Project description:Background: Poultry eggs are a low cost, high protein nutrient package that can be consumed as part of a quality diet. However, consumption of poultry egg products has been historically contentious, which highlights the importance of investigating impacts of long-term egg consumption on metabolic health. Objective: Our study utilized the zebrafish Danio rerio, a newly-defined model of human metabolic health, to understand the metabolic consequence of consuming egg products in lieu of other well-described protein sources. Methods: Reference diets were formulated to contain multi-source protein with casein and fish protein hydrolysate (CON; control protein sources), the protein sources of which have been vetted in numerous reference diets. These proteins were then replaced in part with either whole egg (WE; protein and lipid source), egg white (EW; protein source), wheat gluten (WG; cereal protein source), or a high lipid content diet containing a multi-source protein with casein and fish protein hydrolysate (HFCON; isonitrogenous and isolipidic with the WE diet) in a 34-week trial. Daily feeding was initiated at early juvenile life stage and terminated at late reproductive adult stage. Results: The amino acid composition of control vs egg product diets did not vary substantially, although methionine and lysine were apparently limiting in fish fed WG. At termination, fish fed EW as the protein source had similar weight gain and body composition to those fed the CON diet. Blood glucose, both fasting and postprandial, did not differ among any dietary treatment. Assessment of the liver transcriptome using RNAseq revealed no differential gene expression between zebrafish fed CON or WE diets. Zebrafish fed WG had lower weight gain in males. Conclusions: Long-term consumption of egg products promoted metabolic health equal to that of historically-relevant proteins. These data support the value of egg products for maintaining long-term metabolic health in animal diets.
Project description:Previous work has demonstrated that elevated maternal lipid intake (particularly from dairy products) is associated with increased lipids and altered fatty acid profile in milk produced by healthy lactating women. We investigate our primary hypothesis that a maternal diet rich in full-fat dairy products would simultaneously increase milk lipid percent and expression of genes related to the uptake and/or de novo biosynthesis of milk lipids. Within a a randomized, crossover study we performed a microarray analysis comparing 6 lactating women with diets rich in full-fat or non-fat dairy products. Each woman consumed one of the two diets during timepoint one, and after a two week washout period, switched over to the other diet. The timepoint value indicates whether the particular dietary treatment was consumed first or second.
Project description:The proteomic profiles of silky fowl egg yolk (SFEY) and Leghorn egg yolk (LEY) were analysed by bottom-up label-free liquid chromatography tandem-mass spectrometry (LC-MS/MS), aiming to provide a theoretical basis for understanding the proteomic and biological differences between the two yolks and further develop the nutritional and biomedical value of silky fowl eggs.
Project description:Eggless/SETDB1 (Egg), the only essential histone methyltransferase (HMT) in Drosophila, plays a role in gene repression, including piRNA‐mediated transposon silencing in the ovaries. Previous studies suggested that Egg is post‐translationally modified and showed that Windei (Wde) regulates Egg nuclear localization through protein–protein interaction. Monoubiquitination of mammalian SETDB1 is necessary for the HMT activity. Here, using cultured ovarian somatic cells, we show that Egg is monoubiquitinated and phosphorylated but that only monoubiquitination is required for piRNA‐mediated transposon repression. Egg monoubiquitination occurs in the nucleus. Egg has its own nuclear localization signal, and the nuclear import of Egg is Wde‐independent. Wde recruits Egg to the chromatin at target gene silencing loci, but their interaction is monoubiquitin‐independent. The abundance of nuclear Egg is governed by that of nuclear Wde. These results illuminate essential roles of nuclear monoubiquitination of Egg and the role of Wde in piRNA‐mediated transposon repression.
Project description:During animal development, a fertilized egg is initially under the control of maternal products and only starts zygotic transcription after several cell divisions. In animals such as Xenopus, zebrafish and Drosophila, a massive increase in zygotic transcription occurs during the mid-blastula transition (MBT), when cells shift from rapid, synchronous cell divisions without gap phases to prolonged asynchronous divisions. Before MBT, only a few so-called pre-MBT genes are expressed. How transcription is set up during these early stages is poorly understood. For example, paused RNA Polymerase (Pol II) is frequently found at developmental control genes in mammalian embryonic stem cells and Drosophila embryos but when Pol II pausing is first established in the embryo is unknown. We have analyzed the genome-wide Pol II occupancy during the maternal-to-zygotic transition in hand-staged Drosophila embryos. The results show that massive Pol II recruitment and pausing is established during MBT. The ~100 genes that are transcribed before MBT are particularly short, consistent with a need for rapid transcription during these early cell divisions. Remarkably, most of these genes are transcribed without Pol II pausing and this correlates with a TATA-enriched promoter type. This suggests that distinct strategies are used for activation in the early Drosophila embryo and this may reflect general dynamic properties of promoters used throughout development. Mnase-seq in staged Drosophila embryos
Project description:Transfer of somatic cell nuclei to enucleated eggs or ectopic expression of specific transcription factors are two different reprogramming strategies used to generate pluripotent cells from differentiated cells. However, they are poorly efficient and other unknown factors might be required to increase their success rate. Here, we show that Xenopus egg extracts at the metaphase stage (M phase) have a strong reprogramming activity on mouse embryonic fibroblasts (MEFs). First, they reset replication properties of MEF nuclei toward a replication profile characteristic of early development and they erase several epigenetic marks, such as trimethylation of H3K9, H3K4 and H4K20. Second, when MEFs are reversibly permeabilized in the presence of M phase Xenopus egg extracts, they show a transient increase in cell proliferation, form colonies and start to express specific pluripotency markers. Finally, transient exposure of MEF nuclei to M phase Xenopus egg extracts increases the success of nuclear transfer to enucleated mouse oocytes and strongly synergize with the production of pluripotent stem cells by ectopic expression of transcription factors. The mitotic stage of the egg extract is crucial as neither of these effects is detected when using interphasic Xenopus egg extracts. Our data demonstrate that mitosis is essential to make mammalian somatic nuclei prone to reprogramming and that, surprisingly, the heterologous Xenopus system has features that are conserved enough to remodel mammalian nuclei.
Project description:Multiple mechanisms likely contribute to the increase in chromosome missegregation that leads to production of aneuploid eggs and fetuses at advanced maternal age. It is therefore considered unlikely that a single approach could prevent age-related egg aneuploidy. Here we show using three independent approaches that ovulation reduction is sufficient to prevent egg aneuploidy in aged mammals. To gain insights into the mechanism underlying the rescue in egg aneuploidy, we show that ovulation suppression correlates with retention of chromosomal Rec8-cohesin, implying that ovulations are linked to cohesin deterioration. Moreover, we discovered that ageing alters 3D chromatin organization by single-nucleus Hi-C (snHi-C). Extruded loops increase in size with age and this is retarded by ovulation reduction. We conclude that reducing ovulations leads to retention of chromosomal Rec8, which maintains interphase chromatin structure and promotes chromosome segregation and production of euploid eggs. Importantly, our data suggest that ovulation itself contributes to the maternal age effect. This work provides the first experimental evidence that progesterone treatment reduces egg aneuploidy and suggests that hormonal contraception can reduce the risk of trisomic pregnancies like Down’s syndrome at advanced maternal age.
Project description:Even though the incidence of salmonellosis in humans has decreased over the last years, Salmonella spp. are still a leading cause of foodborne outbreaks in Europe (Anon., 2014). Of more than 2500 different serovars of Salmonella enterica, S. enterica serovar Enteritidis (S. Enteritidis) is the most frequently reported serovar in relation to food borne disease, and egg and egg products are the most important vehicles (Anon., 2014). It has recently been shown that S. Enteritidis is superior to other serovars tested regarding survival in egg white, which may explain why many egg borne outbreaks are caused by this serovar (De Vylder et al., 2013). The genetic background for this apparent better adaptation to survival in egg is only partially known. The aim of this work was to carry out gene expression analysis in order to understand how S. Enteritidis adapts to growth in the hostile environment of egg. This study analyzed gene expression of this bacterium during growth in whole egg, and whether highly expressed genes were essential for the growth. High quality RNA was extracted from S. Enteritidis using an improved modified RNA-extraction protocol. Global gene expression during growth in whole egg was compared to growth in LB-medium using DNA array method. Twenty-six genes were significantly upregulated during growth in egg; these belonged to amino acids biosynthesis, di/oligopeptide transport system, biotin synthesis, ferrous iron transport system, and type III secretion system. Significant downregulation of 15 genes related to formate hydrogenlyase (FHL) and trehalose metabolism was observed. The results suggested that S. Enteritidis is starved for amino-acids, biotin and iron when growing in egg.