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RT-qPCR was also used to assess HES1 mRNA levels in OSA tumor and matched normal bone samples.
Average HES1 mRNA expression was elevated 2.57-fold in canine OSA tumors compared to the matched normal bone; however, this fold change was highly variable when each OSA tumor was compared to its matched normal bone sample, with 5 tumors exhibiting elevated expression compared to normal bone and 4 tumors having virtually unchanged expression.
For example, HES1 mRNA expression in tumors relative to normal bone was elevated in 5 of 9 canine tumors relative to matched normal bone samples in our study and 6 of 10 human tumors in the Tanaka study [ 25].
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Nine additional appendicular OSA tumor samples were collected from which matched normal metaphyseal bone was harvested from the same limb (at least one joint space away from the tumor) following amputation.
Approximately 52 elderly male patients with osteoporosis, 50 elderly men with osteopaenia, and 31 age-matched normal bone mass controls participated in the study.
Using a unique quartet design, we sequenced matched normal (blood following remission) and tumor (bone marrow at diagnosis) patient samples, and the parents of each case.
DNA and RNA were extracted from frozen tumor tissue while matched normal DNA was extracted from peripheral blood, remission bone marrow, histologically normal tissue, and sorted T-cells (Supplementary Data 1).
We defined differentially spliced events as those exhibiting a difference in isoform ratio of ≥10%% between a solid tumor sample and its matched normal, or alternately between an acute myeloid leukemia sample and the median across four normal bone marrow samples.
In 123 osteoporotic cases and 131 matched normal controls, Langdahl et al. found that the same 713-8delC deletion was associated with low bone mass and increased bone turnover [ 17].
Hundred osteoporotic men with age matched normal were studied for serum levels of sex steroids, PTH, IGF system components, cytokines and bone turnover markers.
All the cases had matched normal tissues.
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