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Trabeculectomy is the gold standard procedure performed in glaucoma when topical medication and laser intervention failed. In a trabeculectomy, number of clinical trials have shown the efficacy of ranibizumab in minimizing extracellular matrix accumulation at the filtering site. Ranibizumab (LucentisTM) is a drug that targets vascular endothelial growth factor (VEGF). However, to date the actual mechanisms of this anti-VEGF in trabeculectomy is still not well understood. Hence, in here we aimed to elucidate the effects of ranibizumab on human Tenon’s fibroblast (HTF) isolated from patients undergoing trabeculectomy. In our previous study, we had reported that ranibizumab reduces the level of spermidine metabolite whereby spermidine is an important polyamine for cell proliferation. For this current study, cultured HTFs were divided into untreated, control IgG, ranibizumab only, difluoromethylornithine (DFMO; inhibitor of spermidine) only and ranibizumab with DFMO. All cells were extracted for PCR array (expression of CDKN1A, CDK2, and CDK4) and protein expression of p53, p21, CDK2, and CDK4 by Western Blot. In here, our result demonstrated that cells treated with ranibizumab or DFMO and cells treated with ranibizumab-DFMO have similar effects as both show increased in p53 and p21. Meanwhile, no significant differences in expression of CDKN1A, CDK2 and CDK4 were observed in all groups. In essence, our findings suggest that ranibizumab action is mediated by p21 and p53.

Excessive scarring of the filtering bleb was reported post-surgery and thus lead to failed trabeculectomy. The excessive scarring occurs due to elevated level of TGF-β1 and thus leading to differentiation of tenon’s fibroblast to myofibroblast. Ursodeoxycholic acid (UDCA), the most hydrophilic bile acid, reported as beneficial agent for the treatment of ocular diseases. This study aimed to investigate whether UDCA is able to reduce the differentiation of fibroblast to myofibroblast in TGF-β1-induced human tenons fibroblast (HTF). Material and Methods: Primary HTFs were obtained and divided into untreated group, TGF-β1 treated group (20 ng/ml), UDCA treated group (100 µΜ) and TGF-β1 co-treated with UDCA (combination) group. All cells were subjected for immunostaining (α-SMA and F-actin expression), PCR array (downstream target of TGF-β1: Nf-κβ1, JUN, TIMP1, SMAD3, SMAD2) and western blot (F-actin expression). The statistically significance was set to p < 0.05 and 4 biological replicates were used. Results: The expression of α-SMA and F-actin suggest that UDCA inhibits the fibrous structure of striated actin induces by TGF-β1 as this actin were highly expressed in TGF-β only group. No significance changes in downstream target genes of TGF-β1 except for Nf-ĸβ1. Interestingly, protein expression of ERK 1/2 and gene expression of Nf-ĸβ1 were significantly decreased in combination group (TGF-β1 with UDCA) compared to TGF-β1 only group. Conclusion: UDCA attenuates the effect of TGF-β1 on human Tenon’s fibroblast by downregulating the expression of α-SMA and F-actin expression which could be mediated by ERK1/2 and NF-ĸβ1.

Pseudomonas aeruginosa is the most common causative organism for contact lens-associated corneal ulcer and is commonly treated with fluoroquinolones. With the emergence of resistant strains, it is important to investigate alternative therapies. Despite well-established efficacy of tazocin against systemic Pseudomonas infections, its topical use for the treatment of Pseudomonas keratitis has not been described, hence this study was aimed to find the ocular permeation of Tazocin and its efficacy in treating keratitis in rabbit eyes.We investigated the ocular permeation of topical tazocin after single drop application in normal rabbit eyes by estimating piperacillin and tazobactam concentrations in cornea, aqueous, and vitreous using a validated LC-MS/MS method. Furthermore, we determined the efficacy of repeated dose administration of tazocin against experimentally induced P. aeruginosa keratitis in rabbits in comparison to moxifloxacin. To determine the efficacy, clinical examination, histopathological examination, and estimation of bacterial load and inflammatory cytokines in cornea were done.Significant corneal concentration of piperacillin and tazobactam was detected in normal rabbit corneas after single dose treatment with tazocin. In rabbits with Pseudomonas-induced keratitis, topical tazocin caused significant clinical and histopathological improvement. This improvement was associated with reduction in corneal bacterial load and inflammatory cytokines. Compared to moxifloxacin 0.5%, tazocin treated group showed greater clinical response which was associated with higher interleukin (IL)-1β, lower tumor necrosis factor (TNF)-α, a comparable level of IL-8, greater reduction in corneal bacterial load, and lesser inflammatory cell infiltration.Tazocin showed good ocular penetration and was effective in treatment of Pseudomonas induced keratitis in rabbits.

The anti-fibrotic properties of ranibizumab have been well documented. As an antagonist to vascular endothelial growth factor (VEGF), ranibizumab works by binding and neutralizing all active VEGF-A, thus limiting progressive cell growth and proliferation. Ranibizumab application in ocular diseases has shown remarkable desired effects, however, to date, its antifibrotic mechanism is not well understood. In this study, we identified metabolic changes in ranibizumab-treated human Tenon&rsquo, s fibroblasts (HTFs). Cultured HTFs were treated for 48 h with 0.5 mg/mL of ranibizumab and 0.5 mg/mL control IgG antibody which serves as a negative control. Samples from each group were injected into Agilent 6520 Q-TOF liquid chromatography/mass spectrometer (LC/MS) system to establish the metabolite expression in both ranibizumab treated cells and control group. Data obtained was analyzed using Agilent Mass Hunter Qualitative Analysis software to identify the most regulated metabolite following ranibizumab treatment. At p-value &lt, 0.01 with the cut off value of two-fold change, 31 identified metabolites were found to be significantly upregulated in ranibizumab-treated group, with six of the mostly upregulated having insignificant role in fibroblast cell cycle and wound healing regulations. Meanwhile, 121 identified metabolites that were downregulated, and seven of the mostly downregulated are significantly involved in cell cycle and proliferation. Our findings suggest that ranibizumab abrogates the tissue scarring and wound healing process by regulating the expression of metabolites associated with fibrotic activity. In particular, we found that vitamin Bs are important in maintaining normal folate cycle, nucleotide synthesis, and homocysteine and spermidine metabolism. This study provides an insight into ranibizumab&rsquo, s mechanism of action in HTFs from the perspective of metabolomics.

Article history: Received on: 07/01/2016 Revised on: 07/02/2016 Accepted on: 24/03/2016 Available online: 30/04/2016 This study was a prospective clinical trial to investigate the effects of adding combined tocotrienol-tocopherol mixed fraction (TTMF) and vitamin C (TTMF+C) supplementation on coronary biomarkers in non-statin and statin treated patients with hypercholesterolaemia (HC) with moderate coronary risk. A total of 35 patients were randomised at baseline into one of two groups, (G1) TTMF+C (320mg TTMF plus 500mg vitamin C) alone daily and (G2) TTMF+C (320mg TTMF plus 500mg vitamin C) plus atorvastatin 10 mg daily. The entire supplementation were taken for 12 months. Fasting serum samples were taken at baseline, 2weeks, 3months, 6months and 12months post-randomisation and analysed for inflammatory biomarkers; high sensitivity Creactive protein (hsCRP) and interleukin-6 (IL6). Combination of TTMF and vitamin C supplementation leads to neutral effects on lipid profiles and inflammation; with no added benefit in statin-treated HC patients with moderate coronary risk. This neutral effects may be attributed to the tocopherol composition in TTMF which could possibly attenuate any potential beneficial effects of tocotrienols. Clinical studies using pure tocotrienols in the absence of tocopherols would further confirm this.

Background Steroid-induced ocular hypertension is currently treated in the same way as primary open-angle glaucoma. However, the treatment is often suboptimal and is associated with adverse effects. We evaluated the oculohypotensive effects of topical trans-resveratrol in rats with steroid-induced ocular hypertension and involvement of adenosine receptors (AR) in intraocular pressure (IOP) lowering effect of trans-resveratrol. Methods The oculohypotensive effect of unilateral single-drop application of various concentrations of trans-resveratrol was first studied in oculonormotensive rats. Concentration with maximum effect was similarly studied in rats with steroid-induced ocular hypertension. Involvement of AR was studied by observing the alterations of IOP in response to trans-resveratrol after pretreating animals with AR subtype-specific antagonists. Additionally, we used computational methods, including 3D modelling, 3D structure generation and protein–ligand interaction, to determine the AR–trans-resveratrol interaction. Results All concentrations of trans-resveratrol produced significant IOP reduction in normotensive rat eyes. Maximum mean IOP reduction of 15.1% was achieved with trans-resveratrol 0.2%. In oculohypertensive rats, trans-resveratrol 0.2% produced peak IOP reduction of 25.2%. Pretreatment with A1 antagonist abolished the oculohypotensive effect of trans-resveratrol. Pretreatment with A3 and A2A AR antagonists produced significant IOP reduction in both treated and control eyes, which was further augmented by trans-resveratrol application in treated eyes. Computational studies showed that trans-resveratrol has highest affinity for A2B and A1, followed by A2A and A3 AR. Conclusion Topically applied trans-resveratrol reduces IOP in rats with steroid-induced ocular hypertension. Trans-resveratrol-induced oculohypotension involves its agonistic activity at the A1 AR.

Anti-Vascular Endothelial Growth Factors (Anti-VEGF) agents have received recent interest as potential anti-fibrotic agents for their concurrent use with trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. The effects of this humanized monoclonal antibody on human Tenon's fibroblast (HTF), the key player of post trabeculectomy scar formation, are not fully understood. This study was conducted to understand the effects of ranibizumab on extracellular matrix production by HTF. The effect of ranibizumab on HTF proliferation and cell viability was determined using MTT assay (3-(4,5-dimethylthiazone-2-yl)-2,5-diphenyl tetrazolium). Ranibizumab at concentrations ranging from 0.01 to 0.5 mg/mL were administered for 24, 48 and 72 h in serum and serum free conditions. Supernatants and cell lysates from samples were assessed for collagen type 1 alpha 1 and fibronectin mRNA and protein level using quantitative real time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). After 48-h, ranibizumab at 0.5 mg/mL, significantly induced cell death under serum-free culture conditions (p 0.05). Ranibizumab caused significant reduction of collagen type 1 alpha 1 (COL1A1) mRNA, but not for fibronectin (FN). Meanwhile, COL1A1 and FN protein levels were found upregulated in treated monolayers compared to control monolayers. Ranibizumab at 0.5 mg/mL significantly reduced cell viability in cultured HTF. From this study, we found that single application of ranibizumab is inadequate to induce the anti-fibrotic effects on HTF, suggesting the importance of adjunctive therapy. Further studies are underway to understand mechanism of actions of ranibizumab on HTF.

Purpose. To report a rare case of nasal chondroma presenting as hypertelorism. Case Report. We report a case of a 16-year-old boy with a large calcified mass arising from the posterior nasal cavity presenting as hypertelorism. Surgical excision was done, and the histopathological examination revealed a chondroma. The hypertelorism resolved postoperatively. Conclusions. Nasal chondroma may also present innocuously as hypertelorism as in this case. (Optom Vis Sci 2015;92:S33YS35)

Previous studies have suggested that combination of Tocotrienol-tocopherol mixed fraction (TTMF) vitamin E and vitamin C supplementation act synergistically in vivo to enhance their anti-oxidant properties. However, additional effects beyond that of anti-oxidation of combined vitamin E and vitamin C in statin-treated high coronary risk patients remains unclear. The aim of this study was to investigate pleiotropic effects of TTMF and vitamin C on inflammatory biomarkers and lipid profile in statin-treated high coronary risk patients. This was a pilot, randomized, double-blind, placebo controlled clinical trial. Twenty nine high coronary risk hypercholesterolaemic subjects were randomized into TTMF+C (160mg TTMF, 75% tocotrienol: 25% alpha-tocopherol plus 500mg vitamin C) intervention or placebo groups for 12 months. Blood samples were collected at entry, baseline, 2 weeks, 3, 6 and 12 months post-randomization for analysis of inflammatory biomarkers [(high sensitive C-reactive protein (hsCRP), interleukin-6 (IL-6), tumour necrosis factor alpha (TNFα)] and serum lipid profile [Total cholesterol (TC), low density lipoprotein cholesterol (LDL-c), high density lipoprotein cholesterol (HDL-c) and triglycerides (TG)]. There was no significant difference observed between TTMF+C and placebo groups with regards to percent change of hsCRP, IL-6 and TNF-α concentrations (p>0.05) and TC, LDL-c, TG and HDL-c (p>0.05) at baseline, 2 weeks, 3, 6 and 12 months of intervention. There was also no significant change seen in the mean concentration of all inflammatory biomarkers and serum lipid profile between TTMF+C and placebo groups at all timelines. TTMF and vitamin C supplementation did not significantly improve the inflammatory status and lipid profile. This could in part be contributed to the masking effect of statin’s pleiotropic properties and the possibility of alpha tocopherol in the TTMF mixture attenuating the beneficial effects of pure tocotrienols.

Nicardipine has been shown to have an anti-atherogenic effect in rabbits given a 2% cholesterol diet. Current evidence suggests that lipid peroxidation plays an important role in atherogenesis. This study examines the effect of nicardipine on lipid peroxidation in rabbits given a 2% cholesterol diet, 8 of these rabbits given nicardipine 0.5 mg/kg twice daily intramuscularly for ten weeks while the remaining untreated 6 were controls. After ten weeks, serum malondialdehyde in the control group was significantly higher compared to their baseline levels (P

AIM To report the long-term efficacy and safety of same site revision trabeculectomy with mitomycin application via a posterior approach. METHODS A noncomparative retrospective case series of consecutive revision trabeculectomies performed for inadequate bleb function between March 2003 and March 2007 by a single surgeon. Surgery involved a posterior/fornix incision with opening of the scleral flap posteriorly at the same site as previous surgery and application of 0.2 to 0.4 mg/mL mitomycin. RESULTS Fifty-seven eyes were followed for an average of 33 ± 15 months. Mean baseline intraocular pressure (IOP) reduced from 21.5 ± 6.5 to 11.2 ± 4.4 and 8 ± 3.6 mm Hg at 1 and 5 years, respectively (P

To determine if an increased cup-to-disc ratio (CDR) and retinal nerve fiber layer (RNFL) loss occur after acute primary angle closure (APAC).Prospective, observational case series.Twenty participants with unilateral APAC provided 20 affected eyes and 20 fellow eyes (controls) for analysis.After initial presentation, participants attended 3 further assessments over a 12-month period (visit 2, within 2 weeks; visit 3, 2-3 months; and visit 4, 6-12 months), in which they underwent the following investigations: Heidelberg Retinal Tomography (Heidelberg Engineering, Dossenheim, Germany), optical coherence tomography of the RNFL and macula, and automated perimetry.Cup-to-disc ratio, optic cup area, neuroretinal rim area, RNFL thickness, macular thickness, and volume.There was no change from visits 2 to 4 in CDR (0.46 ± 0.17 vs. 0.47 ± 0.20; P = 0.94), neuroretinal rim area (1.64 ± 0.55 vs. 1.64 ± 0.57; P = 0.96), or other optic nerve head parameters analyzed in eyes with APAC. The mean overall RNFL thickness decreased from 106.6 ± 17.9 μm to 92.9 ± 18.3 μm between visits 2 and 3 (P0.01) in affected eyes. The superior quadrant RNFL thickness decreased from 134.8 ± 25.9 μm to 113 ± 25.7 μm (P0.01), and the inferior quadrant RNFL thickness decreased from 139.1 ± 28.4 μm to 115.6 ± 24.9 μm (P0.01). There was no significant change in macular thickness or volume.This study demonstrated that an increase in CDR does not occur after APAC that is treated promptly, although RNFL loss does occur.

PURPOSE: To evaluate the effect of the anti-VEGF-A monoclonal antibody bevacizumab on primary human Tenon's capsule fibroblasts (HTFs) in an in vitro model of wound healing. METHODS: Fibroblasts were cultured in RPMI media, and bevacizumab was administered at a concentration ranging from 0.25 to 12.5 mg/mL. Fibroblast viability and cell death were assessed using the MTT colorimetric assay, lactate dehydrogenase assay, BrdU assay, and live/dead assay. Fibroblast contractility was assessed in floating collagen gels. Morphologic changes were assessed by transmission electron microscopy. Antifibrosis activities were compared with 5-fluorouracil. RESULTS: Bevacizumab induced a significant dose-related reduction of HTF cell number at 12.5 mg/mL at 72 hours (P < 0.05). Under serum-free conditions, bevacizumab induced significant fibroblast cell death at concentrations greater than 7.5 mg/mL (P < 0.05). Bevacizumab caused a moderate inhibition of fibroblast gel contraction from baseline (P < 0.05). Scanning electron microscopy revealed marked vacuolization in bevacizumab-treated fibroblasts. CONCLUSIONS: Bevacizumab disrupted fibroblast proliferation, inhibited collagen gel contraction ability, and induced fibroblast cell death at concentrations greater than 7.5 mg/mL in serum-free conditions. These results demonstrated that bevacizumab inhibited a number of fibrosis activities in culture. These activities may underpin the antifibrosis effect proposed in vivo.