Endocannabinoid and lipid metabolism genes network analysis in adipose and liver tissue of dairy cows during the transition period

During the transition period dairy cows are exposed to enormous metabolic changes. These changes could affect the overall health and production. Two experiments were conducted on transition cows to evaluate the 1) effects of body condition score (BCS) on the endocannabinoid system and lipid metabolism gene expression in adipose tissue; and 2) effects of rumen-protected methionine on the endocannabinoid system in liver tissue. In the first study, cows were retrospectively classified according to their BCS at -3 wk from parturition into two groups: HiBCS (BCS ≥ 3.75) or LoBCS (BCS ≤ 3.25 ). Adipose tissue at -10, 7, and 20 d around parturition was used to examine mRNA expression via qPCR of endocannabinoid receptors (CNR1, CNR2), enzymes that synthesize endocannabinoid (NAPEPLD), enzymes that degrade endocannabinoids (FAAH, NAAA, MGLL), and the hormone precursor proopiomelanocortin (POMC). We also examined mRNA expression via qPCR of genes associated with lipolysis (LIPE, ABDH5, ATGL), fatty acid oxidation (CPT1A, CPT2, ACADVL, ACOX1), oxidative stress (SOD1, SOD2), and genes that are involved in inflammation (TLR9, TLR4, NFE2L2). Expression of CNR2 and NAPEPLD was greater at 7 d in LoBCS due to lower expression at the same time in HiBCS. The expression of FAAH was upregulated at d 7 and 20 in LoBCS than HiBCS cows. Expression of MGLL was overall greater across time in LoBCS than HiBCS, LoBCS had a tendency for greater overall expression of POMC across time. Regarding the genes associated with lipolysis, LoBCS compared with HiBCS cows had overall greater expression of ABDH5, LIPE and ATGL, indicating a greater state of basal lipolysis over time. Among genes related with fatty acid oxidation the expression of CPT1A and ACADVL was greater in HiBCS than LoBCS due to greater expression at -10 and 7 d. For the mitochondrial enzyme SOD2, important for clearing reactive-oxygen species that cause cellular stress and inflammation, we observed an interaction of BCS × day due to higher expression at d 7 in LoBCS than HiBCS. There was an overall BCS effect on the expression of SOD1 due to greater expression in LoBCS compared with HiBCS. In the second experiment, cows were fed experimental diets consisting of a basal control diet (CON) or rumen-protected methionine-supplemented (MET) during the transition period (-21 through 30 days in milk). The liver was biopsied at -10, 7, 20 and 30 days relative to parturition. Gene expression was determined through qPCR for endocannabinoid receptors (CNR1, CNR2), enzymes that synthesize endocannabinoid (NAPEPLD), enzymes that degrade endocannabinoid (FAAH, NAAA, MGLL), and the hormone precursor proopiomelanocortin (POMC). A significant interaction of treatment × day was observed for the endocannabinoid receptor CNR2 associated with lower expression in MET compared with control cows on d -10. There was an overall greater expression of FAAH, MGLL, NAAA and the EC-synthesizing enzyme NAPEPLD in MET compared with control cows. Cows supplemented with MET had greater in vitro blood neutrophil phagocytosis, neutrophil oxidative burst and monocyte oxidative burst. Results from experiment 1 indicate that expression of the endocannabinoid system and lipid metabolism genes in adipose tissue may be associated with BCS. A potential linkage between those pathways and risk of disorders postpartum remains to be determined. Results from experiment 2 suggest that the alterations in the hepatic EC signaling network in response to MET might be involved in the positive effect on performance and liver function

High prevalence of functional vitamin deficiencies in a psychogeriatric ward

Choline (Ch) is involved in relevant neurochemical processes. It is the precursor and metabolite of acetylcholine (ACh). It plays a role in single-carbon metabolism and is an essential component of different membrane phospholipids (PLs). These PLs are structural components of cell membranes, and involved in intraneuronal signal transduction. An increased ACh release was found after Ch treatment in rat corpus striatum slices. An in vivo proton magnetic resonance study has analyzed Ch ingestion effect. This work which represents the first non invasive study for exploring in vivo human brain neurochemistry showed the transfer of an oral Ch load in the brain of normal volunteers. These results were not confirmed by other in vivo studies. Cellular membranes breakdown is suggested as a feature of neurodegeneration in acute (stroke) and chronic (Alzheimer’s and vascular dementias) brain disorders. The effects of exogenous CCPLs on different brain areas were largely studied. Our group has assessed the influence of treatment with the CCPL, choline alphoscerate (GPC) on brain cholinergic neurotransmission markers in an animal model of brain vascular injury. A neuroprotective effect of GPC alone or in association with acetylcholinesterase inhibitor, galantamine was found. These results suggest that GPC could stimulate the expression of vesicular ACh transporter and Ch transporter primarily in areas involved in cognitive processes. These cholinergic markers could represent an appropriate mean to investigate brain cholinergic pathways. In the lack of novel therapeutic strategies, safe compounds developed since a long time such as the CCPLs could have still a place in pharmacotherapy and would merit to be investigated by new clinical studies

Management during the dry period and its effect on hepatic and adipose tissue molecular biomarkers of metabolism and health in grazing dairy cows

A successful transition into lactation determines optimum production, reproduction, and health. The peripartum period is characterized by an inflammatory state that, if not controlled, could be detrimental to the cow. The first experiment examined hepatic and adipose gene expression in response to injections of a non-steroidal anti-inflammatory compound (Carprofen) on 1, 3, and 5 d postpartum. Results indicated that after calving both tissues respond to inflammation signals, underscoring its role in the normal homeorhetic adaptations to lactation. The second experiment investigated the effect of prepartal nutrition and its interaction with BCS on hepatic and adipose tissue transcriptome, and the liver one-carbon metabolism and transulfuration pathway. Cows were randomly allocated to one of four groups in a 2 × 2 factorial arrangement: 4.0 or 5.0 BCS prepartum (10-point scale) and dietary energy at 75 or 125% of estimated requirements during the close-up. Tissue biopsies were harvested at -1, 1 and 4 wk relative to parturition. The greater number of hepatic differentially expressed genes in BCS4 cows in response to increased prepartum feed allowance (1071 vs 310, over the entire transition period) indicated a greater responsiveness to prepartum nutrition than optimally-conditioned cows. Thus, overfeeding in late-pregnancy should be limited to underconditioned cows, while cows with optimal BCS should be maintained on an energy-restricted diet. Adipose tissue mRNA and microRNA expression further confirmed this hypothesis, and indicated a relationship between the immune and metabolic response of the adipose tissue underscoring the existence of a “self-regulatory” mechanism. The extensive analysis of the hepatic one-carbon metabolism and related pathways highlighted fundamental differences in the metabolic progression of grazing cows compared to their higher-yield counterpart in TMR-based systems. Results also indicated a greater flux through these pathways in optimally conditioned cows feed restricted prepartum. The third experiment examined the effect of over-feeding in both close-up and far-off periods on the adipose tissue transcriptome. Far-off over-feeding is usually a standard practice in seasonal grazing systems as, compared with TMR-fed cows, cows are thinner at the end of lactation. Adipose expression data revealed how overfed cows in the far-off period had greater adipogenesis, consistent with their rapid gain in BCS following dry-off, but a lower body fat mobilization in early lactation. The results indicated that neither strategy negatively affected the adaptations to lactation. However, to ensure a favorable transition, cows should be subjected to a small feed restriction in the close-up period, irrespective of far-off nutrition. Overall, results indicated a beneficial involvement of the immune system in the adaptation to lactation, and the possibility to regulate this process through prepartal BCS and nutrition management. As a result of the three studies, New Zealand farmers, through DairyNZ (the industry organization that represents all New Zealand dairy producers), are now discouraged to apply prophylactic pharmacological intervention early postpartum, in favour of nutritional management during the dry period. Our recommendation is for cows to be properly managed in late lactation and early dry period to attain optimal condition (e.g, BCS 5) by close-up (3 wks from calving). Subsequently cows will benefit from a controlled feed restriction (75–90% of requirements). On the other hand, cows in less than optimal condition (e.g. BCS ≤ 4) should be fed to requirements or slightly overfed (110-120% of requirements) before calving. This is an easily implementable strategy based on pasture allocation capable of benefitting the farmer with a minimum cost

Antioxidants in Animal Feed

This includes 17 original research articles and focuses on the knowledge on the effects of dietary antioxidants on host health and performance of production animals, including livestock, poultry and fish. It provides various nutritional approaches to improve antioxidant capacity and benefit host health in animal production. Production animals exposed to air pollution, heavy metals, or other stressful conditions can experience oxidative stress that may suppress animal health, performance, and production, subsequently impacting economic feasibility; hence, maintaining and improving oxidative status, especially through an appropriate nutrition strategy, are essential for normal physiological processes in animals. Promising research results have revealed that the administration of natural or synthetic antioxidants in an animal’s diet may be an important strategy to mitigate the negative influence induced by oxidative stress conditions. The Special Issue has been conceived to set out the knowledge on the effects of dietary antioxidants on host health and performance of production animals. It provides various nutritional approaches, mainly including vitamins, plant extracts, trace elements, non-essential amino acids, etc., to improve antioxidant capacity and benefit host health in livestock, poultry and fish. This book will encourage more scientists to move forward on the path to increasing knowledge on the effect of natural or synthetic antioxidants on the growth and health of production animals

PPAR/RXR Regulation of Fatty Acid Metabolism and Fatty Acid ω-Hydroxylase (CYP4) Isozymes: Implications for Prevention of Lipotoxicity in Fatty Liver Disease

Fatty liver disease is a common lipid metabolism disorder influenced by the combination of individual genetic makeup, drug exposure, and life-style choices that are frequently associated with metabolic syndrome, which encompasses obesity, dyslipidemia, hypertension, hypertriglyceridemia, and insulin resistant diabetes. Common to obesity related dyslipidemia is the excessive storage of hepatic fatty acids (steatosis), due to a decrease in mitochondria β-oxidation with an increase in both peroxisomal β-oxidation, and microsomal ω-oxidation of fatty acids through peroxisome proliferator activated receptors (PPARs). How steatosis increases PPARα activated gene expression of fatty acid transport proteins, peroxisomal and mitochondrial fatty acid β-oxidation and ω-oxidation of fatty acids genes regardless of whether dietary fatty acids are polyunsaturated (PUFA), monounsaturated (MUFA), or saturated (SFA) may be determined by the interplay of PPARs and HNF4α with the fatty acid transport proteins L-FABP and ACBP. In hepatic steatosis and steatohepatitis, the ω-oxidation cytochrome P450 CYP4A gene expression is increased even with reduced hepatic levels of PPARα. Although numerous studies have suggested the role ethanol-inducible CYP2E1 in contributing to increased oxidative stress, Cyp2e1-null mice still develop steatohepatitis with a dramatic increase in CYP4A gene expression. This strongly implies that CYP4A fatty acid ω-hydroxylase P450s may play an important role in the development of steatohepatitis. In this review and tutorial, we briefly describe how fatty acids are partitioned by fatty acid transport proteins to either anabolic or catabolic pathways regulated by PPARs, and we explore how medium-chain fatty acid (MCFA) CYP4A and long-chain fatty acid (LCFA) CYP4Fω-hydroxylase genes are regulated in fatty liver. We finally propose a hypothesis that increased CYP4A expression with a decrease in CYP4F genes may promote the progression of steatosis to steatohepatitis

Traumatic brain injury-associated epigenetic changes and the risk for neurodegenerative diseases

Epidemiological studies have shown that traumatic brain injury (TBI) increases the risk for developing neurodegenerative diseases (NDs). However, molecular mechanisms that underlie this risk are largely unidentified. TBI triggers widespread epigenetic modifications. Similarly, NDs such as Alzheimer’s or Parkinson’s are associated with numerous epigenetic changes. Although epigenetic changes can persist after TBI, it is unresolved if these modifications increase the risk of later ND development and/or dementia. We briefly review TBI-related epigenetic changes, and point out putative feedback loops that might contribute to long-term persistence of some modifications. We then focus on evidence suggesting persistent TBI-associated epigenetic changes may contribute to pathological processes (e.g., neuroinflammation) which may facilitate the development of specific NDs – Alzheimer’s disease, Parkinson’s disease, or chronic traumatic encephalopathy. Finally, we discuss possible directions for TBI therapies that may help prevent or delay development of NDs

High-Yielding Dairy Cows

The milk industry is largely based on dairy cattle production. After decades of great advancements in genetics, nutrition, and management, today, one cow can reach unprecedented levels of milk production. New challenges have been posed to preserving the health and welfare of these domestic animals. “High-Yielding Dairy Cows” is a collection of scientific papers focusing on three main areas: metabolic diseases, reproduction diseases, and herd (heath) management in confined and pasture production systems. This book aggregates knowledge from a molecular level to a more holistic approach on disease prevention and management, giving the reader an accurate overview of the current state of the art of this topic. It intends to contribute to ensuring the supply of ethical and responsible animal protein for about eight billion of people

Characterising the role of a putative mammalian aspartate dehydrogenase in hepatic lipogenesis and secretion of very low density lipoproteins

Hepatic lipogenesis and the secretion of very low-density lipoproteins are severely perturbed in orotic-acid fed rats. Proteomic analysis of this rodent fatty liver identified a protein enriched in the microsomal fraction. The protein contains a domain of unknown function 108 (DUF 108) and is found to compose a complete protein in archaebacteria and mammalian species, but a single domain in Caenorhabditis elegans protein Q19527. The cDNA for human protein A6ND91 was cloned and stable rat hepatoma cell lines, expressing recombinant A6ND91-FLAG tagged protein were produced. Significant increases in de-novo lipogenesis and secretion of triglyceride-enriched lipoproteins were observed. This thesis considers A6ND91 protein in terms of structure and proposed function. Previous work has indicated that the homologous protein Thermatoga maritime 1643 (TM1643) possesses NAD+ or NADP+-dependent dehydrogenase activity towards L-Aspartate, producing NADH or NADPH plus iminoaspartate, a substrate that rapidly degrades into oxaloacetate (OAA) and ammonia. In this thesis, analyses of primary and predicted secondary structure of A6ND91 and TM1643 revealed that residues binding NAD+/NADP+ and the proposed active catalytic residue, Histidine198, are conserved. Northern-blot analysis identified A6ND91 is highly expressed in human liver, moderately in kidney, and at low levels in the brain. Western-blot analysis identified that A6ND91 is predominately localised to the cytosol, with minimal amounts in the microsomal fraction. Proteomic analysis revealed that A6ND91 contains a phosphorylated residue, Serine168. The kinase potentially responsible for phosphorylation of the residue is Glycogen Synthase Kinase 3 (GSK3), a well-characterised enzyme involved in regulation of glycogen metabolism and acetyl-CoA production. Microarray analyses indicate that over-expression of A6ND91 perturbs activity of several metabolic pathways involved in carboxylic acid metabolism, cholesterol synthesis and complement activation. Closer inspection of the perturbed metabolic pathways revealed an overlap through the substrates OAA/pyruvate/acetyl-CoA. Assays were performed to establish the NAD+/NADH and NADP+/NADPH redox state in cell lines expressing A6ND91. Results established that NAD+/NADH ratio was significantly reduced, suggesting that A6ND91 may operate as a dehydrogenase. Collectively, results suggest that L-Aspartate dehydrogenase activity of TM1643 is conserved in human A6ND91, playing a key role in hepatic lipid and energy metabolism through the production of OAA and its subsequent conversion to pyruvate/acetyl-CoA and cholesterol

Safety and Efficacy of Feed Additives in Animal Production

This book focuses on animal nutrition strategies using various feed additives to enhance nutrient digestibility and utilization, modulate the antioxidant capacity, mineral status, immunity, and gut and/or rumen microbiota in livestock. This results in improvements in their growth, performance and production parameters, as well as the quality of animal products