Dichloroacetate prevents TGFß-induced epithelial-mesenchymal transition of retinal pigment epithelial cells.

Proliferative retinopathies are associated with formation of fibrous epiretinal membranes. At present, there is no pharmacological intervention for the treatment of retinopathies. Cytokines such as TGFß are elevated in the vitreous humor of the patients with proliferative vitro-retinopathy, diabetic retinopathy and age-related macular degeneration. TGFß isoforms lead to epithelial-mesenchymal transition (EMT) or trans-differentiation of the retinal pigment epithelial (RPE) cells. PI3K/Akt and MAPK/Erk pathways play important roles in the EMT of RPE cells. Therefore, inhibition of EMT by pharmacological agents is an important therapeutic strategy in retinopathy. Dichloroacetate (DCA) is shown to prevent proliferation and EMT of cancer cell lines but its effects are not explored on the prevention of EMT of RPE cells. In the present study, we have investigated the role of DCA in preventing TGFß2 induced EMT of RPE cell line, ARPE-19. A wound-healing assay was utilized to detect the anti-EMT effect of DCA. The expressions of EMT and cell adhesion markers were carried out by immunofluorescence, western blotting, and quantitative real-time PCR. The expression of MAPK/Erk and PI3K/Akt pathway members was carried out using western blotting. We found that TGFß2 exposure leads to an increase in the wound healing response, expression of EMT markers (Fibronectin, Collagen I, N-cadherin, MMP9, S100A4, α-SMA, Snai1, Slug) and a decrease in the expression of cell adhesion/epithelial markers (ZO-1, Connexin 43, E-cadherin). These changes were accompanied by the activation of PI3K/Akt and MAPK/Erk pathways. Simultaneous exposure of DCA along with TGFß2 significantly inhibited wound healing response, expression of EMT markers and cell adhesion/epithelial markers. Furthermore, DCA and TGFß2 effectively attenuated the activation of MAPK/Erk/JNK and PI3K/Akt/GSK3ß pathways. Our results demonstrate that DCA has a strong anti-EMT effect on the ARPE-19 cells and hence can be utilized as a therapeutic agent in the prevention of proliferative retinopathies.

INTRAOCULAR DEXAMETHASONE IMPLANT POSITION IN SITU AND OCULAR HYPERTENSION.

PURPOSE: To determine the association between the dexamethasone implant position in the vitreous cavity and ocular hypertension (OHT). METHODS: Retrospective review of patients with at least one intravitreal dexamethasone implant injection between 2012 and 2016. Patients who had a minimum follow-up for 6 months and documented evidence of the implant position were included in the study. Steroid responders, glaucoma patients, vitrectomized eyes, and eyes with liquefied vitreous were excluded. Relevant data were collected from patient charts. Three positions (P1, P2, and P3) were identified: P1 (in contact with the pars plana/ciliary body region), P2 (anterior to vortex veins), and P3 (posterior to vortex veins). Ocular hypertension was defined as absolute intraocular pressure > 25 mmHg and/or intraocular pressure rise > 10 mmHg. The relationship between implant position and intraocular pressure rise after factoring in other characteristics was the outcome measure. Appropriate statistical analysis was performed. RESULTS: A total of 377 patients (432 eyes; 257 males; 677 injections) were eligible for analysis. The median age was 57.24 (±6.32) years. Eighty-eight eyes had OHT. Of these, 54 eyes had the implant in P1. P1 was associated with high intraocular pressure response (>15 mmHg; P = 0.004) and early (<15 days) onset OHT (r = 0.84, P < 0.001). CONCLUSION: Anterior position of dexamethasone implant in situ increases the risk of OHT.

Comprehensive analysis of maillard protein modifications in human lenses: effect of age and cataract.

In human lens proteins, advanced glycation endproducts (AGEs) originate from the reaction of glycating agents, e.g., vitamin C and glucose. AGEs have been considered to play a significant role in lens aging and cataract formation. Although several AGEs have been detected in the human lens, the contribution of individual glycating agents to their formation remains unclear. A highly sensitive liquid chromatography-tandem mass spectrometry multimethod was developed that allowed us to quantitate 21 protein modifications in normal and cataractous lenses, respectively. N(6)-Carboxymethyl lysine, N(6)-carboxyethyl lysine, N(7)-carboxyethyl arginine, methylglyoxal hydroimidazolone 1, and N(6)-lactoyl lysine were found to be the major Maillard protein modifications among these AGEs. The novel vitamin C specific amide AGEs, N(6)-xylonyl and N(6)-lyxonyl lysine, but also AGEs from glyoxal were detected, albeit in minor quantities. Among the 21 modifications, AGEs from the Amadori product (derived from the reaction of glucose and lysine) and methylglyoxal were dominant.

UVA light-excited kynurenines oxidize ascorbate and modify lens proteins through the formation of advanced glycation end products: implications for human lens aging and cataract formation.

Advanced glycation end products (AGEs) contribute to lens protein pigmentation and cross-linking during aging and cataract formation. In vitro experiments have shown that ascorbate (ASC) oxidation products can form AGEs in proteins. However, the mechanisms of ASC oxidation and AGE formation in the human lens are poorly understood. Kynurenines are tryptophan oxidation products produced from the indoleamine 2,3-dioxygenase (IDO)-mediated kynurenine pathway and are present in the human lens. This study investigated the ability of UVA light-excited kynurenines to photooxidize ASC and to form AGEs in lens proteins. UVA light-excited kynurenines in both free and protein-bound forms rapidly oxidized ASC, and such oxidation occurred even in the absence of oxygen. High levels of GSH inhibited but did not completely block ASC oxidation. Upon UVA irradiation, pigmented proteins from human cataractous lenses also oxidized ASC. When exposed to UVA light (320-400 nm, 100 milliwatts/cm(2), 45 min to 2 h), young human lenses (20-36 years), which contain high levels of free kynurenines, lost a significant portion of their ASC content and accumulated AGEs. A similar formation of AGEs was observed in UVA-irradiated lenses from human IDO/human sodium-dependent vitamin C transporter-2 mice, which contain high levels of kynurenines and ASC. Our data suggest that kynurenine-mediated ASC oxidation followed by AGE formation may be an important mechanism for lens aging and the development of senile cataracts in humans.

Andrographolide reduces proliferation and migration of lens epithelial cells by modulating PI3K/Akt pathway.

Lens epithelial cell proliferation, migration, and transdifferentiation are involved in the development of subcapsular cataracts and postoperative capsular opacification (PCO). PI3K/Akt pathway is involved in the proliferation and migration of lens epithelial cells. Andrographolide is the main bioactive component of Andrographis paniculata and is known to possess anti-proliferative and anti-migratory activities. The purpose of this study is to evaluate the effect of andrographolide on proliferation and migration induced by growth factors (TGF-ß and bFGF) in the lens epithelial cell line, FHL 124. We have also evaluated the role of the PI3K/Akt pathway and its alteration by andrographolide during proliferation and migration of lens epithelial cells. The results showed that andrographolide significantly inhibited proliferation in a dose and time dependent manner. The growth factors, TGF-ß and bFGF, induced migration of lens epithelial cells, which was lowered by andrographolide. The growth factors also up regulated phosphorylated Akt (Ser473) and Akt (Thr308), which was abolished by simultaneous treatment of andrographolide. Similar changes were also observed with the PI3K inhibitor, LY290042. Our findings suggest that andrographolide reduces proliferation, migration, and phosphorylated Akt levels in lens epithelial cells. Hence andrographolide can be utilized for the prevention of PCO.

DOPA and DHN pathway orchestrate melanin synthesis in Aspergillus species.

Melanins are high molecular weight hydrophobic pigments that have been studied for their role in the virulence of fungal pathogens. We investigated the amount and type of melanin in 20 isolates of Aspergillus spp.; A. niger (n = 3), A. flavus (n = 5), A. tamarii (n = 3), A. terreus (n = 3), A. tubingensis (n = 3), A. sydowii (n = 3). Aspergillus spp. were identified by sequencing the internal transcribed spacer (ITS) region. Extraction of melanin from culture filtrate and fungal biomass was done and followed by qualitative and quantitative analysis of melanin pigment. Ultraviolet (UV), Fourier transformed infrared (FT-IR), and electron paramagnetic resonance (EPR) spectra analyses confirmed the presence of melanin. The melanin pathway was studied by analyzing the effects of inhibitors; kojic acid, tropolone, phthalide, and tricyclazole. The results indicate that in A. niger and A. tubingensis melanin was found in both culture filtrate and fungal biomass. For A. tamarii and A. flavus melanin was extracted from biomass only, whereas melanin was found only in culture filtrate for A. terreus. A negligible amount of melanin was found in A. sydowii. The maximum amount of melanin from culture filtrate and fungal biomass was found in A. niger and A. tamarrii, respectively. The DOPA (3,4-dihydroxyphenylalanine) pathway produces melanin in A. niger, A. tamarii and A. flavus, whereas the DHN (1,8-dihydroxynaphthalene) pathway produces melanin in A. tubingensis and A. terreus. It can be concluded that the amount and type of melanin in aspergilli largely differ from species to species.

Real-time dynamic intraocular pressure fluctuations during microcoaxial phacoemulsification using different aspiration flow rates and their impact on early postoperative outcomes: a randomized clinical trial.

PURPOSE: To compare the impact of varying fluidic parameters on intraoperative intraocular pressure (IOP) fluctuations and postoperative outcomes. METHODS: Prospective randomized study of 80 eyes undergoing cataract surgery that were randomly assigned to low (aspiration flow rate: 20 cc/min; bottle height: 90 cm; vacuum: 400 mm Hg) and high (aspiration flow rate: 40 cc/min; bottle height: 110 cm; vacuum: 400 mm Hg) fluidic parameter groups. Real-time dynamic intraoperative IOP was measured during nuclear fragment removal. Mean maximum and minimum IOP and percentage reduction in IOP from maximum were compared between groups. Postoperatively, the rate of change in central corneal thickness and anterior chamber inflammation at days 1 and 7, endothelial cell density at 3 months, and corneal clarity on day 1 were compared. RESULTS: Minimum IOP in the low and high parameters groups was 35 ± 4.0 and 34.5 ± 6.8 mm Hg, respectively. Maximum IOP in the low and high parameters groups was 69 ± 3.0 and 85 ± 1.2 mm Hg, respectively (P < .002). Mean percent reduction from maximum was 59% in the high parameters group compared to 41% in the low parameters group, with the difference being statistically significant (P < .002). Rate of change in central corneal thickness was greater in the high parameters group at postoperative days 1 and 7 (P < .001). Anterior chamber inflammation and corneal clarity on the first postoperative day were significantly better in the low parameters group. CONCLUSIONS: Higher aspiration flow rate and bottle heights are associated with high intraoperative IOPs of up to 85 mm Hg. Prolonged elevated IOP during cataract surgery was found to have more anterior segment inflammation and more edematous corneas.

Intraoperative Performance and Early Postoperative Outcomes Following Phacoemulsification With Three Fluidic Systems: A Randomized Trial.

PURPOSE: To compare intraoperative performance and early postoperative outcomes following phacoemulsification with two systems using active fluidics and one using gravity-based fluidics. METHODS: In this prospective randomized trial, 200 eyes were randomized to the traditional and Active Sentry groups (n = 80 eyes each) where the Centurion Vision System was used with traditional or Active Sentry (Alcon Laboratories, Inc) hand-pieces, respectively, or the Infinit group (n = 40 eyes) where the Infiniti Vision System (Alcon Laboratories, Inc) was used. Within the traditional and Active Sentry groups, there were two subgroups with low (30 mm Hg) or high (55 mm Hg) intraocular pressure (IOP) used. Outcome measures compared were: cumulative dissipated energy (CDE), percentage change in central corneal thickness (CCT) at 1 day, 1 week, and 1 month, anterior chamber cells at 1 day and 1 week, rate of rise and fall of IOP following occlusion break, corneal endothelial cell density (ECD), and macular thickness 6 months postoperatively. RESULTS: CDE was significantly lower in group II compared to the traditional group (2.96 ± 1.4 vs 4.14 ± 2.2, P = .001). With 30 mm Hg IOP, the Active Sentry group had significantly less percentage change in CCT at 1 week postoperatively compared to the traditional handpiece group (0.01% vs 0.02%, P = .008). Incidence of anterior chamber cells less than grade 2 on day 1 was significantly higher in the Active Sentry group (82.9% vs 52%, P = .03). Percentage change in ECD was significantly lower in the Active Sentry group (-0.957 vs -0.98%, P = .005). Significantly faster rise of IOP to baseline following occlusion break was seen in the Active Sentry group. CONCLUSIONS: The use of Active Sentry handpiece was associated with lower CDE, less postoperative increase in CCT, fewer anterior chamber cells, and faster rise of IOP following occlusion break. [J Refract Surg. 2024;40(5):e304-e312.].

Visual outcomes comparing emmetropia vs mini-monovision after bilateral implantation of a nondiffractive extended vision intraocular lens: randomized trial.

PURPOSE: To compare visual outcomes and patient satisfaction after bilateral implantation of a nondiffractive extended vision intraocular lens (IOL) when targeting emmetropia vs mini-monovision. SETTING: Iladevi Cataract & IOL Research Centre, Ahmedabad, India. DESIGN: Prospective, randomized controlled trial. METHODS: Patients undergoing bilateral cataract surgery with an extended vision IOL (Vivity) randomized to group I-IOL implantation with emmetropic target in both eyes or group II-IOL implantation with mini-monovision of -0.5 diopters (D) were included in this study. Outcome measures evaluated 6 months postoperatively were unaided and corrected near visual acuity (UNVA, CNVA) at 40 cm and unaided and corrected distance (UDVA, CDVA) and intermediate (UIVA, CIVA) visual acuity at 66 cm. Mesopic contrast sensitivity, binocular defocus curve, Patient-Reported Spectacle Independence Questionnaire, and satisfaction on the McAlinden questionnaire were also assessed. RESULTS: 70 patients enrolled in this study. 34 and 33 patients in groups I and II, respectively, completed follow-up. Binocular UNVA was significantly better in group II (0.26 ± 0.05 vs 0.22 ± 0.08 logMAR, P = .03). Reading add required in group II was significantly lower. UIVA (0.09 ± 0.06 vs 0.07 ± 0.08 logMAR, P = .15) and UDVA (0.02 ± 0.04 vs 0.02 ± 0.05 logMAR, P = .78) were not significantly different between groups. Mesopic contrast sensitivity was not significantly different between the groups. Binocular defocus curve showed significantly better mean visual acuities between -2.0 D and -3.0 D in group II. Patients in both groups had high levels of spectacle independence, with no patient reporting dysphotopsia. CONCLUSIONS: Binocular UNVA was significantly better, with comparable UDVA and mesopic contrast sensitivity when targeting mini-monovision with the nondiffractive extended vision IOL as compared with targeting binocular emmetropia.

Long-Term Postoperative Outcomes Following Cionni Ring and In-the-Bag Intraocular Lens Implantation in Eyes With Subluxated Lenses.

PURPOSE: Evaluate long-term outcomes following Cionni modified capsule tension ring (MCTR) and in-the-bag intraocular lens (IOL) implantation in subluxated lenses. DESIGN: Retrospective case series. METHODS: Forty-one eyes of 31 patients from Raghudeep Eye Hospital, India, with subluxated lenses who had completed a minimum of 5 years' postoperative follow-up were included. Lens extraction, capsular bag fixation with MCTR using 9-0 polypropylene suture, and in-the-bag IOL implantation were performed in all eyes. Corrected distance visual acuity (CDVA), IOL centration, posterior capsule opacification, glaucoma, and retinal complications were documented at final follow-up. RESULTS: The mean age of the cohort was 20.48 ± 16.46 (SD) years. Twenty-four eyes (58%) were below 15 years of age at the time of surgery. Marfan syndrome accounted for 37% cases. Mean follow-up was 9.89 ± 3.81 (SD) years. Thirty-two eyes (74%) had a CDVA of ≥0.3 logMAR at final follow-up. IOL decentration was noted in 7 eyes (17%), requiring a secondary surgery. The mean duration from primary surgery to resurgery was 8.79 years. Seventeen eyes (41%) required a laser capsulotomy, 88% of which were pediatric eyes. Retinal detachment occurred in 4 eyes (10%), 3 of which had Marfan syndrome. CONCLUSION: Capsular bag fixation with an MCTR using 9-0 polypropylene and in-the-bag IOL implantation had good long-term visual outcomes with an acceptable rate of serious postoperative complications in eyes with subluxated lenses. This approach allows preservation of the natural compartments of the eye and placement of an IOL in its most physiological position. However, considering a 17% rate of IOL decentration requiring surgical intervention, long-term stability with nonbiodegradable suture materials such as polytetrafluoroethylene as well as decentration rates following sutured or sutureless scleral fixation should be compared.

Estrogen mediated protection of cytoskeleton against oxidative stress.

BACKGROUND & OBJECTIVES: Cytoskeletal proteins are deregulated during oxidative stress and cataract formation. However, estrogen which protects against cataract formation and harmful effects of oxidative stress has not been tested on the cytoskeleton of lens epithelial cells (LECs). The current study was undertaken to assess if the protection rendered to LECs by estrogen was mediated by preserving the cytoskeletal proteins. METHODS: Oxidative stress was induced by 50 µM of H 2 O 2 in cultured goat LECs (gLECs) and effect of 1 µM 17ß-estradiol (E 2 ) was tested. After treatment, morphological analysis of cells was carried out using haematoxylin-eosin staining and cell density was also quantified. Cell viability was determined using Hoechst (Ho), YO-Pro (YP) and propidium iodide (PI). F-actin and vimentin were localized using phalloidin and anti-vimentin antibody, respectively, and viewed under fluorescence microscopy. Vimentin was further analysed at protein level by Western blotting. RESULTS: H 2 O 2 led to increased condensation of nucleus, cell death and apoptosis but these were prevented with pre- and co-treatment of E 2 with increase in cell viability (P<0.001). E 2 also prevented H 2 O 2 mediated depolymerization of cytoskeleton but was not able to reverse the changes when given after induction of oxidative stress. INTERPRETATION & CONCLUSIONS: Our findings showed that E 2 helped in preventing deteriorating effect of H 2 O 2 , inhibited cell death, apoptosis and depolymerisation of cytoskeletal proteins in LECs. However, the exact mechanism by which estrogen renders this protection to cytoskeleton of lens epithelial cells remains to be determined.

FLACS vs conventional phacoemulsification by junior cataract surgeons: randomized trial.

PURPOSE: To report intraoperative performance and postoperative outcomes of femtosecond laser-assisted cataract surgery (FLACS) and conventional phacoemulsification (PE) in the hands of junior surgeons. SETTING: Iladevi Cataract & IOL Research Centre, Ahmedabad, Gujarat, India. DESIGN: Prospective single-masked randomized controlled trial. METHODS: 320 eyes (320 patients) undergoing cataract surgery for uneventful cataracts in the hands of junior surgeons were randomized to Group 1-FLACS or Group 2-PE. Intraoperatively, cumulative dissipated energy (CDE) and fluid used were compared. Postoperatively, the following were compared: central corneal thickness (CCT) on day 1, 1 week, and 1 month; corneal clarity (day 1 and 1 week); anterior chamber inflammation (day 1 and 1 week); change in endothelial cell density (ECD) at 6 months postoperatively; and corrected distance visual acuity (CDVA) at 1 week and 1 month postoperatively. RESULTS: 157 and 158 patients in groups 1 and 2 analysed. Intraoperatively, CDE (5.41 ± 2.73 vs 8.83 ± 4.28 in Groups 1 and 2, P = .0001) and fluid used (79.33 ± 33.46 vs 101.82 ± 32.23 mL in Groups 1 and 2, P < .0001) were significantly lesser in Group 1. CCT was significantly higher in Group 2 on day 1 (550.96 ± 33.64 vs 587.70 ± 55.76 µm in Groups 1 and 2, P < .0001) and at 1 week postoperatively (527.94 ± 30.78 vs 545.11 ± 35.17 µm in Groups 1 and 2, P = .001). 72% of eyes had clear corneas on day 1 in Group 1 compared with 39% in Group 2 ( P = .01). Anterior chamber inflammation and CDVA were comparable. Change in ECD was significantly lower (9.3%) in Group 1 vs 12.7% in Group 2, P < .0001. CONCLUSIONS: FLACS showed lower intraoperative CDE, fluid usage, lesser increase in CCT, better early postoperative corneal clarity, and lesser change in ECD at 6 months postoperatively in the hands of junior surgeons during standard cataract surgery.

Employing in silico investigations to determine the cross-kingdom approach for Curcuma longa miRNAs and their human targets.

Background: Plant elements and extracts have been used for centuries to treat a wide range of diseases, from cancer to modern lifestyle ailments like viral infections. These plant-based miRNAs have the capacity to control physiological and pathological conditions in both humans and animals, and they might be helpful in the detection and treatment of a variety of diseases. The present study investigates the miRNA of the well-known spice Curcuma Longa and its prospective targets using a variety of bioinformatics techniques. Results: Using the integrative database of animal, plant, and viral microRNAs known as miRNEST 2.0, nine C. longa miRNAs were predicted. psRNA target service foretells the presence of 23 human target genes linked to a variety of disorders. By interacting with a variety of cellular and metabolic processes, miRNAs 167, 1525, and 756 have been found to be critical regulators of tumour microenvironment. SARS-cov2 and influenza A virus regulation have been connected to ZFP36L1 from miRNA 1525 and ETV5 from miRNA 756, respectively. Conclusions: The current cross-kingdom study offers fresh knowledge about how to increase the effectiveness of plant-based therapies for disease prevention and serves as a platform for in vitro and in vivo research development.

Matrix metalloproteinase-2 and -9 activities in the human lens epithelial cells and serum of steroid induced posterior subcapsular cataracts.

PURPOSE: To evaluate the level of matrix metalloproteinase (MMP)-2 and MMP-9 activities in patients with steroid induced posterior subcapsular cataract (PSC). METHODS: This prospective, observational study comprised of 156 patients having either steroid induced PSC (n=50) or non-steroidal PSC (n=106) were performed to evaluate the level of MMP-2 and MMP-9 activities in the lens epithelial cells (LECs) and the serum. Anterior lens capsules harboring LECs were obtained during phacoemulsification and peripheral blood was collected from patients before administration of anesthesia. Serum was separated by centrifugation at 10,000× g for 15 min at 4 °C. The LECs and serum samples were processed to analyze MMP-2 and MMP-9 activities using succinylated gelatin assay. Quantitative real time-PCR (qRT-PCR) was performed to determine the mRNA levels of MMP-2 and MMP-9 in LECs. The mRNA levels were expressed as a ratio, using the delta-delta method for comparing the relative expression results between cases with steroid induced PSC and cases with non-steroidal PSC. MMP-2 and MMP-9 levels were also compared in the two groups using immunolocalization. RESULTS: The level of MMP-2 and MMP-9 activity was found to be high in LECs and serum of cases with steroid induced PSC. Further in all steroid induced cases, a 1.4 fold increase was observed in MMP-2 activity in LECs and a 1.4 fold increase in MMP-9 activity in the serum. Both qRT-PCR and immunolocalization showed increased expression of MMP-2 and MMP-9 activity. CONCLUSIONS: MMP-2 and MMP-9 activity in both LECs and serum was significantly higher in cases with steroid induced PSC. The possible use of MMP-9 as a non-invasive biomarker in ascertaining the presence of steroid induced PSC should be evaluated using a larger sample size.

Recent trends in miRNA therapeutics and the application of plant miRNA for prevention and treatment of human diseases.

Background: Researchers now have a new avenue to investigate when it comes to miRNA-based therapeutics. miRNAs have the potential to be valuable biomarkers for disease detection. Variations in miRNA levels may be able to predict changes in normal physiological processes. At the epigenetic level, miRNA has been identified as a promising candidate for distinguishing and treating various diseases and defects. Main body: In recent pharmacology, plants miRNA-based drugs have demonstrated a potential role in drug therapeutics. The purpose of this review paper is to discuss miRNA-based therapeutics, the role of miRNA in pharmacoepigenetics modulations, plant miRNA inter-kingdom regulation, and the therapeutic value and application of plant miRNA for cross-kingdom approaches. Target prediction and complementarity with host genes, as well as cross-kingdom gene interactions with plant miRNAs, are also revealed by bioinformatics research. We also show how plant miRNA can be transmitted from one species to another by crossing kingdom boundaries in this review. Despite several unidentified barriers to plant miRNA cross-transfer, plant miRNA-based gene regulation in trans-kingdom gene regulation may soon be valued as a possible approach in plant-based drug therapeutics. Conclusion: This review summarised the biochemical synthesis of miRNAs, pharmacoepigenetics, drug therapeutics and miRNA transkingdom transfer.

Late onset corneal decompensation following retained lens fragment in anterior chamber years after uneventful phacoemulsification.

PURPOSE: To report a case of late onset corneal decompensation following cataract surgery due to retained lens fragment in anterior chamber. OBSERVATIONS: A 65 year old female presented with complaint of gradual dimness of vision in left eye since 4 months. She underwent uneventful phacoemulsification with posterior chamber intraocular lens implantation elsewhere 4 years back. On examination, the CDVA in left eye was 20/200. Slit-lamp examination revealed corneal edema with Descemet's folds. She was diagnosed as pseudophakic bullous keratopathy and was being treated with topical steroids, cycloplegics and hyperosmolar agents for the same. She was also counseled about a lamellar corneal transplant. Posterior segment examination was within normal limits. Since the position of the IOL (sulcus versus bag) was not clearly seen ultrasound biomicroscopy (UBM) and anterior segment optical coherence tomography (AS-OCT) imaging was performed to try and better understand the possible cause for corneal decompensation. To our surprise, on both, UBM and ASOCT, a single, retained lens fragment was noted at 6 0'clock in the anterior chamber. AC wash was performed to remove the retained lens fragment. 3 months post AC wash corneal edema resolved completely with improvement in the BCVA to 20/40. CONCLUSION: AND IMPORTANCE: This case highlights the importance of a thorough clinical evaluation supplemented with imaging modalities in order to make a complete diagnosis and eventually achieve better outcomes for the patient. In any case of unexplained corneal edema, either in the early or late postoperative period, UBM and ASOCT can become very helpful to determine the underlying cause.

Silibinin suppresses TGFß2-induced lens epithelial cell migration and epithelial-mesenchymal transition.

Growth factor-induced migration of lens epithelial cell (LEC) toward the posterior of lens capsule bag and their epithelial-mesenchymal transition (EMT) is the key process involved in the pathogenesis of posterior capsular opacification (PCO). Silibinin, a natural flavonolignan, confers therapeutic effects to different cells by regulation of signalling pathways; however, its role in the prevention of migration and EMT of LECs is yet to be analysed. In this study, the inhibitory capabilities of silibinin on migration and EMT were analysed in response to TGFß2 stimulation in HLE B-3 cells. The anti-migratory effect of silibinin was analysed using wound healing assay. Transcriptional and translational expression of genes related to LEC migration, EMT, and transcription factors related to EMT were studied by quantitative real-time PCR and Western blotting. Immunofluorescence analysis was utilized to study the localization of fibronectin. Silibinin reduced the viability of LECs in a concentration-dependent manner and inhibited the wound healing capacity of LECs induced by TGFß2. Silibinin also suppressed alteration in the EMT-related markers such as cytoskeletal proteins, cell adhesion markers, extracellular matrix molecules, and transcription factors. Analysis of downstream signalling revealed that treatment with silibinin decreased phosphorylated Akt (Ser473, Thr308), PDK1 (Ser241), PTEN (Ser380), c-Raf (Ser259), and GSK3ß (Ser9) in TGFß-stimulated cells. The effect of silibinin treatment on phosphorylated Akt resembled that of the PI3K inhibitor LY294002. Our results suggest that silibinin can suppress LEC migration and EMT, which involves the inactivation of the PI3K-Akt signalling pathway. Silibinin might be a good candidate for PCO prevention; however, functional evaluation of silibinin using in vivo models is a pre-requisite.

Fungal scleral keratitis caused by Phomopsis phoenicicola.

We report a case of scleral keratitis caused by Phomopsis phoenicicola. Pterygium surgery was a predisposing factor, and the patient was treated with natamycin and fluconazole eye drops and oral fluconazole. The fungus was identified by sequencing of the internal transcribed spacer (ITS) region of the fungal ribosomal DNA (rDNA) locus and confirmed on the basis of its typical pycnidia and conidia.

Determination of dideoxyosone precursors of AGEs in human lens proteins.

Dideoxyosones (DDOs) are intermediates in the synthesis of advanced glycation endproducts (AGEs), such as pentosidine and glucosepane. Although the formation of pentosidine and glucosepane in the human lens has been firmly established, the formation of DDOs has not been demonstrated. The aim of this study was to develop a reliable method to detect DDOs in lens proteins. A specific DDO trapping agent, biotinyl-diaminobenzene (3,4-diamino-N-(3-[5-(2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanoyl]aminopropyl)benzamide) (BDAB) was added during in vitro protein glycation or during protein extraction from human lenses. In vitro glycated human lens protein showed strong reaction in monomeric and polymeric crosslinked proteins by Western blot and ELISA. Glycation of BSA in the presence of BDAB resulted in covalent binding of BDAB to the protein and inhibited pentosidine formation. Mass spectrometric analysis of lysozyme glycated in the presence of BDAB showed the presence of quinoxalines at lysine residues at positions K1, K33, K96, and K116. The ELISA results indicated that cataractous lens proteins contain significantly higher levels of DDO than non-cataractous lenses (101.9±67.8 vs. 31.7±19.5AU/mg protein, p<0.0001). This study provides first direct evidence of DDO presence in human tissue proteins and establishes that AGE crosslink synthesis in the human lens occurs via DDO intermediates.