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In a recent paper by Pascaud et al [11], a three-layered gradient of 1.10, 1.15 and 1.30 g/ml (2 ml of each) was produced by dilution of OptiPrep with 0.25 M sucrose, 6 mM EDTA, 60 mM Tris-HCl, pH 7.4 and the soil suspension (5 ml) layered on top

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The evidence reviewed herein does not yet support formal treatment guidelines, given the predominance of low-level data and the absence of head-to-head trials in MC4R -deficient populations

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