1. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7496-7515. doi:
10.1167/iovs.15-17616.

The bHLH Transcription Factor NeuroD Governs Photoreceptor Genesis and
Regeneration Through Delta-Notch Signaling.

Taylor SM(1), Alvarez-Delfin K(2), Saade CJ(2), Thomas JL(3), Thummel R(3),
Fadool JM(2), Hitchcock PF(1).

Author information: 
(1)Department of Ophthalmology and Visual Sciences University of Michigan, W. K. 
Kellogg Eye Center, Ann Arbor, Michigan, United States. (2)Department of
Biological Science, Florida State University, Tallahassee, Florida, United
States. (3)Departments of Anatomy/Cell Biology and Ophthalmology, Wayne State
University, Detroit, Michigan, United States.

Purpose: Photoreceptor genesis in the retina requires precise regulation of
progenitor cell competence, cell cycle exit, and differentiation, although
information around the mechanisms that govern these events currently is lacking. 
In zebrafish, the basic helix-loop-helix (bHLH) transcription factor NeuroD
governs photoreceptor genesis, but the signaling pathways through which NeuroD
functions are unknown. The purpose of this study was to identify these pathways, 
and during photoreceptor genesis, Notch signaling was investigated as the
putative mediator of NeuroD function.
Methods: In embryos, genetic mosaic analysis was used to determine if NeuroD
functions is cell- or non-cell-autonomous. Morpholino-induced NeuroD knockdown,
CRISPR/Cas9 mutation, and pharmacologic and transgenic approaches were used,
followed by in situ hybridization, immunocytochemistry, and quantitative RT-PCR
(qRT-PCR), to identify mechanisms through which NeuroD functions. In adults,
following photoreceptor ablation and NeuroD knockdown, similar methods as above
were used to identify NeuroD function during photoreceptor regeneration.
Results: In embryos, NeuroD function is non-cell-autonomous, NeuroD knockdown
increases Notch pathway gene expression, Notch inhibition rescues the NeuroD
knockdown-induced deficiency in cell cycle exit but not photoreceptor maturation,
and Notch activation and CRISPR/Cas9 mutation of neurod recapitulate NeuroD
knockdown. In adults, NeuroD knockdown prevents cell cycle exit and photoreceptor
regeneration and increases Notch pathway gene expression, and Notch inhibition
rescues this phenotype.
Conclusions: These data demonstrate that during embryonic development, NeuroD
governs photoreceptor genesis via non-cell-autonomous mechanisms and that, during
photoreceptor development and regeneration, Notch signaling is a mechanistic link
between NeuroD and cell cycle exit. In contrast, during embryonic development,
NeuroD governs photoreceptor maturation via mechanisms that are independent of
Notch signaling.

PMID: 26580854   [PubMed - as supplied by publisher]


2. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7438-7443. doi:
10.1167/iovs.15-17449.

Retinal Vascular Fractals Correlate With Early Neurodegeneration in Patients With
Type 2 Diabetes Mellitus.

Frydkjaer-Olsen U(1), Soegaard Hansen R(1), Pedersen K(1), Peto T(2), Grauslund
J(1).

Author information: 
(1)Department of Ophthalmology, Odense University Hospital, Odense, Denmark
2Institute of Clinical Research, University of Southern Denmark, Odense, Denmark.
(2)Institute of Clinical Research, University of Southern Denmark, Odense,
Denmark 3The NIHR Biomedical Research Centre, Moorfields Eye Hospital NHS
Foundation Trust and UCL Institute of Ophthalmology, London, United Kingdom.

Purpose: To investigate the correlation between the retinal vascular fractal
dimension (Fd) and neurodegenerative changes in patients with no or mild diabetic
retinopathy (DR).
Methods: In this cross-sectional study we examined 103 patients with type 2
diabetes mellitus (T2DM) with no or mild DR. In a randomly selected eye of each
patient, Fd was calculated using SIVA-Fractal, a specialized semiautomatic
software. Retinal neurodegeneration was evaluated by Topcon 3D OCT-2000
spectral-domain optical coherence tomography (OCT) and by a RETI-scan multifocal 
ERG (mf-ERG) system in rings one to six. Level of DR was determined by a single
trained grader in seven-field fundus photos according to the Early Treatment
Diabetic Retinopathy Study (ETDRS) scale.
Results: Mean age and duration of T2DM were 62.3 and 11.6 years, respectively;
46.6% were men. Mean Fd was 1.413 (range, 1.278-1.509) and ETDRS levels were 10
(42.7%), 20 (35.0%), and 35 (22.3%), respectively. Fd correlated inversely with
mf-ERG implicit time of ring one (r = -0.25, P = 0.01) and present diabetic
neuropathy (P = 0.02), and positively with OCT ganglion cell layer (GCL)
thickness (r = 0.20, P = 0.04). In a multivariable linear regression model, Fd
was associated with mf-ERG implicit time of ring one (coefficient -0.0021/ms, P =
0.040) and the presence of diabetic neuropathy (coefficient -0.0209 for
neuropathy present versus absent, P = 0.041).
Conclusions: In patients with T2DM and no or minimal DR, independent correlations
were found between early vascular and neurogenic changes. Thus, retinal vascular 
fractal analysis might be considered as a tool to identify patients with early
neurodegenerative retinal changes.

PMID: 26580853   [PubMed - as supplied by publisher]


3. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7427-7437. doi:
10.1167/iovs.15-17705.

Visual Pathways in Humans With Ephrin-B1 Deficiency Associated With the
Cranio-Fronto-Nasal Syndrome.

Hoffmann MB(1), Thieme H(2), Liedecke K(2), Meltendorf S(2), Zenker M(3), Wieland
I(3).

Author information: 
(1)Department of Ophthalmology Otto-von-Guericke-University, Magdeburg, Germany
2Center for Behavioural Brain Sciences, Magdeburg, Germany. (2)Department of
Ophthalmology Otto-von-Guericke-University, Magdeburg, Germany. (3)Institute for 
Human Genetics, Otto-von-Guericke-University, Magdeburg, Germany.

Purpose: Numerous animal studies demonstrated the importance of components of the
ephrin/Eph system for correct visual system development. Analogous investigations
in humans are entirely missing. Here, we examined the visual system in humans
with ephrin-B1 deficiency, which is x-linked and associated with the
cranio-fronto-nasal syndrome (CFNS) in heterozygous females.
Methods: For one male hemizygous for ephrin-B1 deficiency and three affected
heterozygous females with molecular-genetically confirmed mutations, the
integrity of the partial decussation of the optic nerves was assessed with visual
evoked potentials (VEPs) and compared with albinotic, achiasmic, and control
participants with healthy vision. Further, retinal morphology and function and
the gross-retinotopic representation of the primary visual cortex were examined
with spectral-domain optical coherence tomography (SD-OCT), ERG, and multifocal
(mf) VEPs for the male participant and part of the carriers.
Results: Strabismus and lack of stereovision was evident in the male and two of
the females. Other characteristics of the visual system organization and function
were normal: (1) retina: SD-OCT and funduscopy indicated normal foveal and optic 
nerve head morphology. Electroretinograms indicated normal retinal function, (2) 
optic chiasm: conventional (c)VEP showed no evidence for misrouting and mfVEPs
were only suggestive of, if any, very minor local misrouting, and (3) visual
cortex: mfVEP characteristics indicated normal retinotopic gross-representations 
of the contralateral visual hemifield in each hemisphere.
Conclusions: While ephrin-B1 deficiency leads to abnormal visual pathways in
mice, it leaves the human visual system, apart from deficits in binocular vision,
largely normal. We presume that other components of the ephrin-system can
substitute the lack of ephrin-B1 in humans.

PMID: 26580852   [PubMed - as supplied by publisher]


4. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7418-7426. doi:
10.1167/iovs.15-17920.

A Nonsense Mutation in FAM161A Is a Recurrent Founder Allele in Dutch and Belgian
Individuals With Autosomal Recessive Retinitis Pigmentosa.

Van Schil K(1), Klevering BJ(2), Leroy BP(3), Pott JW(4), Bandah-Rozenfeld D(5), 
Zonneveld-Vrieling MN(6), Sharon D(5), den Hollander AI(7), Cremers FP(8), De
Baere E(1), Collin RW(8), van den Born LI(9).

Author information: 
(1)Center for Medical Genetics Ghent University and Ghent University Hospital,
Ghent, Belgium. (2)Department of Ophthalmology, Radboud University Medical
Center, Nijmegen, The Netherlands 3Radboud Institute for Health Sciences, Radboud
University Medical Center, Nijmegen, The Netherlands. (3)Center for Medical
Genetics Ghent University and Ghent University Hospital, Ghent, Belgium
4Department of Ophthalmology, Ghent University Hospital, Ghent, Belgium 5Division
of Ophthalmology, Children's Hospital of Philadelphia, Philadelphia,
Pennsylvania. (4)Department of Ophthalmology, University of Groningen, University
Medical Centre Groningen, Groningen, The Netherlands. (5)Department of
Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
(6)Department of Human Genetics, Radboud University Medical Center, Nijmegen, The
Netherlands. (7)Department of Ophthalmology, Radboud University Medical Center,
Nijmegen, The Netherlands 8Department of Human Genetics, Radboud University
Medical Center, Nijmegen, The Netherlands 9Radboud Institute for Molecular Life
Sciences, Radboud University Medica. (8)Department of Human Genetics, Radboud
University Medical Center, Nijmegen, The Netherlands 9Radboud Institute for
Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The
Netherlands. (9)The Rotterdam Eye Hospital, Rotterdam, The Netherlands.

Purpose: To identify mutations in FAM161A underlying autosomal recessive
retinitis pigmentosa (arRP) in the Dutch and Belgian populations and to
investigate whether common FAM161A-associated phenotypic features could be
identified.
Methods: Homozygosity mapping, amplification-refractory mutation system (ARMS)
analysis, and Sanger sequencing were performed to identify mutations in FAM161A. 
Microsatellite and SNP markers were genotyped for haplotype analysis. Patients
with biallelic mutations underwent detailed ophthalmologic examinations,
including measuring best-corrected visual acuity, extensive fundus photography
with reflectance and autofluorescence imaging, and optical coherence tomography.
Results: Homozygosity mapping in 230 Dutch individuals with suspected arRP
yielded five individuals with a homozygous region harboring FAM161A. Sanger
sequencing revealed a homozygous nonsense mutation (c.1309A>T; p.[Arg437*]) in
one individual. Subsequent ARMS analysis and Sanger sequencing in Dutch and
Belgian arRP patients resulted in the identification of seven additional
individuals carrying the p.(Arg437*) mutation, either homozygously or compound
heterozygously with another mutation. Haplotype analysis identified a shared
haplotype block of 409 kb surrounding the p.(Arg437*) mutation in all patients,
suggesting a founder effect. Although the age of onset was variable among
patients, all eight developed pronounced outer retinal loss with severe visual
field defects and a bull's eye-like maculopathy, followed by loss of central
vision within 2 decades after the initial diagnosis in five subjects.
Conclusions: A founder mutation in FAM161A p.(Arg437*) underlies approximately 2%
of arRP cases in the Dutch and Belgian populations. The age of onset of the
retinal dystrophy appears variable, but progression can be steep, with almost
complete loss of central vision later in life.

PMID: 26574802   [PubMed - as supplied by publisher]


5. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7417. doi: 10.1167/iovs.15-18531.

Adult Human Choroid: An Alymphatic Tissue?

Lutty GA.

PMID: 26574801   [PubMed - as supplied by publisher]


6. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7406-7416. doi:
10.1167/iovs.15-17883.

Lymphatic Markers in the Adult Human Choroid.

Schrödl F(1), Kaser-Eichberger A(2), Trost A(2), Strohmaier C(2), Bogner B(2),
Runge C(2), Motloch K(2), Bruckner D(2), Laimer M(3), Heindl LM(4), Reitsamer
HA(2).

Author information: 
(1)University Clinic of Ophthalmology and Optometry Research Program for
Experimental Ophthalmology and Glaucoma Research, Paracelsus Medical University, 
SALK, Salzburg, Austria 2Department of Anatomy, Paracelsus Medical University,
Salzburg, Austria. (2)University Clinic of Ophthalmology and Optometry Research
Program for Experimental Ophthalmology and Glaucoma Research, Paracelsus Medical 
University, SALK, Salzburg, Austria. (3)University Clinic of Dermatology,
Paracelsus Medical University, SALK, Salzburg, Austria. (4)Department of
Ophthalmology, University of Cologne, Cologne, Germany.

Purpose: Reports of lymphatics in the posterior human uvea are contradictory. We 
systematically analyzed the choroid by combining various lymphatic markers,
following recently established guidelines for the immunohistochemical detection
of ocular lymphatics.
Methods: Human choroids were prepared for flat mount serial cryosectioning.
Sections were processed for immunohistochemistry of the lymphatic markers LYVE-1,
PDPN, PROX1, FOXC2, VEGFR3, CCL21, and combined with α-smooth muscle-actin and
4',6-diamidino-2-phenylendole (DAPI). Single, double, and triple marker
combinations were documented using confocal microscopy. Messenger RNA analysis
for CCL21, FOXC2, LYVE-1, PDPN, PROX, and VEGFR3 was performed in choroid and
skin.
Results: In the choroid, CCL21 immunoreactivity was detected in choroidal blood
vessels, intrinsic choroidal neurons, and numerous small cells of the choroidal
stroma. These small cells were not colocalized with PROX1 and PDPN, while a
subpopulation of cells showed immunoreactivity for CCL21 and LYVE-1, and very
occasionally PDPN-only+ cells were detected. Nuclei positive for PROX1 were never
detected in the choroid, and vessel-like structures immunoreactive for LYVE-1,
PDPN, or CCL21 (other than blood vessels) were never observed. Immunoreactivity
of VEGFR3 was absent in the majority of choroidal blood vessels, but present in
choriocapillaris, while other structures positive for VEGFR3 were not detected.
Nonvascular smooth muscle cells were lacking VEGFR3-immunoreactivity. Messenger
RNA analysis detected all lymphatic markers investigated and confirmed
immunohistochemical results.
Conclusions: By combining several lymphatic markers, single cells expressed these
markers, but classical lymphatic vessels were not detected in the human choroid. 
Therefore, the healthy adult human choroid must be considered alymphatic, at
least with the markers applied here.

PMID: 26574800   [PubMed - as supplied by publisher]


7. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7398-7405. doi:
10.1167/iovs.15-18117.

MALDI Imaging Mass Spectrometry Spatially Maps Age-Related Deamidation and
Truncation of Human Lens Aquaporin-0.

Wenke JL, Rose KL, Spraggins JM, Schey KL.

Purpose: To spatially map human lens Aquaporin-0 (AQP0) protein modifications,
including lipidation, truncation, and deamidation, from birth through middle age 
using matrix-assisted laser desorption ionization (MALDI) imaging mass
spectrometry (IMS).
Methods: Human lens sections were water-washed to facilitate detection of
membrane protein AQP0. We acquired MALDI images from eight human lenses ranging
in age from 2 months to 63 years. In situ tryptic digestion was used to generate 
peptides of AQP0 and peptide images were acquired on a 15T Fourier transform ion 
cyclotron resonance (FTICR) mass spectrometer. Peptide extracts were analyzed by 
liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database searched
to identify peptides observed in MALDI imaging experiments.
Results: Unmodified, truncated, and fatty acid-acylated forms of AQP0 were
detected in protein imaging experiments. Full-length AQP0 was fatty acid acylated
in the core and cortex of young (2- and 4-month) lenses. Acylated and unmodified 
AQP0 were C-terminally truncated in older lens cores. Deamidated tryptic peptides
(+0.9847 Da) were mass resolved from unmodified peptides by FTICR MS. Peptide
images revealed differential localization of un-, singly-, and doubly-deamidated 
AQP0 C-terminal peptide (239-263). Deamidation was present at 4 months and
increases with age. Liquid chromatography-MS/MS results indicated N246 undergoes 
deamidation more rapidly than N259.
Conclusions: Results indicated AQP0 fatty acid acylation and deamidation occur
during early development. Progressive age-related AQP0 processing, including
deamidation and truncation, was mapped in human lenses as a function of age. The 
localization of these modified AQP0 forms suggests where AQP0 functions may
change throughout lens development and aging.

PMID: 26574799   [PubMed - as supplied by publisher]


8. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7388-7397. doi:
10.1167/iovs.15-16899.

Cerebral Involvement in Stargardt's Disease: A VBM and TBSS Study.

Gaia O(1), Melillo P(2), Sirio C(1), D'Alterio FM(2), Prinster A(3), Testa F(2), 
Brunetti A(1), Simonelli F(2), Quarantelli M(3).

Author information: 
(1)Department of Advanced Biomedical Sciences, University "Federico II," Naples, 
Italy. (2)Multidisciplinary Department of Medical, Surgical, and Dental Sciences,
Eye Clinic, Second University of Naples, Naples, Italy. (3)Institute of
Biostructure and Bioimaging, National Research Council, Naples, Italy.

Purpose: To assess whether and to what extent macro- and/or microstructural
modifications are present in the brain of patients with selective central visual 
loss due to a juvenile macular degeneration, Stargardt's disease (STGD), taking
advantage of the complementary information provided by voxel-based morphometry
(VBM) and diffusion tensor imaging (DTI).
Methods: Eighteen patients with clinical and molecular diagnosis of STGD related 
to ABCA4 mutations and 23 normally sighted volunteers of comparable age and sex
were enrolled. Structural T1-weighted (T1w) volumes, for brain tissue volume
assessment by segmentation, and DTI, for the investigation of diffusivity
parameters via a tract-based spatial statistics (TBSS) procedure, were acquired
at 3 Tesla in all subjects. All patients underwent a complete ophthalmologic
examination, including best-corrected visual acuity (BCVA), biomicroscopy,
ophthalmoscopy, electroretinography (ERG), microperimetry, and optical coherence 
tomography (OCT). Correlations between imaging data and clinical measures were
tested.
Results: Stargardt's disease patients showed a significant gray matter (GM) loss 
bilaterally in the occipital cortices, extending into the right precuneus, and in
the fronto-orbital cortices. At TBSS, significant reductions in fractional
anisotropy were detected throughout large regions in the supratentorial white
matter (WM), more pronounced in the posterior areas. Gray matter volume
correlated directly with mean visual sensitivity in the right middle frontal and 
left calcarine gyri, and inversely with retinal thickness in the left
supramarginal gyrus.
Conclusions: In STGD, widespread microstructural WM alterations are present,
suggestive of minor fiber loss coupled with GM loss, also in cortical regions not
traditionally linked to visual pathways, at least partly related to the retinal
damage.

PMID: 26574798   [PubMed - as supplied by publisher]


9. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7387. doi: 10.1167/iovs.15-18268.

Intracellular Toll-Like Receptors Help Retinal Microglia Sense Corneal
Infections.

Langmann T.

PMID: 26574797   [PubMed - as supplied by publisher]


10. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7377-7386. doi:
10.1167/iovs.15-17587.

Retinal Microglial Activation Following Topical Application of Intracellular
Toll-Like Receptor Ligands.

Chinnery HR(1), NaranjoGolborne C(2), Leong CM(2), Chen W(3), Forrester JV(4),
McMenamin PG(2).

Author information: 
(1)Department of Optometry and Vision Sciences University of Melbourne,
Parkville, Victoria, Australia. (2)Department of Anatomy & Developmental Biology,
School of Biomedical Sciences, Monash University, Victoria, Australia. (3)School 
of Molecular Sciences, Department of Biochemistry, La Trobe Institute for
Molecular Science, La Trobe University, Bundoora, Australia. (4)Lions Eye
Institute and the Centre for Ophthalmology and Visual Sciences, University of
Western Australia, Perth, Western Australia, Australia 5Section of Immunology and
Infection, Institute of Medical Sciences, University of Aberdeen, Aberdeen,
United Ki.

Purpose: We previously have reported that application of the intracellular
toll-like receptor (TLR)-9 ligand CpG-ODN onto the injured corneal surface
induces widespread inflammation within the eye, including the retina. We tested
the hypothesis that topical application of two other intracellular TLR agonists, 
Poly I:C and R848, would cause retinal microglial activation and migration into
the subretinal space.
Methods: C57BL/6J wild-type and Cx3cr1gfp/+ mice were anesthetized and received
central corneal abrasions followed by topical application of Poly I:C (TLR3
agonist), R848 (TLR7/8 agonist), or CpG-ODN (TLR9 agonist). Eyes were imaged in
vivo by using spectral-domain optical coherence tomography to assess and quantify
vitreous cells and retinal edema. Tissues were processed for whole-mount
immunofluorescence staining or gene expression studies. Microglial activation was
determined by morphologic changes, major histocompatibility complex (MHC) class
II reactivity, and migration to the subretinal space. Expression of
proinflammatory cytokine gene IL-6, IL-1β, IFN-γ, and MCP-1 in retinal tissues
were analyzed.
Results: At 24 hours, topical treatment with CpG-ODN and R848, but not Poly I:C, 
led to altered microglial morphology. One week after CpG-ODN and R848-treatment, 
eyes exhibited vitritis and mild inner retinal edema, increased number of
subretinal Iba-1+ cells, and an increase in MHC II+ cells in the neural retina.
Proinflammatory cytokine genes were upregulated after R848 treatment, whereas in 
the CpG-ODN group, only IL-1β and MCP-1 were significantly upregulated. Retinal
microglial activation was not observed in the Poly I:C-treated group.
Conclusions: Topical application of CpG-ODN and R848, but not Poly I:C, to the
damaged corneal surface can cause activation and migration of retinal microglia.

PMID: 26574796   [PubMed - as supplied by publisher]


11. Invest Ophthalmol Vis Sci. 2015 Nov 1;56(12):7369-7376. doi:
10.1167/iovs.15-17266.

A Comparative Treatment Study of Intravitreal Voriconazole and Liposomal
Amphotericin B in an Aspergillus fumigatus Endophthalmitis Model.

Zhao J, Cheng Y, Song X, Wang C, Su G, Liu Z.

Purpose: To compare the effects of voriconazole (VCZ) and liposomal amphotericin 
B (Amp-B) in an experimental model of Aspergillus fumigatus endophthalmitis.
Methods: Thirty guinea pigs received an intravitreal injection of A. fumigatus to
induce endophthalmitis. The animals were randomly divided into three groups,
including control (0.02 mL balanced salt solution intravitreal injection) and
experimental (20 μg VCZ/0.02 mL or 20 μg liposomal Amp-B/0.02 mL intravitreal
injection) groups. Corneal opacity, aqueous flare, and vitreous opacity were
graded, and electroretinographic examinations were performed at multiple time
points. At 28 days post treatment, histopathology was performed to examine the
retinal architecture.
Results: The inflammation in the VCZ and liposomal Amp-B groups was milder than
that in the control group. Corneal opacity, aqueous flare, and vitreous opacity
scores, as well as electroretinographic recording, showed significantly less
inflammation in the VCZ group compared with the liposomal Amp-B group during the 
early and middle stages of endophthalmitis (P < 0.05). Normal histologic
structure of the retina was observed in eyes treated with VCZ and liposomal
Amp-B.
Conclusions: Both intravitreal VCZ and liposomal Amp-B were effective treatments 
for A. fumigatus-induced endophthalmitis in guinea pigs. Voriconazole was
superior to liposomal Amp-B at doses similar to the initial therapy for acute
infections. Further experimental and clinical studies are required to confirm the
efficacy of these two antifungal drugs.
Chinese Abstract.

PMID: 26574795   [PubMed - as supplied by publisher]


12. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6855-6863. doi:
10.1167/iovs.14-16291.

CYP2E1 in the Human Retinal Pigment Epithelium: Expression, Activity, and
Induction by Ethanol.

Martinez-Gil N, Flores-Bellver M, Atienzar-Aroca S, Lopez-Malo D, Urdaneta AC,
Sancho-Pelluz J, Peris-Martínez C, Bonet-Ponce L, Romero FJ, Barcia JM.

Purpose: Cytochrome p450 2E1 (CYP2E1) is a detoxifying enzyme with particular
affinity for ethanol (EtOH) expressed in several tissues. Although CYP2E1 has
been identified in human RPE, nothing is known about its metabolic activity.
Expression of CYP2E1 and activity after EtOH exposure have been studied in human 
RPE and ARPE-19 cells.
Methods: Ethanol-induced CYP2E1 mRNA expression was analyzed by RT-PCR and
quantitative PCR (qPCR) from human donor RPE as well as from ARPE-19 cells.
Expression of CYP2E1 protein was determined by Western blot. Cytoplasmic CYP2E1
location also was demonstrated by immunocytochemistry. Cell viability was studied
by the colorimetric assay XTT after EtOH treatment. Diallyl sulfide (DAS) was
used to inhibit CYP2E1 activity. The microsomal CYP2E1 activity assay was
determined by quantification of 4-nitrocatechol (4NC) formation through HPLC.
Results: Relevant CYP2E1 mRNA levels are present in human RPE. Ethanol augmented 
the formation of reactive oxygen species (ROS) in ARPE-19 cells. Expression of
CYP2E1 mRNA, CYP2E1 protein activity, and ROS production were induced by ethanol 
in a concentration-dependent manner. Interestingly, the treatment with DAS
reduced all the aforementioned increased values. The presence of CYP2E1 in both
hRPE and ARPE-19 cells reinforces the protective role of the RPE and strongly
suggests additional roles for CYP2E1 related to vision.

PMID: 26567798   [PubMed - as supplied by publisher]


13. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6847-6854. doi:
10.1167/iovs.15-17228.

Predicting Lens Diameter: Ocular Biometry With High-Resolution MRI.

Erb-Eigner K(1), Hirnschall N(2), Hackl C(2), Schmidt C(1), Asbach P(1), Findl
O(3).

Author information: 
(1)Department of Radiology, Charité-Universitätsmedizin Berlin, Campus Benjamin
Franklin, Berlin, Germany. (2)Vienna Institute for Research in Ocular Surgery
(VIROS), a Karl Landsteiner Institute, Hanusch Hospital, Vienna, Austria.
(3)Vienna Institute for Research in Ocular Surgery (VIROS), a Karl Landsteiner
Institute, Hanusch Hospital, Vienna, Austria 3Moorfields Eye Hospital NHS
Foundation Trust, London, United Kingdom.

Purpose: The aim of this study was to correlate different biometric dimensions of
the eye as measured from ocular magnetic resonance imaging (MRI) scans to predict
the lens diameter.
Methods: High-resolution ocular MRI scans of 100 eyes of 100 patients were
reviewed. Various anatomical variables of the eye such as the axial length, the
globe diameter, and the lens dimensions were measured. Also, the distances
between the ciliary sulcus and angle-to-angle were measured. A partial least
square (PLS) regression model was built to analyze which variables influence the 
model regarding the lens dimensions.
Results: Sixty-two eyes of 62 patients were included in the final analysis. The
lens diameter ratio (horizontal to vertical) was 0.93 (SD: 0.04; 0.83-1.00). The 
partial least square regression showed a significant connection (P < 0.001)
between the horizontal and vertical diameter. The partial least square regression
model that included the globe diameter and the axis length resulted in the best
prediction for the horizontal lens diameter. Similar to the horizontal lens
diameter, globe diameter was the best predictor for the vertical lens diameter
followed by the distance of the ciliary sulcus. White-to-white distance, distance
of the ciliary sulcus, and axial eye length were found to have a high influence
on the angle-to-angle distance.
Conclusions: The introduced models may serve as tools to predict the capsular bag
biometry in a preoperative setting for cataract surgery or lens refilling
procedures.

PMID: 26567797   [PubMed - as supplied by publisher]


14. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6839-6846. doi:
10.1167/iovs.15-16792.

Neuroprotective Effects of Voluntary Exercise in an Inherited Retinal
Degeneration Mouse Model.

Hanif AM(1), Lawson EC(1), Prunty M(1), Gogniat M(1), Aung MH(2), Chakraborty
R(2), Boatright JH(3), Pardue MT(3).

Author information: 
(1)Center for Visual and Neurocognitive Rehabilitation, Atlanta VA Medical
Center, Decatur, Georgia, United States. (2)Department of Ophthalmology, Emory
University School of Medicine, Atlanta, Georgia, United States. (3)Center for
Visual and Neurocognitive Rehabilitation, Atlanta VA Medical Center, Decatur,
Georgia, United States 2Department of Ophthalmology, Emory University School of
Medicine, Atlanta, Georgia, United States.

Purpose: Our previous investigations showed that involuntary treadmill exercise
is neuroprotective in a light-induced retinal degeneration mouse model, and it
may act through activation of tropomyosin-related kinase B (TrkB) receptors. This
study investigated whether voluntary running wheel exercise can be
neuroprotective in an inheritable model of the retinal degenerative disease
retinitis pigmentosa (RP), rd10 mice.
Methods: Breeding pairs of rd10 and C57BL/6J mice were given free-spinning
(active) or locked (inactive) running wheels. Pups were weaned into separate
cages with their parents' respective wheel types, and visual function was tested 
with ERG and a virtual optokinetic system at 4, 5, and 6 weeks of age. Offspring 
were killed at 6 weeks of age and retinal cross-sections were prepared for
photoreceptor nuclei counting. Additionally, separate cohorts of active and
inactive rd10 pups were injected daily for 14 days after eye opening with a
selective TrkB receptor antagonist (ANA-12) or vehicle solution and assessed as
described above.
Results: Mice in the rd10 active group exhibited significant preservation of
visual acuity, cone nuclei, and total photoreceptor nuclei number. Injection with
ANA-12 precluded the preservation of visual acuity and photoreceptor nuclei
number in rd10 mice.
Conclusions: Voluntary running partially protected against the retinal
degeneration and vision loss that otherwise occurs in the rd10 mouse model of RP.
This protection was prevented by injection of ANA-12, suggesting that TrkB
activation mediates exercise's preservation of the retina. Exercise may serve as 
an effective, clinically translational intervention against retinal degeneration.

PMCID: PMC4627249 [Available on 2016-04-01]
PMID: 26567796   [PubMed - as supplied by publisher]


15. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6823-6831. doi:
10.1167/iovs.15-17527.

Strain-Dependent Anterior Segment Dysgenesis and Progression to Glaucoma in
Col4a1 Mutant Mice.

Mao M(1), Smith RS(2), Alavi MV(1), Marchant JK(3), Cosma M(2), Libby RT(4), John
SW(5), Gould DB(1).

Author information: 
(1)Departments of Ophthalmology and Anatomy Institute for Human Genetics, UCSF
School of Medicine, San Francisco, California, United States. (2)The Jackson
Laboratory, Bar Harbor, Maine, United States. (3)The Jackson Laboratory, Bar
Harbor, Maine, United States 3Department of Anatomy and Cell Biology, Department 
of Ophthalmology, Tufts University School of Medicine, Boston, Massachusetts,
United States. (4)Flaum Eye Institute, Department of Biomedical Genetics, The
Center for Visual Sciences, University of Rochester Medical Center, Rochester,
New York, United States. (5)The Jackson Laboratory, Bar Harbor, Maine, United
States 3Department of Anatomy and Cell Biology, Department of Ophthalmology,
Tufts University School of Medicine, Boston, Massachusetts, United States 5The
Howard Hughes Medical Institute, Bar Harbor, Main.

Purpose: Mutations in the gene encoding collagen type IV alpha 1 (COL4A1) cause
multisystem disorders including anterior segment dysgenesis (ASD) and optic nerve
hypoplasia. The penetrance and severity of individual phenotypes depends on
genetic context. Here, we tested the effects of a Col4a1 mutation in two
different genetic backgrounds to compare how genetic context influences ocular
dysgenesis, IOP, and progression to glaucoma.
Methods: Col4a1 mutant mice maintained on a C57BL/6J background were crossed to
either 129S6/SvEvTac or CAST/EiJ and the F1 progeny were analyzed by slit-lamp
biomicroscopy and optical coherence tomography. We also measured IOPs and
compared tissue sections of eyes and optic nerves.
Results.: We found that the CAST/EiJ inbred strain has a relatively uniform and
profound suppression on the effects of Col4a1 mutation and that mutant CASTB6F1
mice were generally only very mildly affected. In contrast, mutant 129B6F1 mice
had more variable and severe ASD and IOP dysregulation that were associated with 
glaucomatous signs including lost or damaged retinal ganglion cell axons and
excavation of the optic nerve head.
Conclusions.: Ocular defects in Col4a1 mutant mice model ASD and glaucoma that
are observed in a subset of patients with COL4A1 mutations. We demonstrate that
different inbred strains of mice give graded severities of ASD and we detected
elevated IOP and glaucomatous damage in 129B6F1, but not CASTB6F1 mice that
carried a Col4a1 mutation. These data demonstrate that genetic context
differences are one factor that may contribute to the variable penetrance and
severity of ASD and glaucoma in patients with COL4A1 mutations.

PMCID: PMC4627250 [Available on 2016-04-01]
PMID: 26567795   [PubMed - as supplied by publisher]


16. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6810-6822. doi:
10.1167/iovs.15-16866.

Intravitreal Ciliary Neurotrophic Factor Transiently Improves Cone-Mediated
Function in a CNGB3-/- Mouse Model of Achromatopsia.

Marangoni D(1), Vijayasarathy C(2), Bush RA(2), Wei LL(3), Wen R(4), Sieving
PA(5).

Author information: 
(1)National Institute on Deafness and Other Communication Disorders, National
Institutes of Health, Bethesda, Maryland, United States 3Department of
Biotechnology and Applied Clinical Science, University of L'Aquila, L'Aquila,
Italy. (2)National Institute on Deafness and Other Communication Disorders,
National Institutes of Health, Bethesda, Maryland, United States. (3)National Eye
Institute, National Institutes of Health, Bethesda, Maryland, United States.
(4)Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami,
Miller School of Medicine, Miami, Florida, United States. (5)National Institute
on Deafness and Other Communication Disorders, National Institutes of Health,
Bethesda, Maryland, United States 2National Eye Institute, National Institutes of
Health, Bethesda, Maryland, United States.

Purpose: Ciliary neurotrophic factor (CNTF) was recently shown to augment cone
function in CNGB3 mutant achromat dogs. However, testing CNTF-releasing implant
in human CNGB3 achromats failed to show benefit. We evaluated the effects of CNTF
protein on the retinal function in an additional achromatopsia model, the
CNGB3-/- mouse.
Methods: Fifty-nine CNGB3-/- mice (postnatal day [PD] ± SD = 30 ± 7) received a
unilateral intravitreal injection of 1 or 2 μg CNTF protein, and 15 wild-type
(WT) mice (PD = 34 ± 3) received 1 μg CNTF. Retinal function was evaluated by
flash ERG and photopic flicker ERG (fERG) at 7 and 14 days after treatment.
Results: Seven days post CNTF, the photopic b-wave Vmax was significantly
increased in CNGB3-/- mice (P < 0.01), whereas it was reduced in WT mice (P <
0.05). Ciliary neurotrophic factor significantly increased the amplitude of
photopic fERG and the photopic oscillatory potentials (OPs) in CNGB3-/- mice.
Ciliary neurotrophic factor did not alter the scotopic a-wave in either CNGB3-/- 
or WT mice, but it increased the scotopic b-wave k (P < 0.01) in CNGB3-/- mice,
indicating diminished scotopic sensitivity, and reduced the scotopic b-wave Vmax 
in WT mice (P < 0.05). No difference was found in ERG parameters between 1 or 2
μg CNTF. Fourteen days after CNTF injection the ERG changes in CNGB3-/- mice were
lost.
Conclusions: Intravitreal bolus CNTF protein caused a small and transient
improvement of cone-mediated function in CNGB3-/- mice, whereas it reduced
rod-mediated function. The increase in photopic OPs and the lack of changes in
scotopic a-wave suggest a CNTF effect on the inner retina.

PMCID: PMC4627357 [Available on 2016-04-01]
PMID: 26567794   [PubMed - as supplied by publisher]


17. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6801-6809. doi:
10.1167/iovs.15-16822.

Inferring an Evolutionary Tree of Uveal Melanoma From Genomic Copy Number
Aberrations.

Singh N(1), Singh AD(2), Hide W(3).

Author information: 
(1)School of Medicine, Case Western Reserve University, Cleveland, Ohio, United
States. (2)Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States. 
(3)Biostatistics Department, Harvard School of Public Health, Boston,
Massachusetts, United States 4Sheffield Institute for Translational Neuroscience,
University of Sheffield, Sheffield, United Kingdom.

Purpose: The purpose of this study is to study the genomic evolution of primary
uveal melanoma.
Methods: Primary uveal melanoma genomic DNA was assayed on the Illumina
Human660W-Quad v1.0 DNA Analysis BeadChip. Raw signal intensity data were
quantile normalized to estimate copy number aberration with the Genome Alteration
Print algorithm. Distance between samples was calculated as the Manhattan
distance between the copy number profiles of the tumors. From the distance
matrix, a phylogenetic network (evolutionary relationship inference) was
estimated using SplitsTree4.
Results: Of the 57 tumors, one (1.8%) was discarded because of a failed assay,
and seven (12.3%) were revealed to be mixtures of several cell populations that
could not be resolved. Three clades of tumor were identified (A [59.2%], B
[32.7%], and C [6.1%]), each following a distinct evolutionary path and each
associated with metastatic status (P = 0.01). One tumor (2.0%) did not fit into
any clade. From a normal diploid melanocyte, a few tumors (clade C) lose a large 
portion of chromosome 6q, but do not develop any mutations on 8q. In an alternate
path, the vast majority of tumors (clade A and clade B [91.9%]) gain a copy of
the telomeric half of 8q. A majority of these tumors (clade A) subsequently lose 
a copy of chromosome 3, as well as gain the centromeric half of 8q. The other
tumors (clade B) gain copies of 6p, as well as regions on 11p and 22q.
Conclusions: Our data suggest that there is little overlap in the subtypes of
uveal melanoma after divergence (identified as clades A and B) and that these
distinct subtypes are not likely to crossover or transform from one major clade
to another.

PMID: 26567793   [PubMed - as supplied by publisher]


18. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6796-6800. doi:
10.1167/iovs.15-17481.

Retinal Blood Flow and Retinal Blood Oxygen Saturation in Mild to Moderate
Diabetic Retinopathy.

Tayyari F(1), Khuu LA(2), Flanagan JG(3), Singer S(2), Brent MH(2), Hudson C(3).

Author information: 
(1)Retina Research Group, School of Optometry and Vision Science, University of
Waterloo, Waterloo, Ontario, Canada. (2)Department of Ophthalmology and Vision
Sciences, University of Toronto, Toronto, Ontario, Canada. (3)Retina Research
Group, School of Optometry and Vision Science, University of Waterloo, Waterloo, 
Ontario, Canada 2Department of Ophthalmology and Vision Sciences, University of
Toronto, Toronto, Ontario, Canada.

Purpose: The aim of this study was to evaluate the relationship between retinal
blood flow (RBF) and retinal blood oxygen saturation (SO2) in mild to moderate
nonproliferative diabetic retinopathy (NPDR) and in age-matched controls.
Methods: One eye of each of 15 healthy subjects (68 ± 6 years) and 13 subjects
with mild to moderate NPDR (67 ± 10 years) was dilated. None of the patients with
NPDR had received treatment for their retinopathic changes or had any evidence of
sight-threatening characteristics. Doppler Fourier-domain optical coherence
tomography blood flow was measured using the prototype RTVue system; six separate
measurements each comprising an upper and a lower nasal pupil scan were acquired.
Six hyperspectral retinal measurements were acquired using a noninvasive
hyperspectral retinal camera (prototype H-8.5 HR Camera).
Results: Total RBF was significantly lower in NPDR when compared to controls
(42.7 ± 7.5 vs. 33.0 ± 9.2 μL/min; P = 0.004). Mean retinal arterial and venular 
SO2 were higher in NPDR than in controls (94.7 ± 2.4% vs. 92.9 ± 1.6%, P = 0.02; 
62.5 ± 5.7% vs. 56.3 ± 4.7%, P = 0.003). This study showed a correlation between 
RBF and arteriolar SO2 in both controls (r = 0.58, P = 0.02) and NPDR (r = 0.54, 
P = 0.05), but no correlation between venular RBF and venular SO2 in controls (r 
= 0.24, P = 0.83) or in NPDR (r = 0.23, P = 0.45). The arteriovenous difference
(AV difference) was lower in the NPDR group when compared to controls (30.6 ± 6
vs. 36.7 ± 5.3, P = 0.008).
Conclusions: This study found a lower total RBF and a lower AV difference in the 
NPDR group, suggesting a reduced oxygen uptake from the retina in people with
relatively early diabetic retinopathy.

PMID: 26567792   [PubMed - as supplied by publisher]


19. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6788-6795. doi:
10.1167/iovs.15-17176.

A Diagnostic Calculator for Detecting Glaucoma on the Basis of Retinal Nerve
Fiber Layer, Optic Disc, and Retinal Ganglion Cell Analysis by Optical Coherence 
Tomography.

Larrosa JM(1), Moreno-Montañés J(2), Martinez-de-la-Casa JM(3), Polo V(1),
Velázquez-Villoria Á(2), Berrozpe C(3), García-Granero M(4).

Author information: 
(1)Ophthalmology Service Hospital Universitario Miguel Servet, Faculty of
Medicine, University of Saragossa, Saragossa, Spain. (2)Department of
Ophthalmology, Clínica Universidad de Navarra, Faculty of Medicine, Universidad
of Navarra, Pamplona, Spain. (3)Ophthalmology Service, Hospital Clínico San
Carlos, Faculty of Medicine, Universidad Complutense de Madrid, Madrid, Spain.
(4)Unidad de Estadística, Department of Biochemistry and Genetics, University of 
Navarra, Pamplona, Spain.

Purpose: The purpose of this study was to develop and validate a multivariate
predictive model to detect glaucoma by using a combination of retinal nerve fiber
layer (RNFL), retinal ganglion cell-inner plexiform (GCIPL), and optic disc
parameters measured using spectral-domain optical coherence tomography (OCT).
Methods: Five hundred eyes from 500 participants and 187 eyes of another 187
participants were included in the study and validation groups, respectively.
Patients with glaucoma were classified in five groups based on visual field
damage. Sensitivity and specificity of all glaucoma OCT parameters were analyzed.
Receiver operating characteristic curves (ROC) and areas under the ROC (AUC) were
compared. Three predictive multivariate models (quantitative, qualitative, and
combined) that used a combination of the best OCT parameters were constructed. A 
diagnostic calculator was created using the combined multivariate model.
Results: The best AUC parameters were: inferior RNFL, average RNFL, vertical
cup/disc ratio, minimal GCIPL, and inferior-temporal GCIPL. Comparisons among the
parameters did not show that the GCIPL parameters were better than those of the
RNFL in early and advanced glaucoma. The highest AUC was in the combined
predictive model (0.937; 95% confidence interval, 0.911-0.957) and was
significantly (P = 0.0001) higher than the other isolated parameters considered
in early and advanced glaucoma. The validation group displayed similar results to
those of the study group.
Conclusions: Best GCIPL, RNFL, and optic disc parameters showed a similar ability
to detect glaucoma. The combined predictive formula improved the glaucoma
detection compared to the best isolated parameters evaluated. The diagnostic
calculator obtained good classification from participants in both the study and
validation groups.

PMID: 26567791   [PubMed - as supplied by publisher]


20. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6779-6787. doi:
10.1167/iovs.14-15978.

Light Exposure and Eye Growth in Childhood.

Read SA, Collins MJ, Vincent SJ.

Purpose: The purpose of this study was to examine the relationship between
objectively measured ambient light exposure and longitudinal changes in axial eye
growth in childhood.
Methods: A total of 101 children (41 myopes and 60 nonmyopes), 10 to 15 years of 
age participated in this prospective longitudinal observational study. Axial eye 
growth was determined from measurements of ocular optical biometry collected at
four study visits over an 18-month period. Each child's mean daily light exposure
was derived from two periods (each 14 days long) of objective light exposure
measurements from a wrist-worn light sensor.
Results: Over the 18-month study period, a modest but statistically significant
association between greater average daily light exposure and slower axial eye
growth was observed (P = 0.047). Other significant predictors of axial eye growth
in this population included children's refractive error group (P < 0.001), sex (P
< 0.01), and age (P < 0.001). Categorized according to their objectively measured
average daily light exposure and adjusting for potential confounders (age, sex,
baseline axial length, parental myopia, nearwork, and physical activity),
children experiencing low average daily light exposure (mean daily light
exposure: 459 ± 117 lux, annual eye growth: 0.13 mm/y) exhibited significantly
greater eye growth than children experiencing moderate (842 ± 109 lux, 0.060
mm/y), and high (1455 ± 317 lux, 0.065 mm/y) average daily light exposure levels 
(P = 0.01).
Conclusions: In this population of children, greater daily light exposure was
associated with less axial eye growth over an 18-month period. These findings
support the role of light exposure in the documented association between time
spent outdoors and childhood myopia.

PMID: 26567790   [PubMed - as supplied by publisher]


21. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6770-6778. doi:
10.1167/iovs.15-17351.

Infiltration of Plasma Cells in the Iris of Children With ANA-Positive Anterior
Uveitis.

Kalinina Ayuso V(1), van Dijk MR(2), de Boer JH(1).

Author information: 
(1)Department of Ophthalmology University Medical Center Utrecht, Utrecht, The
Netherlands. (2)Department of Pathology, University Medical Center Utrecht,
Utrecht, The Netherlands.

Purpose: We investigated inflammatory cell infiltrates in iris biopsies in
uveitis associated with juvenile idiopathic arthritis (JIA) in comparison with
other pediatric uveitis entities and noninflammatory pediatric controls.
Methods: Iridectomy specimens were obtained during elective trabeculectomy from
31 eyes of 25 patients: 12 eyes with JIA-associated uveitis, 13 eyes with other
uveitis entities, and 6 eyes with open angle nonuveitic juvenile glaucoma.
Histopathologic and immunohistochemical analyses were performed. A
semiquantitative scoring system was used with a scale ranging from 0 to 4
depending on the number of stained cells.
Results: An inflammatory infiltrate was present in 8/12 (67%) specimens with
JIA-associated uveitis. The cellular infiltrate in JIA specimens was
characterized by the presence of CD138+ plasma cells and CD68+ macrophages, while
the presence of CD20+, CD4+, and CD8+ cells was variable. Presence of plasma
cells in the inflammatory infiltrates in anterior uveitis correlated with
antinuclear autoantibody (ANA) positivity regardless of the diagnosis of JIA.
CD4+ and CD8+ T cells were not always detectable in the iris biopsies of all
childhood uveitis patients, although a slight predominance of CD4+ cells was
noted.
Conclusions: Children with ANA-positive anterior uveitis often show an infiltrate
of plasma cells, regardless of the diagnosis of JIA. The iris of JIA-associated
uveitis patients is additionally characterized by the presence of various numbers
of macrophages.

PMID: 26567789   [PubMed - as supplied by publisher]


22. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6762-6769. doi:
10.1167/iovs.15-17201.

Number of People Blind or Visually Impaired by Cataract Worldwide and in World
Regions, 1990 to 2010.

Khairallah M(1), Kahloun R(1), Bourne R(2), Limburg H(3), Flaxman SR(4), Jonas
JB(5), Keeffe J(6), Leasher J(7), Naidoo K(8), Pesudovs K(9), Price H(2), White
RA(10), Wong TY(11), Resnikoff S(12), Taylor HR(13); Vision Loss Expert Group of 
the Global Burden of Disease Study.

Author information: 
(1)Department of Ophthalmology, Fattouma Bourguiba University Hospital, Faculty
of Medicine, University of Monastir, Monastir, Tunisia. (2)Vision and Eye
Research Unit, Anglia Ruskin University, Cambridge, United Kingdom. (3)Consultant
public eye health, Health Information Services, Grootebroek, The Netherlands.
(4)School of Computer Science and Heinz College, Carnegie Mellon University,
Pittsburgh, Pennsylvania, United States. (5)Department of Ophthalmology, Medical 
Faculty Mannheim, Heidelberg University, Mannheim, Germany. (6)Department of
Ophthalmology, University of Melbourne, Melbourne, Australia. (7)Nova
Southeastern University, Fort Lauderdale, Florida, United States. (8)African
Vision Research Institute, University of Kwazulu-Natal, South Africa and Brien
Holden Vision Institute, Sydney, Australia. (9)NHMRC Centre for Clinical Eye
Research, Flinders University, Adelaide, Australia. (10)Department of Genes and
Environment, Division of Epidemiology, Norwegian Institute of Public Health,
Oslo, Norway. (11)Singapore Eye Research Institute, Singapore. (12)Brien Holden
Vision Institute, University of New South Wales, Sydney, Australia. (13)Melbourne
School of Population and Global Health, University of Melbourne, Australia.

Purpose: To estimate prevalence and number of people visually impaired or blind
due to cataract.
Methods: Based on the Global Burden of Diseases Study 2010 and ongoing literature
research, we examined how many people were affected by moderate to severe vision 
impairment (MSVI; presenting visual acuity <6/18, ≥3/60) and blindness
(presenting visual acuity <3/60) due to cataract.
Results: In 2010, of overall 32.4 million blind and 191 million vision impaired, 
10.8 million people were blind and 35.1 million were visually impaired due to
cataract. Cataract caused worldwide 33.4% of all blindness in 2010, and 18.4% of 
all MSVI. These figures were lower in the high-income regions (<15%) and higher
(>40%) in South and Southeast Asia and Oceania. From 1990 to 2010, the number of 
blind or visually impaired due to cataract decreased by 11.4% and by 20.2%,
respectively; the age-standardized global prevalence of cataract-related
blindness and MSVI reduced by 46% and 50%, respectively, and the worldwide crude 
prevalence of cataract-related blindness and MSVI reduced by 32% and 39%,
respectively. The percentage of global blindness and MSVI caused by cataract
decreased from 38.6% to 33.4%, and from 25.6% to 18.4%, respectively. This
decrease took place in almost all world regions, except East Sub-Saharan Africa.
Conclusions: In 2010, one in three blind people was blind due to cataract, and
one of six visually impaired people was visually impaired due to cataract.
Despite major improvements in terms of reduction of prevalence, cataract remains 
a major public health problem.

PMID: 26567788   [PubMed - as supplied by publisher]


23. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6754-6761. doi:
10.1167/iovs.15-17534.

NADPH Oxidase-Mediated ROS Production Determines Insulin's Action on the Retinal 
Microvasculature.

Kida T, Oku H, Horie T, Matsuo J, Kobayashi T, Fukumoto M, Ikeda T.

Purpose: To determine whether insulin induces nitric oxide (NO) formation in
retinal microvessels and to examine the effects of high glucose on the formation 
of NO.
Methods: Freshly isolated rat retinal microvessels were incubated in normal (5.5 
mM) or high (20 mM) glucose with or without insulin (100 nM). The levels of
insulin-induced NO and reactive oxygen species (ROS) in the retinal microvessels 
were determined semiquantitatively using fluorescent probes,
4,5-diaminofluorescein diacetate, and hydroethidine, respectively, and a laser
scanning confocal microscope. The insulin-induced changes of NO in rat retinal
endothelial cells and pericytes cultured at different glucose concentrations (5.5
and 25 mM) were determined using flow cytometry. Nitric oxide synthase (NOS)
protein levels were determined by Western blot analysis; intracellular levels of 
ROS were determined using fluorescence-activated cell sorting (FACS) analysis of 
ethidium fluorescence; and nicotinamide adenine dinucleotide phosphate (NADPH)
oxidase RNA expression was quantified using real-time PCR.
Results: Exposure of microvessels to insulin under normal glucose conditions led 
to a significant increase in NO levels; however, this increase was significantly 
suppressed when the microvessels were incubated under high glucose conditions.
Intracellular levels of ROS were significantly increased in both retinal
microvessels and cultured microvascular cells under high glucose conditions. The 
expression of NOS and NADPH oxidase were significantly increased in endothelial
cells and pericytes under high glucose conditions.
Conclusions: The increased formation of NO by insulin and its suppression by high
glucose conditions suggests that ROS production mediated by NADPH oxidase is
important by insulin's effect on the retinal microvasculature.

PMID: 26567787   [PubMed - as supplied by publisher]


24. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6747-6753. doi:
10.1167/iovs.15-17720.

Expression Profiling of Human Schlemm's Canal Endothelial Cells From Eyes With
and Without Glaucoma.

Cai J(1), Perkumas KM(2), Qin X(3), Hauser MA(4), Stamer WD(5), Liu Y(6).

Author information: 
(1)Department of Cellular Biology and Anatomy, Georgia Regents University,
Augusta, Georgia, United States. (2)Department of Ophthalmology, Duke University,
Durham, North Carolina, United States. (3)Duke Molecular Physiology Institute,
Duke University, Durham, North Carolina, United States 4Department of Medicine,
Duke University, Durham, North Carolina, United States. (4)Department of
Ophthalmology, Duke University, Durham, North Carolina, United States 3Duke
Molecular Physiology Institute, Duke University, Durham, North Carolina, United
States 4Department of Medicine, Duke University, Durham, North Carolina, United
State. (5)Department of Ophthalmology, Duke University, Durham, North Carolina,
United States 5Department of Biomedical Engineering, Duke University, Durham,
North Carolina, United States. (6)Department of Cellular Biology and Anatomy,
Georgia Regents University, Augusta, Georgia, United States 6James & Jean Culver 
Vision Discovery Institute, Georgia Regents University, Augusta, Georgia, United 
States 7Center for Biotechnology and Genomic Medi.

Purpose: Ocular hypertension is a major risk factor for glaucoma and the inner
wall of Schlemm's canal (SC) endothelia participates in the regulation of aqueous
humor outflow resistance. This study aimed to identify differentially expressed
genes in primary cultures of SC cells from glaucoma patients.
Methods: This study examined SC samples from three glaucoma cases and four
controls. Schlemm's canal cells were isolated from eight different postmortem
human eyes. Total RNA was extracted, labeled, and hybridized to Illumina
HumanWG-6 BeadChips containing probes for approximately 47,000 human transcripts.
After extracting the data using Illumina GenomeStudio software, the data were
normalized and analyzed using the R package limma in Bioconductor. Using Protein 
ANalysis THrough Evolutionary Relationships (PANTHER) software, gene ontology
analysis of highly expressed genes was executed in controls and glaucoma groups
separately. Pathway analysis was performed with differentially expressed genes
using WebGestalt (WEB-based GEne SeT AnaLysis Toolkit). Selected genes were
validated using droplet digital PCR (ddPCR).
Results: Gene ontology analysis indicated similar functional categories in cases 
and controls. Differential analysis identified a total of 113 genes with at least
2-fold expression changes in cases. Pathway analysis indicated significant
enrichment of genes in cell adhesion, heparin binding, glycosaminoglycan binding,
filopodium, and extracellular matrix remodeling. Eighteen selected genes with
differential expression were successfully validated using ddPCR.
Conclusions: This study represents the first genome-wide expression study of
human primary SC cells from glaucoma patients and provides a potential list of
targets regulating SC cell stiffness and pore formation, eventually the outflow
resistance in glaucoma individuals.

PMCID: PMC4614909 [Available on 2016-04-01]
PMID: 26567786   [PubMed - as supplied by publisher]


25. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6740-6746. doi:
10.1167/iovs.15-17656.

Two-Photon Fluorescence Microscopy for Determination of the Riboflavin
Concentration in the Anterior Corneal Stroma When Using the Dresden Protocol.

Seiler TG(1), Ehmke T(2), Fischinger I(1), Zapp D(3), Stachs O(4), Seiler T(5),
Heisterkamp A(6).

Author information: 
(1)Klinik und Poliklinik für Augenheilkunde, Technische Universität München,
Munich, Germany 2Institut für Refraktive und Ophthalmo-Chirurgie (IROC), Zurich, 
Switzerland. (2)Laser Zentrum Hannover e.V., Hanover, Germany. (3)Klinik und
Poliklinik für Augenheilkunde, Technische Universität München, Munich, Germany.
(4)Department of Ophthalmology, Rostock University Medical Center, Rostock,
Germany. (5)Institut für Refraktive und Ophthalmo-Chirurgie (IROC), Zurich,
Switzerland. (6)Laser Zentrum Hannover e.V., Hanover, Germany 5Institut für
Quantenoptik, Leibniz Universität Hannover, Hanover, Germany.

Purpose: To determine the riboflavin concentration gradient in the anterior
corneal stroma when using the Dresden protocol with different dextran solutions.
Methods: Three different groups of porcine corneas, five each, were compared
regarding the riboflavin concentration in the anterior stroma. Before all
experiments, stable hydration conditions were established for the corresponding
solution. All groups were treated with 0.1% riboflavin in different dextran
solutions (15%, 16%, 20%). After imbibition, two-photon microscopy was used to
determine fluorescence intensity. For signal attenuation and concentration
determination corneas were saturated and measured a second time by two-photon
microscopy. Additionally, the distribution was calculated mathematically and
compared to the empiric results.
Results: Riboflavin concentration is decreasing with depth for all dextran
solutions. A nearly constant concentration could be determined over the first 75 
μm. Analysis of the fit functions leads to diffusion coefficients of D = 2.97 ×
10-7 cm2/s for the 15% dextran solution, D = 2.34 × 10-7 cm2/s for the 16%
dextran solution, and D = 1.28 × 10-7 cm2/s for the 20% dextran solution. The
riboflavin gradients of the 20% dextran group were statistically significantly
different from 15% dextran starting at a depth of 220 μm and deeper (P = 0.047). 
The 16% dextran group differed statistically at a depth of 250 μm and deeper (P =
0.047). These results show a significant difference to those published
previously.
Conclusions: With correct settings two-photon microscopy is a precise way to
determine the concentration of riboflavin in cornea. The measured gradient is
excellently fit by a Gaussian distribution, which comes out as a solution of
Fick's second law.

PMID: 26567785   [PubMed - as supplied by publisher]


26. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6734-6739. doi:
10.1167/iovs.15-17443.

Visualization of Nerve Fiber Orientations in the Human Optic Chiasm Using
Photomicrographic Image Analysis.

Jain NS(1), Jain SV(1), Wang X(2), Neely AJ(2), Tahtali M(2), Jain S(3), Lueck
CJ(4).

Author information: 
(1)Department of Neurology, The Canberra Hospital, Canberra, Australian Capital
Territory, Australia 2Faculty of Medicine, University of New South Wales,
Kensington, Sydney, New South Wales, Australia. (2)School of Engineering and
Information Technology, University of New South Wales, Canberra, Australian
Capital Territory, Australia. (3)Department of Anatomical Pathology, The Canberra
Hospital, Canberra, Australian Capital Territory, Australia 5Australian National 
University Medical School, Canberra, Australian Capital Territory, Australia.
(4)Department of Neurology, The Canberra Hospital, Canberra, Australian Capital
Territory, Australia 5Australian National University Medical School, Canberra,
Australian Capital Territory, Australia.

Purpose: Hemidecussation of fibers entering the optic chiasm from the optic
nerves is well recognized. The reason why bitemporal hemianopia results from
chiasmal compression has not been fully explained. There is still a paucity of
data relating to the precise details of the routes that the nerve fibers take
through the chiasm and, in particular, where and how nerve fibers cross each
other. This information is important to understanding why crossing fibers are
selectively damaged as a result of chiasmal compression.
Methods: An optic chiasm obtained at postmortem was fixed, stained, and sectioned
to allow high-resolution photomicrographs to be taken. The photomicrographs were 
integrated to allow regions of interest across entire sections to be analyzed for
fiber direction and crossing.
Results: The results confirmed that fibers from the temporal retina pass directly
backward in the lateral chiasm to the optic tract, whereas fibers from the nasal 
retina cross to the contralateral optic tract. Crossings take place in the
paracentral regions of the chiasm rather than in the center of the chiasm (where 
the nerve fibers are traveling mostly in parallel). The paracentral crossing
regions are distributed in a largely postero-superior to antero-inferior
arrangement.
Conclusions: These findings clarify the precise locations and crossing angles of 
crossing nerve fibers in the chiasm. This information may help explain the
clinical observation of junctional scotoma and will provide a much better basis
for structural modeling of chiasmal compression which, in turn, will improve our 
understanding of how and why bitemporal hemianopia occurs.

PMID: 26567784   [PubMed - as supplied by publisher]


27. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6724-6733. doi:
10.1167/iovs.15-17042.

Differentiation of Diabetic Macular Edema From Pseudophakic Cystoid Macular Edema
by Spectral-Domain Optical Coherence Tomography.

Munk MR(1), Jampol LM(2), Simader C(3), Huf W(4), Mittermüller TJ(5), Jaffe
GJ(6), Schmidt-Erfurth U(5).

Author information: 
(1)Department of Ophthalmology Medical University of Vienna, Vienna, Austria
2Department of Ophthalmology, Northwestern University, Feinberg School of
Medicine, Chicago, Illinois, United States 3Department of Ophthalmology,
Inselspital, University Hospital B. (2)Department of Ophthalmology, Northwestern 
University, Feinberg School of Medicine, Chicago, Illinois, United States.
(3)Department of Ophthalmology Medical University of Vienna, Vienna, Austria
4Reading Center Vienna, Medical University of Vienna, Vienna, Austria. (4)Center 
for Medical Physics and Biomedical Engineering, Medical University of Vienna,
Vienna, Austria. (5)Department of Ophthalmology Medical University of Vienna,
Vienna, Austria. (6)Duke Eye Center, Duke Reading Center, Duke University School 
of Medicine, Durham, North Carolina, United States.

Purpose: To differentiate diabetic macular edema (DME) from pseudophakic cystoid 
macular edema (PCME) based solely on spectral-domain optical coherence tomography
(SD-OCT).
Methods: This cross-sectional study included 134 participants: 49 with PCME, 60
with DME, and 25 with diabetic retinopathy (DR) and ME after cataract surgery.
First, two unmasked experts classified the 25 DR patients after cataract surgery 
as either DME, PCME, or mixed-pattern based on SD-OCT and color-fundus
photography. Then all 134 patients were divided into two datasets and graded by
two masked readers according to a standardized reading-protocol. Accuracy of the 
masked readers to differentiate the diseases based on SD-OCT parameters was
tested. Parallel to the masked readers, a computer-based algorithm was
established using support vector machine (SVM) classifiers to automatically
differentiate disease entities.
Results: The masked readers assigned 92.5% SD-OCT images to the correct clinical 
diagnose. The classifier-accuracy trained and tested on dataset 1 was 95.8%. The 
classifier-accuracy trained on dataset 1 and tested on dataset 2 to differentiate
PCME from DME was 90.2%. The classifier-accuracy trained and tested on dataset 2 
to differentiate all three diseases was 85.5%. In particular, higher
central-retinal thickness/retinal-volume ratio, absence of an
epiretinal-membrane, and solely inner nuclear layer (INL)-cysts indicated PCME,
whereas higher outer nuclear layer (ONL)/INL ratio, the absence of subretinal
fluid, presence of hard exudates, microaneurysms, and ganglion cell layer and/or 
retinal nerve fiber layer cysts strongly favored DME in this model.
Conclusions: Based on the evaluation of SD-OCT, PCME can be differentiated from
DME by masked reader evaluation, and by automated analysis, even in DR patients
with ME after cataract surgery. The automated classifier may help to
independently differentiate these two disease entities and is made publicly
available.

PMID: 26567783   [PubMed - as supplied by publisher]


28. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6714-6723. doi:
10.1167/iovs.15-17364.

Enriched Cultures of Retinal Cells From BJNhem20 Human Embryonic Stem Cell Line
of Indian Origin.

Mariappan I(1), Maddileti S(1), Joseph P(1), Siamwala JH(2), Vauhini V(1).

Author information: 
(1)Sudhakar and Sreekanth Ravi Stem Cell Biology Laboratory Brien Holden Eye
Research Centre, Champalimaud Translational Centre for Eye Research, Hyderabad
Eye Research Foundation, L V Prasad Eye Institute, Hyderabad, India. (2)Sudhakar 
and Sreekanth Ravi Stem Cell Biology Laboratory Brien Holden Eye Research Centre,
Champalimaud Translational Centre for Eye Research, Hyderabad Eye Research
Foundation, L V Prasad Eye Institute, Hyderabad, India 2University of California,
San Die.

Purpose: To test the retinal differentiation potential and to establish an
optimized protocol for enriching retinal cells from an Indian origin, human
embryonic stem cell (hESC) line, BJNhem20.
Methods: The BJNhem20 cells were cultured and expanded under feeder-free culture 
conditions. Differentiation was initiated by embryoid body (EB) formation and
were cultured on Matrigel in neural induction medium (NIM) for 1 week and further
maintained in retinal differentiation medium (RDM). After 1 month, the
neuro-retinal progenitor clusters located at the center of pigmented retinal
patches were picked and cultured as suspended neurospheres in RDM for 3 days and 
subsequently on Matrigel in neuro-retinal medium. The mildly pigmented, immature 
retinal pigmented epithelial (RPE) cells were picked separately and cultured on
Matrigel in RPE medium (RPEM). After 1 week, the confluent neuro-retinal and RPE 
cultures were maintained in RDM for 2 to 3 months and characterized by
immunofluorescence and RT-PCR.
Results: The BJNhem20 cells efficiently differentiated into both neuro-retinal
and RPE cells. The early retinal progenitors expressed Nestin, GFAP, Pax6, Rx,
MitfA, Chx10, and Otx2. Neuro-retinal cells expressed the neural markers, Map2,
β-III tubulin, acetylated tubulin and photoreceptor-specific markers, Crx,
rhodopsin, recoverin, calbindin, PKC, NeuroD1, RLBP1, rhodopsin kinase, PDE6A,
and PDE6C. Mature RPE cells developed intense pigmentation within 3 months and
showed ZO-1 and Phalloidin staining at cell-cell junctions and expressed RPE65,
tyrosinase, bestrophin1, Mertk, and displayed phagocytic activity.
Conclusions: This study confirms the retinal differentiation potential of
BJNhem20 cells and describes an optimized protocol to generate enriched
populations of neuro-retinal and RPE cells.

PMID: 26567782   [PubMed - as supplied by publisher]


29. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6654-6662. doi:
10.1167/iovs.15-17151.

Time-Resolved Ultra-High Resolution Optical Coherence Tomography for Real-Time
Monitoring of Selective Retina Therapy.

Steiner P(1), Ebneter A(2), Berger LE(2), Zinkernagel M(2), Považay B(3), Meier
C(3), Kowal JH(4), Framme C(5), Brinkmann R(6), Wolf S(2), Sznitman R(7).

Author information: 
(1)ARTORG Center, University of Bern, Bern, Switzerland 2HuCE OptoLab, Berne
University of Applied Sciences, Biel, Switzerland. (2)Department of
Ophthalmology, Inselspital, Bern, Switzerland. (3)HuCE OptoLab, Berne University 
of Applied Sciences, Biel, Switzerland. (4)ARTORG Center, University of Bern,
Bern, Switzerland. (5)Department for Ophthalmology, Medizinische Hochschule
Hannover, Hannover, Germany. (6)Institute of Biomedical Optics, University of
Lübeck, Lübeck, Germany 6Medical Laser Center Lübeck GmbH, Lübeck, Germany.
(7)ARTORG Center, University of Bern, Bern, Switzerland 3Department of
Ophthalmology, Inselspital, Bern, Switzerland.

Purpose: Selective retina therapy (SRT) is a novel treatment for retinal
pathologies, solely targeting the RPE. During SRT, the detection of an immediate 
tissue reaction is challenging, as tissue effects remain limited to intracellular
RPE photodisruption. Time-resolved ultra-high axial resolution optical coherence 
tomography (OCT) is thus evaluated for the monitoring of dynamic optical changes 
at and around the RPE during SRT.
Methods: An experimental OCT system with an ultra-high axial resolution of 1.78
μm was combined with an SRT system and time-resolved OCT M-scans of the target
area were recorded from four patients undergoing SRT. Optical coherence
tomography scans were analyzed and OCT morphology was correlated with findings in
fluorescein angiography, fundus photography, and cross-sectional OCT.
Results: In cases in which the irradiation caused RPE damage proven by
fluorescein angiography, the lesions were well discernible in time-resolved OCT
images but remained invisible in fundus photography and cross-sectional OCT
acquired after treatment. If RPE damage was introduced, all applied SRT pulses
led to detectable signal changes in the time-resolved OCT images. The extent of
optical signal variation seen in the OCT data appeared to scale with the applied 
SRT pulse energy.
Conclusions: The first clinical results proved that successful SRT irradiation
induces detectable changes in the OCT M-scan signal while it remains invisible in
conventional ophthalmoscopic imaging. Thus, real-time high-resolution OCT is a
promising modality to monitor and analyze tissue effects introduced by selective 
retina therapy and may be used to guide SRT in an automatic feedback mode
(www.swissmedic.ch number, 2011-MD-0006).

PMID: 26567781   [PubMed - as supplied by publisher]


30. Invest Ophthalmol Vis Sci. 2015 Oct 1;56(11):6573-6580. doi:
10.1167/iovs.15-17635.

The Ability of SD-OCT to Differentiate Early Glaucoma With High Myopia From
Highly Myopic Controls and Nonhighly Myopic Controls.

Akashi A, Kanamori A, Ueda K, Inoue Y, Yamada Y, Nakamura M.

Purpose: Optical coherence tomography (OCT) instruments do not embed a normative 
database from highly myopic normal (HMN) eyes. The abilities of three OCT
instruments to detect early glaucoma with high myopia were compared using the two
controls with or without high myopia.
Methods: A total of 52 early glaucomatous eyes (mean deviation > -6.0 dB) with
high myopia (spherical equivalent ≤ -6.0 diopters [HMG]), 54 HMN eyes, and 90
nonhighly myopic normal (NHMN) eyes were enrolled. Each participant was imaged
using Cirrus, RTVue, and Topcon 3D OCT to evaluate the thicknesses of the
circumpapillary retinal nerve fiber layer (cpRNFL), the macular retinal nerve
fiber layer (mRNFL), ganglion cell layer + inner plexiform layer (GCL/IPL), and
mRNFL + GCL/IPL (GCC). The covariate-adjusted areas under the receiver operating 
characteristic curves (AUCs) for detecting HMG were compared among the
instruments and between the two normal groups (HMN or NHMN).
Results: Highly myopic normal eyes showed higher AUCs for the temporal quadrant
cpRNFL thickness but lower AUCs for the superior and inferior RNFL thicknesses
compared with NHMN. We found the AUCs for the GCC thickness showed no significant
difference between the two control groups, but the GCL/IPL and mRNFL thicknesses 
had differences.
Conclusions: The abilities of the three OCT instruments to detect early
glaucomatous eyes with high myopia were different if the normal eyes were
associated with high myopia or not. A normative database that includes data from 
patients with high myopia should be established for accurate diagnosis of
glaucoma with high myopia. (www.umin.ac.jp/ctr number, UMIN000006900.).

PMID: 26567476   [PubMed - as supplied by publisher]