Concentrations Of Individual Fatty Acids Research Articles

Complete characterization of the fatty acid profile of mare milk from commercial semi-extensive farms during lactation

The present study aimed to thoroughly describe the profile of fatty acids in mare milk from farms with different management systems and changes during lactation. Eighteen mares belonging to three commercial farms (6 mares per farm) were milked during the complete lactation period (six months). Fat content was determined by infrared spectroscopy, while fatty acid methyl esters were analyzed using a GC-FID. Mare milk, in general, contained 47.5 % saturated, 25.3 % monounsaturated, and 26.5 % polyunsaturated fatty acids. A high proportion of 18:2n-6 and 18:3n-3 but a low concentration of their long-chain metabolites was observed, and seven branched-chain fatty acids were identified. Stage of lactation significantly influenced the content of most individual fatty acids, and in general, milk from mares managed under pasture-based feeding systems showed a higher content of n-3 polyunsaturated fatty acids. This study provides new insights into mare milk FA composition and into dynamics of mare milk nutritional quality as affected by lactation stage and grazing intensity.

Effects of paternal arachidonic acid supplementation on offspring behavior and hypothalamus inflammation markers in the mouse.

Arachidonic acid (AA) is involved in inflammation and plays a role in growth and brain development in infants. We previously showed that exposure of mouse sires to AA for three consecutive generations induces a cumulative change in fatty acid (FA) involved in inflammation and an increase in body and liver weight in the offspring. Here, we tested the hypothesis that paternal AA exposure changes the progeny's behavioral response to a proinflammatory insult, and asked whether tissue-specific FA are associated with that response. Male BALB/c mice were supplemented daily with three doses of AA for 10 days and crossed to non-supplemented females (n = 3/dose). Two-month-old unsupplemented male and female offspring (n = 6/paternal AA dose) were exposed to Gram-negative bacteria-derived lipopolysaccharides (LPS) or saline control two hours prior to open field test (OFT) behavioral analysis and subsequent sacrifice. We probed for significant effects of paternal AA exposure on: OFT behaviors; individual FA content of blood, hypothalamus and hypothalamus-free brain; hypothalamic expression profile of genes related to inflammation (Tnfa, Il1b, Cox1, Cox2) and FA synthesis (Scd1, Elovl6). All parameters were affected by paternal AA supplementation in a sex-specific manner. Paternal AA primed the progeny for behavior associated with increased anxiety, with a marked sex dimorphism: high AA doses acted as surrogate of LPS in males, realigning a number of OFT behaviors that in females were differential between saline and LPS groups. Progeny hypothalamic Scd1, a FA metabolism enzyme with documented pro-inflammatory activity, showed a similar pattern of differential expression between saline and LPS groups at high paternal AA dose in females, that was blunted in males. Progeny FA generally were not affected by LPS, but displayed non-linear associations with paternal AA doses. In conclusion, we document that paternal exposure to AA exerts long-term behavioral and biochemical effects in the progeny in a sex-specific manner.

Roughage quality determines the production performance of post-weaned Hu sheep via altering ruminal fermentation, morphology, microbiota, and the global methylome landscape of the rumen wall

Roughage quality is a crucial factor influencing the growth performance and feeding cost of ruminants; however, a systematic investigation of the mechanisms underlying this is still lacking. In this study, we examined the growth performance, meat quality, ruminal fermentation parameters, rumen microbiome, and tissue methylomes of post-weaned Hu sheep fed low- or high-quality forage-based diets. Our results showed that sheep in the alfalfa hay (AG) and peanut vine (PG) groups exhibited better growth performance, slaughter performance, and meat quality than sheep in the wheat straw group (WG). The sheep in the AG possessed relatively higher contents of serum immunoglobins (IgA, IgG, and IgM) and lower contents of serum inflammation factors (TNF-α, IL-1β, IL-6, and IL-8) than those in the WG and the PG did. In addition, the levels of blood T lymphocytes (CD4+ and CD8+) and the CD4-to-CD8 ratio were significantly higher in the AG sheep than in the WG sheep and PG sheep. The concentration of ruminal NH3-N was highest in WG sheep, whereas the concentrations of individual and total short-chain fatty acids (SCFAs) were highest in the PG sheep. The length, width, and surface area of ruminal papillae were markedly different among the three groups, with the sheep in the PG being the most morphologically developed. The main ruminal microbes at the genus level include Prevotella 1, Rikenellaceae RC9 gut group, norank f F082, Ruminococcus 1, and Ruminococcus 2. The relative abundances of certain species are positively or negatively associated with fermentation parameters and growth index. For example, the fibrolytic bacteria Ruminococcaceae UGG-001 showed positive relationships with the concentration of SCFAs, except propionate. In addition, the relative abundances of fibrolytic bacteria (e.g., Ruminoccus 1) showed a negative relationship with starch-degrading bacteria (e.g., Prevotellaceae). The genome-wide DNA methylation analysis revealed that rumen tissues in the PG sheep and WG sheep occupied different global DNA methylomes. The genes with differentially methylated promoters were involved in known pathways (e.g., the FoxO signaling pathway) and the Gene Ontology (GO) terms (e.g., anatomical structure morphogenesis) pertaining to rumen development. Two candidate genes (ACADL and ENSOARG00020014533) with hyper- and hypo-methylated promoters were screened as potential regulators of rumen development. In conclusion, roughage quality determines sheep growth performance via directly influencing rumen fermentation and microbiome composition, and indirectly affecting rumen development at the epigenetic level.

Influence of Krill Meal on the Performance of Post-Smolt Atlantic Salmon That Are Fed Plant-Based and Animal-Based Fishmeal and Fish Oil-Free Diets

The purpose of this study was to determine the influence of krill meal (KM) inclusion at various levels (0%, 2.5%, 5%) in plant-based and animal-based feeds, that were fishmeal (FM) and fish oil (FO) free, on Atlantic salmon growth. A FM/FO feed containing 0% KM was the control. Using a 2 × 3 factorial approach, diets were randomly assigned to one of 28 0.5 m3 flow-through tanks (n = 4 tanks per diet) initially stocked with 60 fish (148.4 ± 12.9 g; 23.6 ± 0.8 cm; condition factor (K) = 1.16 ± 0.08) each. Salmon were fed for 90 days using automatic feeders ad libitum. On day 45, stocking densities were reduced to 45 fish per tank by the random removal of 15 individuals to remove any potential of density affecting growth through the trial end. Water temperature, oxygen saturation, pH, and salinity throughout the trial were 11.8 °C, 103.5%, 7.38, and 32.0 g L−1, respectively. Fish fed plant-based feed without KM were lighter (p < 0.05) than all other groups at day 45 and 90, but those fed a plant-based feed with KM had comparable growth and feed intake compared to that of fish fed the control diet. Irrespective of the presence of KM, animal-based feeds achieved comparable weight growth (p > 0.05) to the control and 5% KM plant-based groups, with KM increasing feed intake (p < 0.05). Between day 45 and 90, feed conversion ratios increased in all groups except the control and 0% KM plant-based group, while specific growth rates (SGRs) decreased for all except the 0% KM plant-based diet. Between-group differences (p < 0.05) were also noted for the thermal growth coefficient. No differences were recorded in visceral or intestinal weight, and whole-body lipid levels were identical, proportional for all groups. Although differences (p < 0.05) were apparent in the concentrations of individual fillet fatty acids between groups, a 75 g serving size of any treatment would be sufficient to exceed daily intake recommendations for EPA + DHA. This trial determined that benefit, in terms of feed intake and growth performance, was gained when KM was added to plant-based feeds. However, no such advantage was observed when KM was used with animal-based feeds.

Associations of Fatty Acid Profile with Abdominal Obesity in Men.

The content of individual unsaturated fatty acids in blood plasma (measured by HPLC) and their association with abdominal obesity in a group of men (mean age 52.2 years) was analyzed. The abdominal obesity was diagnosed according to the criteria of the All-Russian Scientific Society of Cardiology (waist circumference >94 cm). Men with abdominal obesity had higher levels of ω-6 γ-linolenic and dihomo-γ-linolenic acids, as well as ω-3 eicosapentaenoic and docosahexaenoic acids. They also had significantly elevated plasma levels of triglycerides and glucose and lower levels of HDL. Using single-factor ROC analysis we determined optimal cut-off thresholds for fatty acid levels indicating the presence of abdominal obesity. The results of regression analysis showed that the level of γ-linolenic acid is directly associated with the chance of abdominal obesity.s.

Effect of genotype on chemical composition and fatty acid profile of guinea pig carcass (Cavia porcellus L.)

The aim of this study was to evaluate the effect of the genotype on the deposition curve of the chemical components and fatty acid profile of carcass of guinea pigs of Peru and Cieneguilla genotypes. Forty-eight male guinea pigs (24 per genotype), randomly distributed in pens with three animals each per genotype were used. Management and feeding protocols, up to 32 wk of age, were similar for both genotypes. The deposition curve of the chemical components was determined using the Gompertz equation. Data of the fatty acid profile were submitted to analysis of varianza under a Randomized Complete Block Design using the SAS Studio Environment software, with a significance level of α = 0.05. The results showed that the asymptote of the moisture and protein content in the carcass of the Peru genotype was higher (P < 0.05) than that of Cieneguilla genotype, but not in the fat content, which was similar (P > 0.05) in both genotypes. Likewise, the function of the relationship between the maximum deposition rate of the three chemical components and the content at adulthood (k) was similar (P > 0.05) in the two genotypes. The age of maximum moisture deposition rate and total protein at the inflection point (ti) are lower than the maximum fat deposition rate in both genotypes. Regarding the fatty acid profile of the carcass, the content of total and individual saturated fatty acids was similar (p > 0.05) in the two genotypes was observed. However, the content of total monounsaturated fatty acids and oleic acid (C18:1C) were higher (p < 0.05) in the Cieneguilla genotype, while the content of total polyunsaturated fatty acids, linoleic and linolenic acids were higher (P < 0.05) in the carcass of guinea pigs of the Peru genotype. In conclusion, the asymptote of moisture content and crude protein in adulthood was higher (p < 0.05) in the Peru genotype. In saturated fatty acid content, there were no statistical differences between both genotypes, but the Cieneguilla genotype contains a higher (p < 0.05) percentage of monounsaturated fatty acids and the Peru genotype has a higher (p < 0.05) percentage of polyunsaturated fatty acids.

Variability of Commercial Saw Palmetto–Based Supplements for the Management of Benign Prostatic Hyperplasia/Lower Urinary Tract Symptoms

ABSTRACT Purpose: Some men with mild-to-moderate benign prostatic hyperplasia/lower urinary tract symptoms use saw palmetto supplements to proactively manage their symptoms as an alternative to watchful waiting and/or to avoid prescription medication side effects. This study assessed the potency and authenticity of commercially available saw palmetto–based supplements in the United States. Materials and Methods: Twenty-eight saw palmetto berry powders, powdered extracts, berry blends, lipid extracts, and multiactive products (lycopene, pumpkin oil, etc) were purchased from major online retailers and retail stores. Total fatty acid content (% weight/weight) and individual fatty acid profile of each product were determined using validated gas chromatography-fatty acid methyl ester methodology and compared with the US Pharmacopeia monograph standards for lipidosterolic extracts of Serenoa repens. Results: Total fatty acid content ranged from 0.796% for a berry powder product to 89.923% for a lipid extract product. None of the berry powders or powdered extracts, 6 of 9 lipid extracts, and 1 multiactive product met criteria for ≥80% total fatty acid content. Only 1 of the 28 products met the US Pharmacopeia criteria for a standardized lipidosterolic extract, defined as total fatty acid content ≥80% and a fatty acid profile indicative of authentic S. repens based on the ratios of the lauric acid concentration to 9 other individual fatty acid concentrations. Conclusions: There is substantial heterogeneity in fatty acid content and profile in saw palmetto supplements. Lipidosterolic extracts of saw palmetto berries standardized to ≥80% fatty acids are most likely to meet established criteria for quality and identity.

Fatty acid profile of traditional Slovak ewe’s and cow’s lump cheese

ABSTRACT The main goal of the research was to find out how the profile and content of individual fatty acids changes in Slovak ewe and cow’s lump cheeses (TSG – Traditional Specialty Guaranteed by European Union). Local farmer produced the cheese. We have analyzed 94 samples of cheeses produced during the production season by the GC/FID method. Fatty acids were found in various concentrations. The following fatty acids were present in the highest concentrations in Ewe’s lump cheese: palmitic acid C16:0 (25.67–35.59 g/100 g), oleic acid C18:1 (n-9) (16.26–25.92 g 100 g−1), myristic acid C14:0 (10.15–14.41 g 100 g−1) and and stearic acid C18:0 (7.23–10.95 g 100 g−1); in Cow’s lump cheese palmitic acid C16:0 (29.46–36.94 g/100 g), oleic acid C18:1 (n-9) (18.93–26.54 g 100 g−1), myristic acid C14:0 (10.07–13.26 g 100 g−1 and stearic acid C18:0 (8.13–10.44 g 100 g−1). Significant (p < .005) changes in individual fatty acid concentrations was found over the production season for most fatty acids. Small farmers can learn the technological process of producing TSG products, but it is important to consider particular seasonal changes in the quantity of fat and fatty acids. From a nutritional perspective, high vaccenic acid concentrations are advantageous, whereas high saturated myristic and palmitic acid concentrations are dubious. The results of this research can be used to supplement the national specification of the TSG Slovak traditional Ewe’s lump cheese (Ovčí hrudkový syr – salašnícky).

Effects of corn silage and grain expressing α-amylase on ruminal nutrient digestibility, microbial protein synthesis, and enteric methane emissions in lactating cows

Increasing ruminal starch digestibility has the potential to improve microbial protein synthesis (MPS), milk production, and feed efficiency. Enogen corn (Syngenta Seeds LLC) expresses high α-amylase activity, and we evaluated effects of Enogen corn silage (CS) and grain (CG) on ruminal starch digestibility, MPS, and milk production in lactating dairy cows. Fifteen Holstein cows (6 ruminally cannulated and 9 noncannulated; average ± standard deviation at the beginning of the trial: 170 ± 40 d in milk; milk yield, 37.2 ± 7.73 kg/d; body weight, 714 ± 37 kg) were used in a replicated 3 × 3 Latin square design (28 d per period) with 3 treatments: a diet containing isoline CS and CG (control, CON); a diet with Enogen CS and isoline CG (ECS); and a diet with Enogen CS and CG (ECSCG). Dry matter (DM; 30%), starch (35% of DM), and particle size distribution of the isoline and Enogen CS were similar. However, the mean particle size of Enogen CG was larger (1.05 vs. 0.65 mm) than that of the isoline CG. Cannulated cows were used for digestibility and nutrient flow measurements, noncannulated cows were used for enteric CH4 measurements, and all cows were used for production evaluation. Dry matter intake (DMI) and milk yield were greater for ECS and ECSCG compared with CON (26.7 and 26.6 vs. 25.1 kg/d and 36.5 and 34.1 vs. 33.1 kg/d, respectively) without a difference between ECS and ECSCG. Milk protein yield was greater (1.27 vs. 1.14 and 1.17 kg/d) for ECS compared with CON and ECSCG. Milk fat content was greater (3.79 vs. 3.32%) for ECSCG compared with ECS. Milk fat yield and energy-corrected milk did not differ among treatments. Ruminal digestibilities of DM, organic matter, starch, and neutral detergent fiber were not different among treatments. However, ruminal digestibility of nonammonia, nonmicrobial N was greater (85 vs. 75%) for ECS compared with ECSCG. Total-tract apparent starch digestibility was lower (97.6 and 97.1 vs. 98.3%) for ECS and ECSCG compared with CON, respectively, and tended to be lower (97.1 vs. 98.3%) for ECSCG compared with ECS. Ruminal outflows of bacterial OM and nonammonia N tended to be greater for ECS than for ECSCG. Efficiency of MPS tended to be greater (34.1 vs. 30.6 g of N/kg of organic matter truly digested) for ECS versus ECSCG. Ruminal pH and total and individual short-chain fatty acid concentrations did not differ among treatments. Concentration of ruminal NH3 for ECS and ECSCG was lower (10.4 and 12.4 vs. 13.4 mmol/L, respectively) compared with CON. Methane per unit of DMI decreased for ECS and ECSCG compared with CON (11.4 and 12.2 vs. 13.5 g/kg of DMI, respectively) without a difference between ECS and ECSCG. In conclusion, ECS and ECSCG did not increase ruminal or total-tract starch digestibility. However, the positive effects of ECS and ECSCG on milk protein yield, milk yield, and CH4 per unit of DMI may show potential benefits of feeding Enogen corn. Effects of ECSCG were not apparent when compared with ECS, partly due to larger particle size of Enogen CG compared with its isoline counterpart.

Impatto degli antiossidanti sulla qualità e la stabilità ossidativa dei grassi animali

The aim of the work was to determine the influence of natural and synthetic antioxidants on the quality and stability of animal fats. On the samples, with and without the addition of natural antioxidants (extract of sage, rosemary and ginger) added in the fats at a concentracion of 0.2 % and synthetic antioxidants (PG-propyl galate, BHA-butylhydroxyanisole and BHT-butylhidroxytoluene) added in the fats at a concentracion 0.01 % the values of the peroxide value and the content of free fatty acids were tested. The oxidative stability of fats was determined by the sustainability test at 98 °C during 7 days. Every 24 hours, the peroxide value and free fatty acids were examinedid for all samples of beef, sheep and goat fat. The results of the research show that the applied antioxidants successfully stabilized all animal fats compared to the control sample. Analysis of the composition of fatty acids of base samples (without the addition of natural antioxidants) shows that palmitic, stearic and oleic fatty acids are the most abundant in animal fats. The highest total SFA value was found in sheep fat (66.85 %), and the lowest in goat fat (56.57 %). In beef fat, the total SFA content was 59.26 %. Stearic acid is the most abundant in sheep fat (37.75 %), while beef and goat fat had lower and approximately equal values (27.93 % and 26.91 %). After staeric fat, palmitic fat is the mostabundant in beef fat with 27.78 %, followed by 25.25 % in sheep fat and 24.91 % in goat fat. Of the monounsaturated acids (MUFA), the most abundant in goat fat was oleic acid (38.16 %), while it was somewhat lower in beef (36.61 %), and the lowest in sheep (27.60 %). Total unsaturated fatty acids (UFA) are most abundant in goat fat (43.51 %), followed by beef (40.82 %), while they are significantly less present in sheep's fat (32.95 %). Among polyunsaturated acids (PUFA), the highest content of linoleic acid was found in sheep fat (4.19 %). In beef fat, linoleic acid was 60 % lower compared to sheep fat (0.76 %), and 10% higher compared to goat fat (1.89 %). Based on the content of individual fatty acids and the content of certain groups of fatty acids: SFA, UFA, MUFA, PUFA, the nutritional indices of the tested fats were calculated. It was concluded that the tested samples of beef, sheep, and goat fat without the addition of antioxidants can not be declared as products of high nutritional value. The nutritional value expressed through the PUFA/SFA ratio for beef fat was 0,01, and sheep fat and goat fat 0.06.

Analysis of Fatty Acid Composition in Sprouted Grains.

A whole-grain diet is associated with the prevention of metabolic syndromes, including obesity, diabetes, and cardiovascular diseases. Sprouting improves the nutritional profile and bioactive properties of grains, which are important for use as raw ingredients in the food industry. The aim of this review was to examine the lipid and fatty acid composition of germinated grains. The methods discussed include germination and analytical procedures for determining fat and fatty acid contents of grains. The effects of sprouting on the fat content and storage stability of grains were also assessed. Lipid levels ranged from 1.43% to 6.66% in the sprouted grains. The individual fatty acid content of grains changed depending on the germination conditions (17-37 °C, 1-9 days). Limited findings showed that sprouting grains at higher temperatures (20-25 °C) and longer times generated a healthy balance of omega-6 and omega-3 fatty acids, which is beneficial to humans. Future studies are needed to determine the optimum incubation and germination periods specific to each grain to improve the omega-6/omega-3 ratio. Free fatty acids were produced more slowly and levels of oxidation products were lower in sprouted grains than in the raw ingredients when stored for a year. Additional studies are required to investigate the oxidative stability and shelf life of sprouted grains.

Effect of replacement of soybean oil by Hermetia illucens fat on performance, digestibility, cecal microbiome, liver transcriptome and liver and plasma lipidomes of broilers

BackgroundIn contrast to protein-rich insect meal, the feed potential of insect fat is generally less explored and knowledge about the suitability of insect fat as a fat source specifically in broiler diets is still limited. In view of this, the present study aimed to comprehensively investigate the effect of partial (50%) and complete replacement of soybean oil with insect fat from Hermetia illucens (HI) larvae in broiler diets on performance, fat digestibility, cecal microbiome, liver transcriptome and liver and plasma lipidomes. Thus, 100 male, 1-day-old Cobb 500 broilers were randomly assigned to three groups and fed three different diets with either 0 (group HI-0, n = 30), 2.5% (group HI-2.5, n = 35) or 5.0% (HI-5.0, n = 35) Hermetia illucens (HI) larvae fat for 35 d.ResultsBody weight gain, final body weight, feed intake, and feed:gain ratio during the whole period and apparent ileal digestibility coefficient for ether extract were not different between groups. Cecal microbial diversity did not differ between groups and taxonomic analysis revealed differences in the abundance of only four low-abundance bacterial taxa among groups; the abundances of phylum Actinobacteriota, class Coriobacteriia, order Coriobacteriales and family Eggerthellaceae were lower in group HI-5.0 compared to group HI-2.5 (P < 0.05). Concentrations of total and individual short-chain fatty acids in the cecal digesta were not different between the three groups. Liver transcriptomics revealed a total of 55 and 25 transcripts to be differentially expressed between groups HI-5.0 vs. HI-0 and groups HI-2.5 vs. HI-0, respectively (P < 0.05). The concentrations of most lipid classes, with the exception of phosphatidylethanolamine, phosphatidylglycerol and lysophosphatidylcholine in the liver and cholesterylester and ceramide in plasma (P < 0.05), and of the sum of all lipid classes were not different between groups.ConclusionsPartial and complete replacement of soybean oil with HI larvae fat in broiler diets had no effect on growth performance and only modest, but no adverse effects on the cecal microbiome and the metabolic health of broilers. This suggests that HI larvae fat can be used as an alternative fat source in broiler diets, thereby, making broiler production more sustainable.

Effect of Feeding Different Percentages of Acacia saligna Foliage Hay on Barki Ewes Performance

The study was conducted to evaluate the effect of replacing berseem hay (BH) with dried Acacia saligna foliage (AFH) on the performance and some blood parameters of pregnant Barki ewes. Thirty-Two aged 2-3 years old at the last third of pregnancy (eight in each group, 40.5 ± 0.96 Kg body weight) were fed 0 (Control), 25, 50 and 75% of AFH in replacement of BH. All animals fed 2% of their body weight concentrate feed mixture and ad libitum quantities of BH or AFH forages for 84 days. Average daily change in body weight of ewesduring lactation, milk yield and composition, rumen liquor parameters (pH, volatile fatty acids, ammonia-N and protozoa count), lamb’s growth performance and some serum biochemical parameters weredetermined.Economic evaluation for the tested levels of forages was also estimated. Data were statistically analyzed using SAS computer program software utilizing the general linear model (GLM) procedure. The findings revealed that while AFH had greater neutral detergent fiber (NDF), acid detergent fiber (ADF) andcrude proteins (CP) values than BH, it had lower amounts of dry matter (DM), crude fiber (CF), ether extracts (EE) and fiber fractions. Total DM intake (g/h/day or g/Kg W ) of lactating ewes did not influence significantly by AFH 0.75 replacement in the diets. The replacement of BH with AFH at the rate of 75% resulted in increasing (P&lt;0.05) individual (Acetic, butyric and propionic) and total volatile fatty acids (VFA) concentrations compared with the control treatment. The tested incorporation levels of AFH did not influence the live body weight during the lactation period. Neglecting the lamb's sex, the tested levels of AFH had noeffect onbirth weight and body weight changes from birth till weaning. Ewes fed 75% AFH recorded the highest total milk yield and daily milk yield compared to the other tested diets.Treatmentsaffects insignificantlyfat and protein yields. Moreover, no significant effect noticed among the experimental groups in serum biochemical tested parameters. Ewes fed ration containing 75% AFH recorded the best economic efficiency compared to other tested rations.

ELOVL5 and IGFBP6 genes modulate sensitivity of breast cancer cells to ferroptosis.

Introduction: Relapse of breast cancer is one of the key obstacles to successful treatment. Previously we have shown that low expression of ELOVL5 and IGFBP6 genes in breast cancer tissue corresponded to poor prognosis. ELOVL5 participates directly in the elongation of polyunsaturated fatty acids (PUFAs) that are considered to play an important role in cancer cell metabolism. Thus, in this work we studied the changes in lipid metabolism in breast cancer cells with reduced expression of either ELOVL5 or IGFBP6 gene. Methods: MDA-MB-231 cells with a stable knockdown of either ELOVL5 or IGFBP6 gene were used in this study. Transcriptomic and proteomic analysis as well as RT-PCR were utilized to assess gene expression. Content of individual fatty acids in the cells was measured with HPLC-MS. HPLC was used for analysis of the kinetics of PUFAs uptake. Cell viability was measured with MTS assay. Flow cytometry was used to measure activation of apoptosis. Fluorescent microscopy was utilized to assess accumulation of ROS and formation of lipid droplets. Glutathione peroxidase activity was measured with a colorimetric assay. Results: We found that the knockdown of IGFBP6 gene led to significant changes in the profile of fatty acids in the cells and in the expression of many genes associated with lipid metabolism. As some PUFAs are known to inhibit proliferation and cause death of cancer cells, we also tested the response of the cells to single PUFAs and to combinations of docosahexaenoic acid (DHA, a n-3 PUFA) with standard chemotherapeutic drugs. Our data suggest that external PUFAs cause cell death by activation of ferroptosis, an iron-dependent mechanism of cell death with excessive lipid peroxidation. Moreover, both knockdowns increased cells' sensitivity to ferroptosis, probably due to a significant decrease in the activity of the antioxidant enzyme GPX4. Addition of DHA to commonly used chemotherapeutic drugs enhanced their effect significantly, especially for the cells with low expression of IGFBP6 gene. Discussion: The results of this study suggest that addition of PUFAs to the treatment regimen for the patients with low expression of IGFBP6 and ELOVL5 genes can be potentially beneficial and is worth testing in a clinically relevant setting.

Network analysis to evaluate complexities in relationships among fermentation variables measured within continuous culture experiments.

The objective of this study was to leverage a frequentist (ELN) and Bayesian learning (BLN) network analyses to summarize quantitative associations among variables measured in 4 previously published dual-flow continuous culture fermentation experiments. Experiments were originally designed to evaluate effects of nitrate, defaunation, yeast, and/or physiological shifts associated with pH or solids passage rates on rumen conditions. Measurements from these experiments that were used as nodes within the networks included concentrations of individual volatile fatty acids, mM and nitrate, NO3-,%; outflows of non-ammonia nitrogen (NAN, g/d), bacterial N (BN, g/d), residual N (RN, g/d), and ammonia N (NH3-N, mg/dL); degradability of neutral detergent fiber (NDFd, %) and degradability of organic matter (OMd, %); dry matter intake (DMI, kg/d); urea in buffer (%); fluid passage rate (FF, L/d); total protozoa count (PZ, cells/mL); and methane production (CH4, mmol/d). A frequentist network (ELN) derived using a graphical LASSO (least absolute shrinkage and selection operator) technique with tuning parameters selected by Extended Bayesian Information Criteria (EBIC) and a BLN were constructed from these data. The illustrated associations in the ELN were unidirectional yet assisted in identifying prominent relationships within the rumen that were largely consistent with current understanding of fermentation mechanisms. Another advantage of the ELN approach was that it focused on understanding the role of individual nodes within the network. Such understanding may be critical in exploring candidates for biomarkers, indicator variables, model targets, or other measurement-focused explorations. As an example, acetate was highly central in the network suggesting it may be a strong candidate as a rumen biomarker. Alternatively, the major advantage of the BLN was its unique ability to imply causal directionality in relationships. Because the BLN identified directional, cascading relationships, this analytics approach was uniquely suited to exploring the edges within the network as a strategy to direct future work researching mechanisms of fermentation. For example, in the BLN acetate responded to treatment conditions such as the source of N used and the quantity of substrate provided, while acetate drove changes in the protozoal populations, non-NH3-N and residual N flows. In conclusion, the analyses exhibit complementary strengths in supporting inference on the connectedness and directionality of quantitative associations among fermentation variables that may be useful in driving future studies.

Solvent-Free Lipid Separation and Attenuated Total Reflectance Infrared Spectroscopy for Fast and Green Fatty Acid Profiling of Human Milk

This study presents the first mid-infrared (IR)-based method capable of simultaneously predicting concentrations of individual fatty acids (FAs) and relevant sum parameters in human milk (HM). Representative fat fractions of 50 HM samples were obtained by rapid, two-step centrifugation and subsequently measured with attenuated total reflection IR spectroscopy. Partial least squares models were compiled for the acquired IR spectra with gas chromatography-mass spectrometry (GC-MS) reference data. External validation showed good results particularly for the most important FA sum parameters and the following individual FAs: C12:0 (R2P = 0.96), C16:0 (R2P = 0.88), C18:1cis (R2P = 0.92), and C18:2cis (R2P = 0.92). Based on the obtained results, the effect of different clinical parameters on the HM FA profile was investigated, indicating a change of certain sum parameters over the course of lactation. Finally, assessment of the method's greenness revealed clear superiority compared to GC-MS methods. The reported method thus represents a high-throughput, green alternative to resource-intensive established techniques.

Seed Weight and Genotype Influence the Total Oil Content and Fatty Acid Composition of Peanut Seeds.

Peanut, an important oilseed crop cultivated worldwide as a dietary food, is a good food source with health benefits. To explore the potential benefits of peanuts as a food resource, 301 peanut accessions were evaluated to determine the effect of seed weight and genotype on total oil content and fatty acid composition. Total oil was extracted using the Soxhlet method and fatty acids were analyzed by gas chromatography mass spectrometry. Wide variations in the 100-seed weight, total oil content, and fatty acid profile were observed among genotypes and accession types. An effect of seed weight on the fatty acid composition of peanut seeds was observed. Increases in the oleic acid content and decreases in the linoleic acid content occurred in association with increases in the 100-seed weight. Moreover, the 100-seed weight, total oil content, and individual and total fatty acid contents, except arachidic acid, differed significantly (p &lt; 0.001 or 0.05) among the accession types of landrace, cultivar, breeding line, and unknown. The discovery of this high diversity could contribute to further studies of peanut domestication and evolutionary classification. Our findings are important for the selection of peanut seeds with health benefits and development of new varieties of peanut with health benefits.

Metabolic fingerprinting of dogs with idiopathic epilepsy receiving a ketogenic medium-chain triglyceride (MCT) oil.

Consumption of medium-chain triglycerides (MCT) has been shown to improve seizure control, reduce behavioural comorbidities and improve cognitive function in epileptic dogs. However, the exact metabolic pathways affected by dietary MCT remain poorly understood. In this study, we aimed to identify changes in the metabolome and neurotransmitters levels relevant to epilepsy and behavioural comorbidities associated with the consuming of an MCT supplement (MCT-DS) in dogs with idiopathic epilepsy (IE). Metabolic alterations induced by a commercial MCT-DS in a population of 28 dogs with IE were evaluated in a 6-month multi-centre, prospective, randomised, double-blinded, controlled cross-over trial design. A metabolic energy requirement-based amount of 9% MCT or control oil was supplemented to the dogs' stable base diet for 3 months, followed by the alternative oil for another 3 months. A validated, quantitative nuclear magnetic resonance (NMR) spectroscopy platform was applied to pre- and postprandially collected serum samples to compare the metabolic profile between both DS and baseline. Furthermore, alterations in urinary neurotransmitter levels were explored. Five dogs (30%) had an overall reduction in seizure frequency of ≥50%, and were classified as MCT-responders, while 23 dogs showed a ≤50% reduction, and were defined as MCT non-responders. Amino-acid metabolism was significantly influenced by MCT consumption compared to the control oil. While the serum concentrations of total fatty acids appeared similar during both supplements, the relative concentrations of individual fatty acids differed. During MCT supplementation, the concentrations of polyunsaturated fatty acids and arachidonic acid were significantly higher than under the control oil. β-Hydroxybutyric acid levels were significantly higher under MCT supplementation. In total, four out of nine neurotransmitters were significantly altered: a significantly increased γ-aminobutyric acid (GABA) concentration was detected during the MCT-phase accompanied by a significant shift of the GABA-glutamate balance. MCT-Responders had significantly lowered urinary concentrations of histamine, glutamate, and serotonin under MCT consumption. In conclusion, these novel data highlight metabolic changes in lipid, amino-acid and ketone metabolism due to MCT supplementation. Understanding the metabolic response to MCT provides new avenues to develop better nutritional management with improved anti-seizure and neuroprotective effects for dogs with epilepsy, and other behavioural disorders.

Effect of Breed on the Fatty Acid Composition of Milk from Dairy Cows Milked Once and Twice a Day in Different Stages of Lactation

The objective of this study was to evaluate the effect of breed on the overall composition and fatty acid composition of milk from cows milked once a day (OAD) and twice a day (TAD) in different stages of lactation. Milk samples were taken from 39 Holstein-Friesian (F), 27 Jersey (J), and 34 Holstein-Friesian × Jersey (F × J) crossbred cows from a OAD milking herd and 104 F and 83 F × J cows from a TAD milking herd in early (49 ± 15 days in milk), mid (129 ± 12 days in milk), and late (229 ± 13 days in milk) lactation. Calibration equations to predict the concentrations of individual fatty acids were developed using mid-infrared (MIR) spectroscopy. There was a significant interaction between breed within the milking frequency and stage of lactation for the production traits and composition traits. Holstein-Friesian cows milked OAD produced milk with lower concentrations of C18:0 in early and mid lactations compared to F × J and J cows. Holstein-Friesian cows milked TAD produced lower concentrations of C18:0 in early lactation and lower concentrations of C16:0 and C18:0 in late lactation compared to F × J. Lower concentrations of these fatty acids would reduce the hardness of the butter when the milk is processed. In the OAD milking herd, F cows were superior for daily milk yield compared to J cows, but Jersey cows produced significantly (p &lt; 0.05) higher percentages of fat and a higher concentration of C18:0 fatty acid. The relative concentrations of C18:0 and C18 cis-9 in F and J cows milked OAD imply there is no breed effect on the activity of delta-9-desaturase, whereas stages of lactation likely have an effect. These results can be used to assist with selecting breeds and cows that are suitable for either OAD or TAD milking, allowing closer alignment with milk processing needs.

In vitro and in vivo Studies of Soybean Peptides on Milk Production, Rumen Fermentation, Ruminal Bacterial Community, and Blood Parameters in Lactating Dairy Cows.

Soybean peptides (SPs), a feed additive derived from soybean, exhibit nutritional function and biological activity in monogastric animals, but limited studies have been conducted in dairy cows. Our experiments were conducted to evaluate the effects of SPs on the nutrient degradability of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF) in vitro and milk production, rumen fermentation and bacterial community, and blood parameters of dairy cows. For in vitro experiment, ruminal fluids were collected from three ruminal cannulated Holstein dairy cows. A total of three levels of SPs (0, 0.38, and 1.92 g/kg DM of SPs) were added to the total mixed ration (TMR). Nutrient degradability and fermentation fluid pH were determined at 24 and 48 h using 3.0 g samples of the substrate. Gas production after 48 h was recorded by an automated trace gas recording system using 0.5 g samples of the substrate. The results showed that DM, NDF, ADF (p < 0.01), and CP (p < 0.05) degradabilities were significantly increased at 1.92 g/kg DM of SPs at 24 h, and asymptotic gas production (p = 0.05) was increased at 48 h. For in vivo experiment, 110 lactating Holstein cows (209.7 ± 65.2 DIM; 37.2 ± 6.4 kg/d milk yield) were randomly assigned to 0 (control group, CON) or 50 g/head/day SPs (SP-supplemented group). Yields of milk (p < 0.05), milk protein (p < 0.05), and milk lactose (0.05 < p < 0.10) increased on SPs supplementation; however, the milk fat percentage decreased (p < 0.05). The concentrations of individual volatile fatty acids (VFAs) (p < 0.05) and superoxide dismutase (SOD) (p < 0.01) were also increased. Rumen bacterial diversity in SP-supplemented cows was higher (p < 0.05). The relative abundances of Rikenellaceae_RC9_gut_group, Butyrivibrio, Selenomonas, and Shuttleworthia were significantly increased and that of Coprococcus was decreased (p < 0.05). Our results showed that supplementing 1.92 g/kg DM of SPs could improve the nutrient degradability in vitro and 50 g/head/day of SPs could improve milk production and antioxidant ability of dairy cows. The rumen bacterial diversity was also enhanced by SP supplementation.