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PubMed2004

Crowding under scotopic conditions.

Under certain circumstances, a subject's ability to discriminate spatial features of a target may be hampered by neighbouring contours. This phenomenon is popularly known as the "crowding effect", and it has been intensely studied for photopic vision: little attention has been paid to the effect at lower light levels. The underlying basis of the crowding effect has recently provoked some conjecture, with Hess and colleagues claiming that a passive "physical" phenomenon may either wholly [Vis. Res. 40 (2000) 365], or partially [J. Opt. Soc. Am. A--Opt. Image Sci. Vis. 17 (2000) 1516], account for the effect. In order to investigate the crowding effect under scotopic conditions, we conducted scotopic frequency of seeing experiments for Landolt C targets presented both with, and without, flanking bars; the size of the targets was varied so that frequency of seeing curves could be derived for each stimulus condition. Our results suggest that the spatial extent of crowding is significantly less for scotopic vision than for photopic vision at the same eccentricity--furthermore the effect does not seem to scale in proportion to target size. We also compared the resulting empirical curves to those that would be predicted by the hypothesis of Hess and colleagues. Our results do not support the hypothesis that the scotopic crowding effect is caused by a passive physical process.

Dark Adaptation↗
PubMed2004

Adaptation to apparent motion in crowding condition.

Visual adaptation has been successfully used for studying the neural activity of different cortical areas in response to visual stimuli when observers do not have explicit conscious access to those stimuli. We compared the orientation selective adaptation to apparent motion and its effect on the perception of stimuli with bistable apparent motion in crowded and non-crowded conditions. In the crowding paradigm conscious access to a visual stimulus is severely impaired when it is flanked by other similar stimuli in the peripheral visual field. As expected, adaptation to the target stimulus occurred in the non-crowded condition in all of the individual subjects (n=4; P<0.001). Although in the crowded condition subjects were not able to discriminate the target stimulus, adaptation to that stimulus was still preserved (P<0.001). There was no significant difference between the adaptations in the two conditions of the apparent motion (P>0.05). Imaging studies have shown that V5 cortex is the earliest visual area that specifically responds to apparent motion. Our results suggest that in certain conditions V5 may be activated while there is no explicit conscious access to the apparent motion.

Adaptation, Psychological↗
PubMed2003

Competition between protein folding and aggregation with molecular chaperones in crowded solutions: insight from mesoscopic simulations.

The living cell is inherently crowded with proteins and macromolecules. To avoid aggregation of denatured proteins in the living cell, molecular chaperones play important roles. Here we introduce a simple model to describe crowded protein solutions with chaperone-like species based on a dynamic density functional theory. As predicted by others, our simulations show that macromolecular crowding enhances the association of proteins and chaperones. However, when the intrinsic folding rate of the protein is slow, it is possible that crowding also enhances aggregation of proteins. The results of simulation suggest that, when the concentration of the crowding agent is as high as that in the cell, the association of the protein and unbound chaperone becomes correlated with the aggregation process, and that the protein-bound chaperones efficiently destroy the potential nuclei of aggregates and thus prevent the aggregation.

Biophysics↗
PubMed2002

Crowding effects on word identification in developmental dyslexia.

The effect of crowding on the identification of words was examined in normal readers and subjects with developmental dyslexia. In Experiment 1, a matching task was used. Words were presented either alone or embedded in other words. Vocal reaction times (RT) of dyslexics were slower and more sensitive to the presence of the surrounding stimuli than those of control subjects. Similar results were obtained in a control experiment using the same task for strings of symbols (isolated or crowded) instead of words. These data indicate that differences in crowding in control and dyslexic subjects arise at a pre-linguistic level. In Experiment 2, vocal RTs to word reading were measured. Two conditions putatively reducing the effect of crowding were tested: increasing inter-letter spacing and blurring. A moderate increase of inter-letter spacing produced faster vocal RTs in dyslexics, while no effect was present in normal controls. Moderate blurring of stimuli did not change dyslexics' RTs, while normal readers became slower. Group and individual results are discussed to evaluate the extent to which crowding contributes to the genesis of developmental dyslexia.

Adolescent↗
PubMed2001

Modelling host cell availability and the crowding effect in Eimeria infections.

Within-host mathematical models of Eimeria maxima and Eimeria praecox infections of the chicken are presented and used to investigate the role of host cell availability as a possible determinant of the so-called 'crowding effect'; whereby the fecundity of the parasites decreases as infectious dose increases. Assumptions about the number of available host cells, the average lifespan of these cells and the age structure within the host-cell population were made and mathematical models were constructed and combined with experimental data to test whether these conditions could reproduce the crowding effect in the two species. Experimental data demonstrated that crowding during in vivo infections was apparent following very low infectious doses, but none of the models could adequately reproduce crowding at the same doses while maintaining realistic estimates of the dynamics of the enterocyte pool. However, both the size and lifespan of the enterocyte pool were demonstrated to have substantial effects on the fecundity of the infections, particularly at higher doses. These data indicate that host cell availability cannot be solely responsible for the crowding effect. Alternative factors such as the influence of the primary immune response to the parasite may also be explored using within-host models and other applications of these models are discussed.

Animals↗
PubMed2001

Can the amplitude difference spectrum peak frequency explain the foveal crowding effect?

Hess et al. (Vision Res. 40 (2000) 365) found that the peak frequency of the amplitude difference spectrum (ADS) of a Landolt C/flanking bar configuration was higher than that of an isolated C. They believed that this mismatch in the dominant frequency components could account for the foveal crowding effect. The following empirical and theoretical studies were conducted to evaluate this new explanation. First, the foveal crowding effect between a Landolt C and four flanking bars was measured under same- and mixed-polarity conditions. Significant crowding was found under the mixed-polarity condition. This result is different from the data that Hess et al. used to support their explanation of the foveal crowding effect. Second, analytical expressions for the Fourier transforms of C/flanking bar configurations were derived. ADS for various ring/bar separations and contrast polarities were calculated using these expressions. This analysis showed several discrepancies between ADS peak frequency predictions and empirical data. Therefore, the ADS peak frequency of the stimulus configuration does not provide an adequate explanation for the foveal crowding effect.

Fourier Analysis↗
PubMed2000

The use of the lingual arch in the mixed dentition to resolve incisor crowding.

In the mixed dentition, arch length preservation, maintaining the leeway space, can often provide adequate space to resolve lower incisor crowding. Yet the frequency of this occurrence is not known. To obtain this information, lingual arches were placed in the mandibular arches of 107 consecutive mixed dentition patients with incisor crowding to preserve arch length and make the leeway space available to resolve the crowding. Arch length decreased only 0.44 mm whereas the intercanine, interpremolar, and intermolar dimensions increased between 0.72 and 2.27 mm. There was adequate space to resolve the crowding in 65 (60%) of the 107 patients. If perfect arch length preservation had occurred, there would have been adequate space to resolve the crowding in 73 (68%) of the 107 patients. The correlation between leeway space and tooth size-arch size discrepancy was only 0.44.

Bicuspid↗
PubMed1990

Late lower arch crowding: the effect of second molar extraction.

Mesial drift and change in crowding over a 5-year period were compared in 30 subjects whose lower second molars were extracted between ages 11 and 17 years and 30 subjects whose lower second molars were not extracted. Significant differences were found. The nonextraction subjects showed an average forward movement of first molars and an increase in crowding. The extraction group showed slight distal movement of first molars and a decrease in crowding. It is concluded that lower second molar extraction reduces the tendency to forward movement of buccal segments and increased crowding by relieving eruptive pressure from third molars and that the presence of a developing lower third molar with insufficient space can be one cause of late lower arch crowding.

Adolescent↗
PubMed2001

Macromolecular crowding: obvious but underappreciated.

Biological macromolecules evolve and function within intracellular environments that are crowded with other macromolecules. Crowding results in surprisingly large quantitative effects on both the rates and the equilibria of interactions involving macromolecules, but such interactions are commonly studied outside the cell in uncrowded buffers. The addition of high concentrations of natural and synthetic macromolecules to such buffers enables crowding to be mimicked in vitro, and should be encouraged as a routine variable to study. The stimulation of protein aggregation by crowding might account for the existence of molecular chaperones that combat this effect. Positive results of crowding include enhancing the collapse of polypeptide chains into functional proteins, the assembly of oligomeric structures and the efficiency of action of some molecular chaperones and metabolic pathways.

Animals↗
PubMed2005

Methyl displacements from cyclopentadienyl ring planes in sterically crowded (C5Me5)3M complexes.

The displacements of the methyl substituents away from the metal and out of the cyclopentadienyl ring plane are compared in sterically crowded (C(5)Me(5))(3)M complexes vs sterically normal f-element complexes in an attempt to evaluate the utility of this parameter in predicting unusual (C(5)Me(5))(1-) ring reactivity. The out-of-plane displacements of 16 sterically crowded tris(cyclopentadienyl) complexes of general formula (C(5)Me(5))(3)M, (C(5)Me(4)R)(3)M (R = Et, (i)Pr, (t)()Bu, SiMe(3)), (C(5)Me(5))(3)MX (X = anion), and (C(5)Me(5))(3)ML (L = neutral ligand) are compared with [(C(5)Me(5))(2)U](2)(C(6)H(6)), (C(5)Me(5))(2)Sm(PC(4)H(2)(t)Bu(2)), and 33 representative examples of f-element bis(cyclopentadienyl) complexes with normal cyclopentadienyl behavior and coordination numbers ranging from 6 to 10. In general, the methyl displacement values of sterically crowded complexes overlap with those in the other complexes, which demonstrates that the basis of the structural distortions is complex. However, if the most extreme out-of-plane displacement in each of the sterically crowded complexes is examined vs the analogous maximum out-of-plane displacement in less crowded systems, there appears to be a basis for predicting cyclopentadienyl reactivity.

Journal Article↗
PubMed2006

Hydration regulates thermodynamics of G-quadruplex formation under molecular crowding conditions.

The effect of molecular crowding on the structure and stability of biomolecules has become a subject of increasing interest because it can clarify how biomolecules behave under cell-mimicking conditions. Here, we quantitatively analyzed the effects of molecular crowding on the thermodynamics of antiparallel G-quadruplex formation via Hoogsteen base pairs and of antiparallel hairpin-looped duplex (HP duplex) formation via Watson-Crick base pairs. The free energy change at 25 degrees C for G-quadruplex formation decreased from -3.5 to -5.5 kcal mol(-1) when the concentration of poly(ethylene glycol) 200 was increased from 0 to 40 wt %, whereas that of duplex formation increased from -9.8 to -6.9 kcal mol(-1). These results showed that the antiparallel G-quadruplex is stabilized under molecular crowding conditions, but that the HP duplex is destabilized. Moreover, plots of stability (ln K(obs)) of the DNA structures versus water activity (ln a(w)) demonstrated that the ln K(obs) for G-quadruplex formation decreased linearly as the ln a(w) increased, whereas that for duplex formation increased linearly with the increase in ln a(w), suggesting that the slope approximately equals the number of water molecules released or taken up during the formation of these structures. Thus, molecular crowding affects the thermodynamics of DNA structure formation by altering the hydration of the DNA. The stabilization of the DNA structures with Hoogsteen base pairs and destabilization of DNA structures with Watson-Crick base pairs under molecular crowding conditions lead to structural polymorphism of DNA sequences regulated by the state of hydration.

Base Pairing↗
PubMed1995

Variable effects of crowding on Drosophila hosts of male-lethal and non-male-lethal spiroplasmas in laboratory populations.

Male-lethal, maternally inherited spiroplasmas occur in four species of Drosophila, and persist in natural populations despite imperfect vertical transmission rates. In the field, larval crowding is thought to be sporadic, but occasionally intense. To determine whether crowding affects host persistence, I compared the population dynamics of infected females (hosts) under crowded conditions to those expected from data collected on uncrowded females. I estimated host fitness components and maternal transmission rates for individual females under uncrowded conditions in both the artificial host D. pseudoobscura (this paper) and the native host D. willistoni (previously reported). Spiroplasma infection had no effect on lifetime production of daughters in D. pseudoobscura; however, as with some D. willistoni lines, hosts may produce more of their daughters earlier in life than nonhosts. Because individual contributions to relative rates of increase calculated from these fitness data were similar for hosts and nonhosts, I expected hosts to persist in laboratory populations. Instead, three patterns were observed: rapid extinction of D. willistoni females infected with male-lethal spiroplasmas, slow decline or persistence of hosts (depending on initial frequency) in both D. pseudoobscura infected with male-lethal spiroplasmas, and D. willistoni infected with non-male-lethal spiroplasmas. Population dynamics, then, depend on host species and bacterial isolate. Fitness estimates change with host line in uncrowded D. willistoni, but host genetic background did not affect population dynamics. These and previously published results show that the interaction phenotype changes with host and parasite isolate, and that host fitness can be affected by crowding. Crowding in natural populations may therefore decrease host fitness but, in expanding populations, early reproduction in hosts may be to their advantage. Possible effects of seasonal fluctuations in population density on the fitness of infected Drosophila are discussed.

Animals↗
PubMed1998

Implications of macromolecular crowding for signal transduction and metabolite channeling.

The effect of different total enzyme concentrations on the flux through the bacterial phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) in vitro was determined by measuring PTS-mediated carbohydrate phosphorylation at different dilutions of cell-free extract of Escherichia coli. The dependence of the flux on the protein concentration was more than linear but less than quadratic. The combined flux-response coefficient of the four enzymes constituting the glucose PTS decreased slightly from values of approximately 1.8 with increasing protein concentrations in the assay. Addition of the macromolecular crowding agents polyethylene glycol (PEG) 6000 and PEG 35000 led to a sharper decrease in the combined flux-response coefficient, in one case to values of approximately 1. PEG 6000 stimulated the PTS flux at lower protein concentrations and inhibited the flux at higher protein concentrations, with the transition depending on the PEG 6000 concentration. This suggests that macromolecular crowding decreases the dissociation rate constants of enzyme complexes. High concentrations of the microsolute glycerol did not affect the combined flux-response coefficient. The data could be explained with a kinetic model of macromolecular crowding in a two-enzyme group-transfer pathway. Our results suggest that, because of the crowded environment in the cell, the different PTS enzymes form complexes that live long on the time-scale of their turnover. The implications for the metabolic behavior and control properties of the PTS, and for the effect of macromolecular crowding on nonequilibrium processes, are discussed.

Cell-Free System↗
PubMed2001

Effects of macromolecular crowding on the refolding of glucose- 6-phosphate dehydrogenase and protein disulfide isomerase.

The effects of polysaccharide, polyethylene glycol, and protein-crowding agents on the refolding of glucose-6-phosphate dehydrogenase (G6PDH) and protein disulfide isomerase have been examined. By increasing concentration during refolding, the reactivation yields of the two proteins decrease with the formation of soluble aggregates. In the presence of high concentrations of crowding agents the reactivation yields remain constant but with decreased refolding rates. The refolding of G6PDH changes from monophasic to biphasic first-order reactions in the presence of crowding agents, and the amplitude of the new slow phase increases with increasing concentrations of crowding agents. The molecular chaperone GroEL reverses the refolding kinetics of G6PDH from biphase back to monophase and accelerates the refolding process. Our results display the complexity and diversity of the effects of macromolecular crowding on both the thermodynamics and kinetics of protein folding.

Chaperonin 60↗
PubMed2002

Volume exclusion effect as a driving force for reverse proteolysis. Implications for polypeptide assemblage in a macromolecular crowded milieu.

Macromolecular crowding, in principle, should affect any reaction that is accompanied by significant reduction in excluded volume. Here we have examined the influence of crowding on reverse proteolysis. We show that proteosynthesis of a polypeptide product with an interacting folding motif such as coiled coil is facilitated in crowded media as a consequence of the volume exclusion effect. Further, we demonstrate that crowding could also effect the conversion of a noncovalent protein complex (fragment complementing protein) obtained by limited proteolysis to the native covalent form, but only if the formation of the native protein results in large compaction leading to a substantial volume exclusion effect. Subtilisin-catalyzed reformation of native triosephosphate isomerase (TIM) from multiple fragments is facilitated by crowding. However, a single nick in ribonuclease S (RNase S) could not be ligated under similar conditions. The failure of generation of RNase A from RNase S is consistent with the fact that the crystal structure of the two forms are almost superimposable, and hence no significant difference of volume exclusion exists between reactant (RNase S) and product (RNase A). In contrast, considerable compaction, and consequently large reduction in excluded volume, is attained through the assembly of a TIM barrel structure. Taken together, these results have implications for both in vitro as well as in vivo polypeptide assemblage by reverse proteolysis.

Amino Acid Sequence↗
PubMed2004

Dental crowding in a prehistoric population.

The aims of this study were to investigate dental crowding from the Copper Age and examine the extent and patterns of wear. Crowding was estimated in 43 adult mandibles using Little's irregularity index. Dental wear, dental diameters, arch width and the presence of third molars were also studied as possible aetiological factors for crowding. The remains were found at the archaeological site of Roaix, located in the south of France. Radiocarbon dating indicated that the lower layer was from 2150 +/- 140 years BC (date +/- 1 standard deviation) and the upper level from 2090 +/- 140 years. The graves were estimated to contain the remains of 150 adults and 50 children. Forty-three intact mandibles were used for this study. All of the mandibles presented incisor crowding with a majority of minimal and moderate irregularities, but in seven cases there were extreme irregularities and in two canine impaction was observed. These results are in contrast with the literature where it is reported that malocclusions were rare in prehistoric populations. The findings of this study suggest that crowding may be of a genetic origin and might not be caused by excessive tooth size or changes in environmental factors (masticatory activity).

Adolescent↗
PubMed2004

Protein folding by the effects of macromolecular crowding.

Unfolded states of ribonuclease A were used to investigate the effects of macromolecular crowding on macromolecular compactness and protein folding. The extent of protein folding and compactness were measured by circular dichroism spectroscopy, fluorescence correlation spectroscopy, and NMR spectroscopy in the presence of polyethylene glycol (PEG) or Ficoll as the crowding agent. The unfolded state of RNase A in a 2.4 M urea solution at pH 3.0 became native in conformation and compactness by the addition of 35% PEG 20000 or Ficoll 70. In addition, the effects of macromolecular crowding on inert macromolecule compactness were investigated by fluorescence correlation spectroscopy using Fluorescence-labeled PEG as a test macromolecule. The size of Fluorescence-labeled PEG decreased remarkably with an increase in the concentration of PEG 20000 or Ficoll 70. These results show that macromolecules are favored compact conformations in the presence of a high concentration of macromolecules and indicate the importance of a crowded environment for the folding and stabilization of globular proteins. Furthermore, the magnitude of the effects on macromolecular crowding by the different sizes of background molecules was investigated. RNase A and Fluorescence-labeled PEG did not become compact, and had folded conformation by the addition of PEG 200. The effect of the chemical potential on the compaction of a test molecule in relation to the relative sizes of the test and background molecules is also discussed.

Animals↗
PubMed1997

Cortisol and glucose responses after acute stress by net handling in the sparid red porgy previously subjected to crowding stress

In red porgy Pagrus pagrus subjected to 3 weeks of chronic stress by crowding, plasma cortisol remained significantly higher in crowded fish compared to controls. There was no significant effect of crowding on plasma glucose levels. When the crowded fish were subjected to an acute handling the plasma cortisol response was similar to that of the uncrowded fish. No significant differences were found between the groups. The changes in plasma glucose following acute handling were also similar in both crowded and uncrowded fish.

Journal Article↗