Eyelid Reconstruction: Pediculated versus Non-Pediculated.

BACKGROUND: To compare the functional and cosmetic outcome of pediculated versus free anterior and posterior lamella reconstruction after large eyelid defects due to malignancy excision. PATIENTS AND METHODS: A retrospective study over 2 years with 12 patients matching the criterion of pediculated versus non-pediculated transplants out of a cohort of 124 tumor excisions. The mean age was 76 ± 8 years of the 7 male and 5 female patients. In the majority of cases, more than half of the eyelid was excised. The posterior lamella was always reconstructed with tarsal tissue, and the anterior lamella mostly with an upper eyelid skin graft. The postoperative follow-up time was between 2 months and 1 year. RESULTS: Nodular basal cell carcinoma was the prevailing histology (6 patients). The reconstruction techniques included a Hughes procedure (four patients) or a free tarsal graft with a pediculated skin flap (four patients), respectively. In the remaining four patients, a combination of pediculated/free anterior AND posterior lamellae was performed. Four patients had a one-stage and eight patients a 2nd stage procedure with a mean time until tarsoconjunctival flap reopening of 16 ± 2 days. CONCLUSIONS: No difference was found in the final functional outcome in pediculted versus free grafts. The cosmetic result was better in anterior lamella reconstructions with a pediculated flap, which usually allows a one-stage procedure. The 2nd stage procedure could be performed after 2 weeks without any complications.

Long-term Follow-up of Canaliculotomies after Canaliculitis.

BACKGROUND: Canaliculitis is often misdiagnosed. There are several conservative and surgical treatment options. PATIENTS AND METHODS: Retrospective analysis of 14 canaliculotomies in 10 patients with canaliculitis. The overall length of the surgically induced opening was measured and compared to the corresponding untreated lacrimal punctae. Lacrimal duct concrements were liberated and sent for microbiological and histological analysis. Patient satisfaction and relief of symptoms were documented as well as clinical findings. RESULTS: Mean age was 59 ± 10 years (36 - 73 years) with balanced gender distribution. Mean follow-up time was 13 ± 8 months (4 - 27 months). Canaliculotomy was performed on 12 out of 14 inflamed canaliculi; in 2 cases, 3-snip punctoplasty was sufficient. The surgically induced length of the cuts was 1.7 ± 0.9 mm (0.4 - 3.7 mm). In 13 out of 14 cases, macroscopic concrements were found intraoperatively and actinomyces was verified histologically. Nine patients were free of symptoms postoperatively, and one patient manifested markedly less epiphora. CONCLUSIONS: The canaliculi remained open within the long-term follow-up period without any drawbacks to the lacrimal outflow. No recurrent infections were seen.

Molecular imaging reveals elevated VEGFR-2 expression in retinal capillaries in diabetes: a novel biomarker for early diagnosis.

Diabetic retinopathy (DR) is a microvascular complication of diabetes and a leading cause of vision loss. Biomarkers and methods for early diagnosis of DR are urgently needed. Using a new molecular imaging approach, we show up to 94% higher accumulation of custom designed imaging probes against vascular endothelial growth factor receptor 2 (VEGFR-2) in retinal and choroidal vessels of diabetic animals (P<0.01), compared to normal controls. More than 80% of the VEGFR-2 in the diabetic retina was in the capillaries, compared to 47% in normal controls (P<0.01). Angiography in rabbit retinas revealed microvascular capillaries to be the location for VEGF-A-induced leakage, as expressed by significantly higher rate of fluorophore spreading with VEGF-A injection when compared to vehicle control (26±2 vs. 3±1 µm/s, P<0.05). Immunohistochemistry showed VEGFR-2 expression in capillaries of diabetic animals but not in normal controls. Macular vessels from diabetic patients (n=7) showed significantly more VEGFR-2 compared to nondiabetic controls (n=5) or peripheral retinal regions of the same retinas (P<0.01 in both cases). Here we introduce a new approach for early diagnosis of DR and VEGFR-2 as a molecular marker. VEGFR-2 could become a key diagnostic target, one that might help to prevent retinal vascular leakage and proliferation in diabetic patients.

Noninvasive molecular imaging reveals role of PAF in leukocyte-endothelial interaction in LPS-induced ocular vascular injury.

Uveitis is a systemic immune disease and a common cause of blindness. The eye is an ideal organ for light-based imaging of molecular events underlying vascular and immune diseases. The phospholipid platelet-activating factor (PAF) is an important mediator of inflammation, the action of which in endothelial and immune cells in vivo is not well understood. The purpose of this study was to investigate the role of PAF in endothelial injury in uveitis. Here, we use our recently introduced in vivo molecular imaging approach in combination with the PAF inhibitors WEB 2086 (WEB) and ginkgolide B (GB). The differential inhibitory effects of WEB and GB in reducing LPS-induced endothelial injury in the choroid indicate an important role for PAF-like lipids, which might not require the PAF receptor for their signaling. P-selectin glycoprotein ligand-1-mediated rolling of mouse leukocytes on immobilized P-selectin in our autoperfused microflow chamber assay revealed a significant reduction in rolling velocity on the cells' contact with PAF. Rolling cells that came in contact with PAF rapidly assumed morphological signs of cell activation, indicating that activation during rolling does not require integrins. Our results show a key role for PAF in mediating endothelial and leukocyte activation in acute ocular inflammation. Our in vivo molecular imaging provides a detailed view of cellular and molecular events in the complex physiological setting.

An animal model of spontaneous metabolic syndrome: Nile grass rat.

Metabolic syndrome (MetS) is a prevalent and complex disease, characterized by the variable coexistence of obesity, dyslipidemia, hyperinsulinaemia, and hypertension. The alarming rise in the prevalence of metabolic disorders makes it imperative to innovate preventive or therapeutic measures for MetS and its complications. However, the elucidation of the pathogenesis of MetS has been hampered by the lack of realistic models. For example, the existing animal models of MetS, i.e., genetically engineered rodents, imitate certain aspects of the disease, while lacking other important components. Defining the natural course of MetS in a spontaneous animal model of the disease would be desirable. Here, we introduce the Nile grass rat (NGR), Arvicanthis niloticus, as a novel model of MetS. Studies of over 1100 NGRs in captivity, fed normal chow, revealed that most of these animals spontaneously develop dyslipidemia (P<0.01), and hyperglycemia (P<0.01) by 1 yr of age. Further characterization showed that the diabetic rats develop liver steatosis, abdominal fat accumulation, nephropathy, atrophy of pancreatic islets of Langerhans, fatty streaks in the aorta, and hypertension (P<0.01). Diabetic NGRs in the early phase of the disease develop hyperinsulinemia, and show a strong inverse correlation between plasma adiponectin and HbA1c levels (P<0.01). These data indicate that the NGR is a valuable, spontaneous model for exploring the etiology and pathophysiology of MetS as well as its various complications.

Atrial natriuretic peptide reduces vascular leakage and choroidal neovascularization.

Atrial natriuretic peptide (ANP) is a hormone with diuretic, natriuretic, and vasodilatory properties. ANP blocks vascular endothelial growth factor (VEGF) production and signaling in vitro; however, its role in vascular leakage and angiogenesis is unknown. In vitro, retinal barrier permeability (transepithelial electrical resistance (TEER)) was measured in cultured retinal endothelial (HuREC) and retinal epithelial (ARPE-19) cells with VEGF (10 ng/ml), ANP (1 pM to 1 micromol/L), and/or isatin, an ANP receptor antagonist. In vivo, blood-retinal barrier (BRB) leakage was studied using the Evans Blue dye technique in rats treated with intravitreal injections of ANP, VEGF, or vehicle. Choroidal neovascularization was generated by laser injury, and 7 days later, lesion size and leakage was quantitated. ANP significantly reversed VEGF-induced BRB TEER reduction in both HuREC and ARPE-19 cells, modeling the inner and the outer BRB, respectively. Isatin, a specific ANP receptor antagonist, reversed ANP's effect. ANP reduced the response of ARPE-19 cells to VEGF apically but not basolaterally, suggesting polarized expression of the ANP receptors in these cells. ANP's TEER response was concentration but not time dependent. In vivo, ANP significantly reduced VEGF-induced BRB leakage and the size of laser-induced choroidal neovascularization lesions. In sum, ANP is an effective inhibitor of VEGF-induced vascular leakage and angiogenesis in vivo. These results may lead to new treatments for ocular diseases where VEGF plays a central role, such as age-related macular degeneration or diabetic retinopathy.

In vivo imaging of endothelial injury in choriocapillaris during endotoxin-induced uveitis.

Early detection of ocular inflammation may prevent the occurrence of structural damage or vision loss. Here, we introduce a novel noninvasive technique for molecular imaging and quantitative evaluation of endothelial injury in the choriocapillaris of live animals, which detects disease earlier than currently possible. Using an established model of ocular inflammation, endotoxin-induced uveitis (EIU), we visualized the rolling and adhesive interaction of fluorescent microspheres conjugated to recombinant P-selectin glycoprotein ligand-Ig (rPSGL-Ig) in choriocapillaris using a scanning laser ophthalmoscope (SLO). The number of rolling microspheres in the choriocapillaris peaked 4-10 h after LPS injection. The number of the accumulated microspheres peaked 4 h after LPS injection in the temporal choriocapillaris and 4 and 36 h after LPS injection in the central areas around the optic disk. Furthermore, we semiquantified the levels of P-selectin mRNA expression in the choroidal vessels by reverse transcription-PCR and found its pattern to match the functional microsphere interactions, with a peak at 4 h after LPS injection. These results indicate that PSGL-1-conjugated fluorescent microspheres allow specific detection of endothelial P-selectin expression in vivo and noninvasive assessment of endothelial injury. This technique may help to diagnose subclinical signs of ocular inflammatory diseases.

Characterization of azurocidin as a permeability factor in the retina: involvement in VEGF-induced and early diabetic blood-retinal barrier breakdown.

PURPOSE: Azurocidin, released by neutrophils during leukocyte-endothelial interaction, is a main cause of neutrophil-evoked vascular leakage. Its role in the retina, however, is unknown. METHODS: Brown Norway rats received intravitreal injections of azurocidin and vehicle control. Blood-retinal barrier (BRB) breakdown was quantified using the Evans blue (EB) dye technique 1, 3, and 24 hours after intravitreal injection. To block azurocidin, aprotinin was injected intravenously before the intravitreal injections. To investigate whether azurocidin plays a role in vascular endothelial growth factor (VEGF)-induced BRB breakdown, rats were treated intravenously with aprotinin, followed by intravitreal injection of VEGF(164). BRB breakdown was quantified 24 hours later. To investigate whether azurocidin may mediate BRB breakdown in early diabetes, aprotinin or vehicle was injected intravenously each day for 2 weeks to streptozotocin-induced diabetic rats, and BRB breakdown was quantified. RESULTS: Intravitreal injection of azurocidin (20 microg) induced a 6.8-fold increase in vascular permeability compared with control at 1-3 hours (P < 0.05), a 2.7-fold increase at 3 to 5 hours (P < 0.01), and a 1.7-fold increase at 24 hours (P < 0.05). Aprotinin inhibited azurocidin-induced BRB breakdown by more than 95% (P < 0.05). Furthermore, treatment with aprotinin significantly suppressed VEGF-induced BRB breakdown by 93% (P < 0.05) and BRB breakdown in early experimental diabetes by 40.6% (P < 0.05). CONCLUSIONS: Azurocidin increases retinal vascular permeability and is effectively blocked by aprotinin. The inhibition of VEGF-induced and early diabetic BRB breakdown with aprotinin indicates that azurocidin may be an important mediator of leukocyte-dependent BRB breakdown secondary to VEGF. Azurocidin may become a new therapeutic target in the treatment of retinal vascular leakage, such as during diabetic retinopathy.

Type, severity, management and outcome of ocular and adnexal firework-related injuries: the Rotterdam experience.

PURPOSE: To study the type, severity, management and outcome of firework-related adnexal and ocular injuries during New Year's Eve festivities. METHODS: A retrospective analysis of 123 injured patients (143 eyes) treated at the Rotterdam Eye Hospital between 2009 and 2013. All ages were included and analysed according to age, gender, active participant or bystander, laterality, location, dimension and severity of injury. Outcome parameter was the final best-corrected visual acuity. RESULTS: The mean age was 22 ± 13 years with 87% males and 53% bystanders. 52% were ≤18 years. There was a higher number of female than male bystanders (63% versus 51%, p = 0.30). 50% of the eyes sustained mild, 13% moderate and 37% severe trauma. Adults suffered more from severe injuries compared to children (42% versus 31%). The most frequent intervention was gunpowder removal (20%), followed by traumatic cataract surgery (12%) and amniotic membrane grafting (8%). 76% of patients were followed over 1 year. At the end of follow-up, 88 (61.5%) eyes had recovered fully, while 55 (38.5%) eyes suffered from persistent complications with reduced vision ≤0.8 in 30% of injured eyes. 15 patients (12%, 10 adults, five children) were considered legally blind (vision ≤0.1). Three (2%) eyes were subject to evisceration. CONCLUSION: Every year, around New Year's Eve 30-45 victims were referred to the Rotterdam Eye Hospital; 50% sustained moderate-to-severe trauma. In severe firework injuries, patients required multiple treatments that may not prevent permanent blindness and/or functional/cosmetic disfigurement. The majority was bystander and younger than 18 years.

Correlation Between Dynamic Contour Tonometry, Uncorrected and Corrected Goldmann Applanation Tonometry, and Stage of Glaucoma.

Importance: Accurate determination of intraocular pressure (IOP) is crucial for the diagnosis and management of glaucoma. Objective clinical evaluation of the correction equations for Goldmann applanation tonometry (GAT) is lacking. Objectives: To investigate the difference between corrected and conventional GAT and Pascal dynamic contour tonometry (DCT) measurements, as well as the correlation between discordant IOP values and stage of glaucoma. Design, Setting, and Participants: This prospective cross-sectional case series was conducted at the Department of Ophthalmology, University Hospital Zurich, and Talacker Eye Center between July 1, 2011, and May 31, 2016, among 112 white patients with glaucoma. Interventions: Intraocular pressure measurements were performed with GAT and DCT in a randomized order. Goldmann applanation tonometry measurements were modified with 5 correction equations. Main Outcomes and Measures: The primary end point was degree of concordance between corrected or uncorrected GAT and DCT measurements. The secondary end point was association between discordant IOP measurements and the stage of glaucoma, as assessed by the Glaucoma Severity Score. Results: Among the 112 patients (67 women and 45 men; mean [SD] age, 66.3 [13.1] years), 63 of the eyes in the study (56.3%) were left eyes and 85 patients (75.9%) were taking ocular antihypertensive medications. Mean (SD) IOP was 20.3 (4.5) mm Hg (95% CI, 19.4-21.1) as measured by DCT and 17.0 [4.1] mm Hg (95% CI, 16.3-17.8) as measured by GAT. The mean (SD) discordance between DCT and GAT measurements was -3.3 (2.0) mm Hg (95% CI, 2.9-3.6). The 5 corrected GAT values ranged from -2.7 to -5.4 mm Hg compared with DCT. The mean (SD) result of the Dresdner correction formula (17.6 [4.1] mm Hg) was closer to the DCT measurement than the original GAT measurement. The mean (SD) Glaucoma Severity Score was 4.7 (3.4) (95% CI, 4.1-5.4). The uncorrected discordance IOPDCT - IOPGAT showed a positive correlation with the Glaucoma Severity Score (rs = 0.33; P < .001) and a negative correlation with central corneal thickness (rs = -0.22; P = .02). Conclusions and Relevance: In comparison with DCT measurements, these data suggest that GAT values are significantly discordant in eyes with thin corneas and advanced glaucoma. Application of GAT-based correction formulas involves a possible risk of creating an even greater number of unpredictable measurement errors. Hence, we advise with caution, especially pertaining to eyes with thin corneas, to not place reliance on GAT readings, and abandon any correction formula. Trial Registration: clinicaltrials.gov Identifier: NCT01474070.

Molecular Imaging of Retinal Endothelial Injury in Diabetic Animals.

PURPOSE: Diabetic retinopathy is a leading cause of vision loss. There is a great need for early diagnosis prior to the occurrence of irreversible structural damages. Expression of endothelial adhesion molecules is observed before the onset of diabetic vascular damage; however, to date, these molecules cannot be visualized in vivo. METHODS: To quantify the expression of endothelial surface molecules, we generated imaging probes that bind to ICAM-1. The α-ICAM-1 probes were characterized via flow cytometry under microfluidic conditions. Probes were systemically injected into normal and diabetic rats, and their adhesion in the retinal microvessels was visualized via confocal scanning laser ophthalmoscopy. Histology was performed to validate in vivo imaging results. Vascular pathologies were visualized using trypsin-digested retinal preparations. RESULTS: The α-ICAM-1 probes showed significantly higher adhesion to retinal microvessels in diabetic rats than in normal controls (P < 0.01), whereas binding of control probes did not differ between the two groups. Western blotting results showed higher ICAM-1 expression in retinas of T1D animals than in normal controls. Retinal endothelial ICAM-1 expression was observed via molecular imaging before markers of structural damage, such as pericyte ghosts and acellular capillaries. CONCLUSION: Results indicate that molecular imaging can be used to detect subtle changes in the diabetic retina prior to the occurrence of irreversible pathology. Thus, ICAM-1 could serve as a diagnostic target in patients with diabetes. This study provides a proof of principle for non-invasive subclinical diagnosis in experimental diabetic retinopathy. Further development of this technology could improve management of diabetic complications.

High myopia as a risk factor for post-LASIK ectasia: a case report.

PURPOSE: To describe the case of a patient developing corneal ectasia following LASIK for the correction of myopic astigmatism. MATERIALS AND METHODS: A 39-year-old man underwent bilateral uneventful LASIK for myopic astigmatism of -10.25 -1.75 ×040 OD and -8.00 -2.50 ×005 OS. Preoperative corneal pachymetry was 542 micrometers OD and 543 micrometers OS. Preoperative corneal topography showed bilateral oblique bow-tie patterns. Central keratometry measurements were 45.12 D @ 124 / 43.87 D @ 34 OD and 44.87 D @ 78 / 43.12 D @ 168 OS. Keratoconus or forme fruste keratoconus were not present preoperatively. RESULTS: The residual stromal bed was 314 micrometers OD and 295 micrometers OS. Increasing astigmatism was documented progressively after LASIK. Central keratometry and topography were performed with evidence of ectasia OD at 17 months post-operatively and early evidence of ectasia OS at last follow-up of 58 months. CONCLUSION: High myopia appears to be a predisposing factor in this patient. High myopia may need to be considered as an ectasia risk factor independent of amount of ablation or residual stromal bed thickness and in the absence of forme fruste keratoconus. The possibility remains that ectasia was due to an unidentified risk factor or an intrinsic corneal problem with this patient's right eye.