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Organic nitrogen in precipitation: real problem or sampling artefact?

Published observations of organic nitrogen (N) compounds in precipitation go back almost a century. Several different methods have been used to measure both the total and ionic concentrations of N. There is therefore some uncertainty as to whether reported "organic N" is real, or simply the result of uncertainties in chemical analyses or inadequate sampling methods. We found that the materials from which the collector was made (polypropylene, steel, or glass) had no significant effect on the composition of dissolved organic N (DON). The use of a biocide was found to be very important during sampling and storage of samples before analysis. We set up a network of seven collectors across the U.K., from the Cairngorms to Dorset, all operating to the same protocol, and including a biocide. Samples were analysed centrally, using proven methods. Over 6 months, organic N contributed about 20% to the total N in U.K. precipitation, but with a large variation across the country. This means that current estimates of wet deposited N to the U.K., which are based only on the ammonium and nitrate concentrations, are too small. Organic N is not an artefact, but a real problem that needs to be addressed.

Air Pollutants↗

Further characterization of the ovine lutropin alpha and beta subunits prepared by the salt precipitation method.

The subunits of ovine lutropin prepared by acid dissociation and salt precipitation were characterized by end group analysis, tryptic peptide mapping, SDS gel electrophoresis and biological activity. No evidence of internal peptide cleavage was found in the alpha subunit. The subunits possessed low activity. The alpha and beta subunits recombined effectively to generate a complex that had full receptor binding activity and in vitro biological activity. The recombinants of subunits prepared by countercurrent distribution showed only 50% activity in both assays. The salt precipitation method alpha subunit could be completely reduced and reoxidized in the absence of denaturants. The reoxidized alpha subunit combines with the native beta subunit generating full activity. However, this recombined hormone tends to lose activity with time, suggesting that the reoxidation may not fully restore the native structur of the reduced alpha subunit. The native lutropin alpha subunit effectively combined with follitropin beta subunit generating complete follitropin activity.

Amino Acid Sequence↗

Complete primary structure of statherin, a potent inhibitor of calcium phosphate precipitation, from the saliva of the monkey, Macaca arctoides.

Human saliva, which is supersaturated with respect to basic calcium phosphate salts, is stabilized primarily by the presence of two classes of phosphoproteins, statherin and the acidic proline-rich proteins (PRP). These molecules act by inhibiting both primary (spontaneous) precipitation of calcium phosphates in saliva and secondary (surface induced) precipitation of these salts onto dental enamel. The complete amino-acid sequences of several human PRP and the N-terminal sequence of PRP from saliva of M. arctoides have been determined. Similarly, the complete sequence of statherin from human and M. fascicularis saliva is known. We now report the complete structure of statherin from the saliva of the stump-tailed monkey, M. arctoides. The structure was determined by gas-phase sequencing of intact statherin, elucidating positions 1-26, and sequencing an unpurified mixture of tryptic peptides which elucidated the remaining positions through the C-terminus (residue 42) of the molecule. This latter degradation produced an eight amino-acid overlap with that of intact statherin and was confirmed by C-terminal analysis and amino-acid composition of native statherin. The complete amino-acid sequence of M. arctoides statherin is: NH2-Asp-PSer-PSer-Glu-Glu5-Lys-Phe-Leu-Arg-Arg10 -Leu-Arg-Arg-Phe-Asp15-Glu- Gly-Arg-Tyr-Gly20-Pro-Tyr-Gln-Pro-Phe25-Val-Pro-Pro- Pro29Leu30-Tyr- Pro-Gln-Pro-Tyr35-Gln-Pro-Tyr-Gln-Pro40-Gln-Tyr-COOH This sequence differs from human statherin at positions 11, 12, 15, 16, 18, 25-27, 38-40 and from M. fascicularis statherin at positions 26 and 28.

Amino Acid Sequence↗

Separation of two hepatitis B human antibody fractions by cold-ethanol precipitation.

Antibodies to three different samples of hepatitis B antigen-positive serum were found to be absent from a Cohn fraction II hepatitis B immune globulin preparation, but present in the whole plasma from which the immune globulin was made. Fractions containing the antibodies specific for the three antigen samples were isolated from the plasma by euglobulin precipitation, and the antibodies were characterized as to immunoglobulin class and precipitability in the Cohn fractionation procedure. They appear to be IgG antibodies which, however, were detectable only in Cohn fraction I, while the remainder of the HB specificity present in the original plasma was found, as expected, in fraction II.

Antibodies↗

Characterization of bovine anterior-lens-capsule basement-membrane collagen. 2. Segment-long-spacing precipitates: further evidence for large N-terminal and C-terminal extensions.

Collagen extracted from bovine anterior lens capsule basement membrane under minimally degradative conditions can be precipitated in the form of segment-long-spacing crystallites. Positive and negative stain preparations of these precipitates, using uranyl acetate, show the crystallites to have an asymmetric band pattern, indicating unidirectional orientation of all molecules in each crystallite. Non-banded globular masses appear at both ends of most of the crystallites, giving the segment-long-spacing aggregates the form of dumbells, with a total length of 300 nm. Repeated pepsin digestion removes first one then both globular regions leaving the helical region intact but with a length still equal to 300 nm. It thus appears that the pepsin-susceptibile protein is folded back over and covers the ends of the helical regions. Preparations with the most intact end regions tend to aggregate in fibrous segment-long-spacing form, that is, in end-interacted strings or nets. The very prominent molecular end-region interaction suggests a similar filamentous but non-periodic organization within native basement membrane.

Animals↗

Heterogeneity of brush-border-membrane vesicles from rat small intestine prepared by a precipitation method using Mg/EGTA.

Brush border membrane vesicles from rat small intestine were isolated by a Mg/EGTA precipitation method. Further fractionation either by free flow electrophoresis or by sucrose density gradient centrifugation leads to subfractions which differ with respect to enzyme enrichment factors, transport properties for D-glucose and protein pattern analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. A relative enrichment of (Na+ + K+)-ATPase is found in one fraction, whereas in another fraction maltase, aminopeptidase M and alkaline phosphatase are relatively enriched. The fractions show different properties of D-glucose transport under tracer exchange conditions and a different inhibition of D-glucose transport by phlorizin and phloretin. These results indicate that the vesicles obtained from rat small intestine by this cation precipitation method are not homogeneous. The inhomogeneity cannot be due to a crosscontamination by membranes other than from the cell envelopment, as none of the fractions show a significant enrichment of succinate--cytochrome c oxidoreductase, KCN-resistant NADH oxidoreductase or glucosaminidase. The inhomogeneity might be due either to a crosscontamination by basal-lateral membranes or to membranes derived from epithelial cells not yet fully differentiated.

Animals↗

Isolation of a novel antibody, which precipitates ribonucleoprotein complex containing threonine tRNA from a patient with polymyositis.

We have isolated and characterized a novel antibody from a patient with polymyositis, which precipitates threonine tRNA and an unknown small RNA molecule from 32P-labeled HeLa extract. Fingerprint analysis showed that the human threonine tRNA precipitated was nearly identical to the murine tRNA described by Harada [(1978) Seikagaku 50, 397-411]. Nucleotide analysis suggested that the other small RNA molecule might be transfer RNA. Since phenol extraction resulted in the loss of immunoprecipitability, the protein portion was presumed to be involved in the recognition of the antigen. Immunofluorescence staining of HeLa cells with the antibody clearly demonstrated that the antigen was located predominantly in the cytoplasm.

Autoantibodies↗

Proteomic analysis of normal human urinary proteins isolated by acetone precipitation or ultracentrifugation.

BACKGROUND: Proteomic techniques have recently become available for large-scale protein analysis. The utility of these techniques in identification of urinary proteins is poorly defined. We constructed a proteome map of normal human urine as a reference protein database by using two differential fractionated techniques to isolate the proteins. METHODS: Proteins were isolated from urine obtained from normal human volunteers by acetone precipitation or ultracentrifugation, separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and identified by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry followed by peptide mass fingerprinting. RESULTS: A total of 67 protein forms of 47 unique proteins were identified, including transporters, adhesion molecules, complement, chaperones, receptors, enzymes, serpins, cell signaling proteins and matrix proteins. Acetone precipitated more acidic and hydrophilic proteins, whereas ultracentrifugation fractionated more basic, hydrophobic, and membrane proteins. Bioinformatic analysis predicted glycosylation to be the most common explanation for multiple forms of the same protein. CONCLUSIONS: Combining two differential isolation techniques magnified protein identification from human urine. Proteomic analysis of urinary proteins is a promising tool to study renal physiology and pathophysiology and to determine biomarkers of renal disease.

Acetone↗

Aluminium allergy in patients hyposensitized with aluminium-precipitated antigen extracts.

During hyposensitization therapy with aluminium-precipitated antigen solutions, a small % of patients develop persistent subcutaneous nodules at the injection site; the existence of delayed sensitivity to aluminium has been implicated in the pathogenesis of these nodules. We studied the prevalence of aluminium sensitivity (using patch, prick and intradermal tests) and common contact allergens (TRUE Test) in 20 healthy subjects, and in 40 patients treated with aluminium-containing extracts, 20 of whom had persistent subcutaneous nodules that remained for more than 2 months, the other half having no nodular reactions or nodules that remained for less than 2 months. Aluminium sensitivity was found only in those patients of the treated group who had persistent nodular reactions, 4 cases of positivity to an aluminium chloride patch test being found. All 4 cases were women, nodules remained for more than 6 months, and intracutaneous tests were negative. 3 of them also had contact sensitivity to nickel. In 2 cases, nodules were removed for histological and histochemical examination, showing non-specific inflammatory granulomas, and aluminium crystals being found in only 1 case. It is concluded that delayed sensitivity to aluminium appears to be implicated in the pathogenesis of persistent nodular reactions, but sensitivity to aluminium was not found in patients treated with aluminium-precipitated extracts without persistent nodular reactions.

Adolescent↗

The role of blood platelets in the precipitation of soluble fibrin by endotoxin.

This study is intended to clarify whether or not platelets are essential to endotoxin-induced precipitation of soluble fibrin representing the second phase of microclot formation. Renal glomerular microclots could be observed in extremely thrombocytopenic rabbits to the same extent as in normal rabbits after the injection of endotoxin into animals with circulating soluble fibrin. Thrombocytopenia was induced by feeding rabbits busulphan, and soluble fibrin was produced by infusing ancrod. The experiments indicate that platelets are not essential to the endotoxin-induced precipitation of soluble fibrin.

Animals↗

Die-spacing technique by diffusion precipitation.

The present study has shown that it is possible by utilizing the principle of diffusion to precipitate on stone dies an adequate resin spacer of uniform and desirable thickness. Details and variables of the technique are described. The precipitation technique is simple and requires a minimum of time and equipment. This spacer technique opens up gateways for a new and improved casting technique cleared of most of the variables affecting the precision of the methods employed up till now. A new gypsum bonded investment for the new casting technique is being developed.

Acetone↗

Inhibition of precipitation in counter current electrophoresis. A sensitive method for detection of mink antibodies to Aleutian disease virus.

Inhibition of precipitation in counter current electrophoresis was at least 32 more sensitive when compared to the normal counter current electrophoresis for the demonstration of mink antibodies against Aleutian disease virus (ADV). ADV antigen can be produced from mink organs or in cell culture. The reactivity of the two types of antigen in the two kinds of counter current electrophoresis methods is described in this report. When 22 mink sera were titrated in normal counter current electrophoresis against cell culture produced antigen and organ produced antigen, significantly lower antibody titres were found with cell culture produced antigen. The difference was not found when inhibition of precipitation of counter current electrophoresis was used.

Aleutian Mink Disease↗

Some factors affecting the release of imipramine from gel-precipitated aluminium hydroxide spheres.

Changing the pH of the dissolution medium has been found to affect the release of imipramine from gel-precipitated aluminium hydroxide spheres. Release from unwashed, unheated spheres into solutions of pH 1.2 was controlled by dissolution of the gel matrix, whereas that into solutions of pH 3 and pH 5 appeared to be under diffusion control. The liberation of drug from unwashed, heated spheres into the media of higher pH exhibited more complex kinetics. Washed spheres failed to release significant amounts of imipramine into the solutions of pH 3 and 5. Changing the ionic strength of the media had little effect on drug release. These phenomena have been explained with reference to model theories of the precipitation and ageing of aluminum hydroxide gels and their pH-solubility profiles.

Aluminum Hydroxide↗

Vinblastine-induced precipitation of microtubule protein.

Vinblastine quantitatively precipitates a protein from supernatants obtained from high-speed centrifugation of homogenates of HeLa cells and of pig brain. This protein migrates as a single band on gel electrophoresis, has a mobility identical to that of purified microtubule protein, and-like microtubule protein-binds colchicine. The precipitation is partially inhibited by 0.9 percent NaCl.

Animals↗

Plasmid-encoded copper resistance and precipitation by Mycobacterium scrofulaceum.

A copper-tolerant Mycobacterium scrofulaceum strain was able to remove copper from culture medium by sulfate-dependent precipitation as copper sulfide. Such precipitation of copper sulfide was not observed in a derivative that lacks a 173-kilobase plasmid. In addition, the plasmid-carrying strain has a sulfate-independent copper resistance mechanism.

Chemical Precipitation↗

A new Klebsiella planticola strain (Cd-1) grows anaerobically at high cadmium concentrations and precipitates cadmium sulfide.

Heavy metal resistance by bacteria is a topic of much importance to the bioremediation of contaminated soils and sediments. We report here the isolation of a highly cadmium-resistant Klebsiella planticola strain, Cd-1, from reducing salt marsh sediments. The strain grows in up to 15 mM CdCl(2) under a wide range of NaCl concentrations and at acidic or neutral pH. In growth medium amended with thiosulfate, it precipitated significant amounts of cadmium sulfide (CdS), as confirmed by x-absorption spectroscopy. In comparison with various other strains tested, Cd-1 is superior for precipitating CdS in cultures containing thiosulfate. Thus, our results suggest that Cd-1 is a good candidate for the accelerated bioremediation of systems contaminated by high levels of cadmium.

Anaerobiosis↗

Reductive precipitation of gold by dissimilatory Fe(III)-reducing bacteria and archaea.

Studies with a diversity of hyperthermophilic and mesophilic dissimilatory Fe(III)-reducing Bacteria and Archaea demonstrated that some of these organisms are capable of precipitating gold by reducing Au(III) to Au(0) with hydrogen as the electron donor. These studies suggest that models for the formation of gold deposits in both hydrothermal and cooler environments should consider the possibility that dissimilatory metal-reducing microorganisms can reductively precipitate gold from solution.

Archaea↗

Simple and reliable method to precipitate proteins from bacterial culture supernatant.

A simple and reliable method for precipitating protein from bacterial culture supernatants based on a pyrogallol red-molybdate-methanol (PRMM) protocol has been developed and applied for the analysis of proteins secreted by a bacterial type III secretion system. PRMM-based precipitation has been shown to be more efficient and robust than are conventional protocols.

Bacterial Proteins↗