Ocular Physiology Research Articles

Adrianus Spigelius' (1578 - 1625) Ocular Anatomy.

The aim was to study Adriaan van den Spiegel's ideas on ocular anatomy. He is better known by his Latinized name as Adrianus Spigelius (1578 - 1625). He was a Flemish physician and anatomist who lived and worked in Padua, where in 1605 he was elected to be Professor of Anatomy and Surgery. Chapter IX of book ten of Spigelius' work on human anatomy, entitled De humani corporis fabrica libri X tabulis aere icisis exornati (1627) was devoted to an anatomical description of the eye. Corresponding to contemporary ideas of the production of knowledge Spigelius endeavoured to enhance Andreas Vesalius' (1514-1564) anatomy, he did not repeat his predecessor's theories of ocular anatomy. He conceptualised that the eye has six muscles, five tunics and three humors, while he gave a brief description of ocular physiology combining anatomy and the functional role of the anatomic ocular parts. CONCLUSION: He managed to correct Vesalius' errors and to present ocular anatomy with original notes, which so far, have been ignored and are highlighted now.

HINTS IN THE ER- case report

Vertigo is a false sense of motion of either the environment or self and is diagnosed in approximately half of the patients with dizziness. Acute spontaneous onset of vertigo is called acute vestibular neuritis (AVN). It is caused by peripheral lesion and requires symptomatic treatment. The symptoms of AVN can mimic a central pathology like cerebellar or brainstem infarction with no concomitant red-flag manifestation. Magnetic Resonance Neuroimaging with Diffusion-weighted imaging (MRI-DWI) as well as Computed Tomography (CT) scan delivers false negatives results what significantly delays stroke treatment. HINTS is an acronym for the battery of three bedside tests of ocular motor physiology. The method is more sensitive in diagnosing posterior circulation infarct than MRI-DWI with specificity -96 %. We present a case of a patient with vertigo who underwent two cranial CT scans and neurological examination. HINTS was worrisome. The brainstem infarct diagnosis was confirmed by MRI-DWI.

Fast versus gradual adaptation of soft daily disposable contact lenses in neophyte wearers

PurposeDespite the widespread practice of gradually adapting all new soft contact lens wearers (neophytes), there is little evidence-based research underpinning such practice. This work determined if a gradual adaptation period is necessary for neophytes when fitted with modern hydrogel or silicone-hydrogel daily disposable contact lenses. MethodAt four sites, neophytes (19–32 years) were randomly assigned to an adaptation schedule: fast (10 h wear from the first day) or gradual (4 h on the first day, increasing their wear-time by 2 h on each subsequent day until they had reached 10 h) with hydrogel (n = 24 fast; n = 21 gradual) or silicone-hydrogel (n = 10 fast; n = 10 gradual) contact lenses. Masked investigators graded ocular surface physiology and non-invasive tear breakup time (NIBUT). A range of subjective scores (using 0–100 visual analogue scales) were recorded at the initial visit and after 10 h of lens wear, 4–6 days and 12–14 days after initial fitting. Subjective scores were also repeated after 7 days. ResultsThere was no difference (p > 0.05) in ocular surface physiology between the fast and gradual adaptation groups at any time point in either lens type. NIBUT was similar at all time points for both adaptation groups in both lens types with the exception that the gradual adaptation silicone-hydrogel wearers had a slightly longer NIBUT (p = 0.007) than the fast adaptation group at 12-14 days. Subjective scores were also similar across the visits and lens types with the exception of ‘lens awareness’ and ‘ease of lens removal’ which were better (p < 0.05) in the fast compared with the gradual adaptation hydrogel lens group at day 7. Additionally, ‘end-of-day discomfort’ was better (p = 0.02) in the fast compared with the gradual adaptation hydrogel lens group at 12–14 days. ConclusionThere appears to be no benefit in daily disposable soft contact lens adaptation for neophytes with modern contact lens materials.

Mathematical modeling highlights from ARVO 2018

At the ARVO annual meeting, there is an increasing number of contributions that involve significant mathematical modeling of ocular physiology and procedures. There has long been significant use of statistical methods for understanding data from a variety of in vivo measurements and clinical trials. Beyond these important uses of statistical and mathematical tools, a growing number of researchers are developing mathematical and computational models, often based on fundamental principles from physics, chemistry and mechanics, that provide insights into ocular phenomena. A number of areas had noticeable contributions involving applications of models, such as tear production, tear film dynamics, corneal biomechanics, retinal blood flow, and glaucoma. We list a number of such contributions in this introduction and follow those with five extended abstracts that summarize some of the studies mentioned here.

On the pivotal role of Elovl3/ELOVL3 in meibogenesis and ocular physiology of mice.

The purpose of this study was to examine the role of Elovl3 gene in meibogenesis and the impact of ELOVL3 protein ablation on the physiology of the mouse ocular surface and Meibomian glands (MGs). Elovl3 knockout, ELOVL3-ablated (E3hom) mice and their wild type littermates (E3wt) were studied side by side. E3hom mice had abnormal ocular phenotypes such as delayed eye opening, weeping eyes, crusty eyelids, eyelid edema, highly vascularized cornea and tarsal plates (TPs), slit eye, and increased tearing that resemble symptoms observed in human subjects with various forms of dry eye, MG dysfunction and blepharitis. Lipid profiling of E3hom TPs was conducted using chromatography and mass spectrometry. The analyses revealed that the lipid composition of E3hom TPs was strikingly different from that of their E3wt littermates. The mutation affected major classes of meibomian lipids - cholesteryl esters, wax esters, and cholesteryl esters of (O)-acylated w-hydroxy fatty acids. The studies illuminated the central role of ELOVL3 in producing C21:0-C29:0 fatty acids, including odd-chain and branched ones. Ablation of ELOVL3 leads to selective changes in the lipid composition of meibum, making E3hom mice instrumental in studying the mechanisms of the biosynthesis of meibum and modeling various pathologies of human ocular surface and adnexa.-Butovich, I. A., Wilkerson, A., Bhat, N., McMahon, A., Yuksel, S. On the pivotal role of Elovl3/ELOVL3 in meibogenesis and ocular physiology of mice.

Contact Lens Technology: From Fundamentals to Applications.

Contact lenses are ocular prosthetic devices used by over 150 million people worldwide. Primary applications of contact lenses include vision correction, therapeutics, and cosmetics. Contact lens materials have significantly evolved over time to minimize adverse effects associated with contact lens wearing, to maintain a regular corneal metabolism, and to preserve tear film stability. This article encompasses contact lens technology, including materials, chemical and physical properties, manufacturing processes, microbial contamination, and ocular complications. The function and the composition of the tear fluid are discussed to assess its potential as a diagnostic media. The regulatory standards of contact lens devices with regard to biocompatibility and contact lens market are presented. Future prospects in contact lens technology are evaluated, with particular interest given to theranostic applications for in situ continuous monitoring the ocular physiology.

Assessment of Zebrafish Lens Nucleus Localization and Sutural Integrity.

The zebrafish is uniquely suited to genetic manipulation and in vivo imaging, making it an increasingly popular model for reverse genetic studies and for generation of transgenics for in vivo imaging. These unique capabilities make the zebrafish an ideal platform to study ocular lens development and physiology. Our recent findings that an Aquaporin-0, Aqp0a, is required for stability of the anterior lens suture, as well as for the shift of the lens nucleus to the lens center with age led us to develop tools especially suited to analyzing the properties of zebrafish lenses. Here we outline detailed methods for lens dissection that can be applied to both larval and adult lenses, to prepare them for histological analysis, immunohistochemistry and imaging. We focus on analysis of lens suture integrity and cortical cell morphology and compare data generated from dissected lenses with data obtained from in vivo imaging of lens morphology made possible by a novel transgenic zebrafish line with a genetically encoded fluorescent marker. Analysis of dissected lenses perpendicular to their optical axis allows quantification of the relative position of the lens nucleus along the anterior-posterior axis. Movement of the lens nucleus from an initial anterior position to the center is required for normal lens optics in adult zebrafish. Thus, a quantitative measure of lens nuclear position directly correlates with its optical properties.

Why does the zebrafish cloche mutant develop lens cataract?

The zebrafish has become a valuable model for examining ocular lens development, physiology and disease. The zebrafish cloche mutant, first described for its loss of hematopoiesis, also shows reduced eye and lens size, interruption in lens cell differentiation and a cataract likely caused by abnormal protein aggregation. To facilitate the use of the cloche mutant for studies on cataract development and prevention we characterized variation in the lens phenotype, quantified changes in gene expression by qRT-PCR and RNA-Seq and compared the ability of two promoters to drive expression of introduced proteins into the cloche lens. We found that the severity of cloche embryo lens cataract varied, while the decrease in lens diameter and retention of nuclei in differentiating lens fiber cells was constant. We found very low expression of both αB-crystallin genes (cryaba and cryabb) at 4 days post fertilization (dpf) by both qRT-PCR and RNA-Seq in cloche, cloche sibling and wildtype embryos and no significant difference in αA-crystallin (cryaa) expression. RNA-Seq analysis of 4 dpf embryos identified transcripts from 25,281 genes, with 1,329 showing statistically significantly different expression between cloche and wildtype samples. Downregulation of eight lens β- and γM-crystallin genes and 22 retinal related genes may reflect a general reduction in eye development and growth. Six stress response genes were upregulated. We did not find misregulation of any known components of lens development gene regulatory networks. These results suggest that the cloche lens cataract is not caused by loss of αA-crystallin or changes to lens gene regulatory networks. Instead, we propose that the cataract results from general physiological stress related to loss of hematopoiesis. Our finding that the zebrafish αA-crystallin promoter drove strong GFP expression in the cloche lens demonstrates its use as a tool for examining the effects of introduced proteins on lens crystallin aggregation and cataract prevention.

Age-related Autoimmune Changes in Lacrimal Glands.

Aging is a complex process associated with dysregulation of the immune system and low levels of inflammation, often associated with the onset of many pathologies. The lacrimal gland (LG) plays a vital role in the maintenance of ocular physiology and changes related to aging directly affect eye diseases. The dysregulation of the immune system in aging leads to quantitative and qualitative changes in antibodies and cytokines. While there is a gradual decline of the immune system, there is an increase in autoimmunity, with a reciprocal pathway between low levels of inflammation and aging mechanisms. Elderly C57BL/6J mice spontaneously show LGs infiltration that is characterized by Th1 but not Th17 cells. The aging of the LG is related to functional alterations, reduced innervation and decreased secretory activities. Lymphocytic infiltration, destruction, and atrophy of glandular parenchyma, ductal dilatation, and secretion of inflammatory mediators modify the volume and composition of tears. Oxidative stress, the capacity to metabolize and eliminate toxic substances decreased in aging, is also associated with the reduction of LG functionality and the pathogenesis of autoimmune diseases. Although further studies are required for a better understanding of autoimmunity and aging of the LG, we described anatomic and immunology aspects that have been described so far.

Student acceptance of e-learning methods in the laboratory class in Optometry.

Today’s students have increased expectations for flexible learning options and evidence-based practice resources to be available to support curricular activities. We investigated: (i) the suitability of a static website for teaching ocular anatomy and physiology and an interactive version of the website with quiz and self-assessment activities and (ii) the usefulness of a blended online and in-lab environment to teach in Optometry. We administered a survey to compare responses of optometry students who had access to the interactive website, with those from students from a previous year who used the static version. We examined learning preferences of students in a focus group. Students were positive about the value of the website for their learning and the clarity of the website content. Nevertheless, objective comparison of pass rates for students using the static and interactive websites did not show significant changes. The majority of students commenting on the static website felt they did not get sufficient feedback via the website (67%) compared with only 22% from students who used self-assessments in the interactive website. Interestingly, users of the static website commented that it was perceived as just another resource while users of the interactive website commented on the usefulness of the material to review knowledge before laboratories. In the focus group, students reported they preferred a blended learning over the website alone even by students using the interactive website as they felt the need to revise content with the educator before the test. We conclude that there is acceptance of online learning methods due to the technologically ‘savvy’ environment of students in the first year of the Optometry programme but there is still dependence on the educator as the main administrator of their learning.

Past and Present of Eye Movement Abnormalities in Ataxia-Telangiectasia.

Ataxia-telangiectasia is the second most common autosomal recessive hereditary ataxia, with an estimated incidence of 1 in 100,000 births. Besides ataxia and ocular telangiectasias, eye movement abnormalities have long been associated with this disorder and is frequently present in almost all patients. A handful of studies have described the phenomenology of ocular motor deficits in ataxia-telangiectasia. Contemporary literature linked their physiology to cerebellar dysfunction and secondary abnormalities at the level of brainstem. These studies, while providing a proof of concept of ocular motor physiology in disease, i.e., ataxia-telangiectasia, also advanced our understanding of how the cerebellum works. Here, we will summarize the clinical abnormalities seen with ataxia-telangiectasia in each subtype of eye movements and subsequently describe the underlying pathophysiology. Finally, we will review how these deficits are linked to abnormal cerebellar function and how it allows better understanding of the cerebellar physiology.

Challenges in the Polyene- and Azole-Based Pharmacotherapy of Ocular Fungal Infections.

Polyenes and azoles constitute 2 major drug classes in the antifungal armamentarium used to treat fungal infections of the eye such as fungal keratitis, endophthalmitis, conjunctivitis, and blepharitis. These classes of drugs have come to occupy an important niche in ophthalmic antifungal therapy due to their broad spectrum of activity against a variety of filamentous and yeast-like fungi. Natamycin suspension (Natacyn®), a polyene antifungal drug, is currently the only US FDA-approved formulation for treating ophthalmic fungal infections, whereas the other polyene and azole antifungals such as amphotericin B, fluconazole, itraconazole, ketoconazole, miconazole, voriconazole, and posaconazole are routinely used off-label in the clinical setting. Despite potent antifungal activity, the clinical utility of these agents in ophthalmic infections has been challenged by their physicochemical properties, the unique ocular anatomy and physiology, selective antifungal activity, ocular and systemic toxicity, emergence of resistance and cross-resistance, and absence of reliable techniques for developing a robust in vitro-in vivo correlation. This review discusses the aforementioned challenges and the common approaches undertaken to circumnavigate the difficulties associated with the polyene- and azole-based pharmacotherapy of ophthalmic fungal infections.

Remodeling of Retinal Architecture in Diabetic Retinopathy: Disruption of Ocular Physiology and Visual Functions by Inflammatory Gene Products and Pyroptosis.

Diabetic patients suffer from a host of physiological abnormalities beyond just those of glucose metabolism. These abnormalities often lead to systemic inflammation via modulation of several inflammation-related genes, their respective gene products, homocysteine metabolism, and pyroptosis. The very nature of this homeostatic disruption re-sets the overall physiology of diabetics via upregulation of immune responses, enhanced retinal neovascularization, upregulation of epigenetic events, and disturbances in cells’ redox regulatory system. This altered pathophysiological milieu can lead to the development of diabetic retinopathy (DR), a debilitating vision-threatening eye condition with microvascular complications. DR is the most prevalent cause of irreversible blindness in the working-age adults throughout the world as it can lead to severe structural and functional remodeling of the retina, decreasing vision and thus diminishing the quality of life. In this manuscript, we attempt to summarize recent developments and new insights to explore the very nature of this intertwined crosstalk between components of the immune system and their metabolic orchestrations to elucidate the pathophysiology of DR. Understanding the multifaceted nature of the cellular and molecular factors that are involved in DR could reveal new targets for effective diagnostics, therapeutics, prognostics, preventive tools, and finally strategies to combat the development and progression of DR in susceptible subjects.

Identification of unique proteins in vitreous fluid of patients with noninfectious uveitis.

Uveitis is a cause for concern in the developing countries like India. Its poor diagnosis and lack of proper therapeutics often cause blindness in children and young adults. Moreover, the exact mechanism of pathogenesis of different types of uveitis is still elusive. Modern proteomic techniques are found to be advantageous for an in-depth understanding of the ocular physiology using proteomic diversity. Our aim was to identify unique proteins involved in the pathogenesis of autoimmune or noninfectious uveitis. Vitreous fluid samples (n=90) were obtained from infectious (N=34) and noninfectious (N=56) uveitis patients, and their protein profiles were compared by analysing sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 2D electrophoresis. Unique proteins were identified through matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and further studied for pathway analysis. Protein spots having different molecular weights were observed in noninfectious vitreous fluid samples. Enzymatic digestion of these spots after MALDI-TOF MS analysis revealed different proteins. We identified 25 different proteins through SDS-PAGE and 22 through 2D electrophoresis. 50% of the proteins from SDS-PAGE were associated with heterotrimeric G-protein signalling pathway-rod outer segment phototransduction. 50% proteins from SDS-PAGE and 20% from 2D electrophoresis revealed association with de novo purine biosynthesis. Carbonic anhydrase 1 and serpin B3 were found to be common in both analyses. High-throughput proteomic and pathway analyses have exposed the potential association of these proteins with autoimmune pathogenesis in uveitis. The exact role of most of the proteins in autoimmune uveitis is yet to be unfurled.

Novel drug delivery systems for ocular therapy: With special reference to liposomal ocular delivery.

Delivery of drugs to eyes is a great challenge to researchers because of a number of barriers in the eye preventing the actual dose from reaching the site. A number of ophthalmic delivery systems have been developed in the past couple of years that are not only new but also safe and reliable and help to overcome all those barriers in the eye which are responsible for the very less bioavailability of drugs. In this review, we tried to focus on current research in ocular delivery of drug substances giving special emphasis to liposomal delivery system. A brief analysis of other novel ocular delivery systems, ocular physiology, and microbial sources of disease are also highlighted herein. We analyzed the various research findings for churning a general idea for novel ocular delivery system and its future use. The novel formulations may overcome the addressed problems of ophthalmic medication and comply with the quality assurance issues. The liposomal delivery is advantageous as they have the ability to entrap both hydrophobic and hydrophilic drugs and are suitable for delivery to both the anterior and posterior segment of the eye. Therefore, the use of this alternative approach is quite a necessity.

Clinical comparison of optimum and large diameter soft contact lenses.

To compare the clinical performance of large diameter lenses with optimally fit lenses in the same material and monocurve back surface design. In a four-visit, randomised, bilateral, crossover, study, 25 myopic subjects wore optimum diameter lenses (control) and large diameter lenses (test) in random succession for 1 week each. Both study lenses were made of methafilcon A and of an identical design. Trial fittings with Frequency 55 (Coopervision) lenses modified with a design algorithm were used to determine the appropriate custom-made study lenses. The least squares mean scores (±SE) for overall comfort and end-of-day comfort (0-10 scale) were 7.57 ± 0.33 vs. 7.42 ± 0.33 (P = 0.59) and 7.00 ± 0.31 vs. 7.27 ± 0.32 (P > 0.05) for the optimum and large diameter lenses, respectively. There were no significant differences in mean (±SE) gradings for limbal hyperaemia (1.23 ± 0.11 vs. 1.19 ± 0.11, 0-4 scale, P = 0.60) and corneal staining (1.79 ± 0.25 vs. 2.04 ± 0.25, P = 0.39). Conjunctival staining was greater for the optimum lens: 1.80 ± 0.28 vs. 0.93 ± 0.28 (0-4 scale, P = 0.001). With regard to lens fit, the large diameter lenses showed significantly less post-blink movement (0.22 ± 0.01 vs. 0.16 ± 0.01 mm, P = 0.004), and greater total decentration (0.15 ± 0.02 vs. 0.21 ± 0.02 mm, P = 0.010). However, there was no significant difference in the key fit variable of tightness on push-up (46 ± 0.69% vs. 48 ± 0.69%, 0-100 scale, P = 0.12). The findings suggest that larger than optimal soft lenses may be worn without detriment to either comfort or ocular physiology, provided an optimal fit is otherwise maintained.

Age-related Changes in Eye, Brain and Visuomotor Behavior in the DBA/2J Mouse Model of Chronic Glaucoma

Although elevated intraocular pressure (IOP) and age are major risk factors for glaucoma, their effects on glaucoma pathogenesis remain unclear. This study examined the onset and progression of glaucomatous changes to ocular anatomy and physiology, structural and physiological brain integrity, and visuomotor behavior in the DBA/2J mice via non-invasive tonometry, multi-parametric magnetic resonance imaging (MRI) and optokinetic assessments from 5 to 12 months of age. Using T2-weighted MRI, diffusion tensor MRI, and manganese-enhanced MRI, increasing IOP elevation at 9 and 12 months old coincided with anterior chamber deepening, altered fractional anisotropy and radial diffusivity of the optic nerve and optic tract, as well as reduced anterograde manganese transport along the visual pathway respectively in the DBA/2J mice. Vitreous body elongation and visuomotor function deterioration were observed until 9 months old, whereas axial diffusivity only decreased at 12 months old in diffusion tensor MRI. Under the same experimental settings, C57BL/6J mice only showed modest age-related changes. Taken together, these results indicate that the anterior and posterior visual pathways of the DBA/2J mice exhibit differential susceptibility to glaucomatous neurodegeneration observable by in vivo multi-modal examinations.

Daily versus monthly disposable contact lens: Which is better for ocular surface physiology and comfort?

To investigate the effect of soft contact lenses (SCL) wearing modality and lens materials on the changes in conjunctival bulbar and limbal redness and conjunctival and corneal staining after two months of SCL wear. Comfort level was also analyzed. In this longitudinal clinical trial, forty-seven neophyte myopic subjects were fitted with a monthly disposable lens (lotrafilcon-B or comfilcon-A or balafilcon-A) in one eye and a daily disposable lens (nelfilcon-A or stenofilcon-A or nesofilcon-A) in the other eye, randomly selected. Conjunctival bulbar and limbal redness and conjunctival and corneal staining were evaluated before and after lens wear. Effect of lens wearing modality and lens materials on these changes was also determined. Level of comfort was evaluated subjectively twice per day. Comfort level and reduction in end-of-day comfort were compared between different lens wearing modalities and materials. Bulbar and limbal redness and conjunctival and corneal staining were increased (p<0.001) after lens wear, and changes were similar with daily and monthly disposable lens wear (p>0.05). Limbal redness was associated with lens materials, and lotrafilcon-B induced the least among the studied lenses (p<0.05). There was no significant association between the wearing modality and the average comfort level and reduction of end-of-day comfort (p>0.05). Two months of SCL wear increased conjunctival redness, conjunctival and corneal staining, which were not associated with the lens wearing modality. There was a reduction in end-of-day comfort, similar to daily and monthly lenses. The change in limbal redness and reduction in end-of-day comfort were associated with the characteristics of the lens material.

Subretinal and Intravitreal Retinal Injections in Monkeys.

Ocular anatomy and physiology in cynomolgus monkeys (Macacca fascicularis) are very similar to that found in the human visual system. Therefore and despite significant ethical and economical implications, these animals constitute an excellent model system for toxicology and biodistribution studies in the development of new and meaningful treatment strategies for ocular disorders. The methods for delivery of investigational new drugs (INDs) to the ocular tissue are virtually identical to the methods used in clinical practice. This protocol explains in detail the method of applying INDs to the vitreous cavity or the subretinal space in monkeys.

Cold-sensing TRPM8 channel participates in circadian control of the brown adipose tissue

Transient receptor potential (TRP) channels are known to regulate energy metabolism, and TRPM8 has become an interesting player in this context. Here we demonstrate the role of the cold sensor TRPM8 in the regulation of clock gene and clock controlled genes in brown adipose tissue (BAT). We investigated TrpM8 temporal profile in the eyes, suprachiasmatic nucleus and BAT; only BAT showed temporal variation of TrpM8 transcripts. Eyes from mice lacking TRPM8 lost the temporal profile of Per1 in LD cycle. This alteration in the ocular circadian physiology may explain the delay in the onset of locomotor activity in response to light pulse, as compared to wild type animals (WT). Brown adipocytes from TrpM8 KO mice exhibited a larger multilocularity in comparison to WT or TrpV1 KO mice. In addition, Ucp1 and UCP1 expression was significantly reduced in TrpM8 KO mice in comparison to WT mice. Regarding circadian components, the expression of Per1, Per2, Bmal1, Pparα, and Pparβ oscillated in WT mice kept in LD, whereas in the absence of TRPM8 the expression of clock genes was reduced in amplitude and lack temporal oscillation. Thus, our results reveal new roles for TRPM8 channel: it participates in the regulation of clock and clock-controlled genes in the eyes and BAT, and in BAT thermogenesis. Since disruption of the clock machinery has been associated with many metabolic disorders, the pharmacological modulation of TRPM8 channel may become a promising therapeutic target to counterbalance weight gain, through increased thermogenesis, energy expenditure, and clock gene activation.