Choriocapillary Flow Correlation with Axial Length in High Myopia - An Angiography Study with Optical Coherence Tomography.

PURPOSE: To investigate parameters of retinal and choroidal microcirculation quantitatively with optical coherence tomography angiography (OCTA) in high myopic children, and to explore potential correlations with age, axial length (AL), spherical equivalent (SE), and central retinal thickness (CRT). METHODS: En face angiograms were generated with an OCTA device and evaluated with automated density and flow analyzer algorithms. Perfusion parameters were correlated with age, AL, SE, and CRT using Spearman's rank correlation analysis. Repeatability and reproducibility of perfusion parameter measurements were calculated in a high myopic cohort. RESULTS: Repeatability and reproducibility of OCTA measurements were good, ranging from 3.6 - 6.5%. Strong positive correlation was identified between age and CRT (rho = 0.673, p = 0.00) as well as between AL and SE (rho = 0.844, p = 0.00). There was a strong negative correlation between AL and choriocapillary flow density (CCFD) (rho = - 0.612, p = 0.00), and a moderate negative correlation between age and superficial parafoveal retinal vessel density (SPRVD) as well as CCFD (rho = - 0.497, p = 0.013 and rho = - 0.483, p = 0.023, respectively). CONCLUSION: OCTA appears to be a reliable tool for the quantitative investigation of retinal and choroidal microcirculation in a high myopic pediatric cohort. CCFD reduction was associated with increasing AL in this cohort.

Myopia-26, the female-limited form of early-onset high myopia, occurring in a European family.

BACKGROUND: Female-limited early-onset high myopia, also called Myopia-26 is a rare monogenic disorder characterized by severe short sightedness starting in early childhood and progressing to blindness potentially by the middle ages. Despite the X-linked locus of the mutated ARR3 gene, the disease paradoxically affects females only, with males being asymptomatic carriers. Previously, this disease has only been observed in Asian families and has not gone through detailed investigation concerning collateral symptoms or pathogenesis. RESULTS: We found a large Hungarian family displaying female-limited early-onset high myopia. Whole exome sequencing of two individuals identified a novel nonsense mutation (c.214C>T, p.Arg72*) in the ARR3 gene. We carried out basic ophthalmological testing for 18 family members, as well as detailed ophthalmological examination (intraocular pressure, axial length, fundus appearance, optical coherence tomography, visual field- testing) as well as colour vision- and electrophysiology tests (standard and multifocal electroretinography, pattern electroretinography and visual evoked potentials) for eight individuals. Ophthalmological examinations did not reveal any signs of cone dystrophy as opposed to animal models. Electrophysiology and colour vision tests similarly did not evidence a general cone system alteration, rather a central macular dysfunction affecting both the inner and outer (postreceptoral and receptoral) retinal structures in all patients with ARR3 mutation. CONCLUSIONS: This is the first description of a Caucasian family displaying Myopia-26. We present two hypotheses that could potentially explain the pathomechanism of this disease.

Cellular Factor XIII, a Transglutaminase in Human Corneal Keratocytes.

Cellular factor XIII (cFXIII, FXIII-A2), a transglutaminase, has been demonstrated in a few cell types. Its main function is to cross-link proteins by isopeptide bonds. Here, we investigated the presence of cFXIII in cells of human cornea. Tissue sections of the cornea were immunostained for FXIII-A in combination with staining for CD34 antigen or isopeptide cross-links. Isolated corneal keratocytes were also evaluated by immunofluorescent microscopy and flow cytometry. FXIII-A in the corneal stroma was quantified by Western blotting. FXIII-A mRNA was detected by RT-qPCR. The cornea of FXIII-A-deficient patients was evaluated by cornea topography. FXIII-A was detected in 68 ± 13% of CD34+ keratocytes. Their distribution in the corneal stroma was unequal; they were most abundant in the subepithelial tertile. cFXIII was of cytoplasmic localization. In the stroma, 3.64 ng cFXIII/mg protein was measured. The synthesis of cFXIII by keratocytes was confirmed by RT-qPCR. Isopeptide cross-links were detected above, but not within the corneal stroma. Slight abnormality of the cornea was detected in six out of nine FXIII-A-deficient patients. The presence of cFXIII in human keratocytes was established for the first time. cFXIII might be involved in maintaining the stability of the cornea and in the corneal wound healing process.

Interaction of factor XIII subunits.

Coagulation factor XIII (FXIII) is a heterotetramer consisting of 2 catalytic A subunits (FXIII-A2) and 2 protective/inhibitory B subunits (FXIII-B2). FXIII-B, a mosaic protein consisting of 10 sushi domains, significantly prolongs the lifespan of catalytic subunits in the circulation and prevents their slow progressive activation in plasmatic conditions. In this study, the biochemistry of the interaction between the 2 FXIII subunits was investigated. Using a surface plasmon resonance technique and an enzyme-linked immunosorbent assay-type binding assay, the equilibrium dissociation constant (Kd) for the interaction was established in the range of 10(-10) M. Based on the measured Kd, it was calculated that in plasma approximately 1% of FXIII-A2 should be in free form. This value was confirmed experimentally by measuring FXIII-A2 in plasma samples immunodepleted of FXIII-A2B2. Free plasma FXIII-A2 is functionally active, and when activated by thrombin and Ca(2+), it can cross-link fibrin. In cerebrospinal fluid and tears with much lower FXIII subunit concentrations, >80% of FXIII-A2 existed in free form. A monoclonal anti-FXIII-B antibody that prevented the interaction between the 2 subunits reacted with the recombinant combined first and second sushi domains of FXIII-B, and its epitope was localized to the peptide spanning positions 96 to 103 in the second sushi domain.

Factor XIII subunits in human tears; their highly elevated levels following penetrating keratoplasty.

BACKGROUND: As blood coagulation factor XIII (FXIII) is of high importance in wound healing, we determined the concentrations of FXIII A and B subunits (FXIII-A and FXIII-B) and their complex (FXIII-A(2)B(2)) in normal tears and in tears from patients undergoing penetrating keratoplasty (PKP). METHODS: FXIII complex and subunit concentrations were measured by highly sensitive chemiluminescent ELISAs in tears from 60 healthy volunteers and from 31 patients undergoing corneal transplantation. RESULTS: In non-stimulated tears from healthy volunteers, low but consistent amounts of FXIII-A and FXIII-B (medians: 2.13 µg/L and 7.22 µg/L, respectively) were measured, mostly in non-complexed form. Following stimulation of tear secretion FXIII levels moderately decreased, but if normalized to protein concentration they did not change. One day after PKP FXIII levels became highly elevated, then gradually decreased, but even on day 7 significantly exceeded pre-surgery values. The elevation of tear FXIII levels was significantly higher in PKP patients who later developed neovascularization of donor cornea. CONCLUSIONS: FXIII subunits are low concentration components of normal tear. The striking elevation of FXIII subunit and FXIII-A(2)B(2) concentrations after PKP suggests the involvement of FXIII in corneal wound healing. Perioperatively measured high FXIII levels in tears seem to represent a risk of neovascularization.

A highly sensitive chemiluminescence immunoassay for the measurement of coagulation factor XIII subunits and their complex in tears.

Blood coagulation factor XIII (FXIII) is a tetramer (A(2)B(2)) consisting of catalytic FXIII-A and carrier/protective FXIII-B subunits. Besides stabilizing fibrin and protecting it from fibrinolysis, FXIII is also involved in wound healing and angiogenesis. The latter functions could be important in body fluids other than blood. In this study highly sensitive chemiluminescent ELISAs were developed to measure low concentrations of FXIII-A, FXIII-B and FXIII-A(2)B(2) antigen levels in tiny volumes of tear. For all three assays the limit of quantitation was below 0.02 microg/L, within laboratory imprecision did not exceed 12% and the recovery was close to 100%. The calibration curves showed an excellent fitting using second order polynomial equation. In non-stimulated tears from 40 healthy volunteers a low, but consistent amount of FXIII-A and FXIII-B (medians: 1.74 microg/L and 5.73 microg/L, respectively) was measured, mostly in free non-complexed form. The respective median values normalized for tear protein concentration were 0.23 microg FXIII-A/g and 0.85 microg FXIII-B/g. There was no gender and age difference and the values showed non-Gaussian distribution. The results prove that FXIII-A and FXIII-B are low concentration components of tear proteome, which in normal conditions might play a role in the repair of corneal micro-injuries. The developed methods provide a tool for monitoring FXIII subunit and complex levels in pathological conditions.