bluuebunny/crossref_metadata_2025_split huggingface dataset:
In vitro and in vivo evaluation of genetically corrected RPE cells for autologous cell therapy of hereditary retinal dystrophies
Authors: Rodriguez‐Jimenez F. J. , Brymova A. , Mueller B. , Stieger K. , Jendelova P. , Ardan T. , Studenovská H. , Lytvynchuk L. , Straňák Z. , Ellederová Z. , Juhás J. , Motlík J. , Petrovski G. , Erceg Lukovic D. 
Published: January 2024

Purpose: The generation of genetically corrected RPE cells from patient‐derived human induced pluripotent stem cells (hiPSCs) may provide a source for autologous therapeutically‐relevant therapy for this rare retinal dystrophy as well to create the human cellular disease models. Recently we generated a faithful RPE model from patient bearing mutation in MERTK gene and genetically corrected these cells using CRISPR/Cas9 system. For further application in humans is necessary in vivo validation for survival.Methods: The RPE cells from the genetically corrected patient‐derived hiPSC were characterized using immunocytological methods for the differentiation and polarization markers, electronic microscopy, RT‐PCR, TEER measurement, ELISA test, Western Blot and phagocytic assay. The implantations of RPE‐scaffolds were carried out into the minipig eyes. Due to the xenogeneic transplantation of human cells into minipig eyes, tacrolimus immunosuppression therapy was used. At the 2‐week follow‐up from transplantation, non‐invasive OCT and fundus camera examinations of the implanted RPE‐scaffolds were performed. Consequently, euthanasia, histological and immunohistochemical investigations of the implanted retina were performed.Results: All generated RPE cells exhibit typical features of RPE cells. We demonstrated the reestablishment of the expression of full‐length MERTK protein as well as the reversion of lost phagocyte function of hiPSC‐RPE in vitro, which represents the first example of its kind in this field. Histological and immunohistochemical investigation of the RPE implants and adjacent retina showed structurally healthy RPE‐scaffold implant and neuroretinal cells above the implant.Conclusions: The generated hiPSC‐derived RPE in clinical grade conditions display the typical characteristics of mature RPE cells re‐establishing the function of phagocytosis in genetically corrected patient‐derived RPE cells in vitro. Functional in vivo studies in minipigs confirmed the RPE survival as well as survival of adjacent photoreceptors and thus can be employed as a potential treatment for patients.
The importance of having a relevant model system to investigate splicing modulation in the retina: The unforeseen cases of the intronic variants in RPE65 and IMPG2 genes
Authors: Domínguez Irene Vázquez 
Published: January 2024

Inherited retinal diseases (IRDs) are clinically and genetically heterogeneous disorders that can be caused by pathogenic variants in one of the more than 270 genes associated with IRDs. Among them, mutations in RPE65 can lead to Leber congenital amaurosis (LCA) whereas mutations in IMPG2 to vitelliform macular dystrophy (VMD).During my talk, I will provide evidence of the importance of employing the right cellular models to identify disease mechanism(s) and eventually evaluate suitable treatments. For this, I will highlight two intriguing examples of potential splicing defects studied by our group. Such defects are attractive for splicing modulation therapies using antisense oligonucleotides (AONs), a group of versatile molecules that can alter splicing by interacting with the target sequence(s) at pre‐mRNA level. Thus, characterizing the effect of a variant at the RNA level is instrumental to assess the applicability of AON‐based therapeutic intervention.We explored the effect of the homozygous variant c.11 + 5G>A in RPE65 and a heterozygous complex allele consisting of c.3023‐15T>A and c.3023G>A (p.(Gly1008Asp)) in IMPG2. In silico predictions suggested that these variants lead to a splicing defect. To assess the pathogenic effect of these variants, splice reporter assays using midigenes were conducted in HEK293T cells. Initial results pointed towards a 124‐nt exon elongation for the RPE65 variant and an in‐frame deletion of 48‐nt at the beginning of exon 15 for the complex allele in IMPG2.To validate these results, blood cells from the patients harbouring these defects were reprogrammed to induced pluripotent stem cells (iPSCs). Subsequently, iPSCs were differentiated to the retinal cells in which these genes are endogenously expressed. For RPE65, iPSCs were differentiated into retinal pigment epithelium (RPE) cells (>90 days differentiation) or, for IMPG2, into photoreceptor precursor cells (PPCs, 30 days differentiation). For the RPE65 variant, a cell specific effect was observed: while a clear exon elongation was observed in the midigene system, in RPE, the expression of the gene was highly reduced. In addition, no protein was detected and the patient‐derived cells showed differences compared to the control line. For IMPG2, the 48‐nt deletion was detected in a lower percentage in patient‐derived PPCs compared to the levels observed in the midigene system. However, aberrantly spliced IMPG2 transcripts were highly elevated in patient‐derived PPCs in comparison to control PPCs.Overall, our work shows the importance of having the right cellular context to determine the effect of genetic variants on splicing in the retina.
Corneo‐conjunctival sensory neuron: Types, transduction channels and codification of nerve impulses
Authors: Gallar Juana 
Published: January 2024

The cornea and the conjunctiva are innervated by the peripheral projections of a small number of primary sensory neurons whose somata are located at the ophthalmic region of the trigeminal ganglion. Most trigeminal neurons innervating the ocular surface are small and medium size, and contain neuropeptides such as SP and CGRP. Functionally, ocular surface neurons have been classified into five main neuronal types, based on their response to multiple modalities of stimuli (mechanical forces, heat, cold, H+, chemical substances, etc.). Polymodal nociceptors (activated by several modalities of stimulus, mechanical energy, heat, H+, and chemical irritants), mechanonociceptors (activated by mechanical energy), cold thermoreceptors (activated by temperature reductions and hyperosmolarity) are found in the cornea, the sclera and the conjunctiva. Besides, low‐threshold mechanoreceptors (activated by mechanical forces of low intensity) are found in the corneoscleral limbus and the lid border, and pruriceptors (activated by pruritogen substances) have been described in the tarsal conjunctiva, close to the lid margin.The ability to respond to the different modalities of stimulus lies in the types of transducing channels that each type of neuron expresses at its peripheral nerve endings. Sensory transduction involves the conversion of the physical or chemical stimuli acting on the nerve terminal into electrical energy, that is, into a local change of membrane potential that, if it is large enough, will generate action potentials that will travel along the axon to be processed by the CNS. So that, nerve terminals with TRPV1 channels are directly gated by heat, H+, and several chemical irritants such as capsaicin; nerve terminals with Piezo2 channels will be activated by mechanical forces; TRPM8 channels provide nerve terminals sensitivity to cold and osmolarity increases; et cetera.Ocular sensory neurons also express other ion channels that contribute to coding, that is to help the sensory neuron to establish a correspondence between the characteristics of the stimulus (intensity, temporal course, etc.) and specific parameters of the firing it induces. Voltage‐gated Ca2+, Na+ and K+ channels as well as cyclic nucleotide‐gated channels, among others, contribute to define the firing frequency and pattern to encode the traits of the stimulus. Thus, the ion channels present in the nerve terminals of each class of ocular sensory neuron provide the neurons the ability to detect and encode one or several modalities of stimulus, which constitutes the origin of the different sensations evoked on the ocular surface.(Supported by PID2020‐115934RB‐I00 from DOI: MCIN/AEI/10.13039/50110001103, and CIPROM/2021/48 from the Generalitat Valenciana, Spain)
Ruthenium‐ 106 brachytherapy in the treatment of central choroidal melanoma
Authors: Grajewski Luise , Lothar Krause 
Published: January 2024

Aims/Purpose: Choroidal melanoma is the most common ocular malignancy. Depending on the size of the tumour and its location in relation different treatment options are available. Brachytherapy with β‐emitting Ruthenium‐ 106 plaque is an effective and widely used treatment. The treatment of juxtapapillary and juxtafoveolar choroidal melanoma is challenging because it involves the macula and optic nerve. Studies have shown that central choroidal melanomas have a high recurrence rate.Methods: A total of 56 eyes of 56 patients with central choroidal melanoma were included in this retrospective study and treated with Ru‐106 brachytherapy between January 2011 and July 2020 in our institution. As central choroidal melanomas were defined all juxtafoveolar melanomas with a posterior edge touching an optic disc‐ fovea‐ radius around the fovea (about 5 mm). We measured visual acuity, recurrence, radiation related side effects and metastasis. The mean follow‐ up time was 66 (6–136) months.Results: Of the 56 eyes, 8 eyes (14%) showed a recurrence during the follow‐ up period. It was shown that the choroidal melanomas closer to the optic disc have a higher recurrence rate than the once close to the macula (69% vs. 95%). A radiation retinopathy was detected in 7 eyes (13%), a radiation opticopathy in 7 eyes (13%) and a radiation maculopathy in 10 eyes (17.9%). 6 eyes (11%) had to be enucleated because of recurrence or radiation complication.Conclusions: Ruthenium brachytherapy is an effective therapy for central choroidal melanoma with a rate of preserving the eye of 89%. The localization of the choroidal melanoma near the optic disc goes ahead with a higher rate of complications and recurrences.
Blunt dissection method using HiQ+ soft coagulation system for laparoscopic cholecystectomy in acute cholecystitis
Authors: Kawai Hironari , Omura Nobuo , Yamasawa Kaito , Tsukazaki Yuhei , Hirabayashi Tsuyoshi , Kawahara Hidejiro 
Published: January 2024

AbstractIntroductionDuring laparoscopic cholecystectomy for acute cholecystitis, it is often difficult to keep the surgical view dry because of inflammation‐related tissue fragility and susceptibility to bleeding. The resulting inadequate surgical view can lead to bile duct or vascular injury. Soft coagulation systems are used to achieve hemostasis during various surgeries; however, the usefulness of soft coagulation during laparoscopic cholecystectomy for acute cholecystitis is unclear. We here demonstrate the usefulness and feasibility of blunt dissection and soft coagulation during this procedure.Materials and Surgical TechniqueWe used blunt dissection and soft coagulation when performing laparoscopic cholecystectomy on two patients with acute cholecystitis. As with conventional laparoscopic cholecystectomy, four ports were inserted. After cutting the serosa by electrocautery, blunt dissection using soft coagulation was performed, exposing the inner subserosa. Maintaining this layer using blunt dissection with soft coagulation achieved a sufficiently clear view for safety. After resecting the cystic artery and duct, the gallbladder bed was also dissected by blunt dissection with soft coagulation. Blood loss was <20 mL in both patients.DiscussionBlunt dissection with soft coagulation may be a useful and feasible means of keeping the surgical view dry and minimizing blood loss during laparoscopic cholecystectomy for acute cholecystitis.
Unleashing digital superheroes: Unravelling the empathy factor in digital competence and online teacher autonomy support
Authors: Chen Anni , Li Wei , Fu Weidong 
Published: January 2024

AbstractConsiderable research has been dedicated to studying teachers' digital competence, yet limited insights have been gained regarding its impact on online teacher autonomy support. Based on the Technological Pedagogical Content Knowledge (TPACK) framework, this study utilized a multiple regression analysis model to explore how teachers' digital competence affects online teacher autonomy support. Drawing on data from 652 Chinese primary school teachers, the study revealed that teachers' digital competence positively influenced online teacher autonomy support via teachers' cognitive empathy. Teachers' emotional empathy strengthened the direct effect of cognitive empathy on online teacher autonomy support, and its indirect effect on teachers' digital competence on online teacher autonomy support. The findings inform TPACK framework development and practical considerations in online teaching, deepening understanding of the impact of digital competence on online teacher autonomy support.Practitioner notesWhat is already known about this topicPrevious research has recognized the importance of teachers' digital competence in supporting learners' autonomy and enhancing their online learning experiences.Existing studies have explored different dimensions of teachers' digital competence, including technological proficiency, pedagogical knowledge and content expertise.Studies have also examined the role of empathy in online teaching and learning, emphasizing its importance in fostering positive teacher–student relationships and promoting student engagement and motivation.What this paper addsThis study introduces digital competence as a novel and comprehensive assessment of technology integration in teaching, bridging an existing gap in the literature.This study recognizes the importance of socio‐emotional skills in online teaching by including teachers' empathic traits within the expanded TPACK framework.Teachers' cognitive empathy acts as a mediator and emotional empathy functions as a moderator in the relationship between digital competence and online teacher autonomy support, offering new insights into the underlying mechanisms.Implications for practice and/or policyTeacher educators should find ways to support teachers' digital competence in facilitating learners' autonomy in online learning environments effectively.Teacher educators should incorporate teacher empathy development into teachers' professional development programmes for online teaching, offering tailored interventions to differentiate and enhance both cognitive and emotional empathy.
Terrorism, counter‐terrorism, and voting: The case of Turkey
Authors: Yardimci M. Akif 
Published: January 2024

AbstractThis paper empirically tests the impact of terrorism and counter‐terrorism on voting by focusing on the two 2015 general elections in Turkey ‐ and the 2011 general election. This period offers a unique case because, after the first election, the ongoing peace process between the incumbent party (AKP), the political party associated with the perpetrators (pro‐Kurdish political party, HDP), and the imprisoned leader of the terrorist organization (PKK) was canceled. Instead, terror attacks recurred and curfews were implemented as counter‐terrorism measures. This enables the impact of curfews and terror attacks on electoral outcomes to be analysed in a difference‐in‐differences setting. Terror attacks are estimated to reduce the incumbent's vote share by 3.2% points, while increasing the vote share of the party associated with the perpetrators by 3.6% points. Curfews are estimated to cancel out the impact of terror attacks in attacked municipalities and decrease the incumbent's vote share by 4.7% points in non‐attacked municipalities.
The evolution of cnidarian stinging cells supports a Precambrian radiation of animal predators
Authors: Sierra Noémie C. , Gold David A. 
Published: January 2024

AbstractCnidarians—the phylum including sea anemones, corals, jellyfish, and hydroids—are one of the oldest groups of predatory animals. Nearly all cnidarians are carnivores that use stinging cells called cnidocytes to ensnare and/or envenom their prey. However, there is considerable diversity in cnidocyte form and function. Tracing the evolutionary history of cnidocytes may therefore provide a proxy for early animal feeding strategies. In this study, we generated a time‐calibrated molecular clock of cnidarians and performed ancestral state reconstruction on 12 cnidocyte types to test the hypothesis that the original cnidocyte was involved in prey capture. We conclude that the first cnidarians had only the simplest and least specialized cnidocyte type (the isorhiza) which was just as likely to be used for adhesion and/or defense as the capture of prey. A rapid diversification of specialized cnidocytes occurred through the Ediacaran (~654–574 million years ago), with major subgroups developing unique sets of cnidocytes to match their distinct feeding styles. These results are robust to changes in the molecular clock model, and are consistent with growing evidence for an Ediacaran diversification of animals. Our work also provides insight into the evolution of this complex cell type, suggesting that convergence of forms is rare, with the mastigophore being an interesting counterexample.
Comfort and wearability properties of custom‐made and boil‐and‐bite mouthguards among basketball players: A randomized parallel arm clinical trial
Authors: Doğan Özgür , Sönmez Hayriye , Doğan Suat Serhan Altıntepe 
Published: January 2024

AbstractBackground/AimThermoplastic mouthguards have been a reliable means of protection against impacts and shocks for a century. Although orofacial injuries are prevalent among basketball players, many athletes still avoid using mouthguards due to concerns regarding impaired breathing, speech interference, and nausea. This study aims to compare basketball players' subjective assessments of two different thicknesses of custom‐made mouthguards (MGs) with boil‐and‐bite MGs (B&B MG).Material and MethodsA total of 60 systematically healthy male professional basketball players, without active orthodontic treatment, were enrolled and randomly assigned to three groups: Group 1 (received B&B MGs; n = 20), Group 2 (received custom‐made MGs with a 3 mm thickness [ethyl vinyl asetat]; n = 20), and Group 3 (received custom‐made MGs with a 5 mm thickness; n = 20). To evaluate MG comfort and usability, 12 key factors, including comfort, fit, stability, tiredness, thirsting, oral dryness, nausea, speaking, breathing, drinking, ease of wear and removal, and inclination to chew, were assessed. The athletes were made to rate these factors using a 10 cm‐long Visual Analog Scale, measured at three‐time intervals, and the values for both inter‐group and intra‐group were compared.ResultsData from 48 basketball players were retrieved, and data analysis revealed that B&B MGs had the least favorable parameter values. Among the B&B MG group, stability was reported to be poorer during the initial measurement (p < .05). In the second measurement, B&B MGs showed significantly lower values for comfort, stability, tiredness, speaking, and inclination to chew (p < .05). During the third measurement, the B&B MG group exhibited significantly reduced values for comfort, breathing, drinking, and speaking (p < .05). In contrast, no statistically significant difference was observed between 3 and 5 mm thickness MGs in these measurements.ConclusionsCustom‐made MGs with different thicknesses consistently outperformed B&B MGs in all measurements, indicating the potential to tailor MG thickness based on sport, age, professional level of athlete, and presence of other protective equipment. While custom‐made mouthguards are considered the gold standard, dentists who provide B&B MGs can lead to cost savings while maintaining protection and encouraging athletes to use higher‐quality custom‐made mouthguards.
Effect of root immaturity and depth of intrusion on spontaneous re‐eruption and healing complications: A retrospective analysis
Authors: Singhal Ruchi , Negi Shefali , Namdev Ritu , Kakran Anisha 
Published: January 2024

AbstractBackground/AimsDental trauma resulting in tooth intrusion is a severe injury of permanent dentition, with a prevalence of 0.5%–1.9% among traumatic dental injuries. Currently, treatment guidelines for intruded permanent teeth depend on root development and the degree of intrusion. However, the categorization of tooth maturity as mature or immature simplifies a complex continuum of root development stages. This study aims to investigate the impact of various stages of root development on the success of spontaneous re‐eruption (SRE) of intruded teeth.Materials and MethodsThis retrospective study analysed data from 80 children (125 teeth) aged 6–12 years who experienced dental intrusion between 2018 and 2022. Root maturation was classified based on Cvek's classification, eight were categorized as stage 1, 29 as stage 2, 44 as stage 3, 25 as stage 4, and 19 as stage 5. Intruded teeth with immature roots (Cvek's class 1–4) underwent SRE, while mature teeth (Cvek's class 5) were treated with SRE, orthodontic repositioning, or surgical repositioning based on the degree of intrusion. Primary outcome measures were successful re‐eruption and secondary outcomes included observed complications during follow‐up.ResultsRegression analysis revealed that the patient's age, degree of root maturation, and degree of intrusion significantly affected spontaneous re‐eruption (p < .05). SRE was more successful in teeth with Cvek's stages 1 and 2 compared to stages 3 and 4. Complications were associated with the treatment method, degree of root immaturity, and degree of intrusion.ConclusionThe study demonstrates that as root maturation progresses, the likelihood of re‐eruption decreases, and the risk of pulp necrosis and infection increases. Therefore, teeth in later stages of immaturity (Cvek stage 4) should be repositioned orthodontically or surgically without waiting for spontaneous re‐eruption.
Electroencephalographic β‐band oscillations in the sensorimotor network reflect motor symptom severity in amyotrophic lateral sclerosis
Authors: Dukic Stefan , Fasano Antonio , Coffey Amina , Buxó Teresa , McMackin Roisin , Chipika Rangariroyashe , Heverin Mark , Bede Peter , Muthuraman Muthuraman , Lowery Madeleine , Carson Richard G. , Hardiman Orla , Nasseroleslami Bahman 
Published: January 2024

AbstractBackground and purposeResting‐state electroencephalography (EEG) holds promise for assessing brain networks in amyotrophic lateral sclerosis (ALS). We investigated whether neural β‐band oscillations in the sensorimotor network could serve as an objective quantitative measure of progressive motor impairment and functional disability in ALS patients.MethodsResting‐state EEG was recorded in 18 people with ALS and 38 age‐ and gender‐matched healthy controls. We estimated source‐localized β‐band spectral power in the sensorimotor cortex. Clinical evaluation included lower (LMN) and upper motor neuron scores, Amyotrophic Lateral Sclerosis Functional Rating Scale–Revised score, fine motor function (FMF) subscore, and progression rate. Correlations between clinical scores and β‐band power were analysed and corrected using a false discovery rate of q = 0.05.Resultsβ‐Band power was significantly lower in people with ALS than controls (p = 0.004), and correlated with LMN score (R = −0.65, p = 0.013), FMF subscore (R = −0.53, p = 0.036), and FMF progression rate (R = 0.52, p = 0.036).Conclusionsβ‐Band spectral power in the sensorimotor cortex reflects clinically evaluated motor impairment in ALS. This technology merits further investigation as a biomarker of progressive functional disability.