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IκBα reduced E. coli-induced histologic evidence of lung injury, with the intermediate (1 × 10) vector dose of IκBα again most effective.
In a canine study, a vector dose of 2·4 × 10 plaque forming units (pfu) infused into three haemophilia B dogs resulted in transient expression of Factor IX at supraphysiological levels.
At the lowest vector dose of HD-Ad-CMV-LacZ, 1 × 107 vp/eye, transgene expression was detected, but consistency of expression throughout the entire retina was not observed as only several areas around the retina showed β-galactosidase activity.
As an example, a vector dose of 4×10 VG/kg yielded plasma factor IX protein levels of 9 10% resulting in 55 65% activity in the human factor IX activity assay (Fig. 2A and B).
Muscle-directed AAV vectors were subsequently shown to achieve low but possibly clinically relevant plasma levels (70 ng/ml) of canine Factor IX in haemophilia B dogs at a vector dose of 8·5 × 10 vp/kg.
Haemophilia B canines treated with a vector dose of 1 2 × 10 vg/kg responded with expression of circulating canine Factor IX between 220 ng/ml and 590 ng/ml (4 12%) (Mount et al, 2002).
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C57BL/6 mice were initially injected subretinally with AAV OZ and dual AAV TS-L and hybrid AK-L vectors (dose of each vector/eye: 1.7 × 10 GC), all encoding EGFP under the transcriptional control of the CMV promoter.
One month-old wild-type C57BL/6 mice were injected with the dual AAV vectors (dose of each vector/eye: 1.7 × 10 GC) and eyecup lysates were evaluated 1 month later using Western blot analysis with anti-HA antibodies.
To determine which cell type in the retina expressed ABCA4, we used dual AAV OV vectors (dose of each vector/eye: 1 × 10 GC) that contained either the PR-specific RHO and RHOK, or the RPE-specific VMD2 promoters.
The number of lipofuscin granules in Abca4−/− RPE was normalized 3 months post subretinal injection of improved dual AAV hybrid ABCA4 vectors (dose of each vector/eye: 1 × 10 GC, Fig. 5D).
We then injected 5.5 month-old pigmented Abca4−/− mice subretinally in the temporal region of the eye with the improved dual AAV vectors (dose of each vector/eye: 1.8 × 10 GC).
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