Saturated free fatty acids and apoptosis in microvascular mesangial cells: palmitate activates pro-apoptotic signaling involving caspase 9 and mitochondrial release of endonuclease G

BACKGROUND: In type 2 diabetes, free fatty acids (FFA) accumulate in microvascular cells, but the phenotypic consequences of FFA accumulation in the microvasculature are incompletely understood. Here we investigated whether saturated FFA induce apoptosis in human microvascular mesangial cells and analyzed the signaling pathways involved. METHODS: Saturated and unsaturated FFA-albumin complexes were added to cultured human mesangial cells, after which the number of apoptotic cells were quantified and the signal transduction pathways involved were delineated. RESULTS: The saturated FFA palmitate and stearate were apoptotic unlike equivalent concentrations of the unsaturated FFA oleate and linoleate. Palmitate-induced apoptosis was potentiated by etomoxir, an inhibitor of mitochondrial β-oxidation, but was prevented by an activator of AMP-kinase, which increases fatty acid β-oxidation. Palmitate stimulated an intrinsic pathway of pro-apoptotic signaling as evidenced by increased mitochondrial release of cytochrome-c and activation of caspase 9. A caspase 9-selective inhibitor blocked caspase 3 activation but incompletely blocked apoptosis in response to palmitate, suggesting an additional caspase 9-independent pathway. Palmitate stimulated mitochondrial release of endonuclease G by a caspase 9-independent mechanism, thereby implicating endonuclease G in caspase 9-indpendent regulation of apoptosis by saturated FFA. We also observed that the unsaturated FFA oleate and linoleate prevented palmitate-induced mitochondrial release of both cytochrome-c and endonuclease G, which resulted in complete protection from palmitate-induced apoptosis. CONCLUSIONS: Taken together, these results demonstrate that palmitate stimulates apoptosis by evoking an intrinsic pathway of proapoptotic signaling and identify mitochondrial release of endonuclease G as a key step in proapoptotic signaling by saturated FFA and in the anti-apoptotic actions of unsaturated FFA

Circulating free fatty acids, insulin, and glucose during chemical stimulation of hypothalamus in rats

The aim of this study was to investigate plasma free fatty acids (FFA), insulin, and blood glucose during chemical stimulation of the lateral and ventromedial hypothalamic areas (LHA and VMH) in rats. Therefore male Wistar rats were implanted with bilateral cannulas in the LHA or the VMH and into the left and right jugular veins. Freely moving rats were then infused into the LHA and VMH with norepinephrine (NE), epinephrine (E), or acetylcholine or intravenously with NE or E. Before, during, and after the infusions, simultaneous blood samples were taken without disturbing the animals. Infusion of NE into the LHA resulted in a decrease of plasma FFA and a simultaneous increase of insulin. NE infusion in the VMH elicited an increase of plasma FFA, plasma insulin, and blood glucose. E infusion into the LHA did not lead to a change of plasma FFA, whereas insulin and glucose showed an increase. E infusion into the VMH evoked increases of plasma FFA and insulin. Peripheral administration of NE led to a sharp increase of FFA, whereas plasma insulin and blood glucose did not change. E in the periphery elicited an augmentation of plasma FFA and blood glucose and a suppression of insulin during infusion. After termination of E infusion, plasma FFA and glucose levels decreased, whereas plasma insulin showed a sharp increase. It is concluded 1) that the effects produced by administration of NE and E are dependent on hypothalamic localization and local receptor population characteristics; 2) that there are striking differences regarding the effects on the investigated blood parameters between hypothalamically infused NE and E and peripherally infused NE and E; and 3) that the LHA and VMH are able to alter plasma FFA levels independently of blood glucose and insulin levels.

Density of States FFA analysis of SU(3) lattice gauge theory at a finite density of color sources

We present a Density of States calculation with the Functional Fit Approach (DoS FFA) in SU(3) lattice gauge theory with a finite density of static color sources. The DoS FFA uses a parameterized density of states and determines the parameters of the density by fitting data from restricted Monte Carlo simulations with an analytically known function. We discuss the implementation of DoS FFA and the results for a qualitative picture of the phase diagram in a model which is a further step towards implementing DoS FFA in full QCD. We determine the curvature $\kappa$ in the $\mu$-$T$ phase diagram and find a value close to the results published for full QCD.Comment: One figure added, as well as comments on: Monte Carlo simulation, discretization effects, volume dependence, density integral. Final version to appear in Phys. Lett.

Esterification of Free Fatty Acids with Glycerol within the Biodiesel Production Framework

Companies in the field of the collection and treatment of waste cooking oils (WCO) for subsequent biodiesel production usually have to cope with high acidity oils, which cannot be directly transformed into fatty acid methyl esters due to soap production. Since glycerine is the main byproduct of biodiesel production, these high acidity oils could be esterified with the glycerine surplus to transform the free fatty acids (FFA) into triglycerides before performing the transesterification. In this work, commercial glycerol was esterified with commercial fatty acids and commercial fatty acid/lampante olive oil mixtures over tin (II) chloride. In the first set of experiments, the esterification of linoleic acid with glycerol excess from 20 to 80% molar over the stoichiometric was performed. From 20% glycerol excess, there was no improvement in FFA reduction. Using 20% glycerol excess, the performance of a biochar obtained from heavy metal-contaminated plant roots was compared to that of SnCl2. Then, the effect of the initial FFA content was assessed using different oleic acid/lampante olive oil mixtures. The results illustrated that glycerolysis was impeded at initial FFA contents lower than 10%. Finally, the glycerolysis of a WCO with 9.94% FFA was assayed, without success

Voltage-dependent Block of the Cystic Fibrosis Transmembrane Conductance Regulator Cl- Channel by Two Closely Related Arylaminobenzoates

The gene defective in cystic fibrosis encodes a Cl- channel, the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR is blocked by diphenylamine-2-carboxylate (DPC) when applied extracellularly at millimolar concentrations. We studied the block of CFTR expressed in Xenopus oocytes by DPC or by a closely related molecule, flufenamic acid (FFA). Block of whole-cell CFTR currents by bath-applied DPC or by FFA, both at 200 µM, requires several minutes to reach full effect. Blockade is voltage dependent, suggesting open-channel block: currents at positive potentials are not affected but currents at negative potentials are reduced. The binding site for both drugs senses ~40% of the electric field across the membrane, measured from the inside. In single-channel recordings from excised patches without blockers, the conductance was 8.0 ± 0.4 pS in symmetric 150 mM Cl^-. A subconductance state, measuring ~60% of the main conductance, was often observed. Bursts to the full open state lasting up to tens of seconds were uninterrupted at depolarizing membrane voltages. At hyperpolarizing voltages, bursts were interrupted by brief closures. Either DPC or FFA (50 µM) applied to the cytoplasmic or extracellular face of the channel led to an increase in flicker at V_m =-100 mV and not at V_m = +100 mV, in agreement with whole-cell experiments. DPC induced a higher frequency of flickers from the cytoplasmic side than the extracellular side. FFA produced longer closures than DPC; the FFA closed time was roughly equal (~ 1.2 ms) at -100 mV with application from either side. In cell-attached patch recordings with DPC or FFA applied to the bath, there was flickery block at V_m = -100 mV, confirming that the drugs permeate through the membrane to reach the binding site. The data are consistent with the presence of a single binding site for both drugs, reached from either end of the channel. Open-channel block by DPC or FFA may offer tools for use with site-directed mutagenesis to describe the permeation pathway

Deacidification of high acidic rubber seed oil by reesterification with glycerol

Some studies have shown that rubber seed oil (RSO) from the seed of rubber tree (hevea brasiliensis)has potential technical applications. However, the high free fatty acid (FFA) associated with the oil may be a limiting factor in its applications. In this study, crude rubber seed oil (RSO) containing 37.69% FFA was deacidified using 4.3-5.6% of glycerol. The reactions were carried out at a pressure of 20mmHg in the presence of zinc dust and zinc chloride catalysts added separately at high temperature (150 – 2500C). The progress of the reaction was monitored by determining the FFA at various time intervals.The FFA content in the RSO was reduced to 1.5% in six hours when 4.3% glycerol was used with zinc dust at a temperature of 2000C while the triglycerides content of the oil increased from 40.32% in the crude oil to 80.62% in the deacidified RSO. The study showed that this process is effective in the deacidification of RSO without loss of neutral glycerides as in alkali neutralization

Developing and testing the density of states FFA method in the SU(3) spin model

The Density of States Functional Fit Approach (DoS FFA) is a recently proposed modern density of states technique suitable for calculations in lattice field theories with a complex action problem. In this article we present an exploratory implementation of DoS FFA for the SU(3) spin system at finite chemical potential $\mu$ - an effective theory for the Polyakov loop. This model has a complex action problem similar to the one of QCD but also allows for a dual simulation in terms of worldlines where the complex action problem is solved. Thus we can compare the DoS FFA results to the reference data from the dual simulation and assess the performance of the new approach. We find that the method reproduces the observables from the dual simulation for a large range of $\mu$ values, including also phase transitions, illustrating that DoS FFA is an interesting approach for exploring phase diagrams of lattice field theories with a complex action problem.Comment: Plot, reference and comments added. Final version to appear in Nucl. Phys.

A New Test for the Absorption Mechanism of GPS Radio Sources Using Polarization Properties

We consider the use of polarization properties as a means to discriminate between Synchrotron Self-Absorption (SSA) and Free--Free Absorption (FFA) in GHz-Peaked Spectrum (GPS) sources. The polarization position angle (PA) of synchrotron radiation at high frequencies for the optically thin regime is perpendicular to the magnetic field, whereas it is parallel to the magnetic field at low frequencies for the optically thick regime. Therefore, SSA produces a change in PA of $90^{\circ}$ across the spectral peak, while FFA does not result in such a change. We analyzed polarization data from VLA observations for six GPS sources to see if such a change in PA was present. Our results indicate that there is no significant evidence for $90^{\circ}$ change in PA across the spectral peak, suggesting that FFA is more likely than SSA for low-frequency cutoffs in these sources

Eleven years of radio monitoring of the Type IIn supernova SN 1995N

We present radio observations of the optically bright Type IIn supernova SN 1995N. We observed the SN at radio wavelengths with the Very Large Array (VLA) for 11 years. We also observed it at low radio frequencies with the Giant Metrewave Radio Telescope (GMRT) at various epochs within $6.5-10$ years since explosion. Although there are indications of an early optically thick phase, most of the data are in the optically thin regime so it is difficult to distinguish between synchrotron self absorption (SSA) and free-free absorption (FFA) mechanisms. However, the information from other wavelengths indicates that the FFA is the dominant absorption process. Model fits of radio emission with the FFA give reasonable physical parameters. Making use of X-ray and optical observations, we derive the physical conditions of the shocked ejecta and the shocked CSM.Comment: 22 pages, 2 tables, 13 figures, Accepted for publication in Astrophysical Journa