Crystal structures and kinetic studies of a laboratory evolved aldehyde reductase explain the dramatic shift of its new substrate specificity.

The Fe2+-dependent E. coli enzyme FucO catalyzes the reversible interconversion of short-chain (S)-lactaldehyde and (S)-1,2-propanediol, using NADH and NAD+ as cofactors, respectively. Laboratory-directed evolution experiments have been carried out previously using phenylacetaldehyde as the substrate for screening catalytic activity with bulky substrates, which are very poorly reduced by wild-type FucO. These experiments identified the N151G/L259V double mutant (dubbed DA1472) as the most active variant with this substrate via a two-step evolutionary pathway, in which each step consisted of one point mutation. Here the crystal structures of DA1472 and its parent D93 (L259V) are reported, showing that these amino acid substitutions provide more space in the active site, though they do not cause changes in the main-chain conformation. The catalytic activity of DA1472 with the physiological substrate (S)-lactaldehyde and a series of substituted phenylacetaldehyde derivatives were systematically quantified and compared with that of wild-type as well as with the corresponding point-mutation variants (N151G and L259V). There is a 9000-fold increase in activity, when expressed as kcat/KM values, for DA1472 compared with wild-type FucO for the phenylacetaldehyde substrate. The crystal structure of DA1472 complexed with a non-reactive analog of this substrate (3,4-dimethoxyphenylacetamide) suggests the mode of binding of the bulky group of the new substrate. These combined structure-function studies therefore explain the dramatic increase in catalytic activity of the DA1472 variant for bulky aldehyde substrates. The structure comparisons also suggest why the active site in which Fe2+ is replaced by Zn2+ is not able to support catalysis.

Offspring migration and nutritional status of left-behind older adults in rural China.

Improvements in nutritional status is a principal pathway to good health. This study examines the effect of migration of adult children on the nutrient intake of left-behind older adults in rural China. We use data from four waves (2004-2011) of the China Health and Nutrition Survey and utilize individual fixed effects methods to panel data. Results show that the migration of offspring is associated with significantly higher nutritional status of their left-behind parents, especially higher intake of proteins, carbohydrates, vitamins B1-B3, phosphorus, magnesium, iron, selenium, and copper. The intake of some of these nutrients is below recommended levels. The magnitude of the estimated effects vary between 4% and 24 %. Older adults who live with their grandchildren in rural households or have a low income benefit more from having adult child migrants in the household. The improvement of nutrition outcomes of left-behind older adults is mainly due to increased consumption of cereals, meat, eggs, and fish.

Life cycle development of obesity and its determinants in six European countries.

This paper empirically examines the effect of parents' and individuals' own socioeconomic status on overweight and obesity, and investigates how this effect changes over the life cycle. The impact of individuals' health behaviours on their obesity status later in life is also studied. We use data from Denmark, Finland, France, Greece, the Netherlands and the U.K. in which 4595 individuals aged 50-65 are surveyed and where individuals' height and weight at different ages (25, 35, 45 and current age) are available. We perform "repeated cross-sections" analyses as well as dynamic probit analyses of the individuals' obesity histories. We contribute to the literature by examining the role of a variety of obesity determinants over the whole life cycle, not only over a certain portion of individuals' lives. Key findings are: (i) parents' socioeconomic status predicts obesity in early adulthood whereas the individual's own socioeconomic status as adult is more important in explaining obesity at later stages of the life cycle, (ii) changes in obesity status are associated with changes in health behaviours, (iii) obesity in late adulthood is strongly and positively correlated with overweight and obesity in younger ages, and (iv) cross-country differences in obesity and overweight largely remain after controlling for parental and childhood factors and individuals' health behaviours.