Project description:Overfeeding reduces laying performance in broiler breeder hens (Gallus gallus domesticus). To unravel the effect of feeding regimes on energy metabolism and egg production, high-throughput RNA sequencing was utilized to identify differentially expressed genes (DEGs) in ovary, liver and adipose tissues of broiler chickens under ad libitum and restricted feeding. The transcriptome analysis showed that 289, 388 and 204 DEGs were identified in the adipose tissue, liver and ovary, respectively, between ad libitum and restricted feeding groups. Bioinformatics analysis by STRING further revealed DEGs were significantly enriched in phagosome pathway, lipid transport and location biological process, and the molecular function of lipid transporter activity and nutrient reservoir activity in ovary; the pathways of “steroid hormone biosynthesis” and “metabolism of xenobiotics by cytochrome P450”, and the molecular function of nutrient reservoir activity in adipose tissue; the metabolic pathways, Jak-STAT signaling pathway and PPAR signaling pathway in liver.
Project description:Brassica rapa is grown in northwestern Spain to obtain turnip greens. The tops of the same plants (flower stems with buds) are cut and sell as turnip tops, increasing the value of the crop. This practice could be extended to other brassicas. The objectives of this work are to study the phytochemical potential of tops of coles (Brassica oleracea) and leaf rape (Brassica napus) compared to turnip tops and to compare tops of different coles (cabbage, kale, tronchuda cabbage), which differ in their morphology and use. We evaluated the content of glucosinolates and phenolic compounds and the antioxidant capacity in leaves and tops of the three species. We found that tops had higher amount of glucosinolates than leaves. Phenolic content and antioxidant capacity followed the opposite trend. Therefore, consumption of leaves and tops are complementary, since both type of organs are enriched with different types of compound. Local varieties of kale, curly kale, cabbage and curly leave cabbage are interesting because of their GSLs and phenolic content and antioxidant capacity in both leaves and tops. From the human health perspective, tops of coles and leaf rape are interesting as new crops to include in the diet.
Project description:Small peptides with amino acid sequences similar to native skin proteins can beneficially affect clinical appearance (wrinkles) and architecture (collagen and elastic fibre deposition and epidermal thickness). However, the discovery of new cosmetic peptides has not been underpinned by any guiding hypothesis. As endogenous extracellular matrix (ECM)-derived peptides (matrikines) produced during tissue remodelling can influence cell activity, we hypothesised that protease cleavage site prediction can identify putative novel matrikines. This RNA-seq data is the in vivo test part of an in silico to in vivo discovery pipeline, which enables the prediction and characterisation of peptide matrikines with therapeutic potential. We use this pipeline to identify two novel ECM peptides (GPKG and LSVD) which, in combination, act in vitro to enhance the transcription of ECM organisation and cell proliferation genes and in vivo to promote epithelial and dermal remodelling. This pipeline approach can both identify new matrikines and provide insights into the mechanisms underpinning tissue repair.
Project description:Small peptides with amino acid sequences similar to native skin proteins can beneficially affect clinical appearance (wrinkles) and architecture (collagen and elastic fibre deposition and epidermal thickness). However, the discovery of new cosmetic peptides has not been underpinned by any guiding hypothesis. As endogenous extracellular matrix (ECM)-derived peptides (matrikines) produced during tissue remodelling can influence cell activity, we hypothesised that protease cleavage site prediction can identify putative novel matrikines. This RNA-seq data is the in vitro testing part of an in silico to in vivo discovery pipeline, which enables the prediction and characterisation of peptide matrikines with therapeutic potential. We use this pipeline to identify two novel ECM peptides (GPKG and LSVD) which, in combination, act in vitro to enhance the transcription of ECM organisation and cell proliferation genes and in vivo to promote epithelial and dermal remodelling. This pipeline approach can both identify new matrikines and provide insights into the mechanisms underpinning tissue repair.
Project description:Small peptides with amino acid sequences similar to native skin proteins can beneficially affect clinical appearance (wrinkles) and architecture (collagen and elastic fibre deposition and epidermal thickness). However, the discovery of new cosmetic peptides has not been underpinned by any guiding hypothesis. As endogenous extracellular matrix (ECM)-derived peptides (matrikines) produced during tissue remodelling can influence cell activity, we hypothesised that protease cleavage site prediction can identify putative novel matrikines. This RNA-seq data is the in vitro testing part of an in silico to in vivo discovery pipeline, which enables the prediction and characterisation of peptide matrikines with therapeutic potential. We use this pipeline to identify two novel ECM peptides (GPKG and LSVD) which, in combination, act in vitro to enhance the transcription of ECM organisation and cell proliferation genes and in vivo to promote epithelial and dermal remodelling. This pipeline approach can both identify new matrikines and provide insights into the mechanisms underpinning tissue repair.
Project description:Quorum sensing (QS) in bacteria has been a well studied cellular communication phenomenon for decades. In recent years, such systems have been repurposed for the use of biosensors in both cellular and cell-free contexts as well as for inducible protein expression in non-traditional chassis organisms. Such biosensors are particularly intriguing when considering the association between the pathogenesis of some bacteria and the QS signaling intermediates. Considering this relationship, and considering the recent demonstration of the species L. plantarum WCFS1 as both a synthetic biology chassis and an organism capable of detecting a pathogen-associated QS molecule, we wanted to develop this organism as a QS sentinel. Using an approach combining techniques from both systems and synthetic biology, we identified a number of native QS response genes and altered associated promoter activity to tune the output of L. plantarum cultures exposed to N-3-oxododecanoyl homoserine lactone. The resulting engineered QS sentinel reinforces the potential of modified Lactic Acid Bacteria for use in human health promoting applications, and also demonstrates a simple rational workflow to engineer sentinel organisms to respond to any environmental or chemical stimuli.
Project description:3’,5’-cyclic adenosine monophosphate (cAMP) is an ubiquitous second messenger which regulates multiple physiological functions by acting in distinct subcellular microdomains. Although several targeted cAMP biosensors have been developed and used in cell lines or neonatal cardiomyocytes, it is unclear whether such biosensors can be successfully used in vivo, especially in the context of disease. Here, we generated the first transgenic mouse model expressing a targeted cAMP sensor (Epac1-PLN) and analyzed microdomain-specific second messenger dynamics in the vicinity of the sarcoplasmic reticulum calcium ATPase (SERCA). We demonstrate the biocompatibility of this targeted sensor and its potential for real-time monitoring of compartmentalized cAMP signaling in adult cardiomyocytes isolated from healthy mouse heart and from in vivo cardiac disease model. Finally, we used gene arrays to verify that the mRNA expression levels of the main players involved in cAMP signaling such as beta-ARs, G-protein, adenylyl cyclases, PKAs, PDEs, and Epac were not significantly changed between wildtype and transgenic Epac1-PLN expressing cardiomyocytes. To compare cardiac specific mRNA expression between wildtype and Epac1-PLN transgenic mice, cardiomyocyte RNA from total of 5 individual wildtype and 5 Epac1-PLN transgenic mice was isolated and submitted for the Illumina gene array analysis to the Göttingen transcriptome analysis laboratory (TAL).
Project description:In this study, the researchers isolated a colorless waxy solid from the chloroform extract of Allium stipitatum as Compound 1. They demonstrated compound 1 is a potent bactericidal agent against nonreplicating Mycobacterium tuberculosis. Using microarray technology the authors report gene expression profiles in cells treated with either 2, 5, or 10 ug/ml of compound 1 or an equivalent amount of DMSO as control for 6 h. Gene expression studies revealed that transcriptional profiles elicited in response to compound 1 were similar to the profiles generated during treatment of cells with compounds such as menadione and 8-quinolinol that result in oxidative stress. They included the thioredoxin system components encoded by trxB2 and trxC as well as several genes associated with the heat shock response such as clpB, sigH, dnaJ, dnaK, hsp, Rv0331, Rv3463, Rv3054c, and Rv1334-Rv1335. These results suggest that compound 1 possibly generates damaged proteins and other oxidative stress signals as part of its mechanism of action. The following is the full abstract of this published study. O'Donnell G, et al. (2009) J Nat Prod 72(3):360-365 From Allium stipitatum, three pyridine-N-oxide alkaloids (1-3) possessing disulfide functional groups were isolated. The structures of these natural products were elucidated by spectroscopic means as 2-(methyldithio)pyridine-N-oxide (1), 2-[(methylthiomethyl)dithio]pyridine-N-oxide (2), and 2,2'-dithio-bis-pyridine-N-oxide (3). The proposed structure of 1 was confirmed by synthetic S-methylthiolation of commercial 2-thiopyridine-N-oxide. Compounds 1 and 2 are new natural products, and 3 is reported for the first time from an Allium species. All compounds were evaluated for activity against fast-growing species of Mycobacterium, methicillin-resistant Staphylococcus aureus, and a multidrug-resistant (MDR) variants of S. aureus. Compounds 1 and 2 exhibited minimum inhibitory concentrations (MICs) of 0.5-8 mug/ml against these strains. A small series of analogues of 1 were synthesized in an attempt to optimize antibacterial activity, although the natural product had the most potent in vitro activity. In a whole-cell assay at 30 mug/ml, 1 was shown to give complete inhibition of the incorporation of (14)C-labeled acetate into soluble fatty acids, indicating that it is potentially an inhibitor of fatty acid biosynthesis. In a human cancer cell line antiproliferative assay, 1 and 2 displayed IC(50) values ranging from 0.3 to 1.8 muM with a selectivity index of 2.3 when compared to a human somatic cell line. Compound 1 was evaluated in a microarray analysis that indicated a similar mode of action to menadione and 8-quinolinol by interfering with the thioredoxin system and up-regulating the production of various heat shock proteins. This compound was also assessed in a mouse model for in vivo toxicity. A dose response design type examines the relationship between the size of the administered dose and the extent of the response of the organism(s). Compound Based Treatment: Allium stipitatum (garlic) extract dose_response_design
Project description:The median eminence (ME) of the hypothalamus is a structure that rapidly adapts to nutrient availability. We have used single-cell RNA sequencing to characterize the transcriptional profiles of cells specifically in the median eminence in the fasted and refed states. In our study, we focus on characterizing the oligodendrocyte population of the ME, and identifying how subpopulations respond to nutrient availability.