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Optimization of a comprehensive two-dimensional normal-phase and reversed-phase liquid chromatography system.

The present investigation is based on the evaluation of the performance of a comprehensive two-dimensional liquid chromatography (LCxLC) system during method optimization. The LCxLC set-up, operated in normal phase (NP) mode (adsorption) in the first dimension (1D) and reversed-phase (RP) mode in the second dimension (2D), is equipped with a 1D microbore silica column and a 2D monolithic C(18) column with a 10-port two position valve as the interface. A photodiode array detector is used after the 2D separation. A possible cause of peak distorsion because of the immiscibility of the mobile phases employed in the two dimensions is resolved. The optimization of the analytical run time and flow rate for both dimensions and the initial gradient in the 2D is carried out with various standard compounds. The potential and versatility of this LCxLC approach is demonstrated through the separation of 11 standard components, most of them allergens. The latter, which are characterized by a scattered distribution on the 2D space plane, underwent separation on both a hydrophobicity and polarity basis.

Chromatography, Liquid↗

NPY opposes PACAP phase shifts via receptors different from those involved in NPY phase shifts.

The suprachiasmatic nuclei (SCN) of the hamster can be maintained for several days in vitro, allowing electrophysiological investigation of the mammalian circadian clock. Application of pituitary adenylate cyclase activating peptide (PACAP) at Zeitgeber time (ZT) 6 on the first day in vitro can phase shift the rhythm of firing rate expressed by SCN neurons on a subsequent day in vitro. Here we report that co-application of neuropeptide Y (NPY) will block the phase-shifting action of PACAP. This blocking action is mimicked by [Leu31,Pro34]NPY and [D-Trp32]NPY but not by NPY(22-36) or avian pancreatic polypeptide. The results indicate that NPY has actions in the SCN via receptors distinct from the Y2 receptor, which mediates the phase-shifting action of NPY.

Animals↗

Radiation stimulation during the early stationary growth phase in Synechococcus lividus and its correlation with photooxidative stress occurring before the stationary phase.

The effects of chronic gamma radiation at dose rates ranging from 0.058 mGy d-1 on growth rate calculated during the early stationary phase were studied. A stimulatory effect occurred for all doses and for all phases of the cells selected for use in the inoculation of the medium. During the same period, the rate of nucleic acid synthesis was increased in irradiated cultures compared to control cultures. The stimulating effect always occurred in cultures irradiated from the inoculation to the eighteenth day only. This result led us to conclude that the stimulation mechanism depended upon the events occurring at the end of the exponential phase in the deceleration period. Studies on cell metabolism showed that cells presented features of photooxidative stress in this period. Increases in superoxide dismutase, glutathione reductase and glucose-6-phosphate dehydrogenase were observed in irradiated cultures. It was assumed that irradiation at very low doses could help cells to better defend against photooxidative stress by increasing oxidants that activate the glucose metabolism and C5-sugars production and nucleic acid synthesis.

Cyanobacteria↗

Distribution and determinants of the difference between diastolic phase 4 and phase 5 blood pressure.

Auscultatory blood pressure measurements from 18 325 male British civil servants were used to examine the relationship between diastolic phase 4 (D4) and phase 5 (D5). The two measures are highly correlated (r = 0.98) and this is unaffected by age. The distribution of the difference between the two indices is skewed with a median difference of 1 mmHg. Phase 4 and D5 were recorded at the same level in 31% of subjects. The difference was 5 mmHg or more in 9% of subjects, and 10 mmHg or more in 1%. The difference between the two diastolic indices depends partly on the level of blood pressure and body mass index.

Adult↗

Orientational phase transitions in the hexagonal phase of a diblock copolymer melt under shear flow

We generalize the earlier theory by Fredrickson [J. Rheol. 38, 1045 (1994)] to study the orientational behavior of the hexagonal phase of diblock copolymer melt subjected to steady shear flow. We use symmetry arguments to show that the orientational ordering in the hexagonal phase is a much weaker effect than in the lamellae. We predict the parallel orientation to be stable at low and the perpendicular orientation at high shear rates. Our analysis reproduces the experimental results by Tepe et al. [Macromolecules 28, 3008 (1995)] and explains the difficulties in experimental observation of the different orientations in the hexagonal phase.

Journal Article↗

Homogeneous nucleation of a noncritical phase near a continuous phase transition.

Homogeneous nucleation of a new phase near a second, continuous, transition, is considered. The continuous transition is in the metastable region associated with the first-order phase transition, one of whose coexisting phases is nucleating. Mean-field calculations show that as the continuous transition is approached, the size of the nucleus varies as the response function of the order parameter of the continuous transition. This response function diverges at the continuous transition, as does the temperature derivative of the free-energy barrier to nucleation. This rapid drop of the barrier as the continuous transition is approached means that the continuous transition acts to reduce the barrier to nucleation at the first-order transition. This may be useful in the crystallization of globular proteins.

Journal Article↗

Photopyroelectric ac calorimetric study of the nematic-smectic-A phase-transition line in binary liquid crystal mixtures with injected smectic-A phases.

Using a recently developed photopyroelectric ac calorimetric technique we investigate two binary liquid crystal mixtures with so-called injected smectic-A phases. Characteristic of these systems is the occurrence of nematic-to-smectic-A transition lines in the phase diagram of mixtures of pure compounds having only a nematic phase. The two binary systems are pentylcyanobiphenyl with either 4-n-propylcyclohexyl-carboxylate or 4-n-pentylphenyl 4(')-n-pentyloxybenzoate. Both these systems have domelike smectic-A ranges with narrow nematic ranges at the top. Near the top the N-SmA transitions are of first order and are crossing over to second order at a tricritical point on either side of the top with the increasing width of the nematic range. The obtained critical exponents are almost completely Fisher renormalized because of the strong concentration dependence of the nematic-to-smectic-A transition temperatures.

Journal Article↗

Phase diagram of carbon dioxide: evidence for a new associated phase.

The stability of CO(2) phases has been investigated up to 50 GPa and 750 K by in situ Raman spectroscopy and visual observations using externally heated diamond-anvil cells. A new phase (CO(2)-II) exists above 20 GPa and 500 K, which can be quenched to ambient temperature. The vibrational spectrum of this new CO(2) polymorph suggests the dimeric pairing of molecules. Based on the present in situ data and previous laser-heating results, we present new constraints for the phase diagram of carbon dioxide to 50 GPa and 2000 K. We find that carbon dioxide exhibits dramatic changes, both in the molecular configuration and in the nature of intermolecular interaction at high pressures and temperatures.

Journal Article↗

Semisharp phase field method for quantitative phase change simulations.

The standard phase field model for simulation of phase change requires an asymptotic analysis in a vanishing interface width, in order to connect the model parameters to the sharp interface parameters, which has hampered the quantitative usefulness of the method. In this Letter the method is simplified to the point that the relevant reduced problem can be solved analytically, allowing the sharp and phase field parameters to be identified, in principle, without restrictions on the model parameters. The scheme is tested for standard cases of two-dimensional solidification, showing excellent agreement with sharp interface kinetics.

Journal Article↗

Evidence of a first-order phase transition between Wigner-crystal and bubble phases of 2D electrons in higher Landau Levels.

For filling factors nu in the range between 4.16 and 4.28, we simultaneously detect two resonances in the real diagonal microwave conductivity of a two-dimensional electron system (2DES) at low temperature T approximately 35 mK. We attribute the resonance to Wigner-crystal and Bubble phases of the 2DES in higher Landau Levels. For nu below and above this range, only single resonances are observed. The coexistence of both phases is taken as evidence of a first-order phase transition. We estimate the transition point as nu=4.22.

Journal Article↗

Jahn-Teller solitons, structural phase transitions, and phase separation.

It is demonstrated that under common conditions a molecular solid subject to Jahn-Teller interactions supports stable Q-ball-like nontopological solitons. Such solitons represent a localized lump of excess electric charge in periodic motion accompanied by a time-dependent shape distortion of a set of adjacent molecules. The motion of the distortion can correspond to a true rotation or to a pseudorotation about the symmetric shape configuration. These solitons are stable for Jahn-Teller coupling strengths below a critical value; however, as the Jahn-Teller coupling approaches this critical value, the size of the soliton diverges signaling an incipient structural phase transition. The soliton phase mimics features commonly attributed to phase separation in complex solids.

Journal Article↗

Chiral nematic phase of suspensions of rodlike viruses: left-handed phase helicity from a right-handed molecular helix.

We report a study on charged, filamentous virus called M13, whose suspensions in water exhibit a chiral nematic (cholesteric) phase. In spite of the right-handed helicity of the virus, a left-handed phase helicity is found, with a cholesteric pitch which increases with temperature and ionic strength. Several sources of chirality can be devised in the system, ranging from the subnanometer to the micrometer length scale. Here an explanation is proposed for the microscopic origin of the cholesteric organization, which arises from the helical arrangement of coat proteins on the virus surface. The phase organization is explained as the result of the competition between contributions of opposite handedness, deriving from best packing of viral particles and electrostatic interparticle repulsions. This hypothesis is supported by calculations based on a coarse-grained representation of the virus.

Bacteriophage M13↗

K2[HCr2AsO10]: redetermination of phase II and the predicted structure of phase I.

Our prediction that phase II of dipotassium hydrogen chromatoarsenate, K(2)[HCr(2)AsO(10)], is ferroelectric, based on the analysis of the atomic coordinates by Averbuch-Pouchot, Durif & Guitel [Acta Cryst. (1978), B34, 3725-3727], led to an independent redetermination of the structure using two separate crystals. The resulting improved accuracy allows the inference that the H atom is located in the hydrogen bonds of length 2.555 (5) angstroms which form between the terminal O atoms of shared AsO(3)OH tetrahedra in adjacent HCr(2)AsO(10)(2-) ions. The largest atomic displacement of 0.586 angstroms between phase II and the predicted paraelectric phase I is by these two O atoms. The H atoms form helices of radius approximately 0.60 A about the 3(1) or 3(2) axes. Normal probability analysis reveals systematic error in seven or more of the earlier atomic coordinates.

Journal Article↗

Improving multiple isomorphous replacement phasing by heavy-atom refinement using solvent-flattened phases.

Solvent flattening of macromolecular MIR electron density maps is frequently used to improve the quality of the phases and the interpretability of resultant electron density maps. A new method is presented by which the heavy-atom parameters of isomorphous derivatives are refined against these same solvent-flattened phases and is shown to enhance convergence of the parameters by decoupling heavy-atom-parameter adjustment from parent-phase calculation. This approach is described here in the first example of its application in the solution of the glutaminyl-tRNA synthetase-tRNA(Gln)-ATP co-crystal structure.

Adenosine Triphosphate↗

Evidence of rapid and slow progression of cells through G2 phase in mouse epidermis: a comparison between phase durations measured by different methods.

Percentage labelled mitosis (PLM) measurements were initiated at four different times during a 24-hr period and continued for 24 hr in hairless mouse epidermis. Estimates of G2 and S phase durations (mean TG2 and mean TS) were calculated. A significant number of labelled mitoses (10--20%) was seen after 30 min in all four PLM measurements and the estimated mean TG2 varied from 1.4 to 2.5 hr and was in agreement with values from PLM measurements in other epithelial tissues. These mean TG2 values were much shorter than expected from [3H]TdR double labelling experiments and from a multiparameter cell kinetic study in hairless mouse epidermis and did not reflect the circadian variations seen in these studies. The differences in estimates of phase durations can be explained by postulating two G2 cell populations; one with a rapid and another with a slow rate of cell cycle progression. The cells with the higher rate are mainly registered by the PLM method, whereas those with the lower rate largely escape detection by this method. TG2 estimates from PLM measurements in mouse epidermis therefore do not reflect the phase duration of the entire G2 population. It is also concluded that circadian variations in TS can not be accurately registered by the PLM method.

Animals↗

Three phase elimination of oxytetracycline in veal calves; the presence of an extended terminal elimination phase.

The pharmacokinetics of oxytetracycline were studied after both intravenous (i.v.) and intramuscular (i.m.) administration to a group of five veal calves. Blood samples were taken frequently during the terminal elimination phase in order to calculate a reliable elimination rate constant. Because of the low limit of quantification of the method of analysis used, oxytetracycline plasma concentrations could be monitored over a 12-day period of time. After the intravenous administration of oxytetracycline, data were fitted according a three-compartment model. After i.m. administration, plasma-concentration-time curves could best be described by a two-compartment model. It was demonstrated that a very slow terminal elimination phase was present both after i.v. and i.m. administration with a half-life of approximately 95 h. The data show that this phase cannot be explained by slow absorption from the injection site and that release of oxytetracycline incorporated into bone is not a likely explanation.

Absorption↗

Metabolism of apigenin by rat liver phase I and phase ii enzymes and by isolated perfused rat liver.

The metabolism of apigenin, a low estrogenic flavonoid phytochemical, was investigated in rat using liver models both in vitro (subcellular fractions) and ex vivo (isolated perfused liver). In vitro, phase I metabolism led to the formation of three monohydroxylated derivatives: luteolin which was the major metabolite (K(m) = 22.5 +/- 1.5 microM; V(max) = 5.605 +/- 0.090 nmol/min/mg protein, means +/- S.E.M.), scutellarein, and iso-scutellarein. These oxidative pathways were mediated by cytochrome P450 monooxygenases (P450s). The use of P450 inhibitors and inducers showed that CYP1A1, CYP2B, and CYP2E1 are involved. In vitro studies of phase II metabolism indicated that apigenin underwent conjugation giving three monoglucuronoconjugates and one monosulfoconjugate. Luteolin led to the formation of four monoglucuronoconjugates, two sulfoconjugates, and one methylconjugate identified as diosmetin. Ex vivo during the apigenin perfusion of an isolated rat liver, none of the phase I metabolites could be recovered. In contrast, two monoglucuronoconjugates and one of the sulfoconjugates of apigenin already identified in vitro were recovered. Moreover, two new derivatives were isolated and identified as a diglucuronoconjugate and a glucuronosulfoconjugate. This work provides new data about the metabolism of apigenin and shows the interest value of using various experimental models in metabolic studies.

Animals↗

Macroscopic separation of dense fluid phase and liquid phase of phosphorus.

Structural transformation between a dense molecular fluid and a polymeric liquid of phosphorus that occurred at about 1 gigapascal and 1000 degrees C was investigated by in situ x-ray radiography. When the low-pressure fluid was compressed, dark and round objects appeared in the radiograph. X-ray diffraction measurements confirmed that these objects were the highpressure liquid. The drops grew and eventually filled the sample space. Decompressing caused the reverse process. The macroscopic phase separation supported the existence of a first-order phase transition between two stable disordered phases besides the liquid-gas transition. X-ray absorption measurements revealed that the change in density at the transition corresponds to about 40% of the density of the high-pressure liquid.

Journal Article↗