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Purpose Exfoliation syndrome (XFS) is a condition characterized by the production of insoluble fibrillar aggregates (exfoliation material; XFM) in the eye and elsewhere. Many patients with XFS progress to exfoliation glaucoma (XFG), a significant cause of global blindness. We used quantitative mass spectrometry to analyze the composition of XFM in lens capsule specimens and in aqueous humor (AH) samples from patients with XFS, patients with XFG and unaffected individuals. Methods Pieces of lens capsule and samples of AH were obtained with consent from patients undergoing cataract surgery. Tryptic digests of capsule or AH were analyzed by high-performance liquid chromatography–mass spectrometry and relative differences between samples were quantified using the tandem mass tag technique. The distribution of XFM on the capsular surface was visualized by SEM and super-resolution light microscopy. Results A small set of proteins was consistently upregulated in capsule samples from patients with XFS and patients with XFG, including microfibril components fibrillin-1, latent transforming growth factor-β–binding protein-2 and latent transforming growth factor-β–binding protein-3. Lysyl oxidase-like 1, a cross-linking enzyme associated with XFS in genetic studies, was an abundant XFM constituent. Ligands of the transforming growth factor-β superfamily were prominent, including LEFTY2, a protein best known for its role in establishing the embryonic body axis. Elevated levels of LEFTY2 were also detected in AH from patients with XFG, a finding confirmed subsequently by ELISA. Conclusions This analysis verified the presence of suspected XFM proteins and identified novel components. Quantitative comparisons between patient samples revealed a consistent XFM proteome characterized by strong expression of fibrillin-1, lysyl oxidase-like-1, and LEFTY2. Elevated levels of LEFTY2 in the AH of patients with XFG may serve as a biomarker for the disease.

The uploaded files aresupplementary tables for a submitted manuscript in NAR Genomics and Bioinformatics journal Manuscript ID: NARGAB-2019-089 Manuscript title:MS/MS in silico subtraction-based proteomic profiling as an approach to facilitate disease gene discovery: application to lens development and cataract Authors: Aryal, Sandeep; Anand, Deepti; Hernandez, Francisco; Weatherbee, Bailey; Huang, Hongzhan; Reddy, Ashok; Wilmarth, Phillip; David, Larry; Lachke, Salil* *Corresponding author:Salil A. Lachke, Department of Biological Sciences, University of Delaware, 105 The Green, Delaware Avenue, 236 Wolf Hall, Newark, DE, USA, Tel.: (302) 831 3040; Fax: (302) 831 2281; E-mail: salil@udel.edu Grant support:Supported by National Institutes of Health (NIH) Grant R01 EY021505 to Dr. Salil A. Lachke Keywords:Lens, iSyTE, Proteome, Embryonic lens development, Protein profiling, Database

Figure S1. Prophylactic autophagosome vaccination did not alter CD3 + CD4+ or CD3 + CD4 + FOXP3+ infiltrates from poly-I:C adjuvant only treatment. Figure S2. Overview of custom 4T1 mutation site peptide array. Figure S3. Vaccinated animals displayed increased IgG signals to 15mers and splenocyte IFNγ recognition of 15mer antigens. Figure S4. Increased IgG signal intensity to 4T1 15mers correlated with IgG signals in naïve animals. Figure S5. Antigens from proteins identified by mass spectrometry were not favored in IgG signal intensity increases. Figure S6. Increased post-vaccination IFNγ secretion in response to 4T1 mutation site peptides. Figure S7. Increased post-vaccination IFNγ secretion upon restimulation with 4T1 tumor cells. (DOCX 1638 kb)

Figure S1. Prophylactic autophagosome vaccination did not alter CD3 + CD4+ or CD3 + CD4 + FOXP3+ infiltrates from poly-I:C adjuvant only treatment. Figure S2. Overview of custom 4T1 mutation site peptide array. Figure S3. Vaccinated animals displayed increased IgG signals to 15mers and splenocyte IFNγ recognition of 15mer antigens. Figure S4. Increased IgG signal intensity to 4T1 15mers correlated with IgG signals in naïve animals. Figure S5. Antigens from proteins identified by mass spectrometry were not favored in IgG signal intensity increases. Figure S6. Increased post-vaccination IFNγ secretion in response to 4T1 mutation site peptides. Figure S7. Increased post-vaccination IFNγ secretion upon restimulation with 4T1 tumor cells. (DOCX 1638 kb)

The behavior of a transverse electromagnetic wave propagating in the direction of a uniform magnetic field in an unbounded fully ionized plasma is examined. Linearized kinetic equations with collision terms of the Krook‐Bhatnager‐Gross type extended by Liboff to include interspecies collisional effects are used in the solution of an inhomogeneous wave equation. The region of ion cyclotron resonance is closely investigated, and the transition of the dispersion relation from the low‐temperature collision‐dominated regime to the high‐temperature regime is observed. It is found that moments of the kinetic equations are adequate in the collision‐dominated regime, but the kinetic equation for the ions must be used at higher temperatures. At these higher temperatures a complete solution of the problem requires numerical work near the source plane. Far from the source, explicit solutions for the fields can be written.

A study is made of the linear spatial response of an electron plasma to a localized one‐dimensional electric field, whose frequency ω is low compared with the electron‐collision frequency ν. For a fully ionized plasma, both electron‐electron and electron‐ion collisions are included in the calculations, the ions being treated as fixed scatterers. It is shown that the neglect of ion dynamics is justified for a suitable choice of parameters. In the hydrodynamic regime, the response function decays exponentially, with decay length equal to a diffusion length a(ων)−½, where a is electron thermal speed, and its amplitude is proportional to ω3/2. In the kinetic regime, the amplitude is proportional to ω, and the decay is not exponential, with characteristic distance being the mean free path a/ν. For a weakly ionized gas, only electron‐neutral collisions are included; in the hydrodynamic region, the dependence of amplitude and decay length on ω is the same as for the fully ionized gas, but the decay is no longer exponential.