagonist 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX) were also infused into the LC and CeA since opiate withdrawal is associated with increased glutamatergic transmission. Intra-CeA clonidine or ST-91 (2.4 microg/0.5 microl or 1.0 microl) produced significant reductions primarily in the occurrence of irritability. Conversely, intra-CeA or intra-LC infusions of CNQX (2.5 microg/0.5 microl) significantly attenuated naloxone-precipitated withdrawal, an effect similar to the attenuation previously observed after intra-LC clonidine infusions. These data demonstrate the specific behavioral effects of altering glutamatergic and noradrenergic neurotransmission in the LC or CeA during naloxone-precipitated opiate withdrawal. Elucidation of the neuroanatomical circuitry involved in opiate withdrawal should increase our understanding of the neuroadaptations associated with drug dependence and subsequent withdrawal behavior.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Response of net ecosystem gas exchange to a simulated precipitation pulse in a semi-arid grassland: the role of native versus non-native grasses and soil texture.

Physiological activity and structural dynamics in arid and semi-arid ecosystems are driven by discrete inputs or "pulses" of growing season precipitation. Here we describe the short-term dynamics of ecosystem physiology in experimental stands of native (Heteropogon contortus) and invasive (Eragrostis lehmanniana) grasses to an irrigation pulse across two geomorphic surfaces with distinctly different soils: a Pleistocene-aged surface with high clay content in a strongly horizonated soil, and a Holocene-aged surface with low clay content in homogenously structured soils. We evaluated whole-ecosystem and leaf-level CO2 and H2O exchange, soil CO2 efflux, along with plant and soil water status to understand potential constraints on whole-ecosystem carbon exchange during the initiation of the summer monsoon season. Prior to the irrigation pulse, both invasive and native grasses had less negative pre-dawn water potentials (Psipd), greater leaf photosynthetic rates (Anet) and stomatal conductance (gs), and greater rates of net ecosystem carbon exchange (NEE) on the Pleistocene surface than on the Holocene. Twenty-four hours following the experimental application of a 39 mm irrigation pulse, soil CO2 efflux increased leading to all plots losing CO2 to the atmosphere over the course of a day. Invasive species stands had greater evapotranspiration rates (ET) immediately following the precipitation pulse than did native stands, while maximum instantaneous NEE increased for both species and surfaces at roughly the same rate. The differential ET patterns through time were correlated with an earlier decline in NEE in the invasive species as compared to the native species plots. Plots with invasive species accumulated between 5% and 33% of the carbon that plots with the native species accumulated over the 15-day pulse period. Taken together, these results indicate that system CO2 efflux (both the physical displacement of soil CO2 by water along with plant and microbial respiration) strongly controls whole-ecosystem carbon exchange during precipitation pulses. Since CO2 and H2O loss to the atmosphere was partially driven by species effects on soil microclimate, understanding the mechanistic relationships between the soil characteristics, plant ecophysiological responses, and canopy structural dynamics will be important for understanding the effects of shifting precipitation and vegetation patterns in semi-arid environments.

Analysis of Variance↗

Precipitants of elderly psychiatric patient assaults on staff: preliminary empirical inquiry.

Although there have been several studies of the characteristics of psychiatric patient assailants, there have been only six comprehensive, empirical assessments of precipitants to these assaults and no precipitant study has focused solely on elderly psychiatric patient assailants. This one and one-half year, retrospective study continued the inquiry into the nature of patient assault precipitants and focused only on elderly assailants. Older, male patients with a diagnosis of schizophrenia and histories of violence toward others and substance use disorder physically assaulted primarily male, mental health workers. These staff victims experienced disruptions in the domains of mastery, attachment, and meaning as well as the symptomatology associated with psychological trauma. The most common precipitants to these assaults were denial of services and acute psychosis. The findings and implications for health care providers in long-term care settings where elderly psychiatric patients reside are discussed.

Aged↗

Specific precipitating antibody to human urinary erythropoietin.

In the present experiments, human urinary erythropoietin was purified from the urine of a patient suffering from pure red cell aplasia, using ethanol precipitation, gel filtration on the Sephadex G-100 column and electrofocusing. The anti-human urinary erythropoietin was prepared by injecting the isolated erythropoietin with Freund's complete adjuvant into rabbits intracutaneously. The specificity of the anti-human urinary erythropoietin was ascertained by a double diffusion technique and immunoelectrophoresis, observing a single precipitation line at the alpha2-globulin region against human serum or human urinary erythropoietin (Standard B). This precipitating anti-human urinary erythropoietin inhibited completely the effects of erythropoietin (Standard B) on the heme synthesis of bone marrow cells in vitro. The precipitating antigen with the specific anti-human urinary erythropoietin, that is, erythropoietic stimulating factor of the fraction 2, had the ability to enhance heme synthesis in bone marrow cells in vitro. The specific anti-human urinary erythropoietin makes it possible to determine the serum erythropoietin levels in various diseases by means of quantitative immunoelectrophoresis.

Animals↗

Interference of blood clotting factors in the determination of HDL2- and HDL3-cholesterol by the dual precipitation method.

Previously we found that in the dual precipitation method for determination of HDL2- and HDL3-cholesterol the optimal concentration of dextran sulfate was 0.87 g/l instead of 1.3 or 1.5 g/l. In the present study, we performed the dual precipitation method both in serum and in EDTA-plasma samples in order to investigate whether the method is sensitive for plasma-serum differences. It was found, that in the analysis of EDTA-plasma samples a twice higher final concentration of dextran sulfate was needed. Addition of the dextran sulfate resulted in the appearance of fibrin structures in the HDL-fraction which may have diminished the effective dextran sulfate concentration in the precipitation reaction. Although the use of plasma is advised in the analysis of lipoproteins, the use of EDTA-plasma in precipitation methods for determination of HDL-cholesterol and its subfractions cannot be recommended.

Blood Coagulation Factors↗