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Identification and Characterization of Three Epithiospecifier Protein Isoforms in Brassica oleracea
(2019)
Glucosinolates present in Brassicaceae play a major role in herbivory defense. Upon tissue disruption, glucosinolates come into contact with myrosinase, which initiates their breakdown to biologically active compounds. Among these, the formation of epithionitriles is triggered by the presence of epithiospecifier protein (ESP) and a terminal double bond in the glucosinolate side chain. One ESP gene is characterized in the model plant Arabidopsis thaliana (AtESP; At1g54040.2). However, Brassica species underwent genome triplication since their divergence from the Arabidopsis lineage. This indicates the presence of multiple ESP isoforms in Brassica crops that are currently poorly characterized. We identified three B. oleracea ESPs, specifically BoESP1 (LOC106296341), BoESP2 (LOC106306810), and BoESP3 (LOC106325105) based on in silico genome analysis. Transcript and protein abundance were assessed in shoots and roots of four B. oleracea vegetables, namely broccoli, kohlrabi, white, and red cabbage, because these genotypes showed a differential pattern for the formation of glucosinolate hydrolysis products as well for their ESP activity. BoESP1 and BoESP2 were expressed mainly in shoots, while BoESP3 was abundant in roots. Biochemical characterization of heterologous expressed BoESP isoforms revealed different substrate specificities towards seven glucosinolates: all isoforms showed epithiospecifier activity on alkenyl glucosinolates, but not on non-alkenyl glucosinolates. The pH-value differently affected BoESP activity: while BoESP1 and BoESP2 activities were optimal at pH 6-7, BoESP3 activity remained relatively stable from pH 4 to 7. In order test their potential for the in vivo modification of glucosinolate breakdown, the three isoforms were expressed in A. thaliana Hi-0, which lacks AtESP expression, and analyzed for the effect on their respective hydrolysis products. The BoESPs altered the hydrolysis of allyl glucosinolate in the A. thaliana transformants to release 1-cyano-2,3-epithiopropane and reduced formation of the corresponding 3-butenenitrile and allyl isothiocyanate. Plants expressing BoESP2 showed the highest percentage of released epithionitriles. Given these results, we propose a model for isoform-specific roles of B. oleracea ESPs in glucosinolate breakdown.
Compulsive behaviors (e.g., addiction) can be viewed as an aberrant decision process where inflexible reactions automatically evoked by stimuli (habit) take control over decision making to the detriment of a more flexible (goal-oriented) behavioral learning system. These behaviors are thought to arise from learning algorithms known as "model-based" and "model-free" reinforcement learning. Gambling disorder, a form of addiction without the confound of neurotoxic effects of drugs, showed impaired goal-directed control but the way in which problem gamblers (PG) orchestrate model-based and model-free strategies has not been evaluated. Forty-nine PG and 33 healthy participants (CP) completed a two-step sequential choice task for which model-based and model-free learning have distinct and identifiable trial-by-trial learning signatures. The influence of common psychopathological comorbidities on those two forms of learning were investigated. PG showed impaired model-based learning, particularly after unrewarded outcomes. In addition, PG exhibited faster reaction times than CP following unrewarded decisions. Troubled mood, higher impulsivity (i.e., positive and negative urgency) and current and chronic stress reported via questionnaires did not account for those results. These findings demonstrate specific reinforcement learning and decision-making deficits in behavioral addiction that advances our understanding and may be important dimensions for designing effective interventions.
Protein interaction and protein imaging strongly benefit from the advancements in time-resolved and superresolution fluorescence microscopic techniques. However, the techniques were typically applied separately and ex vivo because of technical challenges and the absence of suitable fluorescent protein pairs. Here, we show correlative in vivo fluorescence lifetime imaging microscopy Forster resonance energy transfer (FLIM-FRET) and stimulated emission depletion (STED) microscopy to unravel protein mechanics and structure in living cells. We use magnetotactic bacteria as a model system where two proteins, MamJ and MamK, are used to assemble magnetic particles called magnetosomes. The filament polymerizes out of MamK and the magnetosomes are connected via the linker MamJ. Our system reveals that bacterial filamentous structures are more fragile than the connection of biomineralized particles to this filament. More importantly, we anticipate the technique to find wide applicability for the study and quantification of biological processes in living cells and at high resolution.
In an overt visual priming experiment, we investigate the role of orthography in native (L1) and non-native (L2) processing of German morphologically complex words. We compare priming effects for inflected and derived morphologically related prime-target pairs versus otherwise matched, purely orthographically related pairs. The results show morphological priming effects in both the L1 and L2 group, with no significant difference between inflection and derivation. However, L2 speakers, but not L1 speakers, also showed significant priming for orthographically related pairs. Our results support the claim that L2 speakers focus more on surface-level information such as orthography during visual word recognition. This can cause orthographic priming effects in morphologically related prime-target pairs, which may conceal L1-L2 differences in morphological processing.
This paper challenges the solely rational view of the scenario technique as a strategy and foresight tool designed to cope with uncertainty by considering multiple possible future states. The paper employs an affordance-based view that allows for the identification and structuring of hidden, emergent attributes of the scenario technique beyond the intended ones. The suggested framework distinguishes between affordances (1) that are intended by the organization and relate to its goals, (2) that emergently generate organizational benefits, and (3) that do not relate to organizational but individual interests. Also, constraints in the use of scenarios are discussed. Affordance theory’s specific lens shows that the emergence of such attributes depends on the users’ specific intentions.
Humans generate internal models of their environment to predict events in the world. As the environments change, our brains adjust to these changes by updating their internal models. Here, we investigated whether and how 9-month-old infants differentially update their models to represent a dynamic environment. Infants observed a predictable sequence of stimuli, which were interrupted by two types of cues. Following the update cue, the pattern was altered, thus, infants were expected to update their predictions for the upcoming stimuli. Because the pattern remained the same after the no-update cue, no subsequent updating was required. Infants showed an amplified negative central (Nc) response when the predictable sequence was interrupted. Late components such as the PSW were also evoked in response to unexpected stimuli; however, we found no evidence for a differential response to the informational value of surprising cues at later stages of processing. Infants rather learned that surprising cues always signal a change in the environment that requires updating. Interestingly, infants responded with an amplified neural response to the absence of an expected change, suggesting a top-down modulation of early sensory processing in infants. Our findings corroborate emerging evidence showing that infants build predictive models early in life.