Visual feedback of the tongue influences speech adaptation to a physical modification of the oral cavity.

Studies examining sensorimotor adaptation of speech to changing sensory conditions have demonstrated a central role for both auditory and somatosensory feedback in speech motor learning. The potential influence of visual feedback of oral articulators, which is not typically available during speech production but may nonetheless enhance oral motor control, remains poorly understood. The present study explores the influence of ultrasound visual feedback of the tongue on adaptation of speech production (focusing on the sound /s/) to a physical perturbation of the oral articulators (prosthesis altering the shape of the hard palate). Two visual feedback groups were tested that differed in the two-dimensional plane being imaged (coronal or sagittal) during practice producing /s/ words, along with a no-visual-feedback control group. Participants in the coronal condition were found to adapt their speech production across a broader range of acoustic spectral moments and syllable contexts than the no-feedback controls. In contrast, the sagittal group showed reduced adaptation compared to no-feedback controls. The results indicate that real-time visual feedback of the tongue is spontaneously integrated during speech motor adaptation, with effects that can enhance or interfere with oral motor learning depending on compatibility of the visual articulatory information with requirements of the speaking task.

What anticipatory coarticulation in children tells us about speech motor control maturity.

PURPOSE: This study aimed to evaluate the role of motor control immaturity in the speech production characteristics of 4-year-old children, compared to adults. Specifically, two indices were examined: trial-to-trial variability, which is assumed to be linked to motor control accuracy, and anticipatory extra-syllabic vowel-to-vowel coarticulation, which is assumed to be linked to the comprehensiveness, maturity and efficiency of sensorimotor representations in the central nervous system. METHOD: Acoustic and articulatory (ultrasound) data were recorded for 20 children and 10 adults, all native speakers of Canadian French, during the production of isolated vowels and vowel-consonant-vowel (V1-C-V2) sequences. Trial-to-trial variability was measured in isolated vowels. Extra-syllabic anticipatory coarticulation was assessed in V1-C-V2 sequences by measuring the patterns of variability of V1 associated with variations in V2. Acoustic data were reported for all subjects and articulatory data, for a subset of 6 children and 2 adults. RESULTS: Trial-to-trial variability was significantly larger in children. Systematic and significant anticipation of V2 in V1 was always found in adults, but was rare in children. Significant anticipation was observed in children only when V1 was /a/, and only along the antero-posterior dimension, with a much smaller magnitude than in adults. A closer analysis of individual speakers revealed that some children showed adult-like anticipation along this dimension, whereas the majority did not. CONCLUSION: The larger trial-to-trial variability and the lack of anticipatory behavior in most children-two phenomena that have been observed in several non-speech motor tasks-support the hypothesis that motor control immaturity may explain a large part of the differences observed between speech production in adults and 4-year-old children, apart from other causes that may be linked with language development.

Sensorimotor adaptation across the speech production workspace in response to a palatal perturbation.

Talkers have been shown to adapt the production of multiple vowel sounds simultaneously in response to altered auditory feedback. The present study extends this work by exploring the adaptation of speech production to a physical alteration of the vocal tract involving a palatal prosthesis that impacts both somatosensory and auditory feedback during the production of a range of consonants and vowels. Acoustic and kinematic measures of the tongue were used to examine the impact of the physical perturbation across the various speech sounds, and to assess learned changes following 20 min of speech practice involving the production of complex, variable sentences. As in prior studies, acoustic analyses showed perturbation and adaptation effects primarily for sounds directly involving interaction with the palate. Analyses of tongue kinematics, however, revealed systematic, robust effects of the perturbation and subsequent motor learning across the full range of speech sounds. The results indicate that speakers are able to reconfigure oral motor patterns during the production of multiple speech sounds spanning the articulatory workspace following a physical alteration of the vocal tract.

Investigating how children produce rotation and pointing movements when they learn to write letters.

How do children learn to write letters? During writing acquisition, some letters may be more difficult to produce than others because certain movement sequences require more precise motor control (e.g., the rotation that produces curved lines like in letter O or the pointing movement to trace the horizontal bar of a T). Children of ages 6-10 (N = 108) wrote sequences of upper-case letters on a digitizer. They varied in the number of pointing and rotation movements. The data revealed that these movements required compensatory strategies in specific kinematic variables. For pointing movements there was a duration decrease that was compensated by an increase in in-air movement time. Rotation movements were produced with low maximal velocity but high minimal velocity. At all ages there was a global tendency to keep stability in the tempo of writing: pointing movements exhibited a duration trade-off whereas rotation movements required a trade-off on maximal and minimal velocity. The acquisition of letter writing took place between ages 6 and 7. At age 8 the children shifted focus to improving movement control. Writing automation was achieved around age 10 when the children controlled movement duration and fluency. This led to a significant increase in writing speed.

Which way to the dawn of speech?: Reanalyzing half a century of debates and data in light of speech science.

Recent articles on primate articulatory abilities are revolutionary regarding speech emergence, a crucial aspect of language evolution, by revealing a human-like system of proto-vowels in nonhuman primates and implicitly throughout our hominid ancestry. This article presents both a schematic history and the state of the art in primate vocalization research and its importance for speech emergence. Recent speech research advances allow more incisive comparison of phylogeny and ontogeny and also an illuminating reinterpretation of vintage primate vocalization data. This review produces three major findings. First, even among primates, laryngeal descent is not uniquely human. Second, laryngeal descent is not required to produce contrasting formant patterns in vocalizations. Third, living nonhuman primates produce vocalizations with contrasting formant patterns. Thus, evidence now overwhelmingly refutes the long-standing laryngeal descent theory, which pushes back "the dawn of speech" beyond ~200 ka ago to over ~20 Ma ago, a difference of two orders of magnitude.

Gene transfer from bacteria and archaea facilitated evolution of an extremophilic eukaryote.

Some microbial eukaryotes, such as the extremophilic red alga Galdieria sulphuraria, live in hot, toxic metal-rich, acidic environments. To elucidate the underlying molecular mechanisms of adaptation, we sequenced the 13.7-megabase genome of G. sulphuraria. This alga shows an enormous metabolic flexibility, growing either photoautotrophically or heterotrophically on more than 50 carbon sources. Environmental adaptation seems to have been facilitated by horizontal gene transfer from various bacteria and archaea, often followed by gene family expansion. At least 5% of protein-coding genes of G. sulphuraria were probably acquired horizontally. These proteins are involved in ecologically important processes ranging from heavy-metal detoxification to glycerol uptake and metabolism. Thus, our findings show that a pan-domain gene pool has facilitated environmental adaptation in this unicellular eukaryote.

Metabolism and metabolomics of eukaryotes living under extreme conditions.

Treatises on extremophiles are frequently focused on organisms belonging to the Archaea and Eubacteria kingdoms. However, a significant number of eukaryotes, both unicellular and multicellular, have evolved to live and thrive in extreme environments. Although less is known about eukaryotic life in extreme environments in comparison to prokaryotic extremophiles, advances in genomics and in comprehensive, high-throughput metabolic profiling techniques have provided new insight into the metabolic adaptations of eukaryotes living under extreme conditions. In this review, we will provide an overview of extremophilic life forms with emphasis on eukaryotes and we will compare metabolic adaptations in different eukaryotic extremophiles to identify generalities and specializations in adaptation to life under extreme conditions. Special emphasis will be devoted to the thermoacidophilic unicellular red alga Galdieria sulphuraria (Cyanidiaceae) as one example of a eukaryotic extremophile.

Viscosity effects on eukaryotic nitrate reductase activity.

Rate-limiting processes of catalysis by eukaryotic molybdenum-containing nitrate reductase (NaR, EC 1.7.1.1-3) were investigated using two viscosogens (glycerol and sucrose) and observing their impact on NAD(P)H:NaR activity of corn leaf NaR and recombinant Arabidopsis and yeast NaR. Holo-NaR has two "hinge" sequences between stably folded regions housing its internal electron carriers: 1) Hinge 1 between the molybdenum-containing nitrate reducing module and cytochrome b domain containing heme and 2) Hinge 2 between cytochrome b and cytochrome b reductase (CbR) module containing FAD. Solution viscosity negatively impacted the activity of these holo-NaR forms, which suggests that the rate-limiting events in catalysis were likely to involve large conformational changes that restrict or "gate" internal electron-proton transfers (IET). Little effect of viscosity was observed on recombinant CbR module and methyl viologen nitrate reduction by holo-NaR, suggesting that these activities involved no large conformational changes. To determine whether Hinge 2 is involved in gating the first step in IET, the effects of viscosogen on cytochrome c and ferricyanide reductase activities of holo-NaR and ferricyanide reductase activity of the recombinant molybdenum reductase module (CbR, Hinge 2, and cytochrome b) were analyzed. Solution viscosity negatively impacted these partial activities, as if Hinge 2 were involved in gating IET in both enzyme forms. We concluded that both Hinges 1 and 2 appear to be involved in gating IET steps by restricting the movement of the cytochrome b domain relative to the larger nitrate-reducing and electron-donating modules of NaR.

Structural basis of eukaryotic nitrate reduction: crystal structures of the nitrate reductase active site.

Nitrate assimilation in autotrophs provides most of the reduced nitrogen on earth. In eukaryotes, reduction of nitrate to nitrite is catalyzed by the molybdenum-containing NAD(P)H:nitrate reductase (NR; EC 1.7.1.1-3). In addition to the molybdenum center, NR contains iron-heme and flavin adenine dinucleotide as redox cofactors involved in an internal electron transport chain from NAD(P)H to nitrate. Recombinant, catalytically active Pichia angusta nitrate-reducing, molybdenum-containing fragment (NR-Mo) was expressed in P. pastoris and purified. Crystal structures for NR-Mo were determined at 1.7 and 2.6 angstroms. These structures revealed a unique slot for binding nitrate in the active site and identified key Arg and Trp residues potentially involved in nitrate binding. Dimeric NR-Mo is similar in overall structure to sulfite oxidases, with significant differences in the active site. Sulfate bound in the active site caused conformational changes, as compared with the unbound enzyme. Four ordered water molecules located in close proximity to Mo define a nitrate binding site, a penta-coordinated reaction intermediate, and product release. Because yeast NAD(P)H:NR is representative of the family of eukaryotic NR, we propose a general mechanism for nitrate reduction catalysis.

Comparative genomics of two closely related unicellular thermo-acidophilic red algae, Galdieria sulphuraria and Cyanidioschyzon merolae, reveals the molecular basis of the metabolic flexibility of Galdieria sulphuraria and significant differences in carbohydrate metabolism of both algae.

Unicellular algae serve as models for the study and discovery of metabolic pathways, for the functional dissection of cell biological processes such as organellar division and cell motility, and for the identification of novel genes and gene functions. The recent completion of several algal genome sequences and expressed sequence tag collections and the establishment of nuclear and organellar transformation methods has opened the way for functional genomics approaches using algal model systems. The thermo-acidophilic unicellular red alga Galdieria sulphuraria represents a particularly interesting species for a genomics approach owing to its extraordinary metabolic versatility such as heterotrophic and mixotrophic growth on more than 50 different carbon sources and its adaptation to hot acidic environments. However, the ab initio prediction of genes required for unknown metabolic pathways from genome sequences is not trivial. A compelling strategy for gene identification is the comparison of similarly sized genomes of related organisms with different physiologies. Using this approach, candidate genes were identified that are critical to the metabolic versatility of Galdieria. Expressed sequence tags and high-throughput genomic sequence reads covering >70% of the G. sulphuraria genome were compared to the genome of the unicellular, obligate photoautotrophic red alga Cyanidioschyzon merolae. More than 30% of the Galdieria sequences did not relate to any of the Cyanidioschyzon genes. A closer inspection of these sequences revealed a large number of membrane transporters and enzymes of carbohydrate metabolism that are unique to Galdieria. Based on these data, it is proposed that genes involved in the uptake of reduced carbon compounds and enzymes involved in their metabolism are crucial to the metabolic flexibility of G. sulphuraria.

Purification and biochemical characterization of simplified eukaryotic nitrate reductase expressed in Pichia pastoris.

NAD(P)H:nitrate reductase (NaR, EC 1.7.1.1-3) is a useful enzyme in biotechnological applications, but it is very complex in structure and contains three cofactors-flavin adenine dinucleotide, heme-Fe, and molybdenum-molybdopterin (Mo-MPT). A simplified nitrate reductase (S-NaR1) consisting of Mo-MPT-binding site and nitrate-reducing active site was engineered from yeast Pichia angusta NaR cDNA (YNaR1). S-NaR1 was cytosolically expressed in high-density fermenter culture of methylotrophic yeast Pichia pastoris. Total amount of S-NaR1 protein produced was approximately 0.5 g per 10 L fermenter run, and methanol phase productivity was 5 microg protein/g wet cell weight/h. Gene copy number in genomic DNA of different clones showed direct correlation with the expression level. S-NaR1 was purified to homogeneity in one step by immobilized metal affinity chromatography (IMAC) and total amount of purified protein per run of fermentation was approximately 180 mg. Polypeptide size was approximately 55 kDa from electrophoretic analysis, and S-NaR1 was mainly homo-tetrameric in its active form, as shown by gel filtration. S-NaR1 accepted electrons efficiently from reduced bromphenol blue (kcat = 2081 s(-1)) and less so from reduced methyl viologen (kcat = 159 s(-1)). The nitrate KM for S-NaR1 was 30 +/- 3 microM, which is very similar to YNaR1. S-NaR1 is capable of specific nitrate reduction, and direct electric current, as shown by catalytic nitrate reduction using protein film cyclic voltammetry, can drive this reaction. Thus, S-NaR1 is an ideal form of this enzyme for commercial applications, such as an enzymatic nitrate biosensor formulated with S-NaR1 interfaced to an electrode system.