bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of retinal ganglion cell gene expression by bHLH transcription factors in the developing and ischemic retinas
Authors: MATTER JM , CHIODINI F , DELAFOSSE L , MATTER‐SADZINSKI L , POURNARAS CJ 
Published: September 2009

Abstract Purpose The loss of retinal ganglion cells (RGC) in the glaucomatous retina exhibits similarities to the pattern of neuronal degeneration detected after experimental ischemia. However, a short episode of retinal ischemia does not provoke damage but rather triggers an endogenous form of neuroprotection. HIFs are bHLH proteins that regulate hypoxic response in ischemic retinas and they are involved in neuroprotection. Hypoxic environments also occur in the developing embryo and create specific niches controlling cell differentiation. Genetic analyses of HIF functions have revealed the importance of oxygen as a key regulator of ontogeny. We have compared the transcriptomes of RGCs in ischemic versus developing retinas. Methods Genome‐wide screens were conducted to identify genes which are expressed in newborn RGCs and growing optic nerve axons and which are up‐ or down‐regulated after venal occlusion by photodynamic thrombosis in the rat retinas. Results Atoh7 is a bHLH protein which is central to the transcriptional network regulating the production of RGCs. Among the targets of Atoh7 there are genes involved in the general metabolism and energy supply – e.g., alpha‐enolase (ENO1), glucose‐6 ‐phosphate isomerase (GPI). These glycolytic enzymes are also targets of HIFs and they are upregulated during hypoxia. To investigate the linkage of glycolysis and mitochondrial activity in RGCs, we monitored by confocal time‐lapse imaging the dynamic distribution of mitochondria in the cell bodies and axons of RGCs that express HIF/Atoh7 targets in developing and ischemic retinas. Conclusion Some gene expression programs involved in differentiating RGCs might be reinitiated in neuroprotection.
Gene therapy mediates cone rescue and rejuvenation in the R91W mutant form of Rpe65‐deficiency mice
Authors: ARSENIJEVIC Y , CRIPPA S , PIGNAT V , SAMARDZIJA M , KOSTIC C 
Published: September 2009

Abstract Purpose Given the advances of gene therapy studies to cure RPE65‐derived Leber Congenital Amaurosis (LCA) (clinical trials phase I), it is of prime importance to examine how cones can be rescued in different mutant contexts. Consequently, we evaluated the effect on retinal activity and cone survival of lentivirus‐mediated gene therapy in the R91W knock‐in mouse model expressing the mutant Rpe65R91W gene. Methods An HIV‐1‐derived lentiviral vector (LV) expressing either the GFP or the mouse Rpe65 cDNA under the control of a 0.8 kb fragment of the human Rpe65 promoter (R0.8) was produced. LV‐R0.8‐RPE65 or GFP was injected into 5‐days‐old (P5) or 1 month‐old R91W mice. Functional and morphological retinal rescues were investigated at 4 months of age. Results Increased light sensitivity was detected by ERG and pupillary light responses in animals injected with LV‐R0.8‐RPE65 at both P5 and 1 month compared to controls. Histological analysis showed improved expression of cone markers and cone outersegment morphology. Furthermore, the density of cones in the region of RPE65 delivery after treatment at P5 reached the wild type level. However, before injection at 1 month of age, only a fraction of the cones (40% of the number found in WT animals) in the Rpe65R91W/R91W mice expressed cone transducin, this fraction increased to 64% after treatment. Moreover, these cones appeared normal. Conclusion We show that lentivirus‐mediated Rpe65 gene transfer is very efficacious in early treatments and still efficient during the course of cone degeneration. Moreover, the treatment at 1 month shows a rejuvenation process of the diseased cones. Thus patient suffering from R91W mutation might benefit from a prolonged therapeutic window.
LOC387715/ARMS2 studies – gene sequencing as a procedure of choice
Authors: WYLEGALA E , TEPER S , NOWINSKA A 
Published: September 2009

Abstract Purpose To search for important mutations in LOC387715/ARMS2 gene. Methods 80 patients with choroidal neovascularization subsequent to age‐related macular degeneration undergoing anti‐VEGF treatment were screened for LOC387715/ARMS2 mutations. PCR followed by gene sequencing was performed. If polymorphism A69S coexisted with R38STOP gene cloning with pGEM‐T vector was done. Results R38STOP mutation was found in 5 patients, A69S in 59 patients, in 3 cases R38STOP coexisted with A69S on the other allele. Due to substantial shortening of the R38STOP translation product, the protein is probably not effective. In patients with A69S polymorphism coexisting with R38STOP there is only A69S effective allele – so patients should be treated as A69S homozygous. Conclusion There is increasing number of studies with A69S variant of ARMS2 but only gene sequencing provide reliable date in these cases. If R38STOP is taken into account, A69S is even more important AMD risk factor. Gene sequencing is advisable in studies with LOC387715/ARMS2.
HSP70 and p62/SQSTM 1 regulate differently autophagy clearance in ARPE‐19 cells
Authors: VIIRI J , HYTTINEN J , RYHäNEN T , SALMINEN A , KAARNIRANTA K 
Published: September 2009

Abstract Purpose Prior to proteolysis, heat shock proteins (HSPs) attempt to refold misfolded proteins. If this process is not successful proteins are degradated in proteasomes or in lysosomes. In the present study, the roles of the HSP70 and the p62/SQSTM 1 in autophagy clearance were evaluated in ARPE‐19 cells after proteasome inhibitor treatment. Methods The HSP70, p62/SQSTM 1 and ubiquitin localization were analyzed by immunofluorescense. Confocal and transmission electron microscopy were used to detect cellular organelles and to analyze the morphological changes. HSP70 and p62/SQSTM 1 levels were modulated using RNA interference techniques. Cell viability was measured by colorimetric assay. Results The proteasome inhibitor MG‐132 evoked the accumulation of perinuclear aggregates positive for HSP70, p62/SQSTM 1 and ubiquitin‐protein conjugates. We observed that the aggregation was reversible: a cessation of proteasome inhibition led to clearance of the deposits via autophagy. Interestingly, p62/SQSTM 1 mRNA depletion delayed autophagy clearance and significantly increased cell death in conjunction with proteasome inhibition. The hsp70 RNA interference did not change autophagy clearance, but increased cell death in response to proteasome inhibition. Conclusion The HSP70 and p62/SQSTM 1 regulate differently autophagy clearance in RPE cells. Autophagy seems to be an important mechanism to clean proteasome inhibitor –induced protein aggregates.
Cost‐effectiveness of the Boston keratoprosthesis
Authors: AMENT JD , STRYJEWSKI TP , DOHLMAN CH 
Published: September 2009

Abstract Purpose Blindness due to corneal disease accounts for approximately 8 million of the 37 million blind in the world. A 2007 study confirmed the global cost‐effectiveness (CE) of cataract surgery and penetrating keratoplasty (PK). We sought to determine the CE of the Boston Keratoprosthesis. Methods In a retrospective chart review, patients who underwent KPro type I surgery in the United States over the last 5 years were identified. Patients had to have a minimum of 2 years of visual acuity follow‐up data. Patients with autoimmune diseases and chemical burns were excluded. Eighty‐three patients were included. CE was determined by cost‐utility analysis (CUA). Complications and additional procedures were incorporated into a weighted cost decision tree. A conservative retention estimate of 85% was assumed at 5 years. Visual acuity values were converted to utilities and determined to be normally distributed per Shapiro‐Wilk testing. Results The mean preoperative utility value was 0.449 increasing to 0.630 postoperatively. The mean incremental utility was 0.182. Amongst our cohort, 65 patients experienced improvement in their BSCVA at 2 years, 13 experienced no improvement, and 5 deteriorated. A total discounted incremental QALY gain of 0.763 was obtained. The total discounted cost associated with this utility equaled $11250 (USD). The cost‐utility of the KPro was $13106 per QALY. The model was assessed using both univariate and multivariate sensitivity analyses. Conclusion The KPro is becoming an established procedure in the US for complex ophthalmic patients, often with numerous comorbidities. The cost effectiveness of KPro was determined in this study to be $13106 USD per QALY. This is comparable to PK, which has been reported in the range of $12000‐16000 USD per QALY.
Macular thickness alterations after cataract surgery determined by optical coherence tomography
Authors: ELEFTHERIADOU M , FRAGISKOU S , KYMIONIS G , PAPADAKI T , PANTELEONTIDIS V , PALLIKARIS IG , TSILIMBARIS M 
Published: September 2009

Abstract Purpose To evaluate macular thickness alterations with optical coherence tomography after phacoemulsification and posterior champer intraocular lens implantation. Methods In this prospective study,201 patients who underwent phacoemulsication(102 men and 99 women)with mean age 65±8years were included.Best corrected visual acuity,complete slit lamp examination with lens and OCT examination were performed in all patients before surgery and at one,three and six months postoperatively.Patients were divided into five groups:Group 1(control group‐100 eyes):patients without any predisponding factors for cystoid macular edema,Group 2(15 eyes):patients with complicated surgery,Group 3(27 eyes):patients with epiretinal membrane,Group 4(35 eyes):patients with diabetes and Group 5(24 eyes):patients with glaucoma. Results The preoperative mean minimal foveal thickness(MMFT)in groups 1 and 2 was 204±24μm and 213±47μm respectively and had no significant changes throughout the follow up period(p>0,05).In groups 3,4 and 5 a significant increase of macular thickness was detected.In group 3 the preoperative MMTF was 248±72μm and at 1,3 and 6 months it was 263±86μm(p=0,01),240±30μm(p=0,18)and 270±64μm(p<0,01)respectively.In group 4 the preoperative MMTF was 219±39μm,after 1month the MMTF increased at 257±78μm(p=0,002),at 3months it was 231±46μm(p=0,005)and at the last examination at 6 months it was 236±49μm(p=0,005).In group 5 the initially MMTF(206±21μm)had significant increase in the first[213±30μm(p=0,07)]and in the third month[223±24μm(p=0,03)]. Conclusion Diabetic retinopathy, epiretinal membranes and glaucoma may predispose to increase in macular thickness after cataract surgery.
Slitlamp integrated OCT, what you can see, is what you can scan
Authors: VERBRAAK FD , STEHOUWER M , DEVRIES H , VANLEEUWEN TG 
Published: September 2009

Abstract Purpose To present results of OCT images captured of the posterior and anterior segment of the eye using the SLSCAN‐1, a new FD‐OCT device integrated into a slit lamp. Methods Patients, seen in the outpatient clinic of the Academic Medical Center, were scanned with the SLSCAN‐1, a newly developed OCT scanning device, integrated into a common slit lamp (figure 1). The OCT is a Fourier Domain OCT system (SLD light source, central wavelength 830 nm, bandwidth 30 nm, 1024 pixel CCD camera, scan speed 5k A‐scans per second, 1024 A‐scans per B‐scan). The posterior segment scans have been captured using a standard indirect ophthalmic lens (Volk). A color fundus photography of the observed area is made at the same time (Topcon camera DC1, resolution = 3.24 Mp). Results For posterior segment imaging, the flexible optical arrangement of the slit lamp and the hand‐held lens (Volk), allows the user to scan large areas of the retina (>45 degrees), depending on the pupil size of the patient. In all patients the macula and optic disk could be visualized without any difficulty. In the anterior segment mode, the SLSCAN‐1 allows imaging of the cornea, anterior chamber, iris and parts of the lens. Conclusion The images made by the SLSCAN‐, new slit lamp integrated FD‐OCT, could be very useful to examine patients directly, both posterior and anterior, during regular slit lamp examinations.
Commercial interest
Optical coherence tomography and biomicroscopical analysis in macular holes
Authors: FRUSCHELLI M , MOTOLESE I , MOTOLESE PA , FREZZOTTI P , MOTOLESE E 
Published: September 2009

Abstract Purpose To assess the usefulness of optical coherence tomography (OCT) for better differential diagnosis of macular holes in comparison with biomicroscopical fundus analysis. Methods We reviewed the files of 25 eyes of 24 patients who were diagnosed as having a macular hole on OCT examination and biomicroscopical fundus analysis. Each eye underwent six radial 3‐mm OCT scans centered on the macula, one 6‐mm vertical and one 6‐mm horizontal scan. Retinal thickness was measured at the foveal center and 750 μm from the center, vertically, and horizontally. The diameter of the macular contour was also measured on vertical and horizontal scans. Results On biomicroscopy only 8 eyes (32%) were diagnosed as having macular hole, while in the remaining 17 eyes (68%) diagnosis of macular hole was possible only performing an OCT examination. Conclusion Optical coherence tomography is very useful in the diagnosis of macular holes compared with classic biomicroscopical analysis.
Does internal longitudinal microstructure of retinal veins change with age in medically healthy persons?
Authors: KOTLIAR KE , KHAROUBI A , SCHMIDT‐TRUCKSÄSS A , HALLE M , LANZL IM 
Published: September 2009

Abstract Purpose We demonstrated previously that roughness of the retinal arterial blood column measured along the vessel axis, termed longitudinal arterial profile (LAP) increases significantly in anamnesticly healthy volunteers with increasing age. Recently we have demonstrated age related changes in LAP of medically healthy persons. Whether longitudinal retinal venous profiles (LVP) are altered with age in this group is investigated. Methods 83 medically healthy volunteers were examined by Dynamic Vessel Analyzer (IMEDOS, Jena, Germany) using stimulation with flickering light. 3 age groups were formed: young (N=28, 30,2±4,3 years), middle age (N=28, 42,3±3,3 years) and seniors (N=27, 64,0±5,0 years). Included in the analysis were volunteers without medical vascular risk factors defined as: blood pressure < 140/90 mmHg, HDL > 35 mg/dl, LDL < 190 mg/dl and glucose levels < 110 mg/dl. Retinal venous diameters were measured along 1 mm vessel segments to obtain LVP. Differences were analyzed using Fourier transformation. Results In all age groups power spectra of LVP do not change during all stages of the venous response to the stimulation. There were no significant differences in LVP between the age groups. Venous diameters in middle‐age group were reduced in comparison to the young group at all stages of vessel reaction (p<0,05). Conclusion Our results indicate that in contrast to retinal arteries retinal veins of validated healthy volunteers do not undergo age related microstructural changes. Hence previously reported age related decrease of retinal venous diameter might have physiologic origin due to reduced blood flow in retinal microcirculation with age, not because of structural changes of venous wall.
Lornoxicam pharmacokinetics in the vitreous humor of albino rabbits
Authors: TSIKA C , TSOURDOU A , TZATZARAKIS M , TSILIMBARIS M , PALLIKARIS IG 
Published: September 2009

Abstract Purpose To assess the elimination half‐life of intravitreal lornoxicam, a non‐steroidal anti‐inflammatory drug (NSAID). Methods Both eyes of 15 rabbits were intravitreally injected with 250 μg of commercially available lornoxicam (for intravenous/intramuscular use, Xefo® 8 IV/IM Injection, Nycomed Hellas S.A.). Six eyes were enucleated at time points 0, 1, 2, 6 and 24 hours after the injection was performed. The eyes were immediately frozen at ‐80°C. The vitreous was eviscerated from the eye and the drug was liquid‐liquid extracted from a 0.4 ml sample. Lornoxicam was isolated by a reversed‐phase High Performance Liquid Chromatography (HPLC) method at retention time 10.7 min and detected at 372 nm. The data were statistically analyzed in order to evaluate the pharmacokinetic parameters of the drug. Results The recovery of lornoxicam after liquid‐liquid extraction was calculated at 69.6% and the limit of determination was 0.1 μg/ml. Statistical analysis revealed that lornoxicam concentrations followed first‐order kinetics with an elimination rate constant of 0.235h‐1 and a half‐life of 3.0 h. Conclusion The determination of the pharmacokinetic characteristics of intravitreal lornoxicam allows the possibility for further investigation of the drug's intraocular behaviour and therapeutic potential.
Subconjunctival injection of bevacizumab (Avastin®) for corneal neovascularization
Authors: DE LA PAZ MF , DE SOUSA F , PONCE C , ALVAREZ J , BARRAQUER RI 
Published: September 2009

Abstract Purpose to study the effects of subconjunctival injection of Bevacizumab on corneal neovascularization. Methods Prospective interventional case series on 7 eyes of 7 patients who underwent subconjunctival injection with Bevacizumab. The following parameters were studied pre‐op, at 1 week, 30, 60 and 90 days post‐op: UCVA, BCVA, pachymetry with OCT, slit lamp examination and photographic imaging. Conjunctival impression cytology pre‐op at 1 week and 9o days was done and complications were also noted. Results Pre‐op diagnoses were: herpetic leucoma (4 eyes), chemical burn (2 eyes), neurotrofic keratopathy (1 eye). An informed off‐label consent form prior to procedure was signed. 1.25 mg of subconjunctival Bevacizumab was injected nearest the area affected. Mean preoperative UCVA and BCVA were 0.86 and 0.44 LogMar units, improved to 0.61 and 0.26 LogMar units at 90 days post‐op, respectively. Central and peripheral pachymetry improved from 532 and 623 microns pre‐op, to 529 and 619 microns at 90 days post‐op, respectively. All slit lamp findings and photographic imaging showed a clear regression of superficial and deep stromal corneal vascularization, with clearing of lipid deposits around the affected areas. No toxic effects were noted on conjunctival impression cytology. Conclusion Subconjunctival injection of Bevacizumab is a safe and effective procedure for the regression of superficial and deep corneal neovascularization. It may be a good alternative for patients prior to performing an optical keratoplasty or for those who are poor candidates for the same.
The neuromuscular junction microenvironment in extraocular and limb muscles
Authors: PEDROSA DOMELLOF F , BRäNNSTRöM T , ANDERSEN PM , MARKLUND SL , LIU J 
Published: September 2009

Abstract Purpose To characterise the components of the neuromuscular junction (NMJ) in normal and pathological extraocular muscles (EOMs) and to assess the dynamics of progressive denervation. Methods Limb and EOM samples from 11 controls,8 ALS patients and from transgenic mice with SOD1 mutations (D90A, G93A) paralleling familiar ALS were processed for immunocytochemistry with antibodies against Schwann cell markers (S‐100, p75, GFAP), gangliosides GD1b and GQ1b/GT1a, neurotrophic factors (BDNF, GDNF, NT‐3, NT‐4) and their receptors, parvalbulmin, nestin, desmin and laminin chains. Results The NMJs of normal EOMs had a different cytoskeleton composition. Differences in the expression of gangliosides GD1b and GQ1b/GT1a, Schwann cell marker S‐100, agrin and nestin at the NMJs were noted in the human ALS EOMs. Parvalbumin was absent or scarce in EOM nerve trunks of ALS patients. The analysis of the time aspects of denervation in the animal models is ongoing. Conclusion The human EOMs in late stages of ALS and the EOMs of the transgenic mice show signs of denervation, although these muscles appear remarkably well preserved. High levels of parvalbumin, proposed to be protective for oculomotor neurons in ALS, are not apparent in advanced stages of the disease. The identification of similar endpoints in the EOMs of patients with D90A mutation and the ALS transgenic mice carrying the same mutation indicates that this is a useful model to study the temporal aspects of progressive denervation in the EOMs, to explore aspects of muscle‐nerve interplay that protect the EOMs in motoneuron disease and to gain further knowledge useful for the development of selective tools to modulate eye muscle function in the treatment of strabismus.
Splicing defect in Calpain12 cDNA of a recessive cataract mutation in rats (cat)
Authors: GRAW J 
Published: September 2009

Abstract Purpose The recessive cataract mutation cat was identified among F2 offspring of Wistar rats having been treated with X irradiation; a corresponding mutant line was established (Léonard & Maisin, Nature 205, 1965, 615‐6). Aim of the study is the identification of the underlying mutation. Methods Homozygous cat rats have been outcrossed to wild‐type Brown Norway rats; heterozygous F1 offspring (without cataracts) have been backcrossed to parental homozygous cat rats. Results Among 461 F2 offspring, we observed in total 223 cataracts (48.5%) at the age of 10 weeks; however, 2/3 of these cataracts were discovered at the age of 3 weeks, and most of the remaining third at the age of 7‐8 weeks. Linkage analysis located the mutation on rat chromosome 1 between the markers D1Got76 (76.2 MB) and D1Rat410 (87.6MB). This large interval contains candidate genes like Six5 and Opa3, which have been excluded by sequence analysis of cDNA from rat lenses. Recently, we identified a major splicing error in lens‐derived cDNA of the gene Capn12 encoding Calpain‐12. Besides exon 1, the transcript contains the complete intron 1; in exon 2 it keeps just a few bases to go to the end of intron 4 and than to exon 5; from there no further deviation from the wild‐type sequence was yet observed. It is predicted that the sequence of the first intron leads to a premature stop codon after 10 new amino acids, which might result in a loss of Calpain‐12 activity. Conclusion Calpains has been proposed for a long time to be involved in cataractogenesis. Here we present for the first time a mutation in a calpain‐encoding gene (Capn12), which is very likely to be responsible for cataract formation.
Genomic (epigenetic) DNA methylation in patients with open‐angle glaucoma
Authors: JÜNEMANN A , LENZ B , REULBACH U , SCHLöTZER‐SCHREHARDT U , REJDAK R , KORNHUBER J , KRUSE F , BLEICH S 
Published: September 2009

Abstract Purpose DNA methylation occurs by transfer of a methyl group to cytosine residues in the CpG1 islands. The extent of DNA methylation positively correlates with the extent of gene inactivation. The present study was performed to investigate whether there is an altered global DNA methylation in patients with open‐angle glaucoma. Methods This prospectice case control study included 59 patients with primary open‐angle glaucoma (POAG, age: 68 (SD 8) years), 54 patients with secondary open‐angle glaucoma due to pseudoexfoliation syndrome (PEXG, age: 72 (SD 8) years), and 53 patients with cataract as controls (age: 69 (SD 11) years). Total DNA was extracted from frozen EDTA‐blood using QIAmp DNA Blood Mini Kit (Qiagen). Global methylation status [DNA methylation in %, 1‐(HpaII/MspI)] was measured with a modified non‐radioactive assay. Statistics: Mann‐Whitney‐Test (2‐tailed), the results are presented as means (SD), significance level p<0.05. Results There was a significantly elevated genomic DNA methylation (in %) 1‐(HpaII/MspI) in peripheral mononuclear cells in patients with POAG (68%, SD 18; U=868, Z = ‐2.79, p=0.005), but not in PEXG (55%, SD 17; U=1049, Z=‐0.57, p=0.57) when compared with healthy controls (55%, SD 24). Thus, the difference between POAG and PEXG was exactly 13% of methylated HpaII/MspI restriction sites. Conclusion Since methylation of DNA is an important epigenetic factor in regulation of gene expression these findings may have implications for a possible subsequent derangement of epigenetic control in patients with POAG. Further studies including gene‐specific analyses are needed to clarify the differences between POAG and PEXG.