In Vitro Glycosylation of the Membrane Protein γ-Sarcoglycan in Nanodiscs.

Membrane glycoproteins are proteins that reside in the membranes of cells and are post-translationally modified to have sugars attached to their amino acid side chains. Studies of this subset of proteins in their native states are becoming more important since they have been linked to numerous human diseases. However, these proteins are difficult to study due to their hydrophobic nature and their propensity to aggregate. Using membrane mimetics allows us to solubilize these proteins, which, in turn, allows us to perform glycosylation in vitro to study the effects of the modification on protein structure, dynamics, and interactions. Here, the membrane glycoprotein γ-sarcoglycan was incorporated into nanodiscs composed of long-chain lipids and membrane scaffold proteins to perform N-linked glycosylation in which an enzyme attaches a sugar to the asparagine side chain within the glycosylation site. We previously performed glycosylation of membrane proteins in vitro when the protein had been solubilized using different detergents and short-chain lipids. This work demonstrates successful glycosylation of a full-length membrane protein in nanodiscs providing a more biologically relevant sample to study the effects of the modification.

Statewide Payment and Delivery Reform: Do States Have What It Takes?

States' role in payment as well as coverage will be subject to debate as the administration and the Congress decide how to address the Affordable Care Act (ACA) and otherwise reshape the nation's health policies. Acting as stewards of health care for the entire state population and stimulated by concern about rising costs and federal support under the ACA, the elected and administrative leaders of some states have been using their political influence and authority to improve their state's overall systems of care regardless of who pays the bill. In early 2015 we conducted on-site interviews with key stakeholders in five states to explore their strategies for payment and delivery reform. We found that despite these states' similar goals, differences in their statutory authority and purchasing power, along with their leaders' willingness to use them, significantly influence a state's ability to achieve reform objectives. We caution federal and state policy makers to recognize the reality that state leaders' political desire to exercise stewardship may not be enough to achieve it.

Historical peat loss explains limited short-term response of drained blanket bogs to rewetting.

This study assessed the short-term impacts of ditch blocking on water table depth and vegetation community structure in a historically drained blanket bog. A chronosequence approach was used to compare vegetation near ditches blocked 5 years, 4 years and 1 year prior to the study with vegetation near unblocked ditches. Plots adjacent to and 3 m away from 70 ditches within an area of blanket bog were assessed for floristic composition, aeration depth using steel bars, and topography using LiDAR data. No changes in aeration depth or vegetation parameters were detected as a function of ditch-blocking, time since blocking, or distance from the ditch, with the exception of non-Sphagnum bryophytes which had lower cover in quadrats adjacent to ditches that had been blocked for 5 years. Analysis of LiDAR data and the observed proximity of the water table to the peat surface led us to conclude that the subdued ecosystem responses to ditch-blocking were the result of historical peat subsidence within a 4-5 m zone either side of each ditch, which had effectively lowered the peat surface to the new, ditch-influenced water table. We estimate that this process led to the loss of around 500,000 m3 peat within the 38 km2 study area following drainage, due to a combination of oxidation and compaction. Assuming that 50% of the volume loss was due to oxidation, this amounts to a carbon loss of 11,000 Mg C over this area, i.e. 3 Mg C ha-1. The apparent 'self-rewetting' of blanket bogs in the decades following drainage has implications for their restoration as it suggests that there may not be large quantities of dry peat left to rewet, and that there is a risk of inundation (potentially leading to high methane emissions) along subsided ditch lines. Many peatland processes are likely to be maintained in drained blanket bog, including support of typical peatland vegetation, but infilling of lost peat and recovery of original C stocks are likely to take longer than is generally anticipated.