Biochemical Analysis of Aqueous Humor in an Elderly Patient with Multiple Myeloma Presenting First as Bilateral Chronic Uveitis.

PURPOSE: To study the biochemical analysis of aqueous humor in a patient with multiple myeloma presenting first as chronic uveitis. METHODS: Observational case report. RESULTS: A 63-year-old healthy woman presented with blurred vision in both eyes for 9 months. Slit-lamp examination showed bilateral conjunctival congestion, corneal oedema, and anterior uveitis. Fundus exam revealed normal optic disc with fine retinal folds in the macula. Serum protein electrophoretogram showed a monoclonal M protein band in the gamma globulin region. The bone marrow biopsy revealed hypercellular marrow with trilineage haematopoiesis and the bone marrow aspirate showed clonal plasma cells >10%, confirming the diagnosis of multiple myeloma. Aqueous fluid showed a differential band in electrophoretic profile of aqueous humor protein that on mass spectrometry analysis was strongly suggestive of immunoglobulin band. CONCLUSION: The biochemical analysis of aqueous humor is another diagnostic test to monitor M protein in patients with multiple myeloma.

Synthesis and Characterization of a Novel Peptide Targeting Human Tenon Fibroblast Cells To Modulate Fibrosis: An Integrated Empirical Approach.

Fibrosis is the primary factor influencing the prognosis of glaucoma post-trabeculectomy surgery, an eye condition characterized by increased intraocular pressure (IOP). Despite advancements in surgical procedures and aftercare, it continues to be a serious impediment. During the clinical intervention of scarring, fibrosis is managed by using topical application of combined antifibrotic drugs (mitomycin C). But still, scarring remains a key problem due to minimal drug penetration and nonbioavailability. In this study, we synthesized a cell-specific peptide for modulating scarring in human tenon fibroblasts undergoing epithelial-mesenchymal transition (EMT). The peptide was also conjugated with mitomycin C in order to investigate the effect of the drug conjugation on human tenon fibroblasts from the nanofiber composite system and to evaluate the fibrosis process. Peptide VRF2019 was identified using a subtractive proteomics approach, including solubility, cell penetration, and amphipathic properties. The peptide structure was determined using circular dichroism spectroscopy. The peptide and drug was conjugated using N-ethyl-N'-(3-(dimethylamino)propyl) carbodiimide/N-hydroxysuccinimide (EDC-NHS) chemistry, and the conjugation efficiency was evaluated using high-pressure liquid chromatography. The conjugated product and polycaprolactone (PCL) were electrospun to form a composite nanofiber, which was tested for cytotoxicity and drug release on human tenon fibroblast cells. The modeled VRF2019 peptide structure formed an α-helical structure with all residues spanning the allowed regions of the Ramachandran plot. Subsequent molecular dynamics simulations also demonstrated its membrane penetration potential. The peptide uptake was also studied in human tenon fibroblast cells. High-pressure liquid chromatography (HPLC) and mass spectrometry measurements confirmed peptide-drug conjugation and stability. Furthermore, scanning electron microscopy (SEM) investigation revealed the structure and size of the PCL composite nanofiber. We infer from early research that the PCL composite nanofiber matrix can greatly increase drug delivery and bioavailability.

RETRACTED: Young adults with myopia have lower concentrations of neuromodulators-dopamine and melatonin in serum and tear.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the authors, as questions have arisen in the interim regarding the validity of some of the methods and therefore the results. The authors have provided the following statement: "We have addressed a question raised by the reader on the higher levels of dopamine and melatonin in serum and tear levels reported in this study in Myopia along with controls. Serum and tear levels of dopamine and melatonin in Myopia cases were estimated by HPLC (UV detector) is reported by us in a higher order of magnitude (ng/mL) in both specimens compared to most of the reported data (pg/mL) that are reported based on LC/MS/MS or kit based immunoassay(pg/mL). The sample extraction protocol that we followed before estimation of the analytes was based on removing protein by precipitation methods with no further purification by liquid/liquid extraction/use of SPE cartridges as reported in other published reports. This could have possibly extracted related compounds, such as the closely related indoles in the case of melatonin extraction; the other Dopamine related metabolites respectively. The possibility of confounding by other metabolites in our estimation has not been ruled out by any targeted analysis. This we agree is a limitation in the specificity of the protocol we have followed that may have contributed to the nmol range of detection. We have done the HPLC method of analysis and detection (UV) and not LC-MS/MS or immunoassays by kit method whose sensitivity and specificity are reportedly higher based on the range of testing reported in serum. The lower detection limit of our protocol was not in pg/mL but in ng/mL. Parity in the levels reported was discussed in the manuscript attributing to the analytical method adopted and interpretation based on relative changes. Amongst ocular fluids, Aqueous humor by similar HPLC analysis reportedly shows up to 200 ng/ml in literature (Alkozi, H. et al 2017). However, based on our serum data, we feel that further validation studies and method comparisons are required. The method of estimation of the neurotransmitters HPLC (UV detection) seems to have influenced the absolute levels of dopamine and melatonin in the cases and controls studied and therefore casts doubts on the validity of these data. Though further interpretations can be made on relative changes, we decide to withdraw our paper, to work further on the method comparison. We sincerely apologize for the inconvenience to the readers."

The 88-kDa Eales' protein in serum is a complex of haptoglobin, complement C3, and galectin-1 as identified by liquid chromatography coupled mass spectrometry.

PURPOSE: Eales' disease (ED), an enigmatic inflammatory disease, affects peripheral retinal veins and thereby vision in males. This study was aimed at identifying and deciphering the role of a novel 88-kDa protein reported in the serum and vitreous of patients with ED. EXPERIMENTAL DESIGN: The purified 88-kDa protein was identified by UPLC coupled ESI-QTOF-MS. The identified proteins were quantified in the serum from 20 ED patients and controls (age and sex matched), respectively by ELISA. The interaction of these proteins was studied using co-immunoprecipitation, western blot, and MS analyses. N-glycosylation of protein was observed by MS and lectin blot. RESULTS: The 88-kDa protein was identified to be a complex of haptoglobin, complement C3, and galectin-1. ELISA results showed a 1.5-fold increase in levels of haptoglobin (p = 0.008), with level of complement C3 unaltered and 1.2-fold decreased serum galectin-1 levels (p = 0.003) in ED patients compared to controls. Co-immunoprecipitation illustrated the interaction between haptoglobin and complement C3. Reduced sialylation and increased ß-1, 6-N-acetyl-glucosamine branched N-glycans were observed in haptoglobin of ED patients. CONCLUSION: The 88-kDa protein, a complex of haptoglobin, complement C3, and galectin-1, may play a potential role in ED pathogenesis while levels galectin-1 and haptoglobin may serve as potential biomarker of ED.

CTR1 silencing inhibits angiogenesis by limiting copper entry into endothelial cells.

Increased levels of intracellular copper stimulate angiogenesis in human umbilical vein endothelial cells (HUVECs). Copper transporter 1 (CTR1) is a copper importer present in the cell membrane and plays a major role in copper transport. In this study, three siRNAs targeting CTR1 mRNA were designed and screened for gene silencing. HUVECs when exposed to 100 µM copper showed 3 fold increased proliferation, migration by 1.8-fold and tube formation by 1.8-fold. One of the designed CTR1 siRNA (si 1) at 10 nM concentration decreased proliferation by 2.5-fold, migration by 4-fold and tube formation by 2.8-fold. Rabbit corneal packet assay also showed considerable decrease in matrigel induced blood vessel formation by si 1 when compared to untreated control. The designed si 1 when topically applied inhibited angiogenesis. This can be further developed for therapeutic application.