Genetic characterization of glucose transporter function in Leishmania mexicana

Both insect and mammalian life cycle stages of Leishmania mexicana take up glucose and express all three isoforms encoded by the LmGT glucose transporter gene family. To evaluate glucose transporter function in intact parasites, a null mutant line has been created by targeted disruption of the LmGT locus that encompasses the LmGT1, LmGT2, and LmGT3 genes. This Δlmgt null mutant exhibited no detectable glucose transport activity. The growth rate of the Δlmgt knockout in the promastigote stage was reduced to a rate comparable with that of WT cells grown in the absence of glucose. Δlmgt cells also exhibited dramatically reduced infectivity to macrophages, demonstrating that expression of LmGT isoforms is essential for viability of amastigotes. Furthermore, WT L. mexicana were not able to grow as axenic culture form amastigotes if glucose was withdrawn from the medium, implying that glucose is an essential nutrient in this life cycle stage. Expression of either LmGT2 or LmGT3, but not of LmGT1, in Δlmgt null mutants significantly restored growth as promastigotes, but only LmGT3 expression substantially rescued amastigote growth in macrophages. Subcellular localization of the three isoforms was investigated in Δlmgt cells expressing individual LmGT isoforms. Using anti-LmGT antiserum and GFP-tagged LmGT fusion proteins, LmGT2 and LmGT3 were localized to the cell body, whereas LmGT1 was localized specifically to the flagellum. These results establish that each glucose transporter isoform has distinct biological functions in the parasite

Potential early-life predictors of dietary behaviour in adulthood: a retrospective study

This paper is in closed access.OBJECTIVE: Unnecessary dietary restraint (ie in the absence of a need to lose weight) and chronic overeating are both very unhealthy activities. As a precursor to a more involved longitudinal study, we sought to identify potential early-life predictors that merit scrutiny in this context. DESIGN: Four retrospective questionnaire studies were conducted (Study 1, N¼242; Study 2, N¼297; Study 3, N¼175; Study 4, N¼261). Female participants (18–30 y) completed measures of current dietary restraint and overeating. They also recalled experiences between 5 and 10 years of age. All were staff or students at Loughborough University (UK). RESULTS: After considering obvious sources of systematic bias, we report evidence that (i) dietary restraint is related to memories of maternal weight and dietary behaviour, and (ii) overeating and meal-size selection are both associated with memories of receiving a high-energy diet. CONCLUSION: The role of maternal factors in dietary restraint is consistent with previous research exploring the early onset of this behaviour. However, the relationship between childhood diet and overeating has not been suggested elsewhere. This is particularly important because it suggests a previously unreported correspondence between childhood experience and behaviours associated with obesity in adulthood