DNA electrodiffusion in a 2D array of posts.
Explore the source record for details and available documents.
Journal Article↗
Biomedical subjects
A Szabo
Publications and source records attributed to A Szabo.
Erratum: Time-domain violation of the optical Bloch equations for solids
Explore the source record for details and available documents.
Journal Article↗
Mental challenge elicits "additional" increases in heart rate during low and moderate intensity cycling.
The combined effects of exercise and mental challenge on heart rate (HR) and T-wave amplitude (TWA) were studied. Twenty male students cycled at low (40% of their relative maximal HR reserve (MHRR), for 15 min) and medium (60% MHRR, for 10 min) intensity exercise workloads. Subjects performed a series of mental arithmetic problems for one minute each time: two min before cycling, 10 min into low intensity cycling, 10 min into medium intensity cycling, and two and 20 min, respectively, after cycling. During both exercise workloads, the mental arithmetic elicited significant additional increases in HR. Although TWA decreased in response to mental arithmetic at rest, no changes in TWA were observed in response to mental task during exercise. However, TWA increased significantly following the mental challenge. These results suggest that even a mild mental challenge is capable of inducing further changes in the autonomic response during low and moderate exercise.
Adult↗
Simulation of the kinetics of ligand binding to a protein by molecular dynamics: geminate rebinding of nitric oxide to myoglobin.
We have begun to use molecular dynamics to simulate the kinetics of nitric oxide rebinding to myoglobin after photodissociation. Rebinding was simulated using a potential function that switches smoothly between a nonbinding potential and a binding potential as a function of the position and orientation of the ligand, with no barrier arising from the crossing of potential surfaces of different electron spin. In 96 of 100 trajectories, the ligand rebound in < 15 ps. The kinetic progress curve was obtained by determining the time in each trajectory at which the ligand rebound and then calculating the fraction of unbound ligands as a function of time. The curve can be well reproduced by a simple model based on the dynamics of a Langevin particle moving on a one-dimensional potential of mean force calculated from nonreactive protein trajectories. The rate of escape from the energy well adjacent to the heme is in good agreement with the value calculated from experimental data, suggesting that a multiple-well model provides a plausible explanation for the nonexponential rebinding kinetics. A transition-state analysis suggests that protein conformational relaxation coupled to the displacement of the iron from the heme plane is an unlikely cause for the nonexponential rebinding of nitric oxide.
Computer Simulation↗
Time-domain violation of the optical Bloch equations for solids.
Explore the source record for details and available documents.
Journal Article↗
Antigen presentation capacity of murine macrophages infected with Leishmania amazonensis amastigotes.
Leishmania-infected M phi are potential candidates for the presentation of parasite Ag to Leishmania-specific CD4+ T lymphocytes. To assess whether infected cells could function as APC, we examined the ability of bone marrow-derived M phi infected with Leishmania amazonensis amastigotes to stimulate various CD4+, l-Ad- or l-Ed-restricted T-cell hybridomas specific for the bacteriophage lambda repressor cl protein, the human chorionic gonadotropin or OVA. A reduced capacity of infected M phi to present native Ag to most T-cell hybridomas tested was noted that was probably a result of a lower expression on their plasma membrane of stimulatory [la-peptide] complexes. Neither a reduced Ag uptake nor an altered Ag processing appeared to be at the origin of the partial inability of infected M phi to present Ag. As regards the level of plasma membrane la expression, no quantitative difference could be detected between uninfected and infected M phi. Moreover, after fixation with paraformaldehyde, the ability of plasma membrane la molecules to bind immunogenic peptides was apparently not reduced in infected M phi. So, these cells most likely expressed functional la molecules on their cell surface. Interestingly, infected M phi and M phi infected then cured by a treatment with a leishmanicidal compound were similarly impaired in their capacity to present native Ag or peptides to the hybridomas, and no recovery was noted even 24 h after the leishmanicidal treatment. Furthermore, infected M phi and M phi incubated with heat-killed amastigotes or with an amastigote homogenate exhibited similar inhibitions of Ag presentation. Taken together, these results suggest that the functional failure of infected M phi to present exogenous Ag could be because either of interferences with the events leading to the meeting of la molecules with peptides derived from these exogenous Ag or to a competition for binding to la molecules between these peptides and parasite molecules.
Animals↗
1,25-dihydroxyvitamin D3 production and vitamin D3 receptor expression are developmentally regulated during differentiation of human monocytes into macrophages.
It has been well established that human mononuclear phagocytes have the capacity to produce 1,25-dihydroxy-vitamin D3 [1,25(OH)3D3] and express the vitamin D receptor (VDR). However, 1 alpha-hydroxylase activity and VDR receptor expression during differentiation of monocytes (MO) into mature macrophages (MAC) have not been previously examined. The in vitro maturation of blood MO can serve as a model for the in vivo transformation of immature blood MO into MAC. Here, when cultured in the presence of serum, MO undergo characteristic changes in morphology, antigenic phenotype, and functional activity consistent with their differentiation into MAC. We serially measured 1,25(OH)2D3 and 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] synthesis, specific [3H]-1,25(OH)2D3 binding, and VDR mRNA levels during in vitro maturation of MO into MAC and correlated these functions with maturation-associated changes in the phenotype (MAX.1 and CD71) and secretory repertoire (interleukin-1 beta [IL-1 beta], neopterin) of the cells. MO showed only little conversion of 25-(OH)D3 into 1,25(OH)2D3 (1.4 +/- 0.4 pmol/10(6) cells/6 h, n = 5) that increased gradually during maturation into MAC at day 8 of culture (5.3 +/- 4.3 pmol/10(6) cells/6 h, n = 5). Interferon-gamma (IFN-gamma) increased baseline 1,25(OH)2D3-synthesis approximately twofold during all phases of differentiation. The time course of increased 1,25(OH)2D3-synthesis correlated with enhanced secretion of neopterin and expression of MAX.1 and CD71. The addition of exogenous 1,25(OH)2D3 did not influence constitutive 1,25(OH)2D3 synthesis, but IFN-gamma-stimulated production was suppressed to baseline levels. Exogenous 1,25(OH)2D3 also stimulated 24,25(OH)2D3 synthesis in freshly isolated MO (from 1.0 +/- 0.8 pmol/6 h to 5.6 +/- 0.9 pmol), whereas matured MAC showed no 24,25(OH)2D3 synthesis. Furthermore, we examined the expression of the VDR during the differentiation process. VDR mRNA and protein were constitutively expressed in MO, whereas VDR was downregulated in mature MAC on both the mRNA and protein levels. Homologous upregulation of VDR protein by 1,25(OH)2D3 occurred in MO and, to a lesser degree, in MAC. In contrast, VDR mRNA concentrations were not influenced by 1,25(OH)2D3. Taken together, our results show that MO into MAC differentiation in vitro is associated with (1) an enhanced capacity to synthesize 1,25(OH)2D3, (2) a loss of 24,25(OH)2D3-synthesizing activity, and (3) a decrease in the expression of VDR mRNA and protein. Because 1,25(OH)2D3 was shown to induce differentiation of MO into MAC, our data sugest an autoregulatory mechanism of MO/MAC generation by 1,25(OH)2D3.
24,25-Dihydroxyvitamin D 3↗
The combined effects of orthostatic and mental stress on heart rate, T-wave amplitude, and pulse transit time.
The purpose of the present study was to determine whether cardiovascular reactivity to mental stress may be comparable in sitting and standing postures. Fifteen healthy males performed two 1-min mental arithmetic tasks, either while sitting or while standing, in counterbalanced order. Heart rate, pulse transit time, and T-wave amplitude were recorded in the last 10 s of the minute before, during, and after the mental arithmetic. Reactivity scores for each of the dependent measures were computed by calculating the percentage change from baseline values. Data were analyzed with multivariate and univariate repeated measures analysis of variance. Heart rate reactivity to the combination of orthostatic and mental stress was greater than to either stressor alone. Cardiac-sympathetic reactivity was greater in response to orthostatic than to mental stress as revealed by greater changes in T-wave amplitude and pulse transit time in response to the former. No additional decreases in T-wave amplitude, in response to mental stress, were observed during standing, but pulse transit time decreased in the same situation. However, no changes in pulse transit time in response to mental stress were observed in the sitting position. These results demonstrate that cardiovascular reactivity to mental stress depends on the body position in which the stressor is encountered.
Adult↗
Psychophysiological profiles in response to various challenges during recovery from acute aerobic exercise.
Cardiovascular and sympathetic profiles in response to a series of physical and mental challenges were examined during recovery from an acute bout of aerobic exercise performed at 60% VO2,max for 30 min. 9 healthy men were tested on two occasions, once under an experimental (exercise) and once under a control (video watching) condition, in a counter-balanced order, one week apart. Although no differences in blood pressure were found in the two conditions, heart rate and plasma catecholamine concentrations were higher in the exercise than in the control session. These findings were partly attributed to elevated physiological levels 'carried over' from exercise. State anxiety and anger-hostility, however, were lower in the post-experimental period than in the pre-experimental period. The results are discussed in terms of their relevance to exercise and stress psychophysiology.
Adult↗
Fluorescence dequenching kinetics of single cell-cell fusion complexes.
In an earlier paper which models the cell-cell (or virus-cell) fusion complex as two partial spherical vesicles joined at a narrow neck (Rubin, R. J., and Yi-der Chen. 1990. Biophys. J. 58:1157-1167), the redistribution by diffusion of lipid-like molecules through the neck between the two fused cell surfaces was studied. In this paper, we extend the study to the calculation of the kinetics of fluorescence increase in a single fusion complex when the lipid-like molecules are fluorescent and self-quenching. The formalism developed in this paper is useful in deducing fusion activation mechanisms from cuvette fluorescence measurements in cell-cell fusion systems. Two different procedures are presented: 1) an exact one which is based on the exact local density functions obtained from diffusion equations in our earlier study; and 2) an approximate one which is based on treating the kinetics of transfer of probes between the two fused cells as a two-state chemical reaction. For typical cell-cell fusion complexes, the fluorescence dequencing curves calculated from the exact and approximate procedures are very similar. Due to its simplicity, the approximate method should be very useful in future applications. The formalism is applied to a typical cell-cell fusion complex to study the sensitivity of dequenching curves to changes in various fusion parameters, such as the radii of the cells, the radius of the pore at the fusion junction, and the number of probes initially loaded to the complex.
Animals↗
Theory of photoselection by intense light pulses. Influence of reorientational dynamics and chemical kinetics on absorbance measurements.
The theory of absorbance measurements on a system (e.g., chromophore(s) in a protein) that undergoes a sequence of reactions initiated by a linearly polarized light pulse is developed for excitation pulses of arbitrary intensity. This formalism is based on a set of master equations describing the time evolution of the orientational distribution function of the various species resulting from excitation, reorientational dynamics, and chemical kinetics. For intense but short excitation pulses, the changes in absorbance (for arbitrary polarization directions of the excitation and probe pulses) and the absorption anisotropy are expressed in terms of reorientational correlation functions. The influence of the internal motions of the chromophore as well as the overall motions of the molecules is considered. When the duration of the excitation pulse is long compared to the time-scale of internal motions but comparable to the overall correlation time of the molecule that is reorienting isotropically, the problem of calculating the changes in absorbance is reduced to the solution of a set of first-order coupled differential equations. Emphasis is placed on obtaining explicit results for quantities that are measured in photolysis and fluorescence experiments so as to facilitate the analysis of experimental data.
Binding Sites↗
Intermittent versus continuous administration of 1,25-dihydroxyvitamin D3 in experimental renal hyperparathyroidism.
Conflicting results have been reported regarding the efficacy of intermittent versus continuous administration of 1,25(OH)2D3 in renal secondary hyperparathyroidism. To address this issue we examined sham-operated control rats and hyperparathyroid rats with subtotal (5/6) nephrectomy (Nx). The Nx animals (20 to 22 animals per group) were subjected to three treatment protocols: (i) solvent treatment (Nx-solvent); (ii) two i.p. injections of 35 pmol 1,25(OH)2D3 on days 0 and 4 (Nx-bolus); and (iii) continuous infusion of 70 pmol 1,25(OH)2D3 over six days via osmotic minipump (Nx-infusion). All measurements were performed six days after start of treatment. As compared to sham-operated controls, the pre-pro-PTH/beta-actin mRNA ratio was 2.04-fold higher in Nx-solvent. Both modes of administration of 1,25(OH)2D3 resulted in inhibition of PTH mRNA concentrations relative to Nx-solvent. The pre-pro-PTH/beta-actin mRNA ratio was, however, significantly lower (P < 0.05) in Nx-bolus than in Nx-infusion (Nx-bolus 1.26 higher than sham-operated controls; Nx-infusion 1.65 higher than sham-operated controls). Aminoterminal PTH (N-PTH) serum concentrations were higher in Nx-solvent (52 +/- 4 pg/ml) than in sham-operated controls (32 +/- 3 pg/ml, P < 0.01). N-PTH concentrations in Nx-bolus (38 +/- 4 pg/ml) were significantly lower than in Nx-solvent (P < 0.01) and in Nx-infusion (46 +/- 4 pg/ml, P < 0.05). Parathyroid gland weight (microgram/g body wt) was higher in Nx-solvent (1.30 +/- 0.08 pg/ml) than in sham-operated controls (0.79 +/- 0.04 pg/ml, P < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
A safe simple method for removal of the gallbladder through the umbilical trocar site during laparoscopic cholecystectomy.
After laparoscopic resection, the gallbladder must be removed, usually through the umbilical trocar site. Occasionally, there is difficulty in removing the gallbladder from the abdominal cavity due to its large dimensions or its contents. There are two main options to extract the gallbladder in such cases. One is evacuation of its content through the gallbladder neck protruding from the abdominal wall. The second is extension of the periumbilical fascial opening. In the first method, gallbladder content might sometimes spill into the peritoneal cavity or contaminate the incision site. In the second, there might not be enough control concerning the extent of injury caused to the periumbilical abdominal wall during enlargement of the fascial opening. A simple precise and safe method is suggested to enlarge the fascial incision by direct vision using transillumination.
Abdominal Muscles↗
Parathyroidectomy in secondary (renal) hyperparathyroidism--whom, when, how?
Explore the source record for details and available documents.
Calcitriol↗
A 500 ps molecular dynamics simulation study of interleukin-1 beta in water. Correlation with nuclear magnetic resonance spectroscopy and crystallography.
We report the results of a 500 ps molecular dynamics simulation of the cytokine interleukin-1 beta, a protein of 153 amino acids, immersed in a sphere of 3783 bulk water molecules with a radius of 33 A. The simulation reproduces the amplitudes of the fast librational motions of the backbone N-H bonds determined from 15N nuclear magnetic relaxation data, as well as the crystallographic B-factors. Moreover, this study suggests a molecular picture of the nature of the slow internal motions that have been inferred from nuclear magnetic resonance relaxation experiments. These experiments indicated that, in addition to fast motions common to all residues, 32 surface residues exhibit slow motions on the 400 ps to 5 ns time-scale. While the present simulation is not sufficiently long to provide a quantitative description of events on this time-scale, it is long enough to observe several large amplitude transitions that are likely candidates for these slow motions. Specifically, in many of these 32 residues, the N-H groups are hydrogen bonded and infrequent dihedral transitions cause the N-H vectors to jump between states with well-defined orientations. It is shown that the time course of the angular reorientational correlation functions of these residues calculated from the trajectory is a reflection of the random times at which these infrequent jumps happen to have occurred. Thus, while the rate of these transitions cannot be quantified, the simulated decay of these correlation functions is completely consistent with the physical picture in which the N-H vectors, in addition to fast librational motion, undergo large amplitude jumps between conformations stabilized by hydrogen bonds.
Computer Simulation↗
Sarcoplasmic reticulum from rabbit and winter flounder: temperature-dependence of protein conformation and lipid motion.
A comparative study of lipids and proteins in sarcoplasmic reticulum (SR) from rabbit and flounder has been undertaken. The protein/phospholipid ratio (w/w) was 3:1 in flounder SR (FSR) and 2.2:1 in rabbit SR (RSR). Both membranes had similar contents of PC (70%) and PI (6%). PE constituted 15% in RSR and 21% in FSR. PS and sphingomyelin were minor components of both SR (less than 4%). There were differences in the unsaturated chains of the total lipid extracts, PC, PE, and PI between FSR and RSR. RSR was high in linoleate and arachidonate while FSR contained substantial amounts of eicosapentaenoate and docosahexaenoate. FTIR spectroscopy revealed that the lipids of both membranes did not undergo a phase transition between 0 and 50 degrees C. The lipids were in the liquid-crystalline state at physiological temperatures and underwent monotonic increases in conformational disorder as the temperature was raised. CD spectra indicated higher content of alpha-helical structure of proteins in RSR than in FSR. Increasing temperature caused diminution of alpha-helix content. Relatively large decreases in ellipticity were observed between 20 degrees C and 40 degrees C for FSR and 30 degrees C and 60 degrees C for RSR. Measurements of intrinsic tryptophan fluorescence as a function of temperature gave similar results for membrane proteins in both FSR and RSR. The rate of change of tryptophan fluorescence and fluorescence lifetimes was constant over the temperature ranges studied, and no abrupt shifts in fluorescence occurred in the temperature regions where ellipticity decreased rapidly.
Animals↗
Levinthal's paradox.
Levinthal's paradox is that finding the native folded state of a protein by a random search among all possible configurations can take an enormously long time. Yet proteins can fold in seconds or less. Mathematical analysis of a simple model shows that a small and physically reasonable energy bias against locally unfavorable configurations, of the order of a few kT, can reduce Levinthal's time to a biologically significant size.
Mathematics↗
Reactivity to written mental arithmetic: effects of exercise lay-off and habituation.
The present study examined (i) the effects of exercise lay-off on heart rate (HR) and subjective response to mental stress in 24 individuals highly committed to exercise, and (ii) psychophysiological reactivity to a challenging written mental arithmetic with subjectively controlled difficulty level. Subjects were tested on two occasions one week apart. Exercise withdrawal did not influence psychophysiological stress response. Second exposure to the mental arithmetic resulted in significantly lower HR response, due to habituation; higher pretask resting HR, due to anticipation of performance; and later onset in HR recovery. No changes in task performance and subjective measures were observed from session one to session two, indicating that habituation is rather a physiological than behavioral phenomenon. While these findings do not strengthen the link between exercise and stress response, they demonstrate the significant mediatory roles of habituation and anticipation in laboratory studies employing a test-retest design.
Adult↗