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Currently available strategies to evaluate the effectiveness of CAR T cells include conventional in vitro methods, such as cytokine secretion, cytotoxicity, proliferation, ratio of CD4/CD8 cells, and long-term killing assays, as well as in vivo mouse models.
The culture of human umbilical vein endothelial cells (HUVECs) in vitro proved that the cell proliferation ratio of both the aminolyzed and the biomacromolecules-immobilized PU membranes was improved greatly comparing with the control PU.
In contrast to what could have been expected, proliferation ratio at E9.5 was 2.8 times higher in the median anlage of Hes1−/− mutants than in wild type thyroids.
Further, proliferation ratio did not differ in mutant and wild type mice from E11.5 onwards, in accordance with constant size reduction ratio between thyroids of mutants vs. wild type until E16.5.
Consistent with the observed decrease in tumor growth, the proliferation ratio of LNCaP and C4-2 tumors was also decreased, as quantified by PCNA staining in tumors electroporated with miR-143 (Fig. 4D).
The results are expressed as CFDA-SE proliferation ratio, calculated by the proliferative response observed in co-cultures of Pv-AMA-1-stimulated PBMCs with CD4+CD25+ T cells over Pv-AMA-1-stimulated PBMCs only.
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No significant differences in the proliferation ratios were observed between the CIS cells adjacent to seminomas compared with CIS cells adjacent to non-seminomas (data not shown).
This allowed us to compare the background-corrected cpm values as well as the proliferation ratios under the two different conditions of in vitro stimulation.
We observed the inverse relationship between steroid receptor expression and proliferation in our in vitro model (proliferation ratios increased and simultaneously the number of ER, PR positive cells decreased).
Since total isotope incorporation by PBMCs and PBMC-SF cell co-cultures as well as background cpm values (conditions "c" and "d") varied from patient to patient, we calculated the background-corrected proliferation ratios for the 9 patients tested.
However, patients with DOA+ phenotypes had significantly higher proliferation ratios in COX-negative muscle fibres when compared with those with isolated optic nerve involvement, indicating a possible important disease mechanism.
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