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PubMed2005

AgCl precipitates in isolated cuticular membranes reduce rates of cuticular transpiration.

Counter diffusion of chloride, applied as NaCl at the inner side of isolated cuticles, and silver, applied as AgNO(3) at the outer side, lead to the formation of insoluble AgCl precipitates in isolated cuticles. AgCl precipitates could be visualized by light and scanning electron microscopy. The presence of AgCl precipitates in isolated cuticles was verified by energy dispersive X-ray analysis. It is argued that insoluble AgCl precipitates formed in polar pores of cuticles and as a consequence, cuticular transpiration of 13 out of 15 investigated species was significantly reduced up to three-fold. Water as a small and uncharged but polar molecule penetrates cuticles via two parallel paths: a lipophilic path, formed by lipophilic cutin and wax domains, and a aqueous pathe, formed by polar pores. Thus, permeances P (m s(-1)) of water, which is composed of the two quantities P (Lipid) and P (Pore), decreased, since water transport across polar pores was affected by AgCl precipitates. Cuticles with initially high rates of cuticular transpiration were generally more sensitive towards AgCl precipitates compared to cuticles with initially low rates of transpiration. Results presented here, significantly improves the current model of the structure of the cuticular transpiration barrier, since the pronounced heterogeneity of the cuticular transport barrier, composed of lipophilic as well as polar paths of diffusion, has to be taken into account in future.

Chemical Precipitation↗

PubMed1984

Precipitation of collagens by polyethylene glycols.

Types I, II, and III collagens are readily precipitated at neutral pH by polyethylene glycols (PEG). As the molecular weight fraction of the polyethylene glycols increases, they become more effective as precipitants on a weight basis. The amount of PEG required for precipitation depends on the pH, the ionic strength, and the nature of the buffer or salts present. In tissue culture media, low concentrations of collagens and procollagens may be quantitatively precipitated and readily collected by low-speed centrifugation. Polyethylene glycol precipitation can be used to obtain collagens and procollagens from tissue culture media at either analytical or preparative scale, and since the polyethylene glycols do not bind to collagens, the precipitates may be further analyzed directly by chromatographic or electrophoretic methods.

Cells, Cultured↗

PubMed1988

Precipitation of calcium palmitate from bile salt-containing dispersions.

Addition of calcium chloride to mixed micellar systems composed of sodium salts of palmitic acid and high concentrations of different bile acids results in precipitation of Ca(palmitate)2 only when the palmitate concentration exceeds a critical value, which is dependent on the concentrations of Ca2+, Na+ and bile salt, and on the type of bile salt used. All these dependencies, as well as the complex and interrelated effects of the various parameters on the kinetics of Ca(palmitate)2 precipitation are consistent with the following mechanism: (i) calcium binds to palmitate-bile salt mixed micelles and promotes their aggregation, at a rate governed by the concentration ratio between bound calcium and micelles (here denoted "binding ratio"). (ii) Ca(palmitate)2 precipitation occurs within the aggregate of micelles only if those micelles include sufficient amounts of Ca2+ and palmitate to allow for the formation of large enough crystal units of Ca(palmitate)2 which can serve as nucleation "seeds". Both the concentrations of micelles and Na+ have dual effects on the rate of precipitation. Increasing micelle concentration, by itself, accelerates aggregation but at the same time leads to a decrease of the binding ratio, thus reducing the rate of precipitation. Na+ which reduces the binding ratio through competitive binding also reduces the surface charge, thus assisting micelle aggregation. Our model also explains the facilitation of precipitation observed when phosphatidylcholine is contained in the palmitate-bile salt mixed micelles and the inhibitory effect of the water soluble bovine serum albumin.

Bile Acids and Salts↗

PubMed1979

Optimization of parameters in protein nitrogen unit precipitation procedure for allergenic extracts.

This work establishes the optimum conditions for the protein nitrogen unit (PNU) phosphotungstic acid (PTA) precipitation procedure for allergenic extracts. The volume of extract analyzed, the dilution of the extract prior to precipitation, the percent PTA in the precipitating solution, the washing of the precipitate, and the volume of concentrated HCl initially added to the extract were optimized, as they were found to be the factors which affected the amount of PTA precipitate obtained. Varying the digestion time, the digestion temperature, and the percent HCl in the 15% PTA precipitating solution did not cause detectable differences in the PNU values of the extracts studied.

Allergens↗

PubMed1989

Salt precipitation during the freeze-concentration of phosphate buffer solutions.

Salt precipitation during the freeze concentration of phosphate solutions was investigated by differential scanning calorimetry (DSC), in view of its practical importance in the cryopreservation or freeze-drying of biological materials. It was found that the fraction of salt precipitated depends on the initial salt concentration; it began to decrease with decreasing concentration at approx. 1 M. Salt precipitation also depends on the cooling rate. In some cases, cooling at approx. 10(3) degree min-1 inhibited salt precipitation which had been observed during slow cooling (0.62 degree min-1), without, however, affecting the shape of the ice melting endotherm. In the case of ternary phosphate buffers, the fraction of salt precipitating depends on the salt composition as well as the initial concentration and cooling rate. Near the composition of the ternary eutectic or the composition where two salts are present at the same concentration, salts were prevented from precipitation.

Buffers↗

PubMed1994

Differences of inner and outer hair cells in the organ of Corti of the guinea pig in respect to the cellular content of precipitable calcium.

Differences between inner and outer hair cells in the cellular content of precipitable calcium were detected using a potassium pyroantimonate precipitation method and the electron spectroscopic imaging (ESI-) technique. The cytoplasm of the inner hair cells was scattered with a high number of calcium precipitates in all analysed animals, but only a few reaction products could be identified in the cytoplasm of the outer hair cells in all analyzed specimens. Even the well developed system of the subsurface fenestrated cisternae in the outer hair cells was nearly empty of calcium precipitates. A relatively high amount of reaction products could be identified in the nuclei of both types of nerve endings of the receptor cells. Significant differences regarding the content of precipitable calcium were found in the different types of nerve endings, which come into contact with the basal parts of both receptor cells. The observed differences in the content of precipitable calcium between the two types of hair cells are discussed with respect to their probable different roles in signal transduction processes.

Animals↗

PubMed2000

Precipitation of specific proteins by freeze-thawing of human saliva.

Frozen saliva samples demonstrate a variable amount of precipitate on thawing depending on the type of secretion [submandibular-sublingual (SML) greater than parotid]. This precipitate has been resuspended using EDTA or removed by centrifugation by some workers and others do not mention it. Yet others collect the salivas into EDTA or centrifuge them before freezing. To determine the adsorption of proteins to hydroxyapatite, prior treatment with EDTA would be disadvantageous. The aim here was to determine if the protein pattern in parotid and SML saliva as demonstrated by sodium dodecyl sulphate gel electrophoresis is affected by the formation of precipitates. Portions of parotid and SML saliva were thawed and treated in the following ways: (a) mixed vigorously with a vortex mixer; (b) centrifuged to remove the precipitate; (c) mixed with EDTA (1 and 5 mmol final concentration for parotid and SML samples, respectively) to resuspend the precipitate. The samples were loaded on to gradient (5-20%) SDS gels and, following electrophoresis, the gels were stained with Coomassie brilliant blue R-250. The protein patterns obtained for (a) and (c) were the same. The centrifuged samples demonstrated loss of a specific band of less than 14 kDa, although this was less obvious in the parotid samples. The SML samples also showed a reduction in other lower molecular-weight proteins. This study demonstrates that precipitates in thawed frozen salivas contain specific proteins and that these samples require careful handling to avoid any alteration in the overall protein composition.

Chemical Precipitation↗

PubMed1998

The precipitation of mucin by aluminium.

The interactions of Al with a mucin glycopeptide have been studied. A number of specific reactions were identified the nature of which were dependent upon the Al chemistry in the hydration environment. In particular, Al was observed to precipitate mucin and it is suggested that this proceeded via the intercalation of the hydroxide within the hydrated macroreticular network of the mucin biopolymer. This precipitation of mucin was visible by eye and abolished the viscosity of native mucin. Viscometry indicated that Al was bound by mucin at low pH. At pH > 3 Al formed a low molecular weight complex with mucin which was hydrolytically stable and was not precipitated at pH up to 8. In an additional and competitive reaction Al was bound by mucin and the resultant mucin-Al complex was suggested to be the precursor to self-assembled mucin-Al spheres identified in solution, by photon correlation spectroscopy, and in precipitate using selective histochemistry. The majority of these spherical structures were of sub-micron diameter and, through their interaction with each other, were probably responsible for the observed pH-dependent peaks of mucin solution viscosity. The larger spheres, between 20 and 80 microns in diameter, were only identified in isolated mucin/Al precipitates and, being comparatively rare, were unlikely to have influenced solution viscosities. These large spheres were observed to act as possible nucleation sites for the flocculation of mucin/Al precipitate. Al at concentrations as low as 0.015 mM induced changes in the rheological properties of mucin. Considering the ubiquitous nature of mucin and the degree to which it is conserved within biota the interactions of Al with mucin may have wide ranging implications for biological systems.

Aluminum↗

PubMedDate not supplied

Experimental and numerical simulation of dissolution and precipitation: implications for fracture sealing at Yucca Mountain, Nevada.

Plugging of flow paths caused by mineral precipitation in fractures above the potential repository at Yucca Mountain, Nevada could reduce the probability of water seeping into the repository. As part of an ongoing effort to evaluate thermal-hydrological-chemical (THC) effects on flow in fractured media, we performed a laboratory experiment and numerical simulations to investigate mineral dissolution and precipitation under anticipated temperature and pressure conditions in the repository. To replicate mineral dissolution by vapor condensate in fractured tuff, water was flowed through crushed Yucca Mountain tuff at 94 degrees C. The resulting steady-state fluid composition had a total dissolved solids content of about 140 mg/l; silica was the dominant dissolved constituent. A portion of the steady-state mineralized water was flowed into a vertically oriented planar fracture in a block of welded Topopah Spring Tuff that was maintained at 80 degrees C at the top and 130 degrees C at the bottom. The fracture began to seal with amorphous silica within 5 days.A 1-D plug-flow numerical model was used to simulate mineral dissolution, and a similar model was developed to simulate the flow of mineralized water through a planar fracture, where boiling conditions led to mineral precipitation. Predicted concentrations of the major dissolved constituents for the tuff dissolution were within a factor of 2 of the measured average steady-state compositions. The mineral precipitation simulations predicted the precipitation of amorphous silica at the base of the boiling front, leading to a greater than 50-fold decrease in fracture permeability in 5 days, consistent with the laboratory experiment.These results help validate the use of a numerical model to simulate THC processes at Yucca Mountain. The experiment and simulations indicated that boiling and concomitant precipitation of amorphous silica could cause significant reductions in fracture porosity and permeability on a local scale. However, differences in fluid flow rates and thermal gradients between the experimental setup and anticipated conditions at Yucca Mountain need to be factored into scaling the results of the dissolution/precipitation experiments and associated simulations to THC models for the potential Yucca Mountain repository.

Chemical Precipitation↗

PubMed1999

Buffer capacity and precipitation control of pH solubilized phenytoin formulations.

The main objective of this investigation is to develop a phenytoin (DPH) intravenous formulation that does not precipitate upon dilution. The effect of the buffer capacity at pH 12 of several DPH formulations on the extent and lag-time of DPH precipitation upon dilution with Sorensen's phosphate buffer (SPB) is evaluated. DPH precipitation was evaluated by means of static and dynamic in vitro dilution methods. It is shown that an increase in the formulation buffer capacity decreases substantially the extent of DPH precipitation and increases the lag-time for precipitation. In addition, a comparison between static and dynamic in vitro methods to measure precipitation is presented.

Anticonvulsants↗

PubMed1998

Spruce budworm elastase precipitates Bacillus thuringiensis delta-endotoxin by specifically recognizing the C-terminal region.

A gut juice protein from Choristoneura fumiferana (spruce budworm) larvae that precipitates certain delta-endotoxins shows a unique specificity for the C-terminal amino acid sequence. Using homolog scanning mutants, we have identified a contiguous region of the Cry1Aa toxin which interacts with the 75-kDa toxin precipitating protein (TPP-75)' resulting in precipitation. The contiguous region from Cry1Aa can be transferred to Cry1Ac and results in an identical precipitation reaction. The precipitation reaction occurs rapidly and is unique in that the ratio of precipitating protein to toxin is low (estimated at 0.01), unlike antibody-antigen reactions which exhibit mole ratios close to 1. TPP-75 has been characterized as an elastase-like serine protease. We have taken advantage of this serine protease character and incorporated a radiolabel using an irreversible inhibitor. The radiolabel has allowed us to show the coincidence of the catalytically-inhibited TPP-75 with the toxin in a blotting assay and to follow the degradation of TPP-75 during storage. TPP-75 represents the first evidence that gut juice proteins may selectively attenuate the activity of delta-endotoxins, prior to binding to putative receptors on susceptible cells. TPP-75 should be evaluated as a possible resistance mechanism for those larvae that do not exhibit a receptor-based resistance.

Animals↗

PubMed2003

Enhancing protein crystallization through precipitant synergy.

Suitable conditions for protein crystallization are commonly identified by screening combinations of independent factors that affect crystal formation. Because precipitating agents are prime determinants of crystallization, we investigated whether a systematic exploration of combinations of mechanistically distinct precipitants would enhance crystallization. A crystallization screen containing 64 precipitant mixtures was devised. Tests with ten HIV envelope-related proteins demonstrated that use of precipitant mixtures significantly enhanced both the probability of crystallization as well as the quality of optimized crystals. Tests with hen egg white lysozyme generated a novel C2 crystal from a salt/organic solvent mixture; structure solution at 2 A resolution revealed a lattice held together by both hydrophobic and electrostatic dyad interactions. The results indicate that mechanistically distinct precipitants can synergize, with precipitant combinations adding unique dimensions to protein crystallization.

Animals↗

PubMed2004

Nanoscale precipitation in a maraging steel studied by APFIM.

This article summarizes findings from our previous investigations and recent studies concerning precipitation in a maraging steel of type 13Cr-9Ni-2Mo-2Cu (at.%) with small additions of Ti (1 at.%) and Al (0.7 at.%). The material was investigated after aging at 475 degrees C up to 400 h using both conventional and three-dimensional atom-probe analyses. The process of phase decomposition in the steel proved to be complicated. It consisted of precipitation of several phases with different chemistry. A Cu-rich phase was first to precipitate and Mo was last in the precipitation sequence. The influence of the complex precipitation path on the material properties is discussed. The investigation clearly demonstrated the usefulness of the applied techniques for investigation of nanoscale precipitation. It is also shown that, complementary methods (such as TEM and EFTEM) giving structural and chemical information on a larger scale must be applied to explain the good properties of the steel after prolonged aging.

Alloys↗

PubMedDate not supplied

Affinity precipitation and macroaffinity ligand facilitated three-phase partitioning for refolding and simultaneous purification of urea-denatured pectinase.

Protein refolding is an integral step in the recovery of protein activity from inclusion bodies. It is shown that affinity precipitation and macroaffinity ligand facilitated three-phase partitioning (MLFTPP) led to refolding of urea-denatured pectinase present in a commercial preparation, with simultaneous purification. Affinity precipitation consists of precipitation of the desired enzyme by complexing it with a suitable stimulus-sensitive macroaffinity ligand. This ligand in this case was alginate/esterified alginate. The complex of the polymer-pectinase could be precipitated by adding calcium ions. In MLFTPP (carried out by adding tertiary butanol and ammonium sulfate to the aqueous solution of crude enzyme and the polymer), the polymer or its complex with the enzyme form an interfacial precipitate between tert-butyl alcohol phase and aqueous phase. It is believed that in both processes, while molecular recognition of alginate/esterified alginate to pectinase facilitates their selective binding to the enzyme, the correct refolding is facilitated by preventing molecular aggregation of unfolded enzyme molecules. Three-phase partitioning with esterified alginate as the macroaffinity ligand gave 100% recovery with 4-fold purification. Affinity precipitation with 1% alginate gave 52% yield with 18-fold purification. On the other hand, use of 0.5% esterified alginate gave only 7-fold purification but with 75% recovery of activity.

Affinity Labels↗

PubMed2003

Highly dispersed green silicate and oxide pigments precipitated from model systems of postgalvanic waste.

A procedure was worked out to obtain highly dispersed green silicate and oxide pigments precipitated from postgalvanic waste. The highly dispersed chromium(III) silicates and oxides were produced from the waste, originating from chromium plating, by reduction of Cr(VI) to Cr(III) employing various reducing agents. All the reductions were conducted in an acidic medium. Solutions of Cr(III), obtained in reducing processes, were employed to precipitate silicate pigments (using sodium metasilicate solution and containing mainly chromium(III) silicates) and oxide pigments (using sodium hydroxide and containing chromium(III) oxides). The precipitated silicates and oxides were subjected to a comprehensive physicochemical analysis (estimating bulk density, capacities to absorb water, dibutyl phthalate, paraffin oil, particle size distribution, and morphology of particle surface). Precipitation process (its parameters) and heating of the reactive mixture exerts a significant effect on the principal physicochemical properties of the pigments. The heating significantly affects first of all color shade of the obtained silicate and oxide pigments as well as their dispersion. Coprecipitated chromium(III) and iron(III) silicates exhibit a brownish color and a reasonably uniform character. Apart from primary agglomerates (in the range of 414-717 nm), they contain small amounts of secondary agglomerates (in the range of 4154-6445 nm). Best physicochemical parameters have been demonstrated by chromium pigments which have been precipitated from chromium solutions reduced using hydrogen peroxide. Chromium(III) oxides deserve particular distinction since their structure includes primary particles, primary agglomerates but is completely free of secondary agglomerates. The pigments manifest a brightly green color and a low capacity to absorb water (100 cm3 x 100 g(-1)). Application of hydrophobicity-inducing agents in the course of precipitation has corrected physicochemical parameters of both the oxides and silicates of chromium(II). Their bulk densities have been clearly decreased (to as low as below 250 g x L(-1) in the case of chromium(III) silicate), while capacities to absorb paraffin oil have increased to as much as 750 cm3 x 100 g(-1) for chromium(II) oxide. The respective particle size distribution has shown a tendency for disappearance of large accumulations of secondary agglomerates.

Carcinogens, Environmental↗

PubMed2002

Precipitation scavenging of polychlorinated biphenyls and polycyclic aromatic hydrocarbons along an urban to over-water transect.

Concentrations of semi-volatile organic compounds in dissolved and particulate forms in precipitation are related to those measured concurrently in air measured at multiple locations along the tracks of several storms. In 14 paired (air and precipitation) samples collected at urban, over-water, and downwind stations around southern Lake Michigan, compound-specific total (gas + particle) precipitation scavenging ratios range from 180 to 8.2 x 10(7) (fluorene and benzo[a]pyrene, respectively) for PAHs and from 0.5 to 1.1 x 10(7) for PCBs (PCB26 and PCB194, respectively). Particle scavenging, rather than gas scavenging, is the dominant removal mechanism for both PAHs and PCBs in all samples collected along the urban to over-watertransect. Variations in measured total scavenging ratios within the 14 samples are large (from 69 to 1.00 x 10(6) for sigma-PAHs and from 290 to 88,000 for total sigma-PCBs), with larger variation between samples of differing storms collected at a single location than among samples of the same storm collected along the urban to rural transect. This minimal variance of scavenging ratio during the transport of a storm across multiple locations holds in two of three storms measured at multiple locations along the urban to over-water to rural storm track. This suggests that scavenging mechanisms are largely variable between storms and that the relative importance of each mechanism can occasionally also vary greatly during the progression of the storm from the urban to downwind locations. Furthermore, precipitation scavenging appears to be more variable between storms and generally less variable during progression of a storm down transect, indicating that meteorologic and precipitation characteristics, generally referred to here as storm type, control precipitation scavenging.

Air Pollutants↗

PubMed2002

Optimization of the isoelectric precipitation method to obtain protein isolates from amaranth (Amaranthus cruentus) seeds.

This research was conducted to evaluate the effect of extraction pH (7.8-9.2) and precipitation pH (4.3-5.7) on four selected quality attributes of protein isolates from amaranth seeds (Amaranthus cruentus) such as protein content (PC), whiteness index (WI), enthalpy of transition (EN), and denaturation temperature (DT). Ten different treatments involving extraction and precipitation pH combinations were analyzed by a central composite design; the experimental data were fitted by a second-order model using a least-squares method for each one of the four dependent variables. Response surface methodology was used for the optimization process; in addition, a common optimum value for the four dependent variables was obtained utilizing the desirability method. A confirmatory test showed that the generated regression equations could adequately predict performance of this isoelectric precipitation method. The results indicate that extraction pH and precipitation pH showed an important effect on PC, WI, and EN. However, the different combinations did not significantly affect the DT. Values of 9.2 and 8.0 for extraction pH and 5.7 for precipitation pH produced the best overall result for all responses. Finally, the results have shown that it is possible to obtain protein isolates from A. cruentus seeds at optimized values of extraction pH and precipitation pH, which presented a high protein content and good physicochemical properties.

Amaranthus↗

PubMed1996

Precipitation of proteins in supercritical carbon dioxide.

Supercritical CO2 was used as an antisolvent to form protein particles that exhibited minimal loss of activity upon reconstitution. Organic protein solutions were sprayed under a variety of operating conditions into the supercritical fluid, causing precipitation of dry, microparticulate (1-5 microns) protein powders. Three proteins were studied: trypsin, lysozyme, and insulin. Amide I band Raman spectra were used to estimate the alpha-helix and beta-sheet structural contents of native and precipitate powders of each protein. Analysis of the Raman spectral revealed minimal (lysozyme), intermediate (trypsin), and appreciable (insulin) changes in secondary structure with respect to the commercial starting materials. The perturbations in secondary structure suggest that the most significant event during supercritical fluid-induced precipitation involved the formation of beta-sheet structures with concomitant decreases of alpha-helix. Amide I band Raman and Fourier-transform infrared (FTIR) spectra indicate that higher operating temperatures and pressures lead to more extensive beta-sheet-mediated intermolecular interactions in the precipitates. Raman and FTIR spectra of redissolved precipitates are similar to those of aqueous commercial proteins, indicating that conformational changes were reversible upon reconstitution. These results suggest that protein precipitation in supercritical fluids can be used to form particles suitable for controlled release, direct aerosol delivery to the lungs, and long-term storage at ambient conditions.

Carbon Dioxide↗