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At least 613 records · Page 34Linked to original sources
PubMed2005

Changes in uterine protein secretion during luteal and follicular phases and detection of phosphatases during luteal phase of estrous cycle in buffaloes (Bubalus bubalis).

Changes in uterine proteins during different reproductive states and their functional significance though known in other species have not been established in buffaloes. An attempt has been made to unravel the changes in composition of buffalo uterine secretion with growth and regression of corpora-lutea during early, mid and late luteal and follicular phase of estrous cycle using gel filtration and electrophoresis techniques. Also the phosphatases activities in luteal phase uterine secretions have been studied. Gel filtration chromatography analysis revealed a protein peak in void volume of the column, the intensity of which was more in all the luteal phase samples than follicular phase samples. Alkaline phosphatase was also found eluted in the void volume. The other three uterus-specific peaks (Peaks V-VII) were detected below 13.7 kd molecular weight. There were at least five peaks of acid phosphatases activity in chromatogram. Silver staining of SDS-PAGE gel detected as many as 40 protein bands in the uterine fluid of which nine proteins were glycoproteins. Molecular weight (MW) comparison revealed the major protein band at 66 kd which could be serum albumin. Comparison of uterine proteins with serum protein bands revealed a 93.5 kd glycoprotein in buffalo serum that did not appear in uterine fluid and at least 11 uterus-specific protein bands (506, 470, 241, 114, 49, 38, 33, 26, 19.2, 16, and 14.3 kd). The 38 and 19.2 kd bands were luteal-stage specific. Intense periodic acid Schiff's (PAS) stained bands in uterine proteins compared to serum indicated glycosylation process in endometrial epithelial cells. The study suggested that buffalo uterine secretion contained mainly serum and several uterus-specific proteins of which few were luteal phase specific. Further study on characterizing the unique or most abundant proteins and defining their role in uterine functions would help to address the cause of low reproduction rate in buffaloes.

Acid Phosphatase↗
PubMed1986

Inverted micellar intermediates and the transitions between lamellar, cubic, and inverted hexagonal lipid phases. I. Mechanism of the L alpha----HII phase transitions.

A model for the thermotropic transitions between lamellar (L alpha) and inverted hexagonal (HII) phases is developed. According to this model, the first structures to form during the L alpha----HII transition are inverted micellar intermediates (IMI). The structure, formation rates, and half-lives of IMI ("lipidic particles") were described previously. IMI coalesce in the planes between apposed bilayers to form two types of HII phase precursors. The first is a monolayer-encapsulated HII tube (RMI), which forms via coalescence of IMI in pearl-string fashion. These structures have been proposed previously based on electron microscopic evidence. I show that if only RMI form, L alpha in equilibrium HII transitions cannot occur on observed time scales (faster than seconds). I propose that a second type of intermediate, a line defect (LD), forms as well. LD should form via IMI-IMI coalescence in significant numbers, and elongate rapidly into structures consisting of two apposed halves of HII tubes. Transitions via LD can occur in less than seconds, the time depending on the fraction of IMI-IMI coalescence events producing LD and the number of IMI per unit of bilayer area. Hysteresis in the phase transition temperature may be due to the difference in water content of the two phases and their low water permeabilities. The model is in qualitative agreement with morphological, NMR, and x-ray diffraction data on phospholipid systems. The results are relevant to IMI-mediated interactions between unilamellar bilayer vesicles, and to the structure of inverted cubic phases observed in some phospholipid systems. These will be discussed in subsequent publications (D. P. Siegel, manuscript in preparation).

Colloids↗
PubMed1996

Topology of gel-phase domains and lipid mixing properties in phase-separated two-component phosphatidylcholine bilayers.

The influence of the lipid mixing properties on the lateral organization in a two-component, two-phase phosphatidylcholine bilayer was investigated using both an experimental (fluorescence recovery after photobleaching (FRAP)) and a simulated (Monte Carlo) approach. With the FRAP technique, we have examined binary mixtures of 1-stearoyl-2-capryl-phosphatidylcholine/1,2-distearoyl-phosphat idylcholine (C18C10PC/DSPC), and 1-stearoyl-2-capryl-phosphatidylcholine/1,2-dipalmitoyl-phospha tid ylcholine (C18C10PC/DPPC). Comparison with the 1,2-dimyristoyl-phosphatidylcholine/1,2-distearoyl-phosphatidylcholine (DMPC/DSPC) previously investigated by FRAP by Vaz and co-workers (Biophys. J., 1989, 56:869-876) shows that the gel phase domains become more effective in restricting the diffusion coefficient when the ideality of the mixture increases (i.e., in the order C18C10PC/DSPC-->C18C10PC/DPPC-->DMPC/DSPC). However, an increased lipid miscibility is accompanied by an increasing compositional dependence: the higher the proportion of the high-temperature melting component, the less efficient the gel phase is in compartmentalizing the diffusion plane, a trend that is best accounted for by a variation of the gel phase domain shape rather than size. Computer-simulated fluorescence recoveries obtained in a matrix obstructed with obstacle aggregates of various fractal dimension demonstrate that: 1) for a given obstacle size and area fraction, the relative diffusion coefficient increases linearly with the obstacle fractal dimension and 2) aggregates with a lower fractal dimension are more efficient in compartmentalizing the diffusion plane. Comparison of the simulated with the experimental mobile fractions strongly suggests that the fractal dimension of the gel phase domains increases with the proportion of high-temperature melting component in DMPC/DSPC and (slightly) in C18C10PC/DPPC.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2000

Interpretive optimisation strategy applied to the isocratic separation of phenols by reversed-phase liquid chromatography with acetonitrile-water and methanol-water mobile phases.

An optimisation protocol is presented for the resolution of complex mixtures in isocratic RPLC with binary mobile phases of organic solvent and water, which is based on the prediction of peak position and shape of the individual compounds. A good description of the retention was achieved through the application of statistical weights to the widely used linear or quadratic relationships between the logarithm of the retention factor (log k) and the organic solvent concentration in the mobile phase. The maximisation of the product of peak purities for each compound is shown as a competitive resolution strategy versus the worst value of a selectivity parameter. Peak purities allow one to associate a single resolution value to each compound, which is not affected by the identity of the interfering peaks. It is shown how when full resolution is not achieved with a single mobile phase, the same experimental data set (retention factors, asymmetries and efficiencies) can be used for finding two or three optimal complementary mobile phases (CMPs). Each CMP resolves fully some compounds in the mixture, while the remaining compounds can overlap among them. The elementary limiting resolutions, which measure the maximal separation degree for each compound, are also given as a useful guide in the selection of the elution conditions. A mixture of 13 phenols (phenol, chloro-, bromo-, nitro- and methyl-derivatives), eluted with acetonitrile-water or methanol-water mobile phases, is used to show the proposed methodology.

Acetonitriles↗
PubMed1988

Effect of column temperature on the retention of peropyrene-type polycyclic aromatic hydrocarbons on various chemically bonded stationary phases in reversed-phase liquid chromatography.

The effect of column temperature on the reversed-phase retention of polycyclic aromatic hydrocarbons has been investigated using various chemically bonded phases. Four solutes, coronene, tetrabenzo[a,cd,j,lm]perylene, tetrabenzo[a,cd,f,lm]perylene and benzo[lm]phenanthro[4,5,6-abcd]perylene, were used as the test probes. The temperature dependences of the retention are almost linear (the logarithm of capacity factor is proportional to the reciprocal of the column absolute temperature) with monomeric C18, monomeric C18 with endcapping and diphenyl bonded phases, while non-linear behaviour was observed with polymeric C18 phases. These differences in behaviour of the stationary phases are interpreted in terms of their structural differences.

Chromatography, Liquid↗
PubMed2001

Laterally attached liquid crystalline polymers as stationary phases in reversed-phase high-performance liquid chromatography. V. Study of retention mechanism using linear solvation energy relationships.

A linear solvation energy relationship model was used to characterize the retention behavior of a stationary phase based upon a nematic side-on liquid crystalline polymer (SOLCP) in reversed-phase liquid chromatography. The set of solutes was constituted of a high variety of compounds whose molecular sizes were considerably smaller than the mesogenic unit size. The results showed good statistical fits for these retention data in 65:35, 75:25 and 85:15 (v/v) methanol-water mobile phases. Both the cavity term and excess molar refraction are the most important favorable retention-governing parameters, whereas the solute hydrogen bond acceptor basicity is the most unfavorable retention parameter. Hydrophobicity and pi-pi interactions decrease strongly when the percentage of methanol increases, leading to an important retention decrease despite the fact that the hydrogen bond interaction weakens as the organic solvent is added. The shape recognition ability of this side-on liquid crystalline stationary phase on polycyclic aromatic hydrocarbon solutes is partly explained by the solutes' high polarizability due to the presence of pi-electrons. However, the solute polarizability is not sufficient and a stationary phase's "structure effect" must to be taken into account for the shape discrimination observed. The strong interaction between liquid crystal molecules caused likely a adsorption retention mechanism rather than a partition mechanism.

Chromatography, High Pressure Liquid↗
PubMed1987

Evaluation of advanced silica packings for the separation of biopolymers by high-performance liquid chromatography. IV. Mobile phase and surface-mediated effects on recovery of native proteins in gradient elution on non-porous, monodisperse 1.5-microns reversed-phase silicas.

The reversed-phase chromatography of proteins by gradient elution with acidic, low-ionic-strength aqueous-organic eluents is often associated with losses of the biological activity of the protein. In this study, the enzymatic activities of catalase, horseradish peroxidase and pepsin were examined under static and dynamic column conditions on non-porous, monodisperse 1.5-microns reversed-phase silicas with various n-alkyl ligands. Catalase readily lost its enzymatic activity under the influence of the acidic aqueous-organic eluents in the absence of the reversed-phase packing, whereas peroxidase was partially deactivated as a result of combined mobile phase and stationary phase effects but regained its activity on storage after elution. The enzymatic activity of pepsin was found to be very dependent on the column residence time and on the type of bonded n-alkyl ligand employed.

Catalase↗
PubMed1991

Alternative mobile phases for the reversed-phase high-performance liquid chromatography of peptides and proteins.

The use of a high content of acetic acid as mobile phase additive for the reversed-phase high-performance liquid chromatography (RP-HPLC) of several proteins and extracts of biological tissues was evaluated for a divinylbenzene (DVB)-based stationary phase, and the separations obtained with acetic acid gradients in acetonitrile, isopropanol or water were compared with classical polypeptide RP-HPLC on silica C4 with trifluoroacetic acid (TFA)-acetonitrile. The separation patterns for recombinant derived interleukin-1 beta (IL-1 beta) on the C4 column eluted with TFA-acetonitrile and the DVB column eluted with acetic acid-acetonitrile were similar, but only the polymeric column was able to separate the components present in an iodinated IL-1 beta preparation. Neither eluent had any harmful effect on the biological activity of IL-1 beta isolated after RP-HPLC. Several standard proteins could be separated when the polymeric column was eluted with acetic acid gradients in acetonitrile, isopropanol or water and, although the separation efficiency with acetic acid in water was lower than that in combination with classical organic modifiers, insulin, glucagon and human growth hormone (hGH) were eluted as sharp, symmetrical peaks. The recoveries of insulin and hGH were comparable for all three mobile phases (80-90%). The separation patterns obtained from a crude acetic acid extract of a normal and a diabetic, human pancreas analysed using acetic acid gradients with or without organic modifiers were found to be similar and comparable to those obtained on a silica C4 column eluted with an acetonitrile gradient in TFA. The principal differences resulted from the use of different UV wavelengths (215 nm for TFA-acetonitrile, 280 nm for acetic acid). Acetic acid extracts of recombinant derived hGH-producing Escherichia coli were separated on the DVB column eluted with an acetic acid gradient in water. Although the starting material was a highly complex mixture, the hGH isolated after this single-step purification was surprisingly pure (as judged by sodium dodecyl sulphate-polyacrylamide gel electrophoresis). Consequently several (pure) polypeptides and complex biological samples were separated on a polymeric stationary phase eluted with acetic acid gradients in water without the use of organic modifiers.

Animals↗
PubMed1987

Resolution of enantiomeric amides on a cellulose tribenzoate chiral stationary phase. Mobile phase modifier effects on retention and stereo-selectivity.

The effect of the steric structure and concentration of the mobile phase modifier on the retention (kappa') and stereoselectivity (alpha) of a series of enantiomeric amides has been investigated. The amides were chromatographed on a commercially available cellulose tribenzoate chiral stationary phase (CSP) using mobile phases composed of hexane and two homologous series of alcohols: methanol, ethanol, 1-propanol and 2-propanol, 2-butanol, 2-pentanol, 2-hexanol. The results of the study indicate that the alcoholic mobile phase modifiers compete with the solutes for achiral and chiral binding sites and that the steric bulk around the hydroxyl moiety of the modifier plays a role in this competition. Increased steric bulk tends to result in increased kappa' and alpha. However, the results also suggest that the effect of the alcoholic mobile phase modifiers on stereoselectivity may also be due to binding to achiral sites near or at the chiral cavities of the CSP which alters the steric environment of these cavities.

Alcohols↗
PubMed1987

Elution behaviour of peropyrene-type polycyclic aromatic hydrocarbons in various chemically bonded stationary phases in reversed-phase liquid chromatography.

The elution behaviour of polycyclic aromatic hydrocarbons (PAHs, 13 of the 16 U.S. Environmental Protection Agency priority pollutants and peropyrene types) was studied on several chemically bonded stationary phases (octadecylsilicas, di- and triphenylsilicas, naphthylethylsilica and pyrenylethylsilica) under reversed-phase conditions. The results showed that the elution order of peropyrene-type PAHs is highly dependent on the degree of planarity of the solute and on the orderlines of the bonded phases, whereas no definite differences were found in the retention behaviour of the 13 small PAHs on various stationary phases. The characteristics of all the stationary phases could be classified by statistical cluster analysis.

Chemical Phenomena↗
PubMed1986

Protein conformational effects in hydrophobic interaction chromatography. Retention characterization and the role of mobile phase additives and stationary phase hydrophobicity.

We have studied the conformational behavior of alpha-lactalbumin (alpha-LACT) in hydrophobic interaction chromatography (HIC). Retention characterization in terms of Z (slope of plot of ln k' vs. ln phi B, where k' is the capacity factor and phi B is the volume fraction of mobile phase B) has been explored, and the relationship of Z to other slopes, such as S (slope of the plot of ln k' vs. phi B) has been derived. The reasons for the sensitivity of Z to conformational change are discussed. The enhanced broadening of alpha-LACT in a temperature transition region of conformational change has been studied by spectral analysis using on-line photodiode array detection. The influence of Ca2+ and Mg2+ addition to the mobile phase is further explored. Since alpha-LACT is a calcium binding protein, addition of this metal leads to stabilization, i.e. higher column temperatures are required for conformational change. On the other hand, addition of Mg2+ appears to destabilize the protein. We have explored the use of a more hydrophobic support, C2-(ethyl) ether phase, for the elution of alpha-LACT. In this case, two widely separated peaks are observed. By spectral analysis the first peak is shown to be native and the later eluted, broad second peak to be an unfolded mixture of species. As previously observed in reversed-phase liquid chromatography, the second peak grows at the expense of the first, as the column temperature is raised. The second peak also grows as the contact time of the protein with the surface increases. This behavior can be ascribed to the conformational change of alpha-LACT in the column, the late eluted species under the second peak binding significantly more strongly to the phase than the native peak. Reinjection of the late eluted fraction reveals that reformation of the native species takes place in solution within 30 min. As before, addition of Ca2+ reduces the extent of unfolding under any specific condition. These results add further to our understanding and ability to control conformational changes in high-performance liquid chromatography.

Animals↗
PubMed2002

Characterization and comparison of the chromatographic performance of conventional, polar-embedded, and polar-endcapped reversed-phase liquid chromatography stationary phases.

We have evaluated and compared the performance of several conventional C18 phases with those possessing either a polar-endcapping group or a polar-embedded group within the primary alkyl ligand and found distinct differences in the chromatographic behavior among the three groups, as well as a high degree of variability within each group. The trend is for the polar-endcapped phases to display similar hydrophobic retention characteristics as the conventional C18 columns, but to express higher hydrogen bonding capacities and silanol activity. The polar-embedded phases displayed the opposite behavior, with a greatly reduced hydrophobic nature compared to the conventional and polar-endcapped C18 phases, and also a very much reduced silanol activity. Most interestingly, it appears that ionic or dipole interactions play a significant role in the overall retention behavior of the polar-embedded phases towards basic and acidic analytes.

Chromatography, High Pressure Liquid↗
PubMed2002

Characterisation of reversed-phase stationary phases for the liquid chromatographic analysis of basic pharmaceuticals by thermodynamic data.

This paper describes the characterisation of reversed-phase liquid chromatography (RPLC) columns using thermodynamic measurements. Retention versus 1/T data were used to construct Van't Hoff plots. The slope of these plots indicates the standard enthalpy of transfer of the analyte from the mobile to the stationary phase. The standard entropy can be calculated from the intercept. Van't Hoff plots were linear for the investigated RPLC columns, meaning that for basic analytes over the temperature range studied no changes in the retention mechanism occurred. Enthalpies and entropies of transfer of basic analytes from the mobile to the stationary phase revealed information about the types of interaction of protonated and neutral compounds with the stationary phases. However, a clear view using the present set of basic compounds on how these thermodynamic data may explain the observed substantial differences in peak symmetry cannot be given. It is considered that addition of N,N-dimethyloctylamine (DMOA) to the eluent will results in a dynamically coating of the stationary phase. Addition of DMOA to the eluent resulted for protonated basic compounds in a reduction of both enthalpy and entropy. In practice, with DMOA in the eluent symmetrical peaks were obtained. It is assumed that this is due to blocking residual silanols and/or ion exclusion effects.

Chromatography, Liquid↗
PubMed1999

Chromatographic properties of reversed-phase stationary phases under pressure- and electro-driven conditions.

Seven different reversed-phase (RP) stationary phases were examined under high-performance liquid chromatographic (pressure-driven, HPLC), and capillary electrochromatographic (electro-driven, CEC) conditions. Characterization of the stationary phases was performed following well-established test procedures providing a number of distinct column descriptors: hydrophobicity, hydrophobic selectivity and silanol activity. These parameters were used to describe the behavior of the RP-columns under both HPLC and CEC conditions. It is shown that chromatographic characteristics of porous RP-phases greatly depend on the mode of operation. By contrast, column descriptors of a non-porous viz. solid RP-phase material hardly differed for HPLC and CEC conditions.

Chromatography, High Pressure Liquid↗
PubMed2000

Influence of mobile phase composition on electroosmotic flow velocity, solute retention and column efficiency in open-tubular reversed-phase capillary electrochromatography.

The effects of some experimental parameters, such as the volume fraction and type of organic modifier in the mobile phase, and the concentration, type and pH of the buffer on the electroosmotic flow velocity, the retention behavior of test solutes, and the column efficiency have been investigated in capillary electrochromatography (CEC) using an open-tubular column of 9.60 microm I.D. with a porous silica layer chemically modified with C18 as stationary phase. The retention of a group of polycyclic aromatic hydrocarbons (PAHs) used as a test mixture varied significantly by changing the organic modifier content in the hydroorganic mobile phase according to the reversed-phase-like selectivity of the stationary phase. In addition, an increase in the percentage of organic modifier resulted in a slight increase in the linear velocity of the EOF. On the other hand, when the phosphate buffer concentration was increased over the range 1-50 mM, the electroosmotic mobility fell dramatically, the retention of the solutes decreased steadily, and the plate height showed a significant increase. The results obtained with phosphate, trishydroxymethylaminomethane or 2-morpholinoethanesulfonic acid as buffers were similar when pH remained constant. Optimization in CEC was essential to achieve further enhancement of separation performance, because the analysis time and separation resolution are essentially affected when varying operating parameters. Separations of seven PAHs with more than 100000 plates are presented within 4 min analysis time.

Buffers↗
PubMed1988

Intermittent Wolff-Parkinson-White syndrome due to phase 3 and phase 4 block: disappearance of rapid ventricular response during atrial fibrillation.

A 65-year-old man with Wolff-Parkinson-White syndrome type A had a rapid ventricular response over an accessory pathway during atrial fibrillation. Four months later, electrophysiological study revealed the following properties: 1) The antegrade accessory pathway conduction showed phase 3 and phase 4 block. 2) The retrograde concealed accessory pathway conduction eliminated phase 4 block in the antegrade accessory pathway conduction. 3) Premature ventricular excitation arose from the accessory pathway or from the ventricular muscle close to its distal end. 4) Atrial fibrillation during isoproterenol infusion did not show rapid ventricular response. Spontaneous impairment of accessory pathway conduction due to phase 3 and phase 4 block was suspected.

Aged↗
PubMed1999

Effect of liquid inhibitors on PCDD/F formation. Prediction of particle-phase PCDD/F concentrations using PLS modelling with gas-phase chlorophenol concentrations as independent variables.

Emissions of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) from municipal waste incineration are currently a subject of considerable public concern because of their extreme toxicity. PCDD/F formation in incineration processes is being studied widely, but little work has been done on their inhibition. We studied the effect of two liquid inhibitors, sodium ammonium hydrogen phosphate (NAHF) and urea (H2NCONH2), on PCDD/F formation in the combustion of liquid fuel doped with copper and chlorine using a pilot-scale plant. The inhibitors were injected into the flue gas stream at a temperature of 725 degrees C, whereupon both the chlorophenol and PCDD/F concentrations decreased. Particle-phase PCDD/F concentrations in particular decreased by up to 90% with NAHF and 70% with urea, but gas phase reduction took place only with urea. The results suggest that the formation of PCDD/Fs is hindered in the particle phase at the early stages of the PCDD/F formation chain, probably even before precursors such as chlorophenols have been formed. As a consequence, particle-phase PCDD/F concentrations can be predicted by a PLS (partial least-squares) approach with the gas-phase chlorophenol concentrations as independent variables. The structure and partial charges of Cu(+)-urea complex were calculated by the HF/3-21G basis set.

Benzofurans↗
PubMed2002

A new approach based on monitoring of phase formation kinetics for examination of biological particles and cells, using aqueous two phase polymer systems.

An original technique of use of two-phase polymer systems as an analytical research method is described. The technique is based on the absorbance change of two-phase systems in visible spectrum during formation of the phases. Dynamics of this process was demonstrated as the kinetic curves. Addition of studied objects (macromolecules or cells) to the two-phase system modified the shape of the kinetic curve, depending on their individual surface properties. The technique has the following advantages as compared with traditional procedures of the particle surface analysis with the help of two-phase polymer systems: examination of particles with partition coefficients approaching zero; multiple analyses of the same samples; use of interphase as a matrix for study of spontaneous formation of studied particle complexes. The opportunities of the technique were demonstrated in a series of previous authors' works.

Actins↗