Regenerative NanoOctopus Based on Multivalent-Aptamer-Functionalized Magnetic Microparticles for Effective Cell Capture in Whole Blood.

Isolation of specific rare cell subtypes from whole blood is critical in cellular analysis and important in basic and clinical research. Traditional immunomagnetic cell capture suffers from suboptimal sensitivity, specificity, and time- and cost-effectiveness. Mimicking the features of octopuses, a device termed a "NanoOctopus" was developed for cancer cell isolation in whole blood. The device consists of long multimerized aptamer DNA strands, or tentacle DNA, immobilized on magnetic microparticle surfaces. Their ultrahigh sensitivity and specificity are attributed to multivalent binding of the tentacle DNA to cell receptors without steric hindrance. The simple, quick, and noninvasive capture and release of the target cells allows for extensive downstream cellular and molecular analysis, and the time- and cost-effectiveness of fabrication and regeneration of the devices makes them attractive for industrial manufacture.

Cryospectroscopy Studies of Intact Light-Harvesting Antennas Reveal Empirical Electronic Energy Transitions in Two Cyanobacteria Species.

Understanding of electronic energy transition (EET) mechanisms from the light-harvesting unit to the reaction center in a natural system has been limited by (a) the use of conventional transient time-resolved spectroscopy at room temperature, which result in high signal-to-noise ratio and (b) examining extracts instead of intact light-harvesting units. Here, we report previously unknown differences and new insight in EET of two cyanobacteria species, Acaryochloris marina and Thermosynechoccocus vulcanus, which have been found only after using UV-vis, hole-burning, and fluorescence spectroscopy at ultralow temperature and examining their intact light-harvesting unit, phycobilisomes (PBS). Although the exciton formation is similarly induced by photoexcitation of chromophore assemblies in phycocyanin (PC) and allophycocyanin (APC) in PBSs of both species, the EET mechanisms are totally different, being adiabatic in A. marina and nonadiabatic in T. vulcanus. The PBS of A. marina has only one APC trimer and energy transfer is through coupling of α84 in APC with ß84 in adjacent PC. In T. vulcanus, the PBS has three components: coupling between APC core and the entire PC rod and couplings of ß-ß18 and of LCM to ß in the adjacent APC-like trimer. A total of 80% of the excitation energy is trapped in the coupling ß-ß18 and regulates the flow of energy from the high- to low-level terminal electronic transition emitter ß-LCM. All these details cannot be observed at room temperature and in extracted units.

Energy Transfer Kinetics in Photosynthesis as an Inspiration for Improving Organic Solar Cells.

Clues to designing highly efficient organic solar cells may lie in understanding the architecture of light-harvesting systems and exciton energy transfer (EET) processes in very efficient photosynthetic organisms. Here, we compare the kinetics of excitation energy tunnelling from the intact phycobilisome (PBS) light-harvesting antenna system to the reaction center in photosystem II in intact cells of the cyanobacterium Acaryochloris marina with the charge transfer after conversion of photons into photocurrent in vertically aligned carbon nanotube (va-CNT) organic solar cells with poly(3-hexyl)thiophene (P3HT) as the pigment. We find that the kinetics in electron hole creation following excitation at 600 nm in both PBS and va-CNT solar cells to be 450 and 500 fs, respectively. The EET process has a 3 and 14 ps pathway in the PBS, while in va-CNT solar cell devices, the charge trapping in the CNT takes 11 and 258 ps. We show that the main hindrance to efficiency of va-CNT organic solar cells is the slow migration of the charges after exciton formation.

Disagreement Between the Structure of the dTpT Thymine Pair Determined by NMR and Molecular Dynamics Simulations Using Amber 14 Force Fields.

We report a disagreement between the predicted structures of the dTpT thymine pair (thymidylyl(3' → 5')thymidine) using nuclear magnetic resonance (NMR) spectroscopy and molecular dynamics (MD) simulations using the AMBER ff14SB and ff14 + ε/ζOL1 + χOL4 force fields for DNA. The NMR structure was determined using NOE couplings to thymine's H6 and J(HH) couplings between sugar protons. The MD simulation used replica exchange methods to produce converged statistics in a 500 ns trajectory. NMR data indicate that both thymine nucleotides in the pair display an anti conformation of B-DNA, while the MD simulations predict a structure in which the 5'-thymine is flipped into a syn conformation and the 3'-thymine is in an anti conformation. The syn conformation of the 5'-thymine predicted by MD appears by a ∼ 180-deg flip of the glycosidic angle in comparison to the B-form anti structure. Differences in the distortion of the sugar pucker between 5'-thymine and 3'-thymine further highlighted the surprisingly different conformation of the 5'- and 3'-ends. While both MD and NMR indicate the deoxyribose sugars to be primarily in the 2'-endo conformation typical of B-form DNA, the MD simulations predict a more twisted conformation (2'-endo/1'-exo) for the 5'-sugar and significant flexibility of C3' of the 3'-sugar. We conclude that the current AMBER force field does not accurately predict the conformation of single-stranded thymine, in agreement with previous work investigating single-stranded DNA.

Adsorption of doxorubicin on citrate-capped gold nanoparticles: insights into engineering potent chemotherapeutic delivery systems.

Gold nanomaterials have received great interest for their use in cancer theranostic applications over the past two decades. Many gold nanoparticle-based drug delivery system designs rely on adsorbed ligands such as DNA or cleavable linkers to load therapeutic cargo. The heightened research interest was recently demonstrated in the simple design of nanoparticle-drug conjugates wherein drug molecules are directly adsorbed onto the as-synthesized nanoparticle surface. The potent chemotherapeutic, doxorubicin often serves as a model drug for gold nanoparticle-based delivery platforms; however, the specific interaction facilitating adsorption in this system remains understudied. Here, for the first time, we propose empirical and theoretical evidence suggestive of the main adsorption process where (1) hydrophobic forces drive doxorubicin towards the gold nanoparticle surface before (2) cation-π interactions and gold-carbonyl coordination between the drug molecule and the cations on AuNP surface facilitate DOX adsorption. In addition, biologically relevant compounds, such as serum albumin and glutathione, were shown to enhance desorption of loaded drug molecules from AuNP at physiologically relevant concentrations, providing insight into the drug release and in vivo stability of such drug conjugates.

Low absorption state of phycocyanin from Acaryochloris marina antenna system: on the interplay between ionic strength and excitonic coupling.

This paper studies the excitonic factor in the excited state energy transfer of phycobilisome (PBS) by using a polarized time-resolved pump-probe and by changing the ionic strength of the cofactors' medium in the PBS of Acaryochloris marina (A. marina). As a result, the interplay between the surrounding medium and the closely excited adjacent cofactors is shown to be a negligible factor of the excitonic decay kinetics at 618 nm of the phycocyanin (PC), while it appears as a driving factor of an increase in excitonic delocalization at 630 nm. The obtained anisotropy values are consistent with the contribution of ionic strength in the excitonic mechanism in PBS. These values were 0.38 in high ionic strength and 0.4 in low ionic strength at 618 nm, and 0.52 in high ionic strength and 0.4 in low ionic strength at 630-635 nm. The anisotropy value of 0.52 in high phosphate is similar at 630 nm and 635 nm, which is consistent with an excitonic delocalization band at 635 nm. The 635 nm band is suggested to show the true low energy level of PC in A. marina PBS. The anisotropy decay kinetic at 630 nm suggests that the excited state population of PC is not all equilibrated in 3 ps because of the existence of the 10 ps decay kinetic component. The presence of the slow kinetic decay component in high, and low ionic strength, is consistent with a 10 and 14 ps energy transfer pathway, while the 450 fs kinetic decay component is consistent with the presence of an additional excitation energy transfer pathway between adjacent α84 and ß84. Furthermore, the 450 fs decay kinetic is suggested to be trapped in the trimer, while the 400 fs decay kinetic rules out an excitonic flow from low energy level PC to allophycoyanin. This excitonic flow may occur between ß84 in adjacent trimers, towards the low energy state of the PBS rod.

Excitation energy transfer in intact cells and in the phycobiliprotein antennae of the chlorophyll d containing cyanobacterium Acaryochloris marina.

The cyanobacterium Acaryochloris marina is unique because it mainly contains Chlorophyll d (Chl d) in the core complexes of PS I and PS II instead of the usually dominant Chl a. Furthermore, its light harvesting system has a structure also different from other cyanobacteria. It has both, a membrane-internal chlorophyll containing antenna and a membrane-external phycobiliprotein (PBP) complex. The first one binds Chl d and is structurally analogous to CP43. The latter one has a rod-like structure consisting of three phycocyanin (PC) homohexamers and one heterohexamer containing PC and allophycocyanin (APC). In this paper, we give an overview on the investigations of excitation energy transfer (EET) in this PBP-light-harvesting system and of charge separation in the photosystem II (PS II) reaction center of A. marina performed at the Technische Universität Berlin. Due to the unique structure of the PBP antenna in A. marina, this EET occurs on a much shorter overall time scale than in other cyanobacteria. We also briefly discuss the question of the pigment composition in the reaction center (RC) of PS II and the nature of the primary donor of the PS II RC.