Investigation of Deep-Sea Ecosystems Using Marker Fatty Acids: Sources of Essential Polyunsaturated Fatty Acids in Abyssal Megafauna

Abyssal seafloor ecosystems cover more than 50% of the Earth’s surface. Being formed by mainly heterotrophic organisms, they depend on the flux of particulate organic matter (POM) photosynthetically produced in the surface layer of the ocean. As dead phytoplankton sinks from the euphotic to the abyssal zone, the trophic value of POM and the concentration of essential polyunsaturated fatty acids (PUFA) decrease. This results in pronounced food periodicity and limitations for bottom dwellers. Deep-sea invertebrate seston eaters and surface deposit feeders consume the sinking POM. Other invertebrates utilize different food items that have undergone a trophic upgrade, with PUFA synthesized from saturated and monounsaturated FA. Foraminifera and nematodes can synthesize arachidonic acid (AA), eicosapentaenoic acid (EPA), while some barophylic bacteria produce EPA and/or docosahexaenoic acid. FA analysis of deep-sea invertebrates has shown high levels of PUFA including, in particular, arachidonic acid, bacterial FA, and a vast number of new and uncommon fatty acids such as 21:4(n-7), 22:4(n-8), 23:4(n-9), and 22:5(n-5) characteristic of foraminifera. We suppose that bacteria growing on detritus having a low trophic value provide the first trophic upgrading of organic matter for foraminifera and nematodes. In turn, these metazoans perform the second-stage upgrading for megafauna invertebrates. Deep-sea megafauna, including major members of Echinodermata, Mollusca, and Polychaeta display FA markers characteristic of bacteria, foraminifera, and nematodes and reveal new markers in the food chain.

Fasting hepatic de novo lipogenesis is not reliably assessed using circulating fatty acid markers

ABSTRACT Background Observational studies often infer hepatic de novo lipogenesis (DNL) by measuring circulating fatty acid (FA) markers; however, it remains to be elucidated whether these markers accurately reflect hepatic DNL. Objectives We investigated associations between fasting hepatic DNL and proposed FA markers of DNL in subjects consuming their habitual diet. Methods Fasting hepatic DNL was assessed using 2H2O (deuterated water) in 149 nondiabetic men and women and measuring the synthesis of very low-density lipoprotein triglyceride (VLDL-TG) palmitate. FA markers of blood lipid fractions were determined by gas chromatography. Results Neither the lipogenic index (16:0/18:2n–6) nor the SCD index (16:1n–7/16:0) in VLDL-TG was associated with isotopically assessed DNL (r = 0.13, P = 0.1 and r = −0.08, P = 0.35, respectively). The relative abundances (mol%) of 14:0, 16:0, and 18:0 in VLDL-TG were weakly (r ≤ 0.35) associated with DNL, whereas the abundances of 16:1n–7, 18:1n–7, and 18:1n–9 were not associated. When the cohort was split by median DNL, only the abundances of 14:0 and 18:0 in VLDL-TG could discriminate between subjects having high (11.5%) and low (3.8%) fasting hepatic DNL. Based on a subgroup, FA markers in total plasma TG, plasma cholesteryl esters, plasma phospholipids, and red blood cell phospholipids were generally not associated with DNL. Conclusions The usefulness of circulating FAs as markers of hepatic DNL in healthy individuals consuming their habitual diet is limited due to their inability to discriminate clearly between individuals with low and high fasting hepatic DNL.

Fatty acid composition as an indicator of possible sources of nutrition for soft corals of the genusSinularia(Alcyoniidae)

Fatty acids (FAs) composition of eight zooxanthellate soft corals,Sinularia leptoclados, S. flexibilis, S.aff.deformis, S. lochmodes, S. cf.muralis, S. densa, S. notandaandS. cruciatacollected in Van Phong Bay (Vietnam) were studied to identify possible origin of unsaturated FAs. The main FAs were 14:0, 16:0, 7-Me-16:1n-10, 16:1n-7, 16:2n-7, 18:0, 18:1n-9, 18:4n-3, 20:4n-6, 20:5n-3, 22:6n-3, 24:5n-6 and 24:6n-3. On the average, saturated, monounsaturated, and polyunsaturated FAs (PUFAs) contributed 35.6, 6.2 and 54.0% of total coral FAs, respectively. PUFAs of n-6 series predominated in all animals (n-6/n-3 > 1.6). The content of 20:4n-6 varied from 10.2 to 23.8%. The main n-3 PUFA was 18:4n-3 (on the average, 5.4%); the contribution of 20:5n-3 and 22:6n-3, typical PUFAs of marine organisms, was not more than 2.4 and 3.9%, respectively. InSinularia, PUFAs were produced by endosymbiotic dinoflagellates (zooxanthellae) and the coral host tissue, or obtained with food. Zooxanthellae can be considered as the source of C16PUFAs and 18:4n-3. The coral host synthesized 18:2n-7, 24:5n-6 and 24:6n-3 acids. The low content of 18:1n-7, saturated odd-chain FAs and saturated methyl-branched FAs indicated a negligible contribution of bacteria to total lipids ofSinularia. A comparison of the levels of diatom and dinoflagellate FA markers in coral and plankton lipids showed eukaryotic microalgae to play a secondary role in feeding ofSinularia. The high level of 20:4n-6 may be considered as an indicator of heterotrophic feeding ofSinularia.

Protein kinase G signaling disrupts Rac1-dependent focal adhesion assembly in liver specific pericytes

Nitric oxide (NO) regulates the function of perivascular cells (pericytes), including hepatic stellate cells (HSC), mainly by activating cGMP and cGMP-dependent kinase (PKG) via NO/cGMP paracrine signaling. Although PKG is implicated in integrin-mediated cell adhesion to extracellular matrix, whether or how PKG signaling regulates the assembly of focal adhesion complexes (FA) and migration of HSC is not known. With the help of complementary molecular and cell biological approaches, we demonstrate here that activation of PKG signaling in HSC inhibits vascular tubulogenesis, migration/chemotaxis, and assembly of mature FA plaques, as assessed by vascular tubulogenesis assays and immunofluorescence localization of FA markers such as vinculin and vasodilator-stimulated phosphoprotein (VASP). To determine whether PKG inhibits FA assembly by phosphorylation of VASP at Ser-157, Ser-239, and Thr-278, we mutated these putative phosphorylation sites to alanine (VASP3A, phosphoresistant mutant) or aspartic acid (VASP3D, phosphomimetic), respectively. Data generated from these two mutants suggest that the effect of PKG on FA is independent of these three phosphorylation sites. In contrast, activation of PKG inhibits the activity of small GTPase Rac1 and its association with the effector protein IQGAP1. Moreover, PKG activation inhibits the formation of a trimeric protein complex containing Rac1, IQGAP1, and VASP. Finally, we found that expression of a constitutively active Rac1 mutant abolishes the inhibitory effects of PKG on FA formation. In summary, our data suggest that activation of PKG signaling in pericytes inhibits FA formation by inhibiting Rac1.

Preliminary investigation on the phytoplankton contribution to the mussel diet on the basis of fatty acids analysis

The composition of fatty acids was studied in the mussels collected in the Mar Grande of Taranto (northern Ionian Sea) during the four seasons. Micro-, nano- and picophytoplankton abundance, biomass and composition have been also evaluated. Fatty acids compositions were investigated for lipid biomarkers to establish the contribution of phytoplankton to the mussel diet. Saturated (SAFA) and monounsaturated fatty acids (MUFA) were the most abundant components, followed by polyunsaturated fatty acids (PUFA). The seasonal variations in the SAFAs, MUFAs and PUFAs were not significantly different during the whole study period (ANOVA,P < 0.05). The most abundant identified FAs were 16:0 (27.51–33.80% of total FAs), 14:1 (3.35–9.91% of total FAs), 18:1n9 (2.92–8.87%), 16:1 n7 (4.53– 7.61%) and 24:1n9 (0.43–8.84%). The most important PUFAs were 22:2 (2.35–3.48% of total FAs) and also 18:2n-6 (1.66–2.61%). PUFAs were characterized by low percentages of n3 and n6 FAs. Analysis of specific FA markers for diatoms (16:1n7, 20:5n3), phytoflagellates and dinoflagellates (16:0, 18:4n3) showed a negligible contribution of phytoplankton to the mussel diet.