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At least 235 records · Page 13Linked to original sources
PubMed1991

New modified polymeric electrodes selective to local anaesthetic compounds.

New polymeric electrodes responding to the cationic forms of tetracaine (TC), lidocaine (LD), and procaine (PC) were constructed by incorporating their ion-pair complexes (the salts of TC, LD and PC with phosphotungstic acid) into ethylene-vinyl acetate (E/VAC) copolymer. Other ion pairing agents investigated were silicotungstate and tetraphenylborate. The phosphotungstic acid resulted in the best linear and Nernstian response. A 1:1 (v/v) mixture of dioctyl phthalate (DOP) and nitrobenzene (NB) was used as plasticizer. The electrodes exhibited linear response over the concentration ranges 10(-2)-5.6 x 10(-6), 10(-2)-2.5 x 10(-5) and 10(-2)-1.8 x 10(-5) M of TC, LD and PC, respectively. pH did not affect the electrode performances within the ranges 2.7-6.3, 2.6-6.7 and 2.8-7.5 for the three electrodes, respectively. Interferences are negligible for many organic base and alkali metal cations. Cations of similar structure interfere with LD and PC, but not appreciably with TC. Direct potentiometry was used to determine these compounds in pharmaceutical preparations with accurate results.

Electrodes↗
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↗
PubMed1978

Membranes in lupin root nodules. I. The role of Golgi bodies in the biogenesis of infection threads and peribacteroid membranes.

The process of infection of lupin nodule cells by rhizobia was examined using thin-section and freeze-fracture electron-microscopic techniques to characterize the properties of different membranes and to establish relationships between them. The membranes of the Golgi bodies and the endoplasmic reticulum stained with zinc iodide-osmium tetroxide but not with phosphotungstic acid or silver. By contrast the infection thread membranes, peribacteroid membranes, plasma membranes and membranes of cytoplasmic vesicles did not stain with zinc iodide-osmium tetroxide but stained with phosphotungstic acid and silver. The peribacteroid membranes and plasma membranes are, however, different from each other since the particle density on the E face of freeze-fracture replicas of plasma membranes was twice that on the E face of the peribacteroid membranes. An examination of the tips of the infection threads in the cytoplasm of the plant cells, showed that the rhizobia bud off from the infection threads enclosed in the infection thread membranes. The rhizobia continue to divide still surrounded by membranes of plant origin, namely the peribacteroid membranes. Cytoplasmic vesicles are observed in both thin-section and freeze-fracture preparations of nodule tissue closely associated with, and apparently produced by, Golgi bodies. Formation of the walls and membranes of the infection threads and of the peribacteroid membranes involves fusion of the cytoplasmic vesicles with these membranes. It is proposed that the process of infection of plant cells in lupin nodules involves a change in the function of the Golgi body system for the biogenesis of plant cell walls and plasma membranes to include the synthesis of the walls and membranes of the infection threads and also the peribacteroid membranes.

Cell Membrane↗
PubMed2006

Ultrasound assisted phase-transfer catalytic epoxidation of 1,7-octadiene - a kinetic study.

An ultrasound assisted phase-transfer catalyzed epoxidation of 1,7-octadiene is greatly enhanced by using a cocatalyst of phosphotungstic acid in the presence of hydrogen peroxide in an organic solvent/aqueous solution two-phase medium. An active intermediate of the catalyst (Q3PW12(O)nO40, where Q = R4N+) produced from the reaction of phosphotungstic acid, hydrogen peroxide, and Aliquat 336. A rational mechanism of epoxidation is proposed to account for the reaction from the experimental evidence. The organic-phase reactions, including two series reactions, are the rate-controlling steps to produce two products, viz., 1,2-epoxy-7-octene and 1,2,7,8-diepoxyoctane. The kinetics of epoxidation, including the characteristics of the catalyst and the effect of the amount of cocatalyst, agitation speed, quaternary ammonium salts, amount of Aliquat 336, amount of hydrogen peroxide, amount of chloroform, pH value, organic solvents, and temperature on the conversion of 1,7-octadiene were investigated in detail. A kinetic model was built, from which a pseudo-first-order rate law is sufficient to describe the behavior of the reaction.

Catalysis↗
PubMed1983

[Determination of HDL-cholesterol].

For the clinical practice methods of the determination of HDL-cholesterol made their way which are based on the precipitation of apolipoprotein-B-containing lipoproteins and a determination of cholesterol following. The expensive methods of the ultracentrifugation serve as reference methods. The most-spread precipitation techniques (heparin/MCl2, dextran sulphate/CaCl2 or MgCl2 photungstic acid/MgCl2) are comparatively observed with regard to their effectiveness, practicability and methodical and technical conditions (influence of the concentration of the precipitation reagents, pH-value, temperature, incubation and centrifugation conditions). Results of own investigations as well as data from literature are presented to the problem of the harmonization of the cholesterol determination with the precipitation technique. According to the opinion of the authors for the enzymatic determination of cholesterol by means of the CHOD-PAP-method the phosphotungstic acid precipitation well stood the test, whereas for the chemical determination of cholesterol after Liebermann-Burchard in manual or automatized works the precipitation by means of dextran sulphate/CaCl2 (40 g/l, 2.0 mol/l) is to be recommended. The superabundant precipitations with phosphotungstic acid and dextran sulphate/MgCl2 (20 g/l, 2.0 mol/l) achieve higher results in Liebermann-Burchard's reaction likely on account of interferences.

Catalase↗
PubMed2000

[Changes in isolated ciliary muscle caused by repeated instillation of carbachol ointment in rabbits and effect of topically applied amlexanox].

PURPOSE: We investigated changes in ciliary muscle caused by continual contraction in rabbits and evaluated the efficacy of topically applied amlexanox, which relaxes the ciliary muscle, on such changes. SUBJECTS AND METHODS: After topical application of carbachol ointment 5 times daily for 2 weeks, the contractile responses of isolated ciliary muscles to carbachol were measured isometrically, and the ciliary smooth muscle fibers were stained with phosphotungstic acid and hematoxylin and observed histologically. 1% amlexanox solution was instilled 5 minutes before every instillation of carbachol ointment. RESULTS: Repeated topical carbachol ointment caused decreases in both contractile responses of isolated ciliary muscles to carbachol and number of ciliary smooth muscle fibers stained by phosphotungstic acid and hematoxylin. Amlexanox inhibited these changes. CONCLUSION: We found that continual contraction of the ciliary muscle caused functional and histological changes in it. These changes are thought to occur in some diseases which cause excessive contraction of the ciliary muscle. Topical amlexanox might be useful for these diseases.

Administration, Topical↗
PubMed1980

Stacking in lipid vesicle-tubulin mixtures is an artifact of negative staining.

Multilamellar stacking seen in negatively stained lipid vesicle-tubulin mixtures has been attributed to lipid-protein interactions (Caron, J. M., and R. D. Berlin, 1979, J. Cell Biol. 81:665-671). We show that this stacking is produced by the phosphotungstic acid used for staining, independent of the presence of tubulin in the sample. The morphology of negatively stained single bilayer vesicles obtained from dimyristoyl phosphatidylcholine or egg lecithin is specifically dependent upon the choice of metal stain. Uranyl oxalate maintains the appearance of unilamellar vesicles. After staining with sodium tungstate, the lipids form a network of multilayered lamellae with a periodicity of approximately 55 A. Phosphotungstic acid produces stacks of flattened vesicles with a period of approximately 115 A as well as broader multilamellar structures having a 55 A repeat. The stain-determined morphology is not markedly altered by sample concentration, incubation time, or temperature, or by the presence of tubulin.

Lipoproteins↗
PubMed1978

Ultrastructural localization of basic proteins in Trypanosoma cruzi.

The postformalin ammoniacal silver (AS) and the ethanolic phosphotungstic acid (EPTA) techniques were applied in epimastigote and trypomastigote forms of the pathogenic protozoa Trypanosoma cruzi to detect basic proteins at the ultrastructural level. With both techniques, reaction was observed in the nucleus and in some cytoplasmic vacuoles. In the kinetoplast of epimastigotes, reaction was observed only at its periphery. In trypomastigotes, however, an intense reaction was observed in the spherical kinetoplast. With the ethanolic phosphotungstic acid technique, reaction was also observed in ribosomes and at the peripheral doublet microtubules of the flagellum. The filaments which form the paraflagellar structure did not react.

Cell Nucleus↗
PubMed1982

Cytochemical studies of uterine and trophoblastic surface coats during blastocyst attachment in the ewe.

A glycoprotein coat was demonstrated on the outer surface of both the uterine and trophoblastic cells using ruthenium red, cationized ferritin, concanavalin A-peroxidase and phosphotungstic acid in HCl. No changes were observed on the uterine epithelial surface of cyclic or pregnant animals before or during blastocyst attachment (Day 15). However, the cytochemical reactions were different on the trophoblastic cells of blastocysts at Days 13 and 15, the ruthenium red and cationized ferritin sites of reaction and the concanavalin A receptors being more homogeneously distributed on the outer surface of Day-15 trophoblast. The phosphotungstic acid staining demonstrated a glycoprotein substance between the trophoblast and the uterine epithelium in adhesion areas by Day 18. The results suggest that biochemical changes occur in the composition or distribution of the trophoblastic cell coat during the process of blastocyst attachment in the ewe.

Animals↗
PubMed1993

Interindividual variation in the partition of lipoprotein(a) into lipoprotein subfractions.

In order to evaluate cardiovascular risk, we analyzed the lipid composition of HDL and the presence of lipoprotein(a) [Lp(a)] by both agarose gel electrophoresis and enzyme-linked immunoassay (ELISA). In 681 plasmas we found a close correspondence between the existence of a visible sinking pre-beta lipoprotein band and a concentration of Lp(a) higher than 300 mg/L. In the sinking pre-beta(+) samples, the HDL-cholesterol level obtained by differential ultracentrifugation was significantly higher than that obtained by precipitation with the MgCl2-phosphotungstic acid reagent; and the difference between these HDL-cholesterol values was linearly correlated with plasma Lp(a) concentration. Moreover, the other HDL lipid components and the lipid mass ratios of HDL isolated by ultracentrifugation were significantly different from those of HDL isolated by precipitation, and these changes were also correlated with plasma Lp(a). These differences are attributed to Lp(a) because it was detected in the 1.063-1.21 kg/L plasma fractions, whereas it was absent in the plasma supernates after precipitation with MgCl2-phosphotungstic acid. Although to a lesser extent, Lp(a) was also present in the LDL and VLDL density ranges and it directly depended on both the Lp(a) and the triglyceride plasma concentrations. The proportion of Lp(a) in HDL as related to that in LDL density fractions decreased as Lp(a) plasma levels increased, reflecting an interindividual variation of Lp(a) density species. Since 90% of our study population had detectable Lp(a) in plasma, the results reinforce the concept that the ultracentrifugation method is not equivalent to precipitation in most samples, and the contaminant effect of Lp(a) cannot be predicted because of Lp(a) partition into the different lipoprotein fractions.

Cholesterol, HDL↗
PubMed1988

[Synaptogenesis in the neural ganglia of the heart in the human embryo].

Electron microscopy was used to study synapse development in the cardiac ganglia of human fetuses ranging from 8 to 27 weeks of ovulation time. Staining with ethanolic phosphotungstic acid was used for analysis of synaptic active zones. Specialization of interneuronal links begins with the appearance of electron dense material on plasmalemmas of nerve cells in the places of simple contacts. First synapses with single synaptic vesicles and short osmiophilic zones were found in cardiac ganglia in 8-week-old fetuses. Large granular vesicles and mitochondria vesicles are formed from cisternae of agranular endoplasmic reticulum in the preterminal parts of axons and moved by axoplasmic transport to the osmiophilic zones of future synapses. Axodendritic synapses appeared earlier in the cardiac ganglia than axosomatic ones, the latter were observed from the middle of gestation. Transient neuroglial synapse-like contacts were found in the cardiac ganglia. Staining with phosphotungstic acid made it possible to distinguish the degree of synapse maturation according to active synaptic zones. The peculiarities of synaptic development in cardiac ganglia in comparison with that in the central nervous system may be accounted for by different origins of the neural tube and of neural crest and by the level of their phylogenic development.

Fetal Heart↗
PubMed1989

Comparison of ultracentrifugation and a precipitation method for high-density lipoprotein cholesterol quantitation in insulin-dependent diabetic patients.

We compared sodium phosphotungstic acid and magnesium chloride precipitation method for high-density lipoprotein (HDL) cholesterol quantitation with the ultracentrifugation method in 64 insulin-dependent diabetic patients with plasma triglyceride less than 3 mmol/l. The cholesterol content of HDL after precipitation of very-low-density lipoprotein (VLDL) and low-density lipoprotein (LDL) was 86% +/- 3% of the cholesterol content of HDL (q greater than 1.063) determined after ultracentrifugation at q = 1.063 (1.33 +/- 0.05 mmol/l vs 1.55 +/- 0.06 mmol/l; p less than 0.001). HDL cholesterol determined after precipitation closely correlated to HDL cholesterol determined after ultracentrifugation (r = 0.97; p less than 0.001). The absolute difference between the HDL cholesterol values obtained by the two methods was correlated to HDL cholesterol (ultracentrifugation) (r = 0.75; p less than 0.001), but it was not correlated to VLDL cholesterol, LDL cholesterol, triglyceride, HbA1c, blood glucose or serum albumin. LDL cholesterol calculated by use of Friedewald's formula was 108% +/- 4% of the cholesterol content of LDL (q = 1.019 to 1.063), determined after ultracentrifugation, but the calculated and the ultracentrifugally determined LDL cholesterol values were closely correlated (r = 0.98; p less than 0.001). These results suggest that during sodium phosphotungstic acid and magnesium chloride precipitation of plasma from diabetic patients, a constant fraction of HDL cholesterol is co-precipitated, resulting in a systematic difference in HDL cholesterol quantitation when compared with the ultracentrifugation method.

Adolescent↗
PubMed1990

Crystalline layers and three-dimensional structure of Staphylococcus aureus alpha-toxin.

Interaction of the pore-forming protein alpha-toxin from Staphylococcus aureus with lipid components from platelet membranes induces crystal formation of the toxin oligomers. Structure analysis of crystalline areas in either sodium phosphotungstic acid or a sodium phosphotungstic acid/glucose mixture has been performed with electron microscopy and image processing. Ordered domains extending up to a few micrometers were observed, particularly after application of alpha-toxin to pre-formed lipid layers. The crystals, showing tetragonal symmetry, formed either separate two-dimensional sheets or three-dimensional piles of layers. The corresponding unit cell parameter of the single layer was a = b = 109.4 A (standard deviation 2.1 A, n = 21). Incubation of the toxin with intact membranes or extracted lipids as well as application of the lipid layer technique resulted in congruous crystalline properties. The projected averaged alpha-toxin oligomer shows cyclic symmetry with a stain-filled space in the centre. The bulk of the three-dimensional model consists of four asymmetric protein units forming a ring. In addition, a small domain covers the central cavity at the face of the protein opposite to the underlying lipid. The conditions under which the tetragonal arrays are formed on the lipid layers suggest that the alpha-toxin molecule is in a conformation binding to a hydrophobic surface rather than fully inserted into a lipid bilayer.

Bacterial Toxins↗
PubMed1988

Further observations on the fine structure of tip links between stereocilia of the guinea pig cochlea.

Stereocilia of the guinea pig organ of Corti were examined by transmission electron microscopy, after fixation in glutaraldehyde and tannic acid, and postfixation and en bloc staining in osmium tetroxide, tannic acid, uranyl acetate, and phosphotungstic acid. Tip links were observed between the stereocilia. The links emerged from the tips of the shorter stereocilia in the hair bundle, running nearly at right angles to the cuticular plate, to join the side-wall of the adjacent taller stereocilium of the next row. The tip links had a fine filamentous core, approximately 6 nm in diameter. The core was surrounded by positively-staining amorphous material, which had a variable appearance from link to link. The central filament inserted into membrane specialisations at both its upper and lower ends. The results suggest that tip links have two components, and that the central filament, which has the same diameter as an actin filament, is suitable for transmitting stimulus-induced movements to the transducer channels of the stereocilium. The central filament would therefore concentrate the stimulus-induced forces onto a small area of cell membrane.

Animals↗