Assessing neophyte response to daily disposable silicone hydrogel contact lenses: A randomised clinical trial investigation over one month.

OBJECTIVE: This randomised clinical trial assessed the impact on symptoms, tear film dynamics and ocular surface integrity of daily disposable silicone-hydrogel contact lenses (CLs) over a month, paying special attention to lid wiper epitheliopathy (LWE) and its implications for CL discomfort. METHODS: Neophyte CL wearers (n = 44, 21.09 ± 5.00 years old) were randomly assigned to either the experimental (n = 24) or control group (n = 20). Participants assigned to the experimental group were required to wear daily disposable CLs for 1 month for at least 8 h/day and 6 days/week. All participants were healthy subjects (no history of ocular surgery or active ocular disease) with spherical refractive errors between -8.00 and +5.00 D and cylindrical power <0.75 D. At the baseline and 1-month sessions, the Dry Eye Questionnaire 5 (DEQ-5) was completed, together with the measurement of tear film osmolarity with the TearLab osmometer, tear meniscus height (TMH) and lipid layer pattern (LLP) using a slit-lamp with Tearscope Plus attached, fluorescein break-up time (FBUT), maximum blink interval (MBI), corneal staining with fluorescein under cobalt blue light and LWE with lissamine green under slit lamp and halogen white light. RESULTS: At the baseline session, LWE showed a negative correlation with DEQ-5 (r = -0.37, p = 0.02). Significant differences in FBUT and LWE (p = 0.04) and a positive correlation between LWE and DEQ-5 (r = 0.49, p = 0.007) were observed at 1 month. Intrasession analysis at 1 month showed significant differences between the experimental and control groups in DEQ-5, FBUT and LWE (all p ≤ 0.02). Intersession analysis in the experimental group showed variations in DEQ-5, FBUT and LWE (all p ≤ 0.02) but no significant variation in the control group (all p ≥ 0.11). CONCLUSION: The presence of LWE was significantly correlated with higher symptom values in the DEQ-5. Also, participants in the experimental group presented higher values of LWE after 1 month of CL wear, in comparison with the control group.

Meibum Lipidomic Analysis in Evaporative Dry Eye Subjects.

Meibomian Glands (MG) are sebaceous glands responsible for the production of meibum, the main component of the Tear Film Lipid Layer (TFLL). The TFLL facilitates the spread of the tear film over the ocular surface, provides stability and reduces tear evaporation. Alterations in meibum composition lead to different ocular alterations like Meibomian Gland Dysfunction (MGD) and subsequent Evaporative Dry Eye (EDE). The aim of the present study was to investigate the composition and abundance of meibum lipids and their relationship with eyelid margin abnormalities, lipid layer patterns and MG status. The study utilizes a lipidomic approach to identify and quantify lipids in meibum samples using an Elute UHPLC system. This system considered all four dimensions (mass/charge, retention time, ion mobility and intensity) to provide the accurate identification of lipid species. Samples were categorized as healthy or low/no signs of alteration (group 1) or severe signs of alteration or EDE/MGD (group 2). The current investigation found differences in Variable Importance in Projection lipid abundance between both groups for the MGD signs studied. Changes in meibum composition occur and are related to higher scores in eyelid margin hyperaemia, eyelid margin irregularity, MG orifice plugging, MG loss and lipid layer pattern.

Phospho-RNA-seq: a modified small RNA-seq method that reveals circulating mRNA and lncRNA fragments as potential biomarkers in human plasma.

Extracellular RNAs (exRNAs) in biofluids have attracted great interest as potential biomarkers. Although extracellular microRNAs in blood plasma are extensively characterized, extracellular messenger RNA (mRNA) and long non-coding RNA (lncRNA) studies are limited. We report that plasma contains fragmented mRNAs and lncRNAs that are missed by standard small RNA-seq protocols due to lack of 5' phosphate or presence of 3' phosphate. These fragments were revealed using a modified protocol ("phospho-RNA-seq") incorporating RNA treatment with T4-polynucleotide kinase, which we compared with standard small RNA-seq for sequencing synthetic RNAs with varied 5' and 3' ends, as well as human plasma exRNA Analyzing phospho-RNA-seq data using a custom, high-stringency bioinformatic pipeline, we identified mRNA/lncRNA transcriptome fingerprints in plasma, including tissue-specific gene sets. In a longitudinal study of hematopoietic stem cell transplant patients, bone marrow- and liver-enriched exRNA genes were tracked with bone marrow recovery and liver injury, respectively, providing proof-of-concept validation as a biomarker approach. By enabling access to an unexplored realm of mRNA and lncRNA fragments, phospho-RNA-seq opens up new possibilities for plasma transcriptomic biomarker development.

Comparison Between Ocular Biometric Parameters and Intraocular Pressure With and Without Contact Lenses.

OBJECTIVES: To compare the values of central corneal thickness, anterior chamber depth (ACD), lens thickness (LT), vitreous length (VL), axial length (AL), and corneal-compensated intraocular pressure performed with and without contact lenses (CL). METHODS: Forty volunteer participants (16 men and 24 women, 24.2±2.9 years) were recruited. In a single visit, participants underwent autorefraction, keratometry, biometry, topography, pachymetry, and tonometry with the naked eye (without CL). Then, biometry, pachymetry, and tonometry were repeated twice wearing two CL (somofilcon A and nesofilcon A) fitted in a random order. RESULTS: Many of the ocular biometric values were affected by the use of CL during measurements (paired t test; all P ≤0.003), except for LT and VL (Wilcoxon test; both P ≥0.272). Corneal-compensated intraocular pressure was also affected by contact lens wear during measurements, obtaining lower values when wearing them (Wilcoxon test; all P ≤0.001). CONCLUSIONS: Central corneal thickness, ACD, AL, and corneal-compensated intraocular pressure measurements cannot be performed while wearing CL. However, LT and VL measurements were not affected by any contact lens use. In addition, it was observed that ACD results from both devices are not interchangeable either when measured with the naked eye or using any contact lens.

Meibomian gland secretion quality association with ocular parameters in university students during COVID- 19 restrictions.

PURPOSE: To determine if the Meibomian Gland (MG) secretion quality is associated with symptoms of ocular discomfort, hours of Video Display Terminals (VDT) use, eyelid margin abnormalities, conjunctival hyperemia, and Meibomian Gland Loss Area (MGLA) in a sample of university students. METHODS: An online survey that included an Ocular Surface Disease Index (OSDI) questionnaire and an extra question about hours of VDT use recruited an initial sample of 183 participants. Only 120 participants that fulfilled the inclusion criteria were scheduled for a battery of ocular surface and MG specific exam. The tests include: 1) meibometry, 2) slit lamp exploration of eyelid margin abnormalities (irregularity, hyperemia and MG orifices plugging), MG secretion quality and conjunctival hyperemia, and 3) Meibography. RESULTS: Significant positive correlations between the MG secretion quality and eyelid margin hyperemia, MG orifices plugging, MGLA, nasal conjunctival hyperemia, and temporal conjunctival hyperemia (Spearman Rho; all r>0.186, p<0.042) were found. Multivariate regression found association between OSDI with hours of VDT use (B=0.316, p=0.007), and eyelid hyperemia (B=0.434, p≤ 0.001). A statistical association between MG secretion quality and eyelid margin hyperemia, MG orifices plugging, MGLA and conjunctival hyperemia (Fisher's exact; all p<0.039) were found. Multivariate regression found association between MG secretion quality with MG orifices plugging (B=0.295, p=0.004) and meibometry (B=-0.001, p=0.029). CONCLUSION: Participants with higher values in MG secretion quality have higher values in eyelid margin hyperemia, MG plugging, MGLA, and conjunctival hyperemia. No direct relationship between MG secretion quality and hours of VDT use or OSDI were found.

Validation of a New Software Application for Tear Breakup Measurement.

SIGNIFICANCE: Tear film instability has been recognized as one key mechanism on dry eye disease. There is a need for new instruments, methods, or protocols to improve the repeatability and reproducibility of the tear film stability to facilitate its clinical evaluation use as a rapid and reliable primary diagnostic test. PURPOSE: The present pilot study aimed to validate a method to automatically measure the tear film breakup time (BUT). METHODS: A total of 264 videos of BUT were randomly selected among the clinical history of subjects attending the optometry clinic. Videos were stored in a backup server through a secure file transfer protocol and analyzed by three different examiners: two masked observers (subjective evaluation) and a third investigator using the automatic software application (objective evaluation). Subjective evaluation was conducted only once on an online software designed for this protocol where videos were presented in random masked order. Automatic evaluation based on color and texture analysis was performed by (1) automatic localization of sequences of interest in the video, (2) extraction of the region of interest within each frame, and (3) automatic BUT measurement from evolution curves in the region of interest as time elapsed from the beginning of the sequence of interest until the curve exceeds a threshold. RESULTS: Substantial correlation was observed among the examiners (intraclass correlation coefficient, 0.752). There was a statistical difference between observer 1 and 2 evaluations (t test, P < .001), whereas data provided by the software showed no significant differences from those of the observers (t test, P ≥ .26). Similar results to the whole data set analysis were obtained when the sample was reassessed only considering mean BUT values ≤15 seconds. CONCLUSIONS: The present pilot study showed acceptable clinical results for the software application designed to objectively measure the BUT.

Release of Cell-free MicroRNA Tumor Biomarkers into the Blood Circulation with Pulsed Focused Ultrasound: A Noninvasive, Anatomically Localized, Molecular Liquid Biopsy.

Purpose To compare the abilities of three pulsed focused ultrasound regimes (that cause tissue liquefaction, permeabilization, or mild heating) to release tumor-derived microRNA into the circulation in vivo and to evaluate release dynamics. Materials and Methods All rat experiments were approved by the University of Washington Institutional Animal Care and Use Committee. Reverse-transcription quantitative polymerase chain reaction array profiling was used to identify candidate microRNA biomarkers in a rat solid tumor cell line. Rats subcutaneously grafted with these cells were randomly assigned among three pulsed focused ultrasound treatment groups: (a) local tissue liquefaction via boiling histotripsy, (b) tissue permeabilization via inertial cavitation, and (c) mild (<10°C) heating of tissue, as well as a sham-treated control group. Blood specimens were drawn immediately prior to treatment and serially over 24 hours afterward. Plasma microRNA was quantified with reverse-transcription quantitative polymerase chain reaction, and statistical significance was determined with one-way analysis of variance (Kruskal-Wallis and Friedman tests), followed by the Dunn multiple-comparisons test. Results After tissue liquefaction and cavitation treatments (but not mild heating), plasma quantities of candidate biomarkers increased significantly (P value range, <.0001 to .04) relative to sham-treated controls. A threefold to 32-fold increase occurred within 15 minutes after initiation of pulsed focused ultrasound tumor treatment, and these increases persisted for 3 hours. Histologic examination confirmed complete liquefaction of the targeted tumor area with boiling histotripsy, in addition to areas of petechial hemorrhage and tissue disruption by means of cavitation-based treatment. Conclusion Mechanical tumor tissue disruption with pulsed focused ultrasound-induced bubble activity significantly increases the plasma abundance of tumor-derived microRNA rapidly after treatment. © RSNA, 2016 Online supplemental material is available for this article.

Accuracy of a Freezing Point Depression Technique Osmometer.

PURPOSE: The purpose of this study was to examine the precision and accuracy of the Fiske 110 Osmolarity System under different protocols to determine the possible applications of this device in tear film research and clinical practice. METHODS: Three separate studies were performed. In the first, Fiske 110 measurements were made on undiluted and diluted (1:1, 1:4, and 1:9 dilutions) standard samples of different osmolarity values: 50, 290, and 850 mOsm/kg and 297 and 338 mOsm/L. In the second study, measurements were made on different types of contact lens care solutions. Finally, in an agreement study, measurements were made in two sets of 60 subjects to compare TearLab versus Fiske 110 (using both 2- and 4-µL tear sample). RESULTS: Although osmolarity measurements for undiluted solutions differed statistically from reference standard values, all biases were in the tolerance range proposed by the manufacturer except for the 850-mOsm/kg solution. No significant differences from reference osmolarity values were observed for the 1:1 and 1:4 diluted 297- and 338-mOsm/L H2O solutions, respectively, although all diluted solutions showed a possible bias out of the range provided. Osmolarities for the soft contact lens solutions fell within the range 293 to 309 mOsm/kg with the exception of Opti-Free Express (225 mOsm/kg). In the agreement study, significant differences were observed between measurements obtained using the TearLab and both Fiske 110 procedures, although the Fiske 110 (4 µL) procedure was closer to the TearLab than the Fiske (2 µL) procedure. CONCLUSIONS: For undiluted solutions, the Fiske 110 shows good performance, making it a useful device for osmolarity measurements in lens care solutions or eye drops. A worse performance was observed for more diluted standard solution samples. When testing diluted samples, performance was acceptable for osmolarity values close to tear values.

Correlation between tear osmolarity and tear meniscus.

PURPOSE: To examine the relationship between tear meniscus height (TMH) and subjective meniscus grading (subjective tear meniscus [TM]) with tear osmolarity. METHODS: Tear osmolarity measurements (using TearLab) and digital images of the TM were obtained in 177 consecutive patients undergoing an eye examination at our optometry clinic (Universidad de Santiago de Compostela, Spain) who fulfilled the study's inclusion criteria. Participants were also administered the McMonnies and Ocular Surface Disease Index questionnaires for the detection of dry eye disease. The lower TM was videotaped by a digital camera attached to a slit lamp in its central portion without fluorescein instillation. After the study, a masked observer extracted an image from each video and measured the TMH using open source software (NIH ImageJ). Subsequently, the masked observer subjectively graded the appearance of each meniscus. For statistical analysis, subjects were stratified by age and by dry eye symptoms as indicated by their scores in the two questionnaires. RESULTS: In the whole study population, a significant relationship was observed between osmolarity and TMH (r = -0.41, p < 0.001) and osmolarity and subjective TM (r = 0.35, p < 0.001). A cluster analysis revealed similar correlations when subjects were stratified by age or dry eye symptoms, these correlations being more pronounced in older and more symptomatic subjects. Objective TMH measurements and subjective meniscus quality were also correlated (r = -0.75, p < 0.001). CONCLUSIONS: Osmolarity and both objective TMH measurements and subjective interpretation of the meniscus showed high correlation, especially in older symptomatic subjects.

Intraocular pressure fluctuations assessment in professional wind instrument players.

CLINICAL RELEVANCE: Due to the long-time that wind musicians spend playing their instruments, it is important to investigate if intraocular pressure could be affected by this activity. BACKGROUND: To assess the intraocular pressure fluctuations and fluctuations affecting factors in professional wind musicians while playing different tones. METHODS: Thirty professional wind musicians (23.0 ± 3.20 years) were recruited from the Professional Music College of A Coruña. A questionnaire about environmental/demographic factors was given to participants. Intraocular pressure was measured four times by ICare IC100 tonometer: before, during low and high-pitched tones, and immediately after stopping playing the wind instrument. RESULTS: Pairwise comparison revealed statistical differences between measurement points (Sidak, all p ≤ 0.019), except between before playing and while playing low-pitched tones (Sidak, p = 1.000). Intraocular pressure increases during high pitch playing and decreases after stopping playing. No significant differences in intraocular pressure fluctuation were reported between physically active (>2 days/week) and non-physically active participants (Unpaired t-test, p = 0.680). All intraocular pressure values were positively correlated (Pearson's correlation, all r ≥ 0.505, p ≤ 0.004). Intraocular pressure fluctuations were negatively correlated with musical playing years (Pearson's correlation, r = - 0.396, p = 0.030). There were no significant correlations among intraocular pressure fluctuation and gender, age, weight, height, or daily time playing (Pearson's correlation, all p ≥ 0.058). CONCLUSION: Professional wind musicians suffer intraocular pressure peaks while playing high-pitched tones; therefore, ocular fundus evaluation and visual campimetry should be performed as routine tests in the visual exam of this population.

Unveiling a Technosol-based remediation approach for enhancing plant growth in an iron-rich acidic mine soil from the Rio Tinto Mars analog site.

This paper explores the potential of Technosols made from non-hazardous industrial wastes as a sustainable solution for highly acidic iron-rich soils at the Rio Tinto mining site (Spain), a terrestrial Mars analog. These mine soils exhibit extreme acidity (pHH2O = 2.1-3.0), low nutrient availability (non-acid cation saturation < 20 %), and high levels of Pb (3420 mg kg-1), Cu (504 mg kg-1), Zn (415 mg kg-1), and As (319 mg kg-1), hindering plant growth and ecosystem restoration. To address these challenges, the study systematically analyzed selected waste materials, formulated them into Technosols, and conducted a four-month pot trial to evaluate the growth of Brassica juncea under greenhouse conditions. Technosols were tailored by adding varying weight percentages of waste amendments into the mine Technosol, specifically 10 %, 25 %, and 50 %. The waste amendments comprised a blend of organic waste (water clarification sludge, WCS) and inorganic wastes (white steel slag, WSS; and furnace iron slag, FIS). The formulations included: (T0) exclusively mine Technosol (control); (T1) 60 % WCS + 40 % WSS; (T2) 60 % WCS + 40 % FIS; and (T3) 50 % WCS + 16.66 % WSS + 33.33 % FIS. The analyses covered leachate quality, soil pore water chemistry, and plant response (germination and survival rates, plant height, and leaf number). Results revealed a significant reduction in leachable contaminant concentrations, with Pb (26.16 mg kg-1), Zn (4.94 mg kg-1), and Cu (2.29 mg kg-1) dropping to negligible levels and shifting towards less toxic species. These changes improved soil conditions, promoting seed germination and seedling growth. Among the formulations tested, Technosol T1 showed promise in overcoming mine soil limitations, enhancing plant adaptation, buffering against acidification, and stabilizing contaminants through precipitation and adsorption mechanisms. The paper stresses the importance of tailoring waste amendments to specific soil conditions, and highlights the broader implications of the Technosol approach, such as waste valorization, soil stabilization, and insights for Brassica juncea growth in extreme environments, including Martian soil simulants.

A Comprehensive Study on Tear Meniscus Height Inter-Eye Differences in Aqueous Deficient Dry Eye Diagnosis.

(1) Background: Dry eye disease (DED) is a chronic ocular surface condition that requires precise diagnostic tools. The present study aimed to investigate the diagnostic potential of the absolute inter-eye difference (|OD-OS|) in tear meniscus height (TMH) for the detection of the presence of aqueous deficient dry eye (ADDE). (2) Methods: A sample of 260 participants with dry eye complaints underwent ocular surface examinations thorough diagnostic assessments based on the Tear Film and Ocular Surface Society guidelines (TFOS DEWS II). Participants were subsequently categorized as No ADDE and ADDE based on TMH. Statistical analyses to determine the optimal TMH|OD-OS| cut-off value in a randomly selected study group (200 participants) were performed, while a separate validation analysis of the cut-off value obtained in a random cross-validation group (60 participants) was also performed. (3) Results: The significant diagnostic capability of TMH|OD-OS| (area under the curve = 0.719 ± 0.036, p < 0.001) was found. The identified cut-off value of 0.033 mm demonstrated reliable specificity (77.6%) and moderate sensitivity (59.1%). Cross-validation confirmed the cut-off value's association with the TFOS DEWS II diagnostic criterion (Cramer's V = 0.354, p = 0.006). (4) Conclusions: The present study provides evidence for the diagnostic potential of TMH|OD-OS| in identifying ADDE. The identified cut-off value enhances the specificity and offers moderate sensitivity, providing an objective tool for clinical decision making.

ctDNA transiting into urine is ultrashort and facilitates noninvasive liquid biopsy of HPV+ oropharyngeal cancer.

BACKGROUNDTransrenal cell-free tumor DNA (TR-ctDNA), which transits from the bloodstream into urine, has the potential to enable noninvasive cancer detection for a wide variety of nonurologic cancer types.MethodsUsing whole-genome sequencing, we discovered that urine TR-ctDNA fragments across multiple cancer types are predominantly ultrashort (<50 bp) and, therefore, likely to be missed by conventional ctDNA assays. We developed an ultrashort droplet digital PCR assay to detect TR-ctDNA originating from HPV-associated oropharyngeal squamous cell carcinoma (HPV+ OPSCC) and confirmed that assaying ultrashort DNA is critical for sensitive cancer detection from urine samples.ResultsTR-ctDNA was concordant with plasma ctDNA for cancer detection in patients with HPV+ OPSCC. As proof of concept for using urine TR-ctDNA for posttreatment surveillance, in a small longitudinal case series, TR-ctDNA showed promise for noninvasive detection of recurrence of HPV+ OPSCC.ConclusionOur data indicate that focusing on ultrashort fragments of TR-ctDNA will be important for realizing the full potential of urine-based cancer diagnostics. This has implications for urine-based detection of a wide variety of cancer types and for facilitating access to care through at-home specimen collections.FundingNIH grants R33 CA229023, R21 CA225493; NIH/National Cancer Institute grants U01 CA183848, R01 CA184153, and P30CA046592; American Cancer Society RSG-18-062-01-TBG; American Cancer Society Mission Boost grant MBGI-22-056-01-MBG; and the A. Alfred Taubman Medical Research Institute.

Gene-expression signature of tumor recurrence in patients with stage II and III colon cancer treated with 5'fluoruracil-based adjuvant chemotherapy.

Although receiving adjuvant chemotherapy after radical surgery, a disappointing proportion of patients with colorectal cancer will develop tumor recurrence. Probability of relapse is currently predicted from pathological staging, there being a need for additional markers to further select high-risk patients. This study was aimed to identify a gene-expression signature to predict tumor recurrence in patients with Stages II and III colon cancer treated with 5'fluoruracil (5FU)-based adjuvant chemotherapy. Two-hundred and twenty-eight patients diagnosed with Stages II-III colon cancer and treated with surgical resection and 5FU-based adjuvant chemotherapy were included. RNA was extracted from formalin-fixed, paraffin-embedded tissue samples and expression of 27 selected candidate genes was analyzed by RT-qPCR. A tumor recurrence predicting model, including clinico-pathological variables and gene-expression profiling, was developed by Cox regression analysis and validated by bootstrapping. The regression analysis identified tumor stage and S100A2 and S100A10 gene expression as independently associated with tumor recurrence. The risk score derived from this model was able to discriminate two groups with a highly significant different probability of tumor recurrence (HR, 2.75; 95%CI, 1.71-4.39; p = 0.0001), which it was maintained when patients were stratified according to tumor stage. The algorithm was also able to distinguish two groups with different overall survival (HR, 2.68; 95%CI, 1.12-6.42; p = 0.03). Identification of a new gene-expression signature associated with a high probability of tumor recurrence in patients with Stages II and III colon cancer receiving adjuvant 5FU-based chemotherapy, and its combination in a robust, easy-to-use and reliable algorithm may contribute to tailor treatment and surveillance strategies.