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PubMed2006

Effects of quenched and annealed macromolecular crowding elements on a simple model for signaling in T lymphocytes.

Biochemical reactions in cells occur in an environment that is crowded in the sense that various macromolecular species and organelles occupy much of the space. The effects of molecular crowding on biochemical reactions have usually been studied in the past in a spatially homogeneous environment. However, signal transduction in cells is often initiated by the binding of receptors and ligands in two apposed cell membranes, and the pertinent biochemical reactions occur in a spatially inhomogeneous environment. We have studied the effects of crowding on biochemical reactions that involve both membrane proteins and cytosolic molecules by investigating a simplified version of signaling in T lymphocytes using a Monte Carlo algorithm. We find that, if signal transduction occurs on time scales that are slow compared to the motility of the molecules and organelles that constitute the crowding elements, the effects of crowding are qualitatively the same as in a homogeneous three-dimensional (3D) medium. In contrast, if signal transduction occurs on a time scale that is much faster than the time over which the crowding elements move, then the effects of varying the extent of crowding are very different when reactions occur in both 2- and 3D space. We discuss these differences and their origin. Since many signaling reactions are fast, our results may be useful for diverse situations in cell biology.

Binding Sites↗
PubMed2004

Tooth width ratios in crowded and noncrowded dentitions.

Discrepancies in tooth width ratios could affect the excellence in the finishing of orthodontic cases. This study compares tooth width ratios in crowded and noncrowded dental arches. Tooth widths were measured from 143 dental casts (40 crowded and 33 spaced in male individuals and 43 crowded and 27 spaced in female individuals). Simultaneous crowded or spaced arches were selected. Tooth width measurements were made with a sliding caliper with a Vernier scale neared 0.1 mm. Inter- (0.990) and intraexaminer (0.993) reliability with mean differences of 0.03 mm (-0.03; 0.07) and 0.01 mm (-0.04; 0.05), respectively, were attained. Bolton analysis was applied to calculate the tooth width ratios (anterior and total). Descriptive statistics, Student's t-test, and analysis of variance were applied. There was no significant difference between the anterior and total tooth width ratios according to sex (P > .05). Both anterior and total ratios were significantly greater in subjects with crowding (P = .003 and P = .026, respectively), but no statistical difference by arch type in anterior and total tooth mass excess was found (P > .05). Differences among subjects with noncrowded and crowded dentitions were of 0.39 and 0.51 mm for the excess of anterior and total upper tooth mass, respectively, with respect to lower mass excess. Although the anterior and overall ratios and the differences between the upper and lower tooth width sums are greater in subjects with crowding, no clinically significant difference is observed (less than one mm).

Adolescent↗
PubMed2005

The role of target salience in crowding.

We studied 'crowding' in the parafovea using orientation identification of a Gabor target as the task, and flanking Gabors on an isoeccentric circle as distractors. Orientation-discrimination thresholds were raised by nearby flanking distractors. This crowding effect was increased by the number of distractors and decreased by the spatial separation between target and distractors. Crowding was greatest when the target was in the centre of the distractor array and smallest when the target was on the edge of the array. A cue indicating the position of the target improved performance when the position was otherwise unknown and the spatial separation between target and distractors was large, but the cue had no significant effect when separation was small. Increasing the contrast of the target relative to the distractors reduced crowding, but targets of smaller contrast than that of the distractors are even harder to identify than those of the same contrast. Putting the target and distractors in different depth planes decreased crowding for some observers, but there were qualitative individual differences. A large (say, 45 degrees) difference in orientation between target and distractors caused the target to 'pop out' in a presence/absence task, despite the evidence from other studies that crowding is still found in these conditions. We conclude that salience has, at best, modest effects on crowding.

Cues↗
PubMed1999

Effects of macromolecular crowding on protein folding and aggregation.

We have studied the effects of polysaccharide and protein crowding agents on the refolding of oxidized and reduced hen lysozyme in order to test the prediction that association constants of interacting macromolecules in living cells are greatly increased by macromolecular crowding relative to their values in dilute solutions. We demonstrate that whereas refolding of oxidized lysozyme is hardly affected by crowding, correct refolding of the reduced protein is essentially abolished due to aggregation at high concentrations of crowding agents. The results show that the protein folding catalyst protein disulfide isomerase is particularly effective in preventing lysozyme aggregation under crowded conditions, suggesting that crowding enhances its chaperone activity. Our findings suggest that the effects of macromolecular crowding could have major implications for our understanding of how protein folding occurs inside cells.

Animals↗
PubMed2002

Effects of macromolecular crowding on protein folding and aggregation studied by density functional theory: dynamics.

Inside the living cell is inherently crowded with proteins and other macromolecules. Thus, it is indispensable to take into account various interactions between the protein and other macromolecules for thorough understanding of protein functions in cellular contexts. Here we focus on the excluded volume interaction imposed on the protein by surrounding macromolecules or "crowding agents." We have presented a theoretical framework for describing equilibrium properties of proteins in crowded solutions [A. R. Kinjo and S. Takada, Phys. Rev. E (to be published)]. In the present paper, we extend the theory to describe nonequilibrium properties of proteins in crowded solutions. Dynamics simulations exhibit qualitatively different morphologies depending on the aggregating conditions, and it was found that macromolecular crowding accelerates the onset of aggregation while stabilizing the native protein in the quasiuniform phase before the onset of aggregation. It is also observed, however, that the aggregation may be kinetically inhibited in highly crowded conditions. The effects of crowding on folding and unfolding of proteins are also examined, and the results suggest that fast folding is an important factor in preventing aggregation of denatured proteins.

Biophysical Phenomena↗
PubMed2005

Effect of macromolecular crowding agents on human immunodeficiency virus type 1 capsid protein assembly in vitro.

Previous studies on the self-assembly of capsid protein CA of human immunodeficiency virus type 1 (HIV-1) in vitro have provided important insights on the structure and assembly of the mature HIV-1 capsid. However, CA polymerization in vitro was previously observed to occur only at very high ionic strength. Here, we have analyzed the effects on CA assembly in vitro of adding unrelated, inert macromolecules (crowding agents), aimed at mimicking the crowded (very high macromolecular effective concentration) environment within the HIV-1 virion. Crowding agents induced fast and efficient polymerization of CA even at low (close to physiological) ionic strength. The hollow cylinders thus assembled were indistinguishable in shape and dimensions from those formed in dilute protein solutions at high ionic strength. However, two important differences were noted: (i) disassembly by dilution of the capsid-like particles was undetectable at very high ionic strength, but occurred rapidly at low ionic strength in the presence of a crowding agent, and (ii) a variant CA from a presumed infectious HIV-1 with mutations at the CA dimerization interface was unable to assemble at any ionic strength in the absence of a crowding agent; in contrast, this mutation allowed efficient assembly, even at low ionic strength, when a crowding agent was used. The use of a low ionic strength and inert macromolecules to mimic the crowded environment inside the HIV-1 virion may lead to a better in vitro evaluation of the effects of conditions, mutations or/and other molecules, including potential antiviral compounds, on HIV-1 capsid assembly, stability and disassembly.

Capsid Proteins↗
PubMed1996

The assessment of crowding without the need to record arch perimeter. Part I: Arches with acceptable alignment.

A simplified, mathematically determined technique for calculating arch perimeter (the overlap method) is described and its validity in determining an accurate assessment of crowding is tested. Indices, expressed in millimetres of crowding or spacing, were computed for both the whole arch and the labial segment alone. Study casts of 36 individuals with clinically acceptable lower arches were used to validate the method. The degree of crowding or spacing calculated was compared with a "clinical' assessment of each arch, in which the irregularity was measured directly using a steel ruler. Various calculation schemes were tested. Depending on exactly how the overlap was determined, the results varied slightly. Although the arches showed acceptable alignment, it was preferable to include a strategy for normalizing the positions of any rotated teeth before the overlaps were calculated. Repositioning any bucco-lingually displaced teeth into the line of the arch, however, was not useful. For the complete arch good agreement with the clinical assessment was reached on 31 occasions and for the labial segment, all but one appraisals were within 0.5 mm of each other. In the remaining instances (five complete arches and one labial segment), the degree of crowding or spacing was between 0.5 and 1 mm of the clinical assessment. Considering that clinical measurement of minor degrees of crowding and spacing cannot be precise, these results were considered acceptable. The method was easy to use, relying only on the recording of mesio-distal tooth widths and was acceptably reproducible. The technique would therefore appear to provide a valid yet simple research tool with which to record the degree of crowding. Its ability to cope with irregular and crowded arches will be the subject of a subsequent review.

Algorithms↗
PubMedDate not supplied

Effects of crowding of mice on humoral antibody formation and protection to lethal antigenic challenge.

The effects of grouping (crowding) on humoral antibody response to typhoid paratyphoid vaccine and subsequent protection from a minimal lethal challenge dose of Salmonella typhimurium were studied in white Swiss-Webster mice. The data show a trend between the degree of crowding and antibody response. Geometric mean titers of high density grouped mice were significantly lower than the geometric mean titers of the less crowded mice. Also, there were significantly less antibody responders in the high density grouped mice than in the less crowded mice. However, challenged with a minimal LD50 dose of Salmonella thphimurium, no deaths occurred in the immunized study group, regardless of measurable antibody level. In the nonimmunized controls, which were under the same stressor conditions, there was a significant difference between the level of crowding and death to challenge. The data show that nonimmunized mice in this study exhibited a marked increase in susceptibility to an infectious agent when under the stressor effect of crowding.

Animals↗
PubMed2003

Effect of cyclooxygenase inhibitors on the vasopressin induced ACTH and corticosterone response during crowding stress.

The aim of the present study was to compare the effect of social stress on the corticotropin releasing hormone (CRH) and arginine vasopressin (AVP)-induced pituitary-adrenocortical activity. Also the significance of prostaglandins (PG) generated by constitutive and inducible cyclooxygenase (COX-1 and COX-2) in the stimulation of hypothalamic-pituitary-adrenal (HPA) axis by AVP under basal and crowding stress conditions was investigated. The control rats were housed 7 in a standard cage and stressed rats were crowded 24 in a cage of the same size during 7 days. The activity of HPA axis was determined by measuring plasma ACTH and serum corticosterone levels 1 h after i.p. AVP administration. Indomethacin (2.0 mg/kg i.p.), a non-selective COX inhibitor, piroxicam (0.2, 2.0, and 5.0 mg/kg), a more potent COX-1 than COX-2 inhibitor, and compound NS-398 (0.2 and 2.0 mg/kg) a selective COX-2 inhibitor, were administered i.p. 15 min prior to AVP (5.0 microg/kg i.p.) to control or crowded rats. The obtained results indicate that social stress for 7 days considerably inhibits the stimulatory action of AVP on ACTH secretion, while it intensifies the CRH-induced ACTH secretion. Indomethacin, piroxicam and NS-398 significantly diminished the AVP-elicited ACTH and corticosterone secretion in non-stressed rats. None of these COX antagonist induced any significant inhibition of the AVP-induced ACTH and corticosterone secretion in stressed rats. Therefore, PG generated by COX-1 or COX-2 do not participate to a significant extent in the HPA stimulation by AVP during crowding stress. These results suggest that social crowding stress desensitizes the PG stimulatory mechanism which considerably mediates the AVP-induced HPA stimulation under basal conditions. The results contrast with a lack of any involvement of PG in the CRH-induced stimulation of HPA response under basal or crowding stress conditions.

Adrenocorticotropic Hormone↗
PubMed1979

Physiological, social, and behavioral consequences of crowding on children and adolescents.

The present investigation was designed to examine systematically the effects of short-term crowding, involving close physical proximity, on children and adolescents. Fourth-, eighth-, and eleventh-grade children participated in groups of 4 under high or moderate spatial density conditions. During exposure to these conditions, physiological responses were measured, and following this experience children engaged in a cooperation-competition activity and provided self-reports related to their participation. Results demonstrated that crowding had physiological, behavioral and social consequences for the children studied. Across all grade levels, males in particular were more affected by short-term crowding, displaying the highest elevations in stress-related arousal. Following exposure to the crowding conditions, children were more competitive, despite the fact that they had all to gain from cooperating and nothing to lose. Furthermore, children reported having felt crowded, tense, annoyed, and uncomfortable as a function of the close physical proximity to others. Implications of the immediate and residual effects of crowding on the behavior and functioning of children are discussed.

Adolescent↗
PubMedDate not supplied

Protein folding and binding in confined spaces and in crowded solutions.

Simple theoretical models are presented to illustrate the effects of spatial confinement and macromolecular crowding on the equilibria and rates of protein folding and binding. Confinement is expected to significantly stabilize the folded state, but for crowding only a marginal effect on protein stability is expected. In confinement the unfolded chain is restricted to a cage but in crowding the unfolded chain may explore different interstitial voids. Because confinement and crowding eliminate the more expanded conformations of the unfolded state, folding from the compact unfolded state is expected to speed up. Crowding will shift the binding equilibrium of proteins toward the bound state. The significant slowing down in protein diffusion by crowding, perhaps beneficial for chaperonin action, could result in a decrease in protein binding rates.

Models, Theoretical↗
PubMed1982

Incisor shape and incisor crowding: a re-evaluation of the Peck and Peck ratio.

Although lower incisor tooth shapes, defined as mesiodistal length divided by labiolingual width, have bee proposed as important factors in lower incisor crowding, the question of whether or not these ratios are more useful than simple measurements of incisor mesiodistal length has not been addressed. In order to test this question, we measured mesiodistal and labiolingual incisor dimensions and lower incisor crowding (defined as the crowding index proposed by Little) on dental casts from two groups: 100 pretreatment orthodontic patients and 100 Hutterites from a religious isolate in Canada. The orthodontic patients are our primary interest. The Hutterites serve mainly to test whether or not results are consistent in another population of different ethnicity, age distribution, and occlusal status. In each population, incisor crowding is correlated with the tooth shape ratios, confirming the general observations of Peck and Peck. However, mesiodistal incisor lengths have slightly higher correlations with crowding than the shape ratios. In multiple regression equations to predict crowding in each population, incisor mesiodistal lengths are the most important variable, and neither the tooth shape ratios nor labiolingual widths significantly improve the equations. Although statistically significant, none of the correlations is higher than 0.30, and they are thus of little clinical value. The use of tooth size measurements or ratios as a guide to clinical procedures is an oversimplification of a complex problem.

Canada↗
PubMed1983

An examination of dental crowding and its relationship to tooth size and arch dimension.

This investigation was undertaken to examine the extent to which tooth size and jaw size each contribute to dental crowding. Two groups of dental casts were selected on the basis of dental crowding. One group, consisting of 50 pairs of dental casts (18 males and 32 females), exhibited gross dental crowding. A second group, consisting of 54 pairs of dental casts (24 males and 30 females), exhibited little or no crowding. Means and standard deviations of the following parameters were used to compare the two groups: individual and collective mesiodistal tooth diameters, dental arch perimeters, and buccal and lingual dental arch widths. Statistically, the crowded and noncrowded groups could not be distinguished from each other on the basis of mesiodistal tooth diameters. However, significant differences were observed between the dental arch dimensions of the two groups. The crowded group was found to have smaller dental arch dimensions than the noncrowded group. The results of this study suggest that consideration be given to those treatment techniques which increase dental arch length rather than reduce tooth mass.

Adolescent↗
PubMedDate not supplied

Effect of crowd size on patient volume at a large, multipurpose, indoor stadium.

A prediction of patient volume expected at "mass gatherings" is desirable in order to provide optimal on-site emergency medical care. While several methods of predicting patient loads have been suggested, a reliable technique has not been established. This study examines the frequency of medical emergencies at the Syracuse University Carrier Dome, a 50,500-seat indoor stadium. Patient volume and level of care at collegiate basketball and football games as well as rock concerts, over a 7-year period were examined and tabulated. This information was analyzed using simple regression and nonparametric statistical methods to determine level of correlation between crowd size and patient volume. These analyses demonstrated no statistically significant increase in patient volume for increasing crowd size for basketball and football events. There was a small but statistically significant increase in patient volume for increasing crowd size for concerts. A comparison of similar crowd size for each of the three events showed that patient frequency is greatest for concerts and smallest for basketball. The study suggests that crowd size alone has only a minor influence on patient volume at any given event. Structuring medical services based solely on expected crowd size and not considering other influences such as event type and duration may give poor results.

Emergency Medical Services↗
PubMed2006

Crowding degrades saccadic search performance.

The identity of a target is more difficult to acquire when it is surrounded by distracters. The purpose of the present experiments was to investigate the implications of this crowding phenomenon for performance and eye movements in a real-life task as search with eye movements. The participants searched for a target in a one dimensional search strip. Above and below this search strip additional elements were added. In three conditions, the similarity of these mask elements to the search elements was varied. The spatial extent of crowding is known to increase with target-mask similarity [Nazir, T. A. (1992). Effects of lateral masking and spatial precueing on gap-resolution in central and peripheral vision. Vision Research, 32, 771-777, Kooi, F. L., Toet, A., Tripathy, S. P., & Levi, D. M. (1994). The effect of similarity and duration on spatial interaction in peripheral vision. Spatial Vision, 8(2), 255-279]. One condition did not contain masks. In a visibility experiment, we firstly validated this crowding manipulation. In the search experiment, we subsequently found that with increasing crowding search times were up to 76% longer. Eye movements were also affected. The number of fixations and fixation duration increased and saccade amplitude decreased with increasing crowding. We conclude that in order to understand eye movements in (everyday) tasks that require active exploration of the visual scene, crowding should be taken into account.

Adult↗
PubMed2006

Acuity, crowding, reading and fixation stability.

People with age-related macular disease frequently experience reading difficulty that could be attributed to poor acuity, elevated crowding or unstable fixation associated with peripheral visual field dependence. We examine how the size, location, spacing and instability of retinal images affect the visibility of letters and words at different eccentricities. Fixation instability was simulated in normally sighted observers by randomly jittering single or crowded letters or words along a circular arc of fixed eccentricity. Visual performance was assessed at different levels of instability with forced choice measurements of acuity, crowding and reading speed in a rapid serial visual presentation paradigm. In the periphery: (1) acuity declined; (2) crowding increased for acuity- and eccentricity-corrected targets; and (3), the rate of reading fell with acuity-, crowding- and eccentricity-corrected targets. Acuity and crowding were unaffected by even high levels of image instability. However, reading speed decreased with image instability, even though the visibility of the component letters was unaffected. The results show that reading performance cannot be standardised across the visual field by correcting the size, spacing and eccentricity of letters or words. The results suggest that unstable fixation may contribute to reading difficulties in people with low vision and therefore that rehabilitation may benefit from fixation training.

Adult↗
PubMed2004

Duplex dissociation of telomere DNAs induced by molecular crowding.

Because of the importance of telomere DNAs, the structures of these DNAs in vivo are currently of great research interest in the medical, pharmaceutical, chemical, and industrial fields. To understand the structure of biomolecules in vivo, their properties studied in vitro are extrapolated to the in vivo condition, while the condition in a living cell is inherently molecularly crowded and a nonideal solution contains various biomolecules. We investigated the effect of molecular crowding, which is one of the most important cellular environmental conditions, on the structure and stability of the telomere and G-rich and C-rich DNAs using circular dichroism (CD) spectra, CD melting curves, and isothermal titration calorimetry (ITC). The CD spectra and CD melting curves of G-rich DNA, C-rich DNA, and the 1:1 mixture of G-rich and C-rich DNAs showed that each G-rich DNA, C-rich DNA, and the 1:1 mixture form the antiparallel G-quadruplex, I-motif, and duplex, respectively, in the noncrowding condition as previously considered. On the contrary, the G-rich and C-rich DNAs individually form the parallel G-quadruplex and I-motif, respectively, in the molecular crowding condition, and the 1:1 mixture folds into the parallel G-quadruplex and I-motif but does not form a duplex. The ITC measurements indicated that the thermodynamic stability (DeltaG degrees (20)) of the duplex formation between the G-rich and C-rich DNAs in the noncrowding condition was -10.2 kcal mol(-)(1), while only a small heat change was observed in the ITC measurements in the molecular crowding condition. These ITC results also demonstrated that the molecular crowding condition prevents any duplex formation between G-rich and C-rich DNAs. These results indicate that a structural polymorphism of the telomere DNAs is induced by molecular crowding in vivo.

Calorimetry↗
PubMed2004

Malocclusion and crowding in an orthodontically referred Turkish population.

This study was conducted to evaluate malocclusion and crowding in 1356 patients (793 girls, 563 boys) referred to the Department of Orthodontics, Suleyman Demirel University, Turkey. Class I was the most frequently seen malocclusion in this referred Turkish orthodontic population whereas Class II, division 2 was the least frequently seen. Comparison of mean ages of the malocclusion groups indicated statistically significant difference between Class I and Class II, division 1 groups (P < .05). The lowest mean age was present in the Class II, division 1 group. Mild mandibular crowding was the most common finding whereas severe mandibular crowding was seen least frequently in all malocclusion groups. Cross tabulation of maxillary and mandibular crowding indicated that mild maxillary and severe mandibular crowding in the same patient was rarely seen in all types of malocclusions. Moderate maxillary and severe mandibular crowding in the same patient was another rare finding for all malocclusion groups.

Adolescent↗