Immunity of the ocular surface: gene expression of cells collected from the healthy ocular surface
Bibliographic record
Abstract
Purpose The ocular surface varies during the day. Several studies have reported the diurnal changes in the ocular surface epithelial mitosis rate, shedding rate, the concentration of some tear film proteins and ocular surface physiology. However, changes in gene expression that occur in conjunction with these daily changes have not been previously investigated. This study aimed to determine if there is diurnal variation in gene expression in normal healthy conjunctival cells. Methods Two swabs (morning and evening) were collected on the same day from the bulbar conjunctiva of four healthy participants. The morning and evening swab samples were collected from one eye of each subject, with five hours or more gap between the two samples. RNA was isolated with the Single Cell RNA Purification Kit (Norgen Biotek, Thorold, Ontario, Canada) and analysed using RNA sequencing (RNA-Seq). Additionally, the expression patterns of three upregulated genes in the morning and three upregulated genes in the evening were validated in an independent population of six participants using RT-PCR. Results In total, 121 genes were differentially expressed and identified between the morning and evening samples. There were 94 upregulated genes in the morning and 27 upregulated genes in the evening. Many upregulated genes in the morning were associated with gene expression regulation, cell turnover, ocular surface defence, and limbal stem cell function. In contrast, the upregulated genes in the evening were involved in mucin production. Six genes were selected for RT-PCR confirmation of their changes in expression using the RNA seq and this demonstrated that CIART was the only gene confirmed to be significantly upregulated in the morning. Conclusions This study has found that the conjunctival gene expression changes during the day. The findings from this pilot study need to be confirmed with a larger sample size study. Knowledge of the daily rhythms of conjunctival gene expression highlights the regulatory mechanisms of the healthy ocular surface and provides a baseline for studying variations during eye disease.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.002 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".