The formation of condensed (compacted) protein phases is associated with a wide range of human disorders, such as eye cataracts, amyotrophic lateral sclerosis, sickle cell anaemia and Alzheimer's ...disease. However, condensed protein phases have their uses: as crystals, they are harnessed by structural biologists to elucidate protein structures, or are used as delivery vehicles for pharmaceutical applications. The physiochemical properties of crystals can vary substantially between different forms or structures ('polymorphs') of the same macromolecule, and dictate their usability in a scientific or industrial context. To gain control over an emerging polymorph, one needs a molecular-level understanding of the pathways that lead to the various macroscopic states and of the mechanisms that govern pathway selection. However, it is still not clear how the embryonic seeds of a macromolecular phase are formed, or how these nuclei affect polymorph selection. Here we use time-resolved cryo-transmission electron microscopy to image the nucleation of crystals of the protein glucose isomerase, and to uncover at molecular resolution the nucleation pathways that lead to two crystalline states and one gelled state. We show that polymorph selection takes place at the earliest stages of structure formation and is based on specific building blocks for each space group. Moreover, we demonstrate control over the system by selectively forming desired polymorphs through site-directed mutagenesis, specifically tuning intermolecular bonding or gel seeding. Our results differ from the present picture of protein nucleation, in that we do not identify a metastable dense liquid as the precursor to the crystalline state. Rather, we observe nucleation events that are driven by oriented attachments between subcritical clusters that already exhibit a degree of crystallinity. These insights suggest ways of controlling macromolecular phase transitions, aiding the development of protein-based drug-delivery systems and macromolecular crystallography.
Self-assembly of proteins holds great promise for the bottom-up design and production of synthetic biomaterials. In conventional approaches, designer proteins are pre-programmed with specific ...recognition sites that drive the association process towards a desired organized state. Although proven effective, this approach poses restrictions on the complexity and material properties of the end-state. An alternative, hierarchical approach that has found wide adoption for inorganic systems, relies on the production of crystalline nanoparticles that become the building blocks of a next-level assembly process driven by oriented attachment (OA). As it stands, OA has not yet been observed for protein systems. Here we employ cryo-transmission electron microscopy (cryoEM) in the high nucleation rate limit of protein crystals and map the self-assembly route at molecular resolution. We observe the initial formation of facetted nanocrystals that merge lattices by means of OA alignment well before contact is made, satisfying non-trivial symmetry rules in the process. As these nanocrystalline assemblies grow larger we witness imperfect docking events leading to oriented aggregation into mesocrystalline assemblies. These observations highlight the underappreciated role of the interaction between crystalline nuclei, and the impact of OA on the crystallization process of proteins.
Abstract Synthetic micro/nanomotors have been extensively exploited over the past decade to achieve active transportation. This interest is a result of their broad range of potential applications, ...from environmental remediation to nanomedicine. Nevertheless, it still remains a challenge to build a fast-moving biodegradable polymeric nanomotor. Here we present a light-propelled nanomotor by introducing gold nanoparticles (Au NP) onto biodegradable bowl-shaped polymersomes (stomatocytes) via electrostatic and hydrogen bond interactions. These biodegradable nanomotors show controllable motion and remarkable velocities of up to 125 μm s −1 . This unique behavior is explained via a thorough three-dimensional characterization of the nanomotor, particularly the size and the spatial distribution of Au NP, with cryogenic transmission electron microscopy (cryo-TEM) and cryo-electron tomography (cryo-ET). Our in-depth quantitative 3D analysis reveals that the motile features of these nanomotors are caused by the nonuniform distribution of Au NPs on the outer surface of the stomatocyte along the z-axial direction. Their excellent motile features are exploited for active cargo delivery into living cells. This study provides a new approach to develop robust, biodegradable soft nanomotors with application potential in biomedicine.
Dynamic Reweighting of Auditory Modulation Filters Joosten, Eva R M; Shamma, Shihab A; Lorenzi, Christian ...
PLOS computational biology/PLoS computational biology,
07/2016, Letnik:
12, Številka:
7
Journal Article
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Sound waveforms convey information largely via amplitude modulations (AM). A large body of experimental evidence has provided support for a modulation (bandpass) filterbank. Details of this model ...have varied over time partly reflecting different experimental conditions and diverse datasets from distinct task strategies, contributing uncertainty to the bandwidth measurements and leaving important issues unresolved. We adopt here a solely data-driven measurement approach in which we first demonstrate how different models can be subsumed within a common 'cascade' framework, and then proceed to characterize the cascade via system identification analysis using a single stimulus/task specification and hence stable task rules largely unconstrained by any model or parameters. Observers were required to detect a brief change in level superimposed onto random level changes that served as AM noise; the relationship between trial-by-trial noisy fluctuations and corresponding human responses enables targeted identification of distinct cascade elements. The resulting measurements exhibit a dynamic complex picture in which human perception of auditory modulations appears adaptive in nature, evolving from an initial lowpass to bandpass modes (with broad tuning, Q∼1) following repeated stimulus exposure.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Swimming events in city canals are gaining popularity in the Netherlands, even though canal water is usually not officially designated for recreational use. Knowledge regarding the risk of infection ...after swimming in canals is limited. An outbreak was reported in 2015 following a canal swimming event in Utrecht, the Netherlands. Local governments were concerned about the health risks of such events. In order to assess the safety of canal swimming, the Public Health Service (PHS) prospectively investigated two city canal swimming events in 2015. In 2016, we repeated this study, aiming to prospectively determine the risks of infection during two urban swimming events, the Utrecht SingelSwim 2016 (USS) and the Amsterdam City Swim 2016 (ACS).
We sent online questionnaires to 271 USS participants and 2697 ACS participants, concerning personal characteristics, symptoms, and exposure. Participants were asked to forward the questionnaire to three relatives, i.e., non-exposed. We analyzed water samples from the USS venue taken during the event, as well as stool samples of USS participants with acute gastrointestinal illness (AGI). AGI was defined as diarrhea and/or vomiting within seven days after the event. We calculated adjusted risk ratios (RR) for AGI in the exposed group compared with non-exposed respondents, using binomial regression models.
The questionnaire was returned by 160 USS participants (exposed) (59%) and 40 non-exposed relatives. Five percent of the exposed (n = 17) and 3% of non-exposed (n = 1) reported AGI (RR = 1.69; 95% CI: 0.23-12.46). Norovirus genogroup II was detected in two of six USS water samples and in none of the three stool samples. In one of three stool samples, rotavirus was detected. The questionnaire was returned by 1169 ACS participants (exposed) (43%) and 410 non-exposed relatives. Six percent of the exposed (n = 71) and 1% of non-exposed (n = 5) reported AGI (RR 4.86; 95% CI: 1.98-11.97).
Results of the ACS event showed a higher risk for AGI among the exposed, indicating that participants of events in urban canals in the Netherlands could be at a higher risk for AGI than those not participating. The inconclusive results from the USS are likely due to the small sample size. Swimming in non-monitored open water can bring health risks and more knowledge about environmental and human risk factors helps reduce the risk by being able to more specifically advise organizations and governments.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
We investigate the photocatalytic performance of composites prepared in a one-step process by liquid-phase exfoliation of graphite in the presence of TiO2 nanoparticles (NPs) at atmospheric pressure ...and in water, without heating or adding any surfactant, and starting from low-cost commercial reagents. These show enhanced photocatalytic activity, degrading up to 40% more pollutants with respect to the starting TiO2-NPs, in the case of a model dye target, and up to 70% more pollutants in the case of nitrogen oxides. In order to understand the photo-physical mechanisms underlying this enhancement, we investigate the photo-generation of reactive species (trapped holes and electrons) by ultrafast transient absorption spectroscopy. We observe an electron transfer process from TiO2 to the graphite flakes within the first picoseconds of the relaxation dynamics, which causes the decrease of the charge recombination rate, and increases the efficiency of the reactive species photo-production.
Abstract Various processes might explain the progression from casual to compulsive drug use underlying the development of drug addiction. Two of these, accelerated stimulus–response (S–R) habit ...learning and augmented assignment of motivational value to reinforcers, could be mediated via neuroadaptations associated with long-lasting sensitization to psychostimulant drugs, i.e. augmented dopaminergic neurotransmission in the striatum. Here, we tested the hypothesis that both processes, which are often regarded as mutually exclusive alternatives, are present in amphetamine-sensitized rats. Amphetamine-sensitized rats showed increased responding for food under a random ratio schedule of reinforcement, indicating increased incentive motivational value of food. In addition, satiety-specific devaluation experiments under a random interval schedule of reinforcement showed that amphetamine-sensitized animals exhibit accelerated development of S–R habits. These data show that both habit formation and motivational value of reinforcers are augmented in amphetamine-sensitized rats, and suggest that the task demands determine which behavioral alteration is most prominently expressed.
Polyamines play a major role in biosilicification reactions in diatoms and sponges. While the effects of polyamines on silicic acid oligomerization and precipitation are well known, the impact of ...polyamines chain length on silica particle growth is unclear. We studied the effects of polyamine chain length on silica particle growth and condensation in a known, simple, and salt-free biphasic reaction system; with tetraethyl orthosilicate as organic phase and polyamine dissolved in the aqueous phase. The particles at various growth stages were characterized by Cryo- Transmission Electron Microscopy, Scanning Electron Microscopy, Thermogravimetric Analysis, Zeta Potential, and solid-state NMR analysis. Polyamines were found co-localized within silica particles and the particle diameter increased with an increase in polyamine chain length, whereas silica condensation showed the opposite trend. Particle growth is proposed to progress via a coacervate intermediate while the final particles have a core shell structure with an amine-rich core and silica-rich shell. The results presented in this paper would of interest for researchers working in the field of bioinspired materials.
The mineralization of collagen via synthetic procedures has been extensively investigated for hydroxyapatite as well as for silica and calcium carbonate. From a fundamental point of view, it is ...interesting to investigate whether collagen could serve as a generic mineralization template for other minerals, like iron oxides. Here, bio-inspired coprecipitation reaction, generally leading to the formation of magnetite, is used to mineralize collagen with iron hydroxides. Platelet-shaped green rust crystals form outside the collagen matrix, while inside the collagen, nanoparticles with a size of 2.6 nm are formed, which are hypothesized to be iron (III) hydroxide. Mineralization with nanoparticles inside the collagen solely occurs in the presence of poly(aspartic acid) (pAsp). In the absence of pAsp, magnetite particles are formed around the collagen. Time-resolved cryo-TEM shows that during the coprecipitation reaction, initially a beam-sensitive phase is formed, possibly an Fe3+–pAsp complex. This beam-sensitive phase transforms into nanoparticles. In a later stage, sheet-like crystals are also found. After 48 h of mineralization, ordering of the nanoparticles around one of the collagen sub-bands (the a-band) is observed. This is very similar to the collagen–hydroxyapatite system, indicating that mineralization with iron hydroxides inside collagen is possible and proceeds via a similar mechanism as hydroxyapatite mineralization.
Liquid‐Phase (Scanning) Transmission Electron Microscopy (LP‐(S)TEM) has become an essential technique to monitor nanoscale materials processes in liquids in real‐time. Due to the pressure difference ...between the liquid and the microscope vacuum, bending of the silicon nitride (SiNx) membrane windows generally occurs. This causes a spatially varying liquid layer thickness that makes interpretation of LP‐(S)TEM results difficult due to a locally varying achievable resolution and diffusion limitations. To mediate these difficulties, it is shown: 1) how to quantitatively map liquid layer thickness for any liquid at less than 0.01 e− Å−2 total dose; 2) how to dynamically modulate the liquid thickness by tuning the internal pressure in the liquid cell, co‐determined by the Laplace pressure and the external pressure. It is demonstrated that reproducible inward bulging of the window membranes can be realized, leading to an ultra‐thin liquid layer in the central window area for high‐resolution imaging. Furthermore, it is shown that the liquid thickness can be dynamically altered in a programmed way, thereby potentially overcoming the diffusion limitations towards achieving bulk solution conditions. The presented approaches provide essential ways to measure and dynamically adjust liquid thickness in LP‐(S)TEM experiments, enabling new experiment designs and better control of solution chemistry.
Real‐time monitoring of nanoscale material processes in liquids by liquid‐phase (scanning) transmission electron microscopy is advanced by rapid dynamic control and quantification of the liquid layer thickness. These methods enable performing the largest part of the experiment using a thick liquid layer to avoid confinement effects and adjusting to a thin liquid layer whenever needed for imaging at higher resolution.
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