Dietary Precursors Research Articles

Abstract A97: Dietary nitrosamines, genetic variation in DNA repair and metabolism genes, and bladder cancer risk in the Los Angeles Bladder Cancer Study

Abstract Background: N-nitroso compounds (NOCs) can cause direct DNA damage and are known to induce a wide range of tumors, including tumors of the bladder in animal models. Humans are exogenously exposed to NOCs via diet, in particular processed meats, and cigarette smoke. However, NOCs, such as the carcinogen N-nitrosodimethylamine (NDMA), can also be formed endogenously from their dietary precursors: nitrates, nitrites and amines. The heme found in red meat is a potent stimulator of this endogenous formation. Our previous findings from the Los Angeles Bladder Cancer Study show that intake of meats with concurrent high amine and heme content, such as salami and liver, are associated with an increased risk of bladder cancer. Once NOCs are formed, they need to be activated metabolically by cytochrome P450 enzymes into electrophilic intermediates to exert a carcinogenic effect. These intermediates then react with DNA to form bulky adducts which, if not repaired, can lead to DNA instability and ultimately DNA damage. These reactive intermediates can also undergo metabolism by glutathione S-transferases (GSTs), which results in their detoxification. Multiple DNA repair pathways are available to correct NOC-induced damage. Methods: We examined the role of dietary sources of NOCs and NOC precursors in conjunction with variation in genes involved in NOC metabolism and DNA repair in the risk of bladder cancer using 355 bladder cancer cases and 409 controls from the Los Angeles Bladder Cancer Study, a population-based case-control study. We tested each of 627 SNPs in 27 genes involved in NOC metabolism and DNA repair for effect modification of exposure to different dietary sources of NOCs and NOC precursors. Multiple testing was accounted for using a global Bonferroni correction, across all SNPs combined, and also at the gene and pathway levels. Results: A polymorphism in the base excision repair gene hOGG1 (Ser326Cys; rs1052133) was shown to modify the effect of intake of high amine and heme content meats with a globally corrected p-value of 0.014 (crude p-value = 3.7 x 10-5). We found that the previously identified positive association between intake of high amine and heme content meat intake and bladder risk was restricted to individuals carrying one or more copies of the hOGG1 codon 326 Cys allele, which has been reported to reduce OGG1 protein activity. Conclusion: Results of this study build upon our previous findings that consumption of meats with high amine and heme content, such as salami and liver, are potential risk factors for bladder cancer. Our current finding of modification of this association by a DNA repair variant supports a role for endogenous dietary nitrosamines in bladder carcinogenesis. Citation Format: Chelsea E. Catsburg, Roman Corral, Juan Pablo Lewinger, Amit D. Joshi, Manuela Gago-Dominguez, Jian-Min Yuan, Victoria K. Cortessis, Malcolm C. Pike, Mariana C. Stern. Dietary nitrosamines, genetic variation in DNA repair and metabolism genes, and bladder cancer risk in the Los Angeles Bladder Cancer Study. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A97.

Exceptional lipid storage mode of the copepod Boeckella poopoensis in a pampean salt lake, Argentina

AB Aquatic Biology Contact the journal Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections AB 15:275-281 (2012) - DOI: https://doi.org/10.3354/ab00432 Exceptional lipid storage mode of the copepod Boeckella poopoensis in a pampean salt lake, Argentina Germán A. Kopprio1,4,*, Gerhard Kattner2, Martin Graeve2, R. Hugo Freije3, Rubén J. Lara1,4 1Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto Argentino de Oceanografía, Bahía Blanca B8000FWB, Argentina 2Ecological Chemistry, Alfred Wegener Institute for Polar and Marine Research, Bremerhaven 27570, Germany 3Environmental Chemistry, Universidad Nacional del Sur, Bahía Blanca B8000CPB, Argentina 4Wetland Dynamics, Leibniz Center for Tropical Marine Ecology, Bremen 28359, Germany *Email: gkopprio@criba.edu.ar ABSTRACT: The lipid biochemistry of zooplankton was investigated in Lake Chasicó, a pampean salt lake in Argentina. The lipid biomass of the zooplankton community was dominated by the calanoid copepod Boeckella poopoensis. The major storage lipids during winter were wax esters and triacylglycerols, which reached up to 59 and 37% of the total lipids, respectively. A striking feature of the zooplankton fatty acid composition was the extraordinarily high level of 18:4(n‑3) and 20:4(n-3) fatty acids, the highest ever reported for the latter in zooplankton. During winter, 20:4(n-3) accounted on average for 20% of the total fatty acids in the wax ester fraction and 7% in the triacylglycerols. The close relationship (r = 0.83, p < 0.001) between the 2 fatty acids implies the biosynthesis of 20:4(n-3) in B. poopoensis by chain elongation of 18:4(n-3), a dietary precursor and flagellate marker. The accumulation of 20:4(n-3) may be also partially related to B. poopoensis grazing on heterotrophic protozoa or non-flagellated chlorophytes, although this fatty acid was almost absent in the seston fraction. In summer, wax esters were slightly lower (45%), compensated by higher phospholipid levels. The 16:0 fatty alcohol moiety was predominant in the wax esters of all samples, corroborating the opportunistic feeding behavior of B. poopoensis. The high amounts of wax esters in zooplankton are typical of marine species, suggesting that the wax ester biosynthesis of B. poopoensis and the extraordinary fatty acid composition are adaptations to the unstable environmental conditions of salt lakes. KEY WORDS: Polyunsaturated fatty acids · Biosynthesis · Trophic markers · Wax esters · 20:4(n-3) Full text in pdf format PreviousNextCite this article as: Kopprio GA, Kattner G, Graeve M, Freije RH, Lara RJ (2012) Exceptional lipid storage mode of the copepod Boeckella poopoensis in a pampean salt lake, Argentina. Aquat Biol 15:275-281. https://doi.org/10.3354/ab00432 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AB Vol. 15, No. 3. Online publication date: June 06, 2012 Print ISSN: 1864-7782; Online ISSN: 1864-7790 Copyright © 2012 Inter-Research.

Sesamin modulation of lipid class and fatty acid profile in early juvenile teleost, Lates calcarifer, fed different dietary oils

Sesamin, a major sesame seed lignan, has diverse biological functions including the modulation of molecular actions in lipid metabolic pathways and reducing cholesterol levels. Vertebrates have different capacities to biosynthesize long-chain PUFA from dietary precursors and sesamin can enhance the biosynthesis of ALA to EPA and DHA in marine teleost. Early juvenile barramundi, Lates calcarifer, were fed for two weeks on diets rich in ALA or SDA derived from linseed or Echium plantagineum, respectively. Both diets contained phytosterols and less cholesterol compared with a standard fish oil-based diet. The growth rates were reduced in the animals receiving sesamin regardless of the dietary oil. However, the relative levels of n-3 LC-PUFA in total lipid, but not the phospholipid, increased in the whole body by up to 25% in animals fed on sesamin with ALA or SDA. Sesamin reduced the relative levels of triacylglycerols and increased polar lipid, and did not affect the relative composition of phospholipid subclasses or sterols. Sesamin is a potent modulator for LC-PUFA biosynthesis in animals, but probably will have more effective impact at advanced ages. By modulating certain lipid metabolic pathways, sesamin has probably disrupted the body growth and development of organs and tissues in early juvenile barramundi.

Effect of graded dietary fish oil supplementation on body composition and fillet fatty acid composition of pike (Esox luciusL.)

A feeding experiment was conducted on northern pike, Esox lucius L. (123.6±33.3 g initial body weight) applying graded dietary fish oil supplementation resulting three dietary fat levels (without supplementation: 6.2% fat and 11.7, 17.4% fat levels with supplementations) in a recirculation system. Feed consumption, feed efficiency and protein utilization of pike was not affected by the treatment. Whole body lipid content analysis showed that the composition of pike was significantly affected by the increasing level of fish oil supplementation, although no relationship was detected between the dietary and the fillet lipid content, as well as the protein content of fish bodies. High docosahexaenoic acid (DHA, C22:6n-3) proportions were found in the muscle lipids (groups fed fish oil supplementation), as compared to the dietary fatty acid compositions suggesting that with dietary fish oil supplementation the dietary precursors (mainly EPA) enable pike to convert long chain highly unsaturated fatty acids, esp...

Stable carbon isotope composition of perirenal adipose tissue fatty acids from Engadine sheep grazing either mountain or lowland pasture1

To provide further insights into ruminant lipid digestion and metabolism, and into cis-9,trans-11 18:2 synthesis, 12 growing Engadine lambs grazing either mountain pasture (2,250 m above sea level; n = 6) or lowland pasture (400 m above sea level; n = 6) were studied. Both pastures consisted exclusively of C(3) plants. Before the experiment, all animals grazed a common pasture for 6 wk. Grasses and perirenal adipose tissues of the sheep were analyzed for fatty acids by gas chromatography. Stable C-isotope ratios (δ(13)C values in ‰ vs. the Vienna Pee Dee Belemnite standard) were determined in the composite samples by elemental analysis-isotope ratio mass spectrometry. The δ(13)C of the individual fatty acids were measured by gas chromatography-combustion-isotope ratio mass spectrometry. The δ(13)C value of the entire mountain pasture grass was -27.5‰ (SD 0.31), whereas that of the lowland pasture grass was -30.0‰ (SD 0.07). This difference was reflected in the perirenal adipose tissues of the corresponding sheep (P < 0.05), even though the δ(13)C values were less in the animals than in the grass. The δ(13)C values for cis-9 16:1 and cis-9 18:1 in perirenal fat differed between mountain and lowland lambs (P < 0.05). The 16:0 in the adipose tissue was enriched in (13)C by 5‰ compared with the dietary 16:0, likely as a result of partly endogenous synthesis. The δ(13)C values of cis-9,trans-11 18:2 (cis-9,trans-11 CLA) in the adipose tissue were smaller than those of its dietary precursors, cis-9,cis-12 18:2 and cis-9,cis-12,cis-15 18:3; conversely, the δ(13)C values of trans-11 18:1 were not, suggesting that large proportions of perirenal cis-9,trans-11 18:2 were of endogenous origin and discrimination against (13)C occurred during Δ(9)-desaturation. The same discrimination was indicated by the isotopic shift between 16:0 and cis-9 16:1 in the mountain grazing group. Furthermore, the δ(13)C values of cis-9,trans-11 18:2 were smaller relative to the precursor fatty acids in the mountain lambs compared with the lowland group. This result suggests a reduced extent of biohydrogenation in lambs grazing on mountain grass in comparison with those grazing on lowland grass. This was supported by the smaller cis-9,trans-11 18:2 concentrations in total fatty acids found in the adipose tissues of the lowland lambs (P < 0.001). The results of this study demonstrate that natural differences between δ(13)C values of swards from different pastures and the adipose tissue fatty acids could be used as tracers in studies of lipid metabolism in ruminants.

A multi nutrient concept to enhance synapse formation and function: science behind a medical food for Alzheimer’s disease

Alzheimer’s Disease (AD) is the leading cause of dementia. Epidemiological studies suggest that AD is linked with poor status of nutrients including DHA, B-vitamins and the vitamins E and C. Ongoing neurodegeneration, particularly synaptic loss, leads to the classical clinical features of AD namely, memory impairment, language deterioration, and executive and visuospatial dysfunction. The main constituents of neural and synaptic membranes are phospholipids. Supplemenation of animals with three dietary precursors of phospholipids namely, DHA, uridine monophosphate and choline, results in increased levels of brain phospholipids, synaptic proteins, neurite outgrowth, dendritic spines formation (i.e. the anatomical precursors of new synapses) and an improvement in learning and memory. Other nutrients act as co-factors in the synthesis pathway of neuronal membranes. For example B-vitamins are involved in methylation processes, thereby enhancing the availability of choline as a synaptic membrane precursor. A multi-nutrient concept that includes these nutrients may improve membrane integrity, thereby influencing membrane-dependent processes such as receptor function and amyloid precursor protein (APP) processing, as shown by reduced amyloid production and amyloid β plaque burden, as well as toxicity. Together, these insights provided the basis for the development of a medical food for patients with AD, Souvenaid<sup>®<sup/>, containing a specific combination of nutrients (Fortasyn™ Connect) and designed to enhance synapse formation in AD. The effect of Souvenaid on memory and cognitive performance was recently assessed in a proof-of-concept study, SOUVENIR I, with 212 drug-naïve mild AD patients (MMSE 20-26). This proof-of-concept study demonstrated that oral nutritional supplementation with Souvenaid<sup>®<sup/> for 12 weeks improves memory in patients with mild AD. To confirm and extend these findings, we have designed and initiated three additional studies. Two of these studies will be completed in 2011; Souvenir II, a 24-week European study, with 259 drug-naïve mild AD patients (MMSE≥20) and S-Connect, another 24-week study, with 527 mild-tomoderate AD patients (MMSE 14-24) using AD medication conducted in the US. The third is the EU-funded LipiDiDiet study, a 24-month study, which will enrol 300 people with prodromal AD to assess the effect on memory performance.

Stable Carbon Isotope Composition of c9,t11‐Conjugated Linoleic Acid in Cow's Milk as Related to Dietary Fatty Acids

This study explores the potential use of stable carbon isotope ratios (δ(13)C) of single fatty acids (FA) as tracers for the transformation of FA from diet to milk, with focus on the metabolic origin of c9,t11-18:2. For this purpose, dairy cows were fed diets based exclusively on C(3) and C(4) plants. The FA in milk and feed were fractionated by silver-ion thin-layer chromatography and analyzed for their δ(13)C values. Mean δ(13)C values of FA from C(3) milk were lower compared to those from C(4) milk (-30.1‰ vs. -24.9‰, respectively). In both groups the most negative δ(13)C values of all FA analyzed were measured for c9,t11-18:2 (C(3) milk=-37.0±2.7‰; C(4) milk -31.4±1.4‰). Compared to the dietary precursors 18:2n-6 and 18:3n-3, no significant (13)C-depletion was measured in t11-18:1. This suggests that the δ(13)C-change in c9,t11-18:2 did not originate from the microbial biohydrogenation in the rumen, but most probably from endogenous desaturation of t11-18:1. It appears that the natural δ(13)C differences in some dietary FA are at least partly preserved in milk FA. Therefore, carbon isotope analyses of individual FA could be useful for studying metabolic transformation processes in ruminants.

Arginine Can Be Synthesized from Enteral Proline in Healthy Adult Humans

There is considerable controversy recently in identifying dietary precursors for arginine synthesis. We have previously shown in human neonates and piglets that proline is the sole dietary precursor for arginine synthesis. It is unclear in adult humans whether proline is a dietary precursor for arginine. We performed a multi-tracer stable isotope study in adults using 15N2-ureido arginine and 15N proline to elucidate synthesis of citrulline and arginine and determine whether proline is a precursor for arginine. Primed, intermittent infusions of the labeled amino acids were given enterally to 5 healthy men consuming a standardized milkshake diet. Blood was sampled during plateau enrichment between 1.5 and 3 h. Plasma enrichment occurred for both tracers, giving enteral turnover estimates of 93 μmol · kg-1 · h-1 for arginine and 154 μmol · kg-1 · h-1 for proline. Appearance of the label from proline in arginine and the intermediaries, ornithine and citrulline, was measured in all participants. The rate of synthesis of arginine from proline was 3.7 μmol · kg-1 · h-1, which is estimated to be ~40% of newly synthesized arginine. In this first study in adult humans using an enteral proline tracer, we have demonstrated synthesis of arginine from this dietary amino acid. Therefore, as in newborns, proline must now be considered a dietary precursor for arginine in healthy adults.

Up-regulated Desaturase and Elongase Gene Expression Promoted Accumulation of Polyunsaturated Fatty Acid (PUFA) but Not Long-Chain PUFA in Lates calcarifer, a Tropical Euryhaline Fish, Fed a Stearidonic Acid- and γ-Linoleic Acid-Enriched Diet

The limited activity of Δ6 fatty acid desaturase (FAD6) on α-linolenic (ALA, 18:3n-3) and linoleic (LA, 18:2n-6) acids in marine fish alters the long-chain (≥C(20)) polyunsaturated fatty acid (LC-PUFA) concentration in fish muscle and liver when vegetable oils replace fish oil (FO) in aquafeeds. Echium oil (EO), rich in stearidonic acid (SDA, 18:4n-3) and γ-linoleic acid (GLA, 18:3n-6), may enhance the biosynthesis of n-3 and n-6 LC-PUFA by bypassing the rate-limiting FAD6 step. Nutritional and environmental modulation of the mechanisms in LC-PUFA biosynthesis was examined in barramundi, Lates calcarifer , a tropical euryhaline fish. Juveniles were maintained in either freshwater or seawater and fed different dietary LC-PUFA precursors present in EO or rapeseed oil (RO) and compared with FO. After 8 weeks, growth of fish fed EO was slower compared to the FO and RO treatments. Irrespective of salinity, expression of the FAD6 and elongase was up-regulated in fish fed EO and RO diets, but did not lead to significant accumulation of LC-PUFA in the neutral lipid of fish tissues as occurred in the FO treatment. However, significant concentrations of eicosapentaenoic acid (EPA, 20:5n-3) and arachidonic acid (ARA, 20:4n-6), but not docosahexaenoic acid (DHA, 22:6n-3), appeared in liver and, to a lesser extent, in muscle of fish fed EO with marked increases in the phospholipid fraction. Fish in the EO treatment had higher EPA and ARA in their liver phospholipids than fish fed FO. Endogenous conversion of dietary precursors into neutral lipid LC-PUFA appears to be limited by factors other than the initial rate-limiting step. In contrast, phospholipid LC-PUFA had higher biosynthesis, or selective retention, in barramundi fed EO rather than RO.

Carotene and Novel Apocarotenoid Concentrations in Orange-Fleshed Cucumis melo Melons: Determinations of β-Carotene Bioaccessibility and Bioavailability

Muskmelons, both cantaloupe (Cucumis melo Reticulatus Group) and orange-fleshed honeydew (C. melo Inodorus Group), a cross between orange-fleshed cantaloupe and green-fleshed honeydew, are excellent sources of β-carotene. Although β-carotene from melon is an important dietary antioxidant and precursor of vitamin A, its bioaccessibility/bioavailability is unknown. We compared β-carotene concentrations from previously frozen orange-fleshed honeydew and cantaloupe melons grown under the same glasshouse conditions, and from freshly harvested field-grown, orange-fleshed honeydew melon to determine β-carotene bioaccessibility/bioavailability, concentrations of novel β-apocarotenals, and chromoplast structure of orange-fleshed honeydew melon. β-Carotene and β-apocarotenal concentrations were determined by HPLC and/or HPLC-MS, β-carotene bioaccessibility/bioavailability was determined by in vitro digestion and Caco-2 cell uptake, and chromoplast structure was determined by electron microscopy. The average β-carotene concentrations (μg/g dry weight) for the orange-fleshed honeydew and cantaloupe were 242.8 and 176.3 respectively. The average dry weights per gram of wet weight of orange-fleshed honeydew and cantaloupe were 0.094 g and 0.071 g, respectively. The bioaccessibility of field-grown orange-fleshed honeydew melons was determined to be 3.2 ± 0.3%, bioavailability in Caco-2 cells was about 11%, and chromoplast structure from orange-fleshed honeydew melons was globular (as opposed to crystalline) in nature. We detected β-apo-8'-, β-apo-10', β-apo-12'-, and β-apo-14'-carotenals and β-apo-13-carotenone in orange-fleshed melons (at a level of 1-2% of total β-carotene). Orange-fleshed honeydew melon fruit had higher amounts of β-carotene than cantaloupe. The bioaccessibility/bioavailability of β-carotene from orange-fleshed melons was comparable to that from carrot (Daucus carota).

The effect of polyunsaturated fatty acids on bone health

SummaryThe essential polyunsaturated fatty acids (PUFAs) are divided into two classes, n-3 (ω-3) and n-6 (ω-6) and their dietary precursors are α-linolenic (ALA) and linoleic acid (LA), respectively. PUFAs are precursors of a wide range of metabolites, for example eicosanoids like prostaglandins and leukotrienes, which play critical roles in the regulation of a variety of biological processes, including bone metabolism.A large body of evidence supports an effect of PUFA on bone metabolism which may be mediated by regulation of osteoblastogenesis and osteoclast activity, change of membrane function, decrease in inflammatory cytokines, such as interleukin-1 (IL-1), interleukin-6 (IL-6), and tumour necrosis factor alpha (TNF-α), modulation of peroxisome proliferators-activated receptor γ (PPARγ) and influence in NO secretion and NO synthase.Animal studies have shown that a higher dietary omega-3/omega-6 fatty acids ratio is associated with beneficial effects on bone health. Human studies conducted in elderly subjects suggest that omega-3 instead of omega-6 has a positive effect on bone metabolism. In spite of increasing evidence, studies conducted in humans do not allow us to draw a definitive conclusion on the usefulness of PUFAs in clinical practice.

Arginine Is Synthesized From Proline, Not Glutamate, in Enterally Fed Human Preterm Neonates

In neonatal mammals, arginine is synthesized in the enterocyte, with either proline or glutamate as the dietary precursor. We have shown several times in piglets that proline is the only precursor to arginine, although in vitro evidence supports glutamate in this role. Because of this uncertainty, we performed a multitracer stable isotope study to determine whether proline, glutamate, or both are dietary precursors for arginine in enterally fed human neonates. Labeled arginine (M + 2), proline (M + 1), and glutamate (M + 3) were given enterally to 15 stable, growing preterm infants (GA at birth 30-35 wk) at 1-3 wk postnatal age. Enrichment in urine of the tracer amino acids and the M + 1 and M + 3 isotopomers of arginine were measured by LC-tandem mass spectrometry to determine the contribution of proline and glutamate to arginine synthesis. Plateau enrichments of arginine and proline tracers were measurable in urine. Urinary glutamate enrichment was not detected. Conversion of proline to arginine was detected. However, the M + 3 isotopomer of arginine, which would have been synthesized from glutamate, was not detected. We conclude that, in contrast to the current consensus in the literature based on in vitro studies, proline is the major contributor to arginine synthesis in human preterm infants.

Replacing dietary fish oil with Echium oil enriched barramundi with C 18 PUFA rather than long-chain PUFA

Vegetable oils (VO) are sustainable sources for replacement of fish oil (FO) in aquafeeds. However, VO lacks the health-benefitting n-3 long-chain (≥ C 20) polyunsaturated fatty acids (n-3 LC-PUFA) and potentially compromise farmed fish flesh quality for consumers. In a factorial experiment, barramundi ( Lates calcarifer) were grown in either freshwater or seawater and fed on three diets containing different oil sources: FO; stearidonic acid (SDA, 18:4n-3) rich oil from Echium plantagineum (EO); or rapeseed oil (RO). RO and FO-fed fish grew faster than the EO treatment and all three dietary treatments were not affected by salinity. A fatty acid mass balance showed that feeding barramundi on EO diet bypassed the first rate-limiting step in n-3 LC-PUFA biosynthesis. However, the fish did not accumulate high EPA or DHA content. Total PUFA, mainly of the n-3 series and dominated by ALA (18:3n-3) and SDA, in the whole body of EO fish was higher than for the FO and RO treatments. The n-3:n-6 ratio in EO treatment was less than for FO, but exceeded that in RO-fed fish. FA apparent metabolism as derived from the fatty acid mass balance fluxes showed comparable kinetics for key enzymes, indicating limited efficiency for LC-PUFA biosynthesis from their C 18 dietary precursors in barramundi fed EO or RO containing diets. Fish digested dietary FA and accumulated them efficiently regardless of the salinity. These findings establish a more comprehensive understanding for FA metabolism in barramundi fed different dietary lipids and at extremes of the species wide salinity range. Based on the observed levels of accumulation, EO-fed barramundi are a potentially rich source of ALA and SDA for human consumption.

Ovariectomy and 17β-estradiol alter transcription of lipid metabolism genes and proportions of neo-formed n-3 and n-6 long-chain polyunsaturated fatty acids differently in brain and liver

Hormonal and nutritional factors regulate the metabolism of long-chain polyunsaturated fatty acids (LC-PUFA). We aimed to determine whether ovarian hormones influence the capacity of rats to synthesize the end-products 22:6n-3 (DHA) and 22:5n-6 (n-6DPA) from their respective dietary precursors (18:3n-3 and 18:2n-6), and can regulate PUFA conversion enzymes gene transcription in brain and/or liver. Females born with a low DHA status were fed from weaning to 8 weeks of age a diet providing both essential precursors, and were concurrently submitted to sham-operated control (SOC) or ovariectomy (OVX) in combination with or without 17β-estradiol (E2) dosed at 8 or 16 μg/day. Relative to SOC, OVX increased the hepatic Δ9-, Δ6- and Δ5-desaturase transcripts and cognate transcription factors (PPARα, PPARγ, RXRα, RARα), but it did not affect LC-PUFA contents in phospholipids. In comparison with SOC and OVX groups, both E2 doses prevented the increase of transcripts, while paradoxically augmenting DHA and n-6DPA in liver phospholipids. Thus, in the liver of rats undergoing ovariectomy, changes of LC-PUFA synthesizing enzyme transcripts and of LC-PUFA proportions were not correlated. In brain, ovariectomy did not modify the transcripts of lipid metabolism genes, but it decreased DHA (-15%) and n-6DPA (-28%). In comparison with SOC and OVX groups, ovariectomized females treated with E2 preserved their status of both LC-PUFA in brain and had increased transcripts of E2 receptor β, PPARδ, RARα and LC-PUFA synthesizing enzymes. In conclusion, E2 sustained the transcription of lipid metabolism genes and proportions of neo-formed DHA and n-6DPA differently in brain and liver.

Enteral arginase II provides ornithine for citrulline synthesis

The synthesis of citrulline from arginine in the small intestine depends on the provision of ornithine. To test the hypothesis that arginase II plays a central role in the supply of ornithine for citrulline synthesis, the contribution of dietary arginine, glutamine, and proline was determined by utilizing multitracer stable isotope protocols in arginase II knockout (AII(-/-)) and wild-type (WT) mice. The lack of arginase II resulted in a lower citrulline rate of appearance (121 vs. 137 μmol·kg(-1)·h(-1)) due to a reduced availability of ornithine; ornithine supplementation was able to restore the rate of citrulline production in AII(-/-) to levels comparable with WT mice. There were significant differences in the utilization of dietary citrulline precursors. The contribution of dietary arginine to the synthesis of citrulline was reduced from 45 to 10 μmol·kg(-1)·h(-1) due to the lack of arginase II. No enteral utilization of arginine was observed in AII(-/-) mice (WT = 25 μmol·kg(-1)·h(-1)), and the contribution of dietary arginine through plasma ornithine was reduced in the transgenic mice (20 vs. 13 μmol·kg(-1)·h(-1)). Dietary glutamine and proline utilization were greater in AII(-/-) than in WT mice (20 vs. 13 and 1.4 vs. 3.7 μmol·kg(-1)·h(-1), respectively). Most of the contribution of glutamine and proline was enteral rather than through plasma ornithine. The arginase isoform present in the small intestinal mucosa has the role of providing ornithine for citrulline synthesis. The lack of arginase II results in a greater contribution of plasma ornithine and dietary glutamine and proline to the synthesis of citrulline.

Metabolomics Study of Human Urinary Metabolome Modifications After Intake of Almond (Prunus dulcis (Mill.) D.A. Webb) Skin Polyphenols

Almond, as a part of the nut family, is an important source of biological compounds, and specifically, almond skins have been considered an important source of polyphenols, including flavan-3-ols and flavonols. Polyphenol metabolism may produce several classes of metabolites that could often be more biologically active than their dietary precursor and could also become a robust new biomarker of almond polyphenol intake. In order to study urinary metabolome modifications during the 24 h after a single dose of almond skin extract, 24 volunteers (n = 24), who followed a polyphenol-free diet for 48 h before and during the study, ingested a dietary supplement of almond skin phenolic compounds (n = 12) or a placebo (n = 12). Urine samples were collected before ((-2)-0 h) and after (0-2 h, 2-6 h, 6-10 h, and 10-24 h) the intake and were analyzed by liquid chromatography-mass spectrometry (LC-q-TOF) and multivariate statistical analysis (principal component analysis (PCA) and orthogonal projection to latent structures (OPLS)). Putative identification of relevant biomarkers revealed a total of 34 metabolites associated with the single dose of almond extract, including host and, in particular, microbiota metabolites. As far as we know, this is the first time that conjugates of hydroxyphenylvaleric, hydroxyphenylpropionic, and hydroxyphenylacetic acids have been identified in human samples after the consumption of flavan-3-ols through a metabolomic approach. The results showed that this non-targeted approach could provide new intake biomarkers, contributing to the development of the food metabolome as an important part of the human urinary metabolome.

Morphology, proliferation, and ribonucleic acid and fractional protein syntheses in the small intestinal mucosa of young goats fed soy protein-based diets with or without amino acid supplementation

The study was designed to examine whether feeding soy protein isolate as partial replacement of casein (CN) affects jejunal protein synthesis and whether effects may be ameliorated by supplementation of those AA known to be at lower concentrations in soy protein isolate than in CN. Goat kids (14 d) were fed comparable milk protein diets, in which 50% of the crude protein was CN (CAS), soy protein isolate (SPI), or soy protein isolate supplemented with AA (SPIA) for 43 d (n=8 per group). On d 42, plasma concentrations of protein, urea, and AA were measured before and after morning feeding. In the morning of d 43, [15N]RNA from yeast [13mg/kg of body weight (BW)] was given with the diet to measure the reutilization of dietary RNA precursors for mucosal RNA biosynthesis. Four hours later, an oral dose of l-[1-13C]leucine (180mg/kg of BW) was administered and blood samples were collected between −15 and +45min relative to tracer administration for analysis of plasma 13C α-ketoisocaproic acid and 13C recovery in blood CO2. Kids were killed 60min after the tracer application, and jejunal tissue was collected to determine mucosal morphology, cell proliferation, enzyme activities, RNA synthesis, and fractional protein synthesis rate. Plasma protein concentrations were higher in CAS than in SPI and SPIA. Plasma concentrations of Thr were higher in CAS than in SPI and SPIA, and those of Met were lower in SPI than in CAS and SPIA. In mid-jejunum, villus circumferences were higher in CAS than in SPI and SPIA, and villus height and villus height:crypt depth ratio were higher in CAS than in SPI. In mid-jejunum, mucosal protein concentrations were higher in CAS than in SPI and SPIA and mucosal activities of aminopeptidase N tended to be higher in CAS than in SPI, whereas activities of dipeptidyl peptidase IV tended to be lower in SPI than in SPIA. Activities of 5′ nucleotidase and xanthine oxidase were lower in CAS than in SPI. The 13C recovery in blood CO2 tended to be higher in SPI than in CAS. In mid-jejunum, 15N enrichment of RNA tended to be higher in CAS than in SPI, and 13C enrichment of protein-bound Leu was higher in SPI than in CAS. In mid-jejunum, the fractional protein synthesis rate tended to be higher in SPI than in CAS. Our results revealed changes in intestinal growth after soy protein feeding that were associated with effects on intestinal RNA and protein synthesis but that were not ameliorated by AA supplementation.

Plasma Arginine and Ornithine Are the Main Citrulline Precursors in Mice Infused with Arginine-Free Diets , ,

Dietary arginine is the main dietary precursor for citrulline synthesis, but it is not known if other precursors can compensate when arginine is absent in the diet. To address this question, the contributions of plasma and dietary precursors were determined by using multitracer protocols in conscious mice infused i.g. either an arginine-sufficient diet [Arg(+)] or an arginine-free diet [Arg(−)]. The plasma entry rate of citrulline and arginine did not differ between the 2 diet groups (156 ± 6 and 564 ± 30 μmol·kg−1·h−1, respectively); however, the entry rate of ornithine was greater in the mice fed the Arg(+) than the Arg(−) diet (332 ± 33 vs. 180 ± 16 μmol·kg−1·h−1). There was a greater utilization of plasma ornithine for the synthesis of citrulline (49 ± 4 vs. 36 ± 3 μmol·kg−1·h−1, 30 ± 3% vs. 24 ± 2% of citrulline entry rate) in the mice fed the Arg(−) diet than the Arg(+) diet. The utilization of plasma arginine did not differ between the 2 diet groups for citrulline synthesis, either through plasma ornithine (∼29 ± 3 μmol·kg−1·h−1) or at the site of citrulline synthesis (∼12 ± 3 μmol·kg−1·h−1). The contribution of dietary proline to the synthesis of citrulline was mainly at the site of citrulline production (17 ± 1 μmol·kg−1·h−1), rather than through plasma ornithine (5 ± 0.4 μmol·kg−1·h−1). Dietary glutamine was utilized only at the site of citrulline synthesis (4 ± 0.2 μmol·kg−1·h−1). Dietary glutamine and proline made a greater contribution to the synthesis of citrulline in mice fed the Arg(−) diet but remained minor sources for citrulline production. Plasma arginine and ornithine are able to support citrulline synthesis during arginine-free feeding.

The effects of dietary carotenoid intake on carotenoid accumulation in the retina of a wild bird, the house finch ( Carpodacus mexicanus)

Carotenoid pigments accumulate in the retinas of many animals, including humans, where they play an important role in visual health and performance. Recently, birds have emerged as a model system for studying the mechanisms and functions of carotenoid accumulation in the retina. However, these studies have been limited to a small number of domesticated species, and the effects of dietary carotenoid access on retinal carotenoid accumulation have not been investigated in any wild animal species. The purpose of our studies was to examine how variation in dietary carotenoid types and levels affect retinal accumulation in house finches ( Carpodacus mexicanus), a common and colorful North American songbird. We carried out three 8-week studies with wild-caught captive birds: (1) we tracked the rate of retinal carotenoid depletion, compared to other body tissues, on a very low-carotenoid diet, (2) we supplemented birds with two common dietary carotenoids (lutein + zeaxanthin) and measured the effect on retinal accumulation, and (3) we separately supplemented birds with high levels of zeaxanthin – an important dietary precursor for retinal carotenoids – or astaxanthin – a dominant retinal carotenoid not commonly found in the diet (i.e. a metabolic derivative). We found that carotenoids depleted slowly from the retina compared to other tissues, with a significant (∼50%) decline observed only after 8 weeks on a very low-carotenoid diet. Supplementation with lutein + zeaxanthin or zeaxanthin alone significantly increased only retinal galloxanthin and ε-carotene levels, while other carotenoid types in the retina remained unaffected. Concentrations of retinal astaxanthin were unaffected by direct dietary supplementation with astaxanthin. These results suggest highly specific mechanisms of retinal carotenoid metabolism and accumulation, as well as differential rates of turnover among retinal carotenoid types, all of which have important implications for visual health maintenance and interventions.

Arginine utilization for citrulline synthesis in arginase II knockout mice

The synthesis of citrulline (Cit) from arginine (Arg) in the small intestine depends on the activity of arginase II (ARG2). To test the hypothesis that Arg is the main dietary precursor for Cit synthesis, despite the lack of ARG2, tracer studies were conducted in WT and ARG2 ko conscious mice. WT mice had a higher rate of appearance (Ra) of Cit (130 vs. 118 μmol•kg−1 • h−1) than ARG2 ko mice, but no difference in Ra of ornithine (Orn; 278 μmol•kg−1 • h−1) was observed. There were significant differences in the utilization of dietary Cit precursors. The contribution of dietary Arg to the synthesis of Cit was reduced from 44 to 11 μmol•kg−1 • h−1 due to the lack of ARG2. No arginine was utilized at the site of Cit synthesis in ARG2 ko mice (WT = 26 μmol•kg−1 • h−1) and the contribution of Arg through Orn was reduced (18 vs. 14 μmol•kg−1 • h−1). Pro and Gln utilization increased in ARG2 ko mice from 5.7 to 8.1, and 0.7 to 1.9 μmol•kg−1• h−1, respectively. Most of the contribution of Pro and Gln was at the site of Cit synthesis, rather than through plasma Orn. Arg is still the main dietary precursor for Cit synthesis, despite the lack of ARG2 at the site of Cit synthesis. Arg is converted to Orn elsewhere in the body, presumably by action of ARG1, and then utilized for Cit synthesis. The contribution of dietary Gln and Pro increased in ARG2 ko mice, but these amino acids are minor sources for Cit synthesis. Support: USDA 2533771314 and NIH KO1 RR24173.