Fatty Acid Pool Research Articles

Temporal variation in the biochemical ecology of lower trophic levels in the Northern California Current

There is strong correlative evidence that variation in the growth and survival of secondary consumers is related to the copepod species composition within the Northern California Current. Potential mechanisms driving these correlations include: (1) enhanced growth and survival of secondary consumers when lipid-rich, boreal copepod species are abundant, with cascading effects on higher trophic levels; (2) the regulation of growth and condition of primary and secondary consumers by the relative proportion of certain essential fatty acids (FAs) in primary producers; or (3) a combination of these factors. Disentangling the relative importance of taxonomic composition, lipid quantity, and FA composition on the nutritional quality of copepods requires detailed information on both the consumer and primary producers. Therefore, we collected phytoplankton and copepods at an oceanographic station for 19months and completed species community analyses and generated detailed lipid profiles, including lipid classes and FAs, for both groups. There was strong covariation between species and biochemistry within and across trophic levels and distinct seasonal differences. The amount of total lipid within both the phytoplankton and copepod communities was twice as high in spring and summer than in fall and winter, and certain FAs, such as diatom indicators 20:5ω3 and 16:1ω7, comprised a greater proportion of the FA pool in spring and summer. Indicators of bacterial production within the copepod community were proportionally twice as high during fall and winter than spring and summer. Seasonal transitions in copepod FA composition were consistently offset from transitions in copepod species composition by approximately two weeks. The timing of the seasonal transition in copepod FAs reflected seasonal shifts in the species composition and/or biochemistry of primary producers more than seasonal shifts in the copepod species composition. These results emphasize the importance of interactions between the copepod community and their available phytoplankton prey in regulating the nutritional quality of primary consumers.

Clinical Relevance of Type II Fatty Acid Synthesis Bypass in Staphylococcus aureus.

The need for new antimicrobials to treat bacterial infections has led to the use of type II fatty acid synthesis (FASII) enzymes as front-line targets. However, recent studies suggest that FASII inhibitors may not work against the opportunist pathogen Staphylococcus aureus, as environmental fatty acids favor emergence of multi-anti-FASII resistance. As fatty acids are abundant in the host and one FASII inhibitor, triclosan, is widespread, we investigated whether fatty acid pools impact resistance in clinical and veterinary S. aureus isolates. Simple addition of fatty acids to the screening medium led to a 50% increase in triclosan resistance, as tested in 700 isolates. Moreover, nonculturable triclosan-resistant fatty acid auxotrophs, which escape detection under routine conditions, were uncovered in primary patient samples. FASII bypass in selected isolates correlated with polymorphisms in the acc and fabD loci. We conclude that fatty-acid-dependent strategies to escape FASII inhibition are common among S. aureus isolates and correlate with anti-FASII resistance and emergence of nonculturable variants.

Depletion of TM6SF2 disturbs membrane lipid composition and dynamics in HuH7 hepatoma cells

A polymorphism of TM6SF2 associates with hepatic lipid accumulation and reduction of triacylglycerol (TAG) secretion, but the function of the encoded protein has remained enigmatic. We studied the effect of stable TM6SF2 knock-down on the lipid content and composition, mitochondrial fatty acid oxidation and organelle structure of HuH7 hepatoma cells. Knock-down of TM6SF2 resulted in intracellular accumulation of TAGs, cholesterol esters, phosphatidylcholine (PC) and phosphatidylethanolamine. In all of these lipid classes, polyunsaturated lipid species were significantly reduced while saturated and monounsaturated species increased their proportions. The PCs encountered relative and absolute arachidonic acid (AA, 20:4n-6) depletion, and AA was also reduced in the total cellular fatty acid pool. Synthesis and turnover of the hepatocellular glycerolipids was enhanced. The TM6SF2 knock-down cells secreted lipoprotein-like particles with a smaller diameter than in the controls, and more lysosome/endosome structures appeared in the knock-down cells. The mitochondrial capacity for palmitate oxidation was significantly reduced. These observations provide novel clues to TM6SF2 function and raise altered mebrane lipid composition and dynamics among the mechanism(s) by which the protein deficiency disturbs hepatic TAG secretion.

Dynamic regulation of fatty acid pools for improved production of fatty alcohols in Saccharomyces cerevisiae

BackgroundIn vivo production of fatty acid-derived chemicals in Saccharomyces cerevisiae requires strategies to increase the intracellular supply of either acyl-CoA or free fatty acids (FFAs), since their cytosolic concentrations are quite low in a natural state for this organism. Deletion of the fatty acyl-CoA synthetase genes FAA1 and FAA4 is an effective and straightforward way to disable re-activation of fatty acids and drastically increase FFA levels. However, this strategy causes FFA over-accumulation and consequential release to the extracellular medium, which results in a significant loss of precursors that compromises the process yield. In the present study, we aimed for dynamic expression of the fatty acyl-CoA synthetase gene FAA1 to regulate FFA and acyl-CoA pools in order to improve fatty alcohol production yields.ResultsWe analyzed the metabolite dynamics of a faa1Δ faa4Δ strain constitutively expressing a carboxylic acid reductase from Mycobacterium marinum (MmCAR) and an endogenous alcohol dehydrogenase (Adh5) for in vivo production of fatty alcohols from FFAs. We observed production of fatty acids and fatty alcohols with different rates leading to high levels of FFAs not being converted to the final product. To address the issue, we expressed the MmCAR + Adh5 pathway together with a fatty acyl-CoA reductase from Marinobacter aquaeolei to enable fatty alcohol production simultaneously from FFA and acyl-CoA, respectively. Then, we expressed FAA1 under the control of different promoters in order to balance FFA and acyl-CoA interconversion rates and to achieve optimal levels for conversion to fatty alcohols. Expressing FAA1 under control of the HXT1 promoter led to an increased accumulation of fatty alcohols per OD600 up to 41% while FFA levels were decreased by 63% compared with the control strain.ConclusionsFine-tuning and dynamic regulation of key metabolic steps can be used to improve cell factories when the rates of downstream reactions are limiting. This avoids loss of precursors to the extracellular medium or to competing reactions, hereby potentially improving the process yield. The study also provides knowledge of a key point of fatty acid regulation and homeostasis, which can be used for future design of cells factories for fatty acid-derived chemicals.

The parallel comparative study of metabolism of fatty acids in erythrocytes and other tissues under experimental diabetes mellitus

The parallel study of impact of Streptozotocin diabetes mellitus on level of lipids and content of fatty acids in blood, in homogenates of skin, liver and muscular tissue in conditions of oxidative stress demonstrated that in rats at early period of development of diabetes mellitus alongside with hypertriglyceridimia and hypercholesterolimia, significant alterations are observed in content of fatty acids of blood and homogenates of analyzed tissues. The alterations are accompanied with peroxidation of lipids and decreasing of activity of enzymes-antioxidants and also decreasing of degree of utilization of glucose by erythrocytes. under experimental diabetes mellitus significant increasing of saturated fatty acids, especially palmitic fatty acid, results in decreasing of level of mono unsaturated fatty acids (at the expense of oleic fatty acids) that significantly differs of corresponding spectrum in patients with diabetes mellitus. The results of study also testify that already at early stages of development of diabetes mellitus alterations in pool of fatty acids of erythrocytes and homogenates of liver, skin and cardiac muscle similar (with slightly differences) and in all analyzed tissues disorders of fatty acids metabolism are characterized by atherosclerotic direction.

Metabolomic analysis of Salvia hispanica seeds using NMR spectroscopy and multivariate data analysis

The metabolic profile of the seeds of seven Chia (Salvia hispanica L.) populations, three commercial (two black, and one white) and four early flowering genotypes (G3, G8, G17, W13.1), was investigated by NMR spectroscopy. The research aimed at evaluating the chemical composition of the different genotypes both from a qualitative point of view, through the identification of the major classes of organic compounds by NMR analysis, and from a quantitative point of view, through the integration of NMR spectra followed by Principal Component Analysis and chemometrics. Results showed that apolar organic extracts were mainly composed of mono- and polyunsaturated fatty acids, such as α-linolenic acid, while polar organic extracts contained the sugars glucose, raffinose and sucrose as the main metabolites, together with caffeoyl derivatives, flavonoids, organic acids, and free amino acids. Tashinone I and 15,16 dihydro Tanshinone I were detected in Chia seeds for the first time. Chemometric results showed significant differences in the metabolic fingerprinting of the different populations, with the content of most of the detected metabolites showing higher variation in the seeds of early flowering genotypes compared to commercial seeds. In particular, black genotypes are richest in carbohydrates comparing to the early flowering and white genotypes. The analysis was also extended to two black samples grown in Basilicata (Southern Italy) to evaluate the effect of agronomic management, such as fertilization with mineral nitrogen, on the metabolite composition. The obtained data indicated that the effect of mineral nitrogen supply positively affected the content of aliphatic free amino acids, and negatively that of the main carbohydrates and flavonoids, while the pools of caffeoyl derivatives, organic and fatty acids remained almost unaffected.

Effect of initial pH of medium with potato wastewater and glycerol on protein, lipid and carotenoid biosynthesis by Rhodotorula glutinis

Background: Rhodotorula glutinis yeast is capable of the synthesis of numerous valuable metabolites with extensive potential industrial usage. This paper reports the study on determination of the effect of initial culture medium pH on growth and protein, lipid and carotenoid biosynthesis by R. glutinis yeast. Results: The highest biomass yield was obtained in media at pH 4.0-7.0, and the value after 72 h was 17.2-19.4 g d.w. /L. An initial pH of the medium in the range 4.0-7.0 have no significant effect on the protein (38.5-41.3 g/100 g d.w. ), lipid (10.2-12.7 g/100 g d.w. ), or carotenoid (191.7-202.9 μg/g d.w. ) content in the biomass, or on the profile of synthesized fatty acids and carotenoids. The whole pool of fatty acids was dominated by oleic (48.1-53.4%), linoleic (21.4-25.1%) and palmitic acids (13.0-15.8%). In these conditions, the yeast mainly synthesized torulene (43.5-47.7%) and β-carotene (34.7-38.6%), whereas the torularhodin contribution was only 12.1-16.8%. Cultivation in medium at initial pH of 3.0 resulted in a growth reduction (13.0 g d.w. /L) and total carotenoid (115.8 µg/g d.w. ), linoleic acid (11.5%) and torularhodin (4.5%) biosynthesis. Conclusion: The different values of initial pH of the culture medium with glycerol and deproteinized potato wastewater had a significant effect on the growth and on protein, lipid and carotenoid biosynthesis by R. glutinis yeast. Normal 0 false false false EN-GB X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:Tabla normal; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:8.0pt; mso-para-margin-left:0cm; line-height:107%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:Calibri,sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Times New Roman; mso-bidi-theme-font:minor-bidi;}

Effects of prolonged darkness and temperature on the lipid metabolism in the benthic diatom Navicula perminuta from the Arctic Adventfjorden, Svalbard

The Arctic represents an extreme habitat for phototrophic algae due to long periods of darkness caused by the polar night (~4 months darkness). Benthic diatoms, which dominate microphytobenthic communities in shallow water regions, can survive this dark period, but the underlying physiological and biochemical mechanisms are not well understood. One of the potential mechanisms for long-term dark survival is the utilisation of stored energy products in combination with a reduced basic metabolism. In recent years, water temperatures in the Arctic increased due to an ongoing global warming. Higher temperatures could enhance the cellular energy requirements for the maintenance metabolism during darkness and, therefore, accelerate the consumption of lipid reserves. In this study, we investigated the macromolecular ratios and the lipid content and composition of Navicula cf. perminuta Grunow, an Arctic benthic diatom isolated from the microphytobenthos of Adventfjorden (Svalbard, Norway), over a dark period of 8 weeks at two different temperatures (0 and 7 °C). The results demonstrate that N. perminuta uses the stored lipid compound triacylglycerol (TAG) during prolonged dark periods, but also the pool of free fatty acids (FFA). Under the enhanced temperature of 7 °C, the lipid resources were used significantly faster than at 0 °C, which could consequently lead to a depletion of this energy reserves before the end of the polar night. On the other hand, the membrane building phospho- and glycolipids remained unchanged during the 8 weeks darkness, indicating still intact thylakoid membranes. These results explain the shorter survival times of polar diatoms with increasing water temperatures during prolonged dark periods.

Turnover of microbial groups and cell components in soil: <sup>13</sup>C analysis of cellular biomarkers

Abstract. Microorganisms regulate the carbon (C) cycle in soil, controlling the utilization and recycling of organic substances. To reveal the contribution of particular microbial groups to C utilization and turnover within the microbial cells, the fate of 13C-labelled glucose was studied under field conditions. Glucose-derived 13C was traced in cytosol, amino sugars and phospholipid fatty acid (PLFA) pools at intervals of 3, 10 and 50 days after glucose addition into the soil. 13C enrichment in PLFAs ( ∼ 1.5 % of PLFA C at day 3) was an order of magnitude greater than in cytosol, showing the importance of cell membranes for initial C utilization. The 13C enrichment in amino sugars of living microorganisms at day 3 accounted for 0.57 % of total C pool; as a result, we infer that the replacement of C in cell wall components is 3 times slower than that of cell membranes. The C turnover time in the cytosol (150 days) was 3 times longer than in PLFAs (47 days). Consequently, even though the cytosol pool has the fastest processing rates compared to other cellular compartments, intensive recycling of components here leads to a long C turnover time. Both PLFA and amino-sugar profiles indicated that bacteria dominated in glucose utilization. 13C enrichment decreased with time for bacterial cell membrane components, but it remained constant or even increased for filamentous microorganisms. 13C enrichment of muramic acid was the 3.5 times greater than for galactosamine, showing a more rapid turnover of bacterial cell wall components compared to fungal. Thus, bacteria utilize a greater proportion of low-molecular-weight organic substances, whereas filamentous microorganisms are responsible for further C transformations. Thus, tracing 13C in cellular compounds with contrasting turnover rates elucidated the role of microbial groups and their cellular compartments in C utilization and recycling in soil. The results also reflect that microbial C turnover is not restricted to the death or growth of new cells. Indeed, even within living cells, highly polymeric cell compounds are constantly replaced and renewed. This is especially important for assessing C fluxes in soil and the contribution of C from microbial residues to soil organic matter.

Fatty acid profile, oxidative stability and sensory quality of breast meat from turkeys fed diets with graded levels of flaxseed oil for different periods of time

The effect of partial or complete substitution (2.5% and 5%, respectively) of flaxseed oil for soybean oil on the fatty acid profile, oxidative stability and sensory quality of turkey breast meat was studied. Turkeys were fed experimental diets for 3, 4, 5 or 6 weeks before being slaughtered at 16 weeks of age. The percentages of α-linolenic acid and eicosapentaenoic acid in the total fatty acid pool of meat were determined by dietary intake and feeding duration. Even a prolonged dietary treatment with 5% flaxseed oil did not significantly increase the thiobarbituric acid reactive substances content of the turkey meat and did not have a detrimental effect on the sensory properties. The recommended n-6 : n-3 polyunsaturated fatty acids ratio (<4 : 1) in turkey meat can be easily achieved by feeding turkeys a diet containing 2.5% flaxseed oil for 3 weeks. To maintain the above ratio and increase eicosapentaenoic acid concentrations in the meat, the diet should contain 5% flaxseed oil.

The Fatty Acid Profile and Oxidative Stability of Meat from Turkeys Fed Diets Enriched with n-3 Polyunsaturated Fatty Acids and Dried Fruit Pomaces as a Source of Polyphenols.

The aim of this study was to determine the efficacy of different dietary fruit pomaces in reducing lipid oxidation in the meat of turkeys fed diets with a high content of n-3 polyunsaturated fatty acids (PUFAs). Over a period of 4 weeks before slaughter, turkeys were fed diets with the addition of 5% dried apple, blackcurrant, strawberry and seedless strawberry pomaces (groups AP, BP, SP and SSP, respectively) and 2.5% linseed oil. Pomaces differed in the content (from 5.5 in AP to 43.1 mg/g in SSP) and composition of polyphenols Proanthocyanidins were the main polyphenolic fraction in all pomaces, AP contained flavone glycosides and dihydrochalcones, BP contained anthocyanins, and SP and SSP—ellagitannins. The n-6/n-3 PUFA ratio in all diets was comparable and lower than 2:1. In comparison with groups C and AP, the percentage of n-3 PUFAs in the total fatty acid pool of white meat from the breast muscles of turkeys in groups BP, SP and SSP was significantly higher, proportionally to the higher content of α-linolenic acid in berry pomaces. The fatty acid profile of dark meat from thigh muscles, including the n-6/n-3 PUFA ratio, was similar and lower than 3:1 in all groups. Vitamin A levels in raw breast muscles were higher in group AP than in groups C and BP (P<0.05). The addition of fruit pomaces to turkey diets lowered vitamin E concentrations (P = 0.001) in raw breast muscles relative to group C. Diets supplemented with fruit pomaces significantly lowered the concentration of thiobarbituric acid reactive substances (TBARS) in raw, frozen and cooked meat. Our results indicate that the dietary application of dried fruit pomaces increases the oxidative stability of meat from turkeys fed linseed oil, and strawberry pomace exerted the most desirable effects due to its highest polyphenol content and antioxidant potential.

Fatty Acid Biosynthesis Pathways in Methylomicrobium buryatense 5G(B1)

Methane utilization by methanotrophic bacteria is an attractive application for biotechnological conversion of natural or biogas into high-added-value products. Haloalcaliphilic methanotrophic bacteria belonging to the genus Methylomicrobium are among the most promising strains for methane-based biotechnology, providing easy and inexpensive cultivation, rapid growth, and the availability of established genetic tools. A number of methane bioconversions using these microbial cultures have been discussed, including the derivation of biodiesel, alkanes, and OMEGA-3 supplements. These compounds are derived from bacterial fatty acid pools. Here, we investigate fatty acid biosynthesis in Methylomicrobium buryatense 5G(B1). Most of the genes homologous to typical Type II fatty acid biosynthesis pathways could be annotated by bioinformatics analyses, with the exception of fatty acid transport and regulatory elements. Different approaches for improving fatty acid accumulation were investigated. These studies indicated that both fatty acid degradation and acetyl- and malonyl-CoA levels are bottlenecks for higher level fatty acid production. The best strain generated in this study synthesizes 111 ± 2 mg/gDCW of extractable fatty acids, which is ~20% more than the original strain. A candidate gene for fatty acid biosynthesis regulation, farE, was identified and studied. Its deletion resulted in drastic changes to the fatty acid profile, leading to an increased pool of C18-fatty acid methyl ester. The FarE-regulon was further investigated by RNA-seq analysis of gene expression in farE-knockout mutants and farE-overexpressing strains. These gene profiles highlighted a novel set of enzymes and regulators involved in fatty acid biosynthesis. The gene expression and fatty acid profiles of the different farE-strains support the hypothesis that metabolic fluxes upstream of fatty acid biosynthesis restrict fatty acid production in the methanotroph.

Selected fatty acids as biomarkers of exposure to microdoses of molluscicides in snails Helix pomatia (Gastropoda Pulmonata)

Two stages of selection from a pool of 56 fatty acids analyzed in Helix pomatia yielded a set of 12 biomarker acids undergoing significant changes in contact with three microdoses of toxic substances, i.e. three molluscicides containing metaldehyde, methiocarb, and potassium chloride (PC). The proposed palette of acids, including saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA), determined separately in the foot tissues and hepatopancreas of Gastropoda, can be used in ecotoxicological research as a reliable test of the effect of trace doses of stressors. The final set of the biomarker FA comprised C16:0; C18:0; C23:0; C18:1 n-9; C20:1 n-9; C18:2 n-6; C18:3 n-3; C20:2; C20:4 n-6; C20:5 n-3; C22:4 n-6; and C22:5 n-3. A clear physiological response manifested as changes in the content of fatty acids (FA) was observed in the snails even in the case of the lowest doses of the pollutants. All experimental factors analyzed, i.e., the dose (5, 10, or 15 μl 0.01% w/v concentration) and the type of preparation (metaldehyde, methiocarb or PC), had a significant (p ≤ 0.01) impact on the FA composition of the foot and hepatopancreas. Limitation of the analysis to a narrow pool of reactive FA meets the requirements of parameters of biomarkers of exposure and facilitates and accelerates visualization of the bioindicator organism's response to the presence of the stressor in the environment.

Dynamics of Changes in the Fatty Acid Composition of the Auditory Cortex of the Brain in Rats after Single Audiogenic Convulsions

A small but significant increase in the quantity of lipid peroxidation-resistant saturated and monounsaturated fatty acids was found in the auditory cortex of Krushinskii–Molodkina (KM) rats, which have an inherited predisposition to audiogenic convulsions, as compared with Wistar rats. Assessment of the fatty acid composition of the auditory cortex in KM rats at 1 h and 1, 3, and 14 days after single audiogenic convulsions revealed changes at three days, evidencing significant depletion of the fatty acid pool at this time. At 14 days, the auditory cortex of KM rats showed recovery of most fatty acids. These results may be of value in developing new approaches to eliminating brain damage after convulsive seizures.

CHANGES IN BLOOD FREE FATTY ACID CONTENT AMONG CHILDREN WITH LATENT FORM OF CHRONIC RENAL FAILURE

Aim. To assess the status of blood free fatty acid balance in children with megaureter diagnosed latent form of chronic renal failure. Materials and methods. The study design: prospective, controlled, nonrandomized, one-centered. The study included 40 children, divided into 2 groups. The main group ( n = 22) - patients after surgical intervention for megaureter with developing renal dysfunction. The comparison group - 18 relatively healthy children with a mean age of 6,1 ± 1,3 years. Results. It was revealed by the authors that in patients of the main group statistically significantly raised the content of a number of fatty acids. When conducting correlation analysis between the levels of blood total protein, erythrocytes and leukocytes with free fatty acid pool, it was detected that there are reliable, differently directed strong correlations between some of them. Conclusions. Children, who underwent surgical intervention for megaureter followed by development of latent chronic renal failure, differ from healthy children by both total protein content and qualitative composition of blood free fatty acids.

Novel Molecular Interactions of Acylcarnitines and Fatty Acids with Myoglobin

Previous research has indicated that long-chain fatty acids can bind myoglobin (Mb) in an oxygen-dependent manner. This suggests that oxy-Mb may play an important role in fuel delivery in Mb-rich muscle fibers (e.g. type I fibers and cardiomyocytes), and raises the possibility that Mb also serves as an acylcarnitine-binding protein. We report for the first time the putative interaction and affinity characteristics for different chain lengths of both fatty acids and acylcarnitines with oxy-Mb using molecular dynamic simulations and isothermal titration calorimetry experiments. We found that short- to medium-chain fatty acids or acylcarnitines (ranging from C2:0 to C10:0) fail to achieve a stable conformation with oxy-Mb. Furthermore, our results indicate that C12:0 is the minimum chain length essential for stable binding of either fatty acids or acylcarnitines with oxy-Mb. Importantly, the empirical lipid binding studies were consistent with structural modeling. These results reveal that: (i) the lipid binding affinity for oxy-Mb increases as the chain length increases (i.e. C12:0 to C18:1), (ii) the binding affinities of acylcarnitines are higher when compared with their respective fatty acid counterparts, and (iii) both fatty acids and acylcarnitines bind to oxy-Mb in 1:1 stoichiometry. Taken together, our results support a model in which oxy-Mb is a novel regulator of long-chain acylcarnitine and fatty acid pools in Mb-rich tissues. This has important implications for physiological fuel management during exercise, and relevance to pathophysiological conditions (e.g. fatty acid oxidation disorders and cardiac ischemia) where long-chain acylcarnitine accumulation is evident.

Seasonal changes in the D / H ratio of fatty acids of pelagic microorganisms in the coastal North Sea

Abstract. Culture studies of microorganisms have shown that the hydrogen isotopic composition of fatty acids depends on their metabolism, but there are only few environmental studies available to confirm this observation. Here we studied the seasonal variability of the deuterium-to-hydrogen (D / H) ratio of fatty acids in the coastal Dutch North Sea and compared this with the diversity of the phyto- and bacterioplankton. Over the year, the stable hydrogen isotopic fractionation factor ε between fatty acids and water (εlipid/water) ranged between −172 and −237 ‰, the algal-derived polyunsaturated fatty acid nC20:5 generally being the most D-depleted (−177 to −235 ‰) and nC18:0 the least D-depleted fatty acid (−172 to −210 ‰). The in general highly D-depleted nC20:5 is in agreement with culture studies, which indicates that photoautotrophic microorganisms produce fatty acids which are significantly depleted in D relative to water. The εlipid/water of all fatty acids showed a transient shift towards increased fractionation during the spring phytoplankton bloom, indicated by increasing chlorophyll a concentrations and relative abundance of the nC20:5 polyunsaturated fatty acids, suggesting increased contributions of photoautotrophy. Time periods with decreased fractionation (less negative εlipid/water values) can potentially be explained by an increased contribution of heterotrophy to the fatty acid pool. Our results show that the hydrogen isotopic composition of fatty acids is a promising tool to assess the community metabolism of coastal plankton potentially in combination with the isotopic analysis of more specific biomarker lipids.

Annual cycle and spatial trends in fatty acid composition of suspended particulate organic matter across the Beaufort Sea shelf

Fatty acid profiles of suspended particulate organic matter (POM) were determined over an annual cycle (September 2003 to August 2004) on the Beaufort Sea shelf, Canadian Arctic. Special emphasis was placed on the nutritional quality of the fatty acid pool available to zooplankton by examining spatial and temporal patterns in the proportions of total polyunsaturated fatty acids (PUFA) and the essential fatty acids 22:6n−3 (DHA) and 20:5n−3 (EPA). EPA and DHA were the two most abundant PUFA throughout the study period. A log-ratio multivariate (LRA) analysis revealed strong structure in fatty acid profiles related to season and depth. Dominant fatty acids accounting for the observed trend included 18:5n−3, 18:4n−3, 16:1n−7, 20:5n-3, 18:0 and 20:3n−3. We observed a shift in fatty acid profiles from summer to autumn (e.g., from 16:1n−7 and EPA to 18:5n−3 and 18:4n−3) that likely corresponded to a shift in the relative importance of diatoms versus dinoflagellates, prymnesiophytes and/or prasinophytes to the POM pool. Fatty acid composition during winter was dominated by more refractory saturated fatty acids. A surprising finding was the depth and seasonal trend of 20:3n-3, which was higher in winter, aligned with 18:0 in the LRA, but behaved differently than other n−3 PUFA. We interpret fatty acid profiles during summer to be predominantly driven by phytoplankton inputs, whereas fatty acid profiles in winter were dominated by fatty acids that were left over after consumption and/or were generated by heterotrophs. The highest diatom inputs (EPA, the diatom fatty acid marker), n−3/n−6 ratios, and C16 PUFA index were located in an upwelling region off Cape Bathurst. This study is the first annual time series of fatty acid profiles of POM in Arctic seas, expanding our knowledge of the composition of POM throughout the dark season.

Abstract 1052: Stearoyl-CoA-Desaturase (SCD1) regulates lung cancer stemness via stabilization and nuclear localization of YAP/TAZ

Abstract One of the most common traits of cancer is the radical change of cellular metabolism. A distinctive aspect of this altered metabolic status is the presence of a larger pool of monounsaturated fatty acids (MUFA), the precursors of the components of cell membranes necessary to sustained the rapidly dividing cancer cells. MUFA are largely generated from saturated fatty acids by the action of the two Stearoyl-CoA desaturases isoforms, namely SCD1 and SCD5. Recent evidences suggest that the major isoform SCD1 plays a role in several cancers. Our group has previously reported, using primary adenocarcinoma cell lines from malignant pleural effusions, that SCD1 inhibition selectively kills ALDH positive cells, a marker of cancer stem cells, causes spheroid 3D cultures collapse in vitro and impairs their growth in vivo. These results suggest that SCD1 may be a critical target in lung cancer tumor-initiating cells and further studies are needed to assess the role of this enzyme in lung cancer. An increasing number of literature points to the Hippo (and their effectors YAP/ TAZ) and to the Wnt/β-catenin signaling pathways as key factors in the development of cancer. Notably, SCD1 has been shown to be involved in the secretion and maturation of active wnt ligands, contributing to the activation of the β-catenin signaling. We report here, using primary cell cultures from adenocarcinoma of the lung, that YAP and TAZ are required for the generation of 3D cultures, as silencing of both genes completely abrogated the capability of generating lung cancer spheroids. Moreover, simultaneously silencing of YAP and TAZ reduced the mRNA levels of OCT4, Nanog and ALDH. We demonstrated that both silencing of SCD1 and its pharmacological inhibition determined a decrease in the expression and nuclear localization of both YAP and TAZ. Moreover, when SCD1 was inhibited, YAP and TAZ transcriptional activity was reduced. We also demonstrated that SCD1 blockade induced a dramatic reduction of the Axin2 mRNA level, one of the β-catenin target genes and a decreased in the β-catenin transcriptional activity, without affecting β-catenin protein expression. YAP and TAZ downregulation induced by SCD1 blockade could be rescued by the addition of exogenous wnt3a ligand, thus indicating that the active β-catenin signaling is necessary for YAP and TAZ stabilization. Ultimately, we observed that the ubiquitin ligase β-TrcP is involved in the SCD1 mediated degradation of YAP and TAZ. These data, overall, demonstrate for the first time the involvement of SCD1 in the regulation of the Hippo pathway and β-catenin pathway in lung cancer 3D spheroid cultures, pointing to lipid metabolism as regulator of cancer stem features. Moreover, these results can be the starting point for further studies focused on the regulation of SCD1 in cancer stem cells, suggesting a new perspective for improving chemotherapeutic responses in cancer treatment, centered on SCD1 inhibition. Citation Format: Alessia Noto, Maria Elena Pisanu, Claudia De Vitis, Giovanni Sorrentino, Giannino Del Sal, Alfredo Budillon, Gennaro Ciliberto, Rita Mancini. Stearoyl-CoA-Desaturase (SCD1) regulates lung cancer stemness via stabilization and nuclear localization of YAP/TAZ. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1052.

Influence of pomegranate seed oil and bitter melon aqueous extract on polyunsaturated fatty acids and their lipoxygenase metabolites concentration in serum of rats

Competition with polyunsaturated fatty acids (PUFA) and an impact on eicosanoid biosynthesis may be one of mechanisms of conjugated linolenic acids (CLnA) action. The aim of this study was to investigate the influence of diet supplementation with pomegranate seed oil, containing punicic acid (PA)—one of CLnA isomers, and an aqueous extract of dried bitter melon fruits, administered separately or together, on PUFA and their lipoxygenase metabolites’ concentration in serum of rats. Percentage share of fatty acids was diversified in relation to applied supplementation. PA was only detected in serum of pomegranate seed oil supplemented group, where it was about 1%. Cis-9, trans-11 conjugated linoleic acid (rumenic acid, RA) level tended to increase in group supplemented simultaneously with both dietary supplements whereas its highest share in total fatty acids pool was detected in group receiving solely bitter melon dried fruits aqueous extract. This indicates that consumption of bitter melon tea significantly increased RA content in fatty acids pool in serum. However, pomegranate seed oil elevated procarcinogenic 12-hydroxyeicosatetraenoic acid concentration. Taking into account that pomegranate seed oil and bitter melon dried fruits are dietary supplements accessible worldwide and willingly consumed, the biological significance of this phenomenon should be further investigated. We presume, that there may be a need for some precautions concerning the simultaneous use of these products.