Seth M. Pantanelli

Glucocorticoid-induced tumor necrosis factor receptor negatively regulates activation of human primary natural killer (NK) cells by blocking proliferative signals and increasing NK cell apoptosis.

Glucocorticoid-induced tumor necrosis factor receptor (GITR), found constitutively expressed on human primary natural killer (NK) cells at low levels was up-regulated upon stimulation by either Toll-like receptor ligand or NK cell growth factor, interleukin (IL)-15. cDNA microarray analysis showed that engagement of GITR primarily suppressed the activation of NF-KB pathway of NK cells and up-regulated anti-inflammatory genes heme oxygenase-1 and IL-10. Further analysis revealed that GITR activation suppressed NK cell proliferation in response to IL-15. GITR activation also suppressed proinflammatory cytokine secretion and increased NK cell apoptosis. GITR activation resulted in blocked phosphorylation of Stat5 and Akt, which may have contributed to the observed antiproliferative effect of GITR on NK cells. Increased apoptosis was independent of the Fas-FasL pathway, but Bcl-XL and phospho-Bad protein expressions were diminished, suggesting involvement of the mitochondrial apoptosis pathway. The results suggest that although GITR is an activation marker for NK cells similar to that for T cells, GITR serves as a negative regulator for NK cell activation. Our studies demonstrate a novel physiological role of GITR on NK cells.

CD4+Foxp3+ T-Regulatory Cells in Noninfectious Uveitis

OBJECTIVE To evaluate CD4(+)Foxp3(+) (forkhead box P3) T-regulatory cell populations in patients with uveitis and to determine if T-regulatory cell populations are associated with disease features. METHODS Patients with uveitis were evaluated for CD4(+)Foxp3(+) T-regulatory cells by flow cytometry. Systemic and ocular diagnoses, disease activity, and the presence of cystoid macular edema were reviewed. Percentages of CD4(+)Foxp3(+) lymphocytes were compared for patients with inactive vs active disease, systemic vs ocular diagnoses, and the presence or absence of cystoid macular edema. Real-time polymerase chain reaction testing was performed on 2 patients with extremely low CD4(+)Foxp3(+) cell populations to assess Foxp3 mRNA. RESULTS A total of 20 patients with intermediate uveitis, posterior uveitis, and panuveitis were evaluated. The mean age was 40.6 years and the mean visual acuity was 20/57. Percentages of CD4(+)Foxp3(+) cells were lower in patients with active compared with inactive uveitis (P< .05). No differences in T-regulatory cells were observed between the other subgroups. Two patients with recalcitrant uveitis who demonstrated less than 1% CD4(+)Foxp3(+) lymphocytes showed extremely low or absent Foxp3 mRNA. CONCLUSION T-regulatory cells are reduced in patients with active compared with inactive disease. Severe depletion of CD4(+)Foxp3(+) T cells and Foxp3 mRNA in 2 patients with severe uveitis suggests that loss of the T-regulatory cells of uveitis may be a salient feature in certain patients.

Large-dynamic-range Shack-Hartmann wavefront sensor for highly aberrated eyes.

A conventional Shack-Hartmann wavefront sensor has a limitation that increasing the dynamic range usually requires sacrificing measurement sensitivity. The prototype large-dynamic-range Shack-Hartmann wavefront sensor presented resolves this problem by using a translatable plate with subapertures placed in conjugate with the lenslet array. Each subaperture is the same size as a lenslet and they are arranged so that they overlap every other lenslet position. Three translations of the plate are required to acquire four images to complete one measurement. This method increases the dynamic range by a factor of two with no subsequent change in measurement sensitivity and sampling resolution of the aberration. The feasibility of the sensor was demonstrated by measuring the higher order aberrations of a custom-made phase plate and human eyes with and without the plate.

Post-DSAEK optical changes: a comprehensive prospective analysis on the role of ocular wavefront aberrations, haze, and corneal thickness.

Purpose: The aim was to assess the visual impact of ocular wavefront aberrations, corneal thickness, and corneal light scatter prospectively after performing a Descemet stripping automated endothelial keratoplasty (DSAEK) in humans. Methods: Data were obtained prospectively from 20 eyes preoperatively and at 1, 3, 6, and 12 months post-DSAEK. At each visit, the best spectacle-corrected visual acuity and visual acuity with glare (brightness acuity testing) were recorded, and ocular wavefront measurements and corneal optical coherence tomography (OCT) were performed. The magnitude and the sign of individual Zernike terms [higher-order aberrations (HOAs)] were determined. Epithelial, host stromal, donor stromal, and total corneal thicknesses were quantified. The brightness and intensity profiles of OCT images were generated to quantify light scatter in the whole cornea, subepithelial region, anterior and posterior host stroma, interface, and donor stroma. Results: The mean best spectacle-corrected visual acuity and glare disability at low light levels improved from 1 to 12 months post-DSAEK. All corneal thicknesses and ocular lower-order aberrations and HOAs were found to be stable from 1 to 12 months, whereas total corneal, host stromal, and interface brightness intensities decreased significantly over the same period. A repeated measures analysis of variance performed across the follow-up period revealed that the change in scatter, but not the change in the HOAs, could account for the variability occurring in the acuity from 1 to 12 months post-DSAEK. Conclusions: Although ocular HOAs and scatter are both elevated over normal values post-DSAEK, our results demonstrate that the improvements in visual performance occurring over the first year post-DSAEK are associated with decreasing light scatter. In contrast, there were no significant changes in the ocular HOAs during this time. Because corneal light scatter decreased between 1 and 12 months despite there being stable corneal thicknesses over the same period, we conclude that factors that induced light scatter, other than tissue thickness or swelling (corneal edema), significantly impacted the visual improvements that occurred over time post-DSAEK. A better understanding of the cellular and extracellular matrix changes of the subepithelial region and interface, incurred by the surgical creation of a lamellar host–graft interface, and the subsequent healing of these tissues, is warranted.

Visual Performance with Wave Aberration Correction after Penetrating, Deep Anterior Lamellar, or Endothelial Keratoplasty

PURPOSE To investigate the contribution ocular aberrations have on visual performance by quantifying improvements in best-corrected visual acuity (VA) and contrast sensitivity (CS) obtained with higher-order aberration (HOA) correction after penetrating (PK), deep anterior lamellar (DALK), or Descemets stripping automated endothelial keratoplasty (DSAEK). METHODS Sixteen eyes were evaluated from 14 subjects who underwent PK (n = 5), DALK (n = 6), or DSAEK (n = 5) greater than 1 year prior to study enrollment. Ocular aberrations were measured and an adaptive optics system was used to correct ocular lower-order aberration (LOA) and HOA. VA and CS were measured for each subject with LOA or full-aberration correction. CS was measured at each of three spatial frequencies: 4, 8, and 12 cycles/deg. RESULTS All keratoplasty groups had more aberration than that of a normal myopic population and experienced significant VA gains with full-aberration correction (P < 0.0013). PK subjects had better VA than that of DSAEK subjects with LOA correction (logMAR VA 0.03 ± 0.05 vs. 0.25 ± 0.05; P = 0.0870). After HOA correction this trend persisted (P = 0.1734). DSAEK subjects also experienced less VA benefit from full-aberration correction than that of PK and DALK subjects. All keratoplasty groups demonstrated similar CS benefits from full-aberration correction despite differing higher-order root-mean-square magnitudes. CONCLUSIONS PK eyes had better logMAR VA than that of DSAEK eyes with LOA correction, whereas DALK eyes performed intermediate between the two. When full correction was applied, the same trend persisted. The findings suggest that factors other than aberration contribute to decrements in VA with DSAEK compared with PK.

Differentiation of malignant B-lymphoma cells from normal and activated T-cell populations by their intrinsic autofluorescence.

Patients with active posterior and intermediate uveitis have inflammatory cells in their vitreous; those with primary intraocular lymphoma have malignant B-lymphoma cells concomitantly. These cell types cannot be distinguished clinically. The goal of this study was to investigate intrinsic autofluorescence as a noninvasive way of differentiating immune and lymphomatous cell populations. Human primary T cells were stimulated with or without anti-CD3 plus anti-CD28 stimulation. B-lymphoma cells (CA46) were cultured separately. Five experimental groups were prepared: unstimulated T cells, stimulated T cells, CA46 cells, and stimulated T cells mixed with CA46 cells at a ratio of 1:3 or mixed at a ratio of 3:1. Samples were excited with three wavelengths and imaged with a confocal microscope. For each condition, the autofluorescent emissions from the sample were measured. In separate experiments, T cells or CA46 cells were injected into the anterior chamber of a BALB/c mouse eye and autofluorescence was measured. Pure T-cell and lymphoma populations were clearly distinguishable based on autofluorescence intensity spectra. CA46 cells were the least fluorescent when excited with 351-nm light, but most fluorescent when excited with longer wavelengths like 488 nm. Mixed populations of T cells and CA46 cells had emission intensities that fell predictably in between those of the pure populations. An ex vivo study showed that CA46 cells could be detected based on their intrinsic autofluorescence. Our studies showed that normal activated and malignant lymphocyte populations can be distinguished based on their intrinsic autofluorescent properties. Future work with in vivo models may prove useful in facilitating the diagnosis of uveitis and other ocular diseases.

Quiescent posterior capsule trauma after intravitreal injection: Implications for the cataract surgeon

1. Vallance JH, Ahmed M, Dhillon B. Cataract surgery and consent: recall, anxiety, and attitude toward trainee surgeons preoperatively and postoperatively. J Cataract Refract Surg 2004; 30:1479–1485 2. Habib MS, Bunce CV, Fraser SG. The role of case mix in the relation of volume and outcome in phacoemulsification. Br J Ophthalmol 2005; 89:1143–1146. Available at: http://www. pubmedcentral.nih.gov/picrender.fcgi?artidZ1772833b 88:1242–1246. Available at: http://www. pubmedcentral.nih.gov/picrender.fcgi?artidZ1772356b 23:31–37. Available at: http://www.nature.com/ eye/journal/v23/n1/pdf/6703049a.pdf. Accessed February 7, 2011

Anterior capsulotomy outcomes: A comparison between two femtosecond laser cataract surgery platforms

PURPOSE To compare capsulotomy outcomes between two femtosecond laser platforms for cataract surgery. METHODS This retrospective case series included 205 eyes of 162 consecutive patients (75 male and 87 female) aged 68.32 ± 10.27 years (range: 38 to 92 years) who underwent femtosecond laser-assisted cataract surgery (FLACS) using either the LenSx (Alcon Laboratories, Inc., Fort Worth, TX) or the Catalys (Abbott Medical Optics, Abbott Park, IL) laser platform. Intraoperative assessment of the capsulotomy was performed and graded as one of the following four types: complete treatment pattern (type 1), microadhesions (type 2), incomplete treatment pattern (type 3), and complete pattern but not continuous (type 4). RESULTS Ninety-seven eyes underwent FLACS using the LenSx platform and 108 eyes using the Catalys platform. Ninety-four capsulotomies in the LenSx group were graded as type 1 (96.91%), and three were graded as type 2 (3.09%); all capsulotomies in the Catalys group were graded as type 1 (100%). There was no statistically significant difference between the LenSx and Catalys groups (P > .05). No intraoperative complications (specifically, no posterior capsular extension or vitreous loss) were noticed in any of the eyes included in the study. CONCLUSIONS The LenSx and Catalys laser platforms demonstrate similar capsulotomy outcomes. The microadhesions that were demonstrated in three cases in the LenSx group did not lead to intraoperative or postoperative complications.

Trends in cataract surgery training curricula

PURPOSE To evaluate trends in cataract surgery training curricula and factors affecting timing of resident participation as a primary surgeon. SETTING Pennsylvania State College of Medicine, Hershey, Pennsylvania, USA. DESIGN Cross-sectional study of anonymous survey results. METHODS A description of the study and link to an online survey was e-mailed to program directors of each ophthalmology residency training program accredited by the Accreditation Council for Graduate Medical Education (ACGME). RESULTS Fifty-one (44%) of the 116 program directors completed the survey. First-year, second-year, and third-year residents performed a mean of 2, 25, and 155 phacoemulsification surgeries, respectively, as a primary surgeon. Only 1 program (2%) required residents to perform extracapsular cataract extraction (ECCE) before performing phacoemulsification. Clear corneal phacoemulsification was the first technique taught to trainees at 91% of programs. More than two thirds (71%) of program directors indicated that their program had a cataract surgery training curriculum designed to transition residents gradually to the operating room. These curricula included structured wet laboratory (92%) and lecture (89%) components. Inadequate resident knowledge and surgical skill base (57%) and anticipation of increased surgical complication risk (37%) were the most commonly reported factors impeding earlier exposure to phacoemulsification in residency. CONCLUSIONS Results show that residents today begin surgical training with phacoemulsification rather than ECCE, perform a higher number of phacoemulsification surgeries than is required by the ACGME, and begin performing phacoemulsification as early as their first or second year of residency. Despite these evolutions, 29% of respondent ACGME-accredited ophthalmology residency programs reported not having a formal cataract surgery training curriculum.

Retinal manifestations of autoimmune and inflammatory disease.

Autoimmune or inflammatory diseases may affect multiple organs throughout the body, but in fact often have very specific targets (ie, arteries and veins, melanocytes, or the nervous system). As the retina is the most metabolically active tissue in the body, its dependence on the circulatory system makes it particularly susceptible to any process resulting in vasculitis. Similarly, presence of melanin in the retinal pigment epithelium (RPE) exposes the retina to those diseases that involve melanocytes elsewhere in the body. Finally, mere presence of the nerve fiber layer and optic nerve makes the retina susceptible to those diseases that target the nervous system. Therefore, it is not surprising that the retina is a frequent location for manifestation of these disorders; a careful examination not only sheds light on the pathologic process, but often allows one to predict active versus quiescent disease activity elsewhere in the body. The purpose of this study is to review the epidemiology, pathogenesis, diagnostic criteria, and retinal manifestations for a number of autoimmune or inflammatory-mediated diseases.