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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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How does orotic acid affect the growth of R.etli CE3?

ABSTRACT: Growth inhibition of Rhizobium etli and other organisms by exogenous OA has not been previously reported. We conductedÂ genome-wide transcriptomic analysisÂ to determine the possible mechanism by which OA exerts its growth-inhibitory effect on R. etli CE3. The observed changes fall into several broad categories. First, the decreased expression of genes involved in protein synthesis (including ribosomal genes), replication and energy production indicates that the cells arrest or at least slow their growth, which agrees with the observed growth arrest induced by OA.Â The upregulation of a key enzyme (PrsAch) forÂ 5'-phosphoribosyl-1'-pyrophosphateÂ (PRPP) synthesis suggests that cells exposed to OA suffer PRPP deprivation. We hypothesized that the PRPP pools were depleted as a result of an increase in the synthesis of OMP stimulated by exogenous OA. PRPP depletion would result in a marked decrease in purine synthesis, which would lead to an imbalance between purine and pyrimidine nucleotides, causing arrest of DNA synthesisÂ and a lack of energetic nucleotides (ADP, ATP). Three independent biological materials with one dyeswap were performed.

ORGANISM(S): Rhizobium etli CFN 42

SUBMITTER: Sergio EncarnaciÃ³n

PROVIDER: E-GEOD-46013 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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How does orotic acid affect the growth of R.etli CE3?

Project description:Growth inhibition of Rhizobium etli and other organisms by exogenous OA has not been previously reported. We conducted genome-wide transcriptomic analysis to determine the possible mechanism by which OA exerts its growth-inhibitory effect on R. etli CE3. The observed changes fall into several broad categories. First, the decreased expression of genes involved in protein synthesis (including ribosomal genes), replication and energy production indicates that the cells arrest or at least slow their growth, which agrees with the observed growth arrest induced by OA. The upregulation of a key enzyme (PrsAch) for 5'-phosphoribosyl-1'-pyrophosphate (PRPP) synthesis suggests that cells exposed to OA suffer PRPP deprivation. We hypothesized that the PRPP pools were depleted as a result of an increase in the synthesis of OMP stimulated by exogenous OA. PRPP depletion would result in a marked decrease in purine synthesis, which would lead to an imbalance between purine and pyrimidine nucleotides, causing arrest of DNA synthesis and a lack of energetic nucleotides (ADP, ATP).

2013-10-12 | GSE46013 | GEO

Changes in liver proteins of rats fed both low and high-fat and sucrose diets induced by fish omega-3 PUFAs and their combination with grape polyphenols according to quantitative proteomics, part 2

Project description:This study considers the physiological modulation of liver proteins due to the supplementation with fish oils under two different dietary backgrounds: low- or high- fat and sucrose diets, and the effect of their combination with an antioxidant agent (grape polyphenols) which provides reducing power. For this scope, a quantitative proteomics approach based on the Isobaric Tag for relative and Absolute Quantitation methodology (iTRAQ)-coupled to nano-LC-MS/MS and complemented with 2D-DIGE analysis were used for determining the regulation of liver proteins exerted by the supplementation with fish oils, polyphenols or their combination of Wistar Kyoto rats in the two chosen dietary backgrounds. This experimental design was useful to investigate if the behavior of fish oils changes when the dietary background is modified and the possible synergy between fish oils and polyphenols. Results show that the capacity of fish oils, polyphenols or their combination for down or up-regulating liver proteins depends on the dietary context. In the background of low-fat low-sucrose healthy diets, 10 different proteins were altered by the sum of three supplements, in opposite to the 45 altered proteins found in the high-fat high-sucrose unhealthy diets. In both situations, fish oils seemed to be the main force for regulating liver proteins, although the addition of polyphenols was able to modulate some fish oils effects. Moreover, we provide evidence of the effect of fish oils and their combination with grape polyphenols for improving biochemical parameters and for reducing enzymes of hepatic lipogenesis and glycolysis, for enhancing fatty acid beta oxidation and insulin signaling and for the amelioration of endoplasmic reticule stress and protein oxidation when are included in an unhealthy diet.

2017-01-17 | PXD003080 | Pride

Potential Mechanism of Action of 3'-Demethoxy-6-O-Demethylisoguaiacin on Methicillin-resistant Staphylococcus aureus

Project description:Methicillin-resistant Staphylococcus aureus is one of the major causative agents associated to infections with a high morbidity and mortality in hospitals worldwide. In previous studies, we reported that lignan 3'-demethoxy-6-O-demethylisoguaiacin isolated and characterized from Larrea tridentata showed the best activity towards methicillin-resistant S. aureus. Understanding of mechanism of action of drugs allows design drugs in a better way. Therefore, we employed microarray to obtain gene expression profile of methicillin-resistant S. aureus after exposure to 3'-demethoxy-6-O-demethylisoguaiacin. The results showed that lignan had an effect on cell membrane affecting proteins of the ATP-binding cassette (ABC) transport system causing bacteria death. This study consisted of comparison of isolated RNA of MRSA not treated and MRSA treated with lignan 3'-demethoxy-6-O-demethylisoguaiacin. Both RNAs samples were differentially dyed with Cy3 and Cy5 during cDNA synthesis and hybridized on DNA chip. Afterwards, the chip was scanned in a GenePix 4000B scanner. The resulting gene expression profile was analyzed in databases for functional annotations to find a potential mechanism of the lignan in MRSA.

2015-06-27 | E-GEOD-70309 | biostudies-arrayexpress

Hormonal stimulation of carp is accompanied with changes of seminal plasma and spermatozoa proteome

Project description:Hormonal stimulation in common carp is a routine practice to enhance sperm production and control gamete maturation. The mechanism regulating functional changes of semen composition upon hormone treatment is unknown at present. The aim of the study was to compare proteome of carp spermatozoa and seminal plasma from control (PBS, n=10) and hormonally stimulated males (D-Ala6,Pro9-NEt)-mGnRH+metoclopramide, n=10). Comparative proteomics based on two-dimensional differential in-gel electrophoresis was employed to quantitatively analyze the spermatozoa and seminal plasma proteins changed in abundance due to hormonal stimulation. Changes in phosphorylation and oxidation level of spermatozoa proteins were also analyzed. The proteomic results were validated by western blot, enzymatic assay and mRNA expression analysis.In total we identified 62 spots (38 up and 24 down regulated) corresponding to 48 proteins with differential abundance in spermatozoa and 36 spots (23 up and 13 down regulated) corresponding to 23 proteins differentially abundant in seminal plasma upon hormone treatment (P<0.05; fold change 1.2). The majority of spermatozoa proteins changed in abundance as well as phosphorylation and carbonylation level were involved in cillium assembly and organization, energy metabolism, protein folding, and signal transduction while differentially abundant seminal plasma proteins were associated with immune and stress response, lipid metabolism and tissue remodelling. Our results strongly suggest that hormonal treatment is not only related to hydration of testis but also induces modification of spermatozoa proteins and synthesis of seminal plasma proteins. ROS signaling and cAMP/PKA and calcium/calmodulin pathways are involved in sperm response to hormonal stimulation.

2021-04-20 | PXD011049 | Pride

Mammary gland development: cross-species analysis of the mammary gland transcriptome in pregnant or lactating wild type female Sprague Dawley rats.

Project description:Cross-species hybridization analysis of mammary glands during pregnancy and lactation. Results provide insight into putative conserved molecular mechanisms regulating mammary gland development. This study was performed to identify orthologous transcripts that are differentially co-expressed in the mammary gland at 2 stages of development (pregnancy and lactation) in wild type Sprague-Dawley rats. Key points are examined in a time series of Sprague Dawley rat mammary gland development, secretory activation and lactation. Triplicate rat (three biological replicates) at each time point were used for statistical power totalling 12 individual arrays in this study. Rats were as staged pregnant day 1 the day that post coital plug was observed, and similarly, lactation day 1 was the first day after birth. Whole mammary glands No. 4 (inguinal) were obtained from female rats at stages of development: virgin (adulthood, 14 wks of age), Pregnant (5 and 14 days of pregnancy) and Lactating (day 1 and 12 postpartum). The two-color (Cy5/Cy3) microarray experiment was designed to hybridize samples from each group against a common reference, a pool of RNA from mammary gland of three parous or virgin female rats.

2013-10-30 | E-GEOD-22545 | biostudies-arrayexpress

Hepatic proteome analysis of Atlantic salmon (Salmo salar) after exposure to environmental concentrations of human pharmaceuticals

Project description:Pharmaceuticals are pseudo persistent aquatic pollutants with unknown effects at environmentally relevant concentrations. Atlantic salmon (Salmo salar) were exposed to Acetaminophen: 54.77 ± 34.67; Atenolol: 11.08 ± 7.98 and Carbamazepine: 7.85 ± 0.13 µg•L-1 for 5 days. After Acetaminophen treatment, 19 proteins were differently expressed, of which 11 were significant with respect to the control group (eight up-regulated and three down-regulated). After Atenolol treatment, 7 differently expressed proteins were obtained in comparison with the control, of which 6 could be identified (four up-regulated and 2 down-regulated). Carbamazepine exposure resulted in 15 differently expressed proteins compared with the control, with 10 of them identified (seven up-regulated and three down-regulated). Out of these, 3 features were common between Acetaminophen and Carbamazepine and one between Carbamazepine and Atenolol. One feature was common across all treatments. Principal component analysis and heat map clustering showed a clear grouping of the variability due to the applied treatments. The obtained data suggest (1) that exposure to environmentally relevant concentrations of the pharmaceuticals alters the hepatic protein expression profile of the Atlantic salmon; and (2) the existence of treatment specific processes that may be useful for biomarker development.

2014-11-18 | PXD001354 | Pride

H3K27ac ChIP-seq BTIC vs. DGC

Project description:Brain tumor initiating cells (BTICs), also known as cancer stem cells, hijack high-affinity glucose uptake normally active in brain to maintain energy demands in dynamic tumor microenvironments. Using unbiased metabolomics and genomic analyses, we discovered that de novo purine synthesis is functionally upregulated in BTICs, mediating glucose-sustained anabolic metabolism. Inhibiting purine synthesis abrogated BTIC growth, self-renewal, and in vivo tumor formation by depleting intracellular pools of purine nucleotides, supporting purine synthesis as potential therapeutic point of fragility. In contrast, differentiated brain tumor cells retained proliferative potential with targeting of purine biosynthetic enzymes, suggesting a selective dependence in BTICs. Upstream transcriptional activation of purine synthesis is mediated by MYC. Elevated expression of purine synthetic enzymes correlates with poor prognosis in glioblastoma patients. Collectively, our results suggest that a stem-like state in brain cancer is associated with metabolic reprogramming to fuel tumor hierarchy, revealing potential BTIC cancer dependencies amenable to targeted therapy.

2017-03-20 | GSE95216 | GEO

Identification of multiple transferrin species in spleen from mice with collagen-induced arthritis which may reflect changes in transferrin glycosylation associated with the disease activity. Role of CD38. Citrullinated Proteins

Project description:Collagen type II-induced arthritis (CIA) is an autoimmune disease that is accompanied by a complex host systemic response, which includes inflammatory and autoimmune reactions. Since to develop CIA is required the injection of antigen in complete Freund’s adjuvant (CFA), which is a water-in-mineral-oil emulsion containing killed Mycobacteria, systemic response proteins related with inflammation can confound the detection or diagnosis of arthritis. CIA in CD38 deficient mice (CD38 KO) is milder than that in C57BL/6 (B6 WT) mice. Protein extracts from spleen were subjected to 2D-DiGE and MS-MALDI-TOF/TOF analysis to identify proteins that were differentially expressed in CD38 KO and B6 WT mice with arthritis, with inflammation, or non-immunized. Differential protein expression was validated by ELISA, or Western-blotting.

2015-12-11 | PXD003183 | Pride

Distinct Serum Proteome Profiles Associated with Collageninduced Arthritis, and Complete Freund’s Adjuvant-induced Inflammation in CD38-/- mice: the Discriminative Power of Protein Species or Proteoforms

Project description:Collagen-type-II-induced arthritis (CIA) is an autoimmune disease, which involves a complex host systemic response including inflammatory and autoimmune reactions. CIA in C57BL/6 mice resembles human rheumatoid arthritis (RA) in terms of its disease course, histological findings, and in its response to commonly used anti-arthritic drugs. In this study, the goal has been to identify proteins from serum that change in their abundance in CD38-KO versus WT mice which may reflect their distinct response to an antigen-challenge that induces the development of an autoimmune disease. CIA is milder in CD38-/- than in Wild-type (WT) mice. We analyzed the sera from CD38-/- versus WT mice either with arthritis (CIA+), with no arthritis (CIA-), or with inflammation (Complete Freund’s adjuvant (CFA)-treated mice). To decrease the dynamic concentration range of serum a combinatorial ligand library composed of hexapeptides was used (called ProteoMiner). ProteoMiner-equalized serum samples were then subjected to 2D-DiGE and MS-MALDI-TOF/TOF analyses to identify proteins that changed in their relative abundances. Multivariate analyses revealed that a multi-protein signature (n = 28) was able to discriminate CIA+ from CIA- mice, and WT from CD38-/- mice within each condition. Likewise, a distinct multi-protein signature (n = 16) was identified which differentiated CIA+ CD38-/- mice from CIA+ WT mice, and lastly, a third multi-protein signature (n = 18) indicated that CD38-/- and WT mice could be segregated in response to CFA treatment This approach allows the identification of multiple protein species, or proteoforms of a given protein in a single analysis, and therefore, to focus the interest in fully characterize just the protein species that differ in abundance.

2015-07-17 | PXD001788 | Pride

Hypoxia induces Foxn1 and Mmp-9 expression in keratinocytes: proteomics analysis of mouse keratinocytes transiently activated by Foxn1

Project description:Recent studies have shown that transcription factor Foxn1 expressed in keratinocytes is involved in skin wound healing process, yet how Foxn1 functions remains largely unknown. Proteomic analysis using a two-dimensional differential gel electrophoresis (2D-DIGE) technique coupled with MALDI-TOF/TOF was used to analyze in vitro cultured keratinocytes transfected with adenoviral vector carrying Foxn1-GFP or GFP-alone (control). Sixty-six of sixty-nine protein spots differed in abundance between compared groups were identified.

2018-04-09 | PXD007843 | Pride

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