bluuebunny/crossref_metadata_2025_split huggingface dataset:
Study of anti‐proliferative activity of ranibizumab, bevacizumab, pegaptanib and aflibercept on fibroblast‐like cell strain in vitro
Authors: LYTVYNCHUK L , S A 
Published: August 2014

AbstractPurpose To evaluate the anti‐proliferative action of drugs that block vascular endothelial growth factor (anti‐VEGF: ranibizumab, bevacizumab, pegaptanib and aflibercept) upon fibroblast‐like cells, as in vitro model for studying their implication on fibroblast‐containing choroidal neovascular membranes (CNV).Methods The cellular anti‐proliferative effects of the four anti‐VEGFs (cellular survival, mitotic‐ and polykaryocyte index, level of apoptosis) were evaluated over 5 days on the fibroblast‐like cell strain L929. Cellular growth‐kinetics indices (specific growth velocity (µ), population number doubling time (td) and reproduction velocity (n)) were calculated and used to reveal dose‐dependence of their anti‐proliferative activity.Results The anti‐VEGFs could inhibit cellular survival, mitotic activity, and increase cellular heterogeneity of L929 cells in a dose‐dependent manner. The anti‐proliferative activity of the anti‐VEGFs was dose‐dependent, while the apoptosis level was proportional to the dose increase in all four cases. The cellular growth‐kinetics distinguished ranibizumab for having less aggressive anti‐proliferative action with increasing doses, which was due to compensation of cell death by proliferation; the rest of the anti‐VEGFs had anti‐proliferative and apoptotic activity prevail the cellular survival. Different concentrations of aflibercept caused insignificant effect upon the cellular mitotic index compared to controls.Conclusion All four anti‐VEGF drugs exhibited marked anti‐proliferative and apoptotic activity upon fibroblast‐like cells in vitro. Overall, the explored effects might explain the fibroblast‐like response to anti‐VEGF drugs in vivo and be the cause for involution of CNVs.
Energy supply is critical for Müller cells ability to transport glutamate
Authors: KOLKO M 
Published: August 2014

AbstractPurpose To study the effect of energy deprivation in Müller cells ability to maintain their function as well as their ability to protect retinal ganglion cells (RGC).Methods The human Müller cell line, MIO‐M1 and primary mouse Müller cells were used to study changes in glutamate uptake, glutamate release, excitatory amino acid transporter (EAAT) protein expression, ATP levels and glycogen content, when cells were compromised from energy. Moreover, a co‐culture system of primary RGC cultures and primary Müller cells, separated by an insert, was used to evaluate the role of Müller cells in RGC survival.Results EAAT1 and EAAT2 proteins were up‐regulated in energydeprived Müller cells and glutamate uptake was significantly increased in the absence of glucose, as opposed to conditions with sodium depletion or in the presence of the EAAT‐inhibitor tfb‐TBOA, in which glutamate uptake was decreased. The intracellular glycogen content decreased in a time‐dependent manner, whereas the ATP levels were sustained following energy deprivation. Co‐cultures of RGC and Müller cells revealed better survival of glutamate treated RGC in the presence of Müller cells compared to controls. Energy deprivation of Müller cells enhanced their ability to protect RGC.Conclusion The present findings revealed an up‐regulation of EAAT1 and EAAT2 in energy compromised Müller cells as well as an increased ability to remove glutamate from the extracellular space. Co‐cultures of RGC and Müller cells revealed a protective role of Müller cells when RGC were exposed to glutamate. Hence, energy failure may result in an increased ability to protect RGC from glutamate‐induced excitotoxicity, whereas malfunction of glutamate uptake in Müller cells may contribute to RGC death.
Activation of cannabinoid receptor CB2 in ARPE‐19 cells stimulates the release of inflammatory cytokines via activation of the ERK pathway
Authors: HYTTI M , PIIPPO N , KORHONEN E , SALMINEN A , KAARNIRANTA K , KAUPPINEN A 
Published: August 2014

AbstractPurpose Endocannabinoid receptors have been suggested to be a possible therapeutical target in age‐related diseases. Particularly CB2, the receptor primarily responsible for the immune modulatory effects of cannabinoids, might be a valuable target in diseases, such as age‐related macular degeneration, where inflammation is known to be a key player. In this study, we have evaluated the effects of CB2 activation on the viability and inflammatory response of retinal pigment epithelial cells.Methods We cultured ARPE‐19 cells until confluent and treated the cells with the selective CB2 agonist JWH‐133. To assess the effect of CB2 activation on oxidatively stressed cells we treated some cells with the lipid peroxidation end product 4‐Hydroxynonenal (HNE) after the JWH‐133 stimulation. The effects of the treatments on cell viability were assessed and inflammatory cytokine expression and signaling protein activation were measured by ELISA.Results Our results show that 10µM JWH‐133 robustly increased the production of interleukin (IL) 6 and IL‐8 in both HNE‐treated and untreated cells. Investigation of the MAPK signaling pathway showed that JWH‐133 at this concentration increased the phosphorylation of ERK1/2.Conclusion Our results show that JWH‐133, a selective agonist of the cannabinoid receptor CB2, stimulates an inflammatory response in ARPE‐19 cells. The release of inflammatory cytokines seems to be mediated by the increased activity of the MAPKinase ERK1/2. Taken together, our results suggest that CB2’s role as a potential therapeutical target in retinal pigment epithelium should be more carefully analyzed in future studies.
Pathophysiologic mechanisms based treatment for RVO related macular edema
Authors: POURNARAS C 
Published: August 2014

AbstractThe retinal vein occlusion (RVO) related to visual loss, results from changes in the blood‐retina barrier leading to the formation of an extracellular retinal edema; and conconmmitant arteriolar vasoconstruction, leading to tissue hypoxia, intracellular retinal edema and neuronal cell death. Patient’s evaluation requires the definition of the pathogenic role of systemic diseases affecting the retinal vessels wall. In addition, hemodynamic modifications of the retinal blood flow and primary and secondary thrombophilias, may trigger the manifestation of the RVO occlusive event. Current treatment of acute RVO aims to restore venous circulation. In that sense, isovolemic hemodilution leads to an increase of ocular blood flow and regression of tissue hypoxia. Grid macular photocoagulation, reversing the inner retinal tissue hypoxia, improves visual prognosis in eyes with macular edema following branch RVO. The intra‐vitreal treatments using anti‐VEGF and anti‐inflammatory drugs, reverse the veins abnormal permeability, relieve the macular edema, either in branch or central RVO, offering new insights for the management of those pathologies.
BRVO treatments in 2014
Authors: PETROPOULOS IK 
Published: August 2014

AbstractUntil recently, laser photocoagulation was the standard of care for branch retinal vein occlusion (BRVO). The recent development of intravitreal pharmacotherapy has revolutionized the management of BRVO and has expanded our treatment options. Intravitreal anti‐VEGF agents (ranibizumab, bevacizumab, aflibercept) and a sustained‐release dexamethasone implant have been shown effective to treat BRVO‐related macular edema by randomized studies and/or case series, offering favorable anatomical and visual outcomes. Still, long‐term repeated injections may be needed and head‐to‐head comparisons of these treatment modalities have not been performed. As for treatment duration, new data suggest that half of the patients may be cured after 2 years from the onset of the disease, whereas the other half will probably need repeated injections for at least 4 years. A better control of the underlying systemic disease (mostly arterial hypertension) and the identification of the proper time to perform laser photocoagulation could further improve prognosis. Safety and efficacy profiles and updated indications of each treatment modality with representative examples will be discussed.
A standardised ‘off the shelf’ substrate for enhanced tissue engineering
Authors: HOPKINSON A , BRANCH MJ , DUA A 
Published: August 2014

AbstractPurpose Amniotic membrane (AM) is a popular ophthalmic tissue engineering adjunct. We previously developed a highly effective thermolysin‐based denuding technique that preserves basement membrane integrity This technique has been validated clinically using fresh frozen AM (FAM). We now propose a standardised dry‐prepared ‘off the shelf’ AM tissue engineering substrate.Methods Dry preserved thermolysin denuded AM (DAM), and FAM denuded with ethylenediaminetetraacetic acid (EDTA), and dispase ‐based methodologies were prepared. Denuding efficiencies were compared using electron microscopy. The effect of denuding on AM molecular composition was investigated and characterised using proteomics. The propensity of DAM to support stem cells was explored using fluorescent immunohistochemistry for defined markers.Results Electron microscopy demonstrated thermolysin denuding efficiency was comparable in FAM and DAM. Proteomic analyses showed effective removal of epithelial cell proteins in DAM, but not EDTA–based denuding techniques. Whilst similar enzymatic activity to thermolysin, mechanical scraping reduces the efficacy of dispase denuding. Collagens IV, VI, periostin, βig‐h3 and VLA‐6 are targets of thermolysin activity. DAM maintains stem cell characteristics and is most effective in preventing differentiation.Conclusion Conventional EDTA and dispase procedures for preparing AM for tissue engineering are ineffective at removing cells whilst preserving the basement membrane. Combining our novel thermolysin denuding and dry preservation techniques improves the overall quality of AM for tissue‐engineered constructs. Therefore, we propose a fist of its kind, stable and dry‐stored ‘off the shelf’ construct for enhanced ocular surface tissue engineering.
Corneal endothelial cells from old donors: differentiation, senescence, proliferative capacities and optimized culture conditions
Authors: HA THI B , HE Z , FOREST F , THURET JY , PISELLI S , PERRACHE C , PEOC'H M , DUMOLLARD JM , ACQUART S , GAIN P , THURET G 
Published: August 2014

AbstractPurpose The intrinsically low proliferative capacity of human corneal endothelial cells (CECs) and their rapid senescence in in vitro culture constitute major limitations for cell bioengineering destined to treat endothelial dysfunctions. In Europe, the mean donor age of 70 adds a supplementary difficulty to the complex equation. Aim: to clarify whether old donor corneas could be however used for cell expansionMethods Morphology and differentiation of central and peripheral CECs were analyzed by immunostaining of ion transport‐related proteins, progenitor and neuronal markers in corneas retrieved in donor >50 years. The proliferative capacity of CECs from organ‐cultured corneas was quantified by EdU proliferation assay. The differences of central vs peripheral CECs were further characterized in vitro by studying their migration, proliferation and differentiation in serum‐containing medium.Results Central CECs highly expressed specific ion transport‐related proteins, and had a high senescence level. On the contrary, CECs located in the periphery or extreme periphery were less specialized, expressed progenitors and neuronal markers and were more prone to endothelial‐mesenchymal transformation in serum‐containing medium. With a new culture sequence that limited proliferation and maintained differentiation, corneas from donors >70 yo, could successfully be used to reconstitute a monolayer of stable endothelial phenotype with ECD of 2000 cells/mm2 but require pooling of corneas.Conclusion This work could allow to “recycle” the 20% of corneas usually discarded by eye banks because of age‐related ECD decline
Ocular chronic graft versus host disease after allogeneic hematopoietic stem cell transplantation – preliminary data
Authors: JEPPESEN H , LINDEGAARD J , SENGELOEV H , JULIAN HO , PRAUSE J , HEEGAARD S 
Published: August 2014

AbstractPurpose Allogeneic hematopoietic stem cell transplantation(HSCT) is a curative therapy for a number of malignant and non‐malignant hematological diseases. Allogeneic HSCT can be performed in several ways; donor cells can be obtained from bone marrow(BMT), peripheral blood(PBSCT) or cord blood(CBT). Ocular chronic graft versus host disease(cGVHD) is a major contributor to long‐term morbidity after HSCT. The purpose of this study was to report the frequency of ocular cGVHD after allogeneic HSCT.Methods Retrospective examination of 233 charts of patients (adults>15years) who underwent consecutive allogeneic HSCT from January 2000‐june 2011 at Copenhagen University Hospital (Rigshospitalet). All allogeneic HSCT in Denmark are performed at this hospital. All patients were examined ophthalmologically before the transplant, yearly after the transplant and ad hoc if any ophthalmic problems occurred. The diagnosis of ocular cGVHD was made according to the NIH consensus criteria.Results Fifty‐eight(25.3%) fulfilled the diagnostic criteria of ocular cGVHD, 16 out of 78(20.5%) in the BMT group, 41 out of 152(27.0%) in the PBSCT group and 1 out of 3(33.3%) in the CBT group. Male:female ratio was 40:18(2.22) (in the cohort 146:87(1.68)). Median time of onset was 24.3 months after HSCT(range 4.9‐146.1). In the cohort median age was 45.2 years(rage 15.2‐71) at transplantation time and 55.5 years(18.43‐69.0) when ocular cGVHD was diagnosed. Median time of follow‐up was 24.8 months(range 0.4‐147.6).Conclusion Out of 233 patients who received allogeneic HSCT, 59(25.3%) developed ocular cGVHD. The median time of onset was 24.3months after transplantation and median age at onset was 55.5 years.
Filtering blebs functionality after trabeculectomy: a clinical and in vivo confocal microscopy study
Authors: VIEIRA L , SANTOS A , LISBOA M , AMARAL A , CUNHA JP , MADURO V , REINA M 
Published: August 2014

AbstractPurpose To evaluate macroscopic and microscopic morphological features of filtering blebs after trabeculectomy with Mitomycin C (MMC) or 5‐Fluorouracil (5‐FU) and correlate them with function.Methods Retrospective case control study of 28 eyes (21 patients) submitted to trabeculectomy (15 with MMC and 13 with 5‐FU) and 11 eyes (10 patients) treated medically. Ophthalmologic examinations included Goldmann applanation tonometry, slit‐lamp examination and photography (Moorfields bleb grading system classification) and in vivo confocal microscopy (Heidelberg Retina Tomograph II, Rostock Cornea Module). Eyes were classiﬁed into 3 groups: control, functioning blebs (Intraocular pressure (PIO)<21mmHg without therapy) and nonfunctioning blebs (PIO<21mmHg with therapy or ≥21mmHg).Results Biomicroscopically, functioning blebs, in comparison to nonfunctioning blebs, showed a significant increase in the central (p=0,009) and maximal (p=0,017) bleb area and a decrease in central (p=0,021) and maximal (p=0,028) bleb vascularization. When comparing successful with failed blebs, in vivo confocal microscopic showed a significant increased number of epithelial microcysts (p=0,014), a liquid content of epithelial microcysts (p=0,008), a lower density of stromal connective tissue (p=0,039) and a decreased number (p=0,041) and diameter (p=0,045) of the vessels. All morphological findings were similar when comparing the usage of MMC with the usage of 5‐FU (p>0,05).Conclusion This study suggests that both biomicroscopy and in vivo confocal microscopy findings can lead to a better understanding and management of filtering blebs.
Self‐tonometry is useful to detect IOP elevations in DALK patients with apparently normal intraocular pressures
Authors: SMEDOWSKI A , TARNAWSKA D , WYLEGALA E 
Published: August 2014

AbstractPurpose To perform increased intraocular pressure (IOP) screening in non‐glaucomatous patients after deep anterior lamellar keratoplasty (DALK) due to keratoconus with normal IOP measured during control visits.Methods Ten non‐glaucomatous patients, who underwent DALK procedure due to keratoconus were included into study. Patients were measuring IOP using self‐tonometer Icare One (Icare, Finland) continuously for 30 days, 3 times per each day. Additionally patients were checked by ophthalmologist 3‐times during follow‐up time in Outpatient Clinic, where applanation tonometry and central corneal thickness measurements were performed. After 30‐days, self‐measured IOP was evaluated for rises above 21 mmHg as well as rises above mean GAT values measured during control visits.Results Mean IOP measured with GAT and adjusted according to CCT was 15 mmHg and ranged between 11 and 19 mmHg. There were no values above 21 mmHg reported during follow‐up time. In Icare One measurements, all patients showed incidents of IOP values higher than measured with GAT, however mean values ranged between 10‐18 mmHg. Six out of 10 patients revealed to have IOP elevations higher than 21 mmHg between control visits (with maximum values up to 46 mmHg), and this elevated values constituted from 3.3 up to 20.0 % of all self‐measurements.Conclusion Self‐tonometry with Icare One might be useful tool to identify patients who develop undetectable IOP increases between control visits. This can help in prevention of vision loss and transplant procedure failure in DALK patients.
Infectious uveitis
Authors: PLEYER U 
Published: August 2014

AbstractThe differential diagnosis of infectious uveitis is broad and an essential step in any initial work‐up. Underlying organisms include all types of infectious agents. The more common infectious causes of uveitis include viruses, T. gondii, T. pallidum, Mycobacterium tuberculosis that will be covered in this course. Based on clinical features further diagnostic tools will be discussed and critically reviewed. In particular newer evolving techniques in the investigations will be included, e.g. intraocular fluid evaluation for polymerase chain testing for the genome and antibody synthesis against the causative organisms.
Subretinal Mononuclear Phagocyte survival and accumulation in AMD
Authors: SENNLAUB F 
Published: August 2014

AbstractThe subretinal space, located between the RPE and the photoreceptor outer segments, is a zone of immune privilege as a consequence of immunosuppressive RPE signals, possibly to limit or subvert inflammatory damage. Age related macular degeneration (AMD) is associated with a breakdown of the subretinal immunosuppressive environment and infiltration and activation of mononuclear phagocytes (MPs, a family of cells that include microglial cells, monocytes, and macrophages) in both late forms of AMD. The molecular mechanisms that provoke the breakdown of local immuno‐suppression in AMD are unknown. Our recent data indicates that Apolipoprotein E (APOE), a lipoprotein that exists in three isoforms in human, participates in this process. We demonstrate that increased APOE secretion from MPs, observed in MPs carrying the AMD‐associated APOe2 allele, leads to prolonged subretinal MP survival and accumulation. We show that subretinal MP accumulation in APOe2 mice is associated with photoreceptor degeneration and excessive experimentally induced choroidal neovascularization, major hallmarks of late AMD. Our results provide a molecular mechanism of local breakdown of immuno‐suppression rationale for the increased AMD‐risk of APOe2 carriers.
Radiation effects on light scattering and cataract formation in the lenses of interventional radiologist and cardiologists
Authors: WEGENER A , HÖCK A , SCHEIDEMANN‐WESP U , CÁMARA R , HAMMER GP , VOßMERBÄUMER U , LASER‐JUNGA H , SINGER S 
Published: August 2014

AbstractPurpose There is little information about exposure to dosages below 500 mSv. This pilot study aimed at investigating the effects of low dose radiation on lens transparency in occupationally exposed interventional cardiologists and radiologists.Methods Physicians who have performed fluoroscopically guided procedures for at least 5 years were recruited in the areas of Cologne‐Bonn and Rhine‐Main together with their non‐exposed colleagues. A questionnaire has been developed to estimate occupational exposure. Exclusion criteria comprised corticosteroid treatment, radiotherapy, chemotherapy and glaucoma. Changes in lens transparency and cataract formation were determined by slit lamp microscopy including LOCS III grading, slit lamp photography and Scheimpflug/Pentacam® imaging.Results The prevalence of lens opacification mainly found in the nuclear region was 52% (95% CI: 30‐74%) among interventional physicians and 31% (95% CI: 9‐61%) among their non‐exposed colleagues. Densitometric analysis of the cornea demonstrated a mean value 52,8% (SD: 7,4) in exposed participants compared to 45,9 % (SD: 7,2) in non‐exposed colleagues. In the lenses layers 4 (deep cortex) and 5 (supra‐nuclear layer) evidenced higher mean densities (40,5% (SD: 3,9%) vs. 31,6% (SD: 6,9%) in layer 4 and 27,5% (SD: 1,9%) vs. 23,7% (SD: 4,3%) in layer 5).Conclusion The data from the ophthalmological examinations demonstrated generally normal findings. However, classification of the mostly small cataracts and densitometric analysis of cornea and lens evidenced that low dosage exposure to ionizing radiation has an effect on corneal and lenticular transparency, albeit on a subclinical level.
Strategies to reduce cytokine levels could suppress PCO progression
Authors: ELDRED JA , SPALTON DJ , WORMSTONE IM 
Published: August 2014

AbstractPurpose Posterior capsule opacification (PCO) develops after cataract surgery. Open bag IOLs, in contrast to current IOL devices, separate anterior and posterior capsules (PC) and are proposed to aid wash out of cytokines in the capsular bag that may promote PCO. The current study assessed the correlation between cytokine availability and cell growth on the human capsular bag.Methods To assess cell growth in different volumes (1.5 Vs 6ml) of Serum‐Free (SF) EMEM we employed a human capsular bag model. We further adapted the capsular bag system, such that radial incisions were made in the anterior capsule, which was then folded back and secured to a dish (a fully open bag model). Cell growth was observed by phase‐contrast microscopy and quantified by ImageJ software. Cytokines were detected by Bioplex suspended bead array at culture day 2. Immunocytochemistry was used to detect myofibroblasts (using the marker alpha smooth muscle actin;αSMA) and cell density.Results Increasing media volume significantly reduced the cell coverage rate on the central PC. A significant cytokine reduction was observed (9 of the 27 assessed) in 6ml cultures compared to 1.5ml counterparts; no cytokines were significantly elevated. At end‐point (28 days) capsular bags maintained in 6ml of media exhibited decreased cell density in the peripheral PC and reduced αSMA expression. Fully open capsular bags cultured in 1.5 or 6ml SF EMEM had limited cell progression onto the PC in both volumes. Fully open capsular bags exposed to 5% human serum demonstrated rapid colonisation of the PCConclusion Reduced cytokine availability is associated with impaired cell growth on the capsular bag. It is likely that open bag devices will restrict cytokine levels and improve the management of PCO.