Publications by authors named "Tor P Utheim"

61 Publications

Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases.

Front Immunol 2021 22;12:618653. Epub 2021 Apr 22.

Schepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United States.

The amount of mucin secreted by conjunctival goblet cells is regulated to ensure the optimal level for protection of the ocular surface. Under physiological conditions lipid specialized pro-resolving mediators (SPM) are essential for maintaining tissue homeostasis including the conjunctiva. The protein Annexin A1 (AnxA1) can act as an SPM. We used cultured rat conjunctival goblet cells to determine if AnxA1 stimulates an increase in intracellular [Ca] ([Ca]) and mucin secretion and to identify the signaling pathways. The increase in [Ca] was determined using fura2/AM and mucin secretion was measured using an enzyme-linked lectin assay. AnxA1 stimulated an increase in [Ca] and mucin secretion that was blocked by the cell-permeant Ca chelator BAPTA/AM and the ALX/FPR2 receptor inhibitor BOC2. AnxA1 increased [Ca] to a similar extent as the SPMs lipoxin A and Resolvin (Rv) D1 and histamine. The AnxA1 increase in [Ca] and mucin secretion were inhibited by blocking the phospholipase C (PLC) pathway including PLC, the IP receptor, the Ca/ATPase that causes the intracellular Ca stores to empty, and blockade of Ca influx. Inhibition of protein kinase C (PKC) and Ca/calmodulin-dependent protein kinase also decreased the AnxA1-stimulated increase in [Ca] and mucin secretion. In contrast inhibitors of ERK 1/2, phospholipase A (PLA), and phospholipase D (PLD) did not alter AnxA1-stimulated increase in [Ca], but did inhibit mucin secretion. Activation of protein kinase A did not decrease either the AnxA1-stimulated rise in [Ca] or secretion. We conclude that in health, AnxA1 contributes to the mucin layer of the tear film and ocular surface homeostasis by activating the PLC signaling pathway to increase [Ca] and stimulate mucin secretion and ERK1/2, PLA, and PLD to stimulate mucin secretion from conjunctival goblet cells.
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http://dx.doi.org/10.3389/fimmu.2021.618653DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8100605PMC
April 2021

Congenital aniridia - A comprehensive review of clinical features and therapeutic approaches.

Surv Ophthalmol 2021 Mar 4. Epub 2021 Mar 4.

Department of Ophthalmology, Oslo University Hospital, Oslo, Norway; Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.

Congenital aniridia is a rare genetic eye disorder with total or partial absence of the iris from birth. In most cases the genetic origin of aniridia is a mutation in the PAX6 gene, leading to involvement of most eye structures. Hypoplasia of the fovea is usually present and is associated with reduced visual acuity and nystagmus. Aniridia-associated keratopathy, glaucoma, and cataract are serious and progressive complications that can further reduce visual function. Treatment of the ocular complications of aniridia is challenging and has a high risk of side effects. New approaches such as stem cell therapy may, however, offer better prognoses. We describe the various ocular manifestations of aniridia, with a special focus on conditions that commonly require treatment. We also review the growing literature reporting systemic manifestations of the disease.
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http://dx.doi.org/10.1016/j.survophthal.2021.02.011DOI Listing
March 2021

Tear Film Break-Up Time and Dry Eye Disease Severity in a Large Norwegian Cohort.

J Clin Med 2021 Feb 22;10(4). Epub 2021 Feb 22.

The Norwegian Dry Eye Clinic, 0366 Oslo, Norway.

This study evaluated to what extent tear film break-up time (TFBUT) could discriminate pathological scores for other clinical tests and explore the associations between them. Dry eye patients ( = 2094) were examined for ocular surface disease index (OSDI), tear film osmolarity (Osm), TFBUT, blink interval, ocular protection index (OPI), ocular surface staining (OSS), Schirmer I test, meibomian expressibility, meibomian quality, and meibomian gland dysfunction. The results were grouped into eight levels of break-up time (≤2, ≥3, ≤5, ≥6, ≤10, ≥11, ≤15, and ≥16) with or without sex stratification. Receiver-operating characteristic curve (ROC) analysis and Pearson's correlation coefficients were used to study TFBUT's discriminative power and the associations among the tests, respectively. Above and below each TFBUT's cut-off, all of the parameters indicated significant difference between groups, except OSDI (cut-off 15 s) and Osm (cut-offs 5 s-15 s). At TFBUT cut-off of 2 s, sex difference could be detected for OSDI, Osm, and OSS. OPI presented the strongest discriminative power and association with TFBUT in sharp contrast to Osm, holding the poorest discriminative power with no significant correlation. The remaining parameters were within the poor to very poor categories, both with regard to discrimination and correlation. In conclusion, patients with lower TFBUT presented with more severe DED parameters at all four defined cut-off values.
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http://dx.doi.org/10.3390/jcm10040884DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7926845PMC
February 2021

Oral and ocular late effects in head and neck cancer patients treated with radiotherapy.

Sci Rep 2021 Feb 17;11(1):4026. Epub 2021 Feb 17.

Department of Oral Surgery and Oral Medicine, Faculty of Dentistry, University of Oslo, Oslo, Norway.

A broader understanding of oral and ocular late effects in head and neck cancer (HNC) patients who underwent intensity-modulated radiotherapy (IMRT) may provide valuable information in follow-up and improve quality of life. Twenty-nine HNC patients treated at least 6 months earlier and 30 age-matched controls were recruited. After completing several questionnaires: Oral Health Impact Profile-14 (OHIP-14), Shortened Xerostomia Inventory (SXI), Ocular Surface Disease Index (OSDI) and McMonnies Dry Eye questionnaire (MDEQ), participants underwent oral and ocular examinations. Oral examination included clinical oral dryness score (CODS) and secretion rates of unstimulated and stimulated saliva (UWS, SWS). Ocular examination included tear film break-up time, Schirmer test and ocular surface staining. The patients had more problems related to dry mouth than controls based on CODS and SXI, and more complaints of dry eye disease based on OSDI and MDEQ. UWS and SWS rates and oral health related quality of life were significantly lower in the patient group. Subjective oral dryness (SXI) correlated significantly with subjective ocular dryness (OSDI and MDEQ). Our study demonstrates that HNC patients treated with IMRT experience late effects in terms of xerostomia and ocular dryness underlining the importance of interdisciplinary approach in the evaluation and follow-up of HNC patients.
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http://dx.doi.org/10.1038/s41598-021-83635-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7889862PMC
February 2021

STRUCTURAL PATHOLOGY AFTER RETINAL DETACHMENT: MULTICOLOR CONFOCAL SCANNING LASER OPHTHALMOSCOPY VERSUS COLOR FUNDUS PHOTOGRAPHY.

Retina 2021 Jan 8. Epub 2021 Jan 8.

Department of Ophthalmology, Stavanger University Hospital, Stavanger, Norway Department of Quality and Health Technology, University of Stavanger, Stavanger, Norway Department of Research, Stavanger University Hospital, Stavanger, Norway Department of Clinical Medicine, Section of Ophthalmology, University of Bergen, Bergen, Norway Department of Ophthalmology, Haukeland University Hospital, Bergen, Norway Department of Ophthalmology, St. Olavs Hospital, Trondheim University Hospital and Department of Neuromedicine and Movement Science, Norwegian University of Science and Technology, Norway Department of Ophthalmology, University Hospital of Northern Norway, Norway.

Purpose: To compare pathology detection using multicolor confocal scanning laser ophthalmoscopy (MCcSLO) with color fundus photography (CFP) following macula-off rhegmatogenous retinal detachment (RRD).

Methods: Post-surgery MCcSLO and CFP images from 30 RRD patients were examined by 10 masked graders. Imaging was performed with the Heidelberg Spectralis HRA and the digital retinal camera Basler acA2500-14gc GigE. Swept-source optical coherence tomography was used as verification modality. Detection rates of ellipsoid zone disruption, foveal ellipsoid zone rosette, outer retinal folds, intraretinal cysts, subretinal fluid layer, subretinal fluid blebs, retinal striae and retinal detachment line were compared. Inter-modality and inter-grader agreement were estimated.

Results: Overall pathology detection was significantly higher for MCcSLO (aOR=7.39 (95% CI, 1.64-33.30); P=0.009). The inter-modality and inter-grader agreement on overall pathology detection were moderate. The inter-modality agreement was 0.49 (95% CI, 0.48-0.51; P <0.0001) (Gwet's AC1). Inter-grader agreement was 0.53 (95% CI, 0.52-0.54; P <0.0001) for MCcSLO and 0.58 (95% CI, 0.57-0.59; P <0.0001) (Fleiss kappa) for CFP.

Conclusions: MCcSLO imaging is superior to CFP in detecting and delineating structural retinal pathologies following RRD, and can be a helpful tool in the visualization of retinal remodeling processes in patients recovering from RRD surgery.
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http://dx.doi.org/10.1097/IAE.0000000000003113DOI Listing
January 2021

Non-invasive electrical stimulation as a potential treatment for retinal degenerative diseases.

Neural Regen Res 2021 Aug;16(8):1558-1559

Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA.

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http://dx.doi.org/10.4103/1673-5374.303015DOI Listing
August 2021

Sex and age differences in symptoms and signs of dry eye disease in a Norwegian cohort of patients.

Ocul Surf 2021 01 24;19:68-73. Epub 2020 Nov 24.

National Centre for Vision, Optics and Eye Care, Department of Optometry, Radiography and Lighting Design, Faculty of Health and Social Sciences, University of South-Eastern Norway, Norway; Department of Ophthalmology, Vestre Viken Hospital Trust, Drammen, Norway; Department of Ophthalmology, Sørlandet Hospital Arendal, Arendal, Norway; The Norwegian Dry Eye Clinic, Oslo, Norway; Department of Oral Surgery and Oral Medicine, Faculty of Dentistry, University of Oslo, Oslo, Norway.

Purpose: To investigate sex and age differences in symptoms and signs in a Norwegian clinic-based cohort of patients with dry eye disease (DED).

Methods: Visitors at the Norwegian Dry Eye Clinic were examined using Ocular Surface Disease Index (OSDI) questionnaire score, tear osmolarity, tear break-up time (TFBUT), ocular surface staining, corneal sensitivity, Schirmer I test, and meibum expressibility (ME) and quality (MQ). A diagnosis of DED was made by an ophthalmologist based on symptoms and signs, and only DED patients were enrolled in the study: 1823 patients (338 males; mean age 51.2 ± 16.2 years; 1485 females; mean age 52.5 ± 16.0 years). The patients were divided into age subgroups: 20-39 years, 40-59 years and ≥60 years. Sex differences in the aforementioned tests were analyzed. Values were reported as mean ± standard deviation (SD), and intergroup comparisons were performed using Mann-Whitney U test. Multiple regression was used to analyze sex and age influences on symptoms and signs.

Results: When patients of all ages were analyzed, females had increased osmolarity, shorter TFBUT, reduced MQ and ME and higher corneal sensitivity. OSDI, Schirmer I test, ocular surface staining and corneal staining were not significantly different between the sexes. Only with TFBUT and ME were the sex difference present in all age subgroups. Multiple regression showed that all parameters were influenced by either sex or age, but only TFBUT and ME were influenced by both sex and age. (all p < 0.05).

Conclusions: Sex and age differences in dry eye were most consistent in TFBUT and ME, that indicate differences in meibomian gland functionality. Sex and age subgroup stratification is important in future studies investigating DED in other populations.
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http://dx.doi.org/10.1016/j.jtos.2020.11.009DOI Listing
January 2021

Advances, limitations and future perspectives in the diagnosis and management of dry eye in Sjögren's syndrome.

Clin Exp Rheumatol 2020 Jul-Aug;38 Suppl 126(4):301-309. Epub 2020 Sep 25.

Section of Academic Ophthalmology, School of Life Course Sciences, Faculty of Life Course Sciences and Medicine (FoLSM), King's College London, St. Thomas' Hospital Campus, London, and Department of Ophthalmology, Guys and St Thomas' NHS Foundation Trust, St Thomas' Hospital, London, UK.

Primary Sjögren's syndrome is a complex systemic autoimmune disorder that primarily affects exocrine glands such as the lacrimal glands. Dry eye disease is one of the most prevalent complications of Sjögren's syndrome, affecting most patients. It significantly impairs quality of life and management is often difficult and unsatisfactory, in part due to weak correlation between symptoms and signs and poor recognition of the three main subtypes aqueous-deficient, evaporative and neuropathic dry eye. This review provides an overview of key aspects of dry eye disease, such as its multifactorial aetiology and recent insights into pathophysiology. The uses and pitfalls of commonly-used diagnostic tests for dry eye are reviewed, as well as the increasing number of new imaging technologies and biomarkers to refine diagnosis. There are many current and emerging treatment options for dry eye in Sjögren's syndrome, but high-level evidence of efficacy is mostly lacking, as are evidence-based treatment algorithms. All these aspects make the management of dry eye in Sjögren's syndrome challenging.
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October 2020

Povidone iodine treatment is deleterious to human ocular surface conjunctival cells in culture.

BMJ Open Ophthalmol 2020 16;5(1):e000545. Epub 2020 Sep 16.

Schepens Eye Research Institute, Boston, Massachusetts, USA.

Objective: To determine the effect of povidone iodine (PI), an antiseptic commonly used prior to ocular surgery, on viability of mixed populations of conjunctival stratified squamous and goblet cells, purified conjunctival goblet cells and purified conjunctival stromal fibroblasts in primary culture.

Methods And Analysis: Mixed population of epithelial cells (stratified squamous and goblet cells), goblet cells and fibroblasts were grown in culture from pieces of human conjunctiva using either supplemented DMEM/F12 or RPMI. Cell type was evaluated by immunofluorescence microscopy. Cells were treated for 5 min with phosphate-buffered saline (PBS); 0.25%, 2.5%, 5% or 10% PI in PBS; or a positive control of 30% HO. Cell viability was determined using Alamar Blue fluorescence and a live/dead kit using calcein/AM and ethidium homodimer-1 (EH-1).

Results: Mixed populations of epithelial cells, goblet cells and fibroblasts were characterised by immunofluorescence microscopy. As determined with Alamar Blue fluorescence, all concentrations of PI significantly decreased the number of cells from all three preparation types compared with PBS. As determined by calcein/EH-1 viability test, mixed populations of cells and fibroblasts were less sensitive to PI treatment than goblet cells. All concentrations of PI, except for 0.25% used with goblet cells, substantially increased the number of dead cells for all cell populations. The HO control also significantly decreased the number and viability of all three types of cells in both tests.

Conclusion: We conclude that PI, which is commonly used prior to ocular surgeries, is detrimental to human conjunctival stratified squamous cells, goblet cells and fibroblasts in culture.
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http://dx.doi.org/10.1136/bmjophth-2020-000545DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7497553PMC
September 2020

Lacrimal Gland Myoepithelial Cells Are Altered in a Mouse Model of Dry Eye Disease.

Am J Pathol 2020 10 15;190(10):2067-2079. Epub 2020 Jul 15.

Schepens Eye Research Institute/Massachusetts Eye and Ear, Harvard Medical School, Boston, Massachusetts; Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts. Electronic address:

The purpose of this study was to determine the pathogenic changes that occur in myoepithelial cells (MECs) from lacrimal glands of a mouse model of Sjögren syndrome. MECs were cultured from lacrimal glands of C57BL/6J [wild type (WT)] and thrombospondin 1 null (TSP1, alias Thbs1) mice and from mice expressing α-smooth muscle actin-green fluorescent protein that labels MECs. MECs were stimulated with cholinergic and α-adrenergic agonists, vasoactive intestinal peptide (VIP), and the purinergic agonists ATP and UTP. Then intracellular [Ca] was measured using fura-2, and contraction was observed using live cell imaging. Expression of purinergic receptors was determined by Western blot analysis, and mRNA expression was analyzed by microarray. The increase in intracellular [Ca] with VIP and UTP was significantly smaller in MECs from TSP1 compared with WT mice. Cholinergic agonists, ATP, and UTP stimulated contraction in MECs, although contraction of MECs from TSP1 mice was reduced compared with WT mice. The amount of purinergic receptors P2Y1, P2Y11, and P2Y13 was significantly decreased in MECs from TSP1 compared with WT mice, whereas several extracellular matrix and inflammation genes were up-regulated in MECs from TSP1 mice. We conclude that lacrimal gland MEC function is altered by inflammation because the functions regulated by cholinergic agonists, VIP, and purinergic receptors are decreased in TSP1 compared with WT mice.
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http://dx.doi.org/10.1016/j.ajpath.2020.06.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7520664PMC
October 2020

Maresin 1, a specialized proresolving mediator, stimulates intracellular [Ca ] and secretion in conjunctival goblet cells.

J Cell Physiol 2021 Jan 8;236(1):340-353. Epub 2020 Jun 8.

Schepens Eye Research Institute/Massachusetts Eye and Ear, Boston, Massachusetts.

Mucin secretion from conjunctival goblet cells forms the tear film mucin layer and requires regulation to function properly. Maresin 1 (MaR1) is a specialized proresolving mediator produced during the resolution of inflammation. We determined if MaR1 stimulates mucin secretion and signaling pathways used. Cultured rat conjunctival goblet cells were used to measure the increase in intracellular Ca ([Ca ] ) concentration and mucin secretion. MaR1-increased [Ca ] and secretion were blocked by inhibitors of phospholipase C, protein kinase C, Ca /calmodulin-dependent protein kinase II, and extracellular-regulated kinase 1/2. MaR1 added before addition of histamine counterregulated histamine-stimulated increase in [Ca ] and secretion. We conclude that MaR1 likely has two actions in conjunctival goblet cells: first, maintaining optimal tear film mucin levels by increasing [Ca ] and stimulating mucin secretion in health and, second, attenuating the increase in [Ca ] and overproduction of mucin secretion by counterregulating the effect of histamine as occurs in ocular allergy.
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http://dx.doi.org/10.1002/jcp.29846DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7722204PMC
January 2021

The role of meibography in ocular surface diagnostics: A review.

Ocul Surf 2021 01 19;19:133-144. Epub 2020 May 19.

Department of Medical Biochemistry, Oslo University Hospital, Norway; The Norwegian Dry Eye Clinic, Ole Vigs Gate 32 E, 0366, Oslo, Norway; Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Norway; Department of Ophthalmology, Sørlandet Hospital Arendal, Norway; Department of Ophthalmology, Stavanger University Hospital, Norway.

The meibomian glands are lipid-secreting glands located in the tarsal plates, whose secretory products cover the tear film, thereby reducing evaporation as well as ensuring lubrication of the ocular surface. The meibomian glands can be visualized at different levels of magnification by infrared meibography, laser confocal microscopy, and optical coherence tomography. These imaging modalities have been subject to much research and progress in clinical practice and have shaped our current understanding of meibomian glands in health and disease. In this review, we explore the evolution of meibography over the past decades, the major contributions of various meibographic modalities, and discuss their clinical significance.
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http://dx.doi.org/10.1016/j.jtos.2020.05.004DOI Listing
January 2021

Temporal redistribution of cap and residual stromal thickness after SMILE.

J Cataract Refract Surg 2020 Oct;46(10):1331-1338

From the Department of Medical Biochemistry, Oslo University Hospital, Oslo (Olsen, Lyngstadaas, Utheim), Department of Ophthalmology (Utheim), Oslo University Hospital, Oslo, Institute of Clinical Medicine (Olsen, Lyngstadaas), Faculty of Medicine, University of Oslo, Oslo, Department of Ophthalmology (Stojanovic), University Hospital North Norway, Tromsø, Institute of Clinical Medicine (Zhou, Stojanovic), Faculty of Medicine, University of Tromsø, Tromsø, Department of Ophthalmology (Utheim, Chen), Sørlandet Hospital, Arendal, Department of Ophthalmology (Utheim, Chen), Vestre Viken Hospital Trust, Drammen, Faculty of Health Sciences, National Centre for Optics, Vision and Eye Care, University of South Eastern Norway (Utheim, Badian, Chen), Kongsberg, Norway; and the Department of Ophthalmology (Wang), The First Affiliated Hospital of Northwestern University, Xi´an, Shanxi, China.

Purpose: To investigate corneal sublayer alterations during the postoperative period after small-incision lenticule extraction (SMILE).

Setting: Synslaser clinic, Oslo, Norway.

Study Design: Retrospective.

Methods: Patients who underwent SMILE for treating myopia were included. The thicknesses of the corneal epithelium, cap, stromal part of the cap (StromaCap), residual stromal bed (StromaRes), and total stroma (StromaTot) were measured using spectral-domain optical coherence tomography at 1 day, 1 week, 1 month, 3 months, and 6 months postoperatively. Postoperative changes in the corneal sublayer thicknesses were analyzed and correlated with changes in spherical equivalence and anterior and posterior keratometry (K).

Results: The study was based on analyses of the right eyes of 51 patients. From 1 day to 6 months postoperatively, the corneal epithelium, cap, StromaCap, StromaRes, and StromaTot thicknesses increased from 54.4 ± 4.0 µm to 57.3 ± 5.2 µm; 137.1 ± 5.5 µm to 140.3 ± 5.1 µm; 82.7 ± 5.9 µm to 82.8 ± 6.3 µm; 375.0 ± 40.8 µm to 381.4 ± 30.6 µm; and 457.6 ± 41.1 µm to 462.1 ± 36.7 µm, respectively. Between 1 month and 6 months postoperatively, the increase in anterior K correlated significantly with the thickening of the cap (r = 0.37, P = .03) and the stromal component of the cap (r = 0.36, P = .04) within the central cornea.

Conclusions: The post-SMILE remodeling behavior between the anterior (StromaCap) and posterior (StromaRes) stroma were dissimilar. There was a significant correlation between changes in anterior K and the central cap and the stromal component of the cap. This might be because of biomechanical changes, tissue remodeling, and wound healing or a combination of some or all of the aforementioned processes.
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http://dx.doi.org/10.1097/j.jcrs.0000000000000242DOI Listing
October 2020

PAX6 Genotypic and Retinal Phenotypic Characterization in Congenital Aniridia.

Invest Ophthalmol Vis Sci 2020 05;61(5):14

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Purpose: To investigate the association between PAX6 genotype and macular morphology in congenital aniridia.

Methods: The study included 37 participants (15 males) with congenital aniridia (aged 10-72 years) and 58 age-matched normal controls (18 males). DNA was isolated from saliva samples. PAX6 exons, intron/exon junctions, and known regulatory regions were amplified in PCR and sequenced. Multiplex ligation-dependent probe amplification (MLPA) was performed to detect larger deletions or duplications in PAX6 or known cis-regulatory regions. Spectral-domain optical coherence tomography images were acquired and segmented semiautomatically. Mean thicknesses were calculated for inner and outer retinal layers within the macula along nasal and temporal meridians.

Results: Mutations in PAX6 or regulatory regions were found in 97% of the participants with aniridia. Foveal hypoplasia was observed in all who had a mutation within the PAX6 gene. Aniridic eyes had thinner outer retinal layers than controls, but with large between-individual variation (mean ± SD, 156.3 ± 32.3 µm vs 210.8 ± 12.3 µm, P < 0.001). Parafoveal and perifoveal inner and outer retinal layers were thinner in aniridia. Participants with mutations in noncoding PAX6 regions had thicker foveal outer retinal layers than those with mutations in the PAX6 coding regions (P = 0.04) and showed signs of postnatal development and maturation. Mutations outside the PAX6 gene were associated with the mildest retinal phenotypes.

Conclusions: PAX6 mutations are associated with significant thinning of macular inner and outer retinal layers, consistent with misdirected retinal development resulting in abnormal foveal formation and reduced number of neurons in the macula, with mutations in PAX6 coding regions giving the worst outcome.
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http://dx.doi.org/10.1167/iovs.61.5.14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7405610PMC
May 2020

RvE1 uses the LTB receptor BLT1 to increase [Ca] and stimulate mucin secretion in cultured rat and human conjunctival goblet cells.

Ocul Surf 2020 07 30;18(3):470-482. Epub 2020 Apr 30.

Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA. Electronic address:

Purpose: Specialized pro-resolving lipid mediator resolvin (Rv) E1 stimulates secretion including mucins from conjunctival goblet cells. RvE1 can use both its ChemR23 receptor and the LTB receptor BLT1 to increase [Ca]. The purpose of this study was to determine the expression of ChemR23 and BLT1 and receptors on conjunctival goblet cells and the respective roles these two receptors play in goblet cell responses to RvE1.

Methods: Goblet cells were cultured from male rat or human conjunctiva from both sexes. Western blotting analysis, reverse transcription PCR and immunofluorescence microscopy were used to demonstrate the expression of ChemR23 and BLT1 in conjunctival goblet cells. High molecular weight glycoprotein secretion was determined using an enzyme-linked lectin assay. Signaling pathways were studied by measuring the increase in [Ca] using fura 2/AM.

Results: ChemR23 and BLT1 and receptors were present on both rat and human conjunctival goblet cells. The BLT1 inhibitors LY293111 and U75302 significantly blocked RvE1-and LTB-stimulated [Ca] increase. RvE1-and LTB-stimulated [Ca] and secretion increases were blocked by BLT1-targeted siRNA. RvE1-stimulated [Ca] and secretion increases were also blocked by ChemR23-targeted siRNA. Addition of RvE1 2 min before or simultaneously with LTB desensitized the LTB [Ca] response. Addition of RvE1 and LTB simultaneously caused secretion that was decreased compared to either response alone.

Conclusion: RvE1, in addition to the ChemR23 receptor, uses the BLT1 receptor to increase [Ca] and stimulate secretion in both rat and human cultured conjunctival goblet cells.
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http://dx.doi.org/10.1016/j.jtos.2020.04.011DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7301495PMC
July 2020

Distinct Subsets of Noncoding RNAs Are Strongly Associated With BMD and Fracture, Studied in Weight-Bearing and Non-Weight-Bearing Human Bone.

J Bone Miner Res 2020 06 2;35(6):1065-1076. Epub 2020 Mar 2.

Unger-Vetlesen Institute, Lovisenberg Diaconal Hospital, Oslo, Norway.

We investigated mechanisms resulting in low bone mineral density (BMD) and susceptibility to fracture by comparing noncoding RNAs (ncRNAs) in biopsies of non-weight-bearing (NWB) iliac (n = 84) and weight bearing (WB) femoral (n = 18) postmenopausal bone across BMDs varying from normal (T-score > -1.0) to osteoporotic (T-score ≤ -2.5). Global bone ncRNA concentrations were determined by PCR and microchip analyses. Association with BMD or fracture, adjusted by age and body mass index, were calculated using linear and logistic regression and least absolute shrinkage and selection operator (Lasso) analysis. At 10% false discovery rate (FDR), 75 iliac bone ncRNAs and 94 femoral bone ncRNAs were associated with total hip BMD. Eight of the ncRNAs were common for the two sites, but five of them (miR-484, miR-328-3p, miR-27a-5p, miR-28-3p, and miR-409-3p) correlated positively to BMD in femoral bone, but negatively in iliac bone. Of predicted pathways recognized in bone metabolism, ECM-receptor interaction and proteoglycans in cancer emerged at both sites, whereas fatty acid metabolism and focal adhesion were only identified in iliac bone. Lasso analysis and cross-validations identified sets of nine bone ncRNAs correlating strongly with adjusted total hip BMD in both femoral and iliac bone. Twenty-eight iliac ncRNAs were associated with risk of fracture (FDR < 0.1). The small nucleolar RNAs, RNU44 and RNU48, have a function in stabilization of ribosomal RNAs (rRNAs), and their association with fracture and BMD suggest that aberrant processing of rRNAs may be involved in development of osteoporosis. Cis-eQTL (expressed quantitative trait loci) analysis of the iliac bone biopsies identified two loci associated with microRNAs (miRNAs), one previously identified in a heel-BMD genomewide association study (GWAS). In this comprehensive investigation of the skeletal genetic background in postmenopausal women, we identified functional bone ncRNAs associated to fracture and BMD, representing distinct subsets in WB and NWB skeletal sites. © 2020 The Authors. Journal of Bone and Mineral Research published by American Society for Bone and Mineral Research.
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http://dx.doi.org/10.1002/jbmr.3974DOI Listing
June 2020

Simple limbal epithelial transplantation: Current status and future perspectives.

Stem Cells Transl Med 2020 03 4;9(3):316-327. Epub 2019 Dec 4.

Department of Plastic and Reconstructive Surgery and Institute for Surgical Research, Oslo University Hospital, Oslo, Norway.

Damage to limbal stem cells as a result of injury or disease can lead to limbal stem cell deficiency (LSCD). This disease is characterized by decreased vision that is often painful and may progress to blindness. Clinical features include inflammation, neovascularization, and persistent cornea epithelial defects. Successful strategies for treatment involve transplantation of grafts harvested from the limbus of the alternate healthy eye, called conjunctival-limbal autograft (CLAU) and transplantation of limbal cell sheets cultured from limbal biopsies, termed cultured limbal epithelial transplantation (CLET). In 2012, Sangwan and colleagues presented simple limbal epithelial transplantation (SLET), a novel transplantation technique that combines the benefits of CLAU and CLET and avoids the challenges associated with both. In SLET a small biopsy from the limbus of the healthy eye is divided and distributed over human amniotic membrane, which is placed on the affected cornea. Outgrowth occurs from each small explant and a complete corneal epithelium is typically formed within 2 weeks. Advantages of SLET include reduced risk of iatrogenic LSCD occurring in the healthy cornea at harvest; direct transfer circumventing the need for cell culture; and the opportunity to perform biopsy harvest and transplantation in one operation. Success so far using SLET is comparable with CLAU and CLET. Of note, 336 of 404 (83%) operations using SLET resulted in restoration of the corneal epithelium, whereas visual acuity improved in 258 of the 373 (69%) reported cases. This review summarizes the results of 31 studies published on SLET since 2012. Progress, advantages, challenges, and suggestions for future studies are presented.
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http://dx.doi.org/10.1002/sctm.19-0203DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7031634PMC
March 2020

Diagnostic Test Efficacy of Meibomian Gland Morphology and Function.

Sci Rep 2019 11 22;9(1):17345. Epub 2019 Nov 22.

Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway.

Meibomian gland dysfunction (MGD) is the leading cause of dry eye and proposed treatments are based on disease severity. Our purpose was to establish reliable morphologic measurements of meibomian glands for evaluating MGD severity. This retrospective, cross-sectional study included 100 MGD patients and 20 controls. The patients were classified into dry eye severity level (DESL) 1-4 based on symptoms and clinical parameters including tear-film breakup time, ocular staining and Schirmer I. The gland loss, length, thickness, density and distortion were analyzed. We compared the morphology between patients and controls; examined their correlations to meibum expressibility, quality, and DESL. Relative to controls, the gland thickness, density and distortion were elevated in patients (p < 0.001 for all tests). The area under the receiver operating characteristic curve was 0.98 (95% confidence interval [CI], 0.96-1.0) for gland loss, and 0.96 (CI 0.91-1.0) for gland distortion, with a cutoff value of six distorted glands yielding a sensitivity of 93% and specificity of 97% for MGD diagnosis. The gland distortion was negatively correlated to the meibum expressibility (r = -0.53; p < 0.001) and DESL (r = -0.22, p = 0.018). In conclusion, evaluation of meibomian gland loss and distortion are valuable complementary clinical parameters to assess MGD status.
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http://dx.doi.org/10.1038/s41598-019-54013-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6874668PMC
November 2019

Visual Contrast Sensitivity Correlates to the Retinal Degeneration in Rhodopsin Knockout Mice.

Invest Ophthalmol Vis Sci 2019 10;60(13):4196-4204

Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States.

Purpose: Clinical manifestations of photoreceptor degeneration include gradual thinning of the outer nuclear layer (ONL) and progressive reduction of electroretinogram (ERG) amplitudes and vision loss. Although preclinical evaluations of treatment strategies greatly depend on rodent models, the courses of these changes in mice remain unclear. We thus sought to investigate the temporal correlations in changes of spatial vision, ERG response, and ONL thickness in mice with progressive photoreceptor degeneration.

Methods: Adult wild-type (WT) mice and mice carrying rhodopsin deficiency (Rho-/-), a frequently used mouse model of human retinitis pigmentosa, were selected for investigation. Mouse spatial vision, including visual acuity (VA) and contrast sensitivity (CS), was determined using optomotor response (OMR) assays; ONL thickness was quantified by spectral-domain optical coherence tomography (SD-OCT), and ERG was performed to evaluate retinal functions. The mice were killed when they were 14 weeks old, and the cone photoreceptors in retinal sections were counted.

Results: Spatial vision, ONL thickness, and ERG amplitudes remained stable in WT mice at all examined time points. While 6-week-old Rho-/- mice had VA, CS, as well as ERG responses similar to those of WT mice, progressive reductions in the spatial vision and retinal functions were recorded thereafter. Most tested 12-week-old Rho-/- mice had no visual-evoked OMR and ERG responses. Moreover, CS, but not VA, displayed a linear decline that was closely associated with ONL thinning, reduction of ERG amplitudes, and loss of cones.

Conclusions: We presented a comprehensive study of the relation between the changes of spatial vision, retinal function, and ONL thickness in postnatal week (PW)6 to PW12 Rho-/- mice. CS is a more sensitive indicator of spatial vision compared to VA, although both are required as separate parameters for monitoring the visual changes in retina undergoing photoreceptor degeneration.
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http://dx.doi.org/10.1167/iovs.19-26966DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6795341PMC
October 2019

Characteristics and Utility of Fundus Autofluorescence in Congenital Aniridia Using Scanning Laser Ophthalmoscopy.

Invest Ophthalmol Vis Sci 2019 10;60(13):4120-4128

Department of Ophthalmology, Oslo University Hospital, Oslo, Norway.

Purpose: To investigate fundus autofluorescence (FAF) and other fundus manifestations in congenital aniridia.

Methods: Fourteen patients with congenital aniridia and 14 age- and sex-matched healthy controls were examined. FAF images were obtained with an ultra-widefield scanning laser ophthalmoscope. FAF intensity was quantified in the macular fovea and in a macular ring surrounding fovea and related to an internal reference within each image. All aniridia patients underwent an ophthalmologic examination, including optical coherence tomography and slit-lamp biomicroscopy.

Results: Mean age was 28.4 ± 15.0 years in both the aniridia and control groups. Fovea could be defined by subjective assessment of FAF images in three aniridia patients (21.4%) and in all controls (P = 0.001). Mean ratio between FAF intensity in the macular ring and fovea was 1.01 ± 0.15 in aniridia versus 1.18 ± 0.09 in controls (P = 0.034). In aniridia, presence of foveal hypoplasia evaluated by biomicroscopy correlated with lack of foveal appearance by subjective analyses of FAF images (P = 0.031) and observation of nystagmus (P = 0.009).

Conclusions: Aniridia patients present a lower ratio between FAF intensity in the peripheral and central macula than do healthy individuals. Both subjective and objective analyses of FAF images are useful tools in evaluation of foveal hypoplasia in aniridia.
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http://dx.doi.org/10.1167/iovs.19-26994DOI Listing
October 2019

Functional and Morphological Evaluation of Meibomian Glands in the Assessment of Meibomian Gland Dysfunction Subtype and Severity.

Am J Ophthalmol 2020 01 14;209:160-167. Epub 2019 Sep 14.

Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway; Department of Ophthalmology, Sørlandet Hospital, Arendal, Norway; Department of Ophthalmology, Oslo University Hospital, Oslo, Norway; Institute of Clinical Dentistry/Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway; Faculty of Health Sciences, National Centre for Optics, Vision and Eye Care, University College of Southeast Norway, Kongsberg, Norway; Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Oslo, Norway; Department of Ophthalmology, Stavanger University Hospital, Stavanger, Norway.

Purpose: To classify subtypes of meibomian gland dysfunction (MGD) and evaluate the dependency of dry eye signs, symptoms, and parameters on MGD subtype.

Design: Cross-sectional study.

Study Population: the right eyes of 447 patients with MGD of various subtypes and 20 healthy volunteers.

Methods: Patients were divided into 4 subtypes of MGD based on meibum expression, meibum quality, and MG loss on meibography images (meibograde of 0-6). Subtypes were patients with high meibum delivery (hypersecretory and nonobvious MGD) and those with low meibum delivery (hyposecretory and obstructive MGD). Additional clinical tests included tear film break-up time (TFBUT), ocular staining, osmolarity, Schirmer I, blink interval timing and the Ocular Surface Disease Index (OSDI) questionnaire.

Results: A total of 78 eyes had hypersecretory MGD; 49 eyes had nonobvious MGD; 66 eyes had hyposecretory MGD; and 254 eyes had obstructive MGD. Increased tear film osmolarity and lower TFBUT were found in the low-delivery groups; hyposecretory (P = 0.006, P = 0.016) and obstructive MGD (P = 0.008, P = 0.006) relative to high-delivery MGD (hypersecretory and nonobvious groups, respectively). Worse ocular symptoms and ocular staining were also found in low-delivery MGD groups than the high delivery MGD groups (P < 0.01 and P < 0.006, respectively).

Conclusions: Patients with low-delivery MGD had worse dry eye parameters and ocular symptoms than those with high meibum delivery, indicating the pivotal role of meibum secretion in ocular surface health that should be targeted in MGD therapy. Furthermore, nonobvious MGD cannot be diagnosed using conventional dry eye tests and requires morphologic assessment of meibography images to confirm MG loss.
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http://dx.doi.org/10.1016/j.ajo.2019.09.005DOI Listing
January 2020

Resolvin D2 elevates cAMP to increase intracellular [Ca] and stimulate secretion from conjunctival goblet cells.

FASEB J 2019 07 23;33(7):8468-8478. Epub 2019 Apr 23.

Schepens Eye Research Institute, Massachusetts Eye and Ear-Harvard Medical School, Boston, Massachusetts, USA.

Under physiologic conditions, conjunctival goblet cells (CGCs) secrete mucins into the tear film to preserve ocular surface homeostasis. Specialized proresolving mediators (SPMs), like resolvins (Rvs), regulate secretion from CGCs and actively terminate inflammation. The purpose of this study was to determine if RvD2 stimulated mucin secretion and to investigate the cellular signaling components. Goblet cells were cultured from rat conjunctiva. Secretion was measured by an enzyme-linked lectin assay, change in intracellular [Ca] ([Ca]) using Fura-2, and cellular cAMP levels by ELISA. RvD2 (10-10 M) stimulated secretion, increased cellular cAMP levels and the [Ca]. RvD2-stimulated increase in [Ca] and secretion was blocked by Ca chelator 1,2-bis(2-aminophenoxy)ethane-,,','-tetraacetic acid tetrakis and the PKA inhibitor -[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride but not by the cAMP exchange protein inhibitor α-[2-(3-chlorophenyl)hydrazinylidene]-5-(1,1-dimethylethyl)-b-oxo-3-isoxazolepropanenitrile. Forskolin, 3-isobutyl-1-methylxanthine, and 8-bromo-cAMP (8-Br-cAMP) increased [Ca]. Increasing cAMP with 8-Br-cAMP inhibited the increase in [Ca] stimulated by the cAMP-independent agonist cholinergic agonist carbachol. In conclusion, RvD2 uses both cellular cAMP and [Ca] to stimulate glycoconjugate secretion from CGCs, but the interaction of cAMP and [Ca] is context dependent. Thus RvD2 likely assists in the maintenance of the mucous layer of the tear film to sustain ocular surface homeostasis and has potential as a novel treatment for dry eye disease.-Botten, N., Hodges, R. R., Li, D., Bair, J. A., Shatos, M. A., Utheim, T. P., Serhan, C. N., Dartt, D. A. Resolvin D2 elevates cAMP to increase intracellular [Ca] and stimulate secretion from conjunctival goblet cells.
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http://dx.doi.org/10.1096/fj.201802467RDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6593886PMC
July 2019

Evaluation of the Ocular Surface Disease Index Questionnaire as a Discriminative Test for Clinical Findings in Dry Eye Disease Patients.

Curr Eye Res 2019 09 29;44(9):941-947. Epub 2019 Apr 29.

Department of Medical Biochemistry, Oslo University Hospital , Oslo , Norway.

: To investigate to what extent the OSDI can be utilized as a discriminative test for clinical findings. : One thousand and ninety patients with dry eye disease (DED) were consecutively included and examined for osmolarity, tear film break-up time (TFBUT), ocular protection index (OPI), ocular surface staining (OSS), Schirmer I test (ST), meibum expressibility (ME), meibum quality (MQ), and diagnosis of meibomian gland dysfunction (MGD). Receiver-operating characteristic curve (ROC) analysis considering optimum balanced sensitivity and specificity (close to 50%) was used for assessment. : The present study on more than 1,000 patients indicates that the OSDI in the ROC curve analysis is a poor discriminator of pathological scores for TFBUT ≤ 5 (AUC = 0.553; = .012) and ≤10 s (AUC = 0.608; = .002), OSS ≥ 3 (AUC = 0.54; = .043), ST ≤ 5 (AUC = 0.550; = .032) and ≤10 mm/5 min (AUC = 0.544; = .016), and ME ≥ 1 (AUC = 0.594; = <0.001). Pathological scores for osmolarity >308 and >316 mOsm/L, OPI, OSS > 1, MQ, and MGD could not be discriminated by OSDI ( > .05). : Cut-off values for the OSDI can be defined to discriminate pathological TFBUT (≤5 and ≤10), OSS (≥3), ST (≤5 and ≤10) and ME, however, the discriminability was low. Our comprehensive study emphasises the importance of taking both symptoms and signs into account in DED management.
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http://dx.doi.org/10.1080/02713683.2019.1604972DOI Listing
September 2019

Conjunctival Goblet Cells, the Overlooked Cells in Glaucoma Treatment.

J Glaucoma 2019 04;28(4):325-333

Department of Drug Design and Pharmacology, University of Copenhagen.

Glaucoma is the leading cause of irreversible blindness worldwide. Although no definitive cure exists, lowering of the intraocular pressure decreases the rate of progression in the majority of patients with glaucoma. Antiglaucomatous treatment modalities consist predominantly of chronic use of eye drops. It has become increasingly evident that long-term exposure to eye drops has a significant impact on the ocular surface, and thereby on patient compliance and quality of life. Maintenance of the ocular surface is highly dependent on a stable tear film. Conjunctival goblet cells (GCs) of the ocular surface play an important role in providing the innermost mucin layer of the tear film and are essential for maintaining the ocular surface homeostasis. Recent studies have reported severe side effects of antiglaucomatous drops on GCs. In particular, a preservative containing antiglaucomatous drops have been shown to affect the viability and functions of the GCs. Furthermore, GC density has been suggested as a potential predictor of surgical outcome after filtration surgery. The present review provides an overview of the current literature on the impact of antiglaucomatous eye drops on GCs as well as the impact on the ocular surface. Moreover, the existing evidence of a possible association between GC density and glaucoma filtration surgery outcome is summarized. We conclude that prostaglandin analogs spare the conjunctival GCs more compared with other antiglaucomatous drops and that GCs may be a good predictor of surgical outcome after filtration surgery. Overall, given the multiple functions of GCs in the ocular surface homeostasis, dedicated strategies should be adopted to preserve this cell population during the course of glaucoma.
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http://dx.doi.org/10.1097/IJG.0000000000001168DOI Listing
April 2019

Meibomian Gland Morphology Is a Sensitive Early Indicator of Meibomian Gland Dysfunction.

Am J Ophthalmol 2019 04 20;200:16-25. Epub 2018 Dec 20.

Department of Oral Surgery and Oral Medicine, Faculty of Dentistry, University of Oslo, Oslo, Norway; Department of Ophthalmology, Vestre Viken Hospital Trust, Drammen, Norway; Faculty of Health Sciences, National Centre for Optics, Vision and Eye Care, University College of Southeast Norway, Kongsberg, Norway; Department of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden; Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway; Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Oslo, Norway; Department of Maxillofacial Surgery, Oslo University Hospital, Oslo, Norway.

Purpose: To investigate the relationship between meibomian gland (MG) morphology and clinical dry eye tests in patients with meibomian gland dysfunction (MGD).

Design: Cross-sectional study.

Subjects: Total 538 MGD patients and 21 healthy controls.

Methods: MG loss on meibography images of upper (UL) and lower lids (LL) was graded on a scale of 0 (lowest degree of MG loss) to 3. MG length, thickness, and interglandular space in the UL were measured. Clinical tests included meibum expression and quality, tear film break-up time, ocular staining, osmolarity, Schirmer I, blink interval timing, and Ocular Surface Disease Index (OSDI) questionnaire.

Results: Mean UL and LL meibogrades were significantly higher in MGD patients compared to controls (P < .001 for UL and LL). The sensitivity and specificity of the meibograde as a diagnostic parameter for MGD was 96.7% and 85%, respectively. Schirmer I was significantly increased in MGD patients with meibograde 1 compared to patients with meibograde 0, 2, and 3 in the UL (P < .05). MG thickness increased with higher meibograde (P < .001). MG morphology correlated significantly but weakly with several clinical parameters (P < .05). OSDI did not correlate with any MG morphologic parameter.

Conclusions: Grading of MG loss using meibograde effectively diagnoses MGD. Compensatory mechanisms such as increased aqueous tear production and dilation of MGs make early detection of MGD difficult by standard clinical measures of dry eye, whereas morphologic analysis of MGs reveals an early stage of MGD, and therefore represents a complementary clinical parameter with diagnostic potential.
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http://dx.doi.org/10.1016/j.ajo.2018.12.006DOI Listing
April 2019

Resolvin D1, but not resolvin E1, transactivates the epidermal growth factor receptor to increase intracellular calcium and glycoconjugate secretion in rat and human conjunctival goblet cells.

Exp Eye Res 2019 03 1;180:53-62. Epub 2018 Dec 1.

Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, USA. Electronic address:

Purpose: To identify interactions of the epidermal growth factor receptor (EGFR) with the pro-resolving mediator receptors for RvD1 and RvE1 to stimulate an increase in intracellular [Ca] ([Ca]) and mucin secretion from cultured human and rat conjunctival goblet cells.

Methods: Goblet cells from human and rat conjunctiva were grown in culture using RPMI media. Cultured goblet cells were pre-incubated with inhibitors, and then stimulated with RvD1, RvE1, EGF or the cholinergic agonist carbachol (Cch). Increase in [Ca] was measured using fura-2/AM. Goblet cell secretion was measured using an enzyme-linked lectin assay with UEA-1. Western blot analysis was performed with antibodies against AKT and ERK 1/2.

Results: In cultured human conjunctival goblet cells RvE1 -stimulated an increase in [Ca]. RvD1-, but not the RvE1-, stimulated increase in [Ca] and mucin secretion was blocked by the EGFR inhibitor AG1478 and siRNA for the EGFR. RvD1-, but not RvE1-stimulated an increase in [Ca] that was also inhibited by TAPI-1, an inhibitor of the matrix metalloprotease ADAM 17. Inhibition of the EGFR also blocked RvD1-stimulated increase in AKT activity and both RvD1-and RvE1-stimulated increase in ERK 1/2 activity. Pretreatment with either RvD1 or RvE1 did not block the EGFR-stimulated increase in [Ca].

Conclusions: We conclude that in cultured rat and human conjunctival goblet cells, RvD1 activates the EGFR, increases [Ca], activates AKT and ERK1/2 to stimulate mucin secretion. RvE1 does not transactivate the EGFR to increase [Ca] and stimulate mucin secretion, but does interact with the receptor to increase ERK 1/2 activity.
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http://dx.doi.org/10.1016/j.exer.2018.11.018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6389367PMC
March 2019

Coordinated collective migration and asymmetric cell division in confluent human keratinocytes without wounding.

Nat Commun 2018 09 10;9(1):3665. Epub 2018 Sep 10.

Department of Medical Biochemistry, Oslo University Hospital, 0372, Oslo, Norway.

Epithelial sheet spreading is a fundamental cellular process that must be coordinated with cell division and differentiation to restore tissue integrity. Here we use consecutive serum deprivation and re-stimulation to reconstruct biphasic collective migration and proliferation in cultured sheets of human keratinocytes. In this system, a burst of long-range coordinated locomotion is rapidly generated throughout the cell sheet in the absence of wound edges. Migrating cohorts reach correlation lengths of several millimeters and display dependencies on epidermal growth factor receptor-mediated signaling, self-propelled polarized migration, and a G/G cell cycle environment. The migration phase is temporally and spatially aligned with polarized cell divisions characterized by pre-mitotic nuclear migration to the cell front and asymmetric partitioning of nuclear promyelocytic leukemia bodies and lysosomes to opposite daughter cells. This study investigates underlying mechanisms contributing to the stark contrast between cells in a static quiescent state compared to the long-range coordinated collective migration seen in contact with blood serum.
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http://dx.doi.org/10.1038/s41467-018-05578-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6131553PMC
September 2018

Tear Production Levels and Dry Eye Disease Severity in a Large Norwegian Cohort.

Curr Eye Res 2018 12 5;43(12):1465-1470. Epub 2018 Sep 5.

a Department of Medical Biochemistry , Oslo University Hospital , Oslo , Norway.

Purpose: To determine if the Schirmer I test (without anesthesia) cut-off value is a predictor of dry eye severity in a large Norwegian cohort of dry eye disease (DED) patients, which are grouped into six levels of tear production.

Methods: Patients (n = 1090) with DED of different etiologies received an extensive dry eye work-up: osmolarity (Osm), tear meniscus height (TMH), tear film break-up time (TFBUT), ocular protection index (OPI), ocular surface staining (OSS), Schirmer I test (ST), meibum expressibility (ME), and meibum quality (MQ). Classification of dry eye severity level (DESL) and diagnosis of meibomian gland dysfunction (MGD) were also included. The cohort was divided into six groups: below and above cut-off values of 5 (groups 1 and 2), 10 (groups 3 and 4), and 15 mm (groups 5 and 6) of ST. Mann-Whitney test and Chi-Square test were used for group comparison of parameters (p ≤ 0.05).

Results: The groups 1, 3, and 5 had values indicating more severe DED than the groups 2, 4, 6 with significant difference in DESL, Osm, TFBUT, OPI, OSS, and TMH. Regardless of the choice of cut-off values, there was no statistically significant difference in ME, MQ, and MGD between groups below and above selected cut-off value. When gender difference was considered in each group, significant difference was only observed for DESL (groups 2, 4, and 5), TFBUT (groups 2, 4, and 5), OPI (groups 2 and 6), and ME (group1).

Conclusions: Schirmer I is a robust discriminator for DESL, Osm, TFBUT, OPI, OSS, and TMH, but not for ME, MQ, and MGD. Patients with lower tear production levels presented with more severe DED at all three defined cut-off values. Interestingly, the differences in the mean values of DESL were minimal although statistically significant. Thus, the clinical value of different Schirmer levels appears to be limited.
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http://dx.doi.org/10.1080/02713683.2018.1514055DOI Listing
December 2018

Activation of the EGF Receptor by Histamine Receptor Subtypes Stimulates Mucin Secretion in Conjunctival Goblet Cells.

Invest Ophthalmol Vis Sci 2018 07;59(8):3543-3553

Schepens Eye Research Institute, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States.

Purpose: The purpose of this study was to determine if histamine receptors interact with the epidermal growth factor receptor (EGFR) in cultured rat conjunctival goblet cells.

Methods: Goblet cells from rat conjunctiva were grown in organ culture. First-passage goblet cells were used in all experiments. Phosphorylated (active) and total EGFR, AKT, and extracellular signal-regulated kinase (ERK)1/2 were measured by Western blot analysis. Cells were preincubated with the EGFR antagonist AG1478 for 30 minutes or small interfering RNA specific to the EGFR for 3 days prior to stimulation with histamine or agonists specific for histamine receptor subtypes for 2 hours. Goblet cell secretion was measured using an enzyme-linked lectin assay. Goblet cells were incubated for 1 hour with the calcium indicator molecule fura-2/AM, and intracellular [Ca2+] ([Ca2+]i) was determined. Data were collected in real time and presented as the actual [Ca2+]i with time and as the change in peak [Ca2+]i.

Results: Histamine increased the phosphorylation of the EGFR. Mucin secretion and increase in [Ca2+]i stimulated by histamine, and agonists specific for each histamine receptor subtype were blocked by inhibition of the EGFR. Increase in [Ca2+]i stimulated by histamine and specific agonists for each histamine receptor was also inhibited by TAPI-1, a matrix metalloproteinase (MMP) inhibitor. The histamine-stimulated increase in activation of AKT, but not ERK1/2, was blocked by AG1478.

Conclusions: In conjunctival goblet cells, histamine, using all four receptor subtypes, transactivates the EGFR via an MMP. This in turn phosphorylates AKT to increase [Ca2+]i and stimulate mucin secretion.
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http://dx.doi.org/10.1167/iovs.18-2476DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6049985PMC
July 2018

The Level of Inflammatory Tear Cytokines is Elevated in Congenital Aniridia and Associated with Meibomian Gland Dysfunction.

Invest Ophthalmol Vis Sci 2018 04;59(5):2197-2204

Department of Ophthalmology, Oslo University Hospital, Oslo, Norway.

Purpose: To investigate the tear cytokine profile in congenital aniridia, and correlate cytokine levels with ophthalmologic findings.

Methods: We examined 35 patients with aniridia and 21 healthy controls. Tear fluid was collected with Schirmer I test and capillary tubes from each eye, and the concentration of 27 inflammatory cytokines determined using multiplex bead assay. Eyes of all participants were examined with tests for dry eye disease, including evaluation of meibomian glands (meibography). Differences in cytokine levels between the two groups were analyzed, and correlations between cytokine concentrations and ophthalmologic findings in the aniridia group investigated.

Results: The concentrations of six tear cytokines were significantly higher in aniridia patients than controls in both eyes, and included interleukin 1β (IL-1β), IL-9, IL-17A; eotaxin; basic fibroblast growth factor (bFGF/FGF2); and macrophage inflammatory protein 1α (MIP-1α/CCL3). The ratio between the anti-inflammatory IL-1RA and the proinflammatory IL-1β was significantly lower in patients than controls in both eyes (P = 0.005 right eye and P = 0.001 left eye). Increasing concentration of IL-1β, IL-9, IL-17A, FGF2, and MIP-1α correlated with parameters for meibomian gland dysfunction (MGD) in the aniridia group, including increasing atrophy of meibomian glands, and shorter break-up time of the tear film.

Conclusions: A number of pro-inflammatory cytokines are significantly elevated in tear fluid from aniridia patients, and correlate with parameters for MGD in aniridia. Increased inflammation of the ocular surface may be a factor in the development of MGD in aniridia patients, and explain the high prevalence of MGD and dry eye disease in these patients.
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http://dx.doi.org/10.1167/iovs.18-24027DOI Listing
April 2018