bluuebunny/crossref_metadata_2025_split huggingface dataset:
SMAD2/3 activation and myofibroblast conversion of corneal fibroblasts is fully dependent on the activation of MELK by the noncanonical TGFβ1 pathway
Authors: Wolosin Jose , Wang Zheng , Bernstein Audrey 
Published: December 2022

AbstractPurpose: To identify the role of noncanonical responses to TGFβ on the early canonical response and the subsequent fibroblasts into myofibroblasts conversion (FMC).Methods: Human corneal fibroblasts were seeded in a serum‐free medium for 48 h. TGFβ1 (TGFβ‐2 n/ml) was added and cultures were lysed 1–15 min later. Kinase inhibitors were added 10 min beforehand. Identical amounts of soluble protein were then used for Western blot, to measure the level of expression of p.SMAD2 (S465/467), p.SMAD3(S423/425), p.TAK1 (S412), p.JNK1/2 (JNK;T183/185), p.p38 (T180/182), p.MELK(T167/171), α‐SMA, and GAPDH. MELK esiRNA was electroporated 48 h prior to the addition of TGFβ. FMC was assessed by α‐SMA immunohistology and morphology.Results: In the corneal fibroblasts, the TGFβ‐induced SMAD2/3 phosphorylation (canonical response), displayed a 6–8 min latency period that was followed by rapid, p.SMAD2/3 accumulation to peak response within 10 min. Overt de novo expression of α‐SMA and morphological FMC was observed after 48 h. Inhibition of MELK, a leucine zipper kinase, by OTSS167 or MELK8A, or its downregulation by esiRNA, blocked both responses. In contrast to the delayed canonical response, phosphorylation of TAK1 (noncanonical response), reached its peak within 1 min, as did JNK and p38, TAK1 downstream targets in the mitogen‐activated kinase cascade. The temporal difference was exploited to examine the relationship between the two responses. Inhibition of TAK1 by TAKinib or 5Z‐7‐oxozeaenol blocked the phosphorylation of JNK, p38, MELK, and SMAD2/3 and the FMC. In turn, inhibition of JNK by SP600125 blocked the phosphorylation of MELK and SMAD2/3; p38 inhibition had no effect. MELK inhibition did not inhibit the phosphorylation of either TAK1 or JNK. Finally, phosphorylation of JNK and MELK via TNFα or LPS, did not induce p.SMAD2/3 or blocked the effect of TGFβ on SMAD. These results are consistent with the event sequence, TGFβ → p.TAK → p.JNK1/2 → p.MELK + TGFβ → p.SMAD2/3.Conclusions: SMAD2/3 activation is fully dependent on TAK1 activity, challenging the notion of separated canonical ‐ noncanonical responses. MELK, a previously unknown component of the response to TGFβ, plays a central role in SMAD phosphorylation. The molecular interaction between the TGFβ receptor, MELK, and SMAD2/3 remains to be elucidated.
Swept‐source optical coherence tomography angiography in toxoplasmic retinochoroiditis
Authors: Doukh Meryem , Ksiaa Imen , Amor Hager Ben , Abroug Nesrine , Jelliti Bechir , Khochteli Sana , Khairallah Moncef 
Published: December 2022

AbstractPurpose: To describe optical coherence tomography angiography (OCTA) findings in eyes with active and scarred toxoplasmic retinochoroiditis.Methods: OCTA scans in active (25 eyes) and scarred (17 eyes) retinochoroiditis were retrospectively reviewed at Fattouma Bourguiba University Hospital, Monastir, Tunisia.Results: In active lesions, OCTA findings included a non‐detectable flow signal area in retinal vascular plexuses and choriocapillaris in all 25 eyes (100%), an abnormal intraretinal vascular process in two eyes (8%), and an associated area of retinal flow deficit secondary to branch retinal artery occlusion in one eye (4%). In scarred lesions, OCTA findings included a flow deficit area in retinal vascular plexuses and choriocapillaris in all 17 eyes (100%), a visibility of larger deeper choroidal vessels at the level of choriocapillaris in nine eyes (53%), and a well‐defined intraretinal vascular network in one eye (5.9%). Peripapillary scars were associated on OCTA with wedge‐shaped loss of radial peripapillary capillaries with corresponding localized retinal nerve fibre layer defect and visual field loss.Conclusions: OCTA allows to non‐invasively detect retinal and choroidal vascular changes in active and scarred toxoplasmic retinochoroiditis.References1. Perez AL, Lozada RA, Emanuelli A, Oliver AL. Optical coherence tomography angiography findings in macular toxoplasma retinochoroiditis: A case report. Am J Ophthalmol Case Rep. 2020;19:100764.2. Atas F, Kaya M, Toprak T, Akbulut Yagci B, Selver A, Saatci AO. Measurement of the active toxoplasma retinochoroiditis lesion size during the disease course with swept‐source optical coherence tomography angiography: A retrospective image analysis. Int Ophthalmol. 2021;41(12):4127–4135.
Effect of photobiomodulation in suppression of oxidative stress on retinal pigment epithelium
Authors: Lee Yeojin , Kim Jongmin , Won Jae Yon 
Published: December 2022

AbstractPurpose: As the world undergoes aging, the number of age‐related diseases has increased. One of them is disease related to retinal pigment epithelium (RPE) degeneration, such as age‐related macular degeneration, causing vision loss without physical damage in the ocular system. It is the leading cause of blindness, with no cure. Although the exact pathogenesis is still unknown, the research shows that oxidative stress is one of the risk factors. Various molecules have been reported as anti‐oxidative materials; however, the disease has not yet been conquered. Here, we would like to introduce photobiomodulation (PBM). PBM is a non‐invasive treatment based on red and near‐infrared light and has been used to cure various diseases by regulating cellular functions.Methods: The BM‐ECM was prepared and the RPE cell line was cultured. The hypoxia condition was induced via cobalt chloride treatment. The barrier function was evaluated via ZO‐1, a marker of tight junction, expression, and Transepithelial Electrical Resistance (TEER) measurement. The clearance function was evaluated via MERTK expression and digestion of polystyrene beads. The visual function of RPE was examined via expression of RPE65, the enzyme for phototransduction. The oxidative stress was compared via ROS generation.Results: In this study, we confirm the antioxidant effect of PBM in retinal pigment epithelium via an RPE model with hypoxia. The function of RPE is protected by PBM against damage from hypoxia. Furthermore, we observed the protective mechanism of PBM by its suppression effect on reactive oxygen species generation.Conclusions: These results indicate that PBM shows great potential to cure RPE degeneration to help patients with blindness.
Assessment of compliance with current driving legislation in Spanish patients with visual disturbances
Authors: Franco Pedro Esteban Quílez , de Imperial Ollero Juan Antonio Miralles , Alacid Alberto López , Navarro Inmaculada Sellés 
Published: December 2022

AbstractPurpose: To determine whether patients who do not fulfil some of the requirements of current Spanish driving regulation but still have their driving licence have stopped driving. Furthermore, to assess the extent to which ophthalmologists know such legislation and whether they counsel their patients accordingly.Methods: Two surveys were conducted. For the first one, we contacted 133 patients who did not fulfil either current visual acuity (VA) or visual field requirements. We asked them to answer 10 questions about their driving habits and their subjective perception of visual quality and road safety. For the second one, we contacted 137 ophthalmologists and presented them with 12 questions: 6 of them inquired about current legislation requirements and the rest of them looked at their opinion about the importance of vision in driving matters.Results: 65% of patients (95% CI: [57%–74%]) kept driving despite not fulfilling minimal legal requirements. Continuation of driving was associated with having better VA (p < 0.005), better subjective perception of visual quality (p < 0.0000001), having had a previous conversation with their ophthalmologist about driving (p < 0.05; OR = 3.50) and not being pressured by their families for driving cessation (p < 0.05, OR = 2.67). Regression models showed that subjective perception of visual quality was not a good predictor of objective VA neither among non‐drivers (R2 = 0.228) nor drivers (R2 = 0.072). Most ophthalmologists (72.3%) correctly answered no more than 3 out of 6 questions about the requirements of current legislation. Only 7 (5.1%) had received specific training about these matters. Most of them (97.1%) agreed that sufficient visual quality is of great importance for driving safely.Conclusions: Most drivers who keep their driving licence despite falling below the minimum legal requirements to drive continue driving. Current evaluations for licence renewal could be insufficient. Family pressure seems to be important in driving cessation. Ophthalmologists play a role in patients' counsel about driving, but their knowledge seems to be limited. New training initiatives could be beneficial.
Are choroidal vascularity parameters influenced by age, hypertension and visual acuity in pachychoroid spectrum diseases?
Authors: Abid Yosra , Sassi Hela , Ouederni Meriem , Nouri Slim , Chaabouni Karim , Cheour Monia 
Published: December 2022

AbstractPurpose: To identify correlations of subfoveal choroidal thickness (SFCT) and choroidal vascularity index (CVI) with age and best corrected visual acuity in eyes with pachychoroid spectrum diseases (PSD) and their association to systemic hypertension using a retrospective study design.Methods: Our study included 82 eyes of 45 patients with PSD: 15 eyes had uncomplicated pachychoroid (UCP), 27 had pachychoroid pigment epitheliopathy (PPE), 32 had central serous chorioretinopathy (CSC) and 8 had pachychoroid neovasculopathy (PNV). All patients underwent DRI TRITON Swept‐Source Optical Coherence Tomography (SS‐OCT). A 1500 mm width subfoveal choroidal area was defined in SS‐OCT B‐scans and divided into luminal area (LA) and stromal area (SA) by the imageJ binarization technique. CVI was defined as the ratio of LA to the total subfoveal choroidal area (TCA).Results: The mean SFCT was 393, 23 ± 95, 65 μm and it was correlated negatively with age (p = 0.003, R = −0.326). We found a negative correlation of TCA and LA with age (p = 0.043, r = −0.227, p = 0.04, r = −0.230). Mean choroidal vascularity index was 64.45 ± 2, 74% with no correlation with age (p = 0.169, R = −0.155). No parameter among SFCT, CVI, TCA, LA and SA had a significant correlation with BCVA or spherical equivalent. Patients with essential hypertension were found to have significantly thinner SFCT when compared with healthy controls (374, 49 ± 83 141 μm vs. 423, 98 ± 130 466 μm, p = 0.046). CVI was not affected by a history of systemic hypertension (64.05 ± 1.97 vs. 64.83 ± 3.3, p = 0.207). The total choroidal area, luminal area, and stromal area, as well as the CVI correlated with SFCT (p < 0.001, r = 0.817, p < 0.001, r = 0.821, p < 0.001, r = 0.750 and p < 0.001, r = 0.371 respectively).Conclusions: SFCT and CVI are dynamic parameters that are affected in patients with pachychoroid diseases. Unlike CVI, SFCT is also affected by age, ocular and systemic factors like hypertension. CVI may be a more specific biomarker for PSD.
Administration of minocycline reduces microglia and Caspase‐3 activation but does not mitigate retinal ganglion cell loss after ocular hypertension in mice
Authors: Valiente‐Soriano Francisco Javier , Sánchez‐Migallón María Cielo , Di Pierdomenico Johnny , Ortega Alejandro Gallego , García‐Ayuso Diego , Vidal‐Sanz Manuel , Agudo‐Barriuso Marta 
Published: December 2022

AbstractPurpose: To study the effect of minocycline administration on the evolution of retinal ganglion cell (RGC) loss, the number of microglial cells and the activation of the apoptotic signal Caspase‐3 after ocular hypertension (OHT) in mice.Methods: In left eyes of Swiss albino mice, OHT was induced by diode laser photocoagulation of the limbal and episcleral veins and the intraocular pressure (IOP) evolution was monitored with a TonoLab®. Mice received daily intraperitoneal injections of minocycline hydrochloride (45 mg/kg) diluted in saline starting the day before OHT induction until the day of sacrifice (4 or 15 days, n = 12–14 per group/time point). Control groups received saline injections. Retinas were divided into two groups. Retinas of the first group were double immunodetected against Brn3a and active Caspase‐3 to identify surviving (Brn3a+RGCs) and apoptotic (a‐Casp3+RGCs) RGCs. In the second group, RGCs were retrogradely traced with Fluorogold 7 days before OHT induction and retinas were immunodetected against Iba‐1. Total Brn3a+RGCs, a‐Casp3+RGCs, Iba‐1+cells and FG+microglial cells in the RGC layer were manually or automatically quantified, and the distributions were analysed by topographical maps.Results: IOP was significantly elevated 24 hours after OHT induction (33.2 ± 6.1 mmHg) and returned to baseline at 1 week (17.3 ± 5.3 mmHg). This evolution was not altered by minocycline administration. Although minocycline treatment was not successful in protecting Brn3a+RGCs at 4 (40 878 ± 4860 vs 43 526 ± 2714) nor at 15 (16 161 ± 12 650 vs 12 919 ± 6085) days after OHT, the numbers of a‐Casp3+RGCs, Iba‐1+cells and FG+microglial cells were significantly lower in the minocycline‐treated groups at both 4 (58.6%, 46.8% and 24.5%, respectively) and 15 (33.6%, 54.2% and 37.3%, respectively) days compared to the saline‐treated groups (100%).Conclusions: Minocycline treatment decreases the number of microglial cells and caspase‐3 activation but does not reduce RGC loss after OHT.
Report of two cases of subconjunctival haemorrhage as first sign of post‐pregnancy acquired haemophilia a (AHA)
Authors: Fruschelli Mario , Marini Davide , Luciani Diego , Caroni Federico , Puccetti Luca 
Published: December 2022

AbstractPurpose: To show that subconjunctival haemorrhage may be the first sign of post‐pregnancy acquired A haemophilia (AHA), a potentially fatal bleeding disorder.Methods: Case reports.Results: Two thirty‐year‐old caucasic women at first pregnancy presented slight pain without visual loss 2 weeks after uncomplicated spontaneous full‐term delivery. The only clinical findings were a subconjunctival bleeding for both and a small haematoma in the first case. The clinical history was negative for spontaneous bleeding also in infantile age and at previous surgery and no bleeding tendency was observed in the main relatives. The first patient was lost on follow‐up and admitted to urgent haemathologic evaluation due to large multiple cutaneous hematomas, a massive deep muscle haematoma, asthenia and dyspnea. In the second case quick routine coagulation labs were performed immediately after the subconjunctival bleeding. In both cases PTT ratio was increased. An extensive study of blood coagulation suggested the presence of a circulating inhibitor against factor VIII [1,2], confirming the diagnosis of AHA. First case was treated with a bypassing agent, however the eradication was obtained only with immunosuppressive therapy. Second case was initially placed in close follow‐up, since the post‐pregnancy form is often self‐limiting, however an immunosuppressive therapy was set after clinical progression due to the appearance of spontaneous hematoma.Conclusions: AHA is a relevant acquired bleeding disorder that can be fatal if not quickly recognized and treated even when the initial clinical signs are minor [3], how it can happen in post‐pregancy mainly in the first month [4]. Since it may present as spontaneous subconjunctival haemorrhage, routine hemathological and coagulation labs should be performed to exclude this serious condition or once identified, allow for proper clinical management.
Osmoprotective and bioprotective effect of threalose 3% under desiccating conditions: An in vitro study
Authors: del Papa Melina Sol , Passerini María Silvia , Berra Alejandro 
Published: December 2022

AbstractPurpose: To study the osmoprotective and bioprotective effect of trehalose 3% during desiccation conditions.Methods: Normal human conjunctival epithelial cells (IOBA‐NHC) were exposed to: 1) culture media DMEM‐F12 (Control); 2) Carboxymethylcellulose 0.5% + glycerin 0.9% + PEG 400 0.25% (Vehicle); 3) Vehicle + trehalose 3% (Trehalose). Cultures were treated for 1 h and then exposed to desiccating conditions (25°C and 36–38% relative humidity). Metabolic activity by MTT assay and cell viability by Triptan Blue assay was evaluated. To evaluate water retention capacity, a gravimetric test was carried out by soaking a filter paper with 5 ml of each solution under evaluation and recording its weight over time. The data were analysed by analysis of variance (ANOVA) with a post hoc Tukey multiple comparisons test.Results: MTT assay: During the first 30 min under desiccating conditions there were no differences between groups (Control: 0.586 ± 0.012; Vehicle: 0.575 ± 0.278; Trehalose: 0.523 ± 0.035). After 45 min of exposure metabolic activity sharply decreased in control group (0.111 ± 0.007) and vehicle group (0.107 ± 0.027), while trehalose group maintained metabolic activity values (0.560 ± 0.008; p < 0.001).Triptan Blue assay: The cell viability of the trehalose group was maintained throughout the time (10 min: 62.7 ± 2.5; 15 min: 58.7 ± 8.1; 20 min: 62.3 ± 10.8; 30 min:41.7 ± 8.5%) and was statistically superior to the control and vehicle groups at all evaluated times (p < 0.01).Water retention capacity: Water content of papers soaked with control and vehicle solution was almost completely evaporated (less than 20% of water retention) at 10 min on desiccating conditions, while trehalose group show a significant delay of water evaporation of 5 and 7 min regarding the vehicle and control group, respectively.Conclusions: The addition of trehalose 3% prolonged cell viability and extended water retention time in conditions of high environmental dryness. This study confirms the bioprotective and osmoprotective role of trehalose evaluated in previous investigations and provides scientific evidence of its properties, representing an additional benefit in the treatment of dry eye.
Gene editing strategy for development of an inherited retinal dystrophy model based on iPSC‐derived photoreceptors with mutation in the PROM1 gene
Authors: Fernandez‐Bueno Ivan , Puertas‐Neyra Kevin , Hernández‐Rodríguez Leticia , Gobelli Dino , Simarro Maria , Usategui‐Martín Ricardo , Coco Rosa , de la Fuente Miguel Angel 
Published: December 2022

AbstractPurpose: The PROM1 gene encodes the protein Prominin‐1 which plays a critical role in the morphogenesis of photoreceptors outer segments. Mutations in the PROM1 gene are related to Inherited Retinal Dystrophy (IRD) phenotypes. The c.1354dupT mutation in the PROM1 gene causes a premature stop codon. The aim of the study was to define the more accurate gene editing strategy for development of an IRD model based on iPSC‐derived photoreceptors.Methods: Firstly, the most accurate CRISPR/Cas9 machinery was determined by the web tool for genome editing CRISPOR and the DNAStar Lasergene software. gRNAs were selected, accordingly to their proximity to the mutation position and their specificity and efficiency scores. Then, silent mutations were added to the repair oligonucleotides (Alt‐R HDR Donor Oligos), by using the web tool MolBiotools, to generate a restriction site for the Ssp1enzyme. Afterwards, to confirm the in silico information, in vitro studies were performed. The U2OS cell line and the control iPSC line ([FiPS] Ctrl1‐Ep6F‐5) were transfected with each selected gRNA, their respective repair oligonucleotide, and the endonuclease Cas9‐protein, by using the Neon Transfection System. Finally, a digestion with Ssp1 were performed to specifically detect the edited genomes.Results: Several gRNAs were selected in silico. In vitro studies showed higher genome editing efficiency in two of them (40RE and 47RE). Finally, the gRNA guide 40RE and its respective repair oligonucleotide were selected.Conclusions: In‐silico and in‐vitro studies concluded that the gRNA guide 40RE was the most accurate for development of an in vitro IRD model based on iPSC‐derived photoreceptors with a mutation in the PROM1 gene. Gene editing strategy designed in silico should be validated in vitro before transfecting the target cells.
Activation of Nrf2‐mediated protective pathways by nature‐inspired hybrids improves proteostasis in retinal pigment epithelial cells
Authors: Koskela Ali , Manai Federico , Basagni Filippo , Liukkonen Mikko , Rosini Michela , Govoni Stefano , Monte Massimo Dal , Smedowski Adrian , Kaarniranta Kai , Amadio Marialaura 
Published: December 2022

AbstractReferences: Antioxidant systems are essential in many elderly diseases, including age‐related macular degeneration (AMD). Oxidative stress, inflammation, protein degradation impairment, are well‐described in AMD retinal pigment epithelial (RPE) cells. Nrf2 (NF‐E2‐related factor 2), master regulator of antioxidant defence, is linked to AMD, autophagy, inflammation. We previously showed that nature‐inspired hybrids (NIH1‐3) induce Nrf2‐mediated protection against AMD‐related pro‐oxidant stimuli in RPE cells.Purpose: In human RPE cells we aimed to further investigate NIH1‐3 for capability to modulate Nrf2 pathway and provide cytoprotection against stress caused by dysfunction of protein homeostasis.Methods: ARPE‐19 cells were treated with 5 μM NIH 1–3, NIH4 (the derivative unable to activate Nrf2) or vehicle (dimethyl sulfoxide; control) for a minimum of 3 h. Proteasome inhibitor MG‐132 (5 μM) and autophagy inhibitor Bafilomycin‐A1 (50 nM) were added after 24 h NIH pre‐treatment. Levels of Nrf2, its target HO‐1 and SQSTM1/p62, the autophagy flux marker LC3‐II, were analysed by Western blotting, whilst IL‐8 release by ELISA, respectively. Cell survival was also assessed.Results: NIH1‐3 activate Nrf2‐pathway and increase HO‐1 and SQSTM1/p62 expression, in turn exerting beneficial effects on intracellular redox balance, without modification of the autophagy flux. NIH1‐3 treatments predispose ARPE‐19 cells to a better response to the following exposure to proteasome and autophagy inhibitors, as revealed by the higher cell survival and decreased secretion of the pro‐inflammatory IL‐8 compared to NIH‐untreated cells. Interestingly, also NIH4 compound, through an Nrf2‐independent pathway, exerts pro‐survival and anti‐inflammatory effects, although to a less extent than NIH1‐3.Conclusions: Our findings suggest that all NIHs are interesting for their cytoprotective properties and confirm Nrf2 as a valuable pharmacological target in contexts characterized by oxidative stress.
Ocular biometry in adults from a population‐based study in Germany
Authors: Michael Ralph , Wirkner Kerstin , Engel Christoph , Loeffler Markus , Kirsten Toralf , Rauscher Franziska 
Published: December 2022

AbstractPurpose: To obtain normal ocular biometry data in healthy European subjects.Methods: As part of the large population‐based LIFE‐Adult study (Leipzig Research Centre for Civilization Diseases), randomly selected participants from Leipzig, Germany were evaluated with the ZEISS IOL‐Master 700 (Carl Zeiss Meditec, Jena, Germany). Age range was 25–85 years, with 50% of participants above 70 years by study design and mean age of 64 years. Axial length (AL), central corneal thickness (CCT), aqueous depth (AQD), lens thickness (LT) and corneal radii (R1 & R2) were assessed in 1365 right eyes (two mandatory repeated measurements, with quality check passed).Results: Mean AL was 23.53 mm and mean CCT was 558 micrometres with no significant change with age. AQD decreased from 2.61 mm at 50 years to 2.44 mm at 80 years. LT increased from 4.36 mm at 50 years to 4.50 mm at 80 years. LT was 4.69 mm in eyes with AL of 22 mm and 4.38 in eyes with AL of 25 mm. Mean R1 was 7.81 mm and mean R2 7.66 mm with no significant change with age.Conclusions: Above 25 years of age, we found no difference of axial length or corneal thickness with age. The lens was found continuously thicker with age and aqueous depth decreased with age. Short eyes had thicker lenses compared to long eyes; probably an adaptation to maintain a sharp retinal image.