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International Journal of Molecular... Nov 2018This study aimed to characterize the long non-coding RNA (lncRNA) expression in the bovine mammary gland and to infer their functions in dietary response to 5% linseed...
This study aimed to characterize the long non-coding RNA (lncRNA) expression in the bovine mammary gland and to infer their functions in dietary response to 5% linseed oil (LSO) or 5% safflower oil (SFO). Twelve cows (six per treatment) in mid lactation were fed a control diet for 28 days followed by a treatment period (control diet supplemented with 5% LSO or 5% SFO) of 28 days. Mammary gland biopsies were collected from each animal on day-14 (D-14, control period), D+7 (early treatment period) and D+28 (late treatment period) and were subjected to RNA-Sequencing and subsequent bioinformatics analyses. Functional enrichment of lncRNA was performed via potential regulated target genes located within 50 kb flanking regions of lncRNAs and having expression correlation of >0.7 with mRNAs. A total of 4955 lncRNAs (325 known and 4630 novel) were identified which potentially targeted 59 and 494 genes in LSO and SFO treatments, respectively. Enrichments of target genes of lncRNAs indicated potential roles of lncRNAs in immune function, nucleic acid metabolism and cell membrane organization processes as well as involvement in Notch, cAMP and TGF-β signaling pathways. Thirty-two and 21 lncRNAs were differentially expressed (DE) in LSO and SFO treatments, respectively. Six genes (, , , , and ) were identified as potential target genes of six DE lncRNAs. In conclusion, this study has identified lncRNAs with potential roles in mammary gland functions and potential candidate genes and pathways via which lncRNAs might function in response to LSO and SFA.
Topics: Animals; Cattle; Dietary Supplements; Female; Gene Expression Profiling; Gene Expression Regulation; Gene Ontology; Linseed Oil; Mammary Glands, Animal; RNA, Long Noncoding; RNA, Messenger; Reproducibility of Results; Safflower Oil
PubMed: 30445766
DOI: 10.3390/ijms19113610 -
The Journal of Nutritional Biochemistry Jun 2016Glucagon like peptide-1 (GLP-1) and serotonin are both involved in food intake regulation. GLP-1 release is stimulated upon nutrient interaction with G-protein coupled... (Comparative Study)
Comparative Study
Glucagon like peptide-1 (GLP-1) and serotonin are both involved in food intake regulation. GLP-1 release is stimulated upon nutrient interaction with G-protein coupled receptors by enteroendocrine cells (EEC), whereas serotonin is released from enterochromaffin cells (ECC). The central hypothesis for the current study was that nutrient-induced GLP-1 release from EECs is modulated by serotonin through a process involving serotonin receptor interaction. This was studied by assessing the effects of serotonin reuptake inhibition by fluoxetine on nutrient-induced GLP-1, PYY and CCK release from isolated pig intestinal segments. Next, serotonin-induced GLP-1 release was studied in enteroendocrine STC-1 cells, where effects of serotonin receptor inhibition were studied using specific and non-specific antagonists. Casein (1% w/v), safflower oil (3.35% w/v), sucrose (50mM) and rebaudioside A (12.5mM) stimulated GLP-1 release from intestinal segments, whereas casein only stimulated PYY and CCK release. Combining nutrients with fluoxetine further increased nutrient-induced GLP-1, PYY and CCK release. Serotonin release from intestinal tissue segments was stimulated by casein and safflower oil while sucrose and rebaudioside A had no effect. The combination with fluoxetine (0.155μM) further enhanced casein and safflower oil induced-serotonin release. Exposure of ileal tissue segments to serotonin (30μM) stimulated GLP-1 release whereas it did not induce PYY and CCK release. Serotonin (30 and 100μM) also stimulated GLP-1 release from STC-1 cells, which was inhibited by the non-specific 5HT receptor antagonist asenapine (1 and 10μM). These data suggest that nutrient-induced GLP-1 release is modulated by serotonin through a receptor mediated process.
Topics: Animals; Caseins; Cell Line; Dibenzocycloheptenes; Dietary Sucrose; Diterpenes, Kaurane; Enterochromaffin Cells; Enteroendocrine Cells; Fluoxetine; Gene Expression Regulation; Glucagon-Like Peptide 1; Glucagon-Like Peptide-1 Receptor; Heterocyclic Compounds, 4 or More Rings; In Vitro Techniques; Male; Mice; Safflower Oil; Serotonin; Serotonin Antagonists; Selective Serotonin Reuptake Inhibitors; Sus scrofa; Sweetening Agents
PubMed: 27142747
DOI: 10.1016/j.jnutbio.2016.03.006 -
The American Journal of Clinical... May 2024Extracellular vesicles (EVs) are proposed to play a role in the development of cardiovascular diseases (CVDs) and are considered emerging markers of CVDs. n-3 PUFAs are... (Randomized Controlled Trial)
Randomized Controlled Trial
Dietary n-3 polyunsaturated fatty acids alter the number, fatty acid profile and coagulatory activity of circulating and platelet-derived extracellular vesicles: a randomized, controlled crossover trial.
BACKGROUND
Extracellular vesicles (EVs) are proposed to play a role in the development of cardiovascular diseases (CVDs) and are considered emerging markers of CVDs. n-3 PUFAs are abundant in oily fish and fish oil and are reported to reduce CVD risk, but there has been little research to date examining the effects of n-3 PUFAs on the generation and function of EVs.
OBJECTIVES
We aimed to investigate the effects of fish oil supplementation on the number, generation, and function of EVs in subjects with moderate risk of CVDs.
METHODS
A total of 40 participants with moderate risk of CVDs were supplemented with capsules containing either fish oil (1.9 g/d n-3 PUFAs) or control oil (high-oleic safflower oil) for 12 wk in a randomized, double-blind, placebo-controlled crossover intervention study. The effects of fish oil supplementation on conventional CVD and thrombogenic risk markers were measured, along with the number and fatty acid composition of circulating and platelet-derived EVs (PDEVs). PDEV proteome profiles were evaluated, and their impact on coagulation was assessed using assays including fibrin clot formation, thrombin generation, fibrinolysis, and ex vivo thrombus formation.
RESULTS
n-3 PUFAs decreased the numbers of circulating EVs by 27%, doubled their n-3 PUFA content, and reduced their capacity to support thrombin generation by >20% in subjects at moderate risk of CVDs. EVs derived from n-3 PUFA-enriched platelets in vitro also resulted in lower thrombin generation, but did not alter thrombus formation in a whole blood ex vivo assay.
CONCLUSIONS
Dietary n-3 PUFAs alter the number, composition, and function of EVs, reducing their coagulatory activity. This study provides clear evidence that EVs support thrombin generation and that this EV-dependent thrombin generation is reduced by n-3 PUFAs, which has implications for prevention and treatment of thrombosis.
CLINICAL TRIAL REGISTRY
This trial was registered at clinicaltrials.gov as NCT03203512.
Topics: Humans; Extracellular Vesicles; Fatty Acids, Omega-3; Male; Female; Cross-Over Studies; Middle Aged; Double-Blind Method; Blood Coagulation; Blood Platelets; Dietary Supplements; Cardiovascular Diseases; Adult; Fish Oils; Aged; Fatty Acids
PubMed: 38484976
DOI: 10.1016/j.ajcnut.2024.03.008 -
Methods in Molecular Biology (Clifton,... 2016Fundamentally, oil bodies are discrete storage organelles found in oilseeds, comprising a hydrophobic triacylglycerol core surrounded by a half-unit phospholipid...
Fundamentally, oil bodies are discrete storage organelles found in oilseeds, comprising a hydrophobic triacylglycerol core surrounded by a half-unit phospholipid membrane and an outer shell of specialized proteins known as oleosins. Oil bodies possess a number of attributes that were exploited by SemBioSys Genetics to isolate highly enriched fractions of oil bodies through liquid-liquid phase separation for a number of commercial applications. The current chapter provides a general guide for the isolation of oil bodies from Arabidopsis and/or safflower seed, from which protocols can be refined for different oilseed sources. For SemBioSys Genetic's recombinant technology, therapeutic proteins were covalently attached to oleosins or fused in-frame with ligands which bound oil bodies, facilitating their recovery to high levels of purity during "upstream processing" of transformed seed. Core to this technology was oil body isolation consisting of simple manipulation including homogenization of seeds to free the oil bodies, followed by the removal of insoluble fractions, and phase separation to recover the oil bodies. During oil body enrichment (an increase in oil body content concomitant with removal of impurities), a number of options and tips are provided to aid researchers in the manipulation and monitoring of these robust organelles.
Topics: Arabidopsis; Lipid Droplets; Liquid-Liquid Extraction; Plant Oils; Seeds
PubMed: 26614290
DOI: 10.1007/978-1-4939-3289-4_13 -
Journal of Equine Veterinary Science Jun 2022Greenhouse gases emission from livestock is the major concern for the ecosystem. Despite the lower contribution of non-ruminants towards greenhouse gas emission as...
Greenhouse gases emission from livestock is the major concern for the ecosystem. Despite the lower contribution of non-ruminants towards greenhouse gas emission as compared to the ruminants, the emission of methane (CH) gas from equines is expected to be increased in future due to its increasing population. Thus, it is essential to find or screen potential anti-methanogenic agent in a cost-effective and quicker manner. Considering this, the present investigation was aimed to analyze anti-methanogenic characteristic of bioactive compounds of safflower oil by targeting methanogenesis catalyzing enzyme (Methyl-coenzyme M reductase; MCR) via in silico tool. Initially, a total of 25 compounds associated with safflower oil were selected and their drug-likeness traits were predicted through Lipinski's rule of 5. Of 25 compounds, 9 compounds passed all the parameters of Lipinski's rule of five. These 9 ligands were further submitted for ADME traits analysis using Swiss ADME tool. Results revealed the absence of Lipinski's violation and approval of drug-likeness attributes of methyl tetradecanoate, 3-isopropyl-6-methylenecyclohex-1-ene, trans-2,4-decadienal, cis-6-nonenal, limonene, syringic acids, matairesinol, acacetin, and 2,5-octanedione. Molecular docking analysis was performed for analyzing the affinity between the selected 9 ligands and MCR receptor using FRED v3.2.0 from OpenEye Scientific Software and Discovery Studio client v16.1.0. Results showed maximum binding interaction of acacetin with MCR with the chemguass4 score of -13.35. Other ligands showed comparatively lower binding affinity in the order of matairesinol (-12.43) > methyl tetradecanoate (-9.25) > cis-6-nonenal (-7.88) > syringic acids (-7.73) > limonene (-7.18) > trans-2,4-decadienal (-7.07) > 3-isopropyl-6-methylenecyclohex-1-ene (-7.01) > 2,5-octanedione (-7.0.). In a nutshell, these identified compounds were observed as potential agents to reduce CH production from equines by targeting MCR. This in silico study emphasized the role of safflower-associated compounds in developing anti-methanogenic drug for equines in future.
Topics: Animals; Ecosystem; Euryarchaeota; Greenhouse Gases; Horses; Ligands; Limonene; Molecular Docking Simulation; Oxidoreductases; Safflower Oil
PubMed: 35346771
DOI: 10.1016/j.jevs.2022.103938 -
NMR in Biomedicine Feb 2022Longitudinal (T ) relaxation of triglyceride molecules and water is of interest for fat-water separation and fat quantification. A better understanding of T relaxation...
Longitudinal (T ) relaxation of triglyceride molecules and water is of interest for fat-water separation and fat quantification. A better understanding of T relaxation could benefit modeling for applications in fat quantification and relaxation mapping. This work investigated T relaxation of spectral resonances of triglyceride molecules and water in liquid fat-water mixtures and its dependence on the fat fraction. Dairy cream and a safflower oil emulsion were used. These were diluted with distilled water to produce a variety of fat mass fractions (4.4% to 35% in dairy cream and 6.3% to 52.3% in safflower oil emulsion). T was measured at room temperature at 3 T using an inversion recovery STimulated Echo Acquisition Mode (STEAM) MR spectroscopy method with a series of inversion times. T variations as a function of fat fraction were investigated for various resonances. A two-component model was developed to describe the relaxation in a fat-water mixture as a function of the fat fraction. The T of water and of all fat resonances studied in this work decreased as the fat fraction increased. The relative variation in T was different for each fat resonance. The T of the methylene resonance showed the least variation as a function of the fat fraction. The proposed two-component model closely fits the observed T variations. In conclusion, this work clarifies how the T of major and minor fat resonances and of the water resonance varies as a function of the fat fraction in fat-water mixtures. Knowledge of these variations could serve modeling, analysis of MRI measurements in fat-water mixtures, and phantom preparation.
Topics: Emulsions; Humans; Magnetic Resonance Spectroscopy; Sunflower Oil; Triglycerides; Water
PubMed: 34636097
DOI: 10.1002/nbm.4629 -
Protein Expression and Purification Oct 2017Fibroblast growth factor 10 (FGF10) is a member of the FGF superfamily. It exhibits diverse biological functions, and is extensively used for fundamental research and...
Fibroblast growth factor 10 (FGF10) is a member of the FGF superfamily. It exhibits diverse biological functions, and is extensively used for fundamental research and clinical applications involving hair growth, tissue repair, and burn wounds. Oil bodies, obtained from oil seeds, have been exploited for a variety of biotechnology applications. The use of oil bodies reduces purification steps and costs associated with the production of heterogonous proteins. Here, recombinant human FGF10 (rhFGF10) was expressed in safflower (Carthamus tinctorius L.) seeds using oilbody-oleosin technology. A plant expression vector, pOTBar-oleosin-rhFGF10, was constructed and introduced into safflower using Agrobacterium tumefaciens transformation, and mature safflower plants were obtained by grafting. Oleosin-rhFGF10 was successfully transformed and expressed in safflower seeds and inherited to the T generation. Moreover, MTT assays demonstrated that oil bodies expressed oleosin-FGF10 had a dose-dependent effect on cellular proliferation. In conclusion, this may provide a method of producing oleosin-rhFGF10, and help us meet the increasing pharmacological demands for the protein.
Topics: Agrobacterium tumefaciens; Animals; B-Lymphocytes; Carthamus tinctorius; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Fibroblast Growth Factor 10; Gene Expression; Genetic Vectors; Humans; Lipid Droplets; Mice; Plant Proteins; Plants, Genetically Modified; Recombinant Proteins; Seeds
PubMed: 26384708
DOI: 10.1016/j.pep.2015.09.016 -
Journal of Dairy Science Sep 2022Ruminal biohydrogenation (BH) of unsaturated fatty acids (FA) reduces absorption of essential FA and can result in formation of bioactive FA that cause milk fat...
Ruminal biohydrogenation (BH) of unsaturated fatty acids (FA) reduces absorption of essential FA and can result in formation of bioactive FA that cause milk fat depression. Rates of biohydrogenation of unsaturated FA are commonly observed using in vitro systems and are not well described in vivo. Seven ruminally cannulated cows were enrolled in a 3 × 3 Latin square design study to quantify biohydrogenation of 18:1n-9, 18:2n-6, and 18:3n-3 using a recently developed in vivo BH assay. All cows were fed a common high corn silage basal diet. Biohydrogenation was quantified using a perturbation model that consisted of a bolus dose of 200 g of an oil enriched in each unsaturated FA (oleic acid, OA = 87% 18:1n-9 sunflower oil; linoleic acid, LA = 70% 18:2n-6 safflower oil; and α-linolenic acid, ALA = 54% 18:3n-3 flaxseed oil) and 12 g of 17:0 as a marker of rumen outflow. Rumen contents were sampled before and after the bolus and enrichment of the bolused FA modeled. Using first-order kinetics to model FA disappearance, the fractional rates of disappearance of 18:1n-9 was 0.597 per hour, 18:2n-6 was 0.618 per hour, and 18:3n-3 was 0.834 per hour, similar to rates previously reported with this approach. Rumen turnover of 17:0 was 0.123 per hour, 0.065 per hour, and 0.106 per hour during the OA, LA, and ALA treatments, respectively. The extents of BH were calculated to be 82.8, 90.4, and 88.6% for 18:1n-9, 18:2n-6, and 18:3n-3, respectively. Finally, compartmental modeling was used to quantify the amount of each unsaturated FA metabolized through trans-10 and trans-11 BH pathways. The recently developed in vivo BH assay was able to predict rates of BH and provide insight into rumen metabolism of individual FA and may be useful to future investigations.
Topics: Animals; Cattle; Diet; Fatty Acids; Fatty Acids, Unsaturated; Female; Hydrogenation; Lactation; Milk; Rumen; Silage; alpha-Linolenic Acid
PubMed: 35931484
DOI: 10.3168/jds.2022-21831 -
Molecules (Basel, Switzerland) Apr 2022Safflower seed oil (SSO) is considered to be an excellent edible oil since it contains abundant essential unsaturated fatty acids and lipid concomitants. However, the...
Safflower seed oil (SSO) is considered to be an excellent edible oil since it contains abundant essential unsaturated fatty acids and lipid concomitants. However, the traditional alkali-refined deacidification process of SSO results in a serious loss of bioactive components of the oil and also yields massive amounts of wastewater. In this study, SSO was first extracted by ultrasonic-assisted ethanol extraction (UAEE), and the extraction process was optimized using random centroid optimization. By exploring the effects of ethanol concentration, solid−liquid ratio, ultrasonic time, and the number of deacidification times, the optimum conditions for the deacidification of safflower seed oil were obtained as follows: ethanol concentration 100%, solid−liquid ratio 1:4, ultrasonic time 29 min, and number of deacidification cycles (×2). The deacidification rate was 97.13% ± 0.70%, better than alkali-refining (72.16% ± 0.13%). The values of acid, peroxide, anisidine and total oxidation of UAEE-deacidified SSO were significantly lower than those of alkali-deacidified SSO (p < 0.05). The contents of the main lipid concomitants such as tocopherols, polyphenols, and phytosterols in UAEE-decidified SSO were significantly higher than those of the latter (p < 0.05). For instance, the DPPH radical scavenging capacity of UAEE-processed SSO was significantly higher than that of alkali refining (p < 0.05). The Pearson bivariate correlation analysis before and after the deacidification process demonstrated that the three main lipid concomitants in SSO were negatively correlated with the index of peroxide, anisidine, and total oxidation values. The purpose of this study was to provide an alternative method for the deacidification of SSO that can effectively remove free fatty acids while maintaining the nutritional characteristics, physicochemical properties, and antioxidant capacity of SSO.
Topics: Alkalies; Carthamus tinctorius; Ethanol; Peroxides; Plant Oils; Safflower Oil; Technology; Ultrasonics
PubMed: 35408704
DOI: 10.3390/molecules27072305 -
Prostaglandins, Leukotrienes, and... Jan 2018Polyunsaturated fatty acids (PUFA) have important signalling roles in the hypothalamus, a region of the brain that regulates whole-body energy homeostasis. While...
Polyunsaturated fatty acids (PUFA) have important signalling roles in the hypothalamus, a region of the brain that regulates whole-body energy homeostasis. While evidence suggests that high PUFA intake can impact hypothalamic activity, the underlying molecular mechanisms regulated by essential dietary n-6 and n-3 PUFA (i.e., linoleic acid and α-linolenic acid, respectively) remain poorly described in this brain region. To differentiate the roles of essential dietary PUFA on hypothalamic function, we fed male rats high-fat diets (35% kcal/d) containing either safflower (linoleic acid) or flaxseed (α-linolenic acid) oil for 2 months. Control rats were fed a low-fat (16% kcal/d) diet containing soybean oil. Hypothalamic fatty acids and gene expression were investigated by gas chromatography and microarray, respectively. Safflower-fed rats had higher total n-6 PUFA content due to increases in linoleic acid, arachidonic acid, and osbond acid compared to the other diet groups, while flaxseed-fed rats had higher total n-3 content due to increases in α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, and docosahexaenoic acid. Safflower-fed rats showed augmented expression of genes related to hypothalamic insulin signalling compared to controls. This was mirrored by significant increases in phosphorylated AKT and AKT levels; indicative of increased PI(3)K/AKT pathway activity. These changes were not observed in the hypothalamus of flaxseed-fed rats. Our findings provide new molecular insights into how essential fatty acids influence the hypothalamus and, potentially, whole-body energy homeostasis. This work also provides new knowledge to better understand the impact of essential fatty acids on metabolic and behavioral phenotypes.
Topics: Animals; Diet, High-Fat; Fatty Acids; Fatty Acids, Omega-3; Fatty Acids, Omega-6; Gene Expression Regulation; Hypothalamus; Insulin; Linseed Oil; Male; Rats, Sprague-Dawley; Safflower Oil
PubMed: 29413363
DOI: 10.1016/j.plefa.2017.12.002