Dietary Interventions Reduce Traditional and Novel Cardiovascular Risk Markers by Altering the Gut Microbiome and Their Metabolites

Aims: The current study investigates the role of diet in mediating the gut microbiome-cardiovascular association which has not yet been explored in humans.Methods and Results: Using a two-arm dietary intervention study in healthy participants (N = 70), we assessed the effects of omega-3 and fibre supplementation on gut microbiome composition and short-chain fatty acid (SCFA) production. We then investigated how changes in gut microbiome composition correlated with changes in traditional cardiovascular risk factors (cholesterol, triglycerides, blood pressure), cytokines, and novel validated markers such as GlycA and ceramides, previously linked to CVD incidence and mortality. Both interventions resulted in significant drops in blood pressure, cholesterol, proinflammatory cytokines, GlycA and ceramides (all P < 0.05). Decreases in the atherogenic low-density lipoprotein triglyceride fraction, in total serum cholesterol were correlated with increases in butyric acid-production [β(SE) = −0.58 (0.06), P < 0.001; −0.53 (0.04), P < 0.001] and nominally associated with increases in some butyrogenic bacteria. Drops in GlycA were linked to increases in Bifidobacterium [β(SE) = −0.32 (0.04), P = 0.02] and other SCFAs including acetic acid [β(SE) = −0.28 (0.04), P = 0.02] and propionic acid [β(SE) = −0.3 (0.04), P = 0.02]. Additionally, we report for the first-time reductions in specific ceramide ratios that have been shown to predict CVD mortality and major adverse cardiovascular events such as d18:1/16:0, d18:0/24:0, and d18:1/24:1 which were associated with the reduction in the abundance in Colinsella and increases in Bifidobacteriuim and Coprococcus 3 and SCFAs (all P < 0.05).Conclusion: Overall, these findings support the potential of using simple dietary interventions to alter validated biomarkers linked to cardiovascular risk via the gut microbiome composition and its metabolic functions.

Alterations in placental long chain polyunsaturated fatty acid metabolism in human intrauterine growth restriction

Fatty acids (FA) are critical for fetal brain development and are transferred across the placenta by membrane-bound FA transport proteins (FATP), translocases (FAT/CD36), and cytosolic binding proteins (FABP). The cytosolic protein perilipin-2 aids in neutral lipid storage within lipid droplets. Decreased placental nutrient transport is believed to contribute to intrauterine growth restriction (IUGR); however, IUGR placental lipid transport and metabolism are poorly understood. We hypothesized that protein expression of FATPs, FABPs, and perilipin-2 in human placenta is decreased and placental lipid content and incorporation into lipid classes are reduced in IUGR. Placental tissue of idiopathic IUGR (n=25) and gestational age-matched, appropriately grown for gestational age (AGA) fetuses (n=19) was collected. We determined protein expression of FABP4 and perilipin-2 in placental homogenate and FATPs (2, 4, 6, CD36) in syncytiotrophoblast microvillous plasma membrane (MVM) by Western blot. Lipid droplet area (Oil Red O stain) and cellular FA content (GC/MS) were measured in chorionic villous tissue. MVM expression of FATP6 and CD36 was significantly increased in IUGR. The concentrations of seven n−6 and n−3 species long chain polyunsaturated FAs (LCPUFA) were significantly increased in the triglyceride fraction in IUGR vs AGA placenta. In summary, MVM FATP6 and CD36 protein expression is increased and LCPUFA are preferentially routed toward cellular storage in TG in the IUGR placenta, possibly to protect against oxidative stress associated with cellular FA accumulation. We speculate that these changes may be caused by impaired efflux of FA across the fetal-facing syncytiotrophoblast basal plasma membrane in IUGR placenta.

Isolation and Analysis of Phospholipids in Dairy Foods

The lipid fraction of milk is one of the most complex matrixes in foodstuffs due to the presence of a high number of moieties with different physical and chemical properties. Glycerolipids include glycerol and two fatty acids esterified in positions sn-1 and sn-2 with higher concentration of unsaturated fatty acids than in the triglyceride fraction of milk. Sphingolipids consist of a sphingoid base linked to a fatty acid across an amide bond. Their amphiphilic nature makes them suitable to be added into a variety of foods and recent investigations show that phospholipids, mainly phosphatidylserine and sphingomyelin, can exert antimicrobial, antiviral, and anticancer activities as well as positive effects in Alzheimer’s disease, stress, and memory decline. Polar lipids can be found as natural constituents in the membranes of all living organisms with soybean and eggs as the principal industrial sources, yet they have low contents in phosphatidylserine and sphingomyelin. Animal products are rich sources of these compounds but since there are legal restrictions to avoid transmission of prions, milk and dairy products are gaining interest as alternative sources. This review summarizes the analysis of polar lipids in dairy products including sample preparation (extraction and fractionation/isolation) and analysis by GC or HPLC and the latest research works using ELSD, CAD, and MS detectors.

Comparison of the antioxidant properties of emu oil with other avian oils

The antioxidant properties of emu oil were compared with oils derived from the fat of other avian species. We first examined their free radical scavenging activity against the 2,2-diphenyl-1-picryl hydracyl radical. The concentration of emu oil in the test solution that caused 50% neutralisation (IC50) was variable (24.5 ± 5.9 mg/mL, range 5.3–55.4 mg/mL), but similar to values obtained for other ratites (10.7 ± 5.9 mg/mL). In contrast, the IC50 values for duck and chicken oil were much higher (118.0 ± 8.1 mg/mL). The variability in the radical scavenging activity of emu oil preparations may reflect variations in the diets of the birds, the processing protocol and/or the storage age of the oil. We also evaluated some of the ratite oils for their inhibitory capacity on human erythrocyte membrane oxidation, by measuring the reduction of the thiobarbituric acid-reactive substance (TBAR) production. Emu oil had a greater effect in decreasing TBAR production than either the ostrich or rhea oil, suggesting that it offers more protection than the other ratite oils against oxidative damage. In conclusion, we demonstrated that emu oil has both antioxidant properties in vitro and a protective role against oxidative damage in a model biological membrane system. The antioxidant or radical scavenging properties of emu oil appear to be due to minor constituents in the non-triglyceride fraction of the oil, while its high ratio of unsaturated to saturated fatty acids (UFA : SFA) offers protection against oxidative damage.

Effect of fish-oil supplementation on levels of (n-3) poly-unsaturated fatty acids in the lipoprotein fractions of bovine plasma

Eight mid-lactation Holstein-Friesian dairy cows were used in an experiment to investigate whether poor transfer to milk of (n-3) long chain (≥C20) poly-unsaturated fatty acids (n-3 LCPUFA), from diets containing fish oil, is related to their mode of transport in plasma lipoproteins. The cows were split into two groups of four, each of which was housed with 24 other cows which did not provide blood data in this experiment. All cows received a basal diet ad libitum consisting of (kg/t dry matter) silage (539), grain distillers’ grains (148), rolled barley (248) and soya (65). All cows also received a mid-day meal of 2·76 kg per cow per day of molassed sugar-beet shreds. The control group (C) received no oil supplement but those in the treatment group (F) received 300 g per cow per day of fish oil absorbed onto the sugar-beet shreds. Fish oil inclusion progressively depressed milk fat concentration of the four cows from each group over the 3 weeks of the experiment so that levels were approximately 20 g/kg lower for cows in treatment F than for those in C (P < 0·05). Mean concentrations over all cows of the long-chain (≥C20, n-3) poly-unsaturated fatty acids in milk fat (g/100 g) were 0·12 and 0·54 for treatments C and F respectively, whilst corresponding output in milk (g/day) increased from 116 to 3·91, an average efficiency of transfer of 4·6%. The fatty acid compositions of the lipoproteins were measured in plasma samples from the four cows from each group after 3 weeks on the experimental diets The proportions (g/100 g fatty acids) of C20: 5 (n-3) in cholesteryl (CE) and phospholipid (PL) of high-density lipoprotein (HDL) in the plasma were respectively increased from 0·97 to 4·39 (P < 0·05) and from 1·15 to 4·74 (P < 0·01) by the fish-oil supplementation. For C22: 6 (n-3), the proportions in CE and PL of HDL respectively increased (P < 0·001) from 0·02 to 0·23 and from 0·16 to 2·18 as a result of the supplementation. The mean proportions (g/100 g fatty acids) of C20: 5 (n-3) and C22: 6 (n-3) in the triglyceride fraction of very low-density lipoprotein were 1·05 and 2·3 respectively and were not significantly altered by fish-oil supplementation (P > 0·05). It is suggested that the low transfer efficiency of (n-3) LCPUFA to the milk is partly due to their transport in the plasma largely as components of HDL lipids which are unavailable to the mammary gland.

Nutrient Consumption and Pigmentation of Deep and Surface Colonizing Sapstaining Fungi in Pinus contorta

Summary In this paper, we examined the ability of deep and surface staining fungi to utilize wood tissue nutrients. Fungal isolates were inoculated onto fresh billets and γ-sterilised sawnwood, both from Pinus contorta, and also onto defined nutrient media. The wood samples were assessed for host viability, fungal growth and nutrient status. The results indicated that the most aggressive sapstain species on fresh logs was Ceratocystis coerulescens, followed consecutively by Leptographium spp., Ophiostoma minus, O. piliferum, O. piceae, O. setosum, O. pluriannulatum and Aureobasidium pullulans. HPLC analysis of soluble sugars in fungal-infected wood indicated that mannose was the most depleted sugar, followed by glucose. Lipid analysis of infected wood indicated that Leptographium spp. and C. coerulescens greatly reduced the triglyceride fraction and that there was a wide spectrum of consumption of triglyceridederived fatty acids between the fungi. On defined media, the carbon source mannose led to the darkest pigmentation for all tested fungi. For C. coerulescens, the order of pigmentation intensity for the remaining tested carbon sources was reversed when compared to the other fungal species.

Unreliable use of standard muscle hydration value in obesity

Intramuscular water content is assumed to be constant in humans independent of their anthropometric characteristics. To verify whether this assumption is correct, intramuscular water, proteins, glycogen, and both total and intramyocytic triglycerides were measured in 51 samples of rectus abdominis muscle obtained from 16 lean and 35 overweight and obese subjects (body mass index cutoff 24.9 kg/m2). Data (referred to as wet tissue) were analyzed by means of a composition model at the cellular level of the skeletal muscle (SM). The average SM water content was 76.3 ± 3.3% in normal-weight individuals and 65.7 ± 5.8% in obese subjects ( P < 0.0001). Total triglycerides were 5.5 ± 2.3% in controls and 19.0 ± 7.0% in obese subjects ( P < 0.0001). The intramyocytic triglyceride fraction was also increased in obese subjects. The composition model provides an explanation for the negative correlation between total triglycerides and intramuscular water, and some of the model parameters were determined from the experimental data. In conclusion, although the hydration of fat-free SM mass may be unchanged in obese subjects, the hydration of in toto muscle mass decreases as its lipid content increases.

Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida

Cattle, pig and sheep oocytes isolated from healthy cumulus-oocyte complexes were pooled, within species, to provide samples of immature denuded oocytes with intact zona pellucida (n = 1000 per sample) for determination of fatty acid mass and composition in total lipid, constituent phospholipid and triglyceride. Acyl-containing lipid extracts, transmethylated in the presence of a reference penta-decaenoic acid (15:0), yielded fatty acid methyl esters which were analysed by gas chromatograph. Mean (+/- SEM) fatty acid content in samples of pig oocytes (161 +/- 18 micrograms per 1000 oocytes) was greater than that in cattle (63 +/- 6 micrograms; P < 0.01) and sheep oocytes (89 +/- 7 micrograms; P < 0.05). Of 24 fatty acids detected, palmitic (16:0; 25-35%, w/w), stearic (18:0; 14-16%) and oleic (18:1n-9; 22-26%) acids were most prominent in all three species. Saturated fatty acids (mean = 45-55%, w/w) were more abundant than mono- (27-34%) or polyunsaturates (11-21%). Fatty acids of the n-6 series, notably linoleic (18:2n-6; 5-8%, w/w) and arachidonic acid (20:4n-6; 1-3%), were the most abundant polyunsaturates. Phospholipid consistently accounted for a quarter of all fatty acids in the three species, but ruminant oocytes had a lower complement of polyunsaturates (14-19%, w/w) in this fraction than pig oocytes (34%, w/w) which, for example, had a three- to fourfold greater linoleic acid content. An estimated 74 ng of fatty acid was sequestered in the triglyceride fraction of individual pig oocytes compared with 23-25 ng in ruminant oocytes (P < 0.01). It is concluded that the greater fatty acid content of pig oocytes is primarily due to more abundant triglyceride reserves. Furthermore, this species-specific difference, and that in respect of polyunsaturated fatty acid reserves, may underlie the contrasting chilling, culture and cryopreservation sensitivities of embryos derived from pig and ruminant (cattle, sheep) oocytes.

Distribution of the total quantity of fatty acid double bonds in lipid components of the blood in children with diabetes mellitus

Children suffering from insulin-dependent diabetes mellitus (IDDM) were examined for unsaturation, that is, total quantity of double bonds in individual fractions of blood serum lipids was assessed. Lipid fractions were isolated by thin-layer chromatography. Unsaturation was assessed by ozonation method. The research was aimed at elucidation of biochemical criteria of the degree of metabolic disorders in children with IDDM and at development of methods for quantitative assessment of such disorders. Thirty-two inpatients with IDDM aged 7 to 15 were examined over time. Control group included 10 healthy age-matched children. Twenty children were examined during compensation stage (group 1), twelve during decompensation with ketoacidosis (group 2). Total unsaturation parameters were found reliably decreased in children with IDDM both during compensation and decompensation stages vs. those in healthy controls. A relative decrease of unsaturated fatty acids in the fraction of uneterified fatty acids (p0.05) was characteristic of group 2, with the content of their fragments in triglyceride fraction being the maximal (p0.05). The pattern of total unsaturation distribution in serum lipid fractions may serve as an indicator for metabolic disorders and decompensation in IDDM.