Serum metabolomics profiling and potential biomarkers of myopia using LC-QTOF/MS.

Myopia is the most common form of refractive eye disease, and the prevalence is increasing rapidly worldwide. However, the key metabolic alterations in individuals with high myopia are not understood clearly, and serum biomarkers remain to be determined. The objectives of this study were to identify serum biomarkers and investigate the metabolic alterations of myopia. The serum metabolomics profiling was investigated on 30 high myopia cases and 30 controls (without myopia) using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), and an independent additional cohort including 20 cases and 19 controls were investigated to validate potential metabolite candidates for biomarkers. According to the metabolic differences, the myopia patients and controls could be divided into different clusters and nine metabolites were found to be closely correlated with myopia. In the cohort of validation, eight metabolites were confirmed. Metabolic pathway analyses of these metabolites of high myopia involved abnormal phospholipid, diacylglycerol, amino acid, and vitamin metabolism, which were closely correlated with oxidative stress and inflammation. Multiple logistic regression analyses showed that γ-glutamyltyrosine and 12-oxo-20-trihydroxy-leukotriene B4 were potential biomarkers of myopia with a combined high sensitivity (97%), specificity (90%), and area under the curve value (0.983). These findings may contribute to an understanding of the pathophysiological changes and pathogenesis of myopia, and provide novel insight into the early prevention and control of high myopia.

Time outdoors, blood vitamin D status and myopia: a review.

BACKGROUND: Myopia is a major public health concern throughout the world and the prevalence has been increasing rapidly in recent years, especially in urban Asia. The "vitamin D hypothesis" has been raised recently because vitamin D may be a link between less time outdoors and increased risk of myopia. METHODS: We reviewed all studies published in English which examined the association of time outdoors and blood vitamin D status with myopia. RESULTS: The protective effect of time spent outdoors on the risk of myopia onset has been well-established with numerous observational studies and three trials published. Five studies reporting the association between the blood vitamin D status and the risk of myopia and two studies examining the variations in the vitamin D receptor as potential risk factors for myopia development were identified. Most of the current evidence was cross-sectional in nature and had not properly controlled important confounders in its analyses. The evidence supporting that vitamin D played a role in myopia development is weak and the mechanisms are unclear. CONCLUSIONS: At the current stage, it is still unclear whether blood vitamin D status regulates the onset or progression of myopia. Blood vitamin D status may only serve as a biomarker of outdoor exposure, which is the real protective factor for myopia.

Myopes have significantly higher serum melatonin concentrations than non-myopes.

PURPOSE: Experimental animal models of myopia demonstrate that higher melatonin (Mel) and lower dopamine (DA) concentrations actively promote axial elongation. This study explored the association between myopia and serum concentrations of DA and Mel in humans. METHODS: Morning serum concentrations of DA and Mel were measured by solid phase extraction-liquid chromatography-tandem mass spectrometry from 54 participants (age 19.1 ± 0.81 years) in September/October 2014 (phase 1) and March/April 2016 (phase 2). Axial length (AL), corneal radii (CR) and spherical equivalent refraction (SER) were also recorded. Participants were defined as myopic if non-cycloplegic spherical equivalent refractive error ≤-0.50 DS at phase 1. RESULTS: Nine participants were lost to follow up. Mel concentrations were measurable for all myopes (phase 1 n = 25, phase 2 n = 22) and non-myopes (phase 1 n = 29, phase 2 n = 23). SER did not change significantly between phases (p = 0.51). DA concentrations were measurable for fewer myopes (phase 1 n = 13, phase 2 n = 12) and non-myopes (phase 1 n = 23, phase 2 n = 16). Myopes exhibited significantly higher Mel concentrations than non-myopes at phase 1 (Median difference: 10 pg mL-1 , p < 0.001) and at phase 2 (Median difference: 7.3 pg mL-1 , p < 0.001) and lower DA concentrations at phase 2 (Median difference: 4.7 pg mL-1 , p = 0.006). Mel concentrations were positively associated with more negative SER (all r ≥ -0.53, all p < 0.001), longer AL (all r ≥ 0.37, all p ≤ 0.008) and higher AL/CR ratio (all r ≥ 0.51, all p < 0.001). CONCLUSION: This study reports for the first time in humans that myopes exhibit higher serum Mel concentrations than non-myopes. This may indicate a role for light exposure and circadian rhythm in the human myopic growth mechanism. Further research should focus on younger cohorts exhibiting more dynamic myopic progression and explore the profile of these neurochemicals alongside evaluation of sleep patterns in myopic and non-myopic groups.

Rheumatic Disease Autoantibodies in Autoimmune Liver Diseases.

BACKGROUND: Autoimmune liver diseases (ALDs) are known to be associated with systemic autoimmune rheumatic diseases (SARDs) and their autoantibodies. We aimed to study the prevalence of SARDs and related autoantibodies, as well as their prognostic implications in a group of patients with ALDs. METHODS: This was a cross-sectional study. Sixty patients with ALDs (38.3% with autoimmune hepatitis; 11.7% with primary biliary cirrhosis; 25% with primary sclerosing cholangitis and 25% with overlap syndrome) were studied for the presence of SARDs and their autoantibodies. RESULTS: There was autoimmune rheumatic disease in 20% of the studied sample. Systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) were the commonest (11.6% and 5%, respectively). Antinuclear antibodies (ANAs) were present in 35% of the patients, followed by anti-Ro (20.0%); anti-nucleosome (18.3%); rheumatoid factor (10%) anti-CCP (8.3%); anti-RNP (8.3%); anti-ds-DNA (6.6%); anti-La (3.3%); anti-Sm (3.3%), anti-ribosomal P (3.3%). Anti-Ro (p = 0.0004), anti-La (p = 0.03), anti-RNP (p = 0.04) and anti-Sm (p = 0.03) were commonly found in patients with SARD, but not anti-DNA, anti-nucleosome and anti-ribosomal P. No differences were found in liver function tests regarding to the presence of autoantibodies. CONCLUSIONS: There was a high prevalence of SARD and their autoantibodies in ALD patients. Anti-Ro, anti-La, anti-RNP and anti-Sm positivity points to an association with systemic autoimmune rheumatic diseases. The presence of autoantibodies was not related to liver function tests.

Low serum vitamin D is associated with axial length and risk of myopia in young children.

The aim of the study was to investigate the relationship between serum 25(OH)D levels and axial length (AL) and myopia in 6-year-old children. A total of 2666 children aged 6 years participating in the birth-cohort study Generation R underwent a stepwise eye examination. First, presenting visual acuity (VA) and AL were performed. Second, automated cycloplegic refraction was measured if LogMAR VA > 0.1. Serum 25-hydroxyvitamin D [25(OH)D] was determined from blood using liquid chromatography/tandem mass spectrometry. Vitamin D related SNPs were determined with a SNP array; outdoor exposure was assessed by questionnaire. The relationships between 25(OH)D and AL or myopia were investigated using linear and logistic regression analysis. Average 25(OH)D concentration was 68.8 nmol/L (SD ± 27.5; range 4-211); average AL 22.35 mm (SD ± 0.7; range 19.2-25.3); and prevalence of myopia 2.3 % (n = 62). After adjustment for covariates, 25(OH)D concentration (per 25 nmol/L) was inversely associated with AL (ß -0.043; P < 0.01), and after additional adjusting for time spent outdoors (ß -0.038; P < 0.01). Associations were not different between European and non-European children (ß -0.037 and ß -0.039 respectively). Risk of myopia (per 25 nmol/L) was OR 0.65 (95 % CI 0.46-0.92). None of the 25(OH)D related SNPs showed an association with AL or myopia. Lower 25(OH)D concentration in serum was associated with longer AL and a higher risk of myopia in these young children. This effect appeared independent of outdoor exposure and may suggest a more direct role for 25(OH)D in myopia pathogenesis.

[Analysis on the relationship between adolescent myopia and serum sex hormone].

OBJECTIVE: To investigate the quantitative changes of the serum sex hormone levels in juvenile myopia patients. METHODS: In January 2013, investigation of Wenzhou middle school 822 adolescents, including 432 male patients and 390 female patients were involved in the study Visual acuity ≥ 5 was set as the normal value. The subjects were divided into two groups, the myopia group and the non-myopia group. Chemiluminescence immunoassay analyzer was used to detect the subjects' sex hormone concentration of estradiol (E2), testosterone (T), follicle stimulating hormone (FSH) and luteinizing hormone (LH). RESULTS: The follicle stimulating hormone and luteinizing hormone of the female patients were significantly higher than those of male counterparts (P < 0.01); among the two groups consisting female myopia and non-myopia patients, a significant difference in luteinizing hormone and follicle stimulating hormone was shown (P < 0.05); the distinct differences in the level of luteinizing hormone and testosterone level showed in the male myopia group and non-myopia group were of outstanding statistics significance (P < 0.01), showing the result that the level of sex hormone in myopia group was higher than that in the non-myopia group. CONCLUSION: A close relationship perhaps between the level of sex hormone with gender differences and juvenile myopia is confirmed.

Reduced retinol (vitamin A) and α-tocopherol (vitamin E) blood levels and increased myeloperoxidase (MPO) activity in children with high myopia.

The study assessed selected parameters of redox status in the plasma of patients suffering from high myopia (HM). Thirty-five children with mean age 13.7 ± 2.7 years with HM and 40 healthy children were included. Plasma redox status parameters were determined using colorimetric kits. The levels of retinol, α-tocopherol and coenzyme Q10 were determined with a high-performance liquid chromatograph. Negative correlations were observed between the concentrations of retinol and the axial length of the eye (r = - 0.514 p < 0.001). Increased myeloperoxidase (MPO) activity (p < 0.018), and decreased concentrations of retinol (p < 0.001) and α-tocopherol (p < 0.023) in patients with HM and the axial length of the eye > 26 mm compared to controls were established. Significantly lower retinol and α-tocopherol concentrations were found in patients with the axial length of the eye > 26 mm compared to those with the axial length of the eye ≤ 26 mm (p < 0.001, p < 0.021, respectively). Increased MPO activity in advanced stages of HM may confirm an inflammatory process in HM patients. Reduced retinol and α-tocopherol concentrations and their link to disease progression indicate a need for monitoring their levels and supplementation in children with HM.

Polymorphisms in steroidogenesis genes, sex steroid levels, and high myopia in the Taiwanese population.

PURPOSE: To evaluate the relationship among single nucleotide polymorphisms (SNPs) in steroidogenesis enzyme genes, serum levels of sex steroids, and high myopia in Taiwanese male and female populations. METHODS: A campus-based sample of 283 cases (145 males and 138 females) with high myopia and 280 controls (144 males and 136 females) with low myopia or emmetropia was studied. Estradiol, progesterone, and testosterone levels were determined using enzyme-linked immunosorbent assay kits. We genotyped six SNPs within five steroidogenesis enzyme genes (17 alpha-hydroxylase/17,20 lyase [CYP17A1], 3 beta-hydroxysteroid dehydrogenase [HSD3B1], 17 beta-hydroxysteroid dehydrogenase 1 [HSD17B1], steroid-5-alpha-reductase, alpha polypeptide 2 [SRD5A2], and aromatase [CYP19A1]) using polymerase chain reaction-restriction fragment length polymorphism methods. Student's t-tests, χ(2) tests, logistic regression, multifactor dimensionality reduction (MDR) methods, and ANOVA were used to determine significance. RESULTS: An MDR analysis corroborated the synergistic genotype association and demonstrated that synergistic interaction between rs6203 (HSD3B1), rs10046 (CYP19A1), and sex might confer susceptibility to high myopia (p=0.019). In both male and female subjects, levels of testosterone were significantly higher in cases than in controls; in male subjects, the levels of estradiol were significantly higher and those of progesterone were significantly lower in cases (all p-values <0.001). The rs605059 (HSD17B1), with sex-gene interaction, showed association with estradiol levels in males (p=0.035) and testosterone levels in females (p=0.027). CONCLUSIONS: Testosterone levels correlate with high myopia, and interaction of steroidogenesis enzyme genes and sex may be a modulating factor in sex hormone metabolism and high-myopia risk.

Blood levels of vitamin D in teens and young adults with myopia.

PURPOSE: Longitudinal data suggest that time outdoors may be protective against myopia onset. We evaluated the hypothesis that time outdoors might create differences in circulating levels of vitamin D between myopes and non-myopes. METHODS: Subjects provided 200 µl of peripheral blood in addition to survey information about dietary intakes and time spent in indoor or outdoor activity. The 22 subjects ranged in age from 13 to 25 years. Myopes (n = 14) were defined as having at least -0.75 diopter of myopia in each principal meridian and non-myopes (n = 8) had +0.25 diopter or more hyperopia in each principal meridian. Blood level of vitamin D was measured using liquid chromatography/mass spectroscopy. RESULTS: Unadjusted blood levels of vitamin D were not significantly different between myopes (13.95 ± 3.75 ng/ml) and non-myopes (16.02 ± 5.11 ng/ml, p = 0.29) nor were the hours spent outdoors (myopes = 12.9 ± 7.8 h; non-myopes = 13.6 ± 5.8 h; p = 0.83). In a multiple regression model, total sugar and folate from food were negatively associated with blood vitamin D, whereas theobromine and calcium were positively associated with blood vitamin D. Myopes had lower levels of blood vitamin D by an average of 3.4 ng/ml compared with non-myopes when adjusted for age and dietary intakes (p = 0.005 for refractive error group, model R = 0.76). Gender, time outdoors, and dietary intake of vitamin D were not significant in this model. CONCLUSIONS: The hypothesis that time outdoors might create differences in vitamin D could not be evaluated fully because time outdoors was not significantly related to myopia in this small sample. However, adjusted for differences in the intake of dietary variables, myopes appear to have lower average blood levels of vitamin D than non-myopes. Although consistent with the hypothesis above, replication in a larger sample is needed.

Myopia, Melatonin and Conjunctival Ultraviolet Autofluorescence: A Comparative Cross-sectional Study in Indian Myopes.

Purpose: To explore the role of outdoor light exposure by estimating ocular sun exposure measured by Conjunctival Ultraviolet Autofluorescence (CUVAF) imaging and serum melatonin levels in myopes and non-myopes.Materials & Methods: Age and sex matched emmetropes and myopes (60 each) aged 10-25 years participated. Those with a history of ocular surgery or any ocular or systemic co-morbidity were excluded. Socio-demographic parameters, sun exposure questionnaires, indoor and outdoor activity profile, morning serum melatonin levels, sleep pattern, degree of myopia, ocular biometry and area of CUVAF on ultraviolet photography were noted and analyzed.Results: Mean age of myopes (18 ± 4.5 years) and emmetropes (18.5 ± 4 years) was similar (P = .523). Serum melatonin levels were significantly higher (P = .001) among myopes (89.45 pg/ml) as compared to emmetropes (52.83 pg/ml). Lifetime sun exposure was significantly lower in myopes than emmetropes (P = .0003). Area of CUVAF was inversely related to degree of myopia (P < .0001). Day time sleepiness was greater in myopes (51.7%) than emmetropes (15%) (P < .0001). There was a positive correlation between serum melatonin levels and axial length among myopes (correlation coefficient = 0.27; P = .03). Age and gender had no association with serum melatonin levels.Conclusion: This study demonstrates an inverse relationship between serum melatonin levels and degree of CUVAF in myopes. A novel link between serum melatonin, axial length and outdoor sun exposure is highlighted in the current study.

Myopia in school-aged children with preterm birth: the roles of time spent outdoors and serum vitamin D.

AIMS: To analyse the factors associated with myopia in school-aged children with preterm birth and with or without retinopathy of prematurity (ROP). METHODS: Children born prematurely between January 2010 and December 2011 were enrolled in this cross-sectional study when they reached school age between April 2017 and June 2018 in a referral centre. The main parameters were cycloplegic refraction, time spent outdoors and serum 25-hydroxyvitamin D (25(OH)D) concentration. RESULTS: A total of 99 eyes from 99 children with a mean age of 6.8 years underwent analysis. The average time spent outdoors was significantly higher in the non-myopic group (0.9 ± 0.5 hours/day) than in the myopic group (0.7 ± 0.3 hours/day) (p = 0.032). After adjustment for age, sex, number of myopic parents, ROP severity, near-work time and serum 25(OH)D concentration, more time spent outdoors was correlated with a lower odds of myopia (OR, 0.13 per additional hour per day; 95% CI, 0.02-0.98; p = 0.048). Mean serum 25(OH)D concentrations were similar between the myopic and non-myopic groups (49.7 ± 13.6 and 48.8 ± 14.0 nmol/mL; p = 0.806) and were not correlated with spherical equivalence power (r = -0.09; p = 0.418). Vitamin D insufficiency was present in 57% of the participants. CONCLUSIONS: Among preterm children with or without ROP, more time spent outdoors was associated with lower odds of myopia. The serum 25(OH)D concentration was not associated with myopia, but a high proportion of the participants had insufficient levels.

Time spent outdoors through childhood and adolescence - assessed by 25-hydroxyvitamin D concentration - and risk of myopia at 20 years.

PURPOSE: To investigate the relationship between time spent outdoors, at particular ages in childhood and adolescence, and myopia status in young adulthood using serum 25-hydroxyvitamin D [25(OH)D] concentration as a biomarker of time spent outdoors. METHODS: Participants of the Raine Study Generation 2 cohort had 25(OH)D concentrations measured at the 6-, 14-, 17- and 20-year follow-ups. Participants underwent cycloplegic autorefraction at age 20 years, and myopia was defined as a mean spherical equivalent -0.50 dioptres or more myopic. Logistic regression was used to analyse the association between risk of myopia at age 20 years and age-specific 25(OH)D concentrations. Linear mixed-effects models were used to analyse trajectory of 25(OH)D concentrations from 6 to 20 years. RESULTS: After adjusting for sex, race, parental myopia, body mass index and studying status, myopia at 20 years was associated with lower 25(OH)D concentration at 20 years (per 10 nmol/L decrease, odds ratio (aOR)=1.10, 95% CI: 1.02, 1.18) and a low vitamin D status [25(OH)D < 50 nmol/L] at 17 years (aOR = 1.71, 95% CI: 1.06, 2.76) and 20 years (aOR = 1.71, 95% CI: 1.14, 2.56), compared to those without low vitamin D status. There were no associations between 25(OH)D at younger ages and myopia. Individuals who were myopic at 20 years had a 25(OH)D concentration trajectory that declined, relative to non-myopic peers, with increasing age. Differences in 25(OH)D trajectory between individuals with and without myopia were greater among non-Caucasians compared to Caucasians. CONCLUSIONS: Myopia in young adulthood was most strongly associated with recent 25(OH)D concentrations, a marker of time spent outdoors.

Association study of the serum 25(OH)D concentration and myopia in Chinese children.

ABSTRACT: To analyze the serum 25 hydroxyvitamin D (25[OH]D) concentration in Chinese children with myopia and explore its correlation with myopia.From July to September in 2019, myopic children were collected from the Myopia Influencing Factors Survey Project. The basic information and vision related behaviors of the subjects were collected by questionnaire. The diopter of the children without dilated pupils was measured by the computerized refractometer. Meanwhile, 5 ml fasting venous blood samples were collected for the determination of serum 25(OH)D concentration.A total of 186 children were included in this study, including 90 males and 96 females, with an average age of 8 ±â€Š3.26 years. The detection rate of serum 25(OH)D deficiency in myopic children was 65.59% (122/186). There was statistical significance in the detection rate of serum 25(OH)D deficiency in children with different myopic degrees (χ2 = 6.635, P = .010). The average serum 25(OH)D concentration in myopic children was 14.86 (10.67-18.96) ng/ml, and the difference of serum 25(OH)D concentration in children with different myopia degrees was statistically significant (Z = 20.23, P < .001). Logistic regression analysis showed that after controlling for gender, parental myopia, after-school class, and outdoor activities, the prevalence of developing moderate and high myopia was 2.051 times (95% confidence interval: 1.272-3.724) higher in the serum 25(OH)D deficiency group than in the serum 25(OH)D sufficiency group. There is a positive correlation between serum 25(OH)D concentration and the equivalent spherical degree of myopic children.The study found that serum 25(OH)D concentration is closely related to the prevalence of myopia in Chinese children. The results further support the conclusion that children with a higher level of serum 25(OH)D have a lower prevalence of moderate to high myopia. The results of this study provide a basis for further research into the relationship between vitamin D and visual development in children and its mechanisms.

Higher HbA1c may reduce axial length elongation in myopic children: a comparison cohort study.

AIMS: To compare the annual axial length (AL) changes in myopic children with type 1 diabetes mellitus (T1DM) and those without diabetes. METHODS: There are two groups of myopic children in this retrospective cohort study. Group 1 consisted of myopic children with T1DM (44 eyes of 22 patients). Group 2 comprised age-matched myopic children without diabetes (44 eyes of 22 children). These two groups were compared with regard to their baseline clinical characteristics. A generalized estimating equations (GEE) model was also used to determine the most likely factor that contributed to the results. RESULTS: The average ages of group 1 and group 2 were 14.8 and 14.6 years, respectively. Children in group 1 had significantly slower annual AL changes (0.051 mm/year vs 0.103 mm/year; 50.5% slower, P = 0.011) and shorter baseline AL (23.97 vs 25.19 mm, P < 0.001) than those in group 2. GEE also showed that serum glycated hemoglobin (HbA1c) level (B = -0.023, P = 0.039) was the most important factor in reducing AL elongation in group 1 myopic children. CONCLUSIONS: Long-term higher HbA1c level may reduce AL elongation. A strict blood sugar control strategy in clinical practice is warranted to axial myopia progression in T1DM children.

No evidence for an association of plasma homocysteine levels and refractive error - Results from the population-based Gutenberg Health Study (GHS).

PURPOSE: There is a strong association between severe hyperhomocysteinemia and myopia. Thus we studied the hypothesis that even moderately increased levels of homocysteine (Hcy) might be a potentially treatable risk factor for myopia. METHODS: The Gutenberg Health Study (GHS) is a population-based, prospective, observational cohort study in Germany, including 15,010 participants aged between 35 and 74 at recruitment. The baseline examination was conducted from 2007-2012. Refraction was measured using autorefraction (HARK 599, Carl Zeiss AG, Jena, Germany). Hcy was measured by an immunoassay. We included only phakic participants without a history of corneal surgery or corneal laser treatment. We used linear regression models to evaluate the potential association between Hcy and refraction at baseline, and between Hcy and change in refraction between baseline and 5-year-follow-up examination. We used generalized estimating equation models to account for the correlation between fellow eyes. RESULTS: We included 13,749 participants, categorized as having no myopia (spherical equivalent > -0.75 D, 65.2%), low myopia (-0.75 D--2.75 D, 21.5%), moderate myopia (-3.00 D- 5.75 D, 9.8%) and high myopia (≤ -6 D, 3.5%). Median Hcy levels were similar in all groups (µmol/l). We observed no association of Hcy with refraction or 5-year change in refraction in the models adjusted for age, sex and socioeconomic status. CONCLUSION: We found no evidence for an association of Hcy levels and refractive error.

Association between serum 25-hydroxyvitamin D levels and myopia in general Korean adults.

Purpose: We performed this study to determine the association between serum 25-hydroxyvitamin D [25(OH) D] level and myopia in adults. Methods: A total of 25,199 subjects aged ≥20 years were included from the National Health and Nutrition Examination Survey 2008-2012. Blood 25(OH)D levels were evaluated from blood samples. Refractive error was measured without cycloplegia. Myopia and high myopia were defined as ≥-0.50 diopters (D) and ≥-6.0 D, respectively. Other covariates such as education, physical activity, and economic status were obtained from interviews. Results: Linear regression analysis showed that as 25(OH) D level increased by 1 ng/mL, myopic refractive error significantly decreased by 0.01 D (P < 0.001) after adjusting for potential confounders including sex, age, height, education level, economic status, physical activity, and sunlight exposure time. The odds ratios for myopia was 0.75 (95% Confidence interval [CI]; 0.67-0.84, P < 0.001) in the highest 25(OH) D quintile compared to the lowest quintile. The odds ratios for high myopia was 0.63 (95% CI; 0.47-0.85, P < 0.001) in the highest 25(OH)D quintile compared to the lowest quintile. Conclusion: : Serum 25(OH)D level was inversely associated with myopia in Korean adults.

Vitamin D and its pathway genes in myopia: systematic review and meta-analysis.

OBJECTIVE: To conduct a systematic review and meta-analysis of the association of blood vitamin D (25-hydroxyvitamin D, 25(OH)D) concentration and vitamin D pathway genes with myopia. METHODS: We searched the MEDLINE and EMBASE databases for studies published up to 29 January 2018. Cross-sectional or cohort studies which evaluated the blood 25(OH)D concentration, blood 25(OH)D3 concentration or vitamin D pathway genes, in relation to risk of myopia or refractive errors were included. Standard mean difference (SMD) of blood 25(OH)D concentrations between the myopia and non-myopia groups was calculated. The associations of blood 25(OH)D concentrations and polymorphisms in vitamin D pathway genes with myopia using summary ORs were evaluated. RESULTS: We summarised seven studies involving 25 008 individuals in the meta-analysis. The myopia group had lower 25(OH)D concentration than the non-myopia group (SMD=-0.27 nmol/L, p=0.001). In the full analysis, the risk of myopia was inversely associated with blood 25(OH)D concentration after adjusting for sunlight exposure or time spent outdoors (adjusted odds ratio (AOR)=0.92 per 10 nmol/L, p<0.0001). However, the association was not statistically significant for the <18 years subgroup (AOR=0.91 per 10 nmol/L, p=0.13) and was significant only for 25(OH)D3 (likely to be mainly sunlight derived), but not total 25(OH)D (AOR=0.93 per 10 nmol/L, p=0.00007; AOR=0.91 per 10 nmol/L, p=0.15). We analysed four single nucleotide polymorphisms in the VDR gene from two studies; there was no significant association with myopia. CONCLUSIONS: Lower 25(OH)D is associated with increased risk of myopia; the lack of a genetic association suggests that 25(OH)D level may be acting as a proxy for time outdoors.

Conjunctival ultraviolet autofluorescence area, but not intensity, is associated with myopia.

BACKGROUND: Conjunctival ultraviolet autofluorescence (CUVAF) has been used as a biomarker of time spent outdoors. Smaller CUVAF area is associated with myopia in southern hemisphere cohorts. Further research is required to determine if this association is replicated in northern latitudes and whether average CUVAF intensity is a valuable metric. This prospective study explored the association between myopia, CUVAF (area and intensity) and additional indicators of sun exposure (vitamin D3 and self-reported sun exposure preferences) across seasons at a location of 55° north. METHODS: Young adults (age 18-20) provided blood samples biannually (March/April and September/October) over an 18-month period (four phases) for the assessment of 25-hydroxyvitamin D (25(OH)D3 ) concentrations (liquid chromatography-tandem mass spectrometry). CUVAF (total area, average intensity) and self-reported sun exposure preferences were recorded at each phase. Axial length and corneal radius were measured. Refractive error was measured by autorefractor and spherical equivalent refraction used to classify participants into refractive groups: myopic (spherical equivalent refraction ≤ -0.50 DS) or non-myopic. RESULTS: Fifty-four participants (24 myopes, 30 non-myopes) participated. CUVAF area was negatively associated with the presence of myopia (odds ratio = 0.94, 95 per cent confidence interval = 0.90-0.98, p = 0.002). Myopes = 4.5 mm2 (interquartile range [IQR] 0.95-6.4 mm2 ), non-myopes = 7.0 mm2 (IQR = 2.0-10.7 mm2 ). No significant association was found between CUVAF intensity and refractive group (p = 0.17). There was no significant association between sun exposure preferences or serum concentration of 25(OH)D3 and refractive status (all p ≥ 0.21). CUVAF measures were not associated with ocular biometry measures (all p ≥ 0.084). CUVAF area was unaffected by season (all p ≥ 0.45) and variations in CUVAF area over the study period did not exceed the repeatability of the measurement technique. CONCLUSION: Myopia was associated with smaller areas of CUVAF indicative of less cumulative ultraviolet-B exposure. These findings suggest that CUVAF measures are a useful, non-invasive biomarker of the time spent outdoors in adults in northern hemisphere populations.

Independent association of whole blood miR-328 expression and polymorphism at 3'UTR of the PAX6 gene with myopia.

PURPOSE: This study aimed to find associations between miR-328 expression in whole blood, polymorphism at 3'UTR of the PAX6 gene (paired box homeotic gene 6) and myopia. METHODS: We evaluated 451 individuals (142 individuals with low, 49 with moderate and 13 with high-degree myopia, and 247 healthy individuals). DNA and RNA were extracted from peripheral blood samples. Expression of miR-328 was assessed and genotyping of single-nucleotide polymorphisms (SNPs) of the PAX6 (rs662702) performed using the Applied Biosystems 7900HT Real-Time Polymerase Chain Reaction System. RESULTS: Moderate and high degree myopia showed significant differences between TT and CT genotypes of the PAX6 gene (p < 0.001). In the myopia group, 71.4% of the subjects had the TT genotype and 28.6% had the CT genotype; meanwhile in the control group, 97.1% had the TT genotype and 2.9% had the CT genotype. The odds ratio of having moderate and/or high degree myopia for individuals with the CT genotype was 13.6 (2.865-64.55) 95% CI versus TT genotype (p = 0.001). MiR-328 results showed that ∆Ct values differed statistically significantly between the myopia and control groups. Patients with myopia in the peripheral blood cells had a higher expression of miR-328 than controls (p < 0.05). CONCLUSIONS: Significant differences were detected between the PAX6 gene (rs662702) TT and CT genotypes in moderate and high degree myopia; the risk C allele increased the risk for myopia. The expression level of miR-328 in peripheral blood cells was higher in patients with myopia than controls. We did not find the association between expression of mir-328 in the peripheral blood cells and PAX6 gene (rs662702) polymorphism comparing myopia and control groups.

Genetically low vitamin D concentrations and myopic refractive error: a Mendelian randomization study.

Background: Myopia prevalence has increased in the past 20 years, with many studies linking the increase to reduced time spent outdoors. A number of recent observational studies have shown an inverse association between vitamin D [25(OH)D] serum levels and myopia. However, in such studies it is difficult to separate the effects of time outdoors and vitamin D levels. In this work we use Mendelian randomization (MR) to assess if genetically determined 25(OH)D levels contribute to the degree of myopia. Methods: We performed MR using results from a meta-analysis of refractive error (RE) genome-wide association study (GWAS) that included 37 382 and 8 376 adult participants of European and Asian ancestry, respectively, published by the Consortium for Refractive Error And Myopia (CREAM). We used single nucleotide polymorphisms (SNPs) in the DHCR7, CYP2R1, GC and CYP24A1 genes with known effects on 25(OH)D concentration as instrumental variables (IV). We estimated the effect of 25(OH)D on myopia level using a Wald-type ratio estimator based on the effect estimates from the CREAM GWAS. Results: Using the combined effect attributed to the four SNPs, the estimate for the effect of 25(OH)D on refractive error was -0.02 [95% confidence interval (CI) -0.09, 0.04] dioptres (D) per 10 nmol/l increase in 25(OH)D concentration in Caucasians and 0.01 (95% CI -0.17, 0.19) D per 10 nmol/l increase in Asians. Conclusions: The tight confidence intervals on our estimates suggest the true contribution of vitamin D levels to degree of myopia is very small and indistinguishable from zero. Previous findings from observational studies linking vitamin D levels to myopia were likely attributable to the effects of confounding by time spent outdoors.