A bio-orthogonal linear ubiquitin probe identifies STAT3 as a direct substrate of OTULIN in glioblastoma.

While linear ubiquitin plays critical roles in multiple cell signaling pathways, few substrates have been identified. Global profiling of linear ubiquitin substrates represents a significant challenge because of the low endogenous level of linear ubiquitination and the background interference arising from highly abundant ubiquitin linkages (e.g. K48- and K63-) and from the non-specific attachment of interfering proteins to the linear polyubiquitin chain. We developed a bio-orthogonal linear ubiquitin probe by site-specific encoding of a norbornene amino acid on ubiquitin (NAEK-Ub). This probe facilitates covalent labeling of linear ubiquitin substrates in live cells and enables selective enrichment and identification of linear ubiquitin-modified proteins. Given the fact that the frequent overexpression of the linear linkage-specific deubiquitinase OTULIN correlates with poor prognosis in glioblastoma, we demonstrated the feasibility of the NAEK-Ub strategy by identifying and validating substrates of linear ubiquitination in patient-derived glioblastoma stem-like cells (GSCs). We identified STAT3 as a bona fide substrate of linear ubiquitin, and showed that linear ubiquitination negatively regulates STAT3 activity by recruitment of the phosphatase TC-PTP to STAT3. Furthermore, we demonstrated that preferential expression of OTULIN in GSCs restricts linear ubiquitination on STAT3 and drives persistent STAT3 signaling, and thereby maintains the stemness and self-renewal of GSCs.

Mutation analysis of TGFBI and KRT12 in a case of concomitant keratoconus and granular corneal dystrophy.

PURPOSE: This study is to summarize the concurrent keratoconus (KC) and granular corneal dystrophy (GCD) phenotype and identify the underlying genetic cause in a 23-year-old male patient. METHODS: A detailed family history and clinical data from the patient and his parents were collected by ophthalmologic examination. The candidate genes were captured and sequenced by targeted next-generation sequencing, and the results were confirmed by Sanger sequencing. RESULTS: The proband was clinically diagnosed as a case of concurrent KC and GCD, which is a very rare presentation. His father and grandmother were diagnosed as GCD in both eyes. There was no character of KC in his father's and grandmother's eyes. A heterozygous TGFBI mutation in exon 4 (c.370G > A) was identified in the proband, which was predicted to generate a missense mutation (p.R124H). The mutation also existed in his father and grandmother. A heterozygous KRT12 mutation in exon 8 (c.1456-1457ins GTA) was identified in the proband, which was predicted to generate an insert mutation and created a premature termination codon. The mutation did not exist in his father and grandmother. The two mutations did not exist in his mother and 200 unrelated normal controls. CONCLUSIONS: KC can co-exist with GCD. The missense mutation (c.370G > A) in the TGFBI gene and insert mutation (c.1456-1457ins GAT) in the KRT12 gene were identified in a 23-year-old male patient with concurrent KC and GCD.

[Objective optical quality after orthokeratology].

OBJECTIVE: To determine the changes of objective optical quality after orthokeratology. METHODS: Prospective, self-controlled study. Optical quality, including objective scatter index (OSI), modulation transfer function (MTF) cutoff frequency, contrast visual acuity (CVA) and objective tear film quality, was assessed with the OQAS II instrument before and 1 week, 1 month, and 3 months after orthokeratology. Parameters of corneal astigmatism and e value from topography were also assessed. The data was analyzed using one-way ANOVA. RESULTS: Cylinder refraction was (-0.39 ± 0.39)D, (-0.65 ± 0.57)D, (-0.79 ± 0.56)D, and (-0.81 ± 0.53) D, respectively, at the four timepoints, and there was a statistically significant difference between before and after therapy (P < 0.05). Uncorrected visual acuity was 3.97 ± 0.51, 4.95 ± 0.12, 5.00 ± 0.06, and 5.00 ± 0.05, respectively. The e value was 0.37 ± 0.24, -0.23 ± 0.42, -0.29 ± 0.39, and -0.30 ± 0.37, respectively. The negative values after therapy were significantly different from the value before therapy (P < 0.01). The corneal astigmatism in 3 mm diameter was (1.27 ± 0.75) D, (1.06 ± 0.69) D, (1.11 ± 0.68) D , and (1.15 ± 0.67) D, respectively. There was no significant difference in astigmatism between before and after therapy (P ≥ 0.05). After therapy, the OSI increased from 0.59 ± 0.47 before therapy to 0.99 ± 0.66 at 1 week, 1.47 ± 1.17 at 1 month, and 1.49 ± 1.03 at 3 months. The difference between pre- and post-therapy OSI values was statistically significant (P < 0.05). The MTF cutoff frequency decreased from (36.6 ± 12.0) cpd before therapy to (34.2 ± 8.9) cpd at 1 week, (27.9 ± 12.6) cpd at 1 month, and (27.7 ± 11.6) cpd at 3 months after therapy. The difference in MTF cutoff frequency between before and 1 month after therapy was statistically significant (P < 0.05). The 100% CVA, 20% CVA, and 9% CVA all declined after therapy, but there was no statistically significant difference between before and after therapy. The OSI of tear film quality was 1.27 ± 1.12, 2.09 ± 1.67, 2.07 ± 1.12, and 2.10 ± 1.33, respectively, and the difference was statistically significant between before and after therapy (P < 0.05). CONCLUSIONS: After 3 months of orthokeratology, the corneal central astigmatism remained, and the e value became negative, while OSI increased, and MTF cutoff frequency and tear film quality declined. The alteration was more and more obvious along with the therapy within 1 month, and was not significant within the following month. After short-term corneal reshaping, the ocular optical quality declined in some degree, but the loss was acceptable.

Efficacy of orthokeratology lens with the modified small treatment zone on myopia progression and visual quality: a randomized clinical trial.

BACKGROUND: To evaluate the long-term effectiveness of orthokeratology (ortho-K) lenses with small treatment zone (STZ) or conventional treatment zone (CTZ) in controlling axial elongation in children with myopia as well as the impact on visual quality. We also sought to determine the effect of retinal visual signal quality on axial elongation. METHODS: This is a prospective randomized controlled study. A total of 140 participants (age ranging from 8 to 12 years) were randomly assigned to wear either STZ or CTZ ortho-K lenses. STZ ortho-K lenses design was achieved by changing the depth of reverse zone and the sagitta height of the optical zone. Using the IOL-Master 500, axial length (AL) was measured at baseline and after 6, 12 and 18 months of ortho-K treatment. Spherical aberration (SA) and corneal topographic parameters were obtained by the Pentacam anterior segment analyzer at baseline and the 1-month follow-up visit, and optical qualities were assessed by optical quality analysis system-II (OQAS-II) at baseline and after 1 month of lens wearing. Optical quality parameters mainly included the modulation transfer function (MTF) cutoff, Strehl ratio (SR), objective scattering index (OSI), and predicted visual acuity (PVA). RESULTS: A total of 131 participants completed the study, including 68 in the STZ group and 63 in the CTZ group. The STZ group had significantly reduced AL elongation compared to the CTZ group after treatment (12 months: 0.07 ± 0.11 mm vs. 0.14 ± 0.12 mm, P = 0.002; 18 months: 0.17 ± 0.15 mm vs. 0.26 ± 0.16 mm, P = 0.002). The topography in the STZ group showed a smaller treatment zone (TZ) diameter (2.50 ± 0.23 mm vs. 2.77 ± 0.18 mm, P < 0.001), a wider defocus ring width (2.45 ± 0.28 mm vs. 2.30 ± 0.30 mm, P = 0.006), and larger values of total amount of defocus (119.38 ± 63.71 D·mm2 vs. 91.40 ± 40.83 D·mm2, P = 0.003) and total SA (0.37 ± 0.25 µm vs. 0.25 ± 0.29 µm, P = 0.015), compared with the CTZ group. Objective visual quality decreased in both groups (P < 0.001). This was evidenced by a greater decrease in MTF cutoff (- 14.24 ± 10.48 vs. - 10.74 ± 9.46, P = 0.047) and SR values (- 0.09 ± 0.07 vs. - 0.06 ± 0.07, P = 0.026), and an increase in OSI value (0.84 ± 0.72 vs. 0.58 ± 0.53, P = 0.019). PVA9% decreased significantly in the STZ group but not the CTZ group. A statistically significant negative correlation was found between the changes in total SA and MTF cutoff values (r = - 0.202, P = 0.025). AL changes were associated with sex, change of MTF cutoff value, increment of total SA and TZ area. CONCLUSIONS: Compared with CTZ ortho-K lenses, STZ ortho-K lenses significantly inhibited axial elongation in children with myopia while moderately reducing their objective visual quality. Axial elongation was affected by retinal visual quality, and it may be a possible mechanism for ortho-K slowing myopia progression. Trial registration This trial is registered at Chinese Clinical Trial Registry on November 5, 2019 with trial registration number: ChiCTR1900027218. https://www.chictr.org.cn/showproj.html?proj=45380.

Visual rehabilitation using rigid gas permeable contact lenses after femtosecond laser-assisted minimally invasive lamellar keratoplasty in patients with keratoconus.

This study aims to observe the clinical efficacy and safety of rigid gas permeable corneal contact lenses (RGP-CLs) wearing after femtosecond laser-assisted minimally invasive lamellar keratoplasty (FL-MILK) in progressive and advanced keratoconus eyes. Twenty-five patients (27 eyes) fitted with RGP-CLs after FL-MILK were enrolled, and 22 grading-matched keratoconus patients (23 eyes) as a control group. Corneal morphological data, diopter, best corrected vision, corneal endothelium, non-invasive tear film rupture time (NIBUT), corneal perception and comfort questionnaire were analysed before and after wearing RGP-CLs. In the FL-MILK group, the flat K, steep K and Kmax of the corneal anterior surface were decreased by 3.05D, 3.48D and 7.17D respectively after surgery (P = 0.011, 0.004 and 0.007). The central corneal thickness increased by 175.29 µm (P < 0.001). The basic curve of RGP-CLs after surgery was about 0.23 mm flatter than that before surgery (P = 0.013). There was no statistical difference in the best corrected vision and the comfort of wearing RGP-CLs before and after FL-MILK (P = 0.923, 0.391). Compared with the control group, there was no significant difference in vision improvement by RGP-CLs (P = 0.669). During the follow-up, the comfort of wearing RGP-CLs in the two groups was good, and no obvious adverse events were observed. This study showed MILK significantly increases corneal thickness and flattens corneal curvature, achieving good RGP-CLs corrected vision.

Parental Corneal Tomographic and Biomechanical Characteristics of Patients With Keratoconus.

PURPOSE: To investigate the hereditability of corneal tomographic and biomechanical parameters in keratoconus (KC). DESIGN: Prospective cohort study. METHODS: This study was conducted at Qingdao Eye Hospital of Shandong First Medical University in Qingdao, China. Forty-four patients with KC and their biological parents (n = 88) were recruited as the study group. The control group consisted of 84 healthy adults with matched age and gender. Both eyes of each participant underwent clinical examinations, and 1 eye was selected for statistical analysis. Exclusion criteria were as follows: individuals with glaucoma, ocular surgery, systemic diseases known to affect the eyes, or poor cooperation during examination. Subjects were asked to discontinue soft contact lens (CL) wear for 2 weeks and rigid gas permeable CL wear for 4 weeks before ocular examination. All participants underwent a comprehensive assessment including Pentacam Scheimpflug tomography, Corvis ST, visual acuity, refraction examination, axial length, and slitlamp examination for both eyes. Individuals presenting with KC manifestations in at least 1 eye were classified as having KC. A total of 9 Pentacam indices including keratometry in the flat/steep meridian (K1/K2), maximal keratometry (Kmax), thinnest point pachymetry (TP), and maximum/average Ambrósio relational thickness (ARTmax/ARTave), anterior and posterior surfaces elevation of the cornea (Ef/Eb) and total deviation value (Final D), and 21 biomechanical indices were collected. Associations of these factors with KC were evaluated using multiple comparison and binary logistics regression analyses. RESULTS: Two parents (2.27%) from 2 different families were diagnosed with KC. Parents of patients with KC had thinner corneas with altered corneal biomechanical parameters compared with healthy controls (P < .05). The combined tomographic and biomechanical index demonstrated the highest discriminatory power (area under the receiver operating characteristic curve 0.785) and strong specificity (84.5%). Parental corneal tomographic and biomechanical index, Corvis biomechanical index, and TP were identified as the major influential factors for KC in their offspring by logistic regression analysis, with a 73.3% accuracy in identifying offspring with KC. CONCLUSIONS: Parental corneal tomographic and biomechanical properties of patients with KC suggest a possible predisposition to KC. A combination of tomography and corneal biomechanics can be helpful in predicting the incidence rate of KC in the offspring of patients with subclinical KC.

Modular Pathway Compartmentalization for Agroclavine Overproduction in Saccharomyces cerevisiae.

Agroclavine, which has anti-depressant activity and anti-Alzheimer effects, is the raw material used to synthesize ergo-based drugs. Although the production of agroclavine from Saccharomyces cerevisiae is possible, its yield is exceptionally low. The current study proposes a modular compartmentalization strategy for identifying and modifying the bottleneck step in agroclavine overproduction. The agroclavine synthetic pathway was reconstituted in yeast, and the best combination of Claviceps fusiformis EasA with Claviceps purpurea EasD/EasG was identified. According to the data on the expression and subcellular localization of agroclavine pathway proteins, the whole pathway was divided into two modules by chanoclavine-I. Separate enzyme distribution within the downstream module and low expression of DmaW and EasE in the upstream module were identified as the bottleneck steps in the pathway. The pathway efficiency was enhanced 2.06-fold when the downstream module was entirely anchored to the endoplasmic reticulum compartment. Increasing NADPH supply by overexpressing POS5 further improved the agroclavine yield by 27.4%. Altering the intracellular localization of DmaW from the peroxisome to the endoplasmic reticulum (ER) not only improved protein expression but also accelerated the accumulation of agroclavine by 59.9%. Integration of all modified modules into the host chromosome resulted in an improved yield of agroclavine at 101.6 mg/L with flask fermentation (a 241-fold improvement over the initial strain) and ultimately produced 152.8 mg/L of agroclavine on fed-batch fermentation. The current study unlocked the potential of S. cerevisiae in the advanced biosynthesis of ergot alkaloids. It also provides a promising strategy to reconstitute compartmentalized pathways.

Norepinephrine as an Enhancer Promoting Corneal Penetration of Riboflavin for Transepithelial Corneal Crosslinking.

Purpose: Previously, we found norepinephrine (NE) could affect the corneal epithelial integrity, herein we investigated the feasibility and safety of NE serving as a chemical enhancer to promote corneal penetration of riboflavin during transepithelial corneal crosslinking (CXL). Methods: The dosage of NE that could promote riboflavin diffusion through the healthy epithelial barrier without inducing epithelial damage in C57BL/6 mice was determined. The safety of NE treatment was confirmed by morphological and histological examinations of the whole cornea. The efficacy of NE in promoting riboflavin penetration was verified by slit lamp and scanning electron microscope (SEM), and corneal biomechanical measurement after CXL. To better fit the clinical scenario, increased NE dosage and shortened riboflavin infiltration time were further evaluated. Results: The lowest dosage of NE (1 mg/mL) that facilitated transepithelial riboflavin permeation was 2 µL. No visible corneal structure alteration was observed after NE treatment. SEM indicated dissociation of intercellular junctions among corneal epithelial cells. Homogenous distribution of riboflavin throughout corneal stroma was observed. NE-treated corneas reached comparable biomechanical properties after CXL, including stress-relaxation curve and elastic modulus, with corneas treated with the commercially available transepithelial drug Peschke TE. To better fit the clinical scenario, increasing NE up to 5.5 µL helped riboflavin infiltrate the corneal stroma within 30 minutes. After CXL with 9 mW/cm2 ultraviolet-A (UVA) for 2.5 minutes, the cornea showed significantly enhanced corneal biomechanical properties with undisturbed corneal endothelium. Conclusions: NE serves as an effective enhancer in increasing riboflavin diffusion with limited impairment on corneal epithelium and has great potential for clinical application. Translation Relevance: NE serves as an effective enhancer for riboflavin penetration and clinical transepithelial CXL.

Norepinephrine as the Intrinsic Contributor to Contact Lens-Induced Pseudomonas aeruginosa Keratitis.

Purpose: Contact lens wear (CLW) is one of the leading risk factors for Pseudomonas aeruginosa keratitis (PAK). However, the intrinsic factors that contribute to the high susceptibility to keratitis during CLW remain to be elucidated. CLW over an extended period can elevate corneal norepinephrine (NE) concentration. In this study, we investigated the role of NE in promoting PAK. Methods: We constructed an injury-induced PAK model and a CLW-induced PAK model to confirm the impact of NE during corneal infection. Pharmacological blockage of NE and gene knockdown mouse were used to investigate the downstream effector of NE. RNA sequencing was performed to explore the cellular alterations during NE treatment. Non-parametric Mann-Whitney U test or Kruskal-Wallis test were used to ascertain the significance (P < 0.05). Results: Supplementation of NE led to PAK even without artificial corneal injury during CLW. The effect was mediated by the ß2-adrenergic receptor (ß2-AR) in the corneal epithelium. The ß2-AR blockage by the NE antagonist ICI118,551 (ICI) or by deleting of its encoding gene Adrb2 significantly alleviated infection during CLW. Conversely, ß2-AR activation compromised the integrity of the epithelium and significantly increased the cortical plaque marker ezrin. Transcriptome analysis identified that the protective effect of ICI on the keratitis was mediated by dual-specificity phosphatases. Suramin, a Dusp5 antagonist, abrogated the protective effect of ICI. Conclusions: These data reveal a new mechanism by which NE acts as an intrinsic factor that promotes CLW-induced PAK and provide novel therapeutic targets for treating keratitis by targeting NE-ß2-AR.

[Comparison of Pentacam and OrbscanII systems for the diagnosis of keratoconus suspects].

OBJECTIVE: To compare the sensitivity of Pentacam and OrbscanII and to analyze the morphology characteristic of topography in normal eyes and different stages of keratoconus, to provide the basis for early screening of keratoconus suspects. METHODS: One hundred and ninety six normal eyes, 50 eyes with keratoconus suspect and 73 eyes with clinical keratoconus were enrolled. The changes of corneal anterior/posterior curvature, best fitting sphere (BFS). Inferior-Superior value (I-S) (3 mm and 5 mm), classification of elevation maps and corneal thickness were measured. RESULTS: There were significant differences including posterior curvature, anterior/posterior BFS, elevation value and corneal thickness in normal eyes between the data measured by these two different instruments, values obtained by Pentacam were less than those by Orbscan. Concerning the elevation maps pattern, the progressively decreasing pattern was dominant in Pentacam with normal eyes and keratoconus suspects, with the progressively increasing pattern accounting for 14.4% (28/196) and 20.0% (10/50) respectively in anterior surface, and 2.0% (41/96) and 26.0% (13/50) in posterior surface. The progressively increasing pattern was in dominant with clinical keratoconus, accounting for 91.7% (67/73) and 94.5% (69/73) respectively in anterior and posterior surface. Progressively increasing pattern in Orbscan was presented in 80.4% (165/196) normal eyes and suspected or clinical stage keratoconus. As to the receiver-operating-characteristic (ROC) curves and cutoff value, Orbscan I-S value (3 mm and 5 mm) of anterior and posterior surface was sensitive for the diagnosis of keratoconus suspects, with the cutoff value of anterior and posterior 5 mm I-S at 1.15 D and 0.65 D, respectively. In addition to the sensitivity of I-S values in Pentacam, the elevation value of anterior and posterior surface also displayed important diagnostic meaning. The cutoff of anterior, posterior elevation values was 4.5 µm and 7.5 µm, respectively. CONCLUSIONS: Numerous measured indicators revealed obviously difference between the two instruments in normal eyes, with slightly smaller values in Pentacam. In addition to I-S values, pattern changes and elevation values in elevation map of Pentacam represent remarkable advantages for the screening of keratoconus suspects. There is an incremental risk of corneal ectasia in the increasing pattern of elevation map.

Changes in corneal densitometry after long-term orthokeratology for myopia and short-term discontinuation.

PURPOSE: To quantify changes in corneal densitometry after long-term orthokeratology treatment in myopic children and to analyze the reversibility one month after discontinuation. METHODS: Seventy-four myopic subjects aged 8-16 years, who wore orthokeratology lenses for two years, were divided into relatively steep- (lens movement within 1.0-1.5 mm, thirty-six participants) and flat-fitting groups (lens movement within 1.5-2.0 mm, thirty-eight participants). Based on refractive errors, they were divided into low and moderate myopia groups (thirty-seven participants in each group). Corneal densitometry was performed using Pentacam (Oculus Optikgeräte GmbH, Wetzlar, Germany) at each follow-up timepoint. Repeated-measures analysis of variance was used to compare the parameters before and after orthokeratology. RESULTS: The corneal densitometry values over the 0-10 mm diameter area increased from 12.84±1.38 grayscale units (GSU) at baseline to 13.59±1.42 GSU after three-month orthokeratology (P = .001) and reached 14.92±1.45 GSU at two years (P < .001). An increase in densitometry began at one month (P = .001) over the 0-2 mm annulus compared with that at three months over the 2-6 mm and 6-10 mm zones (P = .002,.014). The densitometry values significantly increased at three months in the relatively steep-fitting group (P = .003) and at one year in the relatively flat-fitting group (P = .001). After discontinuation of orthokeratology for one month, the values showed no significant decrease. CONCLUSIONS: Long-term orthokeratology treatment causes a small but statistically significant increase in corneal densitometry values. During the first year, the onset of these changes was related to the fitting mode. Corneal densitometry values showed no significant reduction after one-month discontinuation.

Complex Interplay Between COVID-19 Lockdown and Myopic Progression.

Purpose: To compare the myopic progression before and during strict home confinement when coronavirus disease 2019 (COVID-19) outbreak and explore the potential influencing factors. Methods: A cross-sectional study. One hundred and fifteen myopic children (115 right eyes) who replace their frame-glasses from December 2019 to January 2020 and with complete refractive records in our hospital since myopia were involved in the study. At the beginning of the strict home confinement and after a 3-month strict home confinement during the COVID-19 pandemic, they were invited to our hospital to examine the axial length and refractive errors. And visual functions, convergence insufficiency symptom survey (CISS) scale and questionnaires were also performed. Besides, the axial length and refractive errors before the COVID-19 were got from outpatient case files. The effect of strict home confinement on myopia was assessed by comparing monthly axial elongation before COVID-19 and during strict home confinement. Spearman correlation analysis was performed to explore the correlation between potential influencing factors and myopia progression. Results: Axial length's monthly elongation during strict home confinement was 35% higher than normal periods (0.046 vs. 0.033 mm/month, P = 0.003). The proportion of severe asthenopia doubled (P = 0.020). For myopia progression, heredity, close indoor work time and electronic products were risk factors. Besides, the protective factors were age, rest time after continuous eye usage, sleep time and distance from eye to computer screen. Conclusions: During COVID-19, the decline in outdoor activities and increase of exposure time to digital screens accelerated the progression of myopia by 1/3.

Outcomes of different lines of keratoconus management in a tertiary eye center in north China.

AIM: To evaluate the treatment selections and outcomes of keratoconus and discuss the grading treatment of keratoconus. METHODS: Medical records of 1162 patients (1863 eyes) with keratoconus treated with rigid gas permeable (RGP), corneal collagen crosslinking, and keratoplasty were reviewed. The patients were grouped according to the CLEK Study. The advanced group was further divided into a <60 D group and >60 D group. The best-corrected visual acuity (BCVA) and topographic data before and after treatment were recorded. RESULTS: In the 761 eyes with steep K<52 D, nonsurgical management accounted for 83.4%, while in the 735 eyes with steep K>60 D, surgical management accounted for 90.6%. A total of 618 eyes had improved BCVA at the final follow-up point (>18mo, P<0.001). When steep K was <52 D, the BCVA in the RGP group was better than those with lamellar keratoplasty (LKP; P=0.028). When steep K was >52 D, the BCVA and topographic astigmatism outcomes showed no differences among the treatment groups. When steep K was >60 D, the BCVA in eyes treated with LKP was worse than those with steep K<60 D (P=0.025). The incidence of steep K progression in the RGP group was higher in advanced group (20.0% vs 10.8%, P=0.019). The probability of future keratoplasty in RGP was higher in advanced group (14.8% vs 7.0%, P=0.027). The incidence of steep K progression in the corneal collagen crosslinking (CXL) group was higher in advanced group (32.3% vs 8.5%, P=0.007). Multivariate logistic regression revealed the following related factors for treatment options: steep K [odds ratio (OR)=1.208, 95%CI: 1.052-1.387], TA (OR=1.171, 95%CI: 1.079-1.270), and TCT (OR=0.978, 95%CI: 0.971-0.984). The level of steep K, TA, and TCT all relates to the treatment choices of both keratoplasty and non-keratoplasty, while steep K provided the highest diagnostic accuracy (AUC=0.947, P<0.001). CONCLUSION: Steep K is an important grading treatment indicator. When steep K is <52 D, RGP lenses should be recommended. It is the best time for LKP when the steep K ranges from 52 to 60 D.

The therapeutic use of quercetin in ophthalmology: recent applications.

Quercetin is a natural flavonol antioxidant found in various plant sources and food samples. It is well known for its notable curative effects on the treatment of ophthalmic diseases due to various biological activities, such as antioxidant, anti-inflammatory, and anti-fibrosis activities. This review will discuss the latest developments in therapeutic quercetin for the treatment of keratoconus, Graves' orbitopathy, ocular surface, cataracts, glaucoma, retinoblastoma, and other retinal diseases.

Preparation and Evaluation of Starch Hydrogel/Contact Lens Composites as Epigallocatechin Gallate Delivery Systems for Inhibition of Bacterial Adhesion.

Microbial infections caused by wearing contact lenses has become a major health problem, so the design and development of antibacterial contact lenses has attracted widespread attention. To safely and effectively inhibit bacterial adhesion of contact lenses, we have facilely prepared epigallocatechin gallate (EGCG) loaded starch hydrogel/contact lens composites by in-situ free radical polymerization of the mixture containing 2-hydroxylethyl methacrylate, methacrylic acid and ethylene glycol dimethacrylate. The adequate transmittance of the resulting contact lenses was characterized by ultraviolet-visible spectrophotometry, and their satisfactory stability was examined using differential scanning calorimetry and thermogravimetric analysis. Whereafter, cytotoxicity and degradation experiments were performed to investigate the biocompatibility and degradability of the contact lenses. The results showed the nontoxicity and good degradability of the composites. Besides, the capacity of the contact lenses for in vitro release of EGCG was also evaluated, and the results showed that the EGCG in these contact lenses can be sustainably released for at least 14 days. Further bacterial adhesion assay suggested that the EGCG loaded starch hydrogel/contact lenses could significantly reduce the adhesion of Pseudomonas aeruginosa compared to the control. The EGCG loaded starch hydrogel/contact lens composites provide a potential intervention strategy for preventing ocular microbial infections and inhibiting bacterial keratitis.

Gelatin hydrogel/contact lens composites as rutin delivery systems for promoting corneal wound healing.

Corneal wound healing is a highly regulated biological process that is of importance for reducing the risk of blinding corneal infections and inflammations. Traditional eye drop was the main approach for promoting corneal wound healing. However, its low bioavailability required a high therapeutic concentration, which can lead to ocular or even systemic side effects. To develop a safe and effective method for treating corneal injury, we fabricated rutin-encapsulated gelatin hydrogel/contact lens composites by dual crosslinking reactions including in situ free radical polymerization and carboxymethyl cellulose/N-hydroxysulfosuccinimide crosslinking. In vitro drug release results evidenced that rutin in the composites could be sustainedly released for up to 14 days. In addition, biocompatibility assay indicated nontoxicity of the composites. Finally, the effect of rutin-encapsulated composites on the healing of the corneal injury in rabbits was investigated. The injury was basically cured in corneas using rutin-encapsulated composites (healing rate, 98.3% ± 0.7%) at 48 h post-operation, while the damage was still present in corneas using the composite (healing rate, 87.0% ± 4.5%). Further proteomics analysis revealed that corneal wound healing may be promoted by the ERK/MAPK and PI3K/AKT signal pathways. These results inform a potential intervention strategy to facilitate corneal wound healing in humans.

Reservoir quality and its controlling diagenetic factors in the Bentiu Formation, Northeastern Muglad Basin, Sudan.

The Abu Gabra and Bentiu formations are widely distributed within the interior Muglad Basin. Recently, much attention has been paid to study, evaluate and characterize the Abu Gabra Formation as a proven reservoir in Muglad Basin. However, few studies have been documented on the Bentiu Formation which is the main oil/gas reservoir within the basin. Therefore, 33 core samples of the Great Moga and Keyi oilfields (NE Muglad Basin) were selected to characterize the Bentiu Formation reservoir using sedimentological and petrophysical analyses. The aim of the study is to de-risk exploration activities and improve success rate. Compositional and textural analyses revealed two main facies groups: coarse to-medium grained sandstone (braided channel deposits) and fine grained sandstone (floodplain and crevasse splay channel deposits). The coarse to-medium grained sandstone has porosity and permeability values within the range of 19.6% to 32.0% and 1825.6 mD to 8358.0 mD respectively. On the other hand, the fine grained clay-rich facies displays poor reservoir quality as indicated by porosity and permeability ranging from 1.0 to 6.0% and 2.5 to 10.0 mD respectively. A number of varied processes were identified controlling the reservoir quality of the studies samples. Porosity and permeability were enhanced by the dissolution of feldspars and micas, while presence of detrital clays, kaolinite precipitation, iron oxides precipitation, siderite, quartz overgrowths and pyrite cement played negative role on the reservoir quality. Intensity of the observed quartz overgrowth increases with burial depth. At great depths, a variability in grain contact types are recorded suggesting conditions of moderate to-high compactions. Furthermore, scanning electron microscopy revealed presence of micropores which have the tendency of affecting the fluid flow properties in the Bentiu Formation sandstone. These evidences indicate that the Bentiu Formation petroleum reservoir quality is primarily inhibited by grain size, total clay content, compaction and cementation. Thus, special attention should be paid to these inhibiting factors to reduce risk in petroleum exploration within the area.

Extended Contact Lens Wear Promotes Corneal Norepinephrine Secretion and Pseudomonas aeruginosa Infection in Mice.

Purpose: Extended contact lens (CL) wear predisposes the wearer to Pseudomonas aeruginosa infection of the cornea, but the mechanism involved remains incompletely understood. The purpose of this study was to investigate the role of the stress hormone norepinephrine (NE) in the pathogenesis of CL-induced P. aeruginosa keratitis. Methods: A total 195 adult C57BL/6 mice were used in this study. Corneal NE content was measured after 48 hours of sterile CL wear in mice. The effect of NE on P. aeruginosa adhesion and biofilm formation on the CL surface was examined in vitro. Moreover, mouse eyes were covered with P. aeruginosa-contaminated CLs, and either 500-µM NE was topically applied or the eyes were subconjunctivally injected with 100 µg of N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) to deplete local NE. Clinical scores, neutrophil infiltration, proinflammatory cytokine levels, and bacterial load on the corneas and CLs were evaluated. Results: Corneal NE content was elevated with extended CL wear in mice. In vitro, NE promoted the adhesion and biofilm formation of P. aeruginosa on the CL surface. In mice, topical application of NE aggravated P. aeruginosa infection, accompanied with increased clinical scores, neutrophil infiltration, proinflammatory cytokine expression, and bacterial burden on the corneas and CLs. However, pre-depletion of local NE with DSP-4 significantly alleviated the severity of P. aeruginosa keratitis. Conclusions: Extended CL wear elevates corneal NE content, which promotes the pathogenesis of CL-induced P. aeruginosa keratitis in mice. Targeting NE may provide a potential strategy for the treatment of CL-related corneal infection caused by P. aeruginosa.

Comparison of long-term results after manual and femtosecond assisted corneal trephination in deep anterior lamellar keratoplasty for keratoconus.

AIM: To compare long-term postoperative outcomes of manual and femtosecond assisted corneal trephination in deep anterior lamellar keratoplasty (FS-DALK) for keratoconus. METHODS: In the retrospective study, 17 consecutive eyes that underwent vertical side cut incision FS-DALK and 22 eyes that underwent trephine incision DALK were collected over a 2-year period. Main measurements included postoperative uncorrected-visual acuity (UCVA), corrected distance visual acuity (CDVA), refractive sphere and cylinder, manifest refraction spherical equivalent (MRSE), flat and steep corneal keratometry (K1 and K2), endothelial cell density (ECD), and time of epithelium healing and suture removal. RESULTS: Groups were comparable for diagnosis and preoperative visual acuity. Follow-up averaged 23mo (range, 12-36mo). At 12mo, the mean UCVA was better in the manual-DALK group (P=0.039), and the refractive sphere was lower in the FS-DALK group (P=0.040). MRSE between groups differed at 1, 6, and 12mo postoperatively (P=0.047, 0.025, 0.042, respectively). Mean CDVA, cylinder, K1, K2, corneal astigmatism, ECD, and time of epithelium healing were similar between groups. Stability of MRSE, ECD, and K1 returned sooner after FS-DALK. Initial loosened suture removal time was earlier in the manual-DALK group (P=0.042) while complete suture removal time was similar (P=0.122). CONCLUSION: Manual and femtosecond assisted corneal trephination in DALK are options for advanced keratoconus. FS-DALK do not result in improved visual acuity but it is more stable during the follow-up period. FS-DALK in the present form show limited benefit, so surgical design and parameters still need to be optimized and explored.

MAVS O-GlcNAcylation Is Essential for Host Antiviral Immunity against Lethal RNA Viruses.

It is known that lethal viruses profoundly manipulate host metabolism, but how the metabolism alternation affects the immediate host antiviral immunity remains elusive. Here, we report that the O-GlcNAcylation of mitochondrial antiviral-signaling protein (MAVS), a key mediator of interferon signaling, is a critical regulation to activate the host innate immunity against RNA viruses. We show that O-GlcNAcylation depletion in myeloid cells renders the host more susceptible to virus infection both in vitro and in vivo. Mechanistically, we demonstrate that MAVS O-GlcNAcylation is required for virus-induced MAVS K63-linked ubiquitination, thereby facilitating IRF3 activation and IFNß production. We further demonstrate that D-glucosamine, a commonly used dietary supplement, effectively protects mice against a range of lethal RNA viruses, including human influenza virus. Our study highlights a critical role of O-GlcNAcylation in regulating host antiviral immunity and validates D-glucosamine as a potential therapeutic for virus infections.