Controlling stimulus ambiguity reduces spurious creative ideation variance in a cyclic adaptation of the alternative uses task.

In the alternative uses task (AUT), a well-established creativity assessment, participants propose alternative uses for common items (e.g., a brick) within a 2-3 min timeframe. While idea evaluation is likely involved, the emphasis is strongly on idea generation. Here, we test the value of presenting a word overlapping an image compared to a word only prompt, and we introduce a cyclic adaptation of the AUT explicitly calling on participants to choose their best idea. In Experiment 1, as compared to word only, word + image prompts increased idea fluency but reduced idea originality and variability within a group of native Polish speakers. Thus, word + image prompts improve AUT baselining. In Experiment 2, different participants produced as many ideas as possible within two minutes (List) or their single best idea at the end of each of three 30 s ideation cycles (Cycle). Although originality did not differ between List and Cycle overall, the first three ideas in List were rated as less creative than the ideas in Cycle. Overall, we conclude that using disambiguating images reduces spurious interindividual variability in the AUT while introducing idea evaluation in the task allows us to assess creativity beyond idea generation.

Brain potentials reveal how emotion filters native language access when bilinguals read words in their second language.

It is now well established that reading words in a second language (L2) automatically activates native language (L1) translations in bilinguals. Although there is evidence that access to such representations is inhibited when words have a negative emotional valence, the mechanism underlying such inhibition is elusive, and it is unknown whether inhibition arises online as L2 is being processed or whether negative valence affects subsequent L1 processing. Here, we recorded event-related brain potentials in Chinese-English bilinguals engaged in an implicit translation-priming paradigm involving L2 (English) word pairs. Participants performed a semantic relatedness task, unaware that word pairs could conceal a sound repetition if translated into Chinese. When emotional valence was manipulated in prime position (first word), we observed form repetition priming through L1 translations for positive but not for negative words. However, when emotional valence was manipulated in target position (second word), priming occurred for both positive and negative word valences. This result begins to elucidate the mechanism by which emotion regulates language processing in bilinguals: Negative words in L2 induce a refractory period during which cross-language lexical access is blocked. These findings show that despite being neuroanatomically distinct in the human brain, emotional (limbic) regulation systems can penetrate language processing.

How stereotype threat affects the brain dynamics of creative thinking in female students.

When people are placed in a situation where they are at risk of substantiating a negative stereotype about their social group (a scenario termed stereotype threat), the extra pressure to avoid this outcome can undermine their performance. Substantial and consistent gender disparities in STEM fields leave women vulnerable to stereotype threat, including the stereotype that women are not as good at generating creative and innovative ideas as men. We tested whether female students' creative thinking is affected by a stereotype threat by measuring power in the alpha frequency band (8-12Hz oscillations) that has been associated with better creative thinking outcomes. Counter to expectations that a stereotype threat would reduce alpha power associated with creative thinking, analyses showed increased alpha power following the introduction of the stereotype threat. This outcome suggests that women may have attempted to increase their internal attention during the task in order to disprove the stereotype. Behaviorally, this effort did not lead to changes in creative performance, suggesting that the stereotype threat decoupled alpha power from creative thinking outcomes. These results support a growing school of thought in the neuroscience of creativity literature that the alpha power often seen in conjunction with creative behavior is not necessarily related to the creativity processes themselves, but rather might be part of a larger network modulating the distribution of attentional resources more broadly.

High-resolution association mapping with libraries of immortalized lines from ancestral landraces.

KEY MESSAGE: Association mapping with immortalized lines of landraces offers several advantages including a high mapping resolution, as demonstrated here in maize by identifying the causal variants underlying QTL for oil content and the metabolite allantoin. Landraces are traditional varieties of crops that present a valuable yet largely untapped reservoir of genetic variation to meet future challenges of agriculture. Here, we performed association mapping in a panel comprising 358 immortalized maize lines from six European Flint landraces. Linkage disequilibrium decayed much faster in the landraces than in the elite lines included for comparison, permitting a high mapping resolution. We demonstrate this by fine-mapping a quantitative trait locus (QTL) for oil content down to the phenylalanine insertion F469 in DGAT1-2 as the causal variant. For the metabolite allantoin, related to abiotic stress response, we identified promoter polymorphisms and differential expression of an allantoinase as putative cause of variation. Our results demonstrate the power of this approach to dissect QTL potentially down to the causal variants, toward the utilization of natural or engineered alleles in breeding. Moreover, we provide guidelines for studies using ancestral landraces for crop genetic research and breeding.

Glucosylation of (±)-Menthol by Uridine-Diphosphate-Sugar Dependent Glucosyltransferases from Plants.

Menthol is a cyclic monoterpene alcohol of the essential oils of plants of the genus Mentha, which is in demand by various industries due to its diverse sensorial and physiological properties. However, its poor water solubility and its toxic effect limit possible applications. Glycosylation offers a solution as the binding of a sugar residue to small molecules increases their water solubility and stability, renders aroma components odorless and modifies bioactivity. In order to identify plant enzymes that catalyze this reaction, a glycosyltransferase library containing 57 uridine diphosphate sugar-dependent enzymes (UGTs) was screened with (±)-menthol. The identity of the products was confirmed by mass spectrometry and nuclear magnetic resonance spectroscopy. Five enzymes were able to form (±)-menthyl-ß-d-glucopyranoside in whole-cell biotransformations: UGT93Y1, UGT93Y2, UGT85K11, UGT72B27 and UGT73B24. In vitro enzyme activity assays revealed highest catalytic activity for UGT93Y1 (7.6 nkat/mg) from Camellia sinensis towards menthol and its isomeric forms. Although UGT93Y2 shares 70% sequence identity with UGT93Y1, it was less efficient. Of the five enzymes, UGT93Y1 stood out because of its high in vivo and in vitro biotransformation rate. The identification of novel menthol glycosyltransferases from the tea plant opens new perspectives for the biotechnological production of menthyl glucoside.

Six Uridine-Diphosphate Glycosyltransferases Catalyze the Glycosylation of Bioactive C13-Apocarotenols.

C13-apocarotenoids (norisoprenoids) are carotenoid-derived oxidation products that perform important physiological functions in plants. Although their biosynthetic pathways have been extensively studied, their metabolism including glycosylation remains poorly understood. Candidate uridine-diphosphate glycosyltransferase genes (UGTs) were selected based on their high transcript abundance in comparison with other UGTs in vegetative tissues of Nicotiana benthamiana and peppermint (Mentha × piperita), as these tissues are rich sources of apocarotenoid glucosides. Hydroxylated C13-apocarotenol substrates were produced by P450-catalyzed biotransformation and microbial/plant enzyme systems were established for the synthesis of glycosides. Natural substrates were identified by physiological aglycone libraries prepared from isolated plant glycosides. In total, we identified six UGTs that catalyze the glucosylation of C13-apocarotenols, where Glc is bound either to the cyclohexene ring or the butane side chain. MpUGT86C10 is a superior novel enzyme that catalyzes the glucosylation of allelopathic 3-hydroxy-α-damascone, 3-oxo-α-ionol, 3-oxo-7,8-dihydro-α-ionol (Blumenol C), and 3-hydroxy-7,8-dihydro-ß-ionol, whereas a germination test demonstrated the higher phytotoxic potential of a norisoprenoid glucoside in comparison to its aglycone. Glycosylation of C13-apocarotenoids has several functions in plants, including increased allelopathic activity of the aglycone, facilitating exudation by roots and allowing symbiosis with arbuscular mycorrhizal fungi. The results enable in-depth analysis of the roles of glycosylated norisoprenoid allelochemicals, the physiological functions of apocarotenoids during arbuscular mycorrhizal colonization, and the associated maintenance of carotenoid homeostasis.

Engineering creativity: Prior experience modulates electrophysiological responses to novel metaphors.

Novel metaphorical language use exemplifies human creativity through production and comprehension of meaningful linguistic expressions that may have never been heard before. Available electrophysiological research demonstrates, however, that novel metaphor comprehension is cognitively costly, as it requires integrating information from distantly related concepts. Herein, we investigate if such cognitive cost may be reduced as a factor of prior domain knowledge. To this end, we asked engineering and nonengineering students to read for comprehension literal, novel metaphorical, and anomalous sentences related to engineering or general knowledge, while undergoing EEG recording. Upon reading each sentence, participants were asked to judge whether or not the sentence was original in meaning (novelty judgment) and whether or not it made sense (sensicality judgment). When collapsed across groups, our findings demonstrate a gradual N400 modulation with N400 being maximal in response to anomalous, followed by metaphorical, and literal sentences. Between-group comparisons revealed a mirror effect on the N400 to novel metaphorical sentences, with attenuated N400 in engineers and enhanced N400 in non-engineers. Critically, planned comparisons demonstrated reduced N400 amplitudes to engineering novel metaphors in engineers relative to non-engineers, pointing to an effect of prior knowledge on metaphor processing. This reduction, however, was observed in the absence of a sentence type × knowledge × group interaction. Altogether, our study provides novel evidence suggesting that prior domain knowledge may have a direct impact on creative language comprehension.

Keep calm and carry on: electrophysiological evaluation of emotional anticipation in the second language.

Investigations of the so-called 'foreign language effect' have shown that emotional experience is language-dependent in bilingual individuals. Response to negative experiences, in particular, appears attenuated in the second language (L2). However, the human brain is not only reactive, but it also builds on past experiences to anticipate future events. Here, we investigated affective anticipation in immersed Polish-English bilinguals using a priming paradigm in which a verbal cue of controlled affective valence allowed making predictions about a subsequent picture target. As expected, native word cues with a negative valence increased the amplitude of the stimulus preceding negativity, an electrophysiological marker of affective anticipation, as compared with neutral ones. This effect was observed in Polish-English bilinguals and English monolinguals alike. The contrast was non-significant when Polish participants were tested in English, suggesting a possible reduction in affective sensitivity in L2. However, this reduction was not validated by a critical language × valence interaction in the bilingual group, possibly because they were highly fluent in English and because the affective stimuli used in the present study were particularly mild. These results, which are neither fully consistent nor inconsistent with the foreign language effect, provide initial insights into the electrophysiology of affective anticipation in bilingualism.

Novel biotechnological glucosylation of high-impact aroma chemicals, 3(2H)- and 2(5H)-furanones.

Glucosyltransferases are versatile biocatalysts to chemically modify small molecules and thus enhance their water solubility and structural stability. Although the genomes of all organisms harbor a multitude of glucosyltransferase genes, their functional characterization is hampered by the lack of high-throughput in-vivo systems to rapidly test the versatility of the encoded proteins. We have developed and applied a high-throughput whole cell biotransformation system to screen a plant glucosyltransferase library. As proof of principle, we identified 25, 24, 15, and 18 biocatalysts transferring D-glucose to sotolone, maple furanone, furaneol and homofuraneol, four highly appreciated flavor compounds, respectively. Although these 3(2H)- and 2(5H)-furanones have extremely low odor thresholds their glucosides were odorless. Upscaling of the biotechnological process yielded titers of 5.3 and 7.2 g/L for the new to nature ß-D-glucopyranosides of sotolone and maple furanone, respectively. Consequently, plant glucosyltransferase show stunning catalytic activities, which enable the economical production of novel and unexplored chemicals with exciting new functionalities by whole-cell biotransformation.

The bilingual brain turns a blind eye to negative statements in the second language.

Neurobilingualism research has failed to reveal significant language differences in the processing of affective content. However, the evidence to date derives mostly from studies in which affective stimuli are presented out of context, which is unnatural and fails to capture the complexity of everyday sentence-based communication. Here we investigated semantic integration of affectively salient stimuli in sentential context in the first- and second-language (L2) of late fluent Polish-English bilinguals living in the UK. The 19 participants indicated whether Polish and English sentences ending with a semantically and affectively congruent or incongruent adjective of controlled affective valence made sense while undergoing behavioral and electrophysiological recordings. We focused on the N400, a wave of event-related potentials known to index semantic integration. We expected N400 amplitude to index increased processing demands in L2 English comprehension and potential language-valence interactions to reveal differences in affective processing between languages. Contrary to our initial expectation, we found increased N400 for sentences in L1 Polish, possibly driven by greater affective salience of sentences in the native language. Critically, language interacted with affective valence, such that N400 amplitudes were reduced for English sentences ending in a negative fashion as compared to all other conditions. We interpreted this as a sign that bilinguals suppress L2 content embedded in naturalistic L2 sentences when it has negative valence, thus extending the findings of previous research on single words in clinical and linguistic research.

Hemispheric asymmetry of emotion words in a non-native mind: a divided visual field study.

This study investigates hemispheric specialization for emotional words among proficient non-native speakers of English by means of the divided visual field paradigm. The motivation behind the study is to extend the monolingual hemifield research to the non-native context and see how emotion words are processed in a non-native mind. Sixty eight females participated in the study, all highly proficient in English. The stimuli comprised 12 positive nouns, 12 negative nouns, 12 non-emotional nouns and 36 pseudo-words. To examine the lateralization of emotion, stimuli were presented unilaterally in a random fashion for 180 ms in a go/no-go lexical decision task. The perceptual data showed a right hemispheric advantage for processing speed of negative words and a complementary role of the two hemispheres in the recognition accuracy of experimental stimuli. The data indicate that processing of emotion words in non-native language may require greater interhemispheric communication, but at the same time demonstrates a specific role of the right hemisphere in the processing of negative relative to positive valence. The results of the study are discussed in light of the methodological inconsistencies in the hemifield research as well as the non-native context in which the study was conducted.

OEP61 is a chaperone receptor at the plastid outer envelope.

Chloroplast precursor proteins encoded in the nucleus depend on their targeting sequences for delivery to chloroplasts. There exist different routes to the chloroplast outer envelope, but a common theme is the involvement of molecular chaperones. Hsp90 (heat-shock protein 90) delivers precursors via its receptor Toc64, which transfers precursors to the core translocase in the outer envelope. In the present paper, we identify an uncharacterized protein in Arabidopsis thaliana OEP61 which shares common features with Toc64, and potentially provides an alternative route to the chloroplasts. Sequence analysis indicates that OEP61 possesses a clamp-type TPR (tetratricopeptide repeat) domain capable of binding molecular chaperones, and a C-terminal TMD (transmembrane domain). Phylogenetic comparisons show sequence similarities between the TPR domain of OEP61 and those of the Toc64 family. Expression of mRNA and protein was detected in all plant tissues, and localization at the chloroplast outer envelope was demonstrated by a combination of microscopy and in vitro import assays. Binding assays show that OEP61 interacts specifically with Hsp70 (heat-shock protein 70) via its TPR clamp domain. Furthermore, OEP61 selectively recognizes chloroplast precursors via their targeting sequences, and a soluble form of OEP61 inhibits chloroplast targeting. We therefore propose that OEP61 is a novel chaperone receptor at the chloroplast outer envelope, mediating Hsp70-dependent protein targeting to chloroplasts.

A novel isoform of pantothenate synthetase in the Archaea.

The linear biosynthetic pathway leading from alpha-ketoisovalerate to pantothenate (vitamin B5) and on to CoA comprises eight steps in the Bacteria and Eukaryota. Genes for up to six steps of this pathway can be identified by sequence homology in individual archaeal genomes. However, there are no archaeal homologs to known isoforms of pantothenate synthetase (PS) or pantothenate kinase. Using comparative genomics, we previously identified two conserved archaeal protein families as the best candidates for the missing steps. Here we report the characterization of the predicted PS gene from Methanosarcina mazei, which encodes a hypothetical protein (MM2281) with no obvious homologs outside its own family. When expressed in Escherichia coli, MM2281 partially complemented an auxotrophic mutant without PS activity. Purified recombinant MM2281 showed no PS activity on its own, but the enzyme enabled substantial synthesis of [14C]4'-phosphopantothenate from [14C]beta-alanine, pantoate and ATP when coupled with E. coli pantothenate kinase. ADP, but not AMP, was detected as a coproduct of the coupled reaction. MM2281 also transferred the 14C-label from [14C]beta-alanine to pantothenate in the presence of pantoate and ADP, presumably through isotope exchange. No exchange took place when pantoate was removed or ADP replaced with AMP. Our results indicate that MM2281 represents a novel type of PS that forms ADP and is strongly inhibited by its product pantothenate. These properties differ substantially from those of bacterial PS, and may explain why PS genes, in contrast to other pantothenate biosynthetic genes, were not exchanged horizontally between the Bacteria and Archaea.

Elucidation of the final reactions of DIMBOA-glucoside biosynthesis in maize: characterization of Bx6 and Bx7.

Benzoxazinoids were identified in the early 1960s as secondary metabolites of the grasses that function as natural pesticides and exhibit allelopathic properties. Benzoxazinoids are synthesized in seedlings and stored as glucosides (glcs); the main aglucone moieties are 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one (DIBOA) and 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA). The genes of DIBOA-glc biosynthesis have previously been isolated and the enzymatic functions characterized. Here, the enzymes for conversion of DIBOA-glc to DIMBOA-glc are identified. DIBOA-glc is the substrate of the dioxygenase BENZOXAZINLESS6 (BX6) and the produced 2,4,7-trihydroxy-2H-1,4-benzoxazin-3-(4H)-one-glc is metabolized by the methyltransferase BX7 to yield DIMBOA-glc. Both enzymes exhibit moderate K(m) values (below 0.4 mm) and k(cat) values of 2.10 s(-1) and 0.25 s(-1), respectively. Although BX6 uses a glucosylated substrate, our localization studies indicate a cytoplasmic localization of the dioxygenase. Bx6 and Bx7 are highest expressed in seedling tissue, a feature shared with the other Bx genes. At present, Bx6 and Bx7 have no close relatives among the members of their respective gene families. Bx6 and Bx7 map to the cluster of Bx genes on the short arm of chromosome 4.

Pantothenate synthetase is essential but not limiting for pantothenate biosynthesis in Arabidopsis.

Pantothenate (vitamin B5) is the universal precursor for coenzyme A (CoA), an essential cofactor that is required in the metabolism of carbohydrates and fatty acids. The final step of bacterial and eukaryotic pantothenate biosynthesis is catalyzed by pantothenate synthetase (PTS), which is encoded by a single gene in Arabidopsis thaliana (AtPTS). There was debate whether PTS represents the only mode of pantothenate production because previous biochemical evidence pointed to an additional pantothenate pathway in plants. Here we show that insertional mutant alleles of AtPTS confer a recessive embryo-lethal phenotype with mutant embryos arrested at the preglobular stage. Exogenous pantothenate was required for normal seed development and germination and also facilitated the remaining life cycle. Complementation of the mutant phenotype was likewise achieved by heterologous expression of E. coli PTS (panC). The panC transgene increased the total PTS activity in leaves by up to 500-fold but did not affect the steady-state level of pantothenate, indicating that PTS is essential but not limiting for pantothenate production. The auxotrophic AtPTS knockout phenotype suggests that the embryo and possibly all other tissues are autonomous for the biosynthesis of pantothenate. This view is consistent with the near-ubiquitous expression of AtPTS as judged by promoter:beta-glucuronidase analysis. Given the high demand for CoA during storage oil accumulation, we analyzed transcript and metabolite patterns of CoA biosynthesis in seeds. The data indicate that the pantothenate and CoA contents follow distinct developmental programs and that both transcriptional and posttranslational control mechanisms are important for CoA homeostasis.

Molecular adaptation and allostery in plant pantothenate synthetases.

Pantothenate synthetase catalyzes the ATP-dependent condensation of pantoate and beta-alanine to yield pantothenate, the essential precursor to coenzyme A. Bacterial and plant pantothenate synthetases are dimeric enzymes that share significant sequence identity. Here we show that the two-step reaction mechanism of pantothenate synthetase is conserved between the enzymes from Arabidopsis thaliana and Escherichia coli. Strikingly, though, the Arabidopsis enzyme exhibits large allosteric effects, whereas the Escherichia coli enzyme displays essentially non-allosteric behavior. Our data suggest that specific subunit contacts were selected and maintained in the plant lineage of the pantothenate synthetase protein family and that the resulting allosteric interactions are balanced for efficient catalysis at low pantoate levels. This is supported by mutations in the putative subunit interface of Arabidopsis pantothenate synthetase, which strongly attenuated or otherwise modified its allosteric properties but did not affect the dimeric state of the enzyme. At the molecular level, plant pantothenate synthetases exemplify functional adaptation through allostery and without alterations to the active site architecture. We propose that the allosteric behavior confers a selective advantage in the context of the subcellular compartmentation of pantothenate biosynthesis in plants.