bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of the proteome changes in an in vitro hyperosmotic dry eye model employing human corneal epithelial cells
Authors: Lin Hao , Manicam Caroline , Pfeiffer Norbert , Grus Franz , Perumal Natarajan 
Published: January 2024

Aims/Purpose: The detrimental effects of tear hyperosmolarity on human corneal epithelial (HCE) cells is a primary cause of dry eye syndrome (DES). However, the molecular mechanisms underlying inflammatory processes and cellular dysfunction in HCE are largely unknown. This study characterized the proteome changes in an optimized hyperosmolarity‐induced DES model in vitro.Methods: An in vitro hyperosmolar (400–550 mOsm) DES model was established in a simian virus 40 (SV40)‐immortalized HCE cell line. Cells incubated in medium with physiological osmolarity (312 mOsm) were used as control. Cell viability, cell death and intracellular reactive oxygen species (ROS) levels were analysed with MTS, Annexin‐V‐FLUOS, crystal violet and DCFDA assays. Mass spectrometry‐based proteomics analyses characterized the proteome and elucidated the underlying molecular changes.Results: The HCE cell viability was significantly decreased in a concentration‐dependent manner with increasing hyperosmolarity compared to control at 48 hr (450 mOsm, −47%, p = 2.7 × 10−8; 500 mOsm, −71%, p = 1.1 × 10−4; 550 mOsm, −84%, p = 2.6 × 10−5). Notably, cell adherence was significantly decreased (p = 5.8 × 10−9), while levels of ROS (p = 3.6 × 10−2) and apoptosis (p = 3.6 × 10−2) were significantly increased at 450 mOsm compared to control. Proteomics analysis identified ~2000 proteins (FDR <1%) and, as many as 215 proteins were found to be significantly (p < 0.05) differentially abundant in the 450 vs. 312 mOsm. The effects of high hyperosmolarity (450 mOsm)‐induced damage resembling DES on the HCE cell were further demonstrated by the expression of proteins involved in the activation of inflammatory response (p = 5.5 × 10−3), apoptosis (p = 5.3 × 10−4) and generation of ROS (e.g., glutathione S‐transferase P) (p = 7.1 × 10−3). Moreover, many differentially expressed proteins were also implicated in the regulation of cell viability (p = 1.1 × 10−6) and organization of cytoplasm (e.g., cyclase associated actin cytoskeleton regulatory protein 1) (p = 2.4 × 10−2).Conclusions: In summary, an optimized hyperosmolarity‐induced DES cell culture model for proteomics analysis was established, and specific alterations in the proteome were characterized in this model. This in vitro model would be instrumental for testing therapeutic targets for DES in the future.
Allogeneic serum eye drops Sjogren syndrome‐related dry eye disease: Clinical evaluation of the ocular surface and in vivo confocal microscopy of the cornea
Authors: Forbice Eliana , Romano Vito , Semeraro Francesco , Pizzolante Teodoro , Passaro Maria Laura , Airaldi Matteo , Ancona Chiara , Bianchetti Andrea 
Published: January 2024

Aims/Purpose: To evaluate the symptomatic and clinical efficacy and in vivo confocal microscopy (IVCM) changes of the cornea after allogeneic serum eye drop therapy in patients with Sjögren's syndrome (SS)‐related Dry Eye Disease (DED).Methods: In this prospective clinical study 15 patients (30 eyes) with primary or secondary SS‐related DED were evaluated for Ocular Surface Disease Index (OSDI) questionnaire, ocular inflammation grade, Schirmer's test, tear film break‐up time (TBUT), corneal and conjunctival staining, and in vivo confocal microscopy (IVCM) analysis.Results: The average age of patients was 63.87 ± 9.18 years. The patients received treatment for a mean of 81.13 ± 56.84 days. The difference between before and after treatment with allogeneic serum eye drop therapy was statistically significant for OSDI parameters (p = 0.001), ocular inflammation (p = 0.004), TBUT (p = 0.007), Oxford Staining Scale (p = 0.007), Van Bijsterveld score (p = 0.006), basal epithelium density (p = 0.005), intermediate epithelium density (p = 0.041), Corneal Nerve Branch Density (CNBD) (p = 0.040), Beadings density (p = 0.047) and Fibre Tortuosity (p = 0.002).Conclusions: Allogeneic SEDs treatment in SS related DED patients, showed great results in terms of improvement in dry eye symptoms, clinical signs and IVCM changes. Further studies that aim at propagating and standardizing its production with clear international guidelines are needed.
Comparison between 100% autologous serum and 100% platelet‐rich plasma eye drops and their impact on the treatment efficacy of dry eye in primary Sjogren syndrome
Authors: Wróbel‐Dudzińska Dominika , Przekora‐Kuśmierz Agata , Kazimierczak Paulina , Kosior‐Jarecka Ewa , Ćwikińska‐Haszcz Agnieszka , Żarnowski Tomasz 
Published: January 2024

Purpose: The aim of the study was to compare the difference in the composition of 100% autologous serum (AS) and 100% platelet‐rich plasma (PRP) eye drops and assess their impact on the clinical out‐comes after the treatment of severe dry eye (DE) in primary Sjogren Syndrome patients (pSS).Methods: 22 patients with severe DE in pSS were treated with 100% AS (22 eyes) and 100% PRP (22 eyes) eye drops 5 times per day as monotherapy. The quantification of growth factors (GFs) as fibroblast growth factor (FGF), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), platelet‐derived growth factor (PDGF), nerve growth factor (NGF), transforming growth factor (TGF‐b), insulin‐like growth factor(IGF), fibronectin and substance P in hemoderivates was performed. The main outcomes: Ocular Surface Disease Index (OSDI), Best Corrected Visual Acuity (BCVA), the Schirmer test, tear break‐up time (TBUT), corneal and conjunctival staining according to the Oxford scale, conjunctival hyperaemia and Meibomian gland parameters. The results were compared at the baseline, 1 and 3 months after the treatment. The clinical results were correlated with the concentration of GFs.Results: Significant differences were observed in the concentration of FGF, EGF, fibronectin, VEGF, PDGF AB, NGF, PDGF, substance P, TGF‐β in PRP compared to AS. There was a statistically significant improvement in BCVA, the Schirmer test, TBUT, Meibomian gland parameters, and the reduction of the OSDI scores, Oxford staining and conjunctiva hyperaemia in each group. There were many correspondences between the level of GFs and the mean change of the clinical outcomes. No adverse events were reported.Conclusions: Despite the fact that blood derivatives differ in their composition, they seem to be effective, safe in the treatment of severe DE in pSS patients. The signs and symptoms of DE were reduced in both groups. The obtained clinical results and the composition of hemoderivates may indicate the superiority of PRP in relieving the signs and symptoms of DE in pSS patients.
Short‐term effect of rose bengal photodynamic therapy on collagen I, collagen V, nuclear factor kappa B, lysyl oxydase, transforming growth factor beta and interleukin 6 expression of human corneal fibroblasts, in vitro
Authors: Chai Ning , Stachon Tanja , Berger Tim , Li Zhen , Seitz Berthold , Langenbucher Achim , Szentmary Nora 
Published: January 2024

Aims/Purpose: To investigate collagen I, collagen V, nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB), lysyl oxidase (LOX), transforming growth factor β (TGF‐β) and interleukin‐6 (IL‐6) expression in healthy human corneal fibroblasts (HCFs) and keratoconus fibroblasts (KC‐HCFs), 24 h after rose bengal photodynamic therapy (RB‐PDT).Methods: HCFs were isolated from healthy human corneal donors (n = 5) and KC‐HCFs from elective penetrating keratoplasties (n = 5). Both cell cultures underwent RB‐PDT (0.001% rose bengal concentration, 0.17 J/cm2 fluence) and 24 h later collagen I, collagen V, NF‐κB, LOX, TGF‐β and IL‐6 mRNA and protein expression has been determined using qPCR and Western blot, compared to untreated cultures. IL‐6 concentration in the cell culture supernatant has been determined by ELISA.Results: TGF‐β mRNA expression was significantly lower (p = 0.02) and IL‐6 mRNA expression was significantly higher in HCFs 24 h after treatment (p = 0.01), than in controls. COL1A1, COL5A1 and TGF‐β mRNA expression was significantly lower (p = 0.048; p = 0.02 and p = 0.02) and IL‐6 mRNA expression was significantly higher (p = 0.02) in KC‐HCFs 24 h after RB‐PDT, than in untreated KC‐HCFs. TGF‐β protein expression in HCFs was significantly higher 24 h after treatment, than in controls (p = 0.03). IL‐6 protein concentration in the HCF and KC‐HCF culture supernatant was significantly higher 24 h after treatment, than in controls (p = 0.02; p = 0.03). Nevertheless, collagen I, collagen V protein, NF‐κB and LOX mRNA and protein expression did not differ significantly between any of the RB‐PDT treated and untreated groups.Conclusions: In conclusion, our study demonstrates that RB‐PDT reduces collagen I, collagen V and TGF‐β mRNA expression, while increasing IL‐6 mRNA and protein expression in KC‐HCFs. In HCFs, RB‐PDT increases TGF‐β and IL‐6 protein level after 24 h. These observations may provide a basis for future research in this field.
PAX6, FABP5 and keratocytes‐characteristic marker expression in human limbal fibroblasts and keratocytes of congenital aniridia and healthy subjects, in vitro
Authors: Li Zhen , Stachon Tanja , Chai Ning , Suiwal Shweta , Fries Fabian , Seitz Berthold , Amini Maryam , Shi Lei , Szentmary Nora 
Published: January 2024

Aims/Purpose: To investigate PAX6, FABP5 and keratocyte‐characteristic marker expression in human limbal fibroblasts and keratocytes (LFCs and LKCs) of congenital aniridia (AN) and healthy corneas, in vitro.Methods: Primary human limbal fibroblasts (LFCs/ AN‐LFCs) were isolated from aniridia (n = 8) and healthy corneas (n = 8) and were cultured in DMEM/F12 with 5% fetal calf serum. Limbal keratocytes (LKCs/ AN‐LKCs) were cultured in low‐glucose serum‐free cell culture medium (LGSF‐medium). Stem cell marker PAX6 and ABCG2, retinoic acid marker FABP5 and keratocyte‐characteristic marker collagen I (COL1A1), collagen III (COL3A1), collagen V (COL5A1), lumican (LUM), keratocan (KER), α‐smooth muscle actin (ACTA2), CD34, aldehyde dehydrogenase 3 family, member A1 (ALDH3A1) expression analysis has been performed by qPCR and Western blot.Results: PAX6 mRNA expression was significantly lower in AN‐LFCs and AN‐LKCs than in LFCs and LKCs (p = 0.04; p = 0.014). FABP5 mRNA expression was significantly higher in LKC and AN‐LKCs than in LFCs and AN‐LFCs (p = 0.036; p = 0.012) COL5A1 mRNA expression was significantly lower (p = 0.007), KER and ALDH3A1 mRNA expression was significantly higher (p = 0.013; p = 0.007) in AN‐LFCs than in LFCs. COL1A1 and COL5A1 mRNA expression was significantly higher in AN‐LKCs than in LKCs (p = 0.009; p = 0.001). Protein expression did not differ significantly between LFCs/ AN‐LFCs or LKCs/ AN‐LKCs for any of the analysed markers (p ≥ 0.05).Conclusions: Our study confirmed altered gene expression in aniridia limbal fibroblasts and keratocytes compared to healthy controls. These may play an important role in development and progression of aniridia associated keratopathy.
Correlative microscopy (CLEM) of tarsal conjunctival scraping: A new opportunity in the diagnosis of microbial keratitis
Authors: Troisi Mario , Del Prete Salvatore , Troisi Salvatore , Marasco Daniela , Costagliola Ciro 
Published: January 2024

Aims/Purpose: To evaluate the efficacy of correlated Scanning Electron Microscopy (SEM)/Optical Microscopy (OM) analysis of conjunctival scrapings in diagnosing infectious keratitis with prior negative culture tests or ineffective broad‐spectrum antibiotic therapy.Methods: 102 eyes of 87 patients (45M, 42F) affected by keratitis were evaluated by cytological examination of scrapings of the superior tarsal conjunctiva in OM (Nikon Eclipse 50i) and SEM (JEOL), after 72 h of wash‐out. Inclusion criteria: age > 18 years, corneal epithelial defects from suspected infection, ineffectiveness of previous broad‐spectrum antimicrobial therapy or negative conjunctival swab. Exclusion criteria: ocular surgery <3 months, immunosuppression, allergic keratitis, severe dry eye, ocular herpes, ulcer >3 mm, anterior chamber involvement. Identification of inflammatory infiltrate (OM) and microorganisms (SEM) was performed by two independent operators with the same devices and methodology. Weekly clinical control with new conjunctival scraping for OM and SEM after cycles of therapy were carried out.Results: Pathogenic microorganisms were detected in 94/102 eyes: Acanthamoeba (22), Candida (18), Mycoplasma (15), Mycobacteria (13), Chlamydia (12), Micrococci (11), Pseudomonas (6), Cocci (6), Aspergillus (5), Fusarium (2); in 14 cases 2 or more different forms coexisted; in 2 cases a subsequent PCR test revealed the presence of HSV‐1; 3 cases showed signs of severe dry eye; other 3 marked eosinophilia. Operator agreement: 99%. The specific treatment for the germs detected determined in all cases clinical and microbiological resolution in 14–49 days (mean 27.6 ± 2.4).Conclusions: CLEM put in evidence the correlation between phlogistic cells and pathogens, allowing the diagnosis and the treatment of different forms of infectious keratitis and other pathological conditions. The interpretation and repeatability, however, require appropriate training and dedicated equipment.
Segmentation of the fibrillar layer in advanced fuchs endothelial corneal dystrophy: Inter‐oberserver‐reliability and intra‐observer‐repeatability analysis of corneal densitometry images
Authors: Howaldt Antonia , Mestanoglu Mert , Musial Gwen , Jonas Rahul , Cursiefen Claus , Bachmann Björn , Matthaei Mario 
Published: January 2024

Aims: Our previous studies state that a central subendothelial collagen‐rich fibrillar layer (FL) is present in about 80% of advanced Fuchs endothelial corneal dystrophy (FECD) corneas which correlates with decreased corneal endothelial cell density and focal oedema formation. In this study, we investigated the previously unknown inter‐observer reliability and intra‐observer reproducibility of FL segmentation on Scheimpflug corneal densitometry images in patients with advanced FECD.Methods: Preoperative Scheimpflug images of patients receiving DMEK or triple DMEK for advanced FECD were retrospectively analysed. FL‐negative cases were excluded. Images were manually segmented with the open‐source software MITK by two experienced graders. A MATLAB‐based code calculated the region of interest (ROI) area, maximum calliper length, maximum horizontal and vertical dimension, and Dice similarity coefficient for FL‐areas. The intraclass correlation coefficient (ICC) was calculated by IBM SPSS (two‐way mixed, average measures). The FL localization on corneal densitometry were visualized by spatial heatmaps by MATLAB.Results: Among 308 eyes, 248 were FL‐positive (80.5%) and 60 FL‐negative (19.5%). The average Dice similarity coefficient of FL areas segmented by two graders was calculated at 0.85. Calculations on the segmentations of Grader 1 vs. Grader 2 include: ROI 7.91mm2 ± 4.8 vs. 9.29 mm2 ± 5.0, Calliper length 4.07 mm ± 1.1 vs. 4.21 mm ± 1.0, maximum horizontal dimension 3.85 mm ± 1.1 vs. 3.95 mm ± 1.0, maximum vertical dimension 3.17 mm ± 1.0 vs. 3.3 mm ± 1.0. The ICC was calculated at 0.938 vs. 0.957 vs. 0.930 for ROI (0.955 vs. 0.960 vs. 0.944 calliper length, 0.958 vs. 0.959 vs. 0.946 maximum horizontal dimension, 0.952 vs. 0.941 vs. 0.945 maximum vertical dimension) and 0.85 vs. 0.86 vs. 0.89 for the Dice similarity coefficient (inter‐observer vs. intra‐observer Grader 1 vs. intra‐observer Grader 2). Spatial heatmap analysis demonstrated primarily inferotemporal localization of increased densitometry areas in FL‐positive eyes.Conclusions: FL segmentation on corneal densitometry images exhibit high inter‐observer reliability and intra‐observer reproducibility scores. We provide evidence for corneal densitometry images as a valid tool to detect FL in advanced FECD and confirm the inferotemporal corneal projection of FL.
Neuroinflammatory findings with corneal COnfocal microscopy in long COVID‐19 patients, 2 years after acute SARS‐CoV‐2 infection
Authors: Suarez Pilar Canadas , Gonzalez‐Vides Leonela , García‐Velasco Marta Alberquilla , Verdejo José Luis Hernández 
Published: January 2024

Purpose: To describe corneal confocal microscopy findings in patients with long COVID‐19 with persistent symptoms over 20 months after SARS‐CoV‐2 infection.Methods: Cross‐sectional comparative transversal study that included a total of 53 patients, 33 patients with Long COVID‐19 and 22 controls matched in age and gender. Long COVID‐19 diagnosis was established according to the World Health Organization criteria Corneal confocal microscopy Heidelberg Retina Tomograph II (Heidelberg Engineering, Heidelberg, Germany) was performed to evaluate sub‐basal nerve plexus morphology (corneal nerve fibre density, nerve fibre length, nerve branch density, nerve fibre total branch density, nerve fibre area, and nerve fibre width). Dendritic cells density and area, along with microneuromas and other morphological changes of the nerve fibres were recorded.Results: Long COVID‐19 patients presented reduced corneal nerve density and branch density as well as shorter corneal nerves compared to the control group. Additionally, Long COVID‐19 patients showed an increased density of Dendritics Cells with also greater area than that found in the control group of patients without systemic diseases. Microneuromas were detected in 24.2% of Long COVID‐19 patients.Conclusions: Long COVID‐19 patients exhibited altered corneal nerve parameters and increased DC density over 20 months after acute SARS‐CoV‐2 infection. These findings are consistent with a neuroinflammatory condition hypothesized to be present in patients with long COVID‐19, highlighting the potential role of corneal confocal microscopy as a promising noninvasive technique for the study of patients with long COVID‐19.
“Mucus fishing syndrome”: Presentation of this rare clinical entity along with its therapeutic options
Authors: Mancini Alessandra , Scorcia Vincenzo , Giannaccare Giuseppe 
Published: January 2024

Aims/Purpose: The “Mucus fishing syndrome” (MFS) is a rare chronic inflammatory condition involving the ocular surface characterized by the continuous formation, of mucus filaments. The clinical presentation is variable and various underlying ocular surface pathologies including dry eye, blepharitis and allergic conjunctivitis can be associated. Chronic irritation, conjunctival hyperaemia and mucus production are present. On physical examination, patients may present an iatrogenic damage of the conjunctival epithelium due to the mechanical removal of mucus and a vicious circle is usually established which causes further conjunctival irritation and more conspicuous mucous secretion. The treatment involves the elimination of abnormal mucus production and the education of the patient to avoid contact with the ocular surface during the manoeuvres. We present a case of MFS in a young woman affected by meibomian gland dysfunction.Methods: The diagnosis was reached due to the presence of persistent mucous discharge and the characteristic rose bengal staining pattern. Initial supportive treatment (Tear substitutes and continuous irrigation of the ocular surface) prescribed elsewhere did not provide clear clinical benefits. Therefore, upon presentation a new approach was pursued adding to the ongoing treatment hydrocortisone eye drop 3 times a day, and a mucolytic agent (5% acetylcysteine eye drop) 4 times a day, together with accurate eyelid hygiene (morning and evening). Patient was also informed to avoid mechanical removal of the mucous.Results: One month after therapy the presence of mucus was reduced and the symptoms improved.Conclusions: Mucus fishing syndrome is a rare but challenging condition affecting the ocular surface. The objective of the therapy should address the pharmacological lysis of the mucous produced along with the instruction of patient not to compulsively remove the filaments in order to avoid further stress and accelerate healing.
Meibomian gland dysfunction in people who have been in the refuge for a long time during the hostilities in Ukraine
Authors: liudmyla Liudmyla , Bezditko Pavel , Boieva Yuliia 
Published: January 2024

Aims/Purpose: To study the indicators of the eye surface in people who have been in the refuge for a long time during the hostilities in Ukraine.Methods: We recruited 58 patients (main group) aged 32–75 years, who have been in the basement or underground from 3 to 5 month during the hostilities in Ukraine. The control group – 42 patients, aged – 32–72 years, who have been in their houses or flats. Each patients underwent additional examination: the Schirmer's test, the Norn's test, determination of the ocular protection index (OPI), contact meibography (Pult et al., 2012), compression test for evaluation of excretory capacity (Korb, 2005) and quality of the secretion (The International Workshop on Meibomian Gland Dysfunction, 2011) of meibomian gland, and Ocular Surface Disease Index Questionnaire (OSDI). The International Workshop's Classification of Meibomian Gland Dysfunction, 2011 was used to evaluate the severity of MGD.Results: In 75.8% patients of the main group Schirmer test was 4.8 ± 0.7 mm, Norn test was 4.1 ± 0.8 s, OPI test was 0.54 ± 0.8, test for evaluation of excretory capacity and quality of the secretion of meibomian glands was 2 stage, the mean of OSDI was 56.6 ± 3.4 (p < 0.05). In 71.4% patients of the control group Schirmer test was 14.1 ± 1.6 mm, Norn test was 9.7 ± 0.8 s, OPI test was 0.69 ± 0.1, test for evaluation of excretory capacity and quality of the secretion of meibomian glands was 1 stage, the mean of OSDI was 11.2 ± 1.7 (p < 0.05).Conclusions: Our findings suggest that 75.8% of all patients who have been in the refuge from 3 to 5 month, have significant changes in the indicators of the eye surface, which corresponds to Meibomian Gland Dysfunction stage 3 and 4 (p < 0.05). 71.4% patients of the control group who have been in their houses or flats have changes in the indicators of the eye surface, which corresponds to Meibomian Gland Dysfunction stage 1 and 2 (p < 0.05). Long stays in the refuge caused significant changes in the surface of the eye, which required treatment for 75.8% of all patients (p < 0.001, respectively).