Bleb Appearance Research Articles

Type I Collagen Suspension Induces Neocollagenesis and Myodifferentiation in Fibroblasts In Vitro.

The ability of a collagen-based matrix to support cell proliferation, migration, and infiltration has been reported; however, the direct effect of an aqueous collagen suspension on cell cultures has not been studied yet. In this work, the effects of a high-concentration aqueous suspension of a micronized type I equine collagen (EC-I) have been evaluated on a normal mouse fibroblast cell line. Immunofluorescence analysis showed the ability of EC-I to induce a significant increase of type I and III collagen levels, parallel with overexpression of crucial proteins in collagen biosynthesis, maturation, and secretion, prolyl 4-hydroxylase (P4H) and heat shock protein 47 (HSP47), as demonstrated by western blot experiments. The treatment led, also, to an increase of α-smooth muscle actin (α-SMA) expression, evaluated through western blot analysis, and cytoskeletal reorganization, as assessed by phalloidin staining. Moreover, scanning electron microscopy analysis highlighted the appearance of plasma membrane extensions and blebbing of extracellular vesicles. Altogether, these results strongly suggest that an aqueous collagen type I suspension is able to induce fibroblast myodifferentiation. Moreover, our findings also support in vitro models as a useful tool to evaluate the effects of a collagen suspension and understand the molecular signaling pathways possibly involved in the effects observed following collagen treatment in vivo.

Quantitative analysis of functional filtering bleb size using Mask R-CNN.

BackgroundDeep learning has had a large effect on medical fields, including ophthalmology. The goal of this study was to quantitatively analyze the functional filtering bleb size with Mask R-CNN.MethodsThis observational study employed eighty-three images of post-trabeculectomy functional filtering blebs. The images were divided into training and test groups and scored according to the Indiana Bleb Appearance Grading Scale (IBAGS) system. Then, 70 images from the training group were used to train an automatic detection system based on Mask R-CNN and perform a quantitative analysis of the function bleb size. Thirteen images from the test group were used to evaluate the model. During the training process, left and right image-flipping algorithms were used for data augmentation. Finally, the correlation between the functional filtering bleb area and the intraocular pressure (IOP) was analyzed.ResultsThe 83 functional filtering blebs have similar morphological features. According to IBAGS, the functional filtering blebs have a high incidence of E1/E2, H1/H2, and V0/V1. Our Mask R-CNN-based model using the selected parameters achieves good results on the training group after a 200-epoch training process. All the Intersection over Union (IoU) scores exceeded 93% on the test group. The Spearman correlation coefficient between the area of functional filtering blebs and the IOP value was −0.757 (P<0.05).ConclusionsDeep learning is a powerful tool for quantitatively analyzing the functional filtering bleb size. This technique is suitable for use in monitoring post-trabeculectomy filtering blebs in the future.

Euptox A Induces G0 /GI arrest and apoptosis of hepatocyte via ROS, mitochondrial dysfunction and caspases-dependent pathways in vivo.

As a toxin of Ageratina adenophora (A. adenophora), euptox A (9-oxo-10, 11-dehydroageraphorone) is known to cause hepatotoxicity in animals. In this study, we examined the effects of euptox A on mouse liver cells and its underlying mechanisms for the first time. We found that euptox A induced liver cell cycle arrest and apoptosis in a dose-dependent manner mainly by mitochondria -related pathways, with the affected cells characterized by the appearance of DNA fragmentation, membrane blebbing, and chromatin condensation. The results showed that euptox A similarly induced hepatocyte G0 /GI arrest and apoptosis mainly by ROS accumulation and mitochondria-mediated and caspase-dependent pathways, elucidated by the loss of mitochondrial membrane potential, release of cytochrome C and AIF, activation of caspase-3/-9, Bax, as well as suppression of Bcl-2. This paper will provide new insights into the mechanisms involved in liver toxicity caused by euptox A in mice.

Bleb grading by photographs versus bleb grading by slit-lamp examination.

Using a bleb-grading system clinically facilitates long-term follow-up of patients with previous glaucoma surgery. Clinical evaluation of these patients can be challenging for untrained ophthalmologists. Morphological bleb configuration might influence planning of follow-up visits in glaucoma patients due to different and individual prognosis after trabeculectomy. In this study, we compared the MaBAGS (Mainz Bleb Appearance Grading System), a classification system for filtering blebs with other classification systems (MBGS/Moorfields Bleb Grading System, IBAGS/Indiana Bleb Appearance Grading Scale) in reference to usability and reliability and compare it to grading by bleb photographs. Forty-two eyes of 31 patients after trabeculectomy were included. Three observers, two senior and one junior observer, graded all blebs using MaBAGS, MBGS and IBAGS during slit-lamp examination. Bleb photographs were reviewed at least 4weeks after clinical examination. Statistical analysis was performed to determine agreement between the observers using intraclass correlation coefficients. With MaBAGS, excellent and good levels of agreement were found for vascularity indices, Seidel test and transparency. Parameters for area and height yielded moderate agreement, while indices for conjunctival mobility and microcysts failed to show satisfying levels of agreement. Using MBGS resulted in excellent and good interobserver consistency for parameters regarding subconjunctival blood, Seidel test, and central and peripheral vascularity. Height and nonbleb vascularity reached moderate levels of agreement. Agreement for area parameters was low. With IBAGS, good levels of agreement were found for height and vascularity, and moderate for extent. In all grading systems, consistency was considerably better between the two experienced observers compared to the inexperienced grader. MaBAGS shows good reproducibility. Using such a grading system improves precision of the description of a highly variable clinical finding. The reliability of grading by slit-lamp examination exceeds that of grading on photographs.

Photophysical Properties of Emissive Pyrido[3,2-c]carbazole Derivatives and Apoptosis Induction: Development towards Theranostic Agents in Response to Light Stimulus.

Pyridocarbazole moieties are present in many natural products, such as olivacine and ellipticine, and their derivatives are well-known anticancer agents. To develop functional therapeutic and diagnostic compounds, three emissive pyrido[3,2-c]carbazole derivatives, PC-X, containing secondary or tertiary amine groups, were synthesized from an aminoquinoline derivative using a palladium complex as the catalyst. X-ray diffraction analyses revealed that PC-X showed highly planar structures between the pyridine ring and the carbazole framework, exhibiting high fluorescence intensities along with solvatochromic behavior. Imaging of HeLa cells treated with PC-X showed no specific accumulation into the organelles; however, a comparative examination showed that the accumulation in mitochondria was the highest as compared to nuclei and lysosomes. Cytotoxicity analysis using HeLa cells showed that PC-H, containing a secondary amine group showed the highest cytotoxicity (IC50 ≈20 μm) as compared to another PC-X having a tertiary amine group. Colocalization with MitoTracker, a typical mitochondrial staining dye, showed apoptosis-like behavior with remarkable appearance of blebbing during irradiation with near UV light (403 nm), suggesting that the PC-H may not only behave as a fluorescence probe for the imaging organelles, but also as a therapeutic agent for inducing apoptosis in HeLa cells, thereby functioning as a theranostic agent.

Subconjunctival Bevacizumab versus Mitomycin C Adjunctive to Subscleral Trabeculectomy

Background: trabeculectomy was considered gold standard procedure for glaucoma treatment. Mitomycin C (MMC) and bevacizumab (BVZ) has been used to increase trabeculectomy success rates. Objective: to compare the outcome of subscleral trabeculectomy with subconjunctival injection of BVZ with that of trabeculectomy with MMC. Patients and Methods: a prospective, non-randomized, comparative study design was used. Thirty eyes from thirty patients with uncontrolled glaucoma were enrolled. Fifteen eyes underwent subscleral trabeculectomy with intraoperative use of MMC (0.2mg/ml for 3 minutes) (group I) and 15 eyes underwent subscleral trabeculectomy with subconjunctival bevacizumab injection (1.25mg/0.1ml) (group II). The outcome measures were the best-corrected visual acuity, intraocular pressure (IOP), number of IOP-lowering medications, complications, and bleb morphologic features (based on the Indiana Bleb Appearance Grading Scale). Results: thirty eyes of thirty patients fulfilled inclusion criteria and assigned to one of two study groups. Each study group included fifteen eyes of fifteen patients. Participants were 14 males and 16 females. Mean ages of patients assigned to MMC and BVZ were 54.0±7.33 and5.7±6.9 years respectively. The difference in mean age was statistically insignificant among treatment groups (p=0.740). Mean number of pre-operative anti-glaucoma medications used by MMC and BVZ treatment groups were 2.47±0.52 and 2.47±0.52 with no significant difference among treatment groups (p=1.00). Conclusions: adjunctive subconjunctival bevacizumab with trabeculectomy is effective in controlling the IOP profile and the post-operative IOP reduction showed no statistically significant different in both group at all postoperative visits. Comparable success rates were achieved in the two groups.

Rosiglitazone Treatment Prevents Postoperative Fibrosis in a Rabbit Model of Glaucoma Filtration Surgery.

To evaluate the potential antifibrotic effect of rosiglitazone (RSG), a peroxisome proliferator-activated receptor γ (PPARγ)-selective agonist, on subconjunctival fibrosis in a rabbit model of glaucoma filtration surgery (GFS) in vivo, and to investigate the underlying mechanisms in human Tenon's fibroblasts (HTFs) in vitro. GFS were performed on adult male New Zealand white rabbits with chronic ocular hypertension previously established by injections of 2% methylcellulose into the anterior chamber. Rabbits were treated by RSG, mitomycin C (MMC) or 5-fluorouracil (5-FU) intraoperatively. The morphology of filtering blebs was evaluated by Indiana Bleb Appearance Grading Scale (IBAGS) scoring. Expression of profibrotic genes was determined by quantitative PCR, immunoblot, and/or histochemical analysis. In vitro studies were performed in a transforming growth factor (TGF)-β1-based cell model of fibrosis. Autophagy was evaluated by the formation of autophagosomes and autolysosomes using fluorescent and transmission electron microscopy and by expression of key mediators in the autophagic pathway. RSG treatment ameliorated a rebound intraocular pressure (IOP) elevation, prolonged the survival of filtering blebs, and attenuated subconjunctival fibrosis in rabbits following trabeculectomy. Pretreatment of HTFs with RSG inhibited TGF-β1-induced expression of profibrotic genes encoding specificity protein 1, connective tissue growth factor, and α smooth muscle actin. RSG augmented TGF-β1-induced autophagy in HTFs via a beclin1/VPS34-dependent mechanism. Augmentation of autophagy is associated with inhibition of TGF-β1-induced profibrotic gene expression by RSG. RSG treatment prevents subconjunctival fibrosis after GFS by inhibition of profibrotic gene expression through a mechanism involved in promoting autophagy in local fibroblasts. RSG represents a novel antifibrotic drug with the potential to improve the success rate of GFS.

Effect of 2 Novel Sustained-release Drug Release Systems on Bleb Fibrosis: An In Vivo Trabeculectomy Study in a Rabbit Model.

Rabbit model studies suggested better morphology blebs with equal intraocular pressure (IOP) efficacy as a standard mitomycin C (MMC) trabeculectomy using a novel slow-release drug delivery antifibrotic system delivering small quantities of MMC and 5-fluorouracil (5-FU). To evaluate 2 different concentrations of biodegradable poly(lactic-co-glycolic acid) (PLGA) system with 5-FU and MMC (ElutiGLASS) for their ability to reduce fibrosis and compare the results with standard trabeculectomy with MMC in a rabbit model. New Zealand albino rabbits (19) were divided into 3 groups (A, B, C) and standard trabeculectomy operation was performed in the right eye of each rabbit.Group (A) had trabeculectomy with MMC (0.4 mg/mL) applied using a Weck cell sponge; (B) trabeculectomy with slow-release ElutiGLASS (0.23 mg, 5-FU/0.33 μg MMC released over 23 to 30 d); (C) trabeculectomy with rapid release ElutiGLASS (0.45 mg of 5-FU/0.65 μg MMC, released over 5 to 7 d). The rabbits were followed for 3 months before euthanasia. Bleb morphology, vascularity, and fibrosis were less pronounced in groups B and C when compared with group A at 3 months. Group B appears to have a lower and more diffuse bleb appearance compared with the other 2 groups with honeycomb appearance on both clinical examination and ultrasound biomicroscopy imaging with higher percentage of maintained bleb space (83%), less fibrosis than group A while maintaining the same low inflammation score as the other 2 groups on histology. At 3 months, the PLGA polymer had completely disappeared in all rabbits. There were no statistical differences in the degree of IOP reduction or histologic inflammation, among the 3 groups. We successfully created a sustained-release antifibrotic drug delivery system that delivered known dosage of the drugs at doses that are significantly lower than the current standard, and resulted in less fibrosis while maintaining a healthy bleb and equal reduction of IOP. These results are supportive of the antifibrotic effect of the slow-release drug delivery system used in conjunction with trabeculectomy, thus paving the way for human pilot studies to improve and simplify existing surgical techniques for filtering surgeries in glaucoma.

Evaluation of Functional Filtering Bleb Using Optical Coherence Tomography Angiography.

PurposeThis study investigated whether filtering blebs can be evaluated using optical coherence tomography angiography (OCT-A) and compared vascularity parameters with conventional bleb grading systems.MethodsA total of 92 patients with glaucoma, who underwent mitomycin C–augmented trabeculectomy, were enrolled in this study, and 92 eyes were assessed in total. The participants underwent OCT-A in external mode and anterior segment photography for bleb evaluation. For evaluation of bleb vascularity, a blinded observer carefully drew the bleb area on the original OCT-A image using a semiautomated program that calculated the color and brightness densities of the selected area. A blinded observer also classified the grades of the bleb vessels using the Indiana Bleb Appearance Grading Scale (IBAGS) and Moorfields Bleb Grading System (MBGS). The vascularity parameters using OCT-A were compared with the IBAGS and MBGS results. In addition, the correlation between intraocular pressure (IOP) and the bleb vascularity parameters was assessed.ResultsVessel density measured by OCT-A demonstrated excellent inter- and intraobserver reproducibility. The color and brightness densities were positively correlated with the IBAGS and MBGS vascularity scores. There was no difference in accuracy when predicting IOP risk using vascularity scores from the IBAGS and MBGS or when estimating IOP risk using the color and brightness densities on the net reclassification index.ConclusionsBleb evaluation using OCT-A can evaluate vessel vascularity and showed correlation to the IBAGS and MBGS vascularity grading.Translational RelevanceBleb vascularity measurements using OCT-A could potentially provide objective and quantitative vessel parameters for bleb evaluation following trabeculectomy.

Tear film stability after trabeculectomy and its relationship with bleb morphology

Objective: To investigate the tear film stability after trabeculectomy and its relationship with bleb morphology using Optical Quality Analysis System Ⅱ (OQAS Ⅱ). Methods: A cross-sectional study. Glaucoma patients undergoing trabeculectomy in the Eye Hospital of Wenzhou Medical University from November 2011 to November 2016 were invited by telephone to perform optical quality, bleb photography, and break-up time examinations, and history of surgeries and medications was collected. Bleb morphology was graded according to the Indiana bleb appearance grading scale. The tear film stability was the average objective scatter index value measured using OQAS Ⅱ for 10 seconds after blinking minus the baseline objective scatter index. The higher the tear film stability value, the worse the stability. The difference in the tear film stability between the surgical eyes and non-surgical eyes was compared by the Mann-Whitney U test, and the relationships between the optical quality, bleb height, extent and vascularity were compared by the Kruskal-Wallis H test. Results: Sixty-three patients (76 eyes) were enrolled in the study, including 55 surgical eyes and 21 non-surgical eyes. The mean follow-up time was (39.6±26.2) months.In the surgical and non-surgical eyes, the M(Q(R)) of tear film stability was 0.46 (0.86) and 0.23 (0.41), respectively. The tear film stability in the surgical eyes was reduced compared to the non-surgical eyes (P=0.044). The trabeculectomy group was divided into three subgroups according to the height of the filtering bleb: H0 (17 eyes), H1 (24 eyes) and H2-3 (14 eyes). The M(Q(R)) of tear film stability in the three subgroups was 0.40(0.68), 0.70(1.02) and 0.40(1.24), respectively, with no statistically significant difference detected (P=0.481). According to the bleb extent, the surgical group was divided into two subgroups: E0-1 (36 eyes) and E2-3 (19 eyes). The M(Q(R)) of optical quality in the two subgroups was 0.63 (0.78) and 0.26(1.17), respectively, with no significant difference detected (P=0.261). According to the degree of bleb vascularity, the surgical group was divided into three subgroups: V0 (25 eyes), V1 (14 eyes), and V2-3 (16 eyes). The M(Q(R)) of optical quality in the three subgroups was 0.39 (0.69), 0.55 (1.18) and 0.63 (1.24), respectively, with no significant difference (P=0.401). Conclusion: Although tear film stability decrease after trabeculectomy, the decrease is not associated with the bleb morphology. (Chin J Ophthalmol, 2019, 55:214-219).

Effects of craniopharyngioma cyst fluid on neurons and glial cells cultured from rat brain hypothalamus

Craniopharyngiomas (CPs) are rare, epithelial tumors of the central nervous system (CNS) that could lead to manifestation of multiple post-operative symptoms, ranging from hormonal imbalance to obesity, diabetes, visual, neurological and neurocognitive impairments. CP is more frequent in children, and has been reported in middle aged adults as well. In fact, arterial laceration and/or brain stroke which may occur following the removal of some CPs is mainly due to calcification of that CPs along with strong attachments to the blood vessels. The dense oily fluid content of CPs is reported to cause brain tissue damage, demyelination and axonal loss in the hypothalamus; however, its exact effect on different cell types of CNS is still unexplored. In this study, we have collected CP cyst fluid (CCF) from mostly young patients during surgical removal and exposed it 9–10 days in vitro to the primary cultures derived from rat brain hypothalamus for 48 h. A gradual decline in cell viability was noted with increasing concentration of CCF. Moreover, a distinct degenerative morphological transformation was observed in neurons and glial cells, including appearance of blebbing and overall reduction of the cell volume. Further, enhanced expression of Caspase-3 in neurons and glial cells exposed to CCF by immunofluorescence imaging, supported by Western blot experiment suggest CCF induced apoptosis of hypothalamic cells in culture. In this study, we have demonstrated the deleterious effects of the cyst fluid on various cell types within the tumors originating region of the brain and its surroundings for the first time. Taken together, this finding could be beneficial towards identifying the region specific toxic effects of the cyst fluid and its underlying mechanism.

Sustained and targeted episcleral delivery of celecoxib in a rabbit model of retinal and choroidal neovascularization

BackgroundTo evaluate the efficacy of selective episcleral delivery of celecoxib formulated in a sustained-release episcleral exoplant on a model of retinal and choroidal neovascularization induced in rabbits by subretinal injection of matrigel combined with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF).MethodsNine New Zealand white rabbits were randomly assigned to three groups (episcleral celecoxib exoplant, intravitreal bevacizumab injection and control group). The bFGF was mixed with matrigel at a concentration of 10 ug/0.1 mL, and VEGF was mixed with matrigel at a concentration of 2 ug/0.1 mL. Animals assigned to celecoxib or intravitreal bevacizumab groups were treated within 03 days from matrigel injection. Fluorescein angiography (FA) and electroretinography (ERG) were performed 5 days, 2, 4 and 8 weeks after matrigel injection. Persistence or regression of three clinical features (subretinal hyperfluorescence, retinal vascular tortuosity and retinal fibrotic spots) was independently evaluated in each study group at all follow-up periods. Statistical analysis using Fisher’s exact test was performed to compare the frequency of findings at each time point between treated groups and control.ResultsIn all study eyes, matrigel induced the appearance of subretinal blebs and the development of retinal and subretinal neovascularization characterized by progressive and late hyperfluorescence on FA. Persistence of subretinal hyperfluorescence was higher in non-treated (control) animals compared to celecoxib (p = 0.0238) treated animals. The mean b-wave amplitude ratios of ERG recordings did not reveal statistically significant differences between the study groups. Control animals retained in average 40% (± 7%) of the pre-treatment recorded b-wave amplitude, compared to 53% (± 29%) after bevacizumab and 53% (± 17%) after celecoxib treatment.ConclusionIn this rabbit model of retinal and subretinal neovascularization, episcleral celecoxib delivery was demonstrated to significantly inhibit neovascularization. It was also noticed, although not statistically significant, an apparent effect of episcleral celecoxib on preventing tractional retinal detachment secondary to epiretinal fibrovascular proliferation. The transscleral delivery of celecoxib combined with sustained-release strategy may have impact in the treatment of retinal and choroidal proliferative diseases.

SCOLICIDAL EFFICACY OF SELENIUM NANOPARTICLES AGAINST PROTOSCOLECES OF HYDATID CYST

The hydatid disease known as hydatidosis or echinococcosis is a cyclozoonotic disease widely distributed in the viscera and other organs of humans and animals. The aim of the present study was to evaluate the efficacy of different concentrations of selenium nanoparticles (Se NPs) (100, 250 and 500μg/ml), albendazole sulfoxide (ABZ sulfoxide) (50μg/ml) andNaCl 0.9% as control with different exposure times (10, 20, 30, 60 min and 3 days) on protoscoleces of Echinococcus granulosus (E. granulosus). They were cultured in test tubes containing RPMI 1640 medium. Viability was examined by use of 0.1% eosin solution and scanning electron microscopy (SEM) was performed for different samples. Exposure to Se NPs at 100μg/ml had a scolicidal effect 100% after 60 min and 3 days only while they killed 93.7%, 95.3%,98.7% at 10, 20 & 30 min application respectively. Also, on exposure to a concentration of 250 μg/ml, the scolicidal activity of Se NPs was 100% after 20, 30, 60 min and 3 days while it was 99.7% after 10 min. On exposure to a concentration of 500μg/ml of Se NPs, all protoscoleces (100%) were killed at all times. Furthermore, exposure to albendazole sulfoxide exhibited partial scolicidal efficacy. SEM of different samples showed obvious alterations with Se NPs 500 μg after 3 days of exposure included invaginated scolex, degenerated rostellum and more severe affection of internal tissue with loss of integrity of the tegument and appearance of blebs.

Five Years' Outcomes of Trabeculectomy with Cross-linked Sodium Hyaluronate Gel Implantation for Chinese Glaucoma Patients.

Background:Trabeculectomy is the most efficient surgical treatment. Prevention failure of bleb cicatrix would lead to unsatisfactory postoperative intraocular pressure (IOP) controlling and unsatisfactory success rate. The aim of this study was to evaluate the 5-year outcomes of trabeculectomy with a cross-linked sodium hyaluronate gel implantation for Chinese glaucoma patients.Methods:This is a prospective, case-controlled study. Patients who were to be applied first-time trabeculectomy in the Department of Ophthalmology of Peking University Third Hospital between 2010 and 2012 were included in the study. Totally, 60 eyes were randomly assigned to the trabeculectomy group (TA group) or the trabeculectomy with cross-linked sodium hyaluronate gel implantation group (TH group). Follow-up was finished at 1 week, 1 month, 3 months, 1 year, 3 years, and 5 years after the operation. The statistical index of demographic data, IOP, bleb shape, and any complications or medications or surgical procedures were recorded and assessed by SPSS 19.0 software through independent t-test, one-way analysis of variance (ANOVA) and Pearson's Chi-square test, respectively.Results:The baseline IOP was comparable between the two groups (t= −1.00, P= 0.32) while the postoperative IOP was significantly lower in the TH group at 1, 3 and 5 years' time points (P = 0.00, P= 0.01 and P = 0.01, respectively). According to the Indiana Bleb Appearance Grading Scale, the height and extent of bleb were better in the TH group at all follow-up time points (P < 0.05), however, the comparison of bleb vascularity showed no statistical difference (P > 0.05). TA group had a higher percentage of complications (13% vs. 3%) compared to TH group; however, there was no statistical difference in the comparison of each statistical item (P > 0.05, respectively). The complete success at 5 years was higher in the TH group than that in the TA group (78% vs. 54%, P = 0.03).Conclusion:Our results suggested that implantation of cross-linked sodium hyaluronate gel with trabeculectomy was more efficient and would improve the prognosis of glaucoma patients.

Treatment Outcomes of Mitomycin C-Augmented Trabeculectomy, Sub-Tenon Injection versus Soaked Sponges, after 3 Years of Follow-up: A Randomized Clinical Trial

To report the 3-year outcome of trabeculectomy with mitomycin C (MMC)-soaked sponges versus intra-Tenon injection of MMC in eyes with uncontrolled primary open-angle glaucoma. Randomized clinical trial. Eighty-two consecutive patients with uncontrolled primary open-angle glaucoma. Participants were randomized either to intra-Tenon injection of 0.1 ml of 0.01% MMC (TI group) or 0.02% subconjunctival application of MMC-soaked sponges (TS group). Patients were followed up for 3 years after surgery. The data for 73 eyes were included in the final analysis. The primary outcome measure was the surgical success, defined as intraocular pressure (IOP) more than 5 mmHg and <21 mmHg, and IOP reduction of 20% or more from baseline, no reoperation for glaucoma, and no loss of light perception vision. Secondary outcome measures were IOP, glaucoma medications, best-corrected visual acuity (VA), bleb morphologic features according to the Indiana Bleb Appearance Grading Scale, complications, and endothelial cell count changes. The cumulative probability of success at 3-year follow-up was 72.2% in the TI group and 65.1% in the TS group (P= 0.30). Uncontrolled IOP was the most common reason for failure. The mean preoperative IOP was 22.4±4.6 mmHg with an average of 3.1±1.0 medications. At 3 years, final IOP was 15.3±3.7 mmHg in the TI group and 16.4±3.5 mmHg in the TS group (P= 0.55). Mean glaucoma number of medications was 0.9±1.1 and 1.1±1.1 in the TI and TS groups, respectively (P= 0.54). Blebs tended to be more diffuse (P= 0.032), less vascularized (P= 0.013), and more shallow (P= 0.012) after intra-Tenon injection. Visual outcomes and endothelial cell changes were similar in both groups (P= 0.47 and P= 0.94, respectively). Although the success rate and IOP reduction were comparable with both techniques, bleb morphologic parameters were more favorable after intra-Tenon injection of 0.1 ml of 0.01% MMC.

Evaluation of intraoperative use of mitomycin C in subscleral trabeculectomy surgery

Objective The aim of the study was to evaluate the effectiveness and complications of mitomycin C (MMC) use in subscleral trabeculectomy surgery. Background MMC is an antimetabolite agent used to limit the scarring process, which is the main cause of failure of subscleral trabeculectomy surgery. However, it may increase the incidence of adverse effects. Patients and methods A prospective interventional study involving 35 eyes of 28 patients having glaucoma was conducted. All of them underwent subscleral trabeculectomy with adjuvant intraoperative use of MMC (0.2 mg/ml) for 2 min, and were followed up for at least 9 months recording intraocular pressure (IOP) testing, best corrected visual acuity, Seidel testing, bleb grading according to the Indiana bleb appearance grading scale, and complications if present. Results The mean IOP decreased from 32.03 mmHg preoperatively to 13.83 mmHg at 9 months postoperatively. The mean IOP drop registered was 56.82%. Overall, 29 (82.9%) eyes ended with complete success, three (8.6%) eyes ended with qualified success, and three (8.6%) eyes failed. The incidence of complications was 51.5%, and most of them were transient and self-limited conditions. Further management, including both medications and minor procedures, improved the postoperative outcomes. Conclusion MMC of 0.2 mg/ml used for 2 min intraoperatively is an effective adjunctive treatment in patients with glaucoma; however, with this dose, it is associated with some complications. So it is a double-edged weapon that should be used wisely, only when indicated, with optimum concentration and duration.

Predictive value of early postoperative IOP and bleb morphology in Mitomycin-C augmented trabeculectomy.

Background: To determine the predictive value of postoperative bleb morphological features and intraocular pressure (IOP) on the success rate of trabeculectomy. Methods: In this prospective interventional case series, we analyzed for one year 80 consecutive primary open angle glaucoma patients who underwent mitomycin-augmented trabeculectomy. Bleb morphology was scored using the Indiana bleb appearance grading scale (IBAGS). Success was defined as IOP ≤15 mmHg at 12 months. We applied a multivariable regression analysis and determined the area under the receiver operating characteristic curve (AUC). Results: The mean age of participants was 62±12.3 years in the success and 63.2±16.3 years in the failure group (P= 0.430) with equal gender distribution (P=0.911). IOPs on day 1, 7 and 30 were similar in both (P= 0.193, 0.639, and 0.238, respectively.) The AUC of IOP at day 1, day 7 and 30 for predicting a successful outcome was 0.355, 0.452, and 0.80, respectively. The AUC for bleb morphology parameters of bleb height, extension, and vascularization, on day 14 were 0.368, 0.408, and 0.549, respectively. Values for day 30 were 0.428, 0.563, and 0.654. IOP change from day 1 to day 30 was a good predictor of failure (AUC=0.838, 95% CI: 0.704 to 0.971) with a change of more than 3 mmHg predicting failure with a sensitivity of 82.5% (95% CI: 68 to 91%) and a specificity of 87.5% (95% CI: 53 to 98%). Conclusions: IOP on day 30 had a fair to good accuracy while bleb features failed to predict success except bleb vascularity that had a poor to fair accuracy. An IOP increase more than 3 mmHg during the first 30 days was a good predictor of failure.

Regulation of apical blebbing in the porcine epididymis.

Apical blebbing, a non-classical secretion mechanism, occurs in the mature porcine epididymis as part of its normal function. Proteins secreted by this mechanism contribute to the modification of the sperm plasma membrane during epididymal transit and are thought to contribute to acquisition of fertilizing ability. However, little is known about the regulation of this secretion mechanism in an invivo model. Previous work demonstrated apical blebbing in the epididymis developed pubertally, suggesting androgens, sperm or other luminal factors regulated this process. Hence, the objective was to evaluate the hypothesized regulation of apical blebbing in the epididymis of pubertal boars by androgens and luminal factors. Androgen receptor blockade (flutamide) and surgical interventions (efferent duct ligation, orchidectomy or transection of the caput epididymis) were used to alter signaling, and the subsequent effects on apical blebbing were evaluated histologically. Apical blebbing was not altered by androgen receptor blockade with flutamide, but was significantly reduced 24 h after efferent duct ligation and after orchidectomy, treatments that eliminated luminal flow from the testis (P < 0.05). Like efferent duct ligation, epididymal transection altered luminal flow without removing the androgen source and significantly reduced the appearance of apical blebbing (P < 0.05). In conclusion, apical blebbing in the porcine epididymis appears to be regulated by luminal factors.

Predictive value of early postoperative IOP and bleb morphology in Mitomycin-C augmented trabeculectomy

Background: To determine the predictive value of postoperative bleb morphological features and intraocular pressure (IOP) on the success rate of trabeculectomy. Methods: In this prospective interventional case series, we analyzed for one year 80 consecutive primary open angle glaucoma patients who underwent mitomycin-augmented trabeculectomy. Bleb morphology was scored using the Indiana bleb appearance grading scale (IBAGS). Success was defined as IOP ≤15 mmHg at 12 months. We applied a multivariable regression analysis and determined the area under the receiver operating characteristic curve (AUC). Results: The mean age of participants was 62±12.3 years in the success and 63.2±16.3 years in the failure group (P= 0.430) with equal gender distribution (P=0.911). IOPs on day 1, 7 and 30 were similar in both (P= 0.193, 0.639, and 0.238, respectively.) The AUC of IOP at day 1, day 7 and 30 for predicting a successful outcome was 0.355, 0.452, and 0.80, respectively. The AUC for bleb morphology parameters of bleb height, extension, and vascularization, on day 14 were 0.368, 0.408, and 0.549, respectively. Values for day 30 were 0.428, 0.563, and 0.654. IOP change from day 1 to day 30 was a good predictor of failure (AUC=0.838, 95% CI: 0.704 to 0.971) with a change of more than 3 mmHg predicting failure with a sensitivity of 82.5% (95% CI: 68 to 91%) and a specificity of 87.5% (95% CI: 53 to 98%). Conclusions: IOP on day 30 had a fair to good accuracy while bleb features failed to predict success except bleb vascularity that had a poor to fair accuracy. An IOP increase more than 3 mmHg during the first 30 days was a good predictor of failure.

Anti-Cancer Activity Of Luvunga Scandens Extract Against Squamous Cell Carcinoma

Introduction: Squamous cell carcinoma is reported as one of the most common types of cancer with increasing numbers of occurrence. Luvunga scandens is a plant possessing many bioactivities and general health effects, yet its anti-proliferative effect is under reported and need to be scientifically evaluated. Materials and Methods: MTT assay was used to assess the cytotoxicity of the plant against human squamous carcinoma cells in addition to the safety assessment for human dermal fibroblast cell line (HDF). The morphological changes of L. scandens treated squamous carcinoma cells has been confirmed by SEM, the apoptosis of the plant against squamous carcinoma cells has been tested using caspase 3/7 assay, followed by cell cycle analysis done using a flowcytometer on squamous carcinoma cells treated with the IC50 dose of L. scandens plant. Results: The plant's extract possesses cytotoxic effect against squamous carcinoma cells with IC50 readings; (methanol= 37.5 mg/mL, dichloromethane= 38 mg/mL, hexane= 37.5 mg/mL), and safe on HDF cells. The SEM results demonstrate that L. scandens treated cells showed an overall change in the cell shape, alteration of surface morphology, absence of microvilli and appearance of blebs. Caspase 3/7 assay results show that L. scandens dichloromethane extract produces the highest level of apoptosis against squamous carcinoma cells. For cell cycle analysis, all the L. scandens treated squamous carcinoma cells show high readings in the sub-G1 phase. Conclusion(s): This in vitro study has proved that L. scandens plant exhibit anti-proliferative effects against Squamous carcinoma cells, hence, it can be considered as a new promising potential anti-cancer therapy.