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Efficient extraction of valuable products from microalgae and utilization of the residual biomass for biofuel production appear to bring economic benefits for the microalgae biorefineries. The pulsed electric field (PEF) method has been proposed as a promising pre-treatment for microalgae wet extraction. For conversion of microalgae biomass and residues, hydrothermal liquefaction (HTL) is considered a suitable method. Here, a combination of PEF-assisted valuables extraction of microalgae and HTL of the residual biomass is studied for the first time. The microalgae Auxenochlorella protothecoides, Chlorella vulgaris, and Scenedesmus almeriensis were cultivated, harvested, treated by PEF, and then subjected to lipid extraction, protein extraction, or extraction of amino acids after enzymatic protein hydrolysis. The residual biomass obtained from PEF-assisted valuables extraction was subjected to HTL in micro autoclaves at a temperature of 350 °C and pressure of 25 MPa for 15 min holding time. Product yields and analytical results obtained by elemental content measurement, 1H-nuclear magnetic resonance spectroscopy, Fourier-transform infrared spectroscopy, and gel permeation chromatography show that PEF alone has no significant direct influence on microalgae HTL. The harsh HTL conditions decisively determine the product yields and biocrude quality. However, PEF enhances the lipid extraction yield from 4 wt% to 33 wt%. Consequently, the biocrude yield decreases from 58 wt% to 43.2 wt%. Moreover, PEF boosts the protein extraction yield from almost zero to 41.6 wt% of the total protein, resulting in an increased biocrude yield by about 2 wt%. Finally, PEF accelerates the formation of amino acids by enzymatic hydrolysis, improving the extraction efficiency up to 150% in the first 60 min of the extraction. The extracted residue promises to produce a biocrude yield higher by 6 wt% and a better biocrude quality with lower nitrogen content. An overall mass balance of PEF-assisted valuables extraction and HTL is presented.

Pulsed electric field (PEF) treatment has so far been considered as ineffective for protein extraction, but the parameters that influence protein extraction efficiency have not been identified fully. Using Chlorella vulgaris in this study, we show that up to half of the proteins present in the cell can be extracted via PEF treatment and an incubation step for up to 24 h. The extraction efficiency is highly influenced by biomass concentration during the incubation step and the incubation temperature: in our experiments we were able to extract half of the proteins present in the cells at dilute concentrations (5 mg·ml−1), but this efficiency decreases with increasing biomass concentration, indicating that diffusion gradients play a role in the protein release. However, we also show that not only diffusion plays a role, but also a biological process that requires suitable conditions. We observed that PEF treated cells undergo programmed cell death, indicated by DNA laddering. Furthermore cold and hot temperatures as well as protease inhibition impair the release of proteins. In Western blots we can observe that some proteins are not released when the activity of proteases is blocked. We therefore conclude that protein release is facilitated by an enzyme-driven process that occurs after PEF-triggered cell death, and that this process is prone to extreme conditions.

Objectives This study sought to test the hypothesis that inducing an autoimmune response against the cardiac sodium channel (NaV1.5) induces arrhythmias. Background Sporadic evidence supports the concept that autoantibodies may cause cardiac arrhythmias but substantial experimental investigations using in vivo models have been lacking to date. The NaV1.5 is essential for cardiac impulse propagation and its dysfunction has been linked to conduction disease. Methods Rats were immunized with a peptide sequence derived from the third extracellular loop of the first domain of NaV1.5. After 28 days, we evaluated in vivo both the electrical and mechanical parameters of cardiac function. Histopathology, myocardial gene and protein expression were assessed. Whole-cell patch-clamp was used to measure sodium current (INa) density in isolated cardiomyocytes. Results NaV1.5-immunized rats had high titers of autoantibodies against NaV1.5. On ECG recording, NaV1.5-immunized animals showed significantly prolonged PR-intervals. During Holter ECG-monitoring we observed repeated prolonged episodes of third-degree atrioventricular and sinoatrial block in every NaV1.5-immunized animal, but not in controls. Immunization had no effect on cardiac function. In comparison to controls, myocardial NaV1.5 mRNA and protein levels were decreased in immunized rats. INa density was reduced in cardiomyocytes incubated with sera from NaV1.5-immunized rats and from patients with idiopathic atrioventricular block (AVB) in comparison to sera from respective controls. In patients with idiopathic AVB, we observed autoantibodies against NaV1.5 that were absent in sera from healthy controls. Conclusions Provocation of an autoimmune response against NaV1.5 induces conductance defects probably caused by a reduced expression level and an inhibition of NaV1.5 by autoantibodies, resulting in decreased INa.

Kir2.x channels form the molecular basis of cardiac I(K1) current and play a major role in cardiac electrophysiology. However, there is a substantial lack of selective Kir2 antagonists. We found the β(3)-adrenoceptor antagonist SR59230A to be an inhibitor of Kir2.x channels. Therefore, we characterized the effects of SR59230A on Kir2.x and other relevant cardiac potassium channels. Cloned channels were expressed in the Xenopus oocyte expression system and measured with the double-microelectrode voltage clamp technique. SR59230A inhibited homomeric Kir2.1 channels with an IC(50) of 33μM. Homomeric Kir2.2 and Kir2.3 channels and Kir2.x heteromers were also inhibited by SR59230A with similar potency. In contrast, no relevant inhibitory effects of SR59230A were found in cardiac Kv1.5, Kv4.3 and KvLQT1/minK channels. In hERG channels, SR59230A only induced a weak inhibition at a high concentration. These findings establish SR59230A as a novel inhibitor of Kir2.1-2.3 channels with a favorable profile with respect to additional effects on other cardiac repolarizing potassium channels.

There is genetic and epidemiological evidence demonstrating the association between Lp(a) levels and cardiovascular disease. While evolocumab reduces Lp(a) by 25-30%, the impact of PCSK9 inhibition on plaque at different Lp(a) levels is unknown. GLAGOV compared the effects of the PCSK9 inhibitor

INHIBITION OF HETEROMERIC KIR2.X CHANNELS BY ALPHA1A ADRENERGIC RECEPTORS: CRUCIAL ROLE OF PROTEIN KINASE C DEPENDENT REGULATION OF KIR2.3 SUBUNITS Claudia Kiesecker, MS, Edgar Zitron, Daniel Scherer, Sonja Lueck, Ramona Bloehs, Eberhard P. Scholz, Sven Kathofer, MD, Dierk Thomas, MD, Volker A. Kreye, MD, Johann Kiehn, MD, Wolfgang Schols, MD, Hugo A. Katus, MD and Christoph A. Karle, MD. Medical University Hospital Heidelberg, Heidelberg, Germany and University of Heidelberg, Heidelberg, Germany.