Ionic selectivity in L-type calcium channels by electrostatics and hard-core repulsion

A physical model of selective “ion binding” in the L-type calcium channel is constructed, and consequences of the model are compared with experimental data. This reduced model treats only ions and the carboxylate oxygens of the EEEE locus explicitly and restricts interactions to hard-core repulsion and ion–ion and ion–dielectric electrostatic forces. The structural atoms provide a flexible environment for passing cations, thus resulting in a self-organized induced-fit model of the selectivity filter. Experimental conditions involving binary mixtures of alkali and/or alkaline earth metal ions are computed using equilibrium Monte Carlo simulations in the grand canonical ensemble. The model pore rejects alkali metal ions in the presence of biological concentrations of Ca2+ and predicts the blockade of alkali metal ion currents by micromolar Ca2+. Conductance patterns observed in varied mixtures containing Na+ and Li+, or Ba2+ and Ca2+, are predicted. Ca2+ is substantially more potent in blocking Na+ current than Ba2+. In apparent contrast to experiments using buffered Ca2+ solutions, the predicted potency of Ca2+ in blocking alkali metal ion currents depends on the species and concentration of the alkali metal ion, as is expected if these ions compete with Ca2+ for the pore. These experiments depend on the problematic estimation of Ca2+ activity in solutions buffered for Ca2+ and pH in a varying background of bulk salt. Simulations of Ca2+ distribution with the model pore bathed in solutions containing a varied amount of Li+ reveal a “barrier and well” pattern. The entry/exit barrier for Ca2+ is strongly modulated by the Li+ concentration of the bath, suggesting a physical explanation for observed kinetic phenomena. Our simulations show that the selectivity of L-type calcium channels can arise from an interplay of electrostatic and hard-core repulsion forces among ions and a few crucial channel atoms. The reduced system selects for the cation that delivers the largest charge in the smallest ion volume

The Effect of Dichloromethane on Product Separation during Continuous Hydrothermal Liquefaction of Chlorella vulgaris and Aqueous Product Recycling for Algae Cultivation

Dichloromethane (DCM) is a solvent commonly used in laboratories for microalgae hydrothermal liquefaction (HTL) product separation. The addition of DCM would lead to an “overestimation effect” of biocrude yield and diminish biocrude quality. However, it is currently not clear to what extent this overestimation effect will impact a continuous HTL process. In this study, Chlorella vulgaris microalgae was processed in a continuous stirred tank reactor at different temperatures (300, 325, 350, 375, and 400 °C) at 24 MPa for 15 min holding time. Two separation methods were applied to investigate the effect of using DCM in a cHTL product separation procedure in terms of product yield, biocrude elemental content, and aqueous product (AP) composition. Subsequently, the feasibility of reusing AP for algae cultivation has been evaluated. Results suggest that 350 °C is the optimal temperature for cHTL operation, leading to the highest biocrude yield, and an average increase in biocrude yield of 9 wt % was achieved when using DCM in cHTL product separation. Within the temperature range investigated, an average biocrude yield estimation can be proposed by yield$_{non-DCM}$ ≈ 0.818 × yield$_{DCM}$. The AP has been characterized by total organic carbon and total nitrogen, high-performance liquid chromatography, and inductively coupled plasma optical emission spectroscopy. Results show that at 350–375 °C more nitrogen and other ions were directed into the AP, which could be advantageous in nutrient recovery. With the help of optical density testing, algae was shown to exhibit a better growth using AP with activated carbon absorption purification treatment as compared to the standard medium. The recovery of water and nutrients from the HTL-AP could improve the economics of a microalgae biorefinery process

Screening and Optimization of Microalgae Biomass and Plastic Material Coprocessing by Hydrothermal Liquefaction

In the past decade, microalgae biomass has been attracting considerable interest in valuable biocomponents and biofuel production. Meanwhile, plastic waste handling has become one of the most pressing global environmental concerns. Coprocessing of plastic waste and biomass has previously been reported to produce good quality fuel oil and high-value chemicals. In this study, we examined a coliquefaction process (co-HTL) of 2 microalgae, Chlorella vulgaris (Cv) and Nannochloropsis gaditana (Ng), with nine types of common plastics. In a first step, the co-HTL process was conducted in microautoclave reactors with a fixed algae/plastic mass ratio (50:50) at a temperature of 350 °C and a pressure of 16 MPa for a holding time of 15 min. Among the different types of plastics, positive synergistic effects between polycarbonate (PC), polystyrene (PS), and microalgae have been observed: (1) Plastics showed greater decomposition. (2) HTL crude oil yields were increased. Ng algae exhibits a higher interaction ability with plastics. Then, PC and PS were coprocessed with Ng algae using the response surface methodology to optimize the effects of temperature (300–400 °C), algae/plastic mass ratio (20:80–80:20), and holding time (5–45 min) on HTL crude oil yield. Software-based data analysis of the co-HTL experiments were conducted, and the optimal parameters were proposed, which were verified by the experiment results; Ng+PC (20:80 wt %) exhibits the highest crude oil yield of 67.2% at 300 °C with a 5 min holding time, while Ng+PS (80:20 wt %) generates 51.4 wt % crude oil yield at 400 °C and a 25 min holding time. Finally, the analytical results of elemental analysis, FTIR, 1H NMR, GPC, GC-MS, and TGA on the crude oil produced from pure microalgae HTL and co-HTL were compared, indicating that Ng+PC crude oil is more suitable for aromatic chemicals production and Ng+PS crude oil could be more favorable for biofuel applications

Effects of multiple occupancy and inter-particle interactions on selective transport through narrow channels: theory versus experiment

Many biological and artificial transport channels function without direct input of metabolic energy during a transport event and without structural rearrangements involving transitions from a 'closed' to an 'open' state. Nevertheless, such channels are able to maintain efficient and selective transport. It has been proposed that attractive interactions between the transported molecules and the channel can increase the transport efficiency and that the selectivity of such channels can be based on the strength of the interaction of the specifically transported molecules with the channel. Herein, we study the transport through narrow channels in a framework of a general kinetic theory, which naturally incorporates multi-particle occupancy of the channel and non-single-file transport. We study how the transport efficiency and the probability of translocation through the channel are affected by inter-particle interactions in the confined space inside the channel, and establish conditions for selective transport. We compare the predictions of the model with the available experimental data - and find good semi-quantitative agreement. Finally, we discuss applications of the theory to the design of artificial nano-molecular sieves.Comment: 27 pages, 6 figures, 1 Appendix, in press in Biophysical Journa

Intrafraction motion of the prostate during an IMRT session: a fiducial-based 3D measurement with Cone-beam CT

Background: Image-guidance systems allow accurate interfractional repositioning of IMRT treatments, however, these may require up to 15 minutes. Therefore intrafraction motion might have an impact on treatment precision. 3D geometric data regarding intrafraction prostate motion are rare; we therefore assessed its magnitude with pre- and post-treatment fiducial-based imaging with cone-beam-CT (CBCT). Methods: 39 IMRT fractions in 5 prostate cancer patients after (125)I-seed implantation were evaluated. Patient position was corrected based on the (125)I-seeds after pre-treatment CBCT. Immediately after treatment delivery, a second CBCT was performed. Differences in bone- and fiducial position were measured by seed-based grey-value matching. Results: Fraction time was 13.6 +/- 1.6 minutes. Median overall displacement vector length of (125)Iseeds was 3 mm (M = 3 mm, Sigma = 0.9 mm, sigma = 1.7 mm; M: group systematic error, Sigma: SD of systematic error, sigma: SD of random error). Median displacement vector of bony structures was 1.84 mm (M = 2.9 mm, Sigma = 1 mm, sigma = 3.2 mm). Median displacement vector length of the prostate relative to bony structures was 1.9 mm (M = 3 mm, Sigma = 1.3 mm, sigma = 2.6 mm). Conclusion: a) Overall displacement vector length during an IMRT session is < 3 mm. b) Positioning devices reducing intrafraction bony displacements can further reduce overall intrafraction motion. c) Intrafraction prostate motion relative to bony structures is < 2 mm and may be further reduced by institutional protocols and reduction of IMRT duration

A Structural Model of the Pore-Forming Region of the Skeletal Muscle Ryanodine Receptor (RyR1)

Ryanodine receptors (RyRs) are ion channels that regulate muscle contraction by releasing calcium ions from intracellular stores into the cytoplasm. Mutations in skeletal muscle RyR (RyR1) give rise to congenital diseases such as central core disease. The absence of high-resolution structures of RyR1 has limited our understanding of channel function and disease mechanisms at the molecular level. Here, we report a structural model of the pore-forming region of RyR1. Molecular dynamics simulations show high ion binding to putative pore residues D4899, E4900, D4938, and D4945, which are experimentally known to be critical for channel conductance and selectivity. We also observe preferential localization of Ca2+ over K+ in the selectivity filter of RyR1. Simulations of RyR1-D4899Q mutant show a loss of preference to Ca2+ in the selectivity filter as seen experimentally. Electrophysiological experiments on a central core disease mutant, RyR1-G4898R, show constitutively open channels that conduct K+ but not Ca2+. Our simulations with G4898R likewise show a decrease in the preference of Ca2+ over K+ in the selectivity filter. Together, the computational and experimental results shed light on ion conductance and selectivity of RyR1 at an atomistic level