The Emotional Eye: Red Sclera as a Uniquely Human Cue of Emotion

The sclera, the eye's tough white outer layer, provides the ground necessary for the display of its own color and that of the overlying membrane, the conjunctiva. This study evaluated the sclera as a cue of emotion by contrasting the ratings of 38 subjects for the level of anger, fear, sadness, disgust, happiness, or surprise of normal (untinted) eye images with copies of those images that were reddened by digital editing. Subjects rated individuals with reddened sclera as having more anger, fear, disgust, and sadness, and less happiness than those with normal, untinted sclera. Surprise was the only emotion unaffected by scleral redness. Humans, but not other primates, have evolved the white sclera necessary to display the blood flow in the overlying conjunctiva that produces the redness associated with certain emotional states.     

Red, Yellow, and Super-White Sclera

The sclera, the eye’s tough outer layer, is, among primates, white only in humans, providing the ground necessary for the display of colors that vary in health and disease. The current study evaluates scleral color as a cue of socially significant information about health, attractiveness, and age by contrasting the perception of eyes with normal whites with copies of those eyes whose whites were reddened, yellowed, or further whitened by digital editing. Individuals with red and yellow sclera were rated to be less healthy, less attractive, and older than individuals with untinted control sclera. Individuals with whitened, “super-white” sclera were rated as younger, although not more healthy or attractive, than controls. In humans, clear, white sclera may join such traits as smooth skin and long, lustrous hair as signs of health, beauty, and reproductive fitness. The evolution of a white sclera may have contributed to the emergence of humans as a social species.     

The Expression of Tenascin-X in Developing and Adult Rat and Human Eye

Tenascin-X has been studied in developing and adult rat eye and in foetal and adult human eyes, using immunohistochemistry and frozen sections. The data were compared with the distribution of tenascin-C. The immunoreactivity for tenascin-X was seen in a basement membrane-like feature in different structures of embryonic (E) day 16–17 rat eyes. Postnatal (P) day 2 and older rat eyes showed immunoreactivity for tenascin-X in different connective tissues. In the epithelial basement membrane zone of the cornea, immunostaining was positive in P5 eyes, negative in P10 and P15 eyes and again positive in P30 and adult eyes. In the 20-week-old human foetus, immunoreactivity for the tenascin was seen in the posterior parts of the conjunctival stroma adjacent to the sclera and in a basement membrane-like fashion in anterior conjunctiva. In the adult human eye, immunoreactivity for tenascin-X was seen in the anterior one-third stroma of cornea as thin fibrils, in the stroma of the limbus and conjunctiva, and in blood vessels. Immunostaining for tenascin-C was seen in the posterior aspect of the further cornea, and in mesenchyme adjacent to cornea in E16–17 rat eyes. Corneal keratocytes and Descemet's membrane showed immunoreactivity for tenascin-C in P2–P15 rat eyes. Sclera and the junction of the cornea, and sclera expressed tenascin-C in P2 and older rat eyes. In human foetal eyes, immunostaining for tenascin-C was seen in the anterior parts of the corneal stroma, in the basement membrane zone and Bowman's membrane of the corneal epithelium, in the posterior one-fifth of the corneal stroma and the sclera starting from the junction of the cornea and sclera. In normal human adult eyes, immunostaining for tenascin-X was seen in the anterior one-third stroma of cornea, in the stroma of limbus and conjunctiva, and in blood vessels. The association of tenascin-X and basement membranes in early development evokes a question of its potential function in the development of the basement membrane. The results also suggest the association of tenascin-X with connective tissue development as well as the association of tenascin-C with the migration of keratocytes during the development of the corneal stroma.     

Reliance on head versus eyes in the gaze following of great apes and human infants: the cooperative eye hypothesis

As compared with other primates, humans have especially visible eyes (e.g., white sclera). One hypothesis is that this feature of human eyes evolved to make it easier for conspecifics to follow an individual's gaze direction in close-range joint attentional and communicative interactions, which would seem to imply especially cooperative (mututalistic) conspecifics. In the current study, we tested one aspect of this cooperative eye hypothesis by comparing the gaze following behavior of great apes to that of human infants. A human experimenter "looked" to the ceiling either with his eyes only, head only (eyes closed), both head and eyes, or neither. Great apes followed gaze to the ceiling based mainly on the human's head direction (although eye direction played some role as well). In contrast, human infants relied almost exclusively on eye direction in these same situations. These results demonstrate that humans are especially reliant on eyes in gaze following situations, and thus, suggest that eyes evolved a new social function in human evolution, most likely to support cooperative (mututalistic) social interactions.     

Unique morphology of the human eye and its adaptive meaning: comparative studies on external morphology of the primate eye

In order to clarify the morphological uniqueness of the human eye and to obtain cues to understanding its adaptive significance, we compared the external morphology of the primate eye by measuring nearly half of all extant primate species. The results clearly showed exceptional features of the human eye: (1) the exposed white sclera is void of any pigmentation, (2) humans possess the largest ratio of exposed sclera in the eye outline, and (3) the eye outline is extraordinarily elongated in the horizontal direction. The close correlation of the parameters reflecting (2) and (3) with habitat type or body size of the species examined suggested that these two features are adaptations for extending the visual field by eyeball movement, especially in the horizontal direction. Comparison of eye coloration and facial coloration around the eye suggested that the dark coloration of exposed sclera of nonhuman primates is an adaptation to camouflage the gaze direction against other individuals and/or predators, and that the white sclera of the human eye is an adaptation to enhance the gaze signal. The uniqueness of human eye morphology among primates illustrates the remarkable difference between human and other primates in the ability to communicate using gaze signals.     

Imaging depth extension of optical coherence tomography in rabbit eyes using optical clearing agents

Optical coherence tomography has become an indispensable diagnostic tool in ophthalmology for imaging the retina and the anterior segment of the eye. However, the imaging depth of optical coherence tomography is limited by light attenuation in tissues due to optical scattering and absorption. In this study of rabbit eye both ex vivo and in vivo, optical coherence tomography imaging depth of the anterior and posterior segments of the eye was extended by using optical clearing agents to reduce multiple scattering. The sclera, the iris, and the ciliary body were clearly visualized by direct application of glycerol at an incision on the conjunctiva, and the posterior boundary of sclera and even the deeper tissues were detected by submerging the posterior segment of eye in glycerol solution ex vivo or by retro-bulbar injection of glycerol in vivo. The ex vivo rabbit eyes recovered to their original state in 60 s after saline-wash treatment, and normal optical coherence tomography images of the posterior segment of the sample eyes proved the self-recovery of in vivo performance. Signal intensities of optical coherence tomography images obtained before and after glycerol treatment were compared to analysis of the effect of optical clearing. To the best of our knowledge, this is the first study for imaging depth extension of optical coherence tomography in both the anterior and posterior segments of eye by using optical clearing agents.     

表皮生长因子受体在正常大鼠眼组织中的表达

采用免疫组织化学方法观察了表皮生长因子受体在正常大鼠眼组织的表达。结果发现：角膜上皮的深层细胞和角膜缘上皮细胞、部分结膜上皮细胞和结膜下结缔组织内成纤维细胞均强烈表达表皮生长因子受体。结果显示：表皮生长因子受体阳性细胞主要分布于眼表层组织。这些细胞不仅是眼组织损伤后修复、而且是多种手术能否成功和某些疾病形成中起重要作用的细胞     

What is unique about the human eye? Comparative image analysis on the external eye morphology of human and nonhuman great apes

The gaze-signaling hypothesis and the related cooperative-eye hypothesis posit that humans have evolved special external eye morphology, including exposed white sclera (the white of the eye), to enhance the visibility of eye-gaze direction and thereby facilitate conspecific communication through joint-attentional interaction and ostensive communication. However, recent quantitative studies questioned these hypotheses based on new findings that certain features of human eyes are not necessarily unique among great ape species. Accordingly, there is currently a heated debate over whether external eye features of humans are distinct from those of other apes and how such distinguishable features contribute to the visibility of eye-gaze direction. The present study leveraged updated image analysis techniques to test the uniqueness of human eye features in facial images of great apes. Although many eye features were similar between humans and other great apes, a key difference was that humans have uniformly white sclera which creates clear visibility of both the eye outline and iris—the two essential features contributing to the visibility of eye-gaze direction. We then tested the robustness of the visibility of these features against visual noise, such as shading and distancing, and found that both eye features remain detectable in the human eye, while eye outline becomes barely detectable in other species under these visually challenging conditions. Overall, we identified that humans have unique external eye morphology among other great apes, which ensures the robustness of eye-gaze signals in various visual conditions. Our results support and also critically update the central premises of the gaze-signaling hypothesis.     

Material properties and effect of preconditioning of human sclera,optic nerve,and optic nerve sheath

The optic nerve (ON) is a recently recognized tractional load on the eye during larger horizontal eye rotations. In order to understand the mechanical behavior of the eye during adduction, it is necessary to characterize material properties of the sclera, ON, and in particular its sheath. We performed tensile loading of specimens taken from fresh postmortem human eyes to characterize the range of variation in their biomechanical properties and determine the effect of preconditioning. We fitted reduced polynomial hyperelastic models to represent the nonlinear tensile behavior of the anterior, equatorial, posterior, and peripapillary sclera, as well as the ON and its sheath. For comparison, we analyzed tangent moduli in low and high strain regions to represent stiffness. Scleral stiffness generally decreased from anterior to posterior ocular regions. The ON had the lowest tangent modulus, but was surrounded by a much stiffer sheath. The low-strain hyperelastic behaviors of adjacent anatomical regions of the ON, ON sheath, and posterior sclera were similar as appropriate to avoid discontinuities at their boundaries. Regional stiffnesses within individual eyes were moderately correlated, implying that mechanical properties in one region of an eye do not reliably reflect properties of another region of that eye, and that potentially pathological combinations could occur in an eye if regional properties are discrepant. Preconditioning modestly stiffened ocular tissues, except peripapillary sclera that softened. The nonlinear mechanical behavior of posterior ocular tissues permits their stresses to match closely at low strains, although progressively increasing strain causes particularly great stress in the peripapillary region.

     

Delivery of Topically Applied Calpain Inhibitory Peptide to the Posterior Segment of the Rat Eye

We developed an inhibitory peptide that specifically acts against mitochondrial μ-calpain (Tat-μCL, 23 amino acid, 2857.37 Da) and protects photoreceptors in retinal dystrophic rats. In the present study, we topically administered Tat-μCL to the eyes of Sprague-Dawley rats for 7 days to determine both the delivery route of the peptide to the posterior segment of the eye and the kinetics after topical application in adult rats. Distribution of the peptide was determined by immunohistochemical analysis, and enzyme-linked immune-absorbent assay was used to quantify the accumulation in the retina. Peptides were prominently detected in both the anterior and posterior segments of the eye at 1 h after the final eye drop application. Immunohistochemically positive reactions were observed in the retina, optic nerve, choroid, sclera and the retrobulbar tissues, even in the posterior portion of the eye. Immunoactivities gradually diminished at 3 and 6 h after the final eye drop. Quantitative estimations of the amount of peptide in the retina were 15.3, 5.8 and 1.0 pg/μg protein at 1, 3 and 6 h after the final instillation, respectively. Current results suggest that while the topically applied Tat-μCL peptide reaches the posterior segment of the retina and the optic nerve, the sufficient concentration (> IC50) is maintained for at least 6 h in the rat retina. Our findings suggest that delivery of topically applied peptide to the posterior segment and optic nerve occurs through the conjunctiva, periocular connective tissue, sclera and optic nerve sheath.     

The electroretinogram in multiple sclerosis and demyelinating optic neuritis

The electroretinogram (ERG) to flashes of white light presented under photopic conditions and the pattern reversal visual evoked potentials (PR-VEPs) from both eyes were recorded from 14 patients with multiple sclerosis (MS) with monocular demyelinating optic neuritis (DON) and from 11 patients soon after presenting with monocular demyelinating optic neuritis alone. Fifteen and 10 normal subjects, matched for age and sex, were used as controls for each group of patients respectively. In the DON group of patients and controls the flicker following ERG (FF-ERG) to white flashes of light at 40 Hz was also recorded. Skin electrodes and averaging procedures were used for all the recordings. The PR-VEP elicited with stimulation of the affected eye was absent or abnormally delayed, and the amplitude of the ‘b’ wave of ERG of the affected eye was diminished in all patients. The ‘b’ wave latency, however, was similar in both affected and non-affected eyes and the controls. There was no difference in ‘a’ wave amplitude and latency between eyes of patients and normal subjects. The FF-ERG in 8 out of 10 patients with satisfactory recordings was diminished in the affected eye. These results provide neurophysiological evidence that retinal damage is not due to loss of myelin but is an early feature of demyelinating optic neuritis. This damage preferentially affects the retinal elements associated with the generation of the ‘b’ wave of the ERG, probably the glial cells of Müller.     

Dynamic material properties of the human sclera

As a result of trauma, approximately 30,000 people become blind in one eye every year in the United States. A common injury prediction tool is computational modeling, which requires accurate material properties to produce reliable results. Therefore, the purpose of this study was to determine the dynamic material properties of the human sclera. A high-rate pressurization system was used to create dynamic pressure to the point of rupture in 12 human eyes. Measurements were obtained for the internal pressure, the diameter of the globe, the thickness of the sclera, and the changing coordinates of the optical markers using high-rate video. A relationship between true stress and true strain was determined for the sclera tissue in two directions. It was found that the average maximum true stress was 13.89±4.81 MPa for both the equatorial and meridional directions, the average maximum true strain along the equator was 0.041±0.014, and the average maximum true strain along the meridian was 0.058±0.018. Results show a significant difference in the maximum strain in the equatorial and meridional directions (p=0.02). In comparing these data with previous studies, it is concluded that the human sclera is both anisotropic and viscoelastic. The dynamic material properties presented in this study can be used for advanced models of the human eye to help prevent eye injuries in the future.     

超连续谱激光可见谱段致人眼眩目效应研究

本文采用超连续谱激光光源滤除其红外部分仅输出可见谱段部分,在不超过国家安全标准允许的最大辐照量条件下,以正入射方式照射人眼后,记录并分析在明、暗适应条件下中心极限视力恢复时间、中心近极限视力恢复时间和视觉后像持续时间,明确超连续谱激光可见谱段对人眼的眩目效果。明适应下激光照射0.1 s导致人眼中心极限视力恢复时间为31~119 s,中心近极限视力恢复时间为19~76 s;暗适应下激光照射0.1 s导致人眼中心极限视力恢复时间为26~223 s,中心近极限视力恢复时间为13~123 s;明、暗适应下导致人眼眩目效应的最小功率密度值分别为0.055 mW/cm^2和0.005 mW/cm^2。结果表明,超连续谱激光可见谱段对人眼有良好的眩目效果,可导致数十秒至数百秒的中心视力下降,随着照射功率密度增高,眩目效应增强,显示出较好的量效关系,且相同功率密度时暗适应下人眼的眩目效果优于明适应。该研究探究了明、暗适应条件下超连续谱激光对人眼眩目效应,明确了超连续谱激光与人眼眩目的量效关系。     

Apoptotic factors (Bcl-2 and Bax) and diabetic retinopathy in type 2 diabetes

The expression of apoptotic factors Bcl-2 and Bax were studied in the conjunctiva of diabetic patients with and without retinopathy. All patients underwent a complete ophthalmic examination including ocular fundus and retinal fluorescein angiography. The indirect immunoperoxidase method was performed on 15 normal conjunctiva taken during cataract surgery (group 1), on 40 eyes of 40 patients with type 2 diabetes without diabetic retinopathy (group 2) and 13 eyes of 13 patients with diabetic retinopathy (group 3). In normal human conjunctiva, Bax showed positive expression in epithelial, vascular and stromal cells whereas Bcl-2 staining was negative. In the conjunctiva of diabetic patients without diabetic retinopathy, Bax was widely, and strongly, expressed in epithelial cells, vascular endothelial cells, fibroblasts and infiltrating cells such as macrophages. For patients with diabetic retinopathy, Bax was consistently strong to very strong. Bcl-2 protein expression became weak to negative for diabetic patients both with and without diabetic retinopathy. Immunoreactivity was not correlated between Bcl-2 and Bax in the conjunctiva of diabetic patients. Bax was always localized in tissues characterized by a high rate of apoptosis, whereas, Bcl-2 was absent. Our results suggest that diabetic human conjunctiva, with its inflammatory phenomena, is considered as a privileged target for programmed cell death.     

Increased aggrecan (cartilage proteoglycan) production in the sclera of myopic chicks

A previously characterized chick model of myopia was used to evaluate biochemical changes in the sclera which are associated with ocular enlargement and myopia. Chicks were monocularly occluded for 10 days and the DNA, hydroxyproline, and glycosaminoglycan contents of the sclera were compared between the normal and the myopic eyes. No significant differences could be detected in total DNA or hydroxyproline content. There was, however, a 34% increase in glycosaminoglycans and a 20.7% decrease in cell density within the posterior sclera of myopic eyes. The biosynthesis of scleral proteoglycans was determined by measuring 35SO4 incorporation in the sclera of chicks visually occluded for 5, 10, and 15 days. No differences could be detected in 35SO4 incorporation into the cornea or the anterior sclera. However, 35SO4 incorporation was significantly increased in the posterior sclera of myopic eyes by 64% at Day 5, 39% at Day 10, and 49% at Day 15. When fractionated on Sepharose CL-4B, scleral proteoglycans were resolved into two peaks which were identified by Western blot analysis as aggrecan (cartilage proteoglycan) and decorin. Furthermore, Western blot and dot blot analyses indicated that significantly more aggrecan core protein was present in the sclera of myopic eyes compared with equivalent amounts of sclera from control eyes. These results indicate that increased synthesis and accumulation of aggrecan, which increases the volume of extracellular matrix in the posterior sclera, are responsible for the ocular enlargement observed in this model of myopia.     

Does the perception of moving eyes trigger reflexive visual orienting in autism?

Does movement of the eyes in one or another direction function as an automatic attentional cue to a location of interest? Two experiments explored the directional movement of the eyes in a full face for speed of detection of an aftercoming location target in young people with autism and in control participants. Our aim was to investigate whether a low-level perceptual impairment underlies the delay in gaze following characteristic of autism. The participants'' task was to detect a target appearing on the left or right of the screen either 100 ms or 800 ms after a face cue appeared with eyes averting to the left or right. Despite instructions to ignore eye-movement in the face cue, people with autism and control adolescents were quicker to detect targets that had been preceded by an eye movement cue congruent with target location compared with targets preceded by an incongruent eye movement cue. The attention shifts are thought to be reflexive because the cue was to be ignored, and because the effect was found even when cue-target duration was short (100 ms). Because (experiment two) the effect persisted even when the face was inverted, it would seem that the direction of movement of eyes can provide a powerful (involuntary) cue to a location.     

Depth-Dependent Changes in Collagen Organization in the Human Peripapillary Sclera

PurposeThe collagen structure of the human peripapillary sclera plays a significant role in determining optic nerve head (ONH) biomechanics, and is therefore of interest in the study of glaucoma. The aim of the current work was to map the anisotropic collagen structure of the normal human peripapillary sclera as a function of tissue depth.MethodsWide-angle x-ray scattering was used to quantify collagen fibril orientation at 0.5mm intervals across six 150μm-thick serial sections through the peripapillary sclera of eight normal European-derived human eyes. Two structural parameters were measured: 1) the relative number of fibrils preferentially aligned at a given angle within the tissue plane, 2) the degree of collagen alignment (anisotropy).ResultsThe inner-most one-third of the peripapillary scleral stroma (nearest to the choroid) was characterised by collagen fibrils either randomly arranged or preferentially aligned radially with respect to the ONH. In contrast, the outer two-thirds of the tissue was dominated by a circumferential arrangement of collagen encircling the ONH. In all tissue regions the degree of collagen anisotropy peaked in the mid-stroma and progressively decreased towards the tissue surfaces, with the largest depth variations occurring in the inferior-nasal quadrant, and the smallest occurring in the superior-nasal quadrant.ConclusionsSignificant, region-specific variations in collagen structure are present in the human peripapillary sclera as a function of depth. In normal eyes, the circumferential collagen fibril architecture is most prominent in the outer two-thirds of the stroma, possibly as a mechanical adaption to more effectively support the lamina cribrosa at the level of its insertion into the scleral canal wall.     

Humans do not perceive conspecifics with a greater exposed sclera as more trustworthy: a preliminary cross-ethnic study of the function of the overexposed human sclera

Understanding the adaptive function of the unique morphology of the human eye, in particular its overexposed white sclera, may have profound implications for the fields of evolutionary behavioural science, and specifically the areas of human interaction and social cognition. Existing hypotheses, such as the cooperative eye hypothesis, have attracted a lot of attention but remain untested. Here, we: (i) analysed variation in the visible sclera size in humans from different ethnic backgrounds and (ii) examined whether intraspecific variation of exposed sclera size is related to trust. We used 596 facial photographs of men and women, assessed for perceived trustworthiness, from four different self-declared racial backgrounds. The size of the exposed sclera was measured as the ratio between the width of the exposed eyeball and the diameter of the iris (sclera size index, SSI). The SSI did not differ in the four examined races and was sexually monomorphic except for Whites, where males had a larger SSI than females. In general, the association between the SSI and trustworthiness was statistically insignificant. An inverted U-shaped link was found only in White women, yet the strength of the effect of interaction between sex and race was very small. Our results did not provide evidence for the link between exposed sclera size and trustworthiness. We conclude that further investigation is necessary in order to properly assess the hypotheses relating to the socially relevant functions of overexposed sclera.     

The use of autologous fibrin as a scaffold for cultivating autologous conjunctiva in the treatment of conjunctival defect

The purpose of this study was to compare the use of autologous fibrin to human amniotic membrane (HAM) as a scaffold in cultivating autologous conjunctiva for transplantation in treatment of conjunctival defect. An experimental study was performed using 18 adult New Zealand white strain rabbits which were divided into 3 groups. Each group consists of 6 rabbits. The conjunctiva on the temporal site was excised to create a conjunctival epithelial defect. The excised area in the Group 1 was transplanted with autologous conjunctiva cultivated on autologous fibrin; Group 2 was transplanted with autologous conjunctiva cultivated on HAM and Group 3 was left bare. The rabbits were followed up at regular intervals until 6 weeks. The mean period of complete conjunctival epithelization was 11.50 ± 8.22 days for the autologous fibrin group, 15.33 ± 11.80 days for the HAM group and 25.33 ± 5.32 days in the bare sclera group. The epithelization rate for the autologous fibrin group was faster compared to the other two groups. However all the results were not statistically significant (p value >0.05). There were no postoperative complications noted during the follow up. Autologous fibrin is comparable to HAM as a scaffold for cultivation of conjunctiva in the treatment of conjunctival defect.     

Scleral Micro-RNA Signatures in Adult and Fetal Eyes

Introduction

In human eyes, ocular enlargement/growth reflects active extracellular matrix remodeling of the outer scleral shell. Micro-RNAs are small non-coding RNAs that regulate gene expression by base pairing with target sequences. They serve as nodes of signaling networks. We hypothesized that the sclera, like most tissues, expresses micro-RNAs, some of which modulate genes regulating ocular growth. In this study, the scleral micro-RNA expression profile of rapidly growing human fetal eyes was compared with that of stable adult donor eyes using high-throughput microarray and quantitative PCR analyses.

Methods

Scleral samples from normal human fetal (24 wk) and normal adult donor eyes were obtained (n=4 to 6, each group), and RNA extracted. Genome-wide micro-RNA profiling was performed using the Agilent micro-RNA microarray platform. Micro-RNA target predictions were obtained using Microcosm, TargetScan and PicTar algorithms. TaqMan® micro-RNA assays targeting micro-RNAs showing either highest significance, detection, or fold differences, and collagen specificity, were applied to scleral samples from posterior and peripheral ocular regions (n=7, each group). Microarray data were analyzed using R, and quantitative PCR data with 2^-deltaCt methods.

Results

Human sclera was found to express micro-RNAs, and comparison of microarray results for adult and fetal samples revealed many to be differentially expressed (p<0.01, min p= 6.5x10¹¹). Specifically, fetal sclera showed increased expression of mir-214, let-7c, let-7e, mir-103, mir-107, and mir-98 (1.5 to 4 fold changes, p<0.01). However, no significant regionally specific differences .i.e., posterior vs. peripheral sclera, were observed for either adult or fetal samples.

Conclusion

For the first time, micro-RNA expression has been catalogued in human sclera. Some micro-RNAs show age-related differential regulation, higher in the sclera of rapidly growing fetal eyes, consistent with a role in ocular growth regulation. Thus micro-RNAs represent potential targets for ocular growth manipulation, related to myopia and/or other disorders such as scleral ectasia.