A lentiviral vector bearing a reverse intron demonstrates superior expression of both proteins and microRNAs.

While lentiviral expression vectors are widely used in many facets of molecular biology, due to their ability to stably express heterologous genes in both dividing and non-dividing cells, they suffer from the disadvantage that introns inserted into the vector genome are generally rapidly lost by splicing in packaging cell lines. The presence of an intron, if achievable, has the potential to facilitate the expression of transgene cDNAs, as splicing has been extensively shown to facilitate mRNA biogenesis and function. Moreover, if a stable intron could be introduced into a lentiviral vector, this could greatly facilitate the expression of microRNAs (miRNAs), and especially miRNA clusters, as the introduction of pri-miRNA stems into the exonic region of a lentiviral vector can strongly reduce both vector titer and the expression of any miRNA-linked indicator gene due to cleavage of the vector RNA genome by cellular Drosha. Here, we describe a novel lentiviral vector design in which transgenes and/or miRNAs are expressed using an antisense-orientated, inducible promoter driving an expression cassette bearing a functional intron. We demonstrate that this lentiviral vector, called pTREX, is able to express higher levels of both transgenes and pri-miRNA clusters when compared with a closely similar conventional lentiviral vector.

An exploration of how meal preparation activities relate to self-rated time pressure, stress, and health in Canada: A time use approach.

While previous work has provided a foundation for understanding the importance of the links between time use and diet, there has been little done to link time use to health outcomes. In this study, time use and self-rated health variables from the 2015 Time Use Cycle of Statistics Canada's General Social Survey are used to explore whether there are direct associations between time spent on meal preparation and health for Canadian Adults. In addition, this paper uses respondents' sequences of activities data from a time use diary to provide novel findings about the context of activities that precede and follow meal preparation. Proportional odds and logistic regression models are computed and show that there are significant relationships between spending more time on meal preparation and improved mental health and lower levels of stress. More time on meal preparation is also linked to general feelings of having less time. The analysis of activities preceding and following meal preparation activities demonstrates that individuals with different levels of self-rated stress or feelings of having extra time have significantly different activity sequence distributions (e.g., those reporting higher levels of stress are more likely to participate in chores and care activities). Exploring activity sequences related to meal preparation provides a first step in furthering the research community's grasp of the causal relationship between food-related time use and health and well-being outcome variables. Ultimately, this paper builds on the past literature on time use and meal preparation by establishing direct links between time spent on meal preparation activities, self-rated health and time use variables, in addition to offering insights into what activities surround this important activity via a novel sequence analysis.

A mole rat's gut microbiota suggests selective influence of diet on microbial niche space and evolution.

IMPACT STATEMENT: The composition of the microbiota is of critical importance for health and disease, and is receiving increased scientific and medical scrutiny. Of particular interest is the role of changing diets as a function of agriculture and, perhaps to an even greater extent, modern food processing. To probe the connection between diet and the gut's microbial community, the microbiota from a mole rat, a rodent with a relatively unusual diet, was analyzed in detail, and the microbes found were compared with previously identified organisms. The results show evidence of an adaptive radiation of some microbial clades, but relative stability in others. This suggests that the microbiota, like the genome, carries with it housekeeping components as well as other components which can evolve rapidly when the environment changes. This study provides a very broad view of the niche space in the gut and how factors such as diet might influence that niche space.