The tear film is a multi-layer fluid that covers the corneal and conjunctival epithelia of the eye and provides lubrication, nutrients, and protection from the outside environment. Tear fluid contains a high concentration of proteins and has thus been recognized as a potential source of biomarkers for ocular disorders due to its proximity to disease sites on the ocular surface and the non-invasive nature of its collection. This is particularly true in the case of dry eye disease, which directly impacts the tear film and its components. Proteomic analysis of tear fluid is challenging mainly due to the wide dynamic range of proteins and the small sample volumes. However, recent advancements in mass spectrometry have revolutionized the field of proteomics enabling unprecedented depth, speed, and accuracy, even with small sample volumes. In this study using the Orbitrap Fusion Tribrid mass spectrometer, we compared four different mass spectrometry workflows for the proteomic analysis of tear fluid collected via Schirmer strips. We were able to establish a method of in-strip protein digestion that identified >3000 proteins in human tear samples from 11 healthy subjects. Our method offers a significant improvement in the number of proteins identified compared to previously reported methods without pooling samples.
We report a 75-year-old woman with a history of multiple myeloma immunoglobulin D (IgD) variant, who presented with an epibulbar plasmacytoma masquerading as a subconjunctival hemorrhage.Magnetic resonance imaging of the brain and orbits with and without contrast was obtained and surgical biopsy of the subconjunctival lesion was performed; histopathology confirmed the diagnosis of plasmacytoma.Subconjunctival biopsy revealed a plasma cell neoplasm infiltrate in the episcleral layer. The subconjunctival biopsy stained positive for CD138 and lambda-immunohistochemistry in the majority of plasma cells. Histologic findings were consistent with involvement by known IgD plasma cell myeloma where previous bone marrow biopsy demonstrated myeloma cells which stained monoclonally for IgD-lambda light chains.Although plasma cell neoplasms seldom present with ocular manifestations, it is crucial to recognize that these tumors may be associated with multiple myeloma. In patients with known multiple myeloma who present with subconjunctival hemorrhage, close follow-up is highly recommended, as this may be the initial presentation of an ocular plasmacytoma. Although a plasmacytoma is a rare subconjunctival lesion, it should not be immediately excluded from the differential diagnosis of such lesions.
Purpose: The purpose of this study was to discover the aqueous humor proteomic changes associated with visual field indices in glaucoma patients. Methods: Aqueous humor samples were analyzed using Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). The visual fields were analyzed with the Humphrey Visual Field analyzer. Statistical analyses were performed to discover the relationship between the aqueous humor proteins and visual field parameters including Pattern Standard Deviation (PSD), Visual Field Index (VFI), Mean Deviation (MD) and Glaucoma Hemifield Test (GHT). Results: In total, 222 proteins were identified in 49 aqueous humor samples. A total of 11, 9, 7, and 6 proteins were significantly correlated with PSD, VFI, MD, and GHT respectively. These proteins include apolipoprotein D, members of complement pathway (C1S, C4A, C4B, C8B, and CD14), and immunoglobulin family (IKHV3-9, IGKV2-28). Conclusion: Several proteins involved in immune responses (immunoglobulins and complement factors) and neurodegeneration (apolipoprotein D) were identified to be associated with abnormal visual field parameters. These findings provide targets for future studies investigating precise molecular mechanisms and new therapies for glaucomatous optic neuropathy.
Purpose: We aimed to identify clinical factors associated with keratoconus (KC) risk in the All of Us database. Methods: This retrospective matched case–control study utilized patient data from the All of Us Research Program. All patients with a KC diagnosis (n = 572) were enrolled in the study and matched with three controls (n = 1716) based on age ± 1 year, race, ethnicity, and sex. The patients' medical histories, including diabetes, sleep apnea, obesity, smoking, ocular surface disease (encompassing dry eye, eczema, and allergic or atopic conjunctivitis), allergic rhinitis, pregnancy, estrogen-containing medications, tetracyclines, and vitamin C supplementation, were collected using electronic health records. Multivariable odds ratios (ORs) between KC and health history were calculated using the R programming language. Results: The study included 2288 participants with an average age of 58.7 years. All included variables showed a significant positive correlation with KC except smoking history, which showed a negative correlation. The most significant correlations were ocular surface disease (OR = 6.04) and obesity (OR = 1.82). Significant positive associations were also identified for tetracyclines and estrogen-containing medications with KC. Smoking was negatively correlated. Conclusions: In addition to previously known risk factors, patients with a history of increased estrogen exposure and tetracycline usage were more likely to have a KC diagnosis whereas those with a smoking history were less likely. Translational Relevance: Understanding the risk factors for KC, including estrogen exposure and tetracycline medications, enhances our ability to identify at-risk patients and implement earlier screening, diagnosis, and interventions.
Abstract Purpose The aim of the study is to assess the state of glaucoma surgical training in United States ophthalmology residency programs, including experience with microinvasive glaucoma surgery (MIGS). Design The design of the study is anonymous, internet-based national survey. Participants Current United States ophthalmology residents of residency programs accredited by the Accreditation Council for Graduate Medical Education (ACGME). Methods An anonymous survey link was emailed to all 120 accredited United States ophthalmology residency programs inviting residents to participate in an assessment of residency glaucoma surgical experience. Survey responses were collected between January 21, 2019 and March 4, 2019 and analyzed using descriptive statistics. Main Outcome Measures The main outcomes of the study are demographic information, practice intentions, and anticipated primary surgical experience with ACGME-required glaucoma procedures and MIGS procedures, as self-reported by U.S. ophthalmology residents. Results Of the estimated 1,479 U.S. ophthalmology residents, 161 residents participated (10.9%). A total of 118 residents (73.2%) reported any degree of anticipated MIGS primary surgical experience during residency, with the iStent being the most familiar technique. The likelihood of any anticipated MIGS experience during residency was not significantly different by geographic region (p = 0.16), however, anticipated volume varied significantly (p = 0.037). Of the 113 respondents who reported an intention to manage glaucoma surgically in their eventual practice, 25 (22.1%) reported no anticipated primary MIGS experience during residency. 73.3% of residents anticipating MIGS experience anticipated 0 to 10 cases, with 42.9% anticipating less than 5 cases as primary surgeon. Conclusion MIGs are not a required component of the glaucoma surgical curriculum for U.S. ophthalmology residents. Although the majority of ophthalmology residents surveyed intend to manage glaucoma surgically in eventual practice, most receive minimal experience with these novel techniques during residency. Surgical training is variable by geographic region.
Abstract The aqueous humor (AH) is a low-viscosity biofluid that continuously circulates from the posterior chamber to the anterior chamber of the eye. Recent advances in high-resolution mass-spectrometry workflows have facilitated the study of proteomic content in small-volume biofluids like AH, highlighting the potential clinical implications of the AH proteome. Nevertheless, in-depth investigations into the role of AH proteins in ocular diseases have encountered challenges due to limited accessibility to these workflows, difficulties in large-scale AH sample collection and the absence of a reference AH proteomic database. In response to these obstacles, and to promote further research on the involvement of AH proteins in ocular physiology and pathology, we have developed the web-based Aqueous Humor Proteomics Database (AHP DB). The current version of AHP DB contains proteomic data from 307 human AH samples, which were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The database offers comprehensive information on 1683 proteins identified in the AH samples. Furthermore, relevant clinical data are provided for each analyzed sample. Researchers also have the option to download these datasets individually for offline use, rendering it a valuable resource for the scientific community. Database URL: https://ahp.augusta.edu/
Abstract Background Evidence suggests that proteins related to lipid metabolism, such as apolipoproteins, play an important role in the maintenance of normal vision. While several members of the apolipoprotein family are abundant in human aqueous humor (AH), their study remains difficult due to the AH’s small volume, low protein concentration, and the invasive nature of sample collection. In this study, we report the use of Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) to discover associations between AH apolipoproteins and race, gender, and ocular structure in patients with and without primary open angle glaucoma (POAG). Methods AH samples were collected from 231 patients undergoing phacoemulsification or glaucoma incisional surgery at the Medical College of Georgia, Augusta University and subsequently analyzed via LC-MS/MS. The number of peptide spectrum matches (PSMs) for each protein was used as a semi-quantitative measure of relative protein levels. Parameters related to ocular structure were determined using Optical Coherence Tomography (OCT) and Heidelberg Retinal Tomography (HRT). These data sets were probed for relationships between apolipoprotein levels and POAG, demographics (gender and race), and ocular structure. Results A total of ten apolipoproteins were detected in the 231 collected AH samples, with six detected in 100% of the samples, one detected in almost 57% of the samples and three detected in less than 10% of the samples. The levels of APOA1, APOC3, and APOD were higher among POAG subjects. Stratification by gender and race revealed demographic-specific variations. The levels of five apolipoproteins (APOA1, APOA2, APOA4, APOC3, and APOD) were higher in female POAG patients, whereas no apolipoprotein levels were altered in male POAG patients. The levels of APOA1, APOA2, APOA4, and APOD were increased in glaucomatous African American patients, whereas APOE and APOH levels were decreased in glaucomatous Caucasian patients. We also found distinct associations between apolipoprotein levels and OCT and HRT parameters in patients with and without POAG. Conclusions The intra-population variation in apolipoprotein levels highlights the heterogeneity of glaucoma as a disease, suggesting the importance of personalized treatments. Gender and race-specific alterations may be associated with higher risks of POAG in females and members of the African American population.
Purpose: To determine whether bromfenac as a single agent is just as safe and effective as a corticosteroid in post-operative cataract surgery. Methods: Retrospective chart review compared over 400 eyes undergoing cataract surgery. Two hundred eyes were analyzed in each group. The first group received topical Pred Forte® 1 % (prednisolone acetate 1 %, Allergan) four times daily for two weeks, then tapered over the following three weeks, and the second group had topical Bromday™ (bromfenac 0.09 %, ISTA) as a single agent once daily, two days before surgery and for four weeks after. Outcomes measured were best corrected visual acuity (BCVA), post-operative pain, inflammation, cystoid macular edema (CME), and raised intraocular pressure (IOP). Results: Both groups had excellent vision improvement, to 20/27.2 (Pred Forte group) and 20/26.6 (Bromday group) average BCVA at one month post-operation. Both groups had excellent pain and inflammatory control at the one-day, two-week, and one-month time points. There were two eyes (1 %) in the steroid group with CME and only one eye (0.5 %) in the Bromday group. Most remarkably, when looking at one week post-operation or later, there were 16 eyes (8 %) in the corticosteroid group with an elevation in IOP greater than 5 mmHg above baseline, and only five eyes (2.5 %) in the Bromday group (p=0.02) had IOP elevation not attributable to another cause. When considering those eyes with a history of glaucoma, eight out of 25 (32 %) in the corticosteroid group had an elevated IOP in the first week or later, whereas there were no eyes out of 17 (0 %) in the Bromday group. Conclusions: Bromfenac is a safe and effective non-steroidal anti-inflammatory drug (NSAID) when used in the post-operative management of cataract surgery. It appears to be as effective as a topical steroid, even when used alone, without the risk of elevated IOP.
