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LC-MS compatible HPLC separation for xenobiotics and their phase I and phase II metabolites: simultaneous anion exchange and reversed-phase chromatography.

Benzene and five of its mammalian metabolites, phenol, phenyl-beta-D-glucuronide, phenylsulfate, phenylmercapturate, and t,t-muconic acid, are separated on a methylstyrene-divinylbenzene-based anion exchange--reversed-phase column. The retention of the model compounds is manipulated by modifying the type, ionic strength, and pH of the mobile phase buffer, and the type and percent of organic mobile phase modifier. The separations developed are compatible with both particle beam (PB) and thermospray (TSP) interfaces used in liquid chromatography-mass spectrometry (LC-MS). These separations should be adaptable to the study of a wide range of xenobiotics and their phase I and phase II metabolites, and provide an LC-MS compatible alternative to the use of iron pairing reagents.

Benzene↗

Bare silica as a reverse-phase stationary phase: liquid chromatographic separation of antihistamines with buffered aqueous organic mobile phases.

Unbonded silica gel is an effective support for reverse-phase liquid chromatographic separations of lipophilic amines. Simple buffered aqueous-organic mobile phases provide rapid isocratic separations of antihistamines. Phenylpropanolamine hydrochloride, chlorpheniramine maleate, and dextromethorphan hydrobromide are separated in 8 min on bare silica with a mobile phase of 75% methanol and 25% water which is 0.01 M in (NH4)2HPO4. Quantitation is reproducible. A wide variety of additional compounds may be separated using the same mobile phase.

Chromatography, Liquid↗

Mobile phase effects in reversed-phase chromatography. VII. Dependence of retention on mobile phase composition and column temperature.

The relationship between logarithmic retention factor, kappa, in reversed-phase chromatography and operating conditions including mobile phase composition and column temperature is still controversial. Earlier, the following analytical relationship was proposed for use with alkyl benzenes: kappa = A1 phi (1 - Tc/T) + A2/T + A3 where phi is volume fraction organic co-solvent of the hydro-organic mobile phase, T is absolute temperature and A1-A3 are parameters appropriate to the eluite and stationary phase. Insofar as its use had not been tested with polar eluites that are retained by only one mechanism, its description of the retention of 54 polar and apolar eluites was examined and compared to equations with other, usually more complex, relationships between retention and operating conditions. The following four-parameter equation described best most data: kappa = A1 phi (1 - Tc/T) + A2/T + A3 + A4 phi. But neither this nor any other four-parameter equation under examination here emerged solely as the best expression for the dependence of retention on the eluent composition and temperature. The average relative errors were 7.8% and 6% with use of the three- and the four-parameter equations which were found to be best for a given eluite, respectively. In view of the small decrease in error but significant increase in complexity and in data points with the use of the four-parameter equation, the three-parameter equation is recommended for general use.

Chemical Phenomena↗

High-performance liquid chromatography determination of praziquantel enantiomers in human serum using a reversed-phase cellulose-based chiral stationary phase and disc solid-phase extraction.

A sensitive HPLC method for the quantification of praziquantel enantiomers in human serum is described. The method involves the use of a novel disc solid-phase extraction for sample clean-up prior to HPLC analysis and is also free of interference from trans-4-hydroxypraziquantel, the major metabolite of praziquantel. Chromatographic resolution of the enantiomers was performed on a reversed-phase cellulose-based chiral column (Chiralcel OJ-R) under isocratic conditions using a mobile phase consisting of 0.1 M sodium perchlorate-acetonitrile (66:34, v/v) at a flow-rate of 0.5 ml/min. Recoveries for R-(-)- and S-(+)-praziquantel enantiomers were in the range of 84-89% at 50-500 ng/ml levels. Intra-day and inter-day precisions calculated as R.S.D. were in the ranges of 3-8% and 1-8% for both enantiomers, respectively. Intra-day and inter-day accuracies calculated as percent error were in the 0.2-5% and 0.3-8% ranges for both enantiomers, respectively. Linear calibration curves were in the concentration range 10-600 ng/ml for each enantiomer in serum. The limit of quantification of each enantiomer was 10 ng/ml. The detection limit for each enantiomer in serum using a UV detector set at 210 nm was 5 ng/ml (S/N=2).

Antiplatyhelmintic Agents↗

High-performance liquid chromatographic analysis of ondansetron enantiomers in human serum using a reversed-phase cellulose-based chiral stationary phase and solid-phase extraction.

A stereospecific HPLC method was developed for the assay of R-(-)- and S-(+)-ondansetron enantiomers in human serum. The method involves the use of solid-phase extraction for sample clean-up and is also free of interference from 6-hydroxyondansetron, 7-hydroxyondansetron and 8-hydroxyondansetron, the three major metabolites of ondansetron. Chromatographic resolution of the enantiomers was performed on a reversed-phase cellulose-based chiral column (Chiralcel OD-R) under isocratic conditions using a mobile phase consisting of 0.7 M sodium perchlorate-acetonitrile (65:35, v/v) at a flow-rate of 0.5 ml/min. Recoveries at 200 ng/ml levels were more than 90% for both ondansetron enantiomers. Intra-day and inter-day precisions calculated as R.S.D.s were in the 0.3-5% and 2-8% ranges for both enantiomers, respectively. Intra-day and inter-day accuracies calculated as percent error were in the 0.3-11.5% and 0-3% ranges for both enantiomers, respectively. Linear calibration curves were obtained for each enantiomer in serum in the concentration range 15-750 ng/ml. The limit of quantitation of each enantiomer was 15 ng/ml. The detection limit for each enantiomer in serum using UV detection at 210 nm was 7 ng/ml (S/N=2).

Antiemetics↗

Resolution and quantitation of pentazocine enantiomers in human serum by reversed-phase high-performance liquid chromatography using sulfated beta-cyclodextrin as chiral mobile phase additive and solid-phase extraction.

A sensitive and stereospecific HPLC method was developed for the analysis of (-)- and (+)-pentazocine in human serum. The assay involves the use of a phenyl solid-phase extraction column for serum sample clean-up prior to HPLC analysis. Chromatographic resolution of the pentazocine enantiomers was performed on a octadecylsilane column with sulfated-beta-cyclodextrin (S-beta-CD) as the chiral mobile phase additive. The composition of the mobile phase was aqueous 10 mM potassium dihydrogenphosphate buffer pH 5.8 (adjusted with phosphoric acid)-absolute ethanol (80:20, v/v) containing 10 mM S-beta-CD at a flow-rate of 0.7 ml/min. Recoveries of (-)- and (+)-pentazocine were in the range of 91-93%. Linear calibration curves were obtained in the 20-400 ng/ml range for each enantiomer in serum. The detection limit based on S/N=3 was 15 ng/ml for each pentazocine enantiomer in serum with UV detection at 220 nm. The limit of quantitation for each enantiomer was 20 ng/ml. Precision calculated as R.S.D. and accuracy calculated as error were in the range 0.9-7.0% and 1.2-6.2%, respectively, for the (-)-enantiomer and 0.8- 7.6% and 1.2-4.6%, respectively, for the (+)-enantiomer (n=3).

Analgesics, Opioid↗

Response latency, latency-based receiver-operating characteristics, and uncertainty about interaural phase in the detection of in-phase and out-of-phase tones presented in noise.

Subjects were given the task of detecting tonal signals presented in a continuous background of white noise which was always in phase at the ears (No). The signals were either in phase (So), or phase-reversed (S pi) at the ears. The analysis of response latencies (for similar levels of performance) indicated that there were consistent differences in the processing of the two types of signals. Latencies were longer, and somewhat more variable, for S pi than for So signals. It was also found that the theory of signal detectability can contribute to an understanding of what and how decisions are made. Latencies were treated as confidence ratings in order to determine latency-based receiver operating characteristics (LROCs) for the detection task. It was observed that the LROCs for the interaural condition No-So tend to leave the origin at a steeper angle than do those for the interaural condition No-S pi, presumably reflecting the different sensory and decision processes employed in the two conditions. When the two interaural conditions were intermixed within blocks of trials, performance was slightly impaired in comparison to the situation in which the interaural condition remained constant throughout blocks of trials.

Acoustic Stimulation↗

[The indirect measurement of arterial pressure as a function of cuff width in patients in the immediate preoperative phase, the phase of entry into the operating room and in the postanesthesia phase].

The goal of this study was to compare blood pressure measures using two sizes of cuffs: one standard width (12 cm) and other with correct width that is 0.38 of arm circumference as recommended by American Heart Association. The comparisons were done among surgical patients in mediate perioperative phase, reception area of operation room and postoperative phase during the staying of the patient in the recovery room. The results demonstrated that the arterial blood pressure was hypoestimated by the use of the standard width cuff, reaching values up 30 mmHg in the systolic arterial pressure and 30 mmHg in the diastolic ones.

Adolescent↗

Detecting hepatocellular carcinoma: value of unenhanced or arterial phase CT imaging or both used in conjunction with conventional portal venous phase contrast-enhanced CT imaging.

OBJECTIVE: Because rates of detection of hypervascular neoplasms by conventional dynamic incremental-bolus CT are lower than rates of detection of hypovascular tumors by CT and because both unenhanced CT imaging and arterial phase helical CT imaging may increase the detection of hypervascular tumors, such as hepatocellular carcinoma, we evaluated the value of unenhanced and arterial phase CT imaging used in conjunction with conventional portal venous phase CT imaging in patients with hepatocellular carcinoma. MATERIALS AND METHODS: Unenhanced and biphasic helical contrast-enhanced CT studies were performed on 81 patients with proven hepatocellular carcinoma. Arterial phase and portal venous phase images were obtained at 20-50 sec and at 60-100 sec, respectively. Three blinded readers evaluated portal venous phase images for the number of liver lesions. On separate dates, the readers compared the arterial phase images with the portal venous phase images and the unenhanced images with the portal venous phase images. The readers recorded the number of lesions that were seen on portal venous phase images and that were also detected on unenhanced or arterial phase images as well as the number of additional lesions seen on unenhanced or arterial phase images. Consensus readings of unenhanced, arterial phase, and portal venous phase images were obtained in the 42 patients who had definitive surgery or follow-up CT scans, documenting the total tumor burden in this patient subgroup. RESULTS: The readers identified 286-310 lesions on portal venous phase images. On unenhanced images, the readers identified 223-244 of the lesions seen on portal venous phase images and an additional 45-55 lesions that were not seen on portal venous phase images. Arterial phase imaging revealed 245-269 of the lesions seen on portal venous phase images and an additional 89-111 lesions that were not seen on portal venous images. The diagnosis of tumor was possible only on unenhanced images in two (3%) of 81 patients and only on arterial phase images in seven patients (9%). In the subset of 42 patients with proof of tumor burden, 157 proven lesions were found. Consensus readings identified 127 (81%) of these lesions on portal venous phase images, 98 (62%) of these lesions on unenhanced images, and 120 (76%) of these lesions on arterial phase images. Of the 30 lesions not seen on portal venous phase images, nine were seen on both unenhanced and arterial phase images, three were seen on unenhanced images only, and 18 were seen on arterial phase images only. CONCLUSION: In patients with known or suspected hepatocellular carcinoma, the use of unenhanced or arterial phase images or both in addition to conventional portal venous phase images resulted in more tumors being detected. The combination of arterial phase and portal venous phase images revealed significantly more hepatocellular carcinoma lesions than did the combination of unenhanced and portal venous phase images.

Adult↗

T1-weighted spoiled gradient-echo MR imaging of focal hepatic lesion: comparison of in-phase vs opposed-phase pulse sequence.

The goal of our prospective study was to compare quantitatively and qualitatively in-phase and opposed-phase T1-weighted breath-hold spoiled gradient-recalled-echo (GRE) MR imaging technique for imaging focal hepatic lesion. Thirty-eight patients with 53 focal hepatic lesions had in-phase (TR = 12.3 ms, TE = 4.2 ms) and opposed-phase (TR = 10.1 ms, TE = 1.9 ms) GRE (flip angle = 30 degrees , bandwidth +/- 32 kHz, matrix size 256 x 128, one signal average) MR imaging at 1.5 T. Images were analyzed quantitatively by measuring the lesion-to-liver contrast and for lesion detection. In addition, images were reviewed qualitatively for lesion conspicuity. Quantitatively, lesion-to-liver contrast obtained with in-phase (3. 22 +/- 1.86) and opposed-phase pulse sequence (3.72 +/- 2.32) were not statistically different (Student's t-test). No difference in sensitivity was found between in-phase and opposed-phase pulse sequence (31 of 53, sensitivity 58 % vs 30 of 53, sensitivity 57 %, respectively). Two lesions not seen with opposed-phase imaging were detected with in-phase imaging. Conversely, one lesion not seen on in-phase imaging was detected on opposed-phase imaging so that the combination of in-phase and opposed-phase imaging yielded detection of 32 of 53 lesions (sensitivity 60 %). Qualitatively, lesion conspicuity was similar with both techniques. However, in-phase images showed better lesion conspicuity than opposed-phase images in 9 cases, and opposed-phase images showed better lesion conspicuity than in-phase images in 7 cases. No definite advantage (at a significant level) emerged between in-phase and opposed-phase spoiled GRE imaging. Because differences in lesion conspicuity and lesion detection may be observed with the two techniques in individual cases, MR evaluation of patients with focal hepatic lesion should include both in-phase and opposed-phase spoiled GRE imaging.

Adult↗

Effect of cell exposure to top or bottom phase prior to cell partitioning in dextran-poly(ethylene glycol) aqueous phase systems: erythrocytes as a model.

Cells exposed to dextran (Dx)-rich bottom phase prior to cell partitioning in Dx-poly(ethylene glycol) (PEG) aqueous two-phase systems have lower partition ratios than cells exposed to PEG-rich top phase. Aspects of this previously observed phenomenon were explored. In the present work charge-sensitive phases made with Dx T500 and PEG 8000 were used exclusively. It was found that: (1) even on countercurrent distribution (CCD) red cells (RBC) loaded in bottom phase have a lower apparent partition ratio, G, than the same cells loaded in top phase; (2) when part of the same cell population is loaded into top phase and part into bottom phase of the same load cavities for CCD, with the cells loaded into top or bottom bearing an isotopic tracer (51Cr), the cells loaded into top phase have a higher G value than the cells loaded into bottom phase; (3) the shift in the CCD curves of human or of rat RBC between cells loaded in top or bottom phase using systems having the same polymer concentration (though different salt compositions) shows no striking difference and is, for the number of experiments run, not statistically significant; (4) when the quantity of cells loaded for CCD is reduced from 10(9) to 10(8), the G value of cells loaded in top phase is reduced slightly while that of cells loaded in bottom phase is diminished more appreciably; (5) increasing polymer concentrations yield larger differences in G values between (rat) RBC loaded in top or bottom phase; (6) when cells exposed to top or bottom phase, respectively, are centrifuged and suspended in bottom or top phase, respectively, their CCD patterns are qualitatively similar to cells exposed to these latter respective phases initially; (7) rat RBC populations containing 59Fe-labeled cells of different but distinct age are fractionated on CCD irrespective of whether loaded in top or bottom phase. An exception are populations containing very young mature labeled cells (e.g., 4-d old) which are resolved when loaded in top phase but not in bottom phase. Thus cell populations exist which can be resolved by CCD when loaded in one of the phases but not when loaded in the other. Glutaraldehyde-fixed rat RBC containing 4-d old labeled cells are fractionated by CCD irrespective of whether loaded in top or bottom phase.

Animals↗

Selectivity of stationary phases in reversed-phase liquid chromatography based on the dispersion interactions.

Selectivity of 15 stationary phases was examined, either commercially available or synthesized in-house. The highest selectivity factors were observed for solute molecules having different polarizability on the 3-(pentabromobenzyloxy)propyl phase (PBB), followed by the 2-(1-pyrenyl)ethyl phase (PYE). Selectivity of fluoroalkane 4,4-di(trifluoromethyl)-5,5,6,6,7,7,7-heptafluoroheptyl (F13C9) phase is lowest among all phases for all compounds except for fluorinated ones. Aliphatic octyl (C8) and octadecyl (C18) phases demonstrated considerable selectivity, especially for alkyl compounds. While PBB showed much greater preference for compounds with high polarizability containing heavy atoms than C18 phase, F13C9 phase showed the exactly opposite tendency. These three stationary phases can offer widely different selectivity that can be utilized when one stationary phase fails to provide separation for certain mixtures. The retention and selectivity of solutes in reversed-phase liquid chromatography is related to the mobile phase and the stationary phase effects. The mobile phase effect, related to the hydrophobic cavity formation around non-polar solutes, is assumed to have a dominant effect on retention upon aliphatic stationary phases such as C8, C18. In a common mobile phase significant stationary phase effect can be attributed to dispersion interaction. Highly dispersive stationary phases such as PBB and PYE retain solutes to a significant extent by (attractive) dispersion interaction with the stationary phase ligands, especially for highly dispersive solutes containing aromatic functionality and/or heavy atoms. The contribution of dispersion interaction is shown to be much less on C18 or C8 phases and was even disadvantageous on F13C9 phase. Structural properties of stationary phases are analyzed and confirmed by means of quantitative structure-chromatographic retention (QSRR) study.

Chromatography, Liquid↗

Studies on ovarian and adrenal steroids at different phases of the menstrual cycle: II. A comparative assessment of the circadian variation in steroid and lutropin levels during the follicular, periovulatory and luteal phases.

In six normally menstruating women, ovarian and adrenal steroids and biologically active lutropin (LH) were measured in peripheral plasma samples collected every 3 h. during a period of 39 h. in the early follicular phase, periovulatory period or luteal phase of three consecutive cycles. The purpose of the study was to assess the influence of the phases of the cycle on the diurnal variation in the levels of different steroids and lutropin following the elimination of the between subject variation. Cortisol, 17-hydroxypregnenolone, dehydroepiandrosterone and androstenedione showed a marked circadian rhythm in all three phases of the cycle. No changes in the levels of cortisol, 17-hydroxypregnenolone and dehydroepiandrosterone with the phase of the cycle were observed when the "morning" samples were considered; however, when the "evening" samples were analyzed, the levels of these steroids were higher in the luteal phase than in the follicular phase. As a result of this increase, the amplitude of the circadian rhythm of these steroids considerably decreased in the luteal phase. The "morning" as well as the "evening" increase observed in the levels of androstenedione during the periovulatory period was not able to mask the circadian rhythm. A circadian rhythm in pregnenolone, 17-hydroxyprogesterone, testosterone, and dihydrotestosterone levels was detected only in certain phases of the cycle. All these steroids showed a circadian rhythm in the early follicular phase. The rhythm of pregnenolone and 17-hydroxyprogesterone was still present in the periovulatory period but was no longer detectable in the luteal phase, meanwhile that of testosterone and dihydrotestosterone was not demonstrable neither in the periovulatory period nor in the luteal phase. Compared to the levels of the follicular phase, an increase in pregnenolone and 17-hydroxyprogesterone levels was observed in the periovulatory period which was followed by a further rise in the luteal phase. This increase completely masked the circadian rhythm in the luteal phase. An inverse circadian rhythm in lutropin levels was detected during the luteal phase. The "morning" values were lower than those found during the "evening" period. No such changes were observed in the other phases of the cycle. In none of the phases studied did the levels of progesterone or estradiol show any circadian variation. The data indicate that a circadian rhythm in the peripheral levels of a given steroid mainly depends on the relative contributions of the ovaries and adrenals and that these contributions exhibit major differences at the various phases of the cycles. It is concluded that - in contradistinction to the situation in the human male - in normally menstruating women, the peripheral levels of steroids of predominantly gonadal origin do not exhibit a circadian rhythmicity.

Adrenal Glands↗

Is the second phase of a biphasic defibrillation waveform the defibrillating phase?

Why some biphasic waveforms defibrillate with lower energies than monophasic waveforms of similar duration is unknown. One hypothesis is that the first phase of a biphasic waveform acts as a conditioning, hyperpolarizing prepulse to prepare for defibrillation by a second depolarizing phase. To test whether the second phase of a biphasic waveform is the defibrillating phase, three monophasic waveforms, an ascending ramp (A), a square wave (S), and a descending ramp (D), were compared to three biphasic waveforms with A, S, or D in the first phase (biphasic first phase) and three biphasic waveforms with A, S, or D in the second phase (biphasic second phase). Two defibrillation thresholds for each waveform were performed in 18 open chest pigs and mean defibrillation thresholds were compared. In nine pigs 16-msec monophasic and 16/16-msec biphasic waveforms were ranked by mean current and energy at defibrillation threshold. The ranks were the same for monophasic and biphasic second phase waveforms: for mean current A < S = D and for energy A < S < D. The ranks were different for the biphasic first phase waveforms: for mean current S < A = D and for energy S < A = D. Although ranks for the 16-msec monophasic waveforms matched those for the 16/16-msec biphasic second phase waveforms, the biphasic waveforms had higher mean currents and energies at defibrillation threshold. In nine pigs defibrillation thresholds for 6-msec monophasic and 6/6-msec biphasic waveforms were ranked. For mean current the ranks were monophasic: A < S = D; biphasic first phase: A = S = D; and biphasic second phase: S = D < A. For energy the ranks were monophasic: A = S < D; biphasic first phase: A = S = D; and biphasic second phase: S = D < A. Thus, ranks for the 6-msec monophasic waveforms differed from those for the 6/6-msec biphasic second phase waveforms. For 16/16-msec biphasic waveforms, less effective for defibrillation than corresponding 16-msec monophasic waveforms, these results support the hypothesis that the second phase of a biphasic waveform defibrillates since the defibrillation efficacy of a 16/16-msec biphasic waveform is related to the defibrillation efficacy of its second phase waveshape. However, for clinically useful 6/6-msec biphasic waveforms, more effective for defibrillation than 6-msec monophasic waveforms, the hypothesis is not supported because the ability of a 6/6-msec biphasic waveform to defibrillate is unrelated to the defibrillation efficacy of its second phase waveshape.

Animals↗

Optimal small-capacitor biphasic waveform for external defibrillation: influence of phase-1 tilt and phase-2 voltage.

BACKGROUND: Biphasic waveforms have been reported to be more efficacious than monophasic waveforms for external defibrillation. This study examined the optimal phase-1 tilts and phase-2 leading-edge voltages with small capacitors (60 and 20 microF) for external defibrillation. We also assessed the ability of the "charge-burping" model to predict the optimal waveforms. METHODS AND RESULTS: Two groups of studies were performed. In group 1, 9 biphasic waveforms from a combination of 3 phase-1 tilt values (30%, 50%, and 70%) and 3 phase-2 leading-edge voltage values (0.5, 1.0, and 1.5 times the phase-1 leading-edge voltage, V1) were tested. Phase-2 pulse width was held constant at 3 ms in all waveforms. Two separate 60- microF capacitors were used in each phase. The energy value that would produce a 50% likelihood of successful defibrillation (E50) decreased with increasing phase-1 tilt and increased with increasing phase-2 leading-edge voltage except for the 30% phase-1 tilt waveforms. In group 2, 9 waveforms were identical to the waveforms in group 1, except for a 20- microF capacitor for phase 2. E50 decreased with increasing phase-1 tilt. Phase-2 leading-edge voltage of 1.0 to 1.5 V1 appeared to minimize E50 for phase-1 tilt of 50% and 70% but worsened E50 for phase-1 tilt of 30%. There was a significant correlation between E50 and residual membrane voltage at the end of phase 2, as calculated by the charge-burping model in both groups (group 1, R2=0.47, P<0.001; group 2, R2=0.42, P<0.001). CONCLUSIONS: The waveforms with 70% phase-1 tilt were more efficacious than those with 30% and 50%. The relationship of phase-2 leading-edge voltage to defibrillation efficacy depended on phase-2 capacitance. The charge-burping model predicted the optimal external biphasic waveform.

Animals↗

Molecular cloning of G1 phase mRNAs from a subtractive G1 phase cDNA library.

Many G1-phase-specific mRNAs have been identified from various normal or transformed cells based on serum induction and re-entry into the cell cycle from quiescence. However, these mRNAs may not represent some important genes expressed during G1 phase in continuously cycling cells. The eukaryotic cell cycle possesses two cdk (cyclin-dependent kinase) dependent regulatory gates through which cells pass during late G1 phase and G2 phase of each cycle. Subtractive hybridization was employed to synthesize a high R0t fraction cDNA library enriched in sequences expressed during G1 phase prior to passage through the G1-phase gate. To prepare G1-phase cells from continuously cycling cell populations, G1-phase HeLa cells were collected by centrifugal elutriation and highly synchronous S phase cells were obtained by double thymidine block followed by centrifugal elutriation. A G1-phase subtractive cDNA library was prepared by subtracting G1-phase cDNA with a 10-fold excess of S-phase mRNA. Single-stranded, G1-phase cDNAs were isolated by oligo(dA) chromatography. The library was screened with a high R0t fraction subtractive probe population. Following two rounds of screening, 20 positive clones were obtained. Northern blot analysis indicated that six of these clones were enhanced in expression level during G1 phase when compared with S phase. Nucleotide sequence comparison of each clone with the GenBank data base revealed that hG1.11 was highly homologous (99%) to the apoferritin light chain gene and clones hG1.6, hG1.10, hG1.17, and hG1.18 represented new G1-phase-enriched members of four human ribosomal protein gene families (71-95% homology). The last clone, hG1.1, encoded a highly charged polypeptide not previously identified. Additional study of these G1-phase-enriched mRNAs will be required to determine their role in cell cycle progression and the G1-phase gateway through which cells transit as they proceed through the cell cycle.

Actins↗

The effect of frequency on the visual perception of relative phase and phase variability of two oscillating objects.

Relative phase has been studied extensively as a measure of interlimb coordination. Only two relative phases, namely 0 degrees and 180 degrees, are stably produced at the preferred frequency (approximately 1 Hz). When frequency is increased, movement at 180 degrees becomes unstable and relative phase typically switches to 0 degrees, which remains stable at higher frequencies. The current study was designed to investigate the perception of relative phase and of phase variability. Observers viewed two circles moving rhythmically in a computer display. Mean phases varied from 0 degrees to 180 degrees in 30 degrees steps. Phase variability at each mean phase varied from 0 degrees to 5 degrees, 10 degrees, and 15 degrees phase standard deviation (SD). Frequency of oscillation was either 0.75 Hz or 1.25 Hz. One group of ten observers judged mean relative phase. Another group judged phase variability. As predicted, increase in frequency yielded an increase in perceived phase variability at 180 degrees mean phase and other mean phases, but not at 0 degrees mean phase. In contrast, increase in actual phase variability affected judgments of 0 degrees mean phase most strongly. A second control experiment showed that the frequency effects were not produced by changes in display durations or frames per cycle of oscillation. The results are consistent with those in studies of interlimb coordination and indicate that understanding of interlimb coordination requires further investigation of phase perception.

Adolescent↗

Effect of electrostatic interactions on phase stability of cubic phases of membranes of monoolein/dioleoylphosphatidic acid mixtures.

To elucidate effects of electrostatic interactions resulting from surface charges on structures and phase stability of cubic phases of lipid membranes, membranes of 1-monoolein (MO) and dioleoylphosphatidic acid (DOPA) (DOPA/MO membrane) mixtures have been investigated by small-angle x-ray scattering method. As increasing DOPA concentration in the DOPA/MO membrane at 30 wt% lipid concentration, a phase transition from Q(224) to Q(229) phase occurred at 0.6 mol% DOPA, and at and above 25 mol% DOPA, DOPA/MO membranes were in the L(alpha) phase. As NaCl concentration in the bulk phase increased, for 10% DOPA/90% MO membrane in excess water, a Q(229) to Q(224) phase transition occurred at 60 mM NaCl, and then a Q(224) to H(II) phase transition occurred at 1.2 M NaCl. Similarly, for 30% DOPA/70% MO membrane in excess water, at low NaCl concentrations it was in the L(alpha) phase, but at and above 0.50 M NaCl it was in the Q(224) phase, and then at 0.65 M NaCl a Q(224) to H(II) phase transition occurred. These results indicate that the electrostatic interactions in the membrane interface make the Q(229) phase more stable than the Q(224) phase, and that, at larger electrostatic interactions, the L(alpha) phase is more stable than the cubic phases (Q(224) and Q(229)). We have found that the addition of tetradecane to the MO membrane induced a Q(224)-to-H(II) phase transition and also that to the 30% DOPA/70% MO membrane induced an L(alpha)-to-H(II) phase transition. By using these membranes, the effect of the electrostatic interactions resulting from the membrane surface charge (DOPA) on the spontaneous curvature of the monolayer membrane has been investigated. The increase in DOPA concentration in the DOPA/MO membrane reduced the absolute value of spontaneous curvature of the membrane. In the 30% DOPA/70% MO membrane, the absolute value of spontaneous curvature of the membrane increased with an increase in NaCl concentration. On the basis of these new results, the phase stability of DOPA/MO membranes can be reasonably explained by the spontaneous curvature of the monolayer membrane and a curvature elastic energy of the membrane.

Glycerides↗