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As a rule of thumb, the culture volume should be less or equal to 10% of the shaking flask capacity, although in our hands, protein production with culture volumes occupying 20% of the flask capacity was possible (Rosano et al., 2011)
Yet, in other cardiovascular diseases, the connection between inflammation and increased vascular stiffness has been better characterized: here, inflammatory processes have been shown to promote arterial stiffening through a variety of mechanisms, including the induction of endothelial dysfunction and BM stiffening, increased proliferation of SMCs (49)resulting in arterial wall thickening and reduced complianceand remodeling and stiffening of the ECM in different segments of the arterial wall
2001,000 mcg once daily, titrated upward gradually over an ~10-week course in roughly 200 mcg steps

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