Evaluation of Bispecific T-Cell Engagers Targeting Murine Cytomegalovirus.

Human cytomegalovirus is a ubiquitous herpesvirus that, while latent in most individuals, poses a great risk to immunocompromised patients. In contrast to directly acting traditional antiviral drugs, such as ganciclovir, we aim to emulate a physiological infection control using T cells. For this, we constructed several bispecific T-cell engager (BiTE) constructs targeting different viral glycoproteins of the murine cytomegalovirus and evaluated them in vitro for their efficacy. To isolate the target specific effect without viral immune evasion, we established stable reporter cell lines expressing the viral target glycoprotein B, and the glycoprotein complexes gN-gM and gH-gL, as well as nano-luciferase (nLuc). First, we evaluated binding capacities using flow cytometry and established killing assays, measuring nLuc-release upon cell lysis. All BiTE constructs proved to be functional mediators for T-cell recruitment and will allow a proof of concept for this treatment option. This might pave the way for strikingly safer immunosuppression in vulnerable patient groups.

Dynamics of structural reversal in Bruch's membrane opening-based morphometrics after glaucoma drainage device surgery.

PURPOSE: Structural reversal of disc cupping is a known phenomenon after trabeculectomy. The aim of this retrospective, longitudinal, cross-sectional analysis was to evaluate the postoperative dynamics of Bruch's membrane opening-based morphometrics of the optic nerve head following glaucoma drainage device surgery. METHODS: Forty-three eyes, treated by glaucoma drainage device surgery, were included in the study. Individual changes in the spectral domain optic coherence tomography (SD-OCT) parameters Bruch's membrane opening minimum rim width (BMO-MRW), Bruch's membrane opening minimum rim area (BMO-MRA) and peripapillary retinal nerve fiber layer (RNFL) thickness as well as mean defect in 30-2 perimetry were analyzed. Changes were correlated to postoperative intraocular pressure levels over time. Available follow-up visits were aggregated and grouped into a short-term follow-up (20 to 180 days after surgery), a midterm follow-up (181 to 360 days after surgery) and a long-term follow-up (more than 360 days after surgery). RESULTS: In short-term follow-up, BMO-MRW and BMO-MRA increased significantly (p <= 0.034). This increase correlated negatively with the intraocular pressure at the time of the follow-up (Pearson's rho = - 0.49; p = 0.039). From 6 months after surgery on, there was no statistically significant change in BMO-MRW and BMO-MRA (p >= 0.207). RNFL thickness and mean defect of 30-2 perimetry showed no significant changes after GDD implantation (p >= 0.189). CONCLUSIONS: Lowering of intraocular pressure by glaucoma drainage device surgery leads to an increase of Bruch's membrane opening based parameters in the first 6 months after surgery. These changes have to be taken into account when evaluating patients' longitudinal follow-up after glaucoma drainage device implantation.

[Excision and tectonic covering of a vascular leiomyoma of the ciliary body with large anterior staphyloma using scleral patch graft]. / Exzision und tektonische Deckung eines vaskulären Leiomyoms des Ziliarkörpers mit großem anteriorem Staphylom mittels Sklerapatch.

Benign tumors of the ciliary body are rare and may potentially be confused with uveal melanoma in clinical routine. Clinical findings rarely allow for safe differentiation. Hence primary excisions are frequent. We report the case of a patient with a benign ciliary tumor, which was observed over a total of 13 years and eventually underwent a local tumor resection.

Evaluation of two-dimensional Bruch's membrane opening minimum rim area for glaucoma diagnostics in a large patient cohort.

PURPOSE: To characterize the two-dimensional parameter Bruch's membrane opening minimum rim area (BMO-MRA) in spectral domain optical coherence tomography (SD-OCT) of the optic nerve head (ONH) compared to minimum rim width (BMO-MRW) and retinal nerve fibre layer (RNFL) thickness in a large patient cohort. METHODS: Case-control, cross-sectional study of 705 eyes of 445 participants. A total of 449 eyes with glaucoma, 67 eyes with ocular hypertension and 189 healthy controls, underwent SD-OCT and confocal laser scanning tomography (CSLT), visual field testing and clinical examination. Morphometric ONH parameters, visual field function and diagnostic power were compared. Main outcome measures were SD-OCT-derived BMO-MRA, BMO-MRW, RNFL thickness and CSLT-derived rim area (DM-RA). RESULTS: Mean ONH area was 2.11 ± 0.57 mm2 ; mean BMO area was 1.89 ± 0.45 mm2 . Correlation of mean deviation in visual field to morphometric parameters was ρ = 0.70, (p < 0.001) for RNFL thickness, ρ = 0.68 (p < 0.001) for BMO-MRA, ρ = 0.66 (p < 0.001) for BMO-MRW. These correlations were not significantly different (p > 0.05), while DM-RA correlated significantly worse (ρ = 0.55; p < 0.001). In receiver operating characteristics, the calculated area under the curve (AUC) and sensitivity at 90% specificity to differentiate glaucoma were 0.87% and 70.1% for BMO-MRA, 0.86% and 68.1% for RNFL thickness, 0.84% and 66.0% for BMO-MRW, 0.82% and 51.3% for DM-RA. CONCLUSIONS: In a heterogenous clinical cohort of glaucoma patients, all analysed SD-OCT parameters excel DM-RA of CSLT. The two-dimensional parameter BMO-MRA shows comparable levels of diagnostic power to detect glaucoma compared to established parameters BMO-MRW and RNFL thickness. Given higher comparability between ONH sizes, BMO-MRA might become an additional standard tool in SD-OCT imaging for glaucoma.

Onset of Retinal Pigment Epithelium Atrophy Subsequent to Anti-VEGF Therapy in Patients with Neovascular Age-Related Macular Degeneration.

PURPOSE: The aim of this study was to evaluate risk factors for the development of retinal pigment epithelium (RPE) atrophy in patients with neovascular age-related macular degeneration (nAMD). PROCEDURES: This post hoc analysis of the prospective RESPONSE study includes 52 therapy-naive nAMD patients without baseline RPE atrophy, who were treated with ≥9 anti-vascular endothelial growth factor (VEGF) injections for ≥3 years. RPE atrophy was assessed via multimodal imaging. Baseline aqueous VEGF and serum complement levels (C3d/C3) were measured. Risk factors for atrophy development were evaluated via logistic regression analysis. RESULTS: Atrophy onset was significantly associated with the duration of nAMD (mean 5.34 years; odds ratio = 1.83, p = 0.012). Anti-VEGF injection number, age, C3d/C3 ratio, baseline intraocular VEGF, or delay to the first treatment had no influence on RPE atrophy. CONCLUSIONS: The duration of treatment-requiring nAMD was identified as primary risk factor for the onset of concomitant RPE atrophy after commencing therapy. Targeting concomitant atrophy in nAMD patients might improve the long-term prognosis of the disease.

Structural Reversal of Disc Cupping After Trabeculectomy Alters Bruch Membrane Opening-Based Parameters to Assess Neuroretinal Rim.

OBJECTIVE: To assess the impact of trabeculectomy for glaucoma on morphometric neuroretinal parameters of the optic nerve head (ONH) using spectral-domain optical coherence tomography (SD-OCT). DESIGN: Retrospective, interventional case series. METHODS: Participants: Eighty-eight eyes of 88 patients who underwent trabeculectomy with mitomycin C in 2016. INTERVENTION: All patients underwent trabeculectomy in 1 eye (study eye) and had evaluable SD-OCT examinations of the ONH to measure neuroretinal tissue before and at least at 1 of the 3-, 6-, and 12-month follow-up time points after surgery. MAIN OUTCOME MEASURES: Longitudinal change in Bruch membrane opening minimum rim width (BMO-MRW), Bruch membrane opening minimum rim area (BMO-MRA), peripapillary retinal nerve fiber layer (RNFL) thickness, intraocular pressure (IOP), and mean deviation in perimetry. RESULTS: In study eyes, BMO-MRW significantly increased postsurgically comparing baseline and follow-up examinations at 3 months (P = .012), at 6 months (P = .007), and at 1 year (P = .010) after trabeculectomy. The increase in BMO-MRW 6 months after surgery correlated with IOP reduction (r = 0.48; P = .001). BMO-MRA showed an equal increase (P ≤ .034). RNFL thickness remained stable between baseline and follow-up at 3, 6, and 12 months and showed a moderate loss after 18 months (P = .021) of follow-up. CONCLUSIONS: Structural reversal of disc cupping after trabeculectomy markedly influences Bruch membrane opening-based parameters for up to more than 1 year. Improvement in morphometry seems to correlate with the reduction of IOP while visual field function appears not to be influenced. In longitudinal follow-up of glaucoma patients by SD-OCT, evaluation of BMO-based parameters necessitates to reflect bias caused by surgery.

The Optimal Diameter for Circumpapillary Retinal Nerve Fiber Layer Thickness Measurement by SD-OCT in Glaucoma.

PURPOSE: To evaluate the diagnostic power for glaucoma detection using circumpapillary retinal nerve fiber layer (cRNFL) thickness in 3 diameter sizes from the center of the optic nerve head (ONH) by spectral domain optical coherence tomography. PATIENTS AND METHODS: In this cross-sectional study, case-control, 437 eyes diagnosed with glaucoma and 183 eyes of healthy controls underwent spectral domain optical coherence tomography of the ONH, visual field testing, and a clinical examination. cRNFL was measured by circular scans in 3.5 (C1), 4.1 (C2), and 4.7 mm (C3) distance from the center of the ONH. Receiver-operator characteristic analysis was used to assess diagnostic power to detect glaucoma; furthermore, patient-specific maximum localized damage was analyzed. RESULTS: In C1, mean global cRNFL was 70.03±18.2 µm in glaucomatous eyes and 93.46±9.9 µm in controls. Respectively, cRNFL in C2 was 61.39±14.9 and 80.43±8.4 µm as well as 55.25±12.8 and 70.70±6.7 µm in C3. Using receiver-operator characteristic analysis, the area under the curve (AUC) for cRNFL was 0.855 in C1, 0.850 in C2, and 0.843 in C3. Mean AUCs in ONH sectors ranged from 0.699 to 0.846 and did not exceed AUC of the best scoring global parameter. CONCLUSIONS: Comparing 3.5, 4.1, and 4.7 mm diameters for cRNFL measurement, the inner circle at 3.5 mm distance showed the highest AUC to differentiate glaucoma from healthy controls. However, levels of diagnostic power from wider circular scans were not significantly different and were comparable. Moreover, sectorial cRNFL measurements were nonsuperior. The use of the ONH sector with the highest localized damage seems not to increase diagnostic power.

Intraoperative changes in corneal structure during excimer laser phototherapeutic keratectomy (PTK) assessed by intraoperative optical coherence tomography.

PURPOSE: Excimer laser phototherapeutic keratectomy (PTK) is a safe treatment for superficial corneal opacities, e.g., in corneal dystrophies or degenerations. Nevertheless, no standardized treatment protocols are available and intraoperative monitoring was not possible, so far. Here we evaluate the potential benefits of the intraoperative assessment by microscope-integrated intraoperative optical coherence tomography (MI-OCT) of corneal optical properties during PTK. METHODS: Retrospective study of eight patients (one male, seven females; age range, 43-80 years, mean = 66.1 years) using an 840-nm microscope-integrated spectral-domain OCT (iOCT; OptoMedical Technologies, Luebeck, Germany adapted to HS Hi-R Neo 900A, Haag Streit Surgical, Wedel, Germany). Images were acquired before and after corneal abrasion and after PTK. For PTK, a SCHWIND Amaris 750S excimer laser (SCHWIND eye-tech-solutions GmbH und KO. KG) was used. Parameters assessed were the central corneal thickness (CCT), changes in central depth-dependent corneal tissue intensity (TI), and corneal surface roughness (SR) in cross-sectional images of the cornea. RESULTS: Intraoperative monitoring using microscope-integrated OCT was possible in all patients at all time points. TI of the anterior corneal stroma decreased significantly (p = 0.037) after PTK (T1 = 15.1 ± 3.6, T2 = 15.0 ± 3.84, T3 = 13.7 ± 3.38), but not after corneal abrasion alone, indicating increased transparency caused by excimer laser PTK. CCT was significantly lower after corneal abrasion (p = 0.017), but not after PTK (T1 = 630.4 ± 70 µm, T2 = 544.1 ± 59.4 µm, T3 = 558.3 ± 52.5 µm. SR significantly decreased (p = 0.043) after PTK (T1 = 614.4 ± 37.5 pixels, T2 = 634.4 ± 35.6 pixels, T3 = 611.0 ± 40.3 pixels). CONCLUSIONS: Intraoperative OCT allows real-time imaging during PTK and the assessment of corneal optical transparency and its surface roughness. It has to be clarified in larger studies if these parameters correlate with later postoperative visual outcomes.

Optimization Strategies for Bruch's Membrane Opening Minimum Rim Area Calculation: Sequential versus Simultaneous Minimization.

To compare a simultaneously optimized continuous minimum rim surface parameter between Bruch's membrane opening (BMO) and the internal limiting membrane to the standard sequential minimization used for calculating the BMO minimum rim area in spectral domain optical coherence tomography (SD-OCT). In this case-control, cross-sectional study, 704 eyes of 445 participants underwent SD-OCT of the optic nerve head (ONH), visual field testing, and clinical examination. Globally and clock-hour sector-wise optimized BMO-based minimum rim area was calculated independently. Outcome parameters included BMO-globally optimized minimum rim area (BMO-gMRA) and sector-wise optimized BMO-minimum rim area (BMO-MRA). BMO area was 1.89 ± 0.05 mm2. Mean global BMO-MRA was 0.97 ± 0.34 mm2, mean global BMO-gMRA was 1.01 ± 0.36 mm2. Both parameters correlated with r = 0.995 (P < 0.001); mean difference was 0.04 mm2 (P < 0.001). In all sectors, parameters differed by 3.0-4.2%. In receiver operating characteristics, the calculated area under the curve (AUC) to differentiate glaucoma was 0.873 for BMO-MRA, compared to 0.866 for BMO-gMRA (P = 0.004). Among ONH sectors, the temporal inferior location showed the highest AUC. Optimization strategies to calculate BMO-based minimum rim area led to significantly different results. Imposing an additional adjacency constraint within calculation of BMO-MRA does not improve diagnostic power. Global and temporal inferior BMO-MRA performed best in differentiating glaucoma patients.

Intraday Repeatability of Bruch's Membrane Opening-Based Neuroretinal Rim Measurements.

Purpose: To assess possible intraday variability in Bruch's membrane opening-based (BMO) assessment of neuroretinal rim by spectral-domain optical coherence tomography (SD-OCT) of the optic nerve head (ONH) as well as to evaluate its independence from intraindividual IOP changes. Methods: In this noninterventional, prospective cohort study, 51 consecutively enrolled patients with glaucoma or ocular hypertension underwent standardized SD-OCT of the ONH and IOP assessment at two different time points with a time gap of 5 hours on the same day. Random effects models, intraclass correlation coefficients (ICC) and Bland-Altman plots were used to analyze repeatability of BMO minimum rim width (BMO-MRW) and area (BMO-MRA) and peripapillary retinal nerve fiber layer (RNFL) thickness measurements. Results: Mean BMO area was 1.86 ± 0.30 mm2. At baseline, mean BMO-MRW was 206.46 ± 0.86 µm, mean BMO-MRA was 0.89 ± 0.34 mm2, and mean RNFL thickness was 71.61 ± 0.20 µm. Within intraindividual measurements, mean difference was 2.95 µm or 1.76% for BMO-MRW, 0.02 mm2 or 2.68% for BMO-MRA, and 1.18 µm or 1.89% for RNFL thickness. Comparing time points, η2 was 0.001 (P = 0.757) for RNFL thickness, 0.043 (P = 0.01) for BMO-MRA, and 0.07 (P = 0.06) for BMO-MRW. Intraclass correlation coefficients were greater than 0.994, respectively. Variability of morphometric parameters did not correlate to intraindividual IOP fluctuations. Conclusions: Assessment of BMO-MRW by SD-OCT shows high intraday repeatability, which is comparable to the parameter RNFL thickness. Variability seems not to be driven by typical moderate intraday fluctuations of IOP. The two-dimensional parameter BMO-MRA incorporates a fairly higher intraindividual variability.

Corneal Densitometry for Quantification of Corneal Deposits in Monoclonal Gammopathies.

PURPOSE: To assess the capability of Scheimpflug-based densitometry of the cornea to quantify light chain deposits in patients with active monoclonal gammopathies. METHODS: This is a case-control study in which data from a leading tertiary university center in myeloma care were analyzed. Ten eyes of 5 patients with monoclonal gammopathy and 26 eyes of 13 healthy controls undergoing clinical evaluation and Scheimpflug-based measurements were included in the study. The main outcome measures were densitometry data of the 4 corneal layers-anterior layer (AL), central layer (CL), posterior layer, and total layer (TL)-in 4 different annuli (central annular zone 0-2 mm, intermediate annular zone 2-6 mm, peripheral annular zone 6-10 mm, and total annular zone 0-12 mm). RESULTS: In 8 eyes of 4 patients with IgG-based gammopathy, corneal light backscatter was highest in the AL and decreased with increasing corneal depth. The peripheral annular zone showed a higher densitometry value compared with the corneal center. Compared with healthy controls, the AL (P < 0.001), the CL (P < 0.001), and the TL (P < 0.001) had significantly higher corneal light backscatter in patients with gammopathy in the total and the peripheral annular zones. In one patient with predominantly IgA-based disease, corneal light backscatter was not elevated. CONCLUSIONS: Scheimpflug-based densitometry of the cornea is able to quantify opacification by immunoglobulin G light chain deposits in monoclonal gammopathies. This noninvasive technique can complement presently used in vivo confocal microscopy and corneal photography to objectivize corneal changes. Densitometry might allow monitoring of corneal immunoglobulin deposits in follow-up examinations.

Neuroretinal rim in non-glaucomatous large optic nerve heads: a comparison of confocal scanning laser tomography and spectral domain optical coherence tomography.

BACKGROUND/AIMS: To compare margin-based rim area measurements from confocal scanning laser tomography (CSLT) with Bruch's membrane opening (BMO)-based measurements from spectral domain optical coherence tomography (SD-OCT) by analysis of a group of non-glaucomatous eyes with large optic discs, so-called macrodiscs (disc size >2.45 mm2 in CSLT). Objective is to create a reference base for large optic nerve heads in SD-OCT diagnostics. METHODS: 102 eyes received CSLT and SD-OCT measurements and clinical examination on the same day. Visual field testing confirmed absence of glaucomatous defects. Statistical and correlation analysis was performed for rim area by CSLT as well as retinal nerve fibre layer thickness (RNFLT) and BMO minimal rim width (BMO-MRW) by SD-OCT. RESULTS: Mean disc size in CSLT was 2.98±0.4 mm2 (range 2.45-4.23), mean rim area of 1.55±0.4 mm2. BMO area was 2.51±0.33 mm2 (range 1.61-3.51), mean global RNFLT was 79.55±17.2 µm, mean global BMO-MRW was 234.84±48.3 µm. Correlation of BMO-MRW to global RNFLT was stronger (r=0.60, p<10-5) than correlation of CSLT rim area to global RNFLT (r=0.26, p=0.24). BMO-MRW and CSLT rim area correlated with r=0.59 (p<10-5). BMO-MRW and RNFLT significantly decreased with increasing age (p<0.001). Annual loss of BMO-MRW was 0.8 µm/year (R2=0.14, p<0.001), loss of RNFLT was 0.27 µm/year (R2=0.17, p=0.001). CONCLUSIONS: In large optic discs, BMO-MRW is thinner compared with normal-sized discs and correlates better than CSLT parameters with the RNFLT. An age-depended loss of BMO-MRW needs to be taken into account in evaluation of the neuroretinal rim.

The use of Bruch's membrane opening-based optical coherence tomography of the optic nerve head for glaucoma detection in microdiscs.

PURPOSE: To assess the performance of Bruch's membrane opening (BMO)-based spectral domain optical coherence tomography (SD-OCT) of the optic nerve head for glaucoma detection in microdiscs in comparison with confocal scanning laser tomography (CSLT). DESIGN: Retrospective cohort study. METHODS: 82 eyes of 82 patients with disc size <1.63 mm2 underwent SD-OCT and CSLT measurements, visual field testing and clinical examination. BMO-based minimal rim width (BMO-MRW), retinal nerve fibre layer thickness (RNFLT) in SD-OCT and rim area measured in CSLT were compared and correlated with visual field defects. RESULTS: 51 patients with glaucoma, 11 patients with ocular hypertension (OHT) and 20 healthy controls had a mean disc area of 1.36±0.19 mm2 in CSLT, and BMO area was 1.45±0.22 mm2 (r=0.17; p=0.12). In patients with glaucoma, visual field mean defect was -7.5±6.7 dB. Global BMO-MRW correlated better with visual field function (Spearman's r=0.65; p<0.001) than RNFLT (r=0.58; p≤0.001) and CSLT rim area (r=0.47; p=0.004). BMO-MRW significantly deteriorated with progressive visual field loss (p<0.001). In receiver operating characteristic analysis, sensitivity of BMO-MRW was 68.6% at 95% specificity (area under curve (AUC)=0.87), similar to sensitivity of RNFLT (66.4%; AUC=0.81). Performance of CSLT rim area was significantly worse (AUC=0.70, p=0.008). In healthy controls, mean BMO-MRW was 344.3±64.1 µm, mean RNFLT 78.0±11.3 µm and CSLT mean rim area 1.07±0.18 mm2. CONCLUSIONS: In small optic discs, BMO-MRW and peripapillary RNFLT (OCT) have similar sensitivity to discriminate patients with glaucoma from normal controls; both exceed CSLT rim area in diagnostic power. In glaucomatous patients, BMO-MRW correlates strongest with visual field function.

Novel Bruch's Membrane Opening Minimum Rim Area Equalizes Disc Size Dependency and Offers High Diagnostic Power for Glaucoma.

Purpose: The purpose of this study was to assess the diagnostic power of the novel two-dimensional parameter Bruch's membrane opening minimal rim area (BMO-MRA) in spectral-domain optical coherence tomography (SD-OCT) for detection of glaucoma compared to minimal rim width (BMO-MRW) and retinal nerve fiber layer (RNFL) thickness in large and small optic discs. Methods: In this case-control, cross-sectional study, 207 eyes of 207 participants, including 89 controls and 97 patients with glaucoma and 21 with ocular hypertension (OHT), with a disc size <1.63 mm2 or >2.43 mm2 underwent SD-OCT, confocal laser scanning tomography (CSLT), visual field testing, and clinical examination. Bruch's membrane opening-MRA BMO-MRW, RNFL thickness of SD-OCT and disc margin rim area (DM-RA) of CSLT were evaluated and analyzed for diagnostic power to detect glaucoma. Results: In healthy eyes with macrodiscs, mean BMO-MRW of 243.14 ± 43.12 µm was significantly smaller than BMO-MRW in microdiscs (338.97 ± 69.39; P < 0.001). Bruch's membrane opening-MRA was comparable between disc size groups with 1.22 ± 0.25 mm2 for macrodiscs and 1.26 ± 0.27 mm2 for microdiscs (P = 0.51), as was RNFL thickness (82.69 ± 15.76 µm versus 78.53 ± 11.01 µm, respectively; P = 0.28). Perimetric mean deviation was -8.7 ± 6.3 dB in glaucoma and -0.6 ± 1.60 dB in OHT patients. Correlation of BMO-MRA and visual field function was rho (ρ) = 0.70 (P < 0.001). Diagnostic power to differentiate glaucoma patients was highest for BMO-MRA. Partial area under the curve (pAUC) for BMO-MRA was 0.14 for specificity between 0.8 and 1.0, exceeding pAUCs of BMO-MRW (P < 0.001), RNFL thickness (P = 0.03), and DM-RA (P = 0.01). Conclusions: Bruch's membrane opening-based minimum rim area measurements offer advantages compared to one-dimensional parameters assessing neuroretinal rim by SD-OCT. In nonglaucomatous eyes, BMO-MRA values seem comparable for the full range of disc sizes. Bruch's membrane opening-MRA surpasses other parameters in diagnostic power for glaucoma.

In Situ Corneal Cross-Linking for Recurrent Corneal Melting After Boston Type 1 Keratoprosthesis.

PURPOSE: To present a new treatment modality for recurrent corneal melting in a patient with a Boston type I keratoprosthesis (B-KPro) including in situ corneal cross-linking (CXL) and lamellar keratoplasty (LKP) as combined treatment. METHODS: Case report. RESULTS: Our report concerns a 27-year-old man whose case history involved a severe chemical burn of his left eye. After failed penetrating keratoplasty and limbal stem cell transplantation, the patient underwent B-KPro implantation. Starting 1 month after surgery, recurrent corneal melting around the B-KPro developed, which was eventually treated by combining LKP, amniotic membrane transplantation, and in situ CXL. Optical coherence tomography imaging and follow-up for 12 months showed stable corneal healing without new melting or erosion. The ultraviolet A treatment did not seem to damage the material of the B-KPro. CONCLUSIONS: In situ CXL using riboflavin and ultraviolet A light combined with LKP and amniotic membrane transplantation can be an effective management option to treat recurrent corneal melting after B-KPro implantation.

Long-term alterations of systemic vascular endothelial growth factor levels in patients treated with ranibizumab for age-related macular degeneration.

PURPOSE: To analyze long-term changes of systemic vascular endothelial growth factor (VEGF) levels in patients treated with ranibizumab for neovascular age-related macular degeneration. METHODS: Sixty-one patients with neovascular age-related macular degeneration and 68 age-matched controls were included in the study. Patients were treated with ranibizumab on a pro re nata regimen. Plasma samples were collected before initiation of treatment and after 1 year (30 patients) or 2 years (31 patients) of treatment. Vascular endothelial growth factor was measured by Luminex microbead analysis. RESULTS: At baseline, patients with neovascular age-related macular degeneration and controls did not differ significantly in VEGF levels (P = 0.062). There was a significant decline in systemic VEGF levels of 39.5% after 1 year (34.2 ± 17.2 pg/mL to 20.7 ± 14.0 pg/mL; P = 7.50 × 10(-5)) and of 46.7% after 2 years (40.4 ± 24.1 pg/mL to 21.5 ± 23.3 pg/mL; P = 2.48 × 10(-4)) of treatment. Patients with persistent activity of choroidal neovascularization showed a significantly smaller decrease of plasma VEGF levels than patients with dry intervals despite the higher number of injections (P = 0.048). CONCLUSION: In addition to immediate effects limited to days if not hours, ranibizumab also leads to long-term alterations of systemic VEGF to subnormal levels. Patients with persistent choroidal neovascularization activity showed a less pronounced VEGF decrease. Therefore, VEGF levels might be a useful marker for treatment response.

Upregulation of TGF-ß1 in experimental proliferative vitreoretinopathy is accompanied by epithelial to mesenchymal transition.

BACKGROUND: Proliferative vitreoretinopathy (PVR) is characterized by epithelial to mesenchymal transition (EMT) of retinal pigment epithelium (RPE) cells and consecutive formation of fibrous membranes, leading to retinal redetachment. Transforming growth factor beta (TGF-ß) has been suggested to play an important role in this process, but the role of TGF-ß isoforms is unknown. METHODS: In pigmented rabbits (n = 14), PVR was induced by cryopexy and a full-thickness limbus-parallel incision. PVR was evaluated by indirect ophthalmoscopy. Concentrations of TGF-ß isoforms were determined by multiplex bead assay analysis in aqueous humor (AH) and vitreous samples. EMT marker vimentin was analyzed by western blot. Masson's-trichrome, haematoxilin and eosine (H&E), and immunohistochemical analysis for EMT marker alpha SMA were performed on cross-sections of eyes. RESULTS: PVR was induced in all treated eyes. The number of quadrants affected by PVR was 1 (n = 5), 2 (n = 2), 3 (n = 2), 4 (n = 5). Vimentin and alpha SMA were expressed during PVR development. During PVR development, both TGF-ß1 levels (AH: p = 0.001; vitreous: p = 0.002) and TGF-ß2 levels increased (AH: p = 0.027; vitreous: p = 0.02), while TGF-ß3 was not detected at any timepoint. The increase was more pronounced for TGF-ß1 than for TGF- ß2 (AH: p = 0.002; vitreous: p = 0.0005), and only TGF-ß1 correlated with the amount of PVR (p = 0.024, r = 0,723). CONCLUSIONS: Development of PVR membranes was accompanied by a pronounced upregulation of TGF-ß1, rather than TGF-ß2. Therefore TGF-ß1 could be a promising target for inhibition of PVR.

Polymorphisms in vascular endothelial growth factor receptor 2 are associated with better response rates to ranibizumab treatment in age-related macular degeneration.

PURPOSE: Intravitreal anti-vascular endothelial growth factor (VEGF) injections are currently the standard treatment for neovascular age-related macular degeneration (AMD), but a broad range of response rates has been observed. We evaluated the association of single nucleotide polymorphisms (SNPs) in VEGF genes and their receptors (VEGFR) with the response rate to ranibizumab in 366 patients with neovascular AMD. DESIGN: Case series study. PARTICIPANTS: A total of 366 eyes of 366 patients with neovascular AMD. METHODS: Visual acuity (VA) was determined at baseline, after 3 monthly ranibizumab injections, and after 1 year of treatment. Genotyping of 126 SNPs in the genes encoding VEGF family members VEGFA, VEGFB, VEGFC, VEGFD (FIGF), and placental growth factor (PGF); VEGF receptors VEGFR1 (FLT1), VEGFR2 (KDR), and VEGFR3 (FLT4); and the gene encoding pigment epithelium-derived factor (PEDF) (SERPINF1) was performed. MAIN OUTCOME MEASURES: The changes in VA after 3 injections and after 1 year of treatment and their association with VEGF and VEGFR genotypes. RESULTS: Univariate analyses of variance (ANOVAs) revealed a significant effect of SNP rs4576072 in the VEGFR2 gene on VA change after 12 months (F[1,235] = 14.05; P = 0.02). A stepwise linear regression analysis returned a model (P = 0.01) with SNPs rs4576072 and rs6828477 in the VEGFR2 gene as independent predictors for VA change after 12 months, with a mean increase in VA of 0.26 on the logarithm of the minimum angle of resolution (logMAR) scale in patients with 3 contributing minor alleles compared with a loss of 0.03 logMAR in patients with no minor allele. CONCLUSIONS: Polymorphisms in the VEGFR2/KDR gene significantly influence visual outcome in patients receiving ranibizumab treatment for neovascular AMD. This study shows that genetic variation partially explains the wide range of response to ranibizumab treatment, which in the future might help clinicians tailoring medical interventions to individual needs.