Opposite Cell Density-Dependence between Spontaneous and Oxidative Stress-Induced Apoptosis in Mouse Fibroblast L-Cells
Bibliographic record
Abstract
BACKGROUND/AIMS: Spontaneous apoptosis in culture is cell density-dependent. Yet, little is known about the density-dependence of apoptosis induced by oxidative stress and the underlying signaling mechanisms. METHODS: We compared apoptosis occurring spontaneously and induced by H2O2 in mouse fibroblast L-cells seeded at three different densities, using ELISA and TUNEL to measure DNA fragmentation, Western blot and pharmacological probes to evaluate the roles of signaling molecules. RESULTS: The high-density (HD, 2.5 x 10(5)/cm2) culture had approximately 1.5- and approximately 3-times more spontaneous apoptosis than medium-density (MD, 5 x 10(4)/cm2) and low-density (LD, 1 x 10(4)/cm2) cultures, respectively. The opposite was seen with H2O2 (500 microM)-induced apoptosis: more apoptosis in LD than in MD and HD. Caspase inhibitor (Z-VAD-fmK) diminished the density-dependence of both spontaneous and H2O2-induced apoptosis. Activation of caspase 3 by H2O2 was more pronounced in LD than in MD and HD. Inhibition of PI3K/Akt increased spontaneous apoptosis more in LD and H2O2-induced apoptosis more in HD, diminishing the density-dependence of apoptosis. p38 MAPK inhibition reduced H2O2-induced apoptosis more in LD, diminishing the density-dependence of H2O2-induced apoptosis without altering that of spontaneous apoptosis. Akt level and activity and Hsp72 level were increased in HD, but decreased in MD and LD cells, by H2O2, and the active p38 MAPK was enhanced by H2O2 in all groups. CONCLUSION: Spontaneous apoptosis is cell density-dependent whereas H2O2-induced apoptosis is reverse density-dependent. PI3K/Akt, p38 MAPK and Hsp72 pathways are critical for the opposite density-dependence of apoptosis.
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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 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
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".