bluuebunny/crossref_metadata_2025_split huggingface dataset:
How long should urologists observe patients with prostate cancer after radical prostatectomy? Weibull analysis
Authors: Hasegawa Masanori , Yamada Koichiro , Shigeta Keisuke , Yuzuriha Soichiro , Kawakami Masayoshi , Kim Hakushi , Nakano Mayura , Nitta Masahiro , Kawamura Yoshiaki , Shoji Sunao , Miyajima Akira 
Published: January 2022

ObjectivesThere is no recommended observation time for patients who have undergone radical prostatectomy for prostate cancer. This study was undertaken to determine the postoperative observation time by investigating the hazard rate for prostate‐specific antigen failure and other‐cause death using Weibull analysis.MethodsWe included 612 patients who underwent laparoscopic radical prostatectomy for localized prostate cancer between June 2002 and December 2017. Risk classification was categorized by the D’Amico risk classification, and the patients were divided into three age groups: <60, 60–69 and ≥70 years. The hazard rates at each point were derived using Weibull analysis. The optimal observation time after laparoscopic radical prostatectomy was determined as the intersection point at which the hazard rate of other‐cause death overtakes the hazard rate of prostate‐specific antigen failure.ResultsIn all groups classified by age, the hazard rate of other‐cause deaths increased over time. In contrast, the hazard rate of prostate‐specific antigen failure decreased gradually. The ≥70 years age group showed the highest hazard rate. The hazard rate of prostate‐specific antigen failure was highest in the high‐risk group. The patients aged ≥70 and 60–69 years in the low‐risk group were recommended 6 years 6 months and 14 years 8 months, respectively, for observation. The remaining patients were recommended >25 years of postsurgical observation.ConclusionsThe observation time after laparoscopic radical prostatectomy could be estimated by comparing the estimated hazard rates of prostate‐specific antigen failure and other‐cause death based on Weibull analysis. Urologists should pay attention to age and risk classifications for optimal postoperative observation.
Prevention of retinal ganglion cell death by GPR81 agonist treatment
Authors: Vohra Rupali , Tams Anna Luna Mølgaard , Morello Berta Sanz , Mouhammad Zaynab Ahmad , Hannibal Jens , Bergersen Linda Hildegaard , Kolko Miriam 
Published: January 2022

AbstractPurposeRetinal ganglion cell (RGC) death is a common characteristic for ocular neurodegenerative diseases such as glaucoma and optic neuropathies. Recently, GPR81 agonist treatment has been identified as a key modulator of mitochondrial function and cell survival. Thus, we aimed to test whether GPR81 agonist treatment likewise promotes RGC survival and energy metabolism in retinal explants and wholemounts from mice.MethodsRetinal explants were treated with 5 mM of the GPR81 agonist, 3,5‐DHBA, for two, four, 24 & 72 hours, and compared to conditions with no treatment. Additionally, explants were also treated with 15 mM of L‐glutamate to induce toxicity and simultaneously treated with GPR81 agonist. Tissue survival was assessed through lactate dehydrogenase (LDH) viability assays. Retinal ganglion cell survival was measured in murine wholemount retinas through immunohistochemical staining (IHC). Total ATP levels were quantified through bioluminescence assays.ResultsGPR81 agonist treatment increased retinal explant survival after 24 and 72 hours of exposure. No significant effect was seen in retinal explants survival after GPR81 agonist treatment alone for two hours. However, during glutamate‐induced death, supplemented GPR81 agonist treatment increased survival compared to conditions with glutamate toxicity. IHC analysis revealed that GPR81 agonist treatment for two hours promoted RGC survival in retinal wholemounts compared with no treatment. GPR81 agonist treatment also enhanced ATP levels in retinal explants after two, 24, and 72 hr of exposure.ConclusionsThe present study reveals that GPR81 agonist treatment has a neuroprotective effect on specifically RGCs and on glutamate‐induced retinal degeneration. Hence, GPR81 agonist administration may be a potential new strategy to sustain RGCs, ultimately preventing visual disability as a consequence of RGC death.
Pinosylvin extract Retinari™ sustains electrophysiological function, prevents thinning of retina and enhances cellular response to oxidative stress in NFE2L2 knock‐out mice
Authors: Tamminen Toni , Koskela Ali , Toropainen Elisa , Gurubaran Iswariyaraja Sridevi , Winiarczyk Mateusz , Liukkonen Mikko , Paterno Jussi J. , Lackman Petri , Sadeghi Amir , Viiri Johanna , Hyttinen Juha M. T. , Koskelainen Ari , Kaarniranta Kai 
Published: January 2022

AbstractPurposeChronic oxidative stress eventually leads to protein aggregation in combination with impaired autophagy, which has been observed in age‐related macular degeneration (AMD). We have recently developed a dry AMD mice model with a nuclear factor‐erythroid 2‐related factor‐2 (NFE2L2) knock‐out (KO). Pinosylvin, a polyphenol abundant in bark waste, improves retinal pigment epithelial cell (RPE) cell viability in vitro. Therefore, one can estimate that pinosylvin slow down the progression of dry AMD.MethodsWe investigated the effects of commercial natural pinosylvin extract, Retinari™, on the electroretinogram, optical coherence tomogram, autophagic activity, antioxidant capacity, and inflammation markers. Wild type (WT) and NFE2L2 KO mice were raised in 12:12 until the age of 14.8 ± 3.8 months. They were fed with either regular or Retinari™ chow (50–250 mg/kg/day of pinosylvin) for 10 weeks before the assays.ResultsRetinari™ treatment preserved significant retinal function in scotopic and photopic ERG and prevented thinning of the retina in the NFE2L2 KO mice. The NFE2L2 KO mice showed reduced ubiquitin‐tagged protein accumulation in addition to upregulation of the antioxidant enzymes superoxide dismutase 1 and catalase. Furthermore, the treatment seemed to reduce inflammation in NFE2L2 KO mice by causing a climb in the concentration of complement factor H.ConclusionsOur results demonstrate a strong protective effect of Retinari™ on the AMD disease model. We believe that pinosylvin in the Retinari™ elicits the positive effects. Thus, pinosylvin could potentially lower the risk of AMD onset and slow down its progression for which clinical trials with the compound are highly anticipated.
Oxidative stress and mitochondrial damage evoke complement component C5 activation independent of component C3 in dry age‐related macular degeneration like NFE2L2/ PGC‐1α ‐/‐ mouse model
Authors: Gurubaran Iswariyaraja Sridevi , Heloterä Hanna , Marry Stephen , Koskela Ali , Hyttinen Juha M. T. , Paterno Jussi J. , Urtti Arto , Chen Mei , Xu Heping , Kauppinen Anu , Kaarniranta Kai 
Published: January 2022

AbstractPurposeAging‐associated chronic oxidative stress and inflammation are known to be involved in various diseases, e.g. age‐related macular degeneration (AMD). Previously, we reported elevated oxidative stress, dysfunctional mitophagy and the accumulation of detrimental oxidized materials in the pigment epithelial (RPE) cells of dry AMD‐like nuclear factor erythroid 2‐related factor 2, and peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (NFE2L2/PGC1α) in a double knockout (dKO) mouse model. In this study, we show that mitochondrial‐derived oxidative stress leads to a buildup of complement component C5a, whereas the complement component C3a remains unaffected.MethodsImmunohistochemical stained retina samples from one‐year‐old NRF2/PGC1α ‐/‐ and age‐matched wild‐type mice were used for confocal microscopy analysis.ResultsImmunohistochemical analysis of pathogen recognition receptors (PRRs) revealed an increase in levels of Toll‐like receptors 3 (TLR3) and 9 (TLR9), while those of NOD‐like receptor 3 (NLRP3) were decreased in NFE2L2/PGC1α ‐/‐ retina. Further analysis showed a significant increase in complement component C5a independent of component C3, tightly regulated by complement factor H (CFH). Interestingly, we found that thrombin, a serine protease enzyme, was involved in enhancing the terminal pathway producing C5a, independent of C3. We also detect an increase in primary acute phase C‐reactive protein (CRP) and receptor for advanced glycation end products (RAGE) in NFE2L2/PGC1α ‐/‐ retina.ConclusionsOur study reveals previously unknown thrombin mediated C3 independent activation of C5a in the dry AMD like retina of NRF2/PGC1α double knockout mice model.
p38/MAPK is required for protein aggregate formation and removal by autophagy upon proteasome dysfunction
Authors: Koskela Ali , Ruuth Johanna , Felszeghy Szabolcs , Kaarniranta Kai 
Published: January 2022

AbstractPurposeAge‐related macular degeneration (AMD) can be classified as an aggregation disease where cellular protein waste starts to accumulate in the retinal pigment epithelial (RPE) cells. Decline of autophagy and dysfunction of proteasomes have been identified as one the main reasons for the eventually detrimental protein aggregation. p38/mitogen‐activated protein kinases (MAPK) seem to play a regulatory role in autophagy. However, the function of p38/MAPK in proteasome dysfunction remains unveiled.MethodsARPE‐19 cells were treated with proteasome inhibitor MG‐132 (5 µM) to create a need for autophagosomal removal of protein waste. Simultaneously, commercial p38/MAPK inhibitor PD169316 (5 µM) were introduced to the cells to study it’s role in autophagy regulation. Autophagy flux was inhibited with Bafilomycin A1 (50 nM). Western blot for ubiquitin (marker of degradation‐destined proteins), p62 (protein aggregation marker) and LC3 (autophagosome marker) was performed. Similarly, immunocytochemistry of ubiquitin, p62 and LC3 to reveal their expression levels and cellular localization was conducted.ResultsProteasome inhibition resulted strong accumulation of ubiquitin‐tagged proteins, protein aggregates (p62) and autophagosomes (LC3‐II). p38/MAPK inhibition upon proteasome dysfunction blocked aggregate formation as p62 remained at basal level. Furthermore, despite the need for autophagosomal removal of protein waste due to proteasome inhibition, p38/MAPK inhibitor treated cells did not show increased autophagy. Immunocytochemistry revealed spread of ubiquitin‐ and p62‐tagged proteins throughout the cytosol upon p38/MAPK inhibition, autophagy‐related perinuclear accumulation of protein aggregates was not observed.Conclusionsp38/MAPK inhibition seemed to prevent protein aggregate formation upon proteasome inhibition and thereby autophagic degradation. Securing correct function of p38/MAPK may be a potential therapy target for aggregation diseases such as AMD.
The use of virtual three‐dimensional anatomy software in teaching ophthalmology
Authors: Scantling‐Birch Yarrow , Naveed Hasan , Okorie Michael , Rajak Saul 
Published: January 2022

AbstractPurposeMedical education is changing at both the undergraduate and postgraduate level. There are less hours dedicated to the basic sciences, especially anatomy. These changes threaten to marginalise ophthalmology education, which relies heavily on basic science knowledge for clinical application. Technology‐enhanced learning (TEL) can assist in the efficient knowledge transfer of basic ophthalmic knowledge. Our study scoped the literature and evaluated the use of a three‐dimensional (3D) virtual anatomy software to learn clinical and surgically relevant ocular anatomy at the undergraduate level.MethodsA literature search was conducted of the following databases: Embase, Medline and Cochrane Library. A national, cross‐sectional study was conducted via webinar teaching with 36 undergraduate medical students. A 60‐min lesson was used to teach ocular anatomy using a blended learning approach with novel virtual anatomy software (BioDigital, USA) and a didactic Powerpoint (Microsoft, USA) lecture. Anatomical knowledge gain was measured using validated multiple‐choice questions and a surgical video quiz. Student perceptions using questionnaires were measured before and after the online teaching session.ResultsThe literature showed a paucity of studies validating 3D virtual anatomy software in ophthalmology education. Software identified in the search included BioDigital, Essential Anatomy, Complete Anatomy and Visible Body. A significant improvement in mean knowledge gain was observed for both clinical and surgical quizzes after the use of BioDigital (17.9%, p < 0.0001). Significant confidence gain with respect to ocular anatomy was observed after the blended teaching session (40%, p < 0.0001). 100% of students enjoyed and agreed that 3D virtual anatomy software is an effective method of teaching ocular anatomy. 70% stated they will integrate digital anatomy software with their future ophthalmology learning, and 60% agreed the software could replace traditional teaching methods.ConclusionsThe declining importance given to ophthalmology teaching sets the tone for ophthalmic knowledge amongst graduating doctors and specialist trainees. TEL using 3D virtual anatomy software is an innovative, time‐efficient and student‐friendly teaching adjunct for ocular anatomy.
Diabetic db/db mouse presents intravitreal vessels protruding from the optic disc
Authors: Bonet Aina , Pampalona Judit , Segarra Irene , Ruberte Jesús 
Published: January 2022

AbstractPurposeIt is widely known that the main lesions that occur in diabetic retinopathy are oedema and neovascularization, but, still, there is not an accurate model that can reproduce both. In this study, we used a well‐established model of type 2 diabetes, the db/db mice, to assess the presence of retinal oedema and/or neovascularization.Methods12 weeks old db/+ and db/db mouse (n=8) were used in this study. We studied blood retinal barrier integrity by means of vascular leakage in fundus angiography by fluorescein injection and immunocytochemistry against albumin. Furthermore, we used transmitted electron microscopy to observe tight junction status. Finally, to visualize and assess retinal vascularization, eye fundus angiography and histology were performed.ResultsFluorescein and albumin remained inside the vessels proving the integrity of blood retinal barrier, in both controls and db/db retinas. However, retinal fundus angiography revealed that 50% of db/db mice presented intravitreal vessels protruding from the optic disc. Moreover, histological analysis showed a significant increment in the number of vessels in the db/db vitreous compared to db/+, in accordance with the angiography findings.ConclusionsRetinal fundus angiography and histological analysis showed the presence of intravitreal vessels in diabetic db/db mice, suggesting the existence of a current angiogenic process. However, apparently, twelve old week’s db/db mice did not show signs of retinal oedema.
Epithelial lip mucosa transplant re‐populates corneal anterior surface. An ex vivo study
Authors: Gerasimov Maxim , Ostrovskiy Dmitriy , Shatskikh Anna , Borzenok Sergey , Malyugin Boris 
Published: January 2022

AbstractPurposeTo establish a new ex vivo model and study the labial mucosal epithelium grafting as a potential approach for cornea re‐epithelization in patients with bilateral limbal stem cell deficiency.MethodsFour human donor corneal buttons were collected from the Eye tissue bank in corneal cold storage solution and left at 4°C for 32–52 days. Labial oral mucosa strips were dissected from four patients during fornix reconstruction after they signed an informed consent. The labial mucosa epithelial graft was prepared by dissecting substantia propria. The resulted graft was sutured near the limbus of the corneal button with running sutures (construct). Each construct was cultured under the standard conditions with the concave side upwards. After 21 days of culture, constructs were removed, washed and fixed. First half of the constructs were stained with hematoxylin and eosin (HE). 5 um crosscut cryo‐sections of the second half were stained with anti‐ keratin 13, 19, and p63, and paired secondary fluorescent antibodies. Nuclei were counterstained with Hoechst. Sections were accessed and photographed.ResultsAfter the culture, all constructs were integral with the attached epithelium graft and non‐loosened sutures. The graft became slightly flattened and brighter. HE staining demonstrated absence of the native cells in donor corneas. The labial mucosa grafts were located on the surface of the Bowman's membrane (BM) at the cornea periphery. Epithelial outgrowths were found to be transit from the labial mucosa graft to the BM. Cellular outgrows of the constructs had positive staining for keratin 13 and 19. Few cells were p63‐positive in nuclei.ConclusionsIn the study, a new ex vivo model composed of processed human donor cornea and live human donor labial mucosal epithelium graft was successfully established. The study confirmed the consistency of the new approach for corneal re‐epithelization by transplantation of an oral mucosal epithelial graft.
Endothelial cell viability after DMEK graft preparation
Authors: Miron Alina , Sajet Anita , Groeneveld‐van Beek Esther A , Kok Jet , Dedec Mehtap , de Jong Maloeke , Melles Gerrit R.J. , Oellerich Silke , van der Wees Jacqueline 
Published: January 2022

AbstractPurposeTo evaluate the effect of graft preparation and organ‐culture storage on endothelial cell density (ECD) and viability of Descemet membrane endothelial keratoplasty (DMEK) grafts.MethodsDMEK grafts (n = 27) were prepared at Amnitrans EyeBank Rotterdam from 27 corneas (15 donors) that were eligible for transplantation but could not be allocated due to the Covid‐19‐related cancellation of elective surgeries. Cell viability (by Calcein‐AM staining) and ECD of 5 grafts originally scheduled for transplantation, were evaluated on the originally planned surgery day, whereas 22 grafts from paired donor corneas were evaluated either directly post‐preparation or after 3–7 days of storage. ECD was analyzed by light microscopy (LM ECD) and Calcein‐AM staining (Calcein‐ECD)ResultsLight microscopy (LM) evaluation of all grafts showed an unremarkable endothelial cell monolayer directly after preparation. However, median Calcein‐ECD for the 5 grafts initially allocated for transplantation was 18% (range 9–73%) lower than median LM ECD. For the paired DMEK grafts, Calcein‐ECD determined by Calcein‐AM staining on the day of graft preparation and after 3–7 days of graft storage showed a median decrease of 1% and 2%, respectively. Median percentage of central graft area populated by viable cells after preparation and after 3‐7 days of graft storage was 88% and 92%, respectively.ConclusionsCell viability of most of the grafts will not be affected by preparation and storage. Endothelial cell damage may be observed for some grafts within hours after preparation with insignificant additional ECD changes during 3–7 days of graft storage. Implementing an additional post‐preparation step in the eye bank to evaluate cell density before graft release for transplantation may help to reduce postoperative DMEK complications
A multicentre randomised trial comparing the efficacy and safety of Remogen® Omega versus Cationorm® in the treatment of patients suffering from dry eye
Authors: Pisella Pierre‐Jean , Marsousi Niloufar , Doan Serge , Douat Julien , Girard Brigitte , Cochener Béatrice , Christophe Baudouin 
Published: January 2022

AbstractPurposeRemogen® Omega is the first omega‐3 containing drop indicated in the treatment of dry eye syndrome (DES). It contributes to the maintenance of the physicochemical balance of the tear film and facilitates eye recovery. The aim of this study was to compare the efficacy and safety of Remogen® Omega and Cationorm® in patients with moderate to severe DES.MethodsThis was a randomised controlled, masked‐observer multicentre trial. Patients used 4 drops of Remogen® Omega or Cationorm® per day for 3 months. The primary efficacy endpoint was the change from baseline at D28 in the score of ocular surface staining with fluorescein (OSSF) using Oxford scale. Other efficacy parameters included the score of ocular surface staining with lissamine green, Schirmer I test, tear film break‐up time, ocular surface disease index score, symptoms intensity and frequency, treatment comfort and overall efficacy at D28 and D84. Safety was evaluated based on intraocular pressure, best‐corrected visual acuity, general external examination, slit lamp examination and occurrence of adverse events.ResultsIn total, analysed intent‐to‐treat population consisted on 23 patients who were randomly assigned to Remogen® Omega (n = 10) or Cationorm® (n = 13) arms. Patients were in average 67.1 years old and 91.3% were women. Demographics were balanced between the two arms. Mean (standard deviation) reductions of ‐1.0 (1.1) and ‐0.7 (1.7) were observed for OSSF scores in Remogen® Omega (p = 0.01) and Cationorm® (p = 0.05) arms at D28 versus baseline, respectively. Similarly, improvements were observed in both treatment groups for other efficacy criteria. The only between‐arm difference detected was the duration of blur vision which was longer after administration of Remogen® Omega compared with Cationorm® at D28 and D84 (p < 0.01). Both treatments were considered as comfortable and satisfying by patients and investigators. No statistically significant between‐arm differences were observed for safety parameters and no safety issues were raised.ConclusionsObserved results suggest that both treatments have a beneficial effect on DES symptoms alleviation.
Ultrastructure of collagen fibrils, proteoglycans and lamellae of the human cornea treated with Iontophoresis ‐ UVA cross‐linking and hypotonic riboflavin solution
Authors: Akhtar Saeed , Almubrad Turki , Smedowski Adrian , Samivel Ramachandran , Almutleb Essam , Alkanaan Aljoharah , Khan Adnan , Masmali Ali , Mencucci Rita 
Published: January 2022

AbstractPurposeTo investigate the effect of iontophoresis‐UVA cross‐linking with hypotonic riboflavin solution on the ultrastructural changes in the lamellae, collagen fibrils (CF) and proteoglycans (PGs) in the central and peripheral stroma of the human corneal buttons.MethodsIontophoresis method was used for the trans‐epithelial application of hypotonic riboflavin in ex vivo corneal culture for 5 minutes. The corneas were irradiated using three methods; Group 1 (G1): an UV‐A irradiance of 3 mW/cm2 for 30 minutes; Group 2 (G2): an UV‐A irradiance of 10 mW/cm2 for 9 minutes; Group 3 (G3): without UV‐A irradiation. Three untreated corneas were used as a control group (G0). After CXL procedure, the corneas were processed for electron microscopy. CF diameter and PGs in each sample were analyzed using iTEM program.ResultsThe keratocyte organelles and stromal architecture in the peripheral cornea were better preserved than in the central cornea. The mean CF diameter in the peripheral cornea of the ‘G1 and G2’ was significantly higher compared to those in the central cornea. In G3, the central corneal CF diameter was significantly larger than the peripheral corneal CF diameter. There were also differences in the PGs area size between the central and peripheral cornea in all groups.ConclusionsRiboflavin + UVA application at 3mW2 /cm for 30minutes and 10mW2/cm for 9 minutes is a suitable method of CXL; however, the 10mW2/cm for 9 minutes combination seems to present stronger phototoxic effect and better prognosis for corneal healing. CXL treatment was more effective at the periphery than center.AcknowledgmentsThis research project was funded by National Plan for Science and Technology Program (Grant Number 15MED4229‐02), King Saud University.