bluuebunny/crossref_metadata_2025_split huggingface dataset:
New HECplus protocol to further improve the accuracy of pan‐endothelial viability measurement
Authors: Al‐Bourgol Samy , Maurin Corantin , He Zhiguo , Perrache Chantal , Acquart Sophie , Ninotta Sandrine , Forest Fabien , Gain Philippe , Thuret Gilles 
Published: January 2021

AbstractPurposeThe HEC (Hoechst‐Ethidium‐Calcein) labeling, which we described in 2011, is a simple destructive test unanimously adopted to determine the pan‐endothelial viable endothelial cell density (vECD) using image analysis (e.g. using CorneaJ, ImageJ dedicated plugin). Sometimes, the endothelial surface folded by stromal edema remains difficult to analyze. We present here the HECplus technique, which, with an optimization of image acquisition and analysis, further increases the accuracy of vECD determination, particularly in endothelial folds.Methods10 pairs of corneas stored in organoculture at 31°C were analyzed. The endothelial side was incubated with 4 µM Calcein‐AM, 1 µM Ethidium and 11 µM Hoescht 33342 for 45 min at RT. Fluorescence was acquired in 3 channels first with a macroscope, in 3D with an image stacking of the entire graft then with a microscope after flat mounting. A 2 mm plastic pellet emitting constant fluorescence at 510 nm was imaged at the same time to normalize the images. Only one sharp image was reconstructed in ImageJ from the stacks of images. The final standardized images were then analyzed by CorneaJ (HEC group) and then manually by delimiting all viable surfaces on a graphic tablet (HECplus group)ResultsThe vECD was significantly higher by 8.2 ± 1.9% in the HECplus group versus HEC (2059 ± 127 versus 1989 ± 156 cells/mm2). Endothelial cells died preferentially in the top of the folds and were better preserved in the bottom of the folds. While HEC wrongly considered some folds as dead, HECplus allowed the error to be corrected. The folding pattern was similar between the 2 corneas of the same pair but differed from one donor to another. The normalization of images for calcein showed overall differences in the level of fluorescence between corneas, suggesting differences in metabolic activity.ConclusionsWe recommend applying the HECplus protocol when a very high degree of accuracy is required, for example when comparing storage media.
The active storage machine (ASM) allows detecting traces of corneal refractive surgery: case report
Authors: Al‐Bourgol Samy , Ninotta Sandrine , Crouzet Emmanuel , Maurin Corantin , Caroline Trone Marie , Garcin Thibaud , Sophie Gauthier Anne , Thuret Gilles , Gain Philippe 
Published: January 2021

AbstractPurposeThe detection and selection of corneas operated on for refractive surgery (LASIK or photorefractive keratectomy (PKR) by excimer) will become a major concern for eye banks in the coming years because this surgery is often forgotten during the interview with the deceased's relatives. Passive storage in organoculture (OC) prevents the detection of the traces of this surgery because of stromal edema making the cornea less transparent and wrinkled. On the other hand, the active storage machine (ASM) restores intraocular pressure, restores the cornea to its original shape, respects transparency and incorporates non‐invasive controls. It is, therefore, likely to facilitate the detection of refractive surgeries. We present here the case of 2 corneas operated on with PKR and stored successively in OC and ASM.MethodsThe 2 corneas of a 49‐year‐old donor operated 20 years earlier by PKR for ‐2 and ‐3 diopters myopia were stored in OC for 14 days and then placed in ASM for 48 hr. Transparency (transparometer, BiiGC), thickness map and OCT topography (Casia 1, Tomey) were performed under the 2 storage conditions. At the end of the observation, histology and electron microscopy were performed.ResultsTraces of PKR remained unnoticed in OC while they were evident in ASM: central epithelial anomaly, central thinning and flattening of central keratometry were found respectively in transparometry, pachymetry map and OCT topography. Histology and ultrastructure confirmed the absence of Bowman's membrane, pathognomonic of PRK.ConclusionsBy placing the cornea under physiological conditions, and in particular by triggering its deswelling thanks to the intraocular pressure and by restoring its natural curvature, the ASM should allow effective detection of refractive surgery traces.
Visual acuity and contrast sensitivity are slightly modified in young subjects at high genetic risk of developing Alzheimer’s disease
Authors: López‐Cuenca Inés , Elvira‐Hurtado Lorena , Salobrar‐Garcia Elena , Fernández‐Albarral José A. , Lozano María Pilar Rojas , Corral Juan José Salazar , Ramirez Ana Isabel , Ramirez Jose Manuel , De Hoz Rosa 
Published: January 2021

PurposeAlzheimer’s diseases (AD) is the most common cause of dementia characterized by a progressive decline in cognitive function, especially in the memory domain. It is known that its signs can appear decades before to clinical symptoms. In addition to age, genetic studies have identified an association between the presence of the Apoliprotein (ApoE) ɛ4 allele and family history of disease with the late‐onset AD. The aim of the present study was analyzed the possible changes that may occur in visual acuity (VA) and contrast sensitive test (CST) in young subjects with two marked genetic factors risk of developing Alzheimer’s (AD): a first‐degree family history of the disease and being carrier of at least one e4 allele for the ApoE gene.MethodsA case‐control study was conducted in 28 subjects with family history of AD (FH+), ApoE ɛ4 carriers and with a mean age of 53.39 years and 16 age‐matched control subjects without a family history of AD (FH‐), ApoE ɛ4 non‐carriers and with a mean age of 53.94 years. A complete eye examination was performed in all participants, who were free of ocular pathology.ResultsWe found statistical significant increase (p < 0.05) in FH+ ApoE ɛ4 carriers compared to FH‐ ApoE ɛ4 non‐carriers in: I) VA (1.04 ± 0.10, FH+ vs 0.98 ± 0.04, FH‐) and II) CS in the spatial frequency (SF) of 12 cycles/degree (1.68 ± 0.18, FH+ vs 1.54 ± 0.17, FH‐).ConclusionsIn FH+ ApoE ɛ4 carriers, the VA and CST registered changes compared to FH‐ ApoE ɛ4 non‐carriers. In young subjects at high genetic risk of developing AD, an increase of VA and CS in the 12 cicles/degree SF could be the first functional changes to appear.
Innovative ex vivo model of human herpetic keratitis to evaluate antiviral drugs: proof of concept with 0.15% ganciclovir ophthalmic gel
Authors: Courrier Emilie , Maurin Corantin , Bourlet Thomas , Herbepin Pascal , Paul Verhoeven , Gain Philippe , Thuret Gilles 
Published: January 2021

AbstractPurposeWe have recently developed and characterized a unique model of herpetic keratitis reproducing dendritic or geographic ulceration on human corneas stored in our corneal Bioreactor (BR). The BR is a multi‐application experimentation platform developed to keep the cornea alive for prolonged periods of time in a physiological environment of intraocular pressure and environmental renewal.AimTo evaluate whether this model can be used to test the efficacy of antiviral eye drops.MethodsHSV‐1 keratitis was obtained as previously described: paired corneas discarded from our eye bank were stored in the BR for 14 days (D) at 21 mmHg and 2.6 µl/min of medium renewal (CorneaMax, Eurobio, France) to allow epithelial regeneration. A superficial linear epithelial lesion was then made in the centre, followed incubation of the epithelial surface for 1 hr in a solution of 106 Plaque‐Forming Unit/mL HSV‐1. Three days post‐infection, 0.15% ganciclovir ophthalmic gel used with active molecule at 3.75 µg/ml was instilled on the cornea inside the BR, 3 times/day during 14D and 2 times/day during 5D. The gel was rinsed after 10 min with BSS. The paired cornea was treated similarly with the vehicle (carbomer 974P). Changes in epithelial ulceration size were monitored daily by non‐invasive imaging through the transparent BR windows. Histology and immunolabeling (HSV‐1 gpB, epithelial markers) were done at D40.ResultsFor the control corneas, the entire epithelium desquamated at D10. At D40 all keratocytes expressed gpB and endothelial cells partly disappeared. For ganciclovir‐treated corneas, only half of the epithelium desquamated at D14, then it began to regenerate during the third week to reach 1 to 3 epithelial layers at D40. Epithelial and endothelial cells were not infected (gpB‐). Some keratocytes were infected only in the periphery.ConclusionsModel of herpetic keratitis in the BR has a high potential to evaluate antiviral eye drops efficacy.
Monitoring blood reflux through the trabecular micro‐bypass device “iStent inject”during gonioscopy as a way to follow the patient and optimize the therapy
Authors: Azzawi Abdelwahhab , Sachs Helmut 
Published: January 2021

PurposeA case series of monitoring Blood reflux during Gonioscopy after 3‐6 months of the implantation of the trabecular micro‐bypass device “iStent inject® or iStent Inject W®” with cataract surgery to evaluate the effectiveness of the implantation and adjust the therapy.MethodsA gonioscopy with Goldmann 3 mirror classic contact lens 903 was performed on 22 eyes after implanting iStent inject® or iStent inject w® combined with cataract surgery. Patients are diagnosed with early primary open‐angle Glaucoma (POAG) or with ocular hypertension (OHT), and they were treated with at least one hypotensive eye drops (ED) before the surgery. The patients were controlled regularly after the surgery, and the therapy was reduced in most cases and adjusted to reach the target pressure according to the guidelines of European glaucoma society with consideration of the IOP before the surgery. The Gonioscopy was performed between 3 and 6 months after the surgery to evaluate the blood reflux through the iStent.ResultsThe mean IOP in the patients after the surgery was 18 mmHg with a mean of 1.25 eye drops with no complications, and no patient needed a second surgery. The mean reduction of eye drops (ED) was 1 ED after the surgery. Three of the patients who had IOP less than 15 mmHg had blood reflux through one of the two iStents during Gonioscopy.ConclusionsGonioscopy may offer a reasonable and effective method to evaluate the function of iStent after implanting iStent with no extra costs, and it could help in adjusting the therapy after the surgery.
Immunolabelling of usual and putative limbal stem cells markers on flat mounted whole human corneas
Authors: Vaïtinadapoulé Hanielle , Perrache Chantal , Forest Fabien , Marc Dumollard Jean , Gavid Marie , Gain Philippe , Thuret Gilles , He Zhiguo 
Published: January 2021

AbstractPurposeThe permanent regeneration of the human corneal epithelium involves the proliferation of basal cells of corneal epithelium for compensating for daily desquamation (Majo et al. Nature 2008) and of limbal stem cells (LSCs, Cotsarelis, Cell 1989) located in limbal crypts that can be stimulated in case of important epithelial lesion. Many markers of LSCs had been described in the literature but some are controversial, particularly for their specificity.AimTo evaluate the specificity of the main LSC markers proposed in the literature and map their expression on the surface of the cornea.MethodsReferenced or potential markers of LCSs were tested on cross‐sections of fresh human corneas (body donation to Sciences, time between death‐retrieval‐fixation <21 hr) using a standard technique of immunohistochemistry: ABCB5, ΔNp63, P63, ABCG2, Pax6, CK15, CK17, N‐cadherin, Connexin 43, EGFR, vimentin, Sox2, OCT 4. Secondly, antibodies that showed positivity on cross sections were used for immunolabelling flat‐mounted whole cornea to map their expression.ResultsNone of 13 markers was exclusively located at the basal layer of the limbal epithelium. Nevertheless, CK15 appears to be the most promising marker because of its very strong staining in the limbal basal cell layer. However, its staining was also found, less intense, in basal layer of conjunctival epithelium and medium layers of limbal epithelium. On flat‐mounted corneas, CK15 was expressed in a circular ring corresponding to the anatomical limbus, but CK15+ cells were not restricted within the Vogt palisades (or limbal crypts on cross sections).ConclusionsA marker of cells located exclusively in limbal crypts or Vogt palisades and presumed to be the true LSCs is still missing. CK15 seems the most specific markers for limbal basal cells but not restricted to specific niches.
Prolyl oligopeptidase inhibitor KYP‐2047 alleviates inflammation in retinal pigment epithelium cells
Authors: Toppila Maija , Ranta‐aho Sofia , Bhattarai Niina , Korhonen Eveliina , Hytti Maria , Kauppinen Anu 
Published: January 2021

AbstractPurposeRetinal inflammation and dysfunctional protein clearance are characteristics of age‐related macular degeneration (AMD) – the most common cause of blindness in the developed countries. Prolyl oligopeptidase (PREP) is a serine protease that has several functions in the cells including regulation of inflammatory mediators. In this study, we investigated the effects of PREP inhibitor KYP‐2047 on inflammation in retinal pigment epithelium (RPE) cells with blocked proteasomal clearance.MethodsProteasomal degradation in ARPE‐19 cells was inhibited by MG‐132. The cells were exposed to the PREP inhibitor KYP‐2047 (1 µM) for 15 min before adding MG‐132 (5 µM) for 6 h or 24 h. Cytotoxicity was determined using the lactate dehydrogenase (LDH) assay. Cytokine levels (IL‐6 and IL‐8) were measured from cell culture medium samples using the enzyme‐linked immunosorbent assay (ELISA) method. Autophagy markers p62/SQSTM1 and LC3 were determined using Western blot technique.ResultsCells suffering from MG‐132‐induced proteasomal blockade showed higher LDH leakage and IL‐8 secretion as well as accumulation of the autophagy marker p62/SQSTM1 and increased conversion of LC3‐I to LC3‐II. The PREP inhibitor KYP‐2047 reduced the levels of LDH and pro‐inflammatory cytokines IL‐6 and IL‐8 in human RPE cells exposed to MG‐132. KYP‐2047 did not have a significant effect on the levels of autophagy markers at the 24 h time point.ConclusionsPREP inhibitor KYP‐2047 protects human RPE cells from MG‐132‐mediated cytotoxicity and alleviates inflammation. Further studies are needed to uncover the underlying mechanism, but PREP inhibition is a promising therapy target in AMD.
Endothelial viability after DSAEK cutting of corneas stored in Tissue‐C organoculture medium and deswelled in Carry‐C medium (Alchemia)
Authors: Vaïtinadapoulé Hanielle , Al‐Bourgol Samy , Ninotta Sandrine , Theilliere Christian , Gouttefange Catherine , Acquart Sophie , Thuret Gilles , Gain Philippe 
Published: January 2021

AbstractPurposeIn France, the adoption by an eyebank of a new technique for graft preparation and/or a new CE‐marked storage medium requires the submission of a “process” file to the competent health authority. This file must include pre‐clinical data on corneas stored in this medium or prepared with the new technique.AimTo analyze the endothelial alterations induced by the pre‐cutting with the microkeratome of corneas stored in Tissue‐C organoculture medium and deswelled in Carry‐C medium.MethodsTen pairs of corneas (donation for scientific purposes) were retrieved by in situ corneo‐scleral cutting and immersed in 100mL of Tissue‐C (Alchemia, Italy) and stored at 31°C in a dry incubator for 30 days (D). At the beginning (D2 to D5) and end (D30) of storage, the usual quality control was performed: central endothelial cell density (ECD) after incubation with 0.9% NaCl, transparency, thickness (OCT) and microbiological control. At D30, the corneas were transferred to a deswelling Dextran‐containing medium (Carry‐C, Alchemia) for 24 H. After randomization, one cornea of each pair was pre‐cut with the MORIA linear microkeratome (DSAEK, single pass, objective #150 µm) and put back in Carry‐C for 72H. The endothelial viability (viable ECD) of the all corneas was then measured by triple Hoechst‐Ethidium‐Calcein‐AM staining as we previously described (IOVS2011, Cornea2014).ResultsThe initial ECD was 2401 ± 815 for the DSAEK group and 2298 ± 805 cells/mm2 for the control group (p = 0.386) and at D30, respectively, from 1945 ± 587 versus 1868 ± 577 cells/mm2 (p = 0.421). One cornea was perforated. Graft thickness was 187 ± 112 um. For the other 9 pairs, the viable ECD (by definition always lower than bank ECD) was 1061 ± 448 for the DSAEKs and 1149 ± 409 cells/mm2 for the controls (p = 0.066). The pre‐cut grafts had fewer endothelial folds.ConclusionsPre‐cutting by bank of corneas stored in Tissue‐C/Carry‐C does not induce significant additional cell loss compared to control.
Automated macular segmentation in compressive optic neuropathy and glaucomatous optic neuropathy
Authors: Machado Soares Ricardo , Braga Joana , Da Silva Fernandes Joana , Alves Hélio , Ribeiro Lígia , Meira Dália 
Published: January 2021

PurposeCompressive optic neuropathy (CON) can sometimes mimic glaucomatous optic neuropathy (GON), leading to inadequate management. Our purpose was to compare the macular area of patients with CON and GON to healthy controls (HC) and to determine if the macula area was able to distinguish between CON and GON.MethodsRetrospective, observational, cross‐sectional study, enrolling 70 patients (22 patients with CON, 25 with GON and 23 HC). Subjects were matched by age (median of 61 [±11.6]), and one eye was randomly selected. Spectral‐domain optical coherence tomography (SD‐OCT) images were generated using the Spectralis® OCT device (Heidelberg, Germany). An automated algorithm was used to segment the macular retina into nine layers and evaluate the thickness of each layer in the foveal, inner, and outer Early Treatment Diabetic Retinopathy Study (ETDRS) subfield rings. Data were compared across all study groups using Kruskal‐Wallis test, followed by pairwise comparisons for each duo. The resulting p‐values were adjusted using the Bonferroni correction and significance level was set at 0.05.ResultsHC had thicker inner retinal layer (IRL) compared with GON and CON (p < 0.05). Global retinal thickness was significantly thinner in the inner superior (IS) and inner nasal (IN) subfields in CON compared with GON (p < 0.05). The IRL is thinner in CON, specifically in the inner and outer nasal subfields, compared to GON. The IN subfield was thinner in the retinal nerve fibre, ganglion cell and inner plexiform layers (IPL) in CON compared with GON (p < 0.05). No differences were found on the inner nuclear, outer plexiform, outer nuclear and outer retinal layers between groups.ConclusionsOur findings suggest that IRL is thinner in both CON and GON, and the IN subfield is specially affected in CON compared with GON. Despite not being our primary outcome, our results suggest that, compared to HC, layer neurodegeneration in both CON and GON affects mainly the inner layers and that does not extend to layers external to IPL. In conclusion, evaluation of the macular segmented layers damage, by SD‐OCT, may be a useful add‐on tool in the differential diagnosis between these two entities, when their manifestations overlap.
Visual impairment and associated risk factors among school‐age children in Kashatagh, Artsakh
Authors: Giloyan Aida , Harutyunyan Tsovinar , Petrosyan Varduhi 
Published: January 2021

AbstractPurposeVisual impairment in childhood can have a profound impact on child's development, restricting social life and limiting physical and educational opportunities. This study assessed the prevalence of visual impairment and associated risk factors among school‐age children living in Kashatagh, Artsakh.MethodsThe researchers recruited 454 school‐age children from seven urban/rural schools in 2016. The structured questionnaire included questions about socio‐demographics, frequency of visiting ophthalmologist, ocular health and vision acuity. All children underwent a detailed ophthalmic examination including measurement of visual acuity, cycloplegic refraction and dilated eye fundus examination. Visual impairment was defined by a presenting visual acuity <6/12 with or without correction in the better eye. Descriptive statistics and logistic regression were used to analyze the data.ResultsChildren's mean age was 11.1 (3.28), ranging from 6 to 19 years. About 55% participants were girls. The majority of children never visited ophthalmologist (86.1%). The prevalence of presenting visual impairment without correction and with available correction in the better eye were 6.0% and 4.6%, respectively. After best correction, all visually impaired cases improved their vision to ≥6/12. The majority of visually impaired participants had myopia (55.6%). In the adjusted analysis, age (OR = 1.22; 95% CI: 1.04–1.44), female gender (OR = 3.33; 95% CI: 1.07–10.3) and frequency of visiting an ophthalmologist (OR = 11.8; 95% CI: 4.35–32.2) were associated with presenting visual impairment without correction. Visually impaired children more frequently visited an ophthalmologist compared to those with no visual impairment.ConclusionsCorrectable visual impairment is a major problem in this sample. All visually impaired cases improved their vision to ≥6/12 after best correction in the better eye. School‐based vision screening programs and provision of spectacles can decrease visual impairment in school‐age children.
Epidemiological analysis of conjunctival lymphoma: a population‐based study, 2000–2016
Authors: Mikhael Sandra , Tadrosse Abanoob , Yassa Arsany , Mikhael Mina , Eloy Jean‐Anderson 
Published: January 2021

PurposeTo study epidemiological trends of conjunctival lymphoma.MethodsA retrospective, population‐based analysis of patient data extracted from the Surveillance, Epidemiology, and End Results Registry US database was conducted from 2000 to 2016. Incidence (IR) was calculated in number of cases/million/year. Disease‐specific survival (DSS) was calculated using the Kaplan‐Meier method.Results978 cases of conjunctival lymphoma (CL) were identified, of whom 52% were females and 65% were white. The most common histological subtypes were mucosa‐associated lymphoid tissue (MALT; 70.9%) and follicular lymphoma (11.7%). Diffuse large B‐cell lymphoma constituted only 3.4% of cases. Overall IR of CL was 0.7. Annual percent change showed a significant decline (−1.4) in IR of CL during 2000–2016 (p = 0.02). There was no difference in IR between sexes (p = 0.34). IR in Asian/Pacific Islander patients (0.9) was significantly higher than that in white (0.6) and black (0.5) [p < 0.01] patients. Mean diagnostic age in years was 59.3 [Standard Deviation = 17.9]. IR in patients older than 65 years old (y/o) [2.36] was significantly higher than that in those 22–64 y/o (0.63; p < 0.01) and those younger than 21 y/o (0.06, p < 0.01). IR of unilateral cases (0.6) was significantly higher than that of bilateral ones (0.1) [p < 0.01] and there was no significant difference between left and right laterality (p = 0.72). 5‐year DSS during 2000–2012 was 97.4%. Race and sex did not affect survival. DSS was similar during 2010–2012 (98.5%) and 2000‐02 (94.1%) [p = 0.099].ConclusionsConjunctival lymphoma incidence is highest in the elderly and Asian/Pacific Islander populations and is not affected by sex. Incidence is decreasing over the timeframe 2000‐2016. Most common histology for CL is MALT. Overall 5‐year DSS for CL is 97.4%, and it is not affected by sex or race.BibliographySurveillance Research Program, National Cancer Institute SEER*Stat software (seer.cancer.gov/seerstat) version 8.3.6.