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Infectious uveitis is a vision-threatening condition that requires prompt clinical diagnosis and proper treatment. However, rapid and proper diagnosis in infectious uveitis remains challenging. Several examination tests, including polymerase chain reaction (PCR) tests, are transitioning from laboratory-based basic research-level tests to bedside clinical tests, and recently tests have changed to where they can be performed right next to clinicians. In this review, we introduce an updated overview of recent studies that are representative of the current trends in clinical microbiological techniques including PCR tests for infectious uveitis.

Rapid diagnostics for Chlamydia trachomatis and Neisseria gonorrhoeae are desirable so that patients can be treated while they are still in the clinic or doctor's office. The Cepheid GeneXpert® (Xpert) CT/NG assay was US FDA-cleared in December 2012. The assay is a rapid real-time PCR nucleic acid amplified test. The cartridge-based assay detects DNA of Chlamydia trachomatis and Neisseria gonorrhoeae. It is FDA-cleared for use in female endocervical swabs, patient-collected vaginal swabs and for female and male urine specimens from symptomatic and asymptomatic patients. It has demonstrated near-perfect sensitivity and specificity in urogenital specimens. The Xpert is a modular platform for testing samples directly from patients, which requires no hands-on manipulation from specimen loading until results are available. Results are provided in approximately 90 minutes. It has been graded by the FDA as moderately complex for Clinical Laboratory Improvement Amendments. Several publications have reported its promising use in clinical settings.

Background Fungal species are responsible for 40%–50% of all microbial keratitis cases. Due to the low amount of extracted DNA in ocular Formalin‐fixed Paraffin‐embedded (FFPE) samples, selecting a reliable molecular method is a substantial issue in this field. Methods Sixty‐six samples were collected via the penetrating keratoplasty (PK) technique. Histopathology assays were performed using hematoxylin–eosin (H&E) and periodic acid Schiff (PAS) staining methods. The ITS1/ITS4 and ITS1/ITS2 primer pairs were used in a semi‐nested polymerase chain reaction (PCR) to target the universal internal transcribed spacer (ITS) region. Some PCR results were validated through sequencing. Results Fungal DNA was detected in 44 of 66 samples (66.7%), and histopathology was positive for 41 of 66 samples (62.1%). Of 41 histopathologically proven fungal‐positive cases, 39 were PCR‐positive (95%). Moreover, of 44 PCR‐positive samples, 39 (88.6%) were histopathology‐positive, and 5 (11.3%) were histopathology‐negative. Totally in 39 cases (59%), both histopathology and PCR yielded positive results. The Kappa agreement rate between the two diagnostic methods, including histopathology and PCR, was 0.77. Sensitivity, specificity, positive predictive value, and false predictive value were reported as 88.64%, 90.9%, 95.12%, and 80%, respectively. Conclusion As we reached the acceptable Kappa agreement rate, we concluded that applying the semi‐nested PCR assay is a promising method for supporting the evidence by histopathology. Finally, we suggest targeting more specific gene regions using primer pairs that amplify smaller amplicon sizes and surveying novel molecular methods such as NGS to achieve higher sensitivity and Kappa agreement rates. Following the clinical suspicion for fungal keratitis (FK), the corneal samples were collected by penetrating keratoplasty (PK) from the patients. Specimens were prepared by microtome and were stained by hematoxylin and eosin (H&E), periodic acid schiff (PAS) stainings. For the molecular assay, samples were subjected to DNA extraction and histopathology examination. The amplification of the β‐globin gene by conventional PCR was used to confirm the quality of extracted DNA. The semi‐nested PCR was performed using ITS1, ITS2, and ITS4 primers during two steps. Sequencing the internal transcribed spacer region (ITS1‐5.8 S‐ITS2) to identify causative agents was performed on PCR products. The results were analyzed by researchers.

An isocratic reversed-phase HPLC method with ultraviolet detection at 205 nm has been developed for analysis of cyclosporine A (CyA) in rabbit ocular samples. Neither internal standard nor extraction was needed for sample preparation. Acetonitrile (ACN; 1 mL) was added to 250 μL aqueous and vitreous samples to precipitate proteins. The supernatant was dried and the residue was reconstituted in mobile phase and injected for HPLC analysis. Chromatography was performed on an octadecyl silane-A (ODSA) C18 (4.6 × 250 mm, 5 μm) column. The column temperature was fixed at 70 °C and the mobile phase was ACN 65%, methanol 20% and water 15% at a flow rate of 1.5 mL min−1. The calibration curve for CyA in rabbit ocular samples was linear over the concentration range 0.2 and 10 μg mL−1 with a correlation coefficient of 0.9992. Intra-day and inter-day precision were 4.61–7.83% and 5.27–10.70%, respectively. Intra-day and inter-day accuracy were 89.2–108% and 83.4–111%, respectively. The limits of detection (LOD) and quantification (LOQ) were 5.7 and 38 ng mL−1, respectively. The method was successfully used for analysis of CyA in real aqueous and vitreous humor samples from New Zealand albino rabbits. The method is therefore suitable for analysis of CyA in ocular samples.

Publication bias is a serious problem in systematic reviews and meta-analyses, which can affect the validity and generalization of conclusions. Currently, approaches to dealing with publication bias can be distinguished into two classes: selection models and funnel-plot-based methods. Selection models use weight functions to adjust the overall effect size estimate and are usually employed as sensitivity analyses to assess the potential impact of publication bias. Punnel-plot-based methods include visual examination of a funnel plot, regression and rank tests, and the nonparametric trim and fill method. Although these approaches have been widely used in applications, measures for quantifying publication bias are seldom studied in the literature. Such measures can be used as a characteristic of a meta-analysis; also, they permit comparisons of publication biases between different meta-analyses. Egger's regression intercept may be considered as a candidate measure, but it lacks an intuitive interpretation. This article introduces a new measure, the skewness of the standardized deviates, to quantify publication bias. This measure describes the asymmetry of the collected studies' distribution. In addition, a new test for publication bias is derived based on the skewness. Large sample properties of the new measure are studied, and its performance is illustrated using simulations and three case studies.

To compare two methods of surgical augmentation (prism adaptation and the augmented surgery formula) in the management of acquired comitant esotropia. Forty patients were included in this prospective study and assigned to either the prism adaptation (20 patients) or augmented surgery (20 patients) group. After preoperative prism adaptation, patients in the prism adaptation group were classified as prism adaptation responders (fusers) or non-responders (non-fusers). All patients in the prism adaptation group underwent surgery for the prism-adapted angle. Patients in the augmented surgery group underwent surgery based on the augmented surgery formula, defined as the average of the near deviation without correction and the distance deviation with correction. In the prism adaptation group, 6 patients (30%) were prism responders, whereas 14 (70%) were non-responders. The 3-month motor success rate was significantly higher in the prism adaptation group (90%) than the augmented surgery group (55%) (P = .013). The 6-month motor success rate was not significantly higher in the prism adaptation group (95%) than the augmented surgery group (80%) (P = .151). The improvement in the 6-month outcome was mainly attributed to hyperopic spectacle power reduction after 3-month postoperative evaluation to correct consecutive exotropia. Although prism adaptation is superior in precisely determining the surgical target angle, the success rates were comparable between the two groups after hyperopic spectacle power reduction. This obviates the need for prism adaptation, except in cases of non-accommodative esotropia. To maximize the benefit of prism adaptation, it is recommended that all patients with prism adaptation (responders and non-responders) undergo surgery for the prism-adapted angle. [J Pediatr Ophthalmol Strabismus. 2020;57(2):108-119.].

Background Myopia is prevalent in China; however, trials involving Chinese children wearing dual-focus soft contact lenses (DFSCL) are limited. Thus, the purpose of this study is to investigate the efficacy of DFSCL among Chinese school-age children. Methods Sixty-four children aged 8–12 years with spherical equivalent refraction (SER) between − 0.75D and − 4.00D were recruited in this randomized controlled clinical study. The control group (32 subjects) wore single-vision spectacles (SVS), while the DFSCL group (32 subjects) wore daily disposable + 2.00 D defocus MiSight DFSCL. Follow-up examinations were performed every 3 months to compare the axial length (AL) growth and SER change between the groups for a period of 12 months by using the independent samples t -test or the Mann-Whitney U test. Statistical differences with a P  < 0.05, when compared to the control group, are considered indicative of an effective intervention. Multivariate analysis and regression analysis were used to eliminate the effects of confounding factors on the results. Results A total of 58 subjects, with 30 in the SVS group and 28 in the DFSCL group, completed the follow-up. After adjusting for baseline age, gender, AL and SER, AL growth was 0.33 ± 0.02 mm in the SVS group and 0.23 ± 0.03 mm in the DFSCL group ( P  = 0.004). SER change was − 0.53 ± 0.06 in the SVS group and − 0.44 ± 0.06 in the DFSCL group ( P  = 0.308). In the DFSCL group, AL and SER increased 0.11 mm and 0.09 D less than in the SVS group, respectively. Moreover, initial wear of DFSCL may cause occasional blurriness in near vision, and prolonged wear may lead to increased ocular discomfort symptoms such as dryness, itchiness, and foreign body sensation. Conclusion MiSight DFSCL showed a reduction in AL growth during the first three months of wear. However, no significant benefits were observed during the subsequent nine months. No significant differences in the changes of SER were found. Trial registration Chinese Clinical Trial Registry: ChiCTR2200064731. Registered 15 October 2022, http://www.chictr.org.cn/.

Over the last two decades, a large number of non-communicable/chronic disorders reached an epidemic level on a global scale such as diabetes mellitus type 2, cardio-vascular disease, several types of malignancies, neurological and eye pathologies—all exerted system’s enormous socio-economic burden to primary, secondary, and tertiary healthcare. The paradigm change from reactive to predictive, preventive, and personalized medicine (3PM/PPPM) has been declared as an essential transformation of the overall healthcare approach to benefit the patient and society at large. To this end, specific biomarker panels are instrumental for a cost-effective predictive approach of individualized prevention and treatments tailored to the person. The source of biomarkers is crucial for specificity and reliability of diagnostic tests and treatment targets. Furthermore, any diagnostic approach preferentially should be noninvasive to increase availability of the biomaterial, and to decrease risks of potential complications as well as concomitant costs. These requirements are clearly fulfilled by tear fluid, which represents a precious source of biomarker panels. The well-justified principle of a “sick eye in a sick body” makes comprehensive tear fluid biomarker profiling highly relevant not only for diagnostics of eye pathologies but also for prediction, prognosis, and treatment monitoring of systemic diseases. One prominent example is the Sicca syndrome linked to a cascade of severe complications that include dry eye, neurologic, and oncologic diseases. In this review, protein profiles in tear fluid are highlighted and corresponding biomarkers are exemplified for several relevant pathologies, including dry eye disease, diabetic retinopathy, cancers, and neurological disorders. Corresponding analytical approaches such as sample pre-processing, differential proteomics, electrophoretic techniques, high-performance liquid chromatography (HPLC), enzyme-linked immuno-sorbent assay (ELISA), microarrays, and mass spectrometry (MS) methodology are detailed. Consequently, we proposed the overall strategies based on the tear fluid biomarkers application for 3P medicine practice. In the context of 3P medicine, tear fluid analytical pathways are considered to predict disease development, to target preventive measures, and to create treatment algorithms tailored to individual patient profiles.

Background Toxoplasma gondii ( T. gondii ) is one of the most common protozoan parasites distributed worldwide. This parasite causes different clinical manifestations including ocular toxoplasmosis, which presents as chorioretinitis, vision impairment, and even blindness. Accurate determination of the different genotypes of T. gondii contributing to various infection types is of great importance, especially in cases of ocular infection. This study aimed to determine the genotypes of Toxoplasma parasite isolated for the first time from the eyes of patients with ocular toxoplasmosis in Iran. Methods A total of 44 aqueous humor and/or vitreous fluid samples were collected in hospital operating rooms from patients with ocular toxoplasmosis and used for genotype analysis. After genomic DNA extraction, two regions at the 5’ and 3’ ends of the Toxoplasma SAG2 gene were amplified and digested by the restriction endonucleases Sau3aI and HhaI , respectively. Results Amplification of the SAG2 locus was successful in 38 samples. Nested PCR of the 5’ and 3’ ends of this locus produced fragments of approximately 242 bp and 222 bp, respectively. According to the target restriction maps, 37 samples (97.37%) were identified as type I, and one sample was type II (2.63%). Conclusions This study is the first report of direct genotyping of intra-ocular fluid specimens from Iranian patients with OT that revealed a striking predominance of T. gondii type I. These findings indicate the potential role of this highly virulent lineage of the parasite in severe ocular disease in Iran, and suggest that appropriate management and control strategies may be required in this population.

Background: The utilities of anterior segment optical coherence tomography (AS-OCT) for characterization, differential diagnosis, postoperative monitoring, and evaluation/comparison of surgical techniques in pterygium are described. Through AS-OCT, it is also possible to study the corneal astigmatic effect of pterygium. Our purpose is to study the associations between the anatomical characteristics of pterygium and the corneal topography through AS-OCT. Methods: Fifty eyes with primary pterygium in a tertiary hospital were evaluated before surgery by measuring 10 anatomical variables of pterygium and 13 topographic variables using AS-OCT (Casia 2; Tomey Corp., Nagoya, Japan). Statistical analysis was used to study the association between them. Results: Pterygium classified as flat pattern exhibited lower preoperative values of flat keratometry (K1), real flat keratometry (K1r), average keratometry (AvgK), and real average keratometry (AvgKr) compared to nodular ones. The flat pattern showed greater cylinder (CYL) and real cylinder (CLYr) values. The horizontal corneal invasion proportionally increased CYL and CYLr. Overall, larger anatomical pterygium measurements (limbus thickness (LimbusT), central pterygium thickness (CentreT), head pterygium thickness (HeadT), epithelial thickness at 1 mm (EpitT1mm), stromal thickness at 1 mm (stromT1mm), total thickness at 1 mm (TotalT1mm), total thickness at 2 mm (TotalT2mm), and total thickness at 3 mm (TotalT3mm)) resulted in lower anterior K1, K1r, AvgK, and AvgKr, and posterior K1 and AvgK values. CentreT was greater in astigmatisms against the rule than in oblique ones. Conclusions: This study demonstrates associations between preoperative topography and the NF (nodular or flat) classification of pterygium and its anatomical measurements assessed by AS-OCT.