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PubMed2000

Household crowding and compliance with outpatient treatment in patients with non-affective functional psychoses in São Paulo, Brazil.

BACKGROUND: Compliance with outpatient treatment can reduce the use of psychiatric inpatient services by people with severe mental disorders. In developing countries, socio-economic factors may be associated with compliance with outpatient treatment. METHODS: A 2-year prospective cohort study was conducted of 99 patients with non-affective functional psychoses who were discharged from hospital. Standardized assessments were used for psychopathology, social functioning and use of psychiatric services. RESULTS: Forty-two patients (42.4%) missed all outpatient appointments for at least 2 consecutive months. Household crowding was the only variable associated with poor compliance, patients living in very crowded homes being more than twice as likely to show poor compliance as those living in less crowded homes. CONCLUSIONS: In large urban centres in developing countries, strategies to improve compliance with outpatient treatment targeted towards those living in overcrowded households may reduce use of psychiatric beds.

Adolescent↗
PubMed2006

On the origins of a crowded cytoplasm.

Contemporary cells show a highly crowded macromolecular content, the processes which originated this state being largely unknown. We propose that a driving force leading to the crowded cellular state could be the increase in growth rate produced by an enhanced cytoplasmic protein concentration. Briefly, in a diluted scenario, an increase in protein concentration has two opposing effects on growth rate. The favorable effect is the increase in the activity per unit volume of the component proteins and the disadvantageous effect is the concomitant increase in the protein mass per unit volume which has to be produced. In this work we show that the first effect is quantitatively more important, resulting in an overall increase in growth rate. This result was obtained with a model of E. coli and using nonmechanistic physiological arguments. The proposed driving force operates even at low protein concentrations, where the nonspecific interactions of macromolecular crowding are not significant, and could be as ancient as the first protocells. Experimental measurement of this cytoplasmic protein concentration effect in present organisms is hindered by the prevailing nonspecific interactions, product of long-term evolution. However, chemical/biochemical systems, built up to mimic properties of living cells, could be an adequate tool to test this effect.

Adaptation, Physiological↗
PubMed2004

Perceptual salience of global structures and the crowding effect in amblyopia.

BACKGROUND: The crowding effect refers to stronger deficits in linear acuity (e.g., letters in a line) than in single letter acuity in amblyopia. The current work investigated whether the salience of a global structure in which the target for identification is embedded influences the crowding effect in amblyopia. METHODS: Compound shapes were presented to the amblyopic and fellow eyes respectively of 12 anisometropic amblyopes. The compound stimuli were presented on either a blank or a cross background so that the salience of global structures were manipulated. Reaction times (RTs) and response error rates were recorded when subjects identified global or local shapes, respectively. RESULTS: RTs were shorter to global than local shapes for both the amblyopic and the fellow eyes. The global RT advantage was larger for the amblyopic than the fellow eye. Interestingly, when viewing the stimuli with the amblyopic eye, subjects made more errors to local targets when the compound stimuli were presented against the blank than the cross background. CONCLUSION: The results suggest that the salience of global structures of visual stimuli contributes to the crowding effect in amblyopia.

Adolescent↗
PubMed1982

Extraction of third molars in cases of anticipated crowding in the lower jaw.

The purpose of this study was to ascertain whether the lower third molar, in combination with other variables such as facial morphology and space conditions, can contribute to the occurrence or aggravation of crowding. The subjects consisted of twenty-three boys and twenty-nine girls with impacted third molars on both sides of the mandible. The impacted molar on one side was removed, while the other, nonextracted side was used as a control. Average age at the time of operation was 15.5 years (range, 13 to 19). Close to the operation and annually for at least 3 years afterward, study casts and cephalograms (lateral, frontal, and oblique) were taken. The findings indicated that (1) despite analyses of many variables, this study has not been able to predict which patients should react favorably or unfavorably to removal of the third lower molars in cases of anticipated crowding; (2) in cases with severe crowding removal of the molars could be recommended; (3) correct proximal contacts seem to be of importance in keeping the space that is achieved by extraction, while incorrect ones may spoil it.

Adolescent↗
PubMed1988

Macromolecular crowding extends the range of conditions under which DNA polymerase is functional.

The nick-translation reaction of E. coli DNA polymerase I (Pol I) was used as a model system to demonstrate the ability of macromolecular crowding to alter the response of an enzyme to a number of basic parameters, such as pH, temperature or inhibitors. In the presence of high concentrations of non-specific polymers, nick translation occurred under a variety of otherwise strongly inhibitory conditions. The conditions tested included a range of pH values or temperatures or inhibitory concentrations of urea, formamide or ethidium bromide. These crowding effects are accentuated at higher ionic strengths, suggesting their origin in increased binding between the polymerase and its DNA template-primer under crowded conditions. Kinetic measurements were consistent with such a mechanism.

DNA↗
PubMed1994

Macromolecular crowding effects on the interaction of DNA with Escherichia coli DNA-binding proteins: a model for bacterial nucleoid stabilization.

DNA-binding protein fractions from exponential and stationary phase cell extracts of E. coli were isolated by affinity chromatography on native DNA-cellulose. The ability of these fractions to convert DNA into a readily-sedimented form was compared in the absence or presence of added polymers. In the absence of polymers, large amounts of the proteins were required. In the presence of polyethylene glycol or polyvinylpyrrolidone, much smaller amounts of the DNA-binding proteins were required, indicating a macromolecular crowding effect from these polymers. The enhanced binding under crowded conditions appears to resolve a paradox between the cellular abundance of the DNA-binding proteins and the amounts required in earlier in vitro studies. The 'histone-like' protein HU from the DNA-binding protein fraction was preferentially incorporated into the pelleted DNA in the presence of polymers. Purified HU at roughly similar amounts caused a similar conversion of DNA to a readily-sedimentable ('condensed') form. Crowding-enhancement of DNA condensation by promoting the binding of proteins to the DNA provides a model for the stabilization of systems such as the bacterial nucleoid or kinetoplast DNA.

Bacterial Proteins↗
PubMed1986

Prevalence of tooth agenesis correlated with jaw relationship and dental crowding.

Two groups of patients were selected from the State University Dental School in Ghent--an experimental group with tooth agenesis and a control group with complete dentitions. Skeletal anomalies were diagnosed by means of lateral cephalograms according to the Sassouni analysis. The amount of crowding was measured on standardized photographs by Little's Irregularity Index. The prevalence of Class I skeletal relationship appeared to be significantly higher in the agenesis group than in the control group. Skeletal deep- and normal-bite cases occurred more often in patients with hypodontia than in the control group. As far as crowding was concerned, it was less pronounced in the hypodontia group (lateral segments) than in the control group. This conclusion held true only when the "amount" of crowding was not taken into account.

Adolescent↗
PubMed2006

Molecular crowding increases the amplification success of multiple displacement amplification and short tandem repeat genotyping.

Multiple displacement amplification (MDA) can generate large quantities of genomic DNA product from small amounts of template. We investigated the ability of MDA to amplify samples containing very small amounts of target DNA (5 pg to 1 ng) in the presence of a second, larger DNA sample for downstream short tandem repeat (STR) multiplex genotyping. We observed that STR amplification success of the minor fraction was increased in these mixed samples when compared with standard PCR only or MDA containing only the single trace DNA sample. Increased numbers of alleles were detected, with less amplification bias between loci than in single source samples undergoing the same protocol. To improve the STR genotyping accuracy, animal DNA was substituted for the additional human DNA, maintaining the increase in the number and quality of human-specific STR loci amplified. Polyethylene glycol 400, a commonly used crowding agent, was used as a replacement for the added genomic DNA in the MDA reaction and produced very similar results. Therefore, we suggest that additional DNA is acting as a molecular crowding agent during MDA. Performing MDA on trace amounts of DNA under crowded conditions results in greater numbers of alleles being amplified and more balanced amplification occurring between alleles.

Alleles↗
PubMed2004

Internal organisation of the nucleus: assembly of compartments by macromolecular crowding and the nuclear matrix model.

Many and possibly all macromolecules in the nucleus are segregated into discrete compartments, but the current model that this is achieved by a fibrillar nuclear matrix which structures the nuclear interior and compartments is not consistent with all experimental observations, as reviewed here. New results are presented which suggest that macromolecular crowding forces play a crucial role in the assembly of at least two compartments, nucleoli and PML bodies, and an in vitro system in which crowding assembles macromolecular complexes into structures which resemble nuclear compartments is described. Crowding forces, which are strong in the nucleus due to the high macromolecule concentration (in the range of 100 mg/ml), vastly increase the association constants of intermolecular interactions and can segregate different macromolecules into discrete phases. The model that they play a role in compartmentalisation of the nucleus is generally consistent with the properties of compartments, including their spherical or quasispherical form and their dynamic and mobile nature.

Animals↗
PubMed2006

Phase-related body-color polyphenism in hatchlings of the desert locust, Schistocerca gregaria: re-examination of the maternal and crowding effects.

The mechanism controlling the body color of hatchlings was studied for the desert locust, Schistocerca gregaria. A pheromonal factor secreted by gregarious female adults into the foam plugs of egg pods has been suggested to cause darkening in their progeny. We re-examined the role of this maternal factor by washing or separating eggs at deposition. Eggs produced by crowd-reared female adults were washed with saline or separated individually without being washed immediately after deposition and the body color of the hatchlings from them was compared with that from the eggs unwashed and kept in the egg pod until hatching. Most hatchlings were dark and no significant difference was found in the proportions of dark- and light-colored hatchlings between the treatments and controls. Likewise, eggs separated before the foam plug deposition produced dark-colored hatchlings as in the un-separated controls. These results demonstrated that neither washing nor separation of eggs at deposition affected the hatchling body coloration. The variation in hatchling body color was correlated closely to the body weight at hatching, indicating that hatchling body color had been determined maternally. Green hatchlings reared under crowded conditions remained green until the second stadium at which black patterns were induced. It was concluded that body color at hatching has been determined maternally and crowding during the first nymphal stadium influences nymphal body color but its effect is not manifested until the second stadium. The present study casts doubts on the presence of a recently suggested pheromonal factor on the color of the hatchlings.

Animals↗
PubMed2005

Effects of pH, salt, and macromolecular crowding on the stability of FK506-binding protein: an integrated experimental and theoretical study.

Environmental variables can exert significant influences on the folding stability of a protein, and elucidating these influences provides insight on the determinants of protein stability. Here, experimental data on the stability of FKBP12 are reported for the effects of three environmental variables: pH, salt, and macromolecular crowding. In the pH range of 5-9, contribution to the pH dependence of the unfolding free energy from residual charge-charge interactions in the unfolded state was found to be negligible. The negligible contribution was attributed to the lack of sequentially nearest neighboring charged residues around groups that titrate in the pH range. KCl lowered the stability of FKBP12 and the E31Q/D32N double mutant at small salt concentrations but raised stability after approximately 0.5 M salt. Such a turnover behavior was accounted for by the balance of two opposing types of protein-salt interactions: the Debye-Hückel type, modeling the response of the ions to protein charges, favors the unfolded state while the Kirkwood type, accounting for the disadvantage of the ions moving toward the low-dielectric protein cavity from the bulk solvent, disfavors the unfolded state. Ficoll 70 as a crowding agent was found to have a modest effect on protein stability, in qualitative agreement with a simple model suggesting that the folded and unfolded states are nearly equally adversely affected by macromolecular crowding. For any environmental variable, it is the balance of its effects on the folded and unfolded states that determines the outcome on the folding stability.

Ficoll↗
PubMed2006

Activity of Subtilisin Carlsberg in macromolecular crowding.

Enzymatic activity of a proteolytic enzyme Subtilisin Carlsberg (SC) in anionic sodium dodecyl sulfate (SDS) micellar medium has been explored and found to be retarded compared to that in bulk buffer. Circular dichroism (CD) study reveals that SDS, which is a potential protein denaturant, has an insignificant denaturation effect on SC. The structural integrity of the protein offers an opportunity to study the functionality of the enzyme SC in a macromolecular crowding of micelles. Dynamic light scattering (DLS) data indicates no sandwich-like micelle-SC complex formation ruling out the possibility of interaction of the enzyme with the hydrophobic core of the micelle. However, steady state and time resolved emission studies on specific and nonspecific fluorescent probes indicate the proximity effect at the surface of the enzyme due to macromolecular crowding of the micelles. The agreement of retarded enzymatic activity in the micellar crowd with a theoretical model ascribed to the facts that substrates are compartmentalized in the micelles and enzyme interacts with the micelle through stern layer.

Circular Dichroism↗
PubMed2005

Swelling of hepatocytes injured by oxidative stress suggests pathological changes related to macromolecular crowding.

A fundamental property of living matter is the ability to establish and maintain order. Mild changes in cell volume have a role in metabolic control. Furthermore, cellular swelling is a way of living cells to react to a variety of stressors. Data from experimental pathology, biochemistry and biophysics and theoretical arguments from biology, biochemical evolution, cytology and biophysics are considered to attempt an integration of several current concepts on different subjects (intracellular compartmentation, cellular swelling, macromolecular crowding, perturbing and non-perturbing solutes). The purpose is to provide a framework for conceptualizing in modern terms the question whether cellular swelling induced by oxidative stress should be considered merely a cell adaptation balanced by antioxidant defenses and by other biochemical devices apt to preserve the intracellular environment and normal cell functioning, or whether swelling of high amplitude should be regarded as a true pathological change. The basic question dated 1982: "how crowded is the cytoplasm?" is a matter for discussion as far as swollen cells are concerned. This paper examines the liver for cellular swelling of high amplitude (about+30%) caused by iron or by thyroid hormone+iron (histological picture of "cloudy swelling") or the steatogenic poison CCl(4), also known as a source of oxyradicals, which causes an even more pronounced cellular swelling. In CCl(4)-toxic fatty liver the strong increase of tissue water is substantially masked by the parallel increase of tissue dry solids due to fat accumulation. This example of a "tissue dilution artefact" is discussed in connection with the increase of tissue water also in toxic fatty liver induced by white phosphorus and ethanol. In CCl(4)-toxic fatty liver the normal K(+)/Na(+) ratio (about 3) is substantially maintained, whereas the concentrations of the two cations ("perturbing osmolytes") in tissue water are noticeably decreased, a finding which was not further studied at the time the observations were made because biochemistry was not yet advanced enough to allow an explanation. Today, a logic hypothesis is that an increase of non-perturbing solutes such as taurine and betaine, maintains the physiological intracellular osmotic pressure and that the harmful effects of CCl(4) are limited because of the protective effects of these molecules and of molecular chaperones against damage by oxyradicals. However, as a consequence of cellular swelling, intracellular changes in ionic strength and macromolecular crowding should occur thus affecting enzyme activities. Models and techniques apt to investigate this problem experimentally are suggested.

Animals↗
PubMed2007

Shift in spatial scale in identifying crowded letters.

Crowding refers to the increased difficulty in identifying a letter flanked by other letters. The purpose of this study was to determine if the peak sensitivity of the human visual system shifts to a different spatial frequency when identifying crowded letters, compared with single letters. We measured contrast thresholds for identifying the middle target letters in trigrams, for a range of spatial frequencies, letter separations and letter sizes, at the fovea and 5 degrees eccentricity. Plots of contrast sensitivity vs. letter frequency exhibit spatial tuning, for all letter sizes and letter separations tested. The peak tuning frequency grows as the 0.6-0.7 power of the letter size, independent of letter separation. At the smallest letter separation, peak tuning frequency occurs at a frequency that is 0.17 octaves higher for flanked than for unflanked letters at the fovea, and 0.19 octaves at 5 degrees eccentricity. This finding suggests that the human visual system shifts its sensitivity toward a higher spatial-frequency channel when identifying letters in the presence of nearby letters. However, the size of the shift is insufficient to account for the large effect of crowding in the periphery.

Contrast Sensitivity↗
PubMed1995

Calculation of resonance energy transfer in crowded biological membranes.

Analytical and numerical models were developed to describe fluorescence resonance energy transfer (RET) in crowded biological membranes. It was assumed that fluorescent donors were linked to membrane proteins and that acceptors were linked to membrane lipids. No restrictions were placed on the location of the donor within the protein or the partitioning of acceptors between the two leaflets of the bilayer; however, acceptors were excluded from the area occupied by proteins. Analytical equations were derived that give the average quantum yield of a donor at low protein concentrations. Monte Carlo simulations were used to generate protein and lipid distributions that were linked numerically with RET equations to determine the average quantum yield and the distribution of donor fluorescence lifetimes at high protein concentrations, up to 50% area fraction. The Monte Carlo results show such crowding always reduces the quantum yield, probably because crowding increases acceptor concentrations near donor-bearing proteins; the magnitude of the reduction increases monotonically with protein concentration. The Monte Carlo results also show that the distribution of fluorescence lifetimes can differ markedly, even for systems possessing the same average lifetime. The dependence of energy transfer on acceptor concentration, protein radius, donor position within the protein, and the fraction of acceptors in each leaflet was also examined. The model and results are directly applicable to the analysis of RET data obtained from biological membranes; their application should result in a more complete and accurate determination of the structures of membrane components.

Biophysical Phenomena↗
PubMed2002

Accelerated alpha-synuclein fibrillation in crowded milieu.

Parkinson's disease is the second most common age-related neurodegenerative disease, resulting from loss of dopaminergic neurons in the substantia nigra. The aggregation and fibrillation of alpha-synuclein has been implicated as a causative factor in the disease, and the process of fibril formation has been intensively studied in vitro with dilute protein solutions. However, the intracellular environment of proteins is crowded with other macromolecules, whose concentration can reach 400 g/l. To address this discrepancy, the effect of molecular crowding on alpha-synuclein fibrillation has being studied. The addition of high concentrations of different polymers (proteins, polysaccharides and polyethylene glycols) dramatically accelerated alpha-synuclein fibrillation in vitro. The magnitude of the accelerating effect depended on the nature of the polymer, its length and concentration. Our results suggest that the major factor responsible for the accelerated fibrillation under crowded conditions is the excluded volume.

Chemical Phenomena↗
PubMed2003

Emergency department crowding: consensus development of potential measures.

STUDY OBJECTIVE: We identify measures of emergency department (ED) and hospital workflow that would be of value in understanding, monitoring, and managing crowding. METHODS: A national group of 74 experts developed 113 potential measures using a conceptual model of ED crowding that segmented the measures into input, throughput, and output categories. Ten investigators then used group consensus methods to revise and consolidate them into a refined set of 30 measures that were rated by all 74 experts, who used a magnitude estimation technique on a Web site. Each measure was compared with a standard to obtain numeric ratings for feasibility, affordability, early warning potential, long-term planning potential, a summary rating of operational usefulness, and research potential. After review of the comprehensiveness of the resulting measures, 8 additional measures were developed and also rated by all reviewers. RESULTS: The original set of 113 measures (46 input, 35 throughput, and 32 output) was reduced to 38 through the iterative revision and rating process (15 input, 9 throughput, and 14 output). Summary scores in each rating category showed significant variation in ratings among the various potential measures. For measures that address similar concepts, the priority ranking depended on the rating category chosen. CONCLUSION: The final 38 measures of ED and hospital workflow provide a useful pool from which EDs and policymakers can draw to improve their ability to understand and address the issue of ED crowding. These measures require rigorous testing for feasibility, reliability, and value.

Consensus↗
PubMed2003

Prospects of electron cryotomography to visualize macromolecular complexes inside cellular compartments: implications of crowding.

Electron cryotomography has unique potential for three-dimensional visualization of macromolecular complexes at work in their natural environment. This approach is based on reconstructing three-dimensional volumes from tilt series of electron micrographs of cells preserved in their native states by vitrification. Resolutions of 5-8 nm have already been achieved and the prospects for further improvement are good. Since many intracellular activities are conducted by complexes in the megadalton range with dimensions of 20-50 nm, current resolutions should suffice to identify many of them in tomograms. However, residual noise and the dense packing of cellular constituents hamper interpretation. Recently, tomographic data have been collected on vitrified eukaryotic cells (Medalia et al., Science (2002) in press). Their cytoplasm was found to be markedly less crowded than in the prokaryotes previously studied, in accord with differences in crowding between prokaryotic and eukaryotic cells documented by other (indirect) biophysical methods. The implications of this observation are twofold. First, complexes should be more easily identifiable in tomograms of eukaryotic cytoplasm. This applies both to recognizing known complexes and characterizing novel complexes. An example of the latter-a 5-fold symmetric particle is-given. Second, electron cryotomography offers an incisive probe to examine crowding in different cellular compartments.

Animals↗