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Cell membrane shape control--effects of chloromethyl ketone peptides.

The shape of the human erythrocyte is normally maintained in vivo as a biconcave disc for 120 days. In vitro, the cell shape can be altered readily by amphipathic compounds; however, given time and an energy source, the cells can recover the discoid morphology. An active shape control mechanism is postulated to regulate erythrocyte shape. The shape recovery process is a necessary element in reversing perturbations of shape and is basic to our understanding of how membrane shape is altered. We report here that the process of shape recovery from crenation is dramatically accelerated upon pretreatment of the cells with micromolar (20-100 microM) concentrations of chloromethyl ketone peptides [such as N-alpha-tosyl-L-phenylalanine-chloromethyl ketone (tos-pheCH2Cl)]. Such pretreatments do not appear to affect cellular viability, as judged by their normal biconcave disc shape, their sensitivity to crenators, their lactic acid production, or the ATP-dependent shape change of the purified membranes. Treatment with high concentrations of tos-pheCH2Cl does cause normal cells to become stomatocytic by an energy-requiring process, i.e., it requires glucose, incubation at 37 degrees C, and will not occur in ATP-depleted cells. We suggest that the chloromethyl ketone peptides affect a metabolic process that is associated with the hexose monophosphate (HMP) shunt. Through the alteration of the HMP shunt metabolism, they modify an active stomatocytic process in the erythrocyte that can correct for the perturbation caused by crenators. Implications of these findings for analogous phenomena in cultured cells are discussed.

2,4-Dinitrophenol↗

Lipid molecular shapes and membrane architecture.

A simple biomembrane like erythrocyte contains well over hundred lipid species with diverse molecular shapes differing in the number of acyl chains, chain length, unsaturation and head group composition. A delicate balance between these molecular shapes is necessary in order to have a functional membrane. It is well established that the activities of a number of membrane-bound enzymes and other properties such as aggregation, spontaneous vesiculation, pathophysiological properties and lipid-protein interactions of lipids depend on the acyl chain length, unsaturation and head group composition. In fact, the molecular shape of a phospholipid molecule, as modulated by changes in chain length, unsaturation and head group composition, is probably what is affecting the above mentioned properties. The molecular shape of a lipid depends on a dimensionless packing parameter, S, the value of which influences the size and shape of aggregate formed upon hydration. In fact, the additivity of S values of lipid mixtures explains a number of experimental observations. The molecular shape concept, although very simple, explains many membrane phenomena like complementarity of molecular shapes of non-bilayer lipids to form stable bilayers. Membrane permeability is controlled to a large extent by lipid packing which depends upon molecular shapes. In fact, membranes maintain their lamellar structure by delicately balancing the composition of bilayer-forming and non-bilayer-forming lipids indicating that complementarity of molecular shapes is essential to maintain the permeability barrier. Based on this, the complementary molecular shape model of cell membrane is proposed.

Animals↗

S-GMAS: Genome-Wide Mediation Analysis With Brain Subcortical Shape Mediators.

Mediation analysis is widely utilized in neuroscience to investigate the role of brain image phenotypes in the neurological pathways from genetic exposures to clinical outcomes. However, it is still difficult to conduct mediation analyses with whole genome-wide exposures and brain subcortical shape mediators due to several challenges including (i) large-scale genetic exposures, that is, millions of single-nucleotide polymorphisms (SNPs); (ii) nonlinear Hilbert space for shape mediators; and (iii) statistical inference on the direct and indirect effects. To tackle these challenges, this paper proposes a genome-wide mediation analysis framework with brain subcortical shape mediators. First, to address the issue caused by the high dimensionality in genetic exposures, a fast genome-wide association analysis is conducted to discover potential genetic variants with significant genetic effects on the clinical outcome. Second, the square-root velocity function representations are extracted from the brain subcortical shapes, which fall in an unconstrained linear Hilbert subspace. Third, to identify the underlying causal pathways from the detected SNPs to the clinical outcome implicitly through the shape mediators, we utilize a shape mediation analysis framework consisting of a shape-on-scalar model and a scalar-on-shape model. Furthermore, the bootstrap resampling approach is adopted to investigate both global and spatial significant mediation effects. Finally, our framework is applied to the corpus callosum shape data from the Alzheimer's Disease Neuroimaging Initiative.

Humans↗

Shape-engineered fibroblasts: cell elasticity and actin cytoskeletal features characterized by fluorescence and atomic force microscopy.

The regulation of cell shape, which determines cell behaviors including adhesion, spreading, migration, and proliferation in an engineered artificial extracellular milieu, is an important task in tissue engineering and in development of functional biomaterials. To deepen the understandings of shape-dependent cell mechanics, the cell elasticity and structural features of the actin cytoskeleton (CSK) were characterized for shape-engineered fibroblasts; round and spindle-shaped cells cultured on photolithographically microprocessed surfaces, employing the cellular microindentation tests and fluorescence observation of actin CSK by the combination of atomic force microscopy (AFM) and fluorescence microscopy (FM). The relationships among cell elasticity, the structural features of actin CSK, and engineered cell shape were analyzed and compared with those of control cells that had been cultured on nonprocessed surfaces (termed naturally extended cells). Results showed that the spindle-shaped cells with sparse or no apical stress fibers (ASFs) exhibited similar stiffness to that of the naturally extended cells with dense ASFs. The elasticity of spindle-shaped cells was affected only slightly by the stress fiber (SF) density, which is in marked contrast to the significant correlation shown between cell elasticity and SF density in naturally extended cells. This result implies that the elasticity of regionally restricted adhesion-surface-induced shape-engineered cells, particularly of highly elongated cells, is affected predominantly by cell shape rather than by structural features of SFs.

3T3 Cells↗

Shape memory of human red blood cells.

The human red cell can be deformed by external forces but returns to the biconcave resting shape after removal of the forces. If after such shape excursions the rim is always formed by the same part of the membrane, the cell is said to have a memory of its biconcave shape. If the rim can form anywhere on the membrane, the cell would have no shape memory. The shape memory was probed by an experiment called go-and-stop. Locations on the membrane were marked by spontaneously adhering latex spheres. Shape excursions were induced by shear flow. In virtually all red cells, a shape memory was found. After stop of flow and during the return of the latex spheres to the original location, the red cell shape was biconcave. The return occurred by a tank-tread motion of the membrane. The memory could not be eliminated by deforming the red cells in shear flow up to 4 h at room temperature as well as at 37 degrees C. It is suggested that 1). the characteristic time of stress relaxation is >80 min and 2). red cells in vivo also have a shape memory.

Biomechanical Phenomena↗

Histamine H4 receptor mediates eosinophil chemotaxis with cell shape change and adhesion molecule upregulation.

1. During mast cell degranulation, histamine is released in large quantities. Human eosinophils were found to express histamine H(4) but not H(3) receptors. The possible effects of histamine on eosinophils and the receptor mediating these effects were investigated in our studies. 2. Histamine (0.01-30 microm) induced a rapid and transient cell shape change in human eosinophils, but had no effects on neutrophils. The maximal shape change was at 0.3 microm histamine with EC(50) at 19 nm. After 60 min incubation with 1 microm histamine, eosinophils were desensitized and were refractory to shape change response upon histamine restimulation. Histamine (0.01-1 microm) also enhanced the eosinophil shape change induced by other chemokines. 3. Histamine-induced eosinophil shape change was mediated by the H(4) receptor. This effect was completely inhibited by H(4) receptor-specific antagonist JNJ 7777120 (IC(50) 0.3 microm) and H(3)/H(4) receptor antagonist thioperamide (IC(50) 1.4 microm), but not by selective H(1), H(2) or H(3) receptor antagonists. H(4) receptor agonists imetit (EC(50) 25 nm) and clobenpropit (EC(50) 72 nm) could mimic histamine effect in inducing eosinophil shape change. 4. Histamine (0.01-100 microm) induced upregulation of adhesion molecules CD11b/CD18 (Mac-1) and CD54 (ICAM-1) on eosinophils. This effect was mediated by the H(4) receptor and could be blocked by H(4) receptor antagonists JNJ 7777120 and thioperamide. 5. Histamine (0.01-10 microm) induced eosinophil chemotaxis with an EC(50) of 83 nm. This effect was mediated by the H(4) receptor and could be blocked by H(4) receptor antagonists JNJ 7777120 (IC(50) 86 nm) and thioperamide (IC(50) 519 nm). Histamine (0.5 microm) also enhanced the eosinophil shape change induced by other chemokines. 6. In conclusion, we have demonstrated a new mechanism of eosinophil recruitment driven by mast cells via the release of histamine. Using specific histamine receptor ligands, we have provided a definitive proof that the H(4) receptor mediates eosinophil chemotaxis, cell shape change and upregulation of adhesion molecules. The effect of H(4) receptor antagonists in blocking eosinophil infiltration could be valuable for the treatment of allergic diseases. The histamine-induced shape change and upregulation of adhesion molecules on eosinophils can serve as biomarkers for clinical studies of H(4) receptor antagonists.

Cell Adhesion Molecules↗

Diagnostic differences in mandibular P4 shape between Neandertals and anatomically modern humans.

This study uses elliptical Fourier analysis to quantify shape differences observed in the P(4) crown of Neandertals and anatomically modern humans. Previously, P(4) shape was assessed qualitatively, and results suggested marked differences between Neandertals and anatomically modern humans (Bailey [2002] New Anat. 269:148-156). The goal of this study was to investigate the P(4) shape in more detail, quantifying it in order to determine its utility for taxonomic classification and phylogenetic analysis. A comparison of mean shapes confirms that the mesiolingual portion of the P(4) is truncated in Neandertals, and that this produces a distinctively asymmetrical P(4). A randomization test confirms that the shape difference between Neandertals and anatomically modern humans is significant. Principal component and discriminant function analyses indicate that the relative size of the lingual portion of the tooth also affects tooth shape, with the lingual portion of the Neandertal P(4) being narrower than that of anatomically modern humans. Classification of P(4) crown shapes using discriminant functions analysis is far from perfect. While 86.4% of the teeth were correctly classified, classification was much better for anatomically modern humans (98.1%) than it was for Neandertals (65%). Fortunately, crown shape is but one of several diagnostic characters of the P(4) crown. P(4) crown asymmetry can be added to the growing list of dental morphological characters distinguishing Neandertals from anatomically modern humans. Moreover, based on a comparison of mean tooth shapes in fossil and recent humans, symmetry, rather than asymmetry, appears to be the primitive state, and the high frequency of P(4) asymmetry is likely derived in Neandertals.

Animals↗

Shape behavior of lipid vesicles as the basis of some cellular processes.

The basic principles that govern the shape behavior of phospholipid vesicle shapes are discussed. The important membrane parameters of the system are defined by presenting the expressions for the relevant contributions to the system's mechanical energy. In the description of the rather unique shape behavior of lipid vesicles, the emphasis is on providing a qualitative understanding of the dependence of vesicle shape on the parameters of the system. The vesicle shape behavior is then related to biologically important phenomena. Some examples are given of how the results of the shape behavior of lipid vesicles can be applied to the analysis of cellular systems. Red blood cell shape and shape transformations, vesicle fission and fusion processes, and the phenomenon of cellular polarity are considered. It is reasoned that the current biological processes that involve changes of membrane conformation may have their origin in the general shape behavior of closed lamellar membranes.

Algorithms↗

Shape diversity among chick retina Müller cells and their postnatal differentiation.

It is currently believed that in each vertebrate species Müller cells in the central retina constitutes a fairly homogeneous population from the morphologic point of view and that particularly the chick Müller cell attains full shape differentiation at prenatal stages. However, in this study of the chick retina, from day 1 to day 55 of life, we show that there is a large variety of Müller cell shapes and that many of them complete shape differentiation postnatally. We used a cell dissociation method that preserves the whole shape of the Müller cells. Unstained living and unstained fixed cells were studied by phase-contrast microscopy, and fixed cells immunostained for intermediate filaments of the cytoskeleton were studied by fluorescence microscopy. Our results show that (1) Müller cell shapes vary in the origination of the hair of vitread processes, in the shape of the ventricular (outer or apical) process, in the presence or absence of an accessory process, as well as in the number and shape of processes leaving from the ventricular process at the level of the outer nuclear and outer plexiform layers (ONL/OPL); (2) during the first month of life, many Müller cells differentiate the portion of the ventricular process that traverses the ONL, most Müller cells differentiate the ONL/OPL processes, and all Müller cells differentiate the thin short lateral processes leaving from the vitread hair processes at the level of the inner plexiform layer (IPL). The number of cells differing in the shape of the ventricular process and that of cells with and without accessory process were estimated. The spatial relationship between the outer portion of the ventricular process of the Müller cell and the photoreceptor cells was also studied. Our results show that the branching of the ventricular process and the refinement of Müller cell shape is achieved without apparent participation of growth cones. We give a schematic view of how the branching of the ventricular process might take place and propose the size increase of photoreceptor soma as a factor responsible for this branching.

Aging↗

Regulation of receptor-mediated shape change in astroglial cells.

Activation of adenylate cyclase in astroglial cells in culture results in a rapid change in cell shape that appears to occur by the active movement of cytoplasm from peripheral cell regions to the perinuclear space with processes being formed along regions that remain extended. Three series of experiments were designed to determine how shape change occurred. First, the Ca(2+)-dependency of shape change was determined by reducing intracellular Ca2+ concentrations to less than or equal to 50 nM or increasing intracellular Ca2+ concentrations to greater than or equal to 1 microM. Neither of these changes significantly affected the rate of receptor-mediated shape change. Second the role that longer-lived, acetylated microtubules play in receptor-mediated shape change was assessed by visualizing microtubules using a polyclonal antibody to brain 6S tubulin or a monoclonal antibody to oligomers of tubulin to monitor total tubulin distribution and a monoclonal antibody to acetylated tubulin to describe the distribution of these microtubules. Three-dimensional distribution of microtubules was observed by optical sectioning of cultures using a laser scanning confocal imaging system. The distribution of acetylated tubules in control cells was similar to that observed with the antibodies to tubulin. Following treatment with 100 nM isoproterenol to stimulate shape change, there was a dramatic redistribution of microtubules; however, the distribution of acetylated tubules was again similar to the total microtubules. Analysis of the optical sections recorded using the confocal attachment revealed that while control cells were relatively flat (cell height = 4 microns), the perinuclear region of isoproterenol-treated cells extended much higher above the substrate (cell height = 13 microns). Third, the role of microtubule assembly and disassembly were assessed using colchicine and taxol. Results from these experiments suggest that microtubule reassembly is necessary for receptor-mediated shape change. Control experiments indicated that colchicine or taxol treatment did not inhibit either cAMP synthesis or another cAMP-dependent process, receptor-mediated taurine release. Together these results indicate that receptor-mediated shape change in astroglial cells occurs by a Ca(2+)-independent mechanism that results in active movement of cytoplasm to the perinuclear region. This process is dependent on microtubule reassembly suggesting that shape change may occur by active movement of material along microtubules or by microtubule redistribution.

Acetylation↗

Prostaglandins change cell shape and increase intercellular gap junctions in osteoblasts cultured from rat fetal calvaria.

Mounting experimental evidence indicates that osteoblasts may be cellular intermediaries in the local activation of bone remodeling. To elucidate the role of these cells in activation, we examined the effects of prostaglandins (PGs), known resorption stimulators, on cell shape and intercellular junctional relationships in osteoblasts cultured from rat fetal calvaria. Exposure to PGE2 and PGE1, promoters of bone resorption, rapidly (within 20 min) converted the osteoblasts from a flattened to a stellate shape (shape change), and markedly increased the appearance of intercellular (gap) junctions within 10 min. Both effects were directly related to the prostaglandin concentration, as little as 1 nM being effective. PGE1, but not PGB1, PGF1 alpha, PGD2, and PGF2 alpha, mimicked the substantial effect of PGE2 on shape change. Shape change and gap junction formation appear to arise independently. PTH, an inducer of shape change, did not affect the number of gap junctions appreciably. Colchicine, a microtubule polymerization inhibitor, and trifluoperazine, an inhibitor of calmodulin action, blunted PGE2-mediated shape change but not the effect of PGE2 on gap junctions. Shape change and gap junction formation may be important events in local activation, shape changes in surface osteoblasts serving to expose bone surfaces which are chemotactic for osteoclasts and gap junctions propagating locally initiated activation messages.

Animals↗

Relationship between pH, sodium, and shape changes in chemotactic-factor-stimulated human neutrophils.

The relationship between the chemotactic-factor-elicited changes in the intracellular pH and the shape of human neutrophils was investigated using simultaneous measurements of both parameters. The results demonstrate first that fMet-Leu-Phe and leukotriene B4 elicit qualitatively similar pH and shape change responses from the neutrophils. A relationship between the chemoattractant-elicited decrease in cytoplasmic pH and the shape changes is indicated by several findings including: 1) the similarities in the time courses of the two responses, 2) the ability of propionic acid to induce a transient and pertussis-toxin-sensitive shape change response, and 3) the ability of the calcium ionophore A23187 to similarly induce both responses under conditions when the degranulation is minimized. On the other hand, several other results indicate that the drop in pH is not a sufficient condition for the chemotactic-factor-stimulated shape changes. These include: 1) the ability of pertussis toxin to inhibit the shape changes induced by propionic acid and by A23187 without affecting the drop in pH, and 2) the observation that the drop in pH induced by propionic acid persists significantly longer than the shape change. Increasing the cytoplasmic pH by adding ammonium chloride was also found to cause shape changes in the neutrophils. The response to the base differs in two important aspects from that caused by propionic acid: it is pertussis-toxin-insensitive, and it is long-lived. Chemotactic factors have been found to induce a shape change under conditions when the internal pH was artificially increased or decreased, indicating that it is not the absolute cytoplasmic pH that represents the internal signalling parameter. The results are discussed in terms of the activation of the cytoskeletal network of the neutrophils by chemotactic factors.

Ammonium Chloride↗

Quantitative two-dimensional echocardiographic assessment of left ventricular shape in ischemic heart disease.

Since data on systematic noninvasive quantitative assessment of left ventricular (LV) shape in ischemic heart disease are very scanty, we used new size-independent LV shape descriptors to study the correlation between LV shape and LV ejection fraction in 46 men with a 70% or more stenosis of one or more coronary arteries on angiography. Angiocardiographic LV ejection fraction was over 55% [mean: 69 (+/- 6 SD)%] in 20 patients (group I), and under 55% [mean: 43 (+/- 9 SD)%] in 26 patients (group II). Our LV shape descriptors showed statistically significant differences between group I and group II patients, indicating a more spherical LV shape in group II (p values less than 0.005). Within the whole patient group (N = 46), and also within the subgroup of patients with regional LV hypokinesis or akinesis (N = 26), correlation between LV shape alteration (toward the spherical) and LV ejection fraction was not strong; it was better in the subgroup (N = 10) with regional LV wall dyskinesis. There was poor correlation between angiocardiographic LV chamber volume and alteration (toward spherical) in LV shape. Thus, we have documented quantitatively that patients with ischemic heart disease and low LV ejection fraction tend to have a more spherical LV shape than patients with a normal ejection fraction. Quantification of LV shape based on simple, easily made two-dimensional echocardiographic measurements may help in noninvasive identification of patients with deteriorating LV function, especially when a change toward the spherical is detected in serial echocardiograms.

Adult↗

Shaping of gelling biopolymer drops in an elongation flow.

Shaping, defined as deformation in combination with gel formation of gelatine and kappa-carrageenan drops in an elongation flow, was studied. The focus was to investigate the possibility of shaping and fixating small drops in the diameter range 20 to 229 mum. In the shaping progress and the influence of experimental properties, the viscosity, temperature, and flow of the deforming fluid were examined on the final drop shape. In the experiments a hot emulsion of an aqueous biopolymer solution in silicone oil was injected into cold silicone oil where a deforming elongation flow field existed. After injection, a temperature decrease in the drops resulted in a gel formation of the biopolymer and a fixation of the deformed drop in the flow. The shape was measured and the effect on the drop aspect ratio was determined by image analysis. Over the total drop diameter range, kappa-carrageenan was more ellipsoid-shaped than gelatine, with a maximum aspect ratio of 6 compared to 4 for gelatine. For small drops, around 22 mum, it is possible to shape kappa-carrageenan, but for gelatine small drops tend to be unaffected. An increase in viscosity, temperature, and flow resulted in an increase in the final fixated shape of the drops. The differences in drop deformation between the biopolymers were explained by drop-viscosity/oil differences and differences in the kinetics of gel formation. The different gel formation kinetics resulted in a short, well-defined, shaping process for kappa-carrageenan, while for gelatine the process was more complex, with both deformation and relaxation present at different stages.

Biopolymers↗

Coupled multi-shape model and mutual information for medical image segmentation.

This paper presents extensions which improve the performance of the shape-based deformable active contour model presented earlier in [9]. In contrast to that work, the segmentation framework that we present in this paper allows multiple shapes to be segmented simultaneously in a seamless fashion. To achieve this, multiple signed distance functions are employed as the implicit representations of the multiple shape classes within the image. A parametric model for this new representation is derived by applying principal component analysis to the collection of these multiple signed distance functions. By deriving a parametric model in this manner, we obtain a coupling between the multiple shapes within the image and hence effectively capture the co-variations among the different shapes. The parameters of the multi-shape model are then calculated to minimize a single mutual information-based cost functional for image segmentation. The use of a single cost criterion further enhances the coupling between the multiple shapes as the deformation of any given shape depends, at all times, upon every other shape, regardless of their proximity. We demonstrate the utility of this algorithm to the segmentation of the prostate gland, the rectum, and the internal obturator muscles for MR-guided prostate brachytherapy.

Algorithms↗

Shape discrimination in the hippocampus using an MDL model.

We extend recent work on building 3D statistical shape models, automatically, from sets of training shapes and describe an application in shape analysis. Using an existing measure of model quality, based on a minimum description length criterion, and an existing method of surface re-parameterisation, we introduce a new approach to model optimisation that is scalable, more accurate, and involves fewer parameters than previous methods. We use the new approach to build a model of the right hippocampus, using a training set of 82 shapes, manually segmented from 3D MR images of the brain. We compare the results with those obtained using another previously published method for building 3D models, and show that our approach results in a model that is significantly more specific, general, and compact. The two models are used to investigate the hypothesis that there are differences in hippocampal shape between age-matched schizophrenic and normal control subgroups within the training set. Linear discriminant analysis is used to find the combination of shape parameters that best separates the two subgroups. We perform an unbiased test that shows there is a statistically significant shape difference using either shape model, but that the difference is more significant using the model built using our approach. We show also that the difference between the two subgroups can be visualised as a mode of shape variation.

Algorithms↗

A radiographic analysis of the relationship between the size and shape of the intercondylar notch and anterior cruciate ligament injury.

Notch-view radiographs were obtained from 108 persons with anterior cruciate ligament (ACL) injuries (55 women, 53 men) and 186 with intact ACL (94 women, 92 men). Notch width, femur width, and notch width index were determined from each of the 294 radiographs. The notch was also categorized as either A-shaped or non-A-shaped. Intrarater and interrater reliability ranged from 0.82 to 0.99 for notch width and femur width, respectively. Reliability within and between raters for the classification of notch shape ranged from 0.80 to 1.0. Notch width was significantly influenced by a 10 degree change in knee angle when repeated radiographs were taken. Femur width was not affected by knee angle across this range. Analysis revealed a higher proportion of A-shaped notches among women than men. However, notch shape was not related to injury status. Results showed a smaller notch width and notch width index in ACL-injured patients regardless of notch shape or gender. A-shaped notches were smaller than non-A-shaped notches regardless of injury status or gender. Both notch width and notch width index were found to be significant indicators of ACL injury. Knowledge of the shape of the notch added no useful information in differentiating patients based on injury status. Thus, regardless of gender, individuals who possess smaller notch dimensions appear to be at greater risk of injury than individuals with larger notches.

Adolescent↗

Distortions in the visual perception of shape.

It is known that visual illusions lead to a distorted perception of the length and orientation of lines, but it is not clear how these illusions affect the appreciation of the shape of closed forms. In this study two experiments were performed to characterize distortions in the visual perception of the shape of quadrilaterals and the extent to which these distortions were similar to the distortions of haptically sensed shapes. In the first experiment human subjects were presented with two quadrilaterals side by side on a computer monitor. One was a reference shape; the other was rotated and distorted relative to the first. The subjects used the computer mouse to adjust the corners of the distorted quadrilateral to match the shape of the target quadrilateral. They made consistent errors on this task: the adjusted quadrilateral was about 2% wider and about 2% shorter than the veridical shape. Furthermore, subjects adjusted the inner angles of the quadrilateral to make them closer to 90 degrees . The first type of error was also present in a second experiment in which, in a two-alternative forced-choice paradigm, subjects viewed a reference shape and were asked to indicate which of two transiently presented quadrilaterals was closest to the target shape. The width/height errors and the inner angle errors were comparable to those described previously when subjects felt the outline of a quadrilateral and then drew its reproduction in the absence of vision, suggesting that the distortion occurs in the process of remembering the shape.

Adult↗