Potential for improvement of population diet through reformulation of commonly eaten foods

Food reformulation: Reformulation of foods is considered one of the key options to achieve population nutrient goals. The compositions of many foods are modified to assist the consumer bring his or her daily diet more in line with dietary recommendations. Initiatives on food reformulation: Over the past few years the number of reformulated foods introduced on the European market has increased enormously and it is expected that this trend will continue for the coming years. Limits to food reformulation: Limitations to food reformulation in terms of choice of foods appropriate for reformulation and level of feasible reformulation relate mainly to consumer acceptance, safety aspects, technological challenges and food legislation. Impact on key nutrient intake and health: The potential impact of reformulated foods on key nutrient intake and health is obvious. Evaluation of the actual impact requires not only regular food consumption surveys, but also regular updates of the food composition table including the compositions of newly launched reformulated foods

<i>A. fumigatus</i> hyphae upregulate pro-IL-1β transcription and induce IL-1β secretion in monocytes.

<p>One million THP-1 cells/ml were treated with 10 ng/ml of LPS with and without nigericin, spores or HFs at an MOI = 10 for 6 hours (A, B), or spores for 12 hours and HFs for 6 hours (C, D). (<b>A</b>) Intracellular IL-1β gene transcription was quantified by real-time PCR and compared to control. (<b>B, C</b>) The amount of secreted IL-1β was quantified by ELISA. (<b>D</b>) TNFα secretion was measured in supernatants by ELISA. Error bars represent standard deviation of at least three separate experiments. * <i>p</i><0.05; ** <i>p</i><0.01; *** <i>p</i><0.001, compared to infected untreated cells.</p

Syk kinase signaling provides the stimulus for both IL-1β synthesis and caspase-1 activation during <i>A. fumigatus</i> infection.

<p>THP-1 cells were pretreated with 1 µM of the Syk kinase inhibitor (Syk I) for 30 min prior to challenge with HFs, and (<b>A</b>) mature IL-1β and (<b>B</b>) active caspase-1 p20 subunit were measured by ELISA. MyD88 and Syk were stably silenced by RNA interference using shRNA. (<b>C</b>) Transcript levels of pro-IL-1β in MyD88 KD and Syk KD cells treated with HFs was measured using real-time PCR. Representative real-time PCR values representative of three independent experiments are shown. The secretion (<b>D</b>) of IL-1β and (<b>E</b>) caspase-1 p20 into the supernatants of MyD88 KD and Syk KD cells treated with HFs was assessed by ELISA. All values are representatives of at least three independent experiments. Error bars represent standard deviation of at least three separate experiments. * <i>p</i><0.05; ** <i>p</i><0.01; *** <i>p</i><0.001, compared to infected untreated cells.</p

The inflammatory response against <i>A. fumigatus</i> is impaired in immunosuppressed monocytes.

<p>(<b>A</b>) THP-1 cells were stimulated for 10 min with 30 µM β-methasone prior to stimulation with HFs for 6 hours. IL-1β secretion was measured by ELISA. (<b>B</b>) THP-1 cells were stimulated for 10 min with 30 µM β-methasone prior to stimulation with 10 ng/ml LPS for 6 hours. IL-1β mRNA was quantified by real-time PCR.</p

<i>A. fumigatus</i> induced-caspase-1 activation depends on ROS production and K⁺ efflux.

<p>THP-1 cells were incubated with HFs for 6 hours in the presence or absence of 130 mM KCl, 25 mM NAC, 100 µM caspase-1/caspase-5 inhibitor (Z-WEHD-FMK), or pretreated for 30 min with 1 µM of Syk kinase inhibitor (Syk I). (<b>A inset</b>) Caspase-1 activation was analyzed by Western blot, using an antibody against the Caspase-1 p20 cleavage product. Each band intensity was measured by NIH ImageJ software (Ctrl = 1, HF = 4.848, HF + Z-WEHD-FMK = 2.92, HF + Syk I = 1.67, and HF + NAC = 1.54). (<b>A</b>) Secreted Caspase-1 p20 and (<b>B, C</b>) mature IL-1β p17 in the supernatant of infected cells, compared to the control, was assessed by ELISA. Error bars represent the standard deviation of at least three separate experiments. * <i>p</i><0.05; ** <i>p</i><0.01; *** <i>p</i><0.001, compared to infected untreated cells.</p

Purification of GFP-<i>Pf</i>Rab1A bodies.

<p>Samples taken from different stages of purification, starting with whole cells (top), and including a crude cell extract, a high speed (13,000 g) centrifugation pellet, and a fraction from a Percoll density gradient spanning 1.037 to 1.054 g/mL subsequently pelleted by centrifugation at 13,000 g. Scale bars are 3 μm. (B) Western blot using an antibody against GFP on the purified Percoll fraction shows a signal at a size (~55 kDa) consistent with GFP-<i>Pf</i>Rab1A even though the protein levels of the fusion are insufficient to visualize by Ponceau staining.</p

HK2 Recruitment to Phospho-BAD Prevents Its Degradation, Promoting Warburg Glycolysis by Theileria-Transformed Leukocytes

Theileria annulata infects bovine leukocytes, transforming them into invasive, cancer-like cells that cause the widespread disease called tropical theileriosis. We report that in <i>Theileria</i>-transformed leukocytes hexokinase-2 (HK2) binds to B cell lymphoma-2-associated death promoter (BAD) only when serine (S) 155 in BAD is phosphorylated. We show that HK2 recruitment to BAD is abolished by a cell-penetrating peptide that acts as a nonphosphorylatable BAD substrate that inhibits endogenous S155 phosphorylation, leading to complex dissociation and ubiquitination and degradation of HK2 by the proteasome. As HK2 is a critical enzyme involved in Warburg glycolysis, its loss forces <i>Theileria</i>-transformed macrophages to switch back to HK1-dependent oxidative glycolysis that down-regulates macrophage proliferation only when they are growing on glucose. When growing on galactose, degradation of HK2 has no effect on <i>Theileria</i>-infected leukocyte proliferation, because metabolism of this sugar is independent of hexokinases. Thus, targeted disruption of the phosphorylation-dependent HK2/BAD complex may represent a novel approach to control <i>Theileria</i>-transformed leukocyte proliferation

Proteomic analysis of purified PfRab1A bodies.

<p>Proteomic analysis of purified PfRab1A bodies.</p

GFP-<i>Pf</i>Rab1A fluorescence is found as discrete loci in living cells.

<p>A. GFP fluorescence is observed as discrete loci termed <i>Pf</i>Rab1A bodies in trophozoite and schizont stage parasites expressing GFP-<i>Pf</i>Rab1A. B. GFP-<i>Pf</i>Rab18 fluorescence is diffuse and closely associated with the nuclei. C. C-terminal HA-tagged <i>Pf</i>Rab1A is distributed equally throughout the parasite cytoplasm. D. Infection by parasites expressing GFP-<i>Pf</i>Rab1A is less efficient than parasites expressing GFP-<i>Pf</i>Rab18 from the same promoter. Asterisks show significant differences for each time (p ≤ 0.01) using Student’s unpaired two-tailed t-test. E. While not associated with the nucleus, the number of GFP-<i>Pf</i>Rab1A loci is proportional to the number of nuclei in the cell.</p

GFP-<i>Pf</i>Rab1A fluorescence in late schizonts is associated with rhoptry markers.

<p>Rhoptry markers Rap1 and Ron4 are found in trophozoites and schizonts as discrete foci that colocalize with GFP-<i>Pf</i>Rab1A fluorescence in late schizonts, but not in earlier phases of parasite development.</p