The assembly platform FimD is required to obtain the most stable quaternary structure of type 1 pili.

Type 1 pili are important virulence factors of uropathogenic Escherichia coli that mediate bacterial attachment to epithelial cells in the urinary tract. The pilus rod is comprised of thousands of copies of the main structural subunit FimA and is assembled in vivo by the assembly platform FimD. Although type 1 pilus rods can self-assemble from FimA in vitro, this reaction is slower and produces structures with lower kinetic stability against denaturants compared to in vivo-assembled rods. Our study reveals that FimD-catalysed in vitro-assembled type 1 pilus rods attain a similar stability as pilus rods assembled in vivo. Employing structural, biophysical and biochemical analyses, we show that in vitro assembly reactions lacking FimD produce pilus rods with structural defects, reducing their stability against dissociation. Overall, our results indicate that FimD is not only required for the catalysis of pilus assembly, but also to control the assembly of the most stable quaternary structure.

Photostability of sennosides and their aglycones in solution.

INTRODUCTION: Sennosides are the main active constituents of the dried leaves and/or pods of Senna alexandrina Mill. that are used as laxatives. A hypothesis is that aglycones are formed during the degradation of sennosides. However, it is unknown, whether this happens under visible light exposure and how photosensitive sennosides behave in solution. OBJECTIVES: Pure anthraquinone glycosides were tested on their behaviour during sample preparation in the lab under visible light exposure in dependence on the instability of the solvent. MATERIALS AND METHODS: Samples before and after exposure were analysed using UHPLC with UV/Vis and MS detection. RESULTS: Under visible light protection, the solutions were stable for 14 days at room temperature whereas a loss of 20%-60% was measured after 1 day of light exposure. The loss of sennosides due to degradation can be as fast as up to 2%-2.5% per hour, which might have a tremendous impact on phytochemical analysis results during the course of an analysis. The formation of aglycones was not observed in the degradation of sennosides and rhein-8-O-glucoside. CONCLUSION: Aglycones could not be found as a result of the forced degradation. The solutions of sennosides clearly need to be protected from light to obtain reliable analytical results, and light protection is a major point for the stability of liquid preparations.

Response-Category Conflict and Control Mode Determine Memory Performance for Distractors in a Flanker Paradigm.

Cognitive conflicts can lead to better memory for task-relevant information. However, little is known about memory performance for task-irrelevant information. The present study investigated memory performance of task-irrelevant distractors using a Flanker paradigm. In two experiments (N = 699), participants first completed a study phase with 56 flanker trials. The stimuli consisted of trial-unique pictures. A congruent trial consisted of a target flanked by two identical pictures from the same category (e.g., all mammals). An incongruent trial consisted of a target and flankers from different stimulus categories (e.g., a mammal flanked by two identical birds), which results in a response-category conflict. To explore the impact of different control modes, congruent and incongruent trials were presented in pure blocks (allowing a proactive control mode) or in mixed blocks (requiring a reactive control mode). Afterwards, recognition memory was tested in a surprise memory test. In general, the results showed better memory for congruent than incongruent flankers in pure blocks. In contrast in mixed blocks, the results showed better memory for incongruent than for congruent flankers. Thus, memory performance for distractors varies systematically with response-category conflict and control mode.

Tony W. Keller (1937-2023).

Tony Keller, a pioneer in the field of Nuclear Magnetic Resonance (NMR) spectroscopy, passed away on October 27, 2023, at the age of 86 in Spiez, Switzerland. His work and vision were essential to the development and commercialization of NMR spectrometers for many areas of scientific research.

High and fast: NMR protein-proton side-chain assignments at 160 kHz and 1.2 GHz.

The NMR spectra of side-chain protons in proteins provide important information, not only about their structure and dynamics, but also about the mechanisms that regulate interactions between macromolecules. However, in the solid-state, these resonances are particularly difficult to resolve, even in relatively small proteins. We show that magic-angle-spinning (MAS) frequencies of 160 kHz, combined with a high magnetic field of 1200 MHz proton Larmor frequency, significantly improve their spectral resolution. We investigate in detail the gain for MAS frequencies between 110 and 160 kHz MAS for a model sample as well as for the hepatitis B viral capsid assembled from 120 core-protein (Cp) dimers. For both systems, we found a significantly improved spectral resolution of the side-chain region in the 1H-13C 2D spectra. The combination of 160 kHz MAS frequency with a magnetic field of 1200 MHz, allowed us to assign 61% of the aliphatic protons of Cp. The side-chain proton assignment opens up new possibilities for structural studies and further characterization of protein-protein or protein-nucleic acid interactions.

Hypnagogic states are quite common: Self-reported prevalence, modalities, and gender differences.

The hypnagogic state refers to the transitional phase between wakefulness and sleep during which vivid experiences occur. In this questionnaire study, we assessed the self-reported prevalence of hypnagogic states considering the frequency of experiences in different modalities. We also assessed the emotional quality and the vividness of the experiences. Moreover, we compared hypnagogic states to other phenomena, such as dreams, sleep paralysis, imagination, and extra-sensory perception in these measures. Hypnagogic states were reported by 80.2 % of 4456 participants and were more prevalent in women than men. Experiences were most often kinaesthetic and visual, and less often auditory, tactile, and olfactory or gustatory. Hypnagogic states were less prevalent than dreams and characterized by different modality profiles. However, they were similar to dreams in their emotional quality, the irritation they caused, and in their vividness. In conclusion, hypnagogic states are quite common.

Overshooting cognitive control adjustments in older age: Evidence from conflict- and error-related slowing in the Stroop, Simon, and flanker tasks.

Although humans gain considerable knowledge from young to older adulthood, aging is also associated with cognitive deficits. This study investigated age-related changes in dynamic cognitive control adjustments after cognitive conflicts and errors. Specifically, we compared younger and older adults' time courses of two established phenomena - post-conflict slowing and post-error slowing. Both age groups completed modified versions of three widely used cognitive conflict tasks (Stroop, Simon, and flanker task). In these tasks, occasional incongruent information triggered a conflict that had to be resolved accordingly but sometimes elicited errors. We tracked conflict- and error-related slowing across four trials after a correct conflict trial (i.e., post-conflict slowing) and an incorrect conflict trial (i.e., post-error slowing). Post-error slowing was generally stronger than post-conflict slowing. Older adults showed a disproportionally strong slowing on the first post-error trial compared to younger adults. In contrast, on subsequent trials, older adults showed a relatively stronger speed up. This pattern of results was consistent across all three tasks. The greater cross-trial response time changes in older adults suggests a deficit in fine-tuning cognitive control adjustments.

Structural conservation of HBV-like capsid proteins over hundreds of millions of years despite the shift from non-enveloped to enveloped life-style.

The discovery of nackednaviruses provided new insight into the evolutionary history of the hepatitis B virus (HBV): The common ancestor of HBV and nackednaviruses was non-enveloped and while HBV acquired an envelope during evolution, nackednaviruses remained non-enveloped. We report the capsid structure of the African cichlid nackednavirus (ACNDV), determined by cryo-EM at 3.7 Å resolution. This enables direct comparison with the known capsid structures of HBV and duck HBV, prototypic representatives of the mammalian and avian lineages of the enveloped Hepadnaviridae, respectively. The sequence identity with HBV is 24% and both the ACNDV capsid protein fold and the capsid architecture are very similar to those of the Hepadnaviridae and HBV in particular. Acquisition of the hepadnaviral envelope was thus not accompanied by a major change in capsid structure. Dynamic residues at the spike tip are tentatively assigned by solid-state NMR, while the C-terminal domain is invisible due to dynamics. Solid-state NMR characterization of the capsid structure reveals few conformational differences between the quasi-equivalent subunits of the ACNDV capsid and an overall higher capsid structural disorder compared to HBV. Despite these differences, the capsids of ACNDV and HBV are structurally highly similar despite the 400 million years since their separation.

Molecular elucidation of drug-induced abnormal assemblies of the hepatitis B virus capsid protein by solid-state NMR.

Hepatitis B virus (HBV) capsid assembly modulators (CAMs) represent a recent class of anti-HBV antivirals. CAMs disturb proper nucleocapsid assembly, by inducing formation of either aberrant assemblies (CAM-A) or of apparently normal but genome-less empty capsids (CAM-E). Classical structural approaches have revealed the CAM binding sites on the capsid protein (Cp), but conformational information on the CAM-induced off-path aberrant assemblies is lacking. Here we show that solid-state NMR can provide such information, including for wild-type full-length Cp183, and we find that in these assemblies, the asymmetric unit comprises a single Cp molecule rather than the four quasi-equivalent conformers typical for the icosahedral T = 4 symmetry of the normal HBV capsids. Furthermore, while in contrast to truncated Cp149, full-length Cp183 assemblies appear, on the mesoscopic level, unaffected by CAM-A, NMR reveals that on the molecular level, Cp183 assemblies are equally aberrant. Finally, we use a eukaryotic cell-free system to reveal how CAMs modulate capsid-RNA interactions and capsid phosphorylation. Our results establish a structural view on assembly modulation of the HBV capsid, and they provide a rationale for recently observed differences between in-cell versus in vitro capsid assembly modulation.

Probing a Hydrogen-π Interaction Involving a Trapped Water Molecule in the Solid State.

The detection and characterization of trapped water molecules in chemical entities and biomacromolecules remains a challenging task for solid materials. We herein present proton-detected solid-state Nuclear Magnetic Resonance (NMR) experiments at 100 kHz magic-angle spinning and at high static magnetic-field strengths (28.2 T) enabling the detection of a single water molecule fixed in the calix[4]arene cavity of a lanthanide complex by a combination of three types of non-covalent interactions. The water proton resonances are detected at a chemical-shift value close to zero ppm, which we further confirm by quantum-chemical calculations. Density Functional Theory calculations pinpoint to the sensitivity of the proton chemical-shift value for hydrogen-π interactions. Our study highlights how proton-detected solid-state NMR is turning into the method-of-choice in probing weak non-covalent interactions driving a whole branch of molecular-recognition events in chemistry and biology.

The left prefrontal cortex determines relevance at encoding and governs episodic memory formation.

The role hemispheric lateralization in the prefrontal cortex plays for episodic memory formation in general, and for emotionally valenced information in particular, is debated. In a randomized, double-blind, and sham-controlled design, healthy young participants (n = 254) performed 2 runs of encoding to categorize the perceptual, semantic, or emotionally valenced (positive or negative) features of words followed by a free recall and a recognition task. To resolve competing hypotheses about the contribution of each hemisphere, we modulated left or right dorsolateral prefrontal cortex (DLPFC) activity using transcranial direct current stimulation during encoding (1 mA, 20 min). With stimulation of the left DLPFC, but not the right DLPFC, encoding and free recall performance improved particularly for words that were processed semantically. In addition, enhancing left DLPFC activity increased memory formation for positive content while reducing that for negative content. In contrast, promoting right DLPFC activity increased memory formation for negative content. The left DLPFC assesses semantic properties of new memory content at encoding and thus influences how successful new episodic memories are established. Hemispheric laterlization-more active left DLPFC and less active right DLPFC-at the encoding stage shifts the formation of memory traces in favor of positively valenced content.

After-effects of responding to activated and deactivated prospective memory target events differ depending on processing overlaps.

Responding to a prospective memory task in the course of an ongoing activity requires switching tasks, which typically comes at a cost in performing the ongoing activity. Similarly, when the prospective memory task is deactivated, a cost can occur when previously relevant prospective memory targets appear in the course of the ongoing activity. In three experiments with undergraduate student participants (N = 226), in which cue focality was manipulated as a function of processing overlaps, we investigated the after-effects of activated and deactivated prospective memory target events. We predicted that lower focality results in stronger after-effects when the prospective memory task is activated but in weaker after-effects when the prospective memory task is deactivated. In contrast, we predicted that higher focality results in weaker after-effects when the prospective memory task is activated but in stronger after-effects when the prospective memory task is deactivated. For activated prospective memory, the pattern of results conformed to the expectations. For deactivated prospective memory, after-effects occurred only under high process overlap situations in a zero-target condition, in which participants were instructed for the prospective memory task, but never had the opportunity to perform it, indicating the special representational status of uncompleted intentions. We discuss these findings within the process overlap framework, which allows more fine-grained distinctions than the focal versus nonfocal dichotomy. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Cognitive load at encoding hurts memory selectivity.

People remember more task-relevant information than task-irrelevant information, and this difference can be conceptualised as memory selectivity. Selectively attending and remembering relevant information is a key ability for goal-directed behaviour and is thus critical for leading an autonomous life. In the present study, we tested the influence of cognitive load on memory selectivity. Specifically, we investigated the effects of task switching, stimulus presentation duration, and preparation time during incidental learning in five experiments (N = 351). For the study phase, we used two established task switching paradigms (cued and alternating runs). Participants were presented with picture-word pairs on which they performed one of two classification tasks. Depending on the task, participants had to attend to the picture or to the word. In a subsequent surprise recognition test, we assessed how well they remembered the targets and distractors. After 1 day or 1 week, a second recognition test assessed the longevity of the effects. Results showed that task switches (vs task repetitions), short (vs until response) stimulus duration, and short (vs long) preparation time reduced memory selectivity. The effect of preparation time was significant only in cued task switching but not in the alternating runs paradigm, highlighting the importance of advance cues for preparation effects on memory. With longer retention intervals, the effects washed out. In conclusion, higher cognitive load leads to lower selective attention and, consequently, to lower memory selectivity. The present study provides links between theories of attention, cognitive control, and memory.

The hypnagogic state: A brief update.

The hypnagogic state refers to a transitional stage between wakefulness and sleep, in which sensory perceptions can be experienced. In this review, we compile and discuss the recent scientific literature on hypnagogia research regarding the future directions proposed by Schacter (1976; Psychological Bulletin, 83, 452). After a short introduction discussing the terminology used in hypnagogia research and the differentiation of hypnagogic states with other related phenomena, we review the reported prevalence of hypnagogic states. Then, we evaluate the six future directions suggested by Schacter and we propose three further future directions. First, a better understanding of the emotional quality of hypnagogic states is needed. Second, a better understanding of why hypnagogic states occur so frequently in the visual and kinaesthetic modalities is needed. Lastly, a better understanding of the purpose of hypnagogic states is needed. In conclusion, research has made great progress in recent years, and we are one step closer to demystifying the hypnagogic state.

Fostering cognitive performance in older adults with a process- and a strategy-based cognitive training.

       The present study investigates the impact of process-based and strategy-based cognitive training to boost performance in healthy older adults. Three groups trained with either a dichotic listening training (process-based training, n = 25), an implementation intention strategy training (strategy-based training, n = 23), or served as a non-contact control group (n = 30). Our results demonstrated that training participants improved their performance in the trained tasks (process-based training: d = 3.01, strategy-based training: d = 2.6). For untrained tasks, the process-based training group showed significant working memory (d = .58) as well as episodic memory task improvement (d = 1.19) compared to the strategy-based training and to the non-contact control group (all d < .03). In contrast, in the strategy-based training group there was a tendency towards some performance gain in a fluid intelligence test (d = .92). These results indicate that cognitive training can be tailored to improve specific cognitive abilities.

Solid-State NMR Structure of Amyloid-ß Fibrils.

Amyloid fibrils are involved in a number of diseases and notably play a role in neurodegeneration, where they are present in plaques in the brain. Their structure determination might help in finding ways to interfere with their formation, and ultimately prevent disease, by revealing the structure-function relationship and helping to design molecules targeting initial assembly steps and further propagation. Here, we describe the different steps in NMR protocols which allowed the 3D structure determination of amyloid-ß fibrils.

α-Synuclein Fibril, Ribbon and Fibril-91 Amyloid Polymorphs Generation for Structural Studies.

The human α-synuclein protein, identified as one of the main markers of Parkinson's disease, is a 140-amino acid thermostable protein that can easily be overexpressed in E. coli. The purification protocol determines the ability of the protein to assemble into amyloid fibrils of well-defined structures. Here, we describe the purification and assembly protocols to obtain three well-characterized amyloid forms (ribbon, fibrils, and fibril-91) used to assess their activity in biochemical and cellular assays or to investigate their atomic structure by cryo-electron microscopy and solid-state NMR.

Attentional attenuation (rather than attentional boost) through task switching leads to a selective long-term memory decline.

Allocating attention determines what we remember later. Attentional demands vary in a task-switching paradigm, with greater demands for switch than for repeat trials. This also results in lower subsequent memory performance for switch compared to repeat trials. The main goal of the present study was to investigate the consequences of task switching after a long study-test interval and to examine the contributions of the two memory components, recollection and familiarity. In the study phase, the participants performed a task-switching procedure in which they had to switch between two classifications tasks with pictures. After a short vs. a long study-test interval of a week, the participants performed a surprise memory test for the pictures and gave remember/know judgements. The results showed that recognition memory declined after 1 week and this was mainly due to a decrease in "remember" responses. The results also showed that the task-switching effect on memory was enduring. Whereas the results of the immediate test were mixed, the results of the delayed tests showed that the task-switching effect was based on recollection, expressed in more "remember" responses for repeat than for switch trials. As recollection is more sensitive to attention manipulations than familiarity, the results align with the notion that attentional requirements at study determine what we remember, in particular after a long study-test interval.

Pharmacomodulation of a ligand targeting the HBV capsid hydrophobic pocket.

Hepatitis B virus (HBV) is a small enveloped retrotranscribing DNA virus and an important human pathogen. Its capsid-forming core protein (Cp) features a hydrophobic pocket proposed to be central notably in capsid envelopment. Indeed, mutations in and around this pocket can profoundly modulate, and even abolish, secretion of enveloped virions. We have recently shown that Triton X-100, a detergent used during Cp purification, binds to the hydrophobic pocket with micromolar affinity. We here performed pharmacomodulation of pocket binders through systematic modifications of the three distinct chemical moieties composing the Triton X-100 molecule. Using NMR and ITC, we found that the flat aromatic moiety is essential for binding, while the number of atoms of the aliphatic chain modulates binding affinity. The hydrophilic tail, in contrast, is highly tolerant to changes in both length and type. Our data provide essential information for designing a new class of HBV antivirals targeting capsid-envelope interactions.

Solid-State NMR Reveals Asymmetric ATP Hydrolysis in the Multidrug ABC Transporter BmrA.

The detailed mechanism of ATP hydrolysis in ATP-binding cassette (ABC) transporters is still not fully understood. Here, we employed 31P solid-state NMR to probe the conformational changes and dynamics during the catalytic cycle by locking the multidrug ABC transporter BmrA in prehydrolytic, transition, and posthydrolytic states, using a combination of mutants and ATP analogues. The 31P spectra reveal that ATP binds strongly in the prehydrolytic state to both ATP-binding sites as inferred from the analysis of the nonhydrolytic E504A mutant. In the transition state of wild-type BmrA, the symmetry of the dimer is broken and only a single site is tightly bound to ADP:Mg2+:vanadate, while the second site is more 'open' allowing exchange with the nucleotides in the solvent. In the posthydrolytic state, weak binding, as characterized by chemical exchange with free ADP and by asymmetric 31P-31P two-dimensional (2D) correlation spectra, is observed for both sites. Revisiting the 13C spectra in light of these findings confirms the conformational nonequivalence of the two nucleotide-binding sites in the transition state. Our results show that following ATP binding, the symmetry of the ATP-binding sites of BmrA is lost in the ATP-hydrolysis step, but is then recovered in the posthydrolytic ADP-bound state.