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Endothelial precipitates and laser flare photometry in patients with acquired immunodeficiency syndrome: a screening test for cytomegalovirus retinitis?

Patients with acquired immunodeficiency syndrome (AIDS) who present with cytomegalovirus (CMV) retinitis show pathognomonic endothelial precipitates suggestive of primary anterior uveitis or secondary changes due to a spill-over from the posterior chamber. Laser flare photometry allows quantification of the intensity of anterior affection. We wanted to establish anterior-chamber flare values in AIDS patients with and without CMV retinitis and to find out whether CMV retinitis is preceded by an elevation of the flare value. In all, 25 men with AIDS who presented with CMV retinitis and 27 who did not have CMV retinitis but showed a CD4 count of < or = 200 cells/microliter blood were enrolled in a prospective study. Slit-lamp examination was performed, followed by indirect ophthalmoscopy and laser flare photometry after dilation of the pupil with tropicamide eye drops. Patients with CMV retinitis were followed every 10 days and the others, every 4 weeks. A group of 51 human immunodeficiency virus (HIV)-negative men served as a control group. AIDS patients with CMV retinitis showed a significantly higher flare count in the affected eye (12.4 photons/ms; n = 26) as compared with the unaffected partner eye (4.2 photons/ms; P < or = 0.0001; n = 18) and with eyes of AIDS patients without CMV retinitis (4.1 photons/ms; P < or = 0.0001; n = 50). The count in the latter eyes was also significantly higher than the control value (3.1 photons/ms; P < or = 0.0001; n = 102). Typical reticulate endothelial precipitates were found in 92% of AIDS patients with CMV retinitis. During the study, five eyes of three patients developed a fresh CMV retinitis, but a preceding rise in the flare count was not observed. Laser flare photometry follows the occurrence of pathognomonic reticulate endothelial precipitates. It lags behind the development and the extension of CMV retinitis. Therefore, it cannot be used as a screening test for early detection of CMV retinitis.

AIDS-Related Opportunistic Infections↗

[A simple method of studying oxidation resistance of heparin-precipitated blood beta lipoproteins].

Decrease of beta-lipoprotein resistance to copper-dependent oxidation is an important diagnostic criterion of risk of atherosclerosis. We developed a simple method for assessing resistance of heparin-precipitated beta-lipoproteins to oxidation induced by 50 microM copper ions. A strong positive correlation between the oxidation of precipitated and ultracentrifuged beta-lipoproteins in the presence of 50 and 10 microM copper ions, respectively, was revealed. In coronary patients oxidation resistance of precipitated beta-lipoproteins was decreased in comparison with that in healthy donors. The method is simple, informative, and easily available for clinical biochemical laboratories.

Adult↗

Acute precipitants of congestive heart failure exacerbations.

BACKGROUND: Few studies have prospectively and systematically explored the factors that acutely precipitate exacerbation of congestive heart failure (CHF) in patients with left ventricular dysfunction. Knowledge of such factors is important in designing measures to prevent deterioration of clinical status. The objective of this study was to prospectively describe the precipitants associated with exacerbation of CHF status in patients enrolled in the Randomized Evaluation of Strategies for Left Ventricular Dysfunction Pilot Study. METHODS: We conducted a 2-stage, multicenter, randomized trial in 768 patients with CHF who had an ejection fraction of less than 40%. Patients were randomly assigned to receive enalapril maleate, candesartan cilexetil, or both for 17 weeks, followed by randomization to receive metoprolol succinate or placebo for 26 weeks. Investigators systematically documented information on clinical presentation, management, and factors associated with the exacerbation for any episode of acute CHF during follow-up. RESULTS: A total of 323 episodes of worsening of CHF occurred in 180 patients during 43 weeks of follow-up; 143 patients required hospitalization, and 5 died. Factors implicated in worsening of CHF status included noncompliance with salt restriction (22%); other noncardiac causes (20%), notably pulmonary infectious processes; study medications (15%); use of antiarrhythmic agents in the past 48 hours (15%); arrhythmias (13%); calcium channel blockers (13%); and inappropriate reductions in CHF therapy (10%). CONCLUSIONS: A variety of factors, many of which are avoidable, are associated with exacerbation of CHF. Attention to these factors and patient education are important in the prevention of CHF deterioration.

Adrenergic beta-Antagonists↗

Preparation of a chitin-apatite composite by in situ precipitation onto porous chitin scaffolds.

Composites of chitin with calcium phosphate were obtained by in situ precipitation of the mineral from a supersaturated solution onto chitin scaffolds. The chitin scaffolds were obtained by freeze drying to give a highly porous structure possessing a polar surface favorable for apatite nucleation and growth. THe extent and arrangement of calcium phosphate deposits on the chitin and substituted chitin scaffolds were explored. Up to 55% by mass of calcium phosphate could be incorporated into chitin scaffolds. Deposits on the chitin surface were a continuous apatite carpet nature while deposits on carboxymethylated chitin surfaces displayed a spherical morphology. Carboxymethylation of chitin exerts an overall inhibitory effect towards calcium phosphate deposition, but it provides for site-specific nucleation of the mineral phase. In situ precipitation can be an important route in the future production of various polymer-calcium phosphate composites.

Apatites↗

Specificity in systemic lupus erythematosus of antibodies to double-stranded DNA measured with the polyethylene glycol precipitation assay.

Recently, a new radioimmunoassay--the polyethylene glycol (PEG) assay--was introduced to measure antibodies to double-stranded (ds) DNA. In this method, polyethylene glycol precipitation of formed 3H-DNA/antiDNA complexes is used instead of the ammonium sulfate precipitation used in the Farr assay. In contrast to the Farr assay, with which only high-avidity antibodies to dsDNA are detected, the PEG assay also reportedly measures anti-dsDNA of relatively low avidity. We studied whether this gain in antibody measurement results in loss of specificity for systemic lupus erythematosus. When the PEG assay was applied to a selected panel of 440 sera from patients with various well-defined autoimmune diseases and to a group of 197 normal human control sera, matched sex and age to the patients, the method was found to be fairly specific for systemic lupus erythematosus, although the sera from some patients with myasthenia gravis and some with autoimmune liver disease were also found positive. Screening of 352 additional serum specimens, sent to our laboratory for diagnostic reasons, revealed that, with the PEG assay, an extra population of relatively low-avidity antibodies to dsDNA--missed by the Farr assay--was detected. Upon clinical evaluation, we found that the patients in whom such antibodies were detected generally fulfilled a number of the preliminary criteria of the American Rheumatism Association for systemic lupus erythematosus, but that this diagnosis often was not made. We claim that the presence of low-avidity antiDNA characterizes a milder form of the disease in which patients often show only a single clinical feature of the disease. We conclude that results of the PEG assay add valuable diagnostic and clinical information to results obtained by the Farr assay.

Antibody Specificity↗

Analysis of an engineered sulfate reduction pathway and cadmium precipitation on the cell surface.

We previously have genetically engineered an aerobic sulfate reduction pathway in Escherichia coli for the generation of hydrogen sulfide and demonstrated the pathway's utility in the precipitation of cadmium. To engineer the pathway, the assimilatory sulfate reduction pathway was modified so that cysteine was overproduced. Excess cysteine was then converted by cysteine desulfhydrase to an abundance of hydrogen sulfide, which then reacted with aqueous cadmium to form cadmium sulfide. In this study, observations of various E. coli clones were combined with an analysis of kinetic and transport phenomena. This analysis revealed that cysteine production is the rate-limiting step in the engineered pathway and provided an explanation for the phenomenon of cell surface precipitation. An analytical model showed that cadmium sulfide must form at the cell surface because the rate of cadmium sulfide formation is extremely fast and the rate of sulfide transport is relatively slow.

Cadmium↗

Liposome immunoblotting assay using a substrate-forming precipitate inside immunoliposomes.

A new immunoblotting assay which uses antibody-coupled liposomes containing horseradish peroxidase is proposed. A substrate 4-chloro-1-naphthol permeated through the phospholipid membrane of the antibody-coupled liposomes and formed a colored product precipitating inside the liposomes. The precipitates accumulated in the liposomes and could be detected at the positions where the liposomes coupled with a target in blotted samples. Combination of liposomes with average diameter of 350 nm and a PVDF membrane with a pore size of 450 nm, 0.02 ng of IgM was detected, while the conventional immunoblotting using antibody-HRP conjugates detected 2 ng of IgM. The sensitivity increased about two orders of magnitude by the liposome immunoblotting assay. This liposome immunoblotting assay gives a simple detection method of proteins with a high sensitivity, as well as a high sensitivity Western blotting assay.

Blotting, Western↗

Effects of hydraulic retention time and sulfide toxicity on ethanol and acetate oxidation in sulfate-reducing metal-precipitating fluidized-bed reactor.

The effects of hydraulic retention time (HRT) and sulfide toxicity on ethanol and acetate utilization were studied in a sulfate-reducing fluidized-bed reactor (FBR) treating acidic metal-containing wastewater. The effects of HRT were determined with continuous flow FBR experiments. The percentage of ethanol oxidation was 99.9% even at a HRT of 6.5 h (loading of 2.6 g ethanol L(-1) d(-1)), while acetate accumulated in the FBR with HRTs below 12 h (loading of 1.4 g ethanol L(-1) d(-1)). Partial acetate utilization was accompanied by decreased concentrations of dissolved sulfide (DS) and alkalinity in the effluent, and eventually resulted in process failure when HRT was decreased to 6.1 h (loading of 2.7 g ethanol L(-1) d(-1)). Zinc and iron precipitation rates increased to over 600 mg L(-1) d(-1) and 300 mg L(-1) d(-1), respectively, with decreasing HRT. At HRT of 6.5 h, percent metal precipitation was over 99.9%, and effluent metal concentrations remained below 0.08 mg L(-1). Under these conditions, the alkalinity produced by substrate utilization increased the wastewater pH from 3 to 7.9-8.0. The percentage of electron flow from ethanol to sulfate reduction averaged 76 +/- 10% and was not affected by the HRT. The lowest HRT did not result in significant biomass washout from the FBR. The effect of sulfide toxicity on the sulfate-reducing culture was studied with batch kinetic experiments in the FBR. Noncompetitive inhibition model described well the sulfide inhibition of the sulfate-reducing culture. (DS) inhibition constants (K(i)) for ethanol and acetate oxidation were 248 mg S L(-1) and 356 mg S L(-1), respectively, and the corresponding K(i) values for H(2)S were 84 mg S L(-1) and 124 mg S L(-1). In conclusion, ethanol oxidation was more inhibited by sulfide toxicity than the acetate oxidation.

Acetates↗

Selective precipitation and recovery of xylanase using surfactant and organic solvent.

The selective precipitation of xylanase from an aqueous phase containing mixtures of xylanase and cellulase was studied using an ionic surfactant as precipitating ligand and a polar organic solvent as recovery solvent. Of four ionic surfactants tested, sodium di-(2-ethylhexyl) sulfosuccinate (AOT) showed a complete removal of xylanase at pH 4.5. The recovery of xylanase from the xylanase-AOT complex was a strong function of the type and the volume of the polar solvent and of the concentration of sodium acetate buffer in the final aqueous solution used to solubilize the recovered xylanase. With ethanol as a recovery solvent, a recovery of xylanase activity of 78 +/- 10% was obtained. The cellulase activity in the recovered xylanase was below the detection limit. The results demonstrate that an ionic surfactant can recover enzymes from aqueous solutions without loss in their activity.

Cellulase↗

Precipitation by polycation as capture step in purification of plasmid DNA from a clarified lysate.

The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Large-scale purification of plasmid DNA from bacterial cell culture normally includes one or several chromatographic steps. Prechromatographic steps include precipitation with solvents, salts, and polymers combined with enzymatic degradation of nucleic acids. No method alone has so far been able to selectively capture plasmid DNA directly from a clarified alkaline lysate. We present a method for selective precipitation of plasmid DNA from a clarified alkaline lysate using polycation poly(N, N'-dimethyldiallylammonium) chloride (PDMDAAC). The specific interaction between the polycation and the plasmid DNA resulted in the formation of a stoichiometric insoluble complex. Efficient removal of contaminants such as RNA, by far the major contaminant in a clarified lysate, and proteins as well as 20-fold plasmid concentration has been obtained with about 80% recovery. The method utilizes a inexpensive, commercially available polymer and thus provides a capture step suitable for large-scale production.

Absorption↗

Light-responsive bioconjugates as novel tools for specific capture of biologicals by photoaffinity precipitation.

Light-responsive bioconjugates are synthesized by a two-step protocol calling first for cotelomerization (chain-transfer polymerization) of N-isopropylacrylamide and N-acryloxysuccinimide. The desired bioligand (biotin) is used in modified form as chain-transfer agent in this step. As a consequence, 100% of the produced bioconjugates carry this group. In a second step, the cotelomers (bioconjugates) are rendered photoresponsive by linking a chromophore ((3-aminopropyloxy)azobenzene) group to the N-acryloxysuccinimide side chains. The resulting structures show a critical solution temperature in pure water of 16 degrees C when the azo groups in the side chains are predominately in the (stable) trans state. Irradiation with UV light (330 nm) switches the azo group into the more hydrophilic cis state, and the critical solution temperature rises to 18 degrees C. Irradiation with visible light (> 440 nm) switches the group back to the trans state. Adjusting the temperature to an intermediate level, the bioconjugates are used to demonstrate the concept of photo affinity precipitation, i.e., the specific capture and recovery by light-induced precipitation of a target molecule (avidin) from a serum-containing cell-culture supernatant. The avidin was obtained in highly purified form; no nonspecific copurification of protein impurities was observable.

Acrylamides↗

Direct detection of carrier ampholytes in immobilized pH gradients using picric acid precipitation.

A protocol is described for monitoring the heterogeneity of end products of organic syntheses yielding amphoteric molecules containing two or more amino groups. This protocol was found to be a valuable aid in synthesis of carrier ampholytes for specific isoelectric focusing applications. This method does not depend on the ampholytes themselves to dictate the conditions under which they are analyzed. Carrier ampholytes have been found previously to be insoluble in picric acid and the insolubility property was not dependent upon the pI of individual ampholyte species. This insolubility property was exploited in the protocol. Immobilized pH gradients were used to focus the carrier ampholytes. Ampholytes were then visualized in situ by picric acid precipitation. The data shows that the protocol is useful for analyzing the results of chemical manipulations for enhancing the resolution of carrier ampholytes. A direct relationship was shown between carrier ampholyte heterogeneity as demonstrated by this protocol and the resolution of complex protein mixtures in isoelectric focusing gels. Picric acid formed visible precipitates with a variety of organic compounds which contained more than one amino group.

Acrylamides↗

An antibody which precipitates Dane particles in acute hepatitis type B: relation to receptor sites which bind polymerized human serum albumin on virus particles.

An antibody, which is distinct from the HBsAg- reacts with antigenic sites on Dane particles- HBcAg and HBeAg, was studied by radioimmunoprecipitation of radioactive intact hepatitis B virions in sera obtained early in the course of acute hepatitis type B. The antibody, previously termed anti-Dane particle (anti-DP) antibody, was reactive with Dane particles and HBsAg particles obtained from HBeAg-positive sera but not with HBsAg particles from anti-HBe containing sera. The expression on virus particles of the evoking antigen correlated with levels of binding sites for polymerized human serum albumin (pHSA) as detected by solid-phase radioimmunoassay. In acute hepatitis B sera, levels of anti-DP antibody activity showed inverse correlation with expression of pHSA receptors on circulating virus particles, although the two reactivities were not mutually exclusive. In inhibition experiments, pHSA blocked precipitation of Dane particles by anti-DP positive sera, while native human albumin and polymerized bovine albumin had no effect. The inhibition by pHSA of the anti-DP reaction appeared specific since identical concentrations of pHSA did not interfere with precipitation of virus particles by anti-HBs. Affinity chromatography studies with anti-DP insolubilized on Sepharose 4B columns showed selective binding to the gel of radioactive Dane particles; 125I-HBsAg was not reactive. The binding of Dane particles to anti-DP columns was completely inhibited when virus particles were applied to the gel in pHSA; pretreatment of the column with pHSA did not affect the reaction.(ABSTRACT TRUNCATED AT 250 WORDS)

Centrifugation, Density Gradient↗

Evaluation of a precipitation hardened wrought cobalt-nickel-chromium-titanium alloy for surgical implants.

A Co-Ni-Cr-Ti precipitation hardening alloy is evaluated for use as a surgical implant alloy. Static and fatigue strength studies indicate that the alloy is equivalent, or superior to, present implant alloys. In vitro and in vivo corrosion studies indicate that the material has good general corrosion resistance but may be susceptible to crevice corrosion. However, there appear to be no inherent difficulties in using precipitation hardened alloys for surgical implants and therefore it may be possible to develop a new class of materials possessing ultra-high strength and excellent corrosion resistance for use in surgical implant applications.

Alloys↗

In vitro blood compatibility of glycosaminoglycan-precipitated collagens.

Precipitation of bovine hide collagen by chondroitin 6-sulfate at low pH and subsequent crosslinking enhances the blood compatibility of native collagen. Both dehydrothermal crosslinking and complexation with chrondroitin 6-sulfate separately decrease the platelet-aggregating activity of collagen. Crosslinking also decreases the number of free acidic and free basic residues on collagen, which suggests that crosslinking involves these residues in condensation reactions with formation of intrachain and interchain synthetic peptide bonds. Clotting times for collagen precipitated with chondroitin 6-sulfate indicate that this surface does not activate or interfere with coagulation via either the intrinsic or extrinsic pathway. These findings support further consideration of collagen modified by chondroitin 6-sulfate as a blood compatible material.

Animals↗

Fabrication and characterization of hydrophilized porous PLGA nerve guide conduits by a modified immersion precipitation method.

Nerve guide conduits (NGCs) with selective permeability and hydrophilicity were fabricated using poly(lactic-co-glycolic acid) (PLGA) and Pluronic F127 by a modified immersion precipitation method developed by our laboratory. The hydrophilized porous PLGA tubes as NGCs were fabricated by immersing a water-saturated rod-shape alginate hydrogel into PLGA/Pluronic F127 mixture solution (in tetraglycol). The PLGA/Pluronic F127 mixture was precipitated outside the alginate hydrogel rod by the diffusion of water from the hydrogel rod into PLGA/Pluronic F127 mixture solution. The inner diameter and wall thickness of tubes could be easily controlled by adjusting the diameter of alginate hydrogel rod and immersion time, respectively. It was observed that the tube wall has an asymmetric column-shape porous structure. The inner surface of the tube had nano-size pores ( approximately 50 nm), which can effectively prevent from fibrous tissue infiltration but permeate nutrients and retain neurotrophic factors, while the outer surface had micro-size pores ( approximately 50 microm), which can allow vascular ingrowth for effective supply of nutrients and oxygen into the tube. From the investigations of mechanical property, water absorbabiliy, and model nutrient permeability of the tubes, the hydrophilized PLGA/F127 (3 wt %) tube seems to be a good candidate as a NGC for the effective permeation of nutrients as well as the good mechanical strength to maintain a stable support structure for the nerve regeneration.

Chemical Precipitation↗

Investigation of ion-pair precipitates of selected alkoxylates and complex salts of specific metal cations by liquid secondary ion mass spectrometry.

Liquid secondary ion (LSI) mass spectra of ion-pair precipitates obtained for Triton X-100 with strontium, lead, cadmium and mercury tetraphenylborates and for selected butoxylene-ethoxylene monoalkyl ethers with barium tetraiodobismuthate(III) are discussed. On the basis of LSI mass spectra, recorded in both positive and negative modes, the formulae of the ion-pair precipitates were determined. On the basis of B/E mass spectra, the fragmentation routes of [M - H + Ba](+) ions for butoxylene-ethoxylene monoalkyl ether complexes of barium and [M - H + Cd](+) ions for the Triton X-100 complex of cadmium are proposed.

Barium↗

Prediction of precipitation-induced phlebitis: a statistical validation of an in vitro model.

To avoid phlebitis, new intravenous (IV) parenterals are often screened by injection into animals. This method is not only expensive and time consuming, it is also detrimental to the animals. An alternate method, focusing on precipitation as the cause, uses an in vitro dynamic injection model that requires less money and time and reduces the need for live models. Validation of the dynamic injection apparatus, for predicting mechanical phlebitis, is established. Twenty-one currently marketed IV products were injected into isotonic Sorenson's phosphate buffer flowing at 5 mL/min. The resulting opacities, produced by precipitation, are measured in an ultraviolet flow cell. These opacity data, coupled with literature reports on phlebitis occurrence, were used to generate a logistic regression that indicates the probability of phlebitis given an opacity value measured by the apparatus. Regression results are supported by a receiver operator characteristic curve that establishes the most ideal cut-off opacity value. This opacity value provides the highest combined sensitivity (statistical power) and specificity while minimizing false-positive and false-negative results. Both analyses show that an opacity value of 0.003 best delineates phlebitic and nonphlebitic products. Measures of sensitivity (0.83), specificity (0.93), positive predictive value (0.93), and negative predictive value (0.78) indicate the model's predictive accuracy and reliability. These results support the use of the dynamic model in place of animals for preliminary phlebitis testing of new IV injectables.

Chemical Precipitation↗