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Cell cycle time, growth fraction and cell loss in xenografted head and neck cancer.

In a previous cell kinetic study with DNA flow cytometry (FCM) on xenografted human squamous cell carcinoma of the head and neck (HNSCC), there were significant variations in cell cycle phase distribution during early growth of the transplanted tumors. The object of the present study was to investigate further the cell kinetic mechanisms associated with variation in the growth rate of HNSCC. Xenografts from the same tumor line were examined on three different occasions: 12, 25 and 36 days following transplantation into nude mice. Cell cycle time "Tc", tumor growth fraction (GF) and cell loss factor were determined by computerized curve-fit analysis from percentage labelled mitoses curves. The mean growth curve of the tumors was logistic, with tumor doubling time (Td) ranging from 7.39 days to 8.86 days, and increasing as tumor volume increased. Although growth rate was higher in younger tumors, the cell cycle was longer with a median "Tc" of 69 hours on day 12 versus 34 hours on day 36. There were only minor variations in cell cycle phase distribution as measured by FCM. The growth fraction decreased slightly from 80% to 69% between day 12 and day 36, whereas cell loss factor increased markedly from 49% on day 12 to 78% on day 36. The results of the study emphasize the fact that cell loss is one of the most important cell kinetic determinants of tumor growth rate. This should be borne in mind when calculating proliferative cell kinetics in HNSCC.

Animals↗

Effect of masker level on overshoot in running- and frozen-noise maskers.

Masked thresholds were measured with running- and frozen-noise maskers. The 5-kHz signal was 2 ms in duration. The masker was low-pass noise (20 Hz-10 kHz); its total duration was 300 ms. The overall level of the masker was 30, 50, or 70 dB SPL. The onset of the signal was delayed by 0, 3, 8, 18, 198, or 278 ms relative to the onset of the masker. In all frozen-noise measurements, the signal was added to the same fine structure of the noise. Overshoot in frozen noise was measured for two starting phases of the signal that led to a 10-dB difference for large signal-onset delays. In all three configurations (running noise and frozen noise with two different signal phases) masker level had a similar influence on overshoot. At the intermediate masker level (50 dB SPL), a significant amount of overshoot (up to 15 dB) was observed in all three conditions. At the low and the high masker levels, overshoot was very much reduced, and even became negative in most conditions for the 30-dB-SPL masker. For the 50-dB frozen-noise masker, the total variation of thresholds with signal phase was 8 to 11 dB for long signal-onset delays, but only 3 to 6 dB for short delays. For the low- and high-level maskers, where only a small overshoot was observed, the threshold variation with phase for a signal at masker onset was the same as that for the long-delay condition. An explanation for the variation of signal detectability with masker level is proposed that refers explicitly to the compressive input-output characteristic of the basilar membrane at intermediate levels.

Adult↗

Stabilization and relative phase effects in a dichromatically driven diatomic Morse molecule: interpretation based on nonlinear classical dynamics.

The dissociation dynamics of a dichromatically laser-driven diatomic Morse molecule vibrating in the ground state is investigated by applying tools of the nonlinear theory of classical Hamiltonian systems. Emphasis is placed on the role of the relative phase of the two fields, phi. First, it is found that, just like in quantum mechanics, there is dependence of the dissociation probability on phi. Then, it is demonstrated that addition of the second laser leads to suppression of probability (stabilization), when the intensity of the first laser is kept constant just above or below the single laser dissociation threshold. This "chemical bond hardening" diminishes as phi increases. These effects are investigated and interpreted in terms of modifications in phase space topology. Variations of phi as well as of the intensity of the second laser may cause (i) appearance/disappearance of the stability island corresponding to the common resonance with the lowest energy and (ii) deformation and movement of the region of Kolmogorov-Arnold-Moser tori that survive from the undriven system. The latter is the main origin in phase space of stabilization and phi dependence. Finally, it is shown that the use of short laser pulses enhances both effects.

Journal Article↗

Interaction of two variable proteins (PilE and PilC) required for pilus-mediated adherence of Neisseria gonorrhoeae to human epithelial cells.

Pili confer the initial ability of Neisseria gonorrhoeae to bind to epithelial cells. Pilin (PilE), the major pilus subunit, and a minor protein termed PilC, reportedly essential for pilus biogenesis, undergo intra-strain phase and structural variation. We demonstrate here that at least two different adherence properties are associated with the gonococcal pili: one is specific for erythrocytes, which is virtually unaffected by PilE variation, and another is specific for epithelial cells, and is modulated in response to the variation of PilE. Based on this finding, mutants of a recA- strain were selected that had lost the ability to bind to human cornea epithelial cells (A-) but retained the ability to form pili (P+) and to agglutinate human erythrocytes (H+). The adherence-negative mutants failed to produce detectable levels of PilC1 or PilC2 proteins, representing piIC phase variants generated in the absence of RecA. The A- pilC phase variants were indistinguishable from their A+ parents and spontaneous A+ revertants with regard to the amount of PilE produced and its electrophoretic mobility, the degrees of piliation and haemagglutination, and the pilE nucleotide sequence. These data demonstrate a central role for PilC in pilus-mediated adherence of N. gonorrhoeae to human epithelial cells and further indicate that neither PilC1 nor PilC2 is obligatory for the assembly of gonococcal pili.

Amino Acid Sequence↗

Blood flow imaging through detection of temporal variations in magnetization.

Two hypotheses were tested: (a) that view-to-view variations in bulk phase and modulus of magnetization in vascular volume elements can indicate the presence of disordered blood flow, and (b) that a substantial loss of signal intensity on magnetic resonance (MR) angiograms of poststenotic regions is due to view-to-view changes in magnetization. To test these hypotheses, a technique was developed in which view-to-view variations in transverse magnetization were used to create angiographic projection images, which showed only disordered flow (disordered flow maps) in vitro and in vivo. In phantom studies, this technique recovered signal intensity downstream from stenoses. A combination of disordered flow maps with morphologic images improved visualization of stenotic regions and provided information on characteristics of local flow. These results show that view-to-view variations in transverse magnetization occur in regions of disordered flow and are an important cause of loss of signal intensity. This technique can provide information about dynamic blood flow and improve depiction of anatomic structures on MR angiograms.

Blood Circulation↗

Pleomorphic adenoma of the salivary glands: prospective clinicopathologic and flow cytometric study.

BACKGROUND: Primary surgery is the appropriate treatment for most pleomorphic adenomas of the salivary glands. However, no reliable criteria have been established to predict tumors with aggressive features. METHODS: In 44 patients treated for salivary pleomorphic adenoma, clinicopathologic features and flow cytometric findings were recorded. S-phase fractions were classified as low (0%-1%), intermediate (1.1%-5%), and high (>5%). The relationship between clinicopathologic parameters and flow cytometric DNA analysis was assessed with the Pearson's correlation coefficient. RESULTS: The mean tumor size was 24.4 mm. Microscopically, myxoid structures predominated in 61.4% of cases. Only five specimens showed high S-phase fractions. All samples were DNA diploid (mean coefficient of variation 3.2%). S-phase fraction and tumor size correlated significantly (r = 0.73, P = 0.003). Two patients with recurrent tumors had high S-phase fractions both on the first resected specimens and at the time of the second operation. CONCLUSIONS: The S-phase fraction may be a valuable predictor of aggressive behavior in salivary pleomorphic adenomas. High S-phase fractions were associated with higher tumor size and a greater tendency to recur.

Adenoma, Pleomorphic↗

Direct high-performance liquid chromatographic separation of unusual secondary amino acids and a comparison of the performances of Chirobiotic T and TAG columns.

Two macrocyclic glycopeptide antibiotic-type chiral stationary phases (CSPs) based on native teicoplanin and teicoplanin aglycone, Chirobiotic T and TAG, respectively, were evaluated with regard to the high-performance liquid chromatographic separation of the enantiomers of 10 secondary alpha-amino acids (imino acids). The chromatographic results are given as the retention, separation and resolution factors, together with the enantioselective free energy difference corresponding to the separation of the enantiomers. By application of these two CSPs, excellent resolutions were achieved for the investigated compounds by using reversed-phase mobile mode systems. The separation conditions were optimized by variation of the mobile phase composition. The difference in enantioselective free energy between the aglycone CSP and the teicoplanin CSP for these particular amino acids ranged between 0.70 and -1.83 kJ mol(-1). It was established that better enantioseparations of the secondary alpha-amino acids were attained in most cases on the aglycone CSP.

Amino Acids↗

Elongated versions of Vlp surface lipoproteins protect Mycoplasma hyorhinis escape variants from growth-inhibiting host antibodies.

Variation in Vlp surface proteins of Mycoplasma hyorhinis was evaluated in terms of its role in determining susceptibility of organisms to growth inhibition by host antibodies (Abs). High-frequency switching of Vlp surface lipoproteins has been studied in isogenic lineages of M. hyorhinis SK76. In these lineages, the products of three genes, vlpA, vlpB, and vlpC, are subject to phase and size variation in vitro, which occur through distinct mutator elements that independently govern the expression of each vlp gene (promoter mutations) or the size of the vlp gene product (by intragenic expansion or contraction of a 3' region containing tandem repeats). Isogenic clonal variants of M. hyorhinis SK76 expressing distinct profiles of Vlp products were assessed for their susceptibility to complement-independent growth inhibition by serum Abs of swine experimentally infected with the arthritigenic SK76 strain. Invariably, variants expressing longer versions of VlpA, VlpB, or VlpC (each expressed individually) were completely resistant to host immune serum Abs, whereas variants expressing shorter allelic versions of each Vlp were susceptible. The target of growth-inhibiting Abs was not the Vlp products, since removal of anti-Vlp Abs had no effect on the inhibitory activity of the host immune serum on susceptible variants. Escape variant populations derived by propagating susceptible variants in an immune (versus control) host serum revealed a strong selection for the long-Vlp phenotype, irrespective of the identity of the Vlp expressed. Apparent mutational pathways of acquiring the protective phenotype included mutational switches to express long vlp genes that had been transcriptionally silent or switches to elongate expressed vlp genes. These results suggest that a major function of the Vlp system is to shield the wall-less mycoplasma surface from host Abs capable of binding vital (and as-yet-unidentified) surface antigens of this organism.

Alleles↗

["Late" obstruction induced by physical exertion in 15 boys with asthma caused by food aeroallergens in 2 periods of the year (winter and summer)].

The aim of this study was the observation of the presence of the "late" phase of obstruction after free run in subjects responsive either to aeoallergens either to food allergens but with withdrawal of food allergens from diet in comparison with control group of children which did not have the food allergens exclusion. The study was executed in February and in June in the same children's group having the purpose to observe the variation of this phase in June with a great pollination of graminacee. The results showed the presence of a late phase of obstruction of magnitude similar to the quick phase which begun from III to VII h. but was no greater than -20% FEV1 fall. The control group showed in many children the fall of -20% FEV1 fall. The group of children responsive to the graminacee with food allergen's exclusion showed in June a greater obstructive late phase in comparison to February but the obstruction never was greater than -20% FEV1 fall.

Adolescent↗

DNA flow cytometry of fresh and paraffin-embedded tissue using cytokeratin staining.

DNA flow cytometry measurements were performed using cytokeratin as a second parameter to identify epithelial cells selectively in fresh and in archival paraffin samples of normal and adenocarcinoma tissues from breast and colon. Fresh specimens consisted of 22 adenocarcinomas of breast, 20 adenocarcinomas of colon, 16 control breast samples, and 13 control colon samples. Paraffin block specimens consisted of 22 adenocarcinomas of breast (the same as fresh samples), 20 adenocarcinomas of colon (the same as fresh samples), 37 control breast samples and 34 control colon samples. The average proportion of cytokeratin-positive cells per group ranged from 31 to 55% for fresh samples and from 14 to 34% for paraffin samples. For aneuploid cells populations of adenocarcinomas, which consist only of epithelial cells, the average percentage of cytokeratin-positive cells ranged from 60 to 72%. The technique gave satisfactory measurements of ploidy and of cell cycle data in both types of samples. Cell cycle measurements were less accurate than ploidy measurements in both types of samples, and multiple sampling will be required for adequate accuracy. The average S-phase fraction of cytokeratin-positive cells ranged from 6 to 15% for fresh specimens and from 11 to 20% for paraffin samples. Similar data were obtained for the proliferative index (G1 + S + G2 + M phases). The coefficients of variation were smaller for proliferative index than for S-phase fraction data, indicating greater accuracy. Paraffin data give higher cycling cell measurements than corresponding fresh data, so separate standardization of measurements may be required for fresh and for paraffin data.

Adenocarcinoma↗

Experimental cardiac tamponade: a hemodynamic and Doppler echocardiographic reexamination of the relation of right and left heart ejection dynamics to the phase of respiration.

A hallmark of cardiac tamponade is pulsus paradoxus. However, the exact mechanism of pulsus paradoxus and the relation of left and right ventricular ejection dynamics remain controversial, with some studies suggesting an inverse relation in ventricular filling and ejection and others citing a more important role for the effects of right heart ejection dynamics delayed by transit through the pulmonary artery bed. To specifically reexamine this issue, six sedated but spontaneously breathing dogs were studied during experimental cardiac tamponade with use of extensive hemodynamic instrumentation and Doppler methods. During cardiac tamponade, left ventricular systolic pressure decreased from 125.8 +/- 12.1 to 81.7 +/- 26.7 mm Hg (p less than 0.01) and cardiac output from 5.86 +/- 1.48 to 2.34 +/- 0.98 liters/min (p less than 0.001); mean pericardial pressure increased from -1.2 +/- 0.8 to 10.5 +/- 3 mm Hg (p less than 0.001) and pulsus paradoxus from 4.3 +/- 1.6 to 10.7 +/- 1.2 mm Hg (p less than 0.001) compared with baseline values. An inverse relation in left and right ventricular ejection dynamics that was very close to 180 degrees out of phase was seen throughout the respiratory cycle in multiple hemodynamic and Doppler variables including peak systolic pressures, aortic and pulmonary flow velocities and ventricular ejection times. Simultaneous recording of the transmitral pressure gradient provided indirect evidence that the ventricular ejection dynamics were directly related to changes in ventricular filling. However, the magnitude of ventricular pressure or output flow velocity for each respiratory cycle was variable, depending on the exact timing of filling and ejection in relation to the phase of respiration. Variation in left ventricular output due to changes in right ventricular output delayed by transit through the pulmonary vasculature was not recognized in any animal. It is concluded that in spontaneously breathing dogs with acute cardiac tamponade, peak ventricular pressures, ventricular ejection times and pulmonary and aortic flow velocities have an inverse relation that is very close to 180 degrees out of phase.

Animals↗

Coherent phase control of the product branching ratio in the photodissociation of dimethylsulfide.

Coherent phase control of the photodissociation reaction of the dimethylsulfide has been achieved by means of quantum-mechanical interference between one- and three-photon transitions. Dimethylsulfide was irradiated by fundamental and frequency-tripled outputs of a visible laser (600.5-602.5 nm), simultaneously to yield CH3S+ and CH3SCH2+ fragment ions. The branching ratio of the two product channels could be modulated with variation of the phase difference between the light fields. This accounted for the difference between the molecular phases of the two product channels. The phase lag was observed to have a maximum value of 8 degrees at 601.5 nm. This is the first result of a selective bond breaking in a polyatomic molecule by the coherent phase control.

Journal Article↗

Effects of harmonic phase on nonlinear surface acoustic waves in the (111) plane of cubic crystals.

Spectral evolution equations are used to perform numerical studies of nonlinear surface acoustic waves in the (111) plane of several nonpiezoelectric cubic crystals. Nonlinearity matrix elements which describe the coupling of harmonic interactions are used to characterize velocity waveform distortion. In contrast to isotropic solids and the (001) plane of cubic crystals, the nonlinearity matrix elements usually cannot be written in a real-valued form. As a result, the harmonic components are not necessarily in phase, and dramatic variations in waveforms and propagation curves can be observed. Simulations are performed for initially monofrequency surface waves. In some directions the waveforms distort in a manner similar to nonlinear Rayleigh waves, while in other directions the velocity waveforms distort asymmetrically and the formation of shocks and cusped peaks is less distinct. In some cases, oscillations occur near the shocks and peaks because of phase differences between harmonics. A mathematical transformation based on the phase of the matrix elements is shown to provide a reasonable approximation of asymmetric waveform distortion in cases where the matrix elements have similar phase.

Journal Article↗

Intrastrain and interstrain genetic variation within a paralogous gene family in Chlamydia pneumoniae.

BACKGROUND: Chlamydia pneumoniae causes human respiratory diseases and has recently been associated with atherosclerosis. Analysis of the three recently published C. pneumoniae genomes has led to the identification of a new gene family (the Cpn 1054 family) that consists of 11 predicted genes and gene fragments. Each member encodes a polypeptide with a hydrophobic domain characteristic of proteins localized to the inclusion membrane. RESULTS: Comparative analysis of this gene family within the published genome sequences provided evidence that multiple levels of genetic variation are evident within this single collection of paralogous genes. Frameshift mutations are found that result in both truncated gene products and pseudogenes that vary among isolates. Several genes in this family contain polycytosine (polyC) tracts either upstream or within the terminal 5' end of the predicted coding sequence. The length of the polyC stretch varies between paralogous genes and within single genes in the three genomes. Sequence analysis of genomic DNA from a collection of 12 C. pneumoniae clinical isolates was used to determine the extent of the variation in the Cpn 1054 gene family. CONCLUSIONS: These studies demonstrate that sequence variability is present both among strains and within strains at several of the loci. In particular, changes in the length of the polyC tract associated with the different Cpn 1054 gene family members are common within each tested C. pneumoniae isolate. The variability identified within this newly described gene family may modulate either phase or antigenic variation and subsequent physiologic diversity within a C. pneumoniae population.

5' Flanking Region↗

[Features of volume and pressure pulsations during changes in "reserve space" within the cranial cavity].

Changes in the amplitude and phase characteristics of pulse variations of volumes and pressures in response to an increase in the intracranial pressure are discussed. The study of 60 neurosurgical patients has shown that as the intracranial (subdural) pressure grows the amplitude of pulse variations of subdural pressure increase to 30 mm Hg and then decreases, the phase characteristics of the pulse waves changing accordingly. The changes in the amplitudes of pulse variations produced by mechanical displacement of the walls of cerebral lateral ventricles measured with the aid of the noninvasive ultrasonic method suggest that an increase in the CSF pressure above 200 mm H2O makes the intracranial cavity too "tight", which depletes the reserve spaces of the cerebrospinal system. If the above changes develop consistently, then it can be assumed that each of the pressure ranges isolated can be characterized by amplitude-phase parameters of pulse waves that form a specific pattern. Taking into consideration the potentials of noninvasive measurements of amplitude-phase characteristics of the intracranial pulse wave by means of ultrasonic echolocation, it deems possible to develop noninvasive techniques for measuring compensatory abilities of the cerebrospinal system.

Biomechanical Phenomena↗

A limit cycle mathematical model of the REM sleep oscillator system.

A limit cycle mathematical model of the rapid-eye-movement (REM) sleep oscillator system has been developed from a structural model of interaction of populations of REM-on and REM-off neurons. The marked differences in latency, amplitude, and duration of the first REM sleep period seen with circadian variation and depressive pathology are modeled by beginning the REM oscillation at different initial points relative to the final position in the limit cycle. Beginning from a point that is graphically interior to the limit cycle produces a long-latency, short-duration, and less intense first REM period. Beginning from a point graphically exterior to the limit cycle produces a short-latency, long-duration, and more intense first REM period. In the model the determinant of whether the oscillation begins exterior or interior to the limit cycle is the time course of decay of the REM-off population discharge activity at sleep onset. When this time course is made to depend on circadian phase, the model produces a very close match to the empirically observed large shifts between the first and second REM periods in duration (often a 50% change) and intensity and also closely mimics the empirically observed shifts in REM latency as human sleep begins at different circadian phases. Although this variation in limit cycle entry accounts for the major changes in REM sleep over the night, the model also postulates a continuous but small circadian variation (of the order of +/- 5% change in REM parameters) acting throughout the course of a night's sleep. Because the model is derived from actual physiological data, rather than being a purely ad hoc or phenomenological construct, it offers the possibility of direct tests of its postulates through neurobiological studies in animals, by circadian phase-related manipulations of the sleep cycle, and through perturbations of the system in humans by the use of drugs. Indeed, an explicit phase-response curve of the system to cholinergic agonists has been developed; this will permit experimental tests of the model in both animals and humans.

Animals↗

Is coronary sinus blood oxygen tension behavior determined by myocardial oxygen tension variation during cardiac reperfusion?

The relationship between coronary sinus blood oxygen tension (CSPO2) and myocardial oxygen tension (MPO2) variations during cardiac ischemia and reperfusion was studied in anesthetized open-chest dogs. Oxygen tension was measured by a polarographic method. Ischemia resulted in a slightly decreased CSPO2 and a more pronounced reduction of MPO2. After reperfusion the CSPO2 rose rapidly and transiently before it returned gradually to the control level. By contrast, during the recovery period, the MPO2 increased slowly, with recovery occurring long after the peak of CSPO2. These data suggest that during the reperfusion phase, the CSPO2 variation is probably due to opening of the myocardial arteriovenous shunts instead of an increase of flow through the myocardial capillary bed.

Animals↗

Nycthemeral patterns of thyroid hormones and their relationships with thyrotropin variations and sleep structure.

In order to precise the relationships between TSH, FT3, and FT4 nycthemeral variations and the relationships between thyroid hormone variations and sleep, 8 healthy young males were studied twice, once during a 24-h experiment with normal nocturnal sleep, and once during a night of sleep deprivation. The subjects received continuous enteral nutrition and remained supine during the whole experiment. Blood was sampled every 10 min for TSH, FT3, and FT4 measurements. Thyroid hormones exhibited small oscillations which were not systematically related to TSH pulses, and there was no evidence of a nycthemeral rhythm. SWS was associated with TSH declining phases, whereas awakenings were strongly associated with ascending phases of TSH variations. There was no association between sleep structure or awakenings and thyroid hormones. Sleep deprivation led to increased TSH and FT3 levels, without any variation in FT4 levels. These results demonstrate that short-term thyroid hormone variations do not only depend on the effect of TSH on thyroid secretion but also on a possible role of TSH on peripheral FT4 to FT3 conversion. Conversely, the relationships between TSH and SWS or awakenings are not mediated by thyroid hormones.

Adult↗