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At least 145 records · Page 8Linked to original sources
PubMed1991

Stereospecific assignment of beta-methylene protons in larger proteins using 3D 15N-separated Hartmann-Hahn and 13C-separated rotating frame Overhauser spectroscopy.

3J alpha beta coupling constants and complementary nuclear Overhauser data on the intraresidue C alpha H-C beta H distances form an essential part of the data needed to obtain stereospecific assignments of beta-methylene protons in proteins. In this paper we show that information regarding the magnitude of the 3J alpha beta coupling constants can be extracted from a semi-quantitative interpretation of relative peak intensities in a 3D 15N-separated 1H-1H Hartmann-Hahn 1H-15N multiple quantum coherence (HOHAHA-HMQC) spectrum. In addition, we demonstrate that reliable information on the intraresidue C alpha H-C beta H distances, free of systematic errors arising from spin diffusion, can be obtained from a 3D 13C-separated 1H-1H rotating frame Overhauser effect 1H-13C multiple quantum coherence (ROESY-HMQC) spectrum. The applicability of these experiments to larger proteins is illustrated with respect to interleukin-1 beta, a protein of 153 residues and 17.4 kDa molecular weight.

Amino Acids↗
PubMed1990

Production and separation of peptides from proteins stained with Coomassie brilliant blue R-250 after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Proteins stained with Coomassie brilliant blue on polyacrylamide gels were digested with lysylendopeptidase in the presence of sodium dodecyl sulfate. Peptide production was similar to that under ordinary conditions of digestion. Peptides were recovered easily and efficiently from the gel pieces and separated by HPLC. The present method for preparation of peptides from proteins separated by sodium dodecyl sulfate gel electrophoresis is quite simple and can be used for sequence analysis of proteins in general at the subnanomolar level.

Amino Acid Sequence↗
PubMed1991

Phase separation temperatures of mixtures of Triton X-114 and Triton X-45: application to protein separation.

Triton X-114 solutions separate above 22 degrees C into two immiscible aqueous phases. The more dense phase is enriched in detergent, and the less dense phase is depleted of detergent, relative to the original single phase. This phenomenon has been used to partition proteins according to hydrophobicity. The phase separation temperature is sensitive to the length of the polyoxyethylene headgroup. When Triton X-45, with a shorter headgroup, is mixed with Triton X-114 in various proportions, the phase transition temperature can be adjusted anywhere between 0 and 22 degrees C. Partitioning properties of the resulting mixtures are similar to those of Triton X-114 alone.

1-Phosphatidylinositol 4-Kinase↗
PubMed1993

Expression is computed separately from facial identity, and it is computed separately for moving and static faces: neuropsychological evidence.

We report data contrasting the processing of facial identity from static photographs, and facial expression from static and moving images, in two patients with face processing impairments. One patient is markedly impaired at perceiving facial identity and he is poor at discriminating facial expression and gender from static photographs of faces. In contrast, he performs normally when required to make judgements of facial expression and gender to faces depicted by sets of moving light dots. The second patient is relatively good at perceiving facial identity, but poor at judging emotional expression from both static and moving faces. The data are consistent with the existence of separate processes for encoding face identity and facial expression, and, furthermore, indicate the separate encoding of expression from moving and static images.

Cerebrovascular Disorders↗
PubMed1983

Separation of subtypes of depression using discriminant analysis. Separation of bipolar endogenous depression from nonendogenous ("Neurotic") depression.

We derived a discriminant function separating patients with bipolar endogenous depression ("melancholia") from patients with nonendogenous ("neurotic") depression, and showed that the difference between the groups was not one of overall severity of illness alone. The discriminant function (DF) included 5 clinical items. We reduced the DF to a discriminant index (DI) with integral item weights, and trichotomized the DI scores into two definite classifications and an intermediate, uncertain classification. We cross-validated this DI in a separate group of patients, and found no decrease in the accuracy of classification on cross-validation. Thirty-three of 41 (80%) of the patients in the cross-validation group were classified by the DI; 26 of 33 (79%) correctly. We also validated the DI classification against an external, biological marker, the dexamethasone suppression test (DST). The DI predicted the DST result with the same accuracy as the clinical diagnoses did, supporting the validity of the DI.

Adult↗
PubMed1993

Sequence of centromere separation. Minor satellite DNA does not influence separation of inactive centromeres in transformed cells of mouse.

Neoplastic cells may carry inactive centromeres on some multicentric, yet stable, chromosomes. We report that some inactive centromeres in L929 mouse cells do not contain minor satellite DNA, the DNA fraction which has been suggested to constitute the centromere. We compared the sequence of separation of inactive centromeres carrying the minor satellite with those lacking this fraction. The sequence of separation appears to be independent of whether or not the inactive centromeres carry the minor satellite DNA. The timing of replication of the inactive centromeres is also independent of this DNA. Hence, minor satellite of mouse is not a factor in holding together the subunits of inactive centromeres. Extension of these results to active centromeres might suggest that the minor satellite DNA is not a factor responsible for adhesion of the two centromere sub-units up until late meta-anaphase.

Animals↗
PubMed1990

Separate hemoglobin standards for blacks and whites: a critical review of the case for separate and unequal hemoglobin standards.

There have been a number of reports that suggest mean differences between the hemoglobin (Hgb) levels of black and white Americans. Many of those reports have had as their basis various national US nutrition surveys. Early reports indicated that blacks had a 1.0 g/dl lower mean Hgb than whites. Later reports put the mean Hgb difference between 0.5 and 0.73 g/dl. Recent reports, in which either iron intake or iron status have been measured and controlled, indicate a mean Hgb difference of between 0.2 and 0.3 g/dl. Despite the variability in these mean Hgb differences, some reports call for the establishment of separate Hgb standards for black Americans. The need for separate Hgb standards is critically reviewed in light of alternative interpretations of early data and newer knowledge about the development, detection, and significance of iron deficiency. The suitability of national cross-sectional survey data to answer questions about Hgb standards without adequate experimental studies in questioned, especially since no genetic studies have been performed to indicate genetic differences in Hgb levels. Policy implications of a change from current standards are also discussed.

Adolescent↗
PubMed2006

Nanofiltration for the separation of pharmaceuticals from nutrients in source-separated urine.

The potential of nanofiltration for the separation of pharmaceutical and estrogenic compounds from salts in urine was investigated with the aim of producing a micropollutant-free nutrient solution that can be used as a fertilizer. A fresh urine solution and a synthetic solution of similar inorganic composition were tested at different pH values in order to investigate their separation behavior. These solutions were spiked with the micropollutants propranolol, ethinylestradiol, ibuprofen, diclofenac and carbamazepine. Among the membranes tested, NF270 showed the best performance with respect to the retention of micropollutants. The optimum retention of micropollutants was obtained at values of around pH 5. At this point, the retention of all micropollutants in non-hydrolysed urine was above 92%, while the corresponding value for the synthetic urine solution was above 73%. From the results, it can be concluded that the retention mechanism is determined by steric and electrostatic effects as well as by the partitioning of the micropollutants in the membrane. The nutrients urea and ammonia were well permeated, but phosphate and sulfate were almost completely retained. Nanofiltration can consequently be used to produce a permeate which contains most of the nitrogen and a greatly reduced proportion of micropollutants.

Filtration↗
PubMed1998

Separation of metalloprotein complexes in serum by size-exclusion chromatography. Optimisation of the separation parameters retention behaviour and recovery employing radiotracers.

A suitable procedure was developed for speciation analysis of metalloprotein complexes in serum using directly coupled size-exclusion chromatography and an element-specific detector. Two column matrices used for size-exclusion chromatography (TSK G 3000 SW and Asahipak GS 520) were investigated with respect to the recovery and retention behaviour for metalloprotein complexes. Optimisation of the separation parameters (buffer type, concentration, pH) was achieved by means of metalloprotein complexes marked with radiotracers. For speciation of serum the matrix in the Asahipak GS column is more efficient. Given optimal eluent characteristics (100 mM Tris, pH 7.4) the recovery of the elements investigated (sodium, calcium, iron and zinc) was 100%. Further, the retention behaviour (retention time, ratios of the peak areas) remained unchanged for several successive separations.

Buffers↗
PubMed1998

Separation of microsomal cytochrome b5 via phase separation in a mixed solution of Triton X-114 and charged dextran.

The successful introduction of a charged dextran into the Triton X-114 phase separation system for the selective extraction of cytochrome b5 (cyt. b5) in liver microsomes is described. In the absence of charged dextran, 55% of total microsomal proteins and 84% of cyt. b5 were extracted into the surfactant-rich phase. In the presence of anionic dextran sulfate, the extractability of total microsomal proteins was greatly reduced while that of cyt. b5 was increased. After triplicate extraction, cyt. b5 was purified more than 10-fold from microsomes with a recovery of 91% in the surfactant-rich phase. In view of its operational simplicity, this method provides a good means for the partial purification of cyt. b5 prior to chromatographic separations.

Animals↗
PubMed2005

Separation of peptides from myoglobin enzymatic digests by RPLC. Influence of the mobile-phase composition and the pressure on the retention and separation.

The influence of the mobile-phase composition and the pressure on the chromatographic separation of the peptides from the enzymatic digest of myoglobin was studied under linear conditions. The retention behavior of these tryptic peptides was measured under isocratic conditions with different mobile-phase compositions, ranging from 9 to 28% (v/v) acetonitrile in 0.1% (v/v) aqueous trifluoroacetic acid. The effect of the pressure was studied by analyzing the separation of the tryptic peptides under different average column pressures between 14 and 220 bar, at 13, 20, and 26% (v/v) acetonitrile. The differences between the partial molar volumes of these peptides in the stationary and mobile phases were derived from these results. All the measurements were performed on a 10-cm-long C18-bonded, end-capped monolithic column. The results obtained illustrate the highly complicated behavior of the complex peptide mixtures afforded by tryptic digestion. The capacity factors of the analyzed peptides do not depend linearly on the acetonitrile concentration but follow exactly a quadratic relationship. The adsorption changes of partial molar volumes are in good agreement with other literature data. The consequences of the influence of the average column pressure (hence of the flow rate) on the column phase ratio and on the retention factors of the peptides are discussed. The retention pattern of the complex mixture is affected by both the mobile-phase composition and the pressure, and the resolution of certain peptide pairs is so much affected by the pressure that inversions in the elution order of some pairs are observed.

Adsorption↗
PubMed2006

Monolithic column HPLC separation of intact proteins analyzed by LC-MALDI using on-plate digestion: An approach to integrate protein separation and identification.

A method is developed to integrate a protein separation by monolithic capillary reversed-phase high-performance liquid chromatography to on-probe tryptic digestion for subsequent analyses by MALDI-TOF MS and MALDI-TOF/TOF MS. The method provides a means of directly interfacing separations to MALDI-MS, reducing the amount of time required for traditional procedures involving in-solution enzymatic digestion and sample cleanup prior to MALDI-MS analysis. When used with pI-based fractionation as a first dimension, it provides a means of analyzing complex mixtures of proteins with minimal sample handling and cleanup. The use of monolithic capillary columns sufficiently resolved intact proteins so that peptide mass fingerprinting analysis by MALDI-TOF MS resulted in the identification of close to 40 unique proteins from 120 ng of sample obtained from a prefractionated MCF10 cell line at pH 6.34, where the identifications of several of these proteins were also confirmed by intact MW and tandem mass spectrometric analysis. The reproducibility of this method has been demonstrated to be sufficient for the purpose of protein identifications. Experimental values of protein intact MW are obtained and compared to that expected for each protein identified.

Amino Acid Sequence↗
PubMed1997

Improved chiral separations using a polymerized dipeptide anionic chiral surfactant in electrokinetic chromatography: separations of basic, acidic, and neutral racemates.

Two polymeric chiral anionic surfactants [poly(sodium N-undecylenyl-L-valine) (poly-L-SUV) and poly(sodium N-undecylenyl-L-valine-valine) (poly-L-SUVV)] are compared as pseudostationary phases for chiral separations of basic, acidic, and neutral enantiomers. Parameters such as pH, concentration and type of background electrolyte, concentration of polymerized chiral surfactants, and injection size were studied to investigate the migration behavior and optimize the chiral resolution of several racemic analytes. At equivalent monomer concentrations, the migration factors for cationic enantiomers were larger with poly-L-SUV than with poly-L-SUVV. In contrast, the reverse was true for anionic enantiomers. However, in both cases, chiral recognition was significantly enhanced with poly-L-SUVV as compared to that with poly-L-SUV. It is interesting to note that the separation selectivity and resolution of a neutral racemate were slightly better with the latter, but only at the expense of longer analysis time and lower efficiencies.

Buffers↗
PubMed1988

High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 2. Effect of pulse time and electric field strength and implications for models of the separation process.

Bacteriophage DNAs annealed into linear oligomeric concatemers were used to examine the quantitative pulsed-field gel electrophoretic behavior of different-sized DNAs as a function of electrical field strength and pulse time. Three zones of resolution are observed for increasingly larger DNAs. In the first two zones, the electrophoretic mobility decreases linearly with increasing DNA size. The separation in zone 2 is roughly twice that in zone 1. The largest DNA molecules do not resolve at all and migrate in a compression zone. Mobility in zone 1 increases linearly with the electric field strength and decreases with the inverse of the pulse time. The behavior of DNA in zone 2 is qualitatively similar. However, the effect of field strength and pulse time on the separations in each zone is quite different. The results for zone 1 are generally consistent with the predictions of several existing physical models of pulsed-field gel electrophoresis, but no model accounts for all of the observed behavior in the three zones.

Bacteriophages↗
PubMed1996

Perceptual separability, decisional separability, and the identification-speeded classification relationship.

Three observers participated in several sessions of identification and speeded classification. Predictions from F. G. Ashby and W. T. Maddox's (1994a) response time (RT) theory of speeded classification were tested. A key theoretical concept is decisional separability (DS), that is, that the decision about the level of 1 component does not depend on the perceived value of the other component. The theory predicted that DS would hold in the filtering task but would be violated in the redundancy task, resulting in the redundancy gains. To summarize the results, (a) DS held in the filtering conditions, but not in the redundancy conditions; (b) redundancy gains occurred; (c) despite the redundancy gains, strong evidence was obtained that the stimulus components are perceptually separable; (d) 2 new models that each assumed RT decreases with the distance between the percept and the decision bound provided good accounts of the RT distributions and accuracy rates; and (e) the shift from identification to speeded classification influenced both perceptual and decisional processes.

Adult↗
PubMed1983

Response tempo and separable--integral responding: evidence for an integral-to-separable processing sequence in visual perception.

Three experiments were conducted to examine the relation between response tempo and separable-integral responding. A restricted classification paradigm, in which similarity-based (integral) and dimensionally based (separable) classifications were pitted against one another, was used with the dimensions of length and density (all experiments) and size and brightness (Experiment 3). The subjects were college students (Experiments 1 and 2) and kindergarten, second-, and fifth-grade children (Experiment 3). In all three experiments, rapid responding was associated with fewer dimensional and more similarity responses than was slow responding. This result held when response tempo was simply measured (Experiments 1 and 3) and when it was manipulated by the experimenter (Experiment 2). The results were interpreted to be consistent with models of dimensional processing in which holistic, integral processing precedes analytic dimensional processing.

Child↗
PubMed2003

Preserving informational separability and violating decisional separability in facial perception and recognition.

The holistic encoding hypothesis (M. J. Farah, K. D. Wilson, M. Drain, & J. N. Tanaka, 1998) proposes that faces are encoded and used in perception and cognition as relatively undifferentiated wholes. A previous study (M. J. Wenger & E. M. Ingvalson, 2002) found very little support for the strong version of this hypothesis and instead found evidence that shifts in decisional criteria may be important. This study provides a replication and stronger test of those findings, demonstrating consistent violations of decisional separability and preservation of informational separability in both immediate perception and delayed recognition.

Adult↗
PubMed1971

The separation of different cell classes from lymphoid organs. IV. The separation of lymphocytes from phagocytes on glass bead columns, and its effect on subpopulations of lymphocytes and antibody-forming cells.

Four separate effects can be demonstrated when lymphoid cell suspensions are passed through columns of siliconed glass beads. (a) A temperature-dependent "active adherence" of phagocytic cells, such as macrophages and polymorphs. (b) A temperature-independent and selective trapping by "physical adherence" of particular classes of lymphoid cells, including certain antibody-forming cells. (c) A "size-filtration" effect that traps larger cells, but only becomes significant with beads below 100 micro in diameter. (d) A selective retention of damaged cells, which occurs with all columns under all conditions tested. An active adherence column technique has been developed to separate phagocytes from lymphocytes while minimizing selection within the lymphocyte population by physical adherence or size filtration. In less than 10 min at 37 degrees C it reproducibly produces a preparation of mouse spleen lymphocytes >500-fold depleted of active macrophages, and approximately 50-fold depleted of active polymorphs, with good over-all cell recoveries and cell viability. The lymphocyte fraction appears fully active in its ability to initiate immune responses to at least two different antigens, but is changed in over-all composition and selectively depleted in certain classes of antibody-forming cells.

Animals↗