Developmental Regulation of Dirofilaria Immitis Microfilariae and Evaluation of Ecdysone Signaling Pathway Transcript Level Using Droplet Digital Polymerase Chain Reaction
Bibliographic record
Abstract
Abstract BackgroundCurrent Dirofilaria immitis prevention relies on macrocyclic lactones but evidence of drug-resistant isolates has called for the need for alternative approaches to disease intervention. As microfilariae are known to be in a state of developmental arrest in their mammalian host and then undergo a rapid molt once inside the arthropod, this study aimed to look at the developmental regulation of D. immitis microfilariae that occurs in their arthropod host using in vitro approaches and to investigate the role of the ecdysone signaling system in this development.MethodsD. immitis microfilariae extracted from dog blood were incubated in various media conditions to identify suitable conditions for in vitro culture and development, and to determine the effects of FBS, mosquito cells, and ecdysteroid on the development of the microfilariae. Transcript level of the ecdysone signaling pathway components were measured with droplet digital PCR.ResultsIn vitro conditions that best promote early development of D. immitis microfilariae to the “late sausage stage” have been identified. FBS showed inhibitory effects on the development and motility of the microfilariae, but media conditioned with Anopheles gambiae cells was favorable to microfilarial growth. Transcript level study using droplet digital PCR also showed that developing microfilariae displayed upregulation of the ecdysone signaling system components, and 20‑hydroxyecdysone increased the proportion of larvae developing to the sausage and late sausage stages in vitro.ConclusionsThe arthropod host environment provides cues required for the rapid development of the D. immitis microfilariae and the ecdysone signaling system may play an important role in filarial nematode developmental transitions. This study contributes to a better understanding the developmental process of D. immitis microfilariae.
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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 | 0.000 |
| 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".