Assessment of Tear Film Parameters in Smokers and Subjects with a High Body Mass Index.

SIGNIFICANCE: The current study compares the ocular tear film parameters in three different groups using a single noninvasive, practical, and easy-to-use tool. PURPOSE: This study aimed to assess the tear film in smokers, those with a high body mass index (BMI), and healthy subjects using the EASYTEAR view+. METHODS: Thirty men with a high BMI (>25 kg/m2; 24.4 ± 6.4 years), 30 smokers (25.1 ± 6.1 years), and 30 healthy subjects (22.2 ± 3.5 years) were recruited. Each subject completed the Ocular Surface Disease Index, followed by the assessment of noninvasive tear breakup time, tear meniscus height (TMH), and lipid layer patterns (LLPs). RESULTS: Significant differences were found in the median TMH scores between smokers and healthy subjects (P = .03) and between subjects with a high BMI and the healthy ones (P = .04). The median LLP score was significantly (P < .001) higher in normal subjects (4.0 [1.0]) than in smokers (2.4 [1.0]) and subjects with high BMI (2.0 [1.3]). For subjects with a high BMI, the noninvasive tear breakup time score was strongly correlated (Spearman rank correlation coefficient; r) with TMH (r = 0.552, P = .002) and LLP (r = 0.555, P = .001). The LLP showed that grade B (lipid layer thickness, 30 to 50 nm; more compact) was common in subjects with a high BMI, grade C (50 to 80 nm, gray waves) was predominant in smokers, and grade D (~80 nm, dense white-blue layer) represented the majority of normal eye subjects. CONCLUSIONS: Smokers and individuals with a high BMI showed significantly lower lipid layer grades and tear meniscus height scores compared with the control group. The assessment of tear film parameters using the EASYTEAR view+ supports the findings of previous studies that implicate smoking and high BMI as risk factors for dry eye.

Inhibitory Effect of Ursolic Acid on Ultraviolet B Radiation-Induced Oxidative Stress and Proinflammatory Response-Mediated Senescence in Human Skin Dermal Fibroblasts.

Ultraviolet radiation is an environmental carcinogenic agent that enhances inflammation and immunological reactions in the exposed human skin cells leading to oxidative photoaging of the epidermal and dermal segment. In the present study, we investigated the protective role of ursolic acid (UA) against ultraviolet B (UVB) radiation- induced photoaging an in vitro model of human skin dermal fibroblasts. UA-pretreated human skin dermal fibroblast (HDF) cells were exposed to UVB radiation to evaluated cell viability, reactive oxygen species (ROS), mitochondrial membrane potential, lipid peroxidation, antioxidant status, DNA damage, proinflammatory response, apoptotic induction, and matrix metalloproteinase (MMP) alteration. The UA pretreatment of HDFs mitigated the UVB irradiation-induced cytotoxicity, ROS generation, and mitochondrial membrane potential alteration and lipid peroxidation, depletion of antioxidant status, DNA damage, and apoptotic induction. UA pretreatment of HDFs also attenuated the UVB-induced expression of inflammatory (TNF-α and NF-κB) and apoptotic (p53, Bax, and caspase-3) and MMPs (MMP-2 and MMP-9) and enhanced the Bcl-2 protein levels in 20 µM UA treatment, when compared to concentrations. Hence, these results revealed that UA has the potential to mitigate UVB-induced extracellular damage by interfering with the ROS-mediated apoptotic induction and photoaging senescence and thus is a potential therapeutic agent to protect the skin against UVB-irradiation induced photooxidative damage.

Clinical and Ultrastructural Studies of Gelatinous Drop-Like Corneal Dystrophy (GDLD) of a Patient with TACSTD2 Gene Mutation.

PURPOSE: To describe clinical, molecular genetics, histopathologic and ultrastructural findings of gelatinous drop-like corneal dystrophy (GDLD) (OMIM #204870) in a Sudanese patient. METHOD: An ocular examination revealed the onset of GDLD in a Sudanese patient (50 years old) at King Khalid Specialist Hospital, Riyadh. The 333 sequence variants in 13 GDLD genes of a DNA sample were screened by Asper Ophthalmics Ltd. It was further confirmed by sequencing. The patient had undergone a penetrating keratoplasty in the right eye. The corneal tissue was processed for histopathology and ultrastructural studies. RESULTS: Slit-lamp observation showed grayish-white multiple superficial corneal nodules of various sizes in the left and right eye. Both corneas became clear after the surgery. The GDLD deposits in the subepithelial region and in the anterior stroma were confirmed by PAS staining and their apple-green birefringence under polarized light. Ultrastructurally, the amyloid fibrils were very thin and grouped in whorl-like structures, which caused splits between and within the stromal lamellae. Collagen fibrils (CFs) and keratocytes had degenerated. A homozygous c.355T > A mutation in exon 1 of the TACSTD2 (M1S1) gene was detected, and alteration of the amino acid (p.Cysl19Ser in NCBI entry NP_002344.2) was observed. CONCLUSION: In our patient with GDLD, a "c.355T > A" mutation in exon 1 of TACSTD2 was detected and believed to be responsible for the alteration of the amino acid leading to the formation of the amyloid deposits. The deposits caused the ultrastructural degeneration of epithelium, Bowman's layer, stroma, and keratocytes of the GDLD cornea.

Screening and structural evaluation of deleterious Non-Synonymous SNPs of ePHA2 gene involved in susceptibility to cataract formation.

Age-related cataract is clinically and genetically heterogeneous disorder affecting the ocular lens, and the leading cause of vision loss and blindness worldwide. Here we screened nonsynonymous single nucleotide polymorphisms (nsSNPs) of a novel gene, EPHA2 responsible for age related cataracts. The SNPs were retrieved from dbSNP. Using I-Mutant, protein stability change was calculated. The potentially functional nsSNPs and their effect on protein was predicted by PolyPhen and SIFT respectively. FASTSNP was used for functional analysis and estimation of risk score. The functional impact on the EPHA2 protein was evaluated by using SWISSPDB viewer and NOMAD-Ref server. Our analysis revealed 16 SNPs as nonsynonymous out of which 6 nsSNPs, namely rs11543934, rs2291806, rs1058371, rs1058370, rs79100278 and rs113882203 were found to be least stable by I-Mutant 2.0 with DDG value of > -1.0. nsSNPs, namely rs35903225, rs2291806, rs1058372, rs1058370, rs79100278 and rs113882203 showed a highly deleterious tolerance index score of 0.00 by SIFT server. Four nsSNPs namely rs11543934, rs2291806, rs1058370 and rs113882203 were found to be probably damaging with PSIC score of ≥ 2. 0 by Polyp hen server. Three nsSNPs namely, rs11543934, rs2291806 and rs1058370 were found to be highly polymorphic with a risk score of 3-4 with a possible effect of Non-conservative change and splicing regulation by FASTSNP. The total energy and RMSD value was higher for the mutant-type structure compared to the native type structure. We concluded that the nsSNP namely rs2291806 as the potential functional polymorphic that is likely to have functional impact on the EPHA2 gene.

Role of epidermal growth factor receptor (EGFR) in corneal remodelling in diabetes.

PURPOSE: This study examined the role of epidermal growth factor receptor (EGFR) signalling on the organization and remodelling of collagen fibrils (CFs) and proteoglycans (PGs) in the stroma of diabetic rat cornea. METHODS: Diabetes was induced in female Wistar rats (n = 5) by streptozotocin (STZ) injection (55 mg/kg). Treatment with a selective inhibitor of EGFR tyrosine kinase, AG1478, was started on the same day as the induction of diabetes and administered every other day for 4 weeks. Corneas were fixed in 4% paraformaldehyde at 4 degrees to allow for analysis of CF diameters and in 2.5% glutaraldehyde in sodium acetate buffer containing cuprolinic blue to enable the study of PG distribution. AnalySIS soft imaging software was used to analyse CFs and PGs. RESULTS: Epithelial thickness, and median diameter and area fraction of CF in corneal stroma were decreased in diabetic rat cornea compared with normal cornea (p < 0.001), whereas the median PG area and area fractions were significantly increased (p < 0.001). Treatment with AG1478, although it had no action on normal cornea, prevented these diameter and area fraction changes in CFs and PGs. The cornea of AG1478-treated diabetic rats showed a slight increase in CF diameter and area fraction and a decreased number density. CONCLUSIONS: These data show that the distribution of corneal stroma CFs and PGs was altered after 4 weeks of diabetes and that, furthermore, treatment with an EGFR signalling inhibitor normalized these abnormalities. The data suggest that EGFR plays an important role in the development of diabetes-induced corneal remodelling.

Repeatability of central corneal thickness measurements measured with the Topcon SP2000P specular microscope.

BACKGROUND: The non-contact specular microscope has become the method of choice for a quick, accurate and non-invasive assessment of central corneal thickness (CCT), which is an important variable to monitor before and after refractive surgery. The consistency of the results produced by such widely used methods/equipment must be assessed to determine their reliability. The purpose of this study was to assess within- and between-observer repeatability of, and to determine if a systematic bias exists in the measurements made by, the Topcon SP2000P specular microscope. METHODS: The CCT of the right eyes of 70 adult subjects, divided equally between men and women, was assessed on two separate occasions (4-7 days apart) by each of two examiners using the low-intensity auto mode of the SP2000P specular microscope. RESULTS: The average CCT values for men and women, measured by one observer, were 0.52+/-0.03 mm (mean +/- SD) and 0.52+/-0.04 mm, respectively. The average for the entire sample was 0.52+/-0.04 mm. Within- and between-observer repeatability were assessed by plotting the mean difference (for each subject) between two readings made by the same observer or one each by both observers against the combined average CCT reading of both sessions; the mean difference between two sets of observations was not significantly different from zero (P<0.05). For the first observer, the 95% limits of repeatability were between -0.015 and 0.017 mm. For the second observer, the 95% limits of repeatability were between -0.018 and 0.018 mm. For the between-observer repeatability, the 95% limits of agreement were between -0.016 and 0.016 mm. For both within- and between-observer repeatability, we found no systematic bias of the mean difference with the average CCT reading. CONCLUSION: The within- and between-observer limits of agreement we found were similar to those previously reported for the Topcon SP2000P specular microscope; the range of the 95% limits of repeatability were within +/-1 SD of the average CCT reading for both sessions. We suggest that a technique be considered reliable if: (1) the mean difference between two measurements does not vary significantly from zero, (2) there is no systematic bias of the mean difference with the magnitude of the measured quantity and (3) the error inherent in a measurement technique is within +/-1 SD of the average measurement of the two sessions.