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1.  REV, A BRET-Based Sensor of ERK Activity 
Networks of signaling molecules are activated in response to environmental changes. How are these signaling networks dynamically integrated in space and time to process particular information? To tackle this issue, biosensors of single signaling pathways have been engineered. Bioluminescence resonance energy transfer (BRET)-based biosensors have proven to be particularly efficient in that matter due to the high sensitivity of this technology to monitor protein–protein interactions or conformational changes in living cells. Extracellular signal-regulated kinases (ERK) are ubiquitously expressed and involved in many diverse cellular functions that might be encoded by the strength and spatio-temporal pattern of ERK activation. We developed a BRET-based sensor of ERK activity, called Rluc8-ERKsubstrate-Venus (REV). As expected, BRET changes of REV were correlated with ERK phosphorylation, which is required for its kinase activity. In neurons, the nature of the stimuli determines the strength, the location, or the moment of ERK activation, thus highlighting how acute modulation of ERK may encode the nature of initial stimulus to specify the consequences of this activation. This study provides evidence for suitability of REV as a new biosensor to address biological questions.
PMCID: PMC3727045  PMID: 23908646
biosensor; bioluminescence resonance energy transfer; BRET imaging; fluorescence lifetime imaging microscopy; extracellular signal-regulated kinases; spatio-temporal signaling; Rluc8-ERKsubstrate-Venus
2.  Temporal Decoding of MAP Kinase and CREB Phosphorylation by Selective Immediate Early Gene Expression 
PLoS ONE  2013;8(3):e57037.
A wide range of growth factors encode information into specific temporal patterns of MAP kinase (MAPK) and CREB phosphorylation, which are further decoded by expression of immediate early gene products (IEGs) to exert biological functions. However, the IEG decoding system remain unknown. We built a data-driven based on time courses of MAPK and CREB phosphorylation and IEG expression in response to various growth factors to identify how signal is processed. We found that IEG expression uses common decoding systems regardless of growth factors and expression of each IEG differs in upstream dependency, switch-like response, and linear temporal filters. Pulsatile ERK phosphorylation was selectively decoded by expression of EGR1 rather than c-FOS. Conjunctive NGF and PACAP stimulation was selectively decoded by synergistic JUNB expression through switch-like response to c-FOS. Thus, specific temporal patterns and combinations of MAPKs and CREB phosphorylation can be decoded by selective IEG expression via distinct temporal filters and switch-like responses. The data-driven modeling is versatile for analysis of signal processing and does not require detailed prior knowledge of pathways.
PMCID: PMC3587639  PMID: 23469182
3.  What’s “right” in language comprehension: ERPs reveal right hemisphere language capabilities 
Although the term “nonverbal” is often applied to the right cerebral hemisphere (RH), a growing body of work indicates that the RH can comprehend language and, indeed, that it makes critical contributions to normal language functions. Reviewed here are studies that have examined RH language capabilities by combining visual half-field presentation methods with event-related potential (ERP) measures. Because they afford temporal and functional specificity and can be obtained as participants simply process language for meaning, ERPs provide especially valuable insights into RH language functions. Such studies suggest that the RH appreciates word and message-level meaning information, and that it may play a particularly important role in the processing of relatively unpredictable semantic relationships. In addition, this work suggests that patterns observed for everyday language processing may often be an emergent property of multiple, distinct mechanisms operating in parallel as the left and right hemispheres jointly comprehend language.
PMCID: PMC2748422  PMID: 19777128
4.  The green beards of language 
Ecology and Evolution  2013;3(4):1104-1112.
Language transfers information on at least three levels; (1) what is said, (2) how it is said (what language is used), and, (3) that it is said (that speaker and listener both possess the ability to use language). The use of language is a form of honest cooperation on two of these levels; not necessarily on what is said, which can be deceitful, but always on how it is said and that it is said. This means that the language encoding and decoding systems had to evolve simultaneously, through mutual fitness benefits. Theoretical problems surrounding the evolution of cooperation disappear if a recognition system is present enabling cooperating individuals to identify each other – if they are equipped with “green beards”. Here, I outline how both the biological and cultural aspects of language are bestowed with such recognition systems. The biological capacities required for language signal their presence through speech and understanding. This signaling cannot be invaded by “false green beards” because the traits and the signal of their presence are one and the same. However, the real usefulness of language comes from its potential to convey an infinite number of meanings through the dynamic handling of symbols – through language itself. But any specific language also signals its presence to others through usage and understanding. Thus, languages themselves cannot be invaded by “false green beards” because, again, the trait and the signal of its presence are one and the same. These twin green beards, in both the biological and cultural realms, are unique to language.
PMCID: PMC3631417  PMID: 23610647
Chloropogonology; cooperation; Hamilton's rule; language; reciprocity
5.  Rho Kinase's Role in Myosin Recruitment to the Equatorial Cortex of Mitotic Drosophila S2 Cells Is for Myosin Regulatory Light Chain Phosphorylation 
PLoS ONE  2006;1(1):e131.
Myosin II recruitment to the equatorial cortex is one of the earliest events in establishment of the cytokinetic contractile ring. In Drosophila S2 cells, we previously showed that myosin II is recruited to the furrow independently of F-actin, and that Rho1 and Rok are essential for this recruitment [1]. Rok phosphorylates several cellular proteins, including the myosin regulatory light chain (RLC).
Methodology/Principal Findings
Here we express phosphorylation state mimic constructs of the RLC in S2 cells to examine the role of RLC phosphorylation involving Rok in the localization of myosin. Phosphorylation of the RLC is required for myosin localization to the equatorial cortex during mitosis, and the essential role of Rok in this localization and for cytokinesis is to maintain phosphorylation of the RLC. The ability to regulate the RLC phosphorylation state spatio-temporally is not essential for the myosin localization. Furthermore, the essential role of Citron in cytokinesis is not phosphorylation of the RLC.
We conclude that the Rho1 pathway leading to myosin localization to the future cytokinetic furrow is relatively straightforward, where only Rok is needed, and it is only needed to maintain phosphorylation of the myosin RLC.
PMCID: PMC1762308  PMID: 17205135
6.  Neighbour-nets portray the Chinese dialect continuum and the linguistic legacy of China's demic history 
As with species studied by evolutionary biologists, languages are evolving entities. They can evolve in tree-like patterns, possibly blurred by borrowing, but they can also develop in non-tree-like schemes. For instance, diglossia, as in the case of Chinese, can counterbalance the hierarchical pattern expected from differentiation by internal change associated with isolation by distance of speech communities. Using two lexical datasets, either the basic lexicon supposedly more immune to borrowing or a representative sample of the whole lexicon, we investigate the development pattern of Chinese dialects using a neighbour-net approach, which is an unprejudiced technique for representing object relationships. The resulting graphs are consistent with a dialect continuum shaped by counterbalanced effects of homogenizing diglossia and borrowing versus differentiating spread of speech communities. Historical events and linguistic claims can be mapped on these graphs.
PMCID: PMC1599877  PMID: 16024359
Chinese; language change; neighbour-net; dialect continuum; diglossia; borrowing
7.  Imaging approach for monitoring cellular metabolites and ions using genetically encoded biosensors 
The spatio-temporal patterns of ion and metabolite levels in living cells are important in understanding signal transduction and metabolite flux. Imaging approaches using genetically encoded sensors are ideal for detecting such molecule dynamics, which are hard to capture otherwise. Recent years have seen iterative improvements and evaluations of sensors, which in turn are starting to make applications in more challenging experimental settings possible. In this review, we will introduce recent progress made in the variety and properties of biosensors, and how biosensors are used for measurement of metabolite and ion in live cells. The emerging field of applications, such as parallel imaging of two separate molecules, high-resolution transport studies and high-throughput screening using biosensors, will be discussed.
PMCID: PMC2843770  PMID: 20167470
8.  Modeling and analysis of mechanisms underlying fMRI-based decoding of information conveyed in cortical columns 
Neuroimage  2010;56(2):627-642.
Multivariate machine learning algorithms applied to human functional MRI (fMRI) data can decode information conveyed by cortical columns, despite the voxel-size being large relative to the width of columns. Several mechanisms have been proposed to underlie decoding of stimulus orientation or the stimulated eye. These include: (I) aliasing of high spatial-frequency components, including the main frequency component of the columnar organization, (II) contributions from local irregularities in the columnar organization, (III) contributions from large-scale non-columnar organizations, (IV) functionally selective veins with biased draining regions, and (V) complex spatio-temporal filtering of neuronal activity by fMRI voxels. Here we sought to assess the plausibility of two of the suggested mechanisms: (I) aliasing and (II) local irregularities, using a naive model of BOLD as blurring and MRI voxel sampling.
To this end, we formulated a mathematical model that encompasses both the processes of imaging ocular dominance (OD) columns and the subsequent linear classification analysis. Through numerical simulations of the model, we evaluated the distribution of functional differential contrasts that can be expected when considering the pattern of cortical columns, the hemodynamic point spread function, the voxel size, and the noise. We found that with data acquisition parameters used at 3 Tesla, sub-voxel supra-Nyquist frequencies, including frequencies near the main frequency of the OD organization (0.5 cycles per mm), cannot contribute to the differential contrast. The differential functional contrast of local origin is dominated by low-amplitude contributions from low frequencies, associated with irregularities of the cortical pattern. Realizations of the model with parameters that reflected best-case scenario and the reported BOLD point-spread at 3 Tesla (3.5 mm) predicted decoding performances lower than those that have been previously obtained at this magnetic field strength. We conclude that low frequency components that underlie local irregularities in the columnar organization are likely to play a role in decoding. We further expect that fMRI-based decoding relies, in part, on signal contributions from large-scale, non-columnar functional organizations, and from complex spatio-temporal filtering of neuronal activity by fMRI voxels, involving biased venous responses. Our model can potentially be used for evaluating and optimizing data-acquisition parameters for decoding information conveyed by cortical columns.
PMCID: PMC3384803  PMID: 20868757
9.  Towards cross-lingual alerting for bursty epidemic events 
Journal of Biomedical Semantics  2011;2(Suppl 5):S10.
Online news reports are increasingly becoming a source for event-based early warning systems that detect natural disasters. Harnessing the massive volume of information available from multilingual newswire presents as many challanges as opportunities due to the patterns of reporting complex spatio-temporal events.
In this article we study the problem of utilising correlated event reports across languages. We track the evolution of 16 disease outbreaks using 5 temporal aberration detection algorithms on text-mined events classified according to disease and outbreak country. Using ProMED reports as a silver standard, comparative analysis of news data for 13 languages over a 129 day trial period showed improved sensitivity, F1 and timeliness across most models using cross-lingual events. We report a detailed case study analysis for Cholera in Angola 2010 which highlights the challenges faced in correlating news events with the silver standard.
The results show that automated health surveillance using multilingual text mining has the potential to turn low value news into high value alerts if informed choices are used to govern the selection of models and data sources. An implementation of the C2 alerting algorithm using multilingual news is available at the BioCaster portal
PMCID: PMC3239300  PMID: 22166371
10.  Phonotactic Diversity Predicts the Time Depth of the World’s Language Families 
PLoS ONE  2013;8(5):e63238.
The ASJP (Automated Similarity Judgment Program) described an automated, lexical similarity-based method for dating the world’s language groups using 52 archaeological, epigraphic and historical calibration date points. The present paper describes a new automated dating method, based on phonotactic diversity. Unlike ASJP, our method does not require any information on the internal classification of a language group. Also, the method can use all the available word lists for a language and its dialects eschewing the debate on ‘language’ vs. ‘dialect’. We further combine these dates and provide a new baseline which, to our knowledge, is the best one. We make a systematic comparison of our method, ASJP’s dating procedure, and combined dates. We predict time depths for world’s language families and sub-families using this new baseline. Finally, we explain our results in the model of language change given by Nettle.
PMCID: PMC3656872  PMID: 23691003
11.  A fluorescent protein biosensor of myosin II regulatory light chain phosphorylation reports a gradient of phosphorylated myosin II in migrating cells. 
Molecular Biology of the Cell  1995;6(12):1755-1768.
Phosphorylation of the regulatory light chain by myosin light chain kinase (MLCK) regulates the motor activity of smooth muscle and nonmuscle myosin II. We have designed reagents to detect this phosphorylation event in living cells. A new fluorescent protein biosensor of myosin II regulatory light chain phosphorylation (FRLC-Rmyosin II) is described here. The biosensor depends upon energy transfer from fluorescein-labeled regulatory light chains to rhodamine-labeled essential and/or heavy chains. The energy transfer ratio increases by up to 26% when the regulatory light chain is phosphorylated by MLCK. The majority of the change in energy transfer is from regulatory light chain phosphorylation by MLCK (versus phosphorylation by protein kinase C). Folding/unfolding, filament assembly, and actin binding do not have a large effect on the energy transfer ratio. FRLC-Rmyosin II has been microinjected into living cells, where it incorporates into stress fibers and transverse fibers. Treatment of fibroblasts containing FRLC-Rmyosin II with the kinase inhibitor staurosporine produced a lower ratio of rhodamine/fluorescein emission, which corresponds to a lower level of myosin II regulatory light chain phosphorylation. Locomoting fibroblasts containing FRLC-Rmyosin II showed a gradient of myosin II phosphorylation that was lowest near the leading edge and highest in the tail region of these cells, which correlates with previously observed gradients of free calcium and calmodulin activation. Maximal myosin II motor force in the tail may contribute to help cells maintain their polarized shape, retract the tail as the cell moves forward, and deliver disassembled subunits to the leading edge for incorporation into new fibers.
PMCID: PMC301330  PMID: 8590803
12.  Using Language Sampling in Clinical Assessments with Bilingual Children: Challenges and Future Directions 
Seminars in Speech and Language  2009;30(4):234-245.
Current language tests designed to assess Spanish-English-speaking children have limited clinical accuracy and do not provide sufficient information to plan language intervention. In contrast, spontaneous language samples obtained in the two languages can help identify language impairment with higher accuracy. In this article, we describe several diagnostic indicators that can be used in language assessments based on spontaneous language samples. First, based on previous research with monolingual and bilingual English speakers, we show that a verb morphology composite measure in combination with a measure of mean length of utterance (MLU) can provide valuable diagnostic information for English development in bilingual children. Dialectal considerations are discussed. Second, we discuss the available research with bilingual Spanish speakers and show a series of procedures to be used for the analysis of Spanish samples: (a) limited MLU and proportional use of ungrammatical utterances; (b) limited grammatical accuracy on articles, verbs, and clitic pronouns; and (c) limited MLU, omission of theme arguments, and limited use of ditransitive verbs. Third, we illustrate the analysis of verb argument structure using a rubric as an assessment tool. Estimated scores on morphological and syntactic measures are expected to increase the sensitivity of clinical assessments with young bilingual children. Further research using other measures of language will be needed for older school-age children.
PMCID: PMC3374634  PMID: 19851951
Bilingual language sample analysis; language disorders; assessment rubic
13.  Bayesian phylogenetic analysis supports an agricultural origin of Japonic languages 
Languages, like genes, evolve by a process of descent with modification. This striking similarity between biological and linguistic evolution allows us to apply phylogenetic methods to explore how languages, as well as the people who speak them, are related to one another through evolutionary history. Language phylogenies constructed with lexical data have so far revealed population expansions of Austronesian, Indo-European and Bantu speakers. However, how robustly a phylogenetic approach can chart the history of language evolution and what language phylogenies reveal about human prehistory must be investigated more thoroughly on a global scale. Here we report a phylogeny of 59 Japonic languages and dialects. We used this phylogeny to estimate time depth of its root and compared it with the time suggested by an agricultural expansion scenario for Japanese origin. In agreement with the scenario, our results indicate that Japonic languages descended from a common ancestor approximately 2182 years ago. Together with archaeological and biological evidence, our results suggest that the first farmers of Japan had a profound impact on the origins of both people and languages. On a broader level, our results are consistent with a theory that agricultural expansion is the principal factor for shaping global linguistic diversity.
PMCID: PMC3203502  PMID: 21543358
linguistic evolution; phylogenetics; Japonic languages
14.  Free classification of American English dialects by native and non-native listeners 
Journal of phonetics  2009;37(4):436-451.
Most second language acquisition research focuses on linguistic structures, and less research has examined the acquisition of sociolinguistic patterns. The current study explored the perceptual classification of regional dialects of American English by native and non-native listeners using a free classification task. Results revealed similar classification strategies for the native and non-native listeners. However, the native listeners were more accurate overall than the non-native listeners. In addition, the non-native listeners were less able to make use of constellations of cues to accurately classify the talkers by dialect. However, the non-native listeners were able to attend to cues that were either phonologically or sociolinguistically relevant in their native language. These results suggest that non-native listeners can use information in the speech signal to classify talkers by regional dialect, but that their lack of signal-independent cultural knowledge about variation in the second language leads to less accurate classification performance.
PMCID: PMC2783622  PMID: 20161400
second language acquisition; dialect classification
15.  Acoustic Processing of Temporally Modulated Sounds in Infants: Evidence from a Combined Near-Infrared Spectroscopy and EEG Study 
Speech perception requires rapid extraction of the linguistic content from the acoustic signal. The ability to efficiently process rapid changes in auditory information is important for decoding speech and thereby crucial during language acquisition. Investigating functional networks of speech perception in infancy might elucidate neuronal ensembles supporting perceptual abilities that gate language acquisition. Interhemispheric specializations for language have been demonstrated in infants. How these asymmetries are shaped by basic temporal acoustic properties is under debate. We recently provided evidence that newborns process non-linguistic sounds sharing temporal features with language in a differential and lateralized fashion. The present study used the same material while measuring brain responses of 6 and 3 month old infants using simultaneous recordings of electroencephalography (EEG) and near-infrared spectroscopy (NIRS). NIRS reveals that the lateralization observed in newborns remains constant over the first months of life. While fast acoustic modulations elicit bilateral neuronal activations, slow modulations lead to right-lateralized responses. Additionally, auditory-evoked potentials and oscillatory EEG responses show differential responses for fast and slow modulations indicating a sensitivity for temporal acoustic variations. Oscillatory responses reveal an effect of development, that is, 6 but not 3 month old infants show stronger theta-band desynchronization for slowly modulated sounds. Whether this developmental effect is due to increasing fine-grained perception for spectrotemporal sounds in general remains speculative. Our findings support the notion that a more general specialization for acoustic properties can be considered the basis for lateralization of speech perception. The results show that concurrent assessment of vascular based imaging and electrophysiological responses have great potential in the research on language acquisition.
PMCID: PMC3110620  PMID: 21716574
infants; speech perception; language acquisition; auditory processing; near-infrared spectroscopy; event related potentials; brain oscillations
16.  Role of Phosphorylation on the Structural Dynamics and Function of Types III and IV Intermediate Filaments 
Experimental cell research  2007;313(10):2098-2109.
Phosphorylation of types III and IV intermediate filaments (IFs) is known to regulate their organization and function. Phosphorylation of the amino-terminal head domain sites on types III and IV IF proteins plays a key role in the assembly/disassembly of IF subunits into 10 nm filaments, and influences the phosphorylation of sites on the carboxyl-terminal tail domain. These phosphorylation events are largely under the control of second messenger-dependent protein kinases and provide the cells a mechanism to reorganize the IFs in response to the changes in second messenger levels. In mitotic cells, Cdk1, Rho kinase, PAK1 and Aurora-B kinase are believed to regulate vimentin and glial fibrillary acidic protein phosphorylation in a spatio-temporal manner. In neurons, the carboxyl-terminal tail domains of the NF-M and NF-H subunits of heteropolymeric neurofilaments (NFs) are highly phosphorylated by proline-directed protein kinases. The phosphorylation of carboxyl-terminal tail domains of NFs has been suspected to play roles in forming cross-bridges between NFs and microtubules, slowing axonal transport and promoting their integration into cytoskeleton lattice and, in doing so, to control axonal caliber and stabilize the axon. The role of IF phosphorylation in disease pathobiology is discussed.
PMCID: PMC2570114  PMID: 17498690
Intermediate filaments; neurofilaments; vimentin; phosphorylation; protein kinases and protein assembly
17.  Animacy or Case Marker Order?: Priority Information for Online Sentence Comprehension in a Head-Final Language 
PLoS ONE  2014;9(3):e93109.
It is well known that case marker information and animacy information are incrementally used to comprehend sentences in head-final languages. However, it is still unclear how these two kinds of information are processed when they are in competition in a sentence's surface expression. The current study used sentences conveying the potentiality of some event (henceforth, potential sentences) in the Japanese language with theoretically canonical word order (dative–nominative/animate–inanimate order) and with scrambled word order (nominative–dative/inanimate–animate order). In Japanese, nominative–first case order and animate–inanimate animacy order are preferred to their reversed patterns in simplex sentences. Hence, in these potential sentences, case information and animacy information are in competition. The experiment consisted of a self-paced reading task testing two conditions (that is, canonical and scrambled potential sentences). Forty-five native speakers of Japanese participated. In our results, the canonical potential sentences showed a scrambling cost at the second argument position (the nominative argument). This result indicates that the theoretically scrambled case marker order (nominative–dative) is processed as a mentally canonical case marker order, suggesting that case information is used preferentially over animacy information when the two are in competition. The implications of our findings are discussed with regard to incremental simplex sentence comprehension models for head-final languages.
PMCID: PMC3963992  PMID: 24664132
18.  Does language guide event perception? Evidence from eye movements 
Cognition  2008;108(1):155-184.
Languages differ in how they encode motion. When describing bounded motion, English speakers typically use verbs that convey information about manner (e.g., slide, skip, walk) rather than path (e.g., approach, ascend), whereas Greek speakers do the opposite. We investigated whether this strong cross-language difference influences how people allocate attention during motion perception. We compared eye movements from Greek and English speakers as they viewed motion events while (a) preparing verbal descriptions, or (b) memorizing the events. During the verbal description task, speakers’ eyes rapidly focused on the event components typically encoded in their native language, generating significant cross-language differences even during the first second of motion onset. However, when freely inspecting ongoing events, as in the memorization task, people allocated attention similarly regardless of the language they speak. Differences between language groups arose only after the motion stopped, such that participants spontaneously studied those aspects of the scene that their language does not routinely encode in verbs. These findings offer a novel perspective on the relation between language and perceptual/cognitive processes. They indicate that attention allocation during event perception is not affected by the perceiver’s native language; effects of language arise only when linguistic forms are recruited to achieve the task, such as when committing facts to memory.
PMCID: PMC2810627  PMID: 18395705
spatial language; language and thought; visual world; eyetracking; motion; Greek; cross-linguistic differences
19.  Fluorescent Biosensors for Real-time Tracking of Post-translational Modification Dynamics 
Dynamic post-translational modifications (PTMs) regulate and diversify protein properties and cellular behaviors. Real-time monitoring of these modifications has been made possible with biosensors based on fluorescent proteins (FPs) and fluorescence resonance energy transfer (FRET), which can provide spatiotemporal information of PTMs with little perturbation to the cellular environment. In this review, we highlight available fluorescent biosensors applicable to detect PTMs in living cells and how they have shed light on biological questions that have been difficult to address otherwise. In addition, we also provide discussions about various engineering strategies for overcoming potential challenges associated with the development and application of such biosensors.
PMCID: PMC2757776  PMID: 19682946
20.  Protein Kinase C: The “Masters” of Calcium and Lipid 
The coordinated and physiological behavior of living cells in an organism critically depends on their ability to interact with surrounding cells and with the extracellular space. For this, cells have to interpret incoming stimuli, correctly process the signals, and produce meaningful responses. A major part of such signaling mechanisms is the translation of incoming stimuli into intracellularly understandable signals, usually represented by second messengers or second-messenger systems. Two key second messengers, namely the calcium ion and signaling lipids, albeit extremely different in nature, play an important and often synergistic role in such signaling cascades. In this report, we will shed some light on an entire family of protein kinases, the protein kinases C, that are perfectly designed to exactly decode these two second messengers in all of their properties and convey the signaling content to downstream processes within the cell.
The kinase domains of protein kinase C lack substrate specificity. Instead, specificity is partly determined by the binding of various lipids and calcium ions to the regulatory domains.
PMCID: PMC3119906  PMID: 21628429
21.  Language in schizophrenia Part 1: an Introduction 
Language and linguistics compass  2010;4(8):576-589.
This is the first of two articles that discuss higher-order language and semantic processing in schizophrenia. This article reviews clinical characterizations of language output and the phenomenon of positive thought disorder, as well as more principled characterizations of language output in schizophrenia. It also gives an overview of evidence for the predominant theory of language dysfunction in schizophrenia: that it arises from abnormalities in (a) semantic memory and/or (b) working memory and executive function. The companion article (Part 2) focuses on the study of language in schizophrenia using online psycholinguistic methods and considers how the study of schizophrenia may inform our understanding of normal language processing.
PMCID: PMC2950318  PMID: 20936080
cognition; context; DLPFC; discourse; disorganization; ERP; fMRI; language; N400; prefrontal cortex; semantic; sentence; schizophrenia; syntax; temporal cortex; thought disorder
22.  Language, gesture, skill: the co-evolutionary foundations of language 
This paper defends a gestural origins hypothesis about the evolution of enhanced communication and language in the hominin lineage. The paper shows that we can develop an incremental model of language evolution on that hypothesis, but not if we suppose that language originated in an expansion of great ape vocalization. On the basis of the gestural origins hypothesis, the paper then advances solutions to four classic problems about the evolution of language: (i) why did language evolve only in the hominin lineage? (ii) why is language use an evolutionarily stable form of informational cooperation, despite the fact that hominins have diverging evolutionary interests? (iii) how did stimulus independent symbols emerge? (iv) what were the origins of complex, syntactically organized symbols? The paper concludes by confronting two challenges: those of testability and of explaining the gesture-to-speech transition; crucial issues for any gestural origins hypothesis
PMCID: PMC3385681  PMID: 22734057
evolution of language; gesture; gestural origins; hominin cooperation; cooperation and communication
23.  A Fluorescent Reporter of AMPK activity and Cellular Energy Stress 
Cell metabolism  2011;13(4):476-486.
AMP-activated protein kinase (AMPK) is activated when the AMP/ATP ratio in cells is elevated due to energy stress. Here we describe a biosensor, AMPKAR, which exhibits enhanced fluorescence resonance energy transfer (FRET) in response to phosphorylation by AMPK, allowing spatio-temporal monitoring of AMPK activity in single cells. We show that this reporter responds to a variety of stimuli that are known to induce energy stress and that the response is dependent on AMPK α1 & α2 and on the upstream kinase, LKB1. Interestingly we found that AMPK activation is confined to the cytosol in response to energy stress but can be observed in both the cytosol and nucleus in response to calcium elevation. Finally, using this probe with U2OS cells in a microfluidics device, we observed a very high cell-to-cell variability in the amplitude and time course of AMPK activation and recovery in response to pulses of glucose deprivation.
PMCID: PMC3070961  PMID: 21459332
24.  Quantifying myosin light chain phosphorylation in single adherent cells with automated fluorescence microscopy 
BMC Cell Biology  2007;8:43.
In anchorage dependent cells, myosin generated contractile forces affect events closely associated with adhesion such as the formation of stress fibers and focal adhesions, and temporally distal events such as entry of the cell into S-phase. As occurs in many signaling pathways, a phosphorylation reaction (in this case, phosphorylation of myosin light chain) is directly responsible for cell response. Western blotting has been useful in measuring intracellular phosphorylation events, but cells are lysed in the process of sample preparation for western blotting, and spatial information such as morphology, localization of the phosphorylated species, and the distribution of individual cell responses across the population is lost. We report here a reliable automated microscopy method for quantitative measurement of myosin light chain phosphorylation in adherent cells. This method allows us to concurrently examine cell morphology, cell-cell contact, and myosin light chain diphosphorylation in vascular smooth muscle cells.
Paraformaldehyde fixation and Triton X-100 permeabilization preserved cell morphology and myosin light chain phosphorylation better than the alternative fixation/permeabilization methods tested. We utilized automated microscopy methods to acquire three color images, determine cell spread area, and quantify the intensity of staining within each cell with anti-phospho-MLC antibody. Our results indicate that A10 rat aortic smooth muscle cells exhibit a re producible non-Gaussian distribution of MLC phosphorylation across a population of unsynchronized genetically identical cells. Adding an inhibitor of Rho kinase, Y27632, or plating cells on a low density of fibronectin, reduced phospho-myosin light chain signal as expected. On the other hand, adding calyculin A, an activator of contractility, increased myosin light chain phosphorylation. The IC50 for myosin light chain phosphorylation using Y27632 was determined to be 2.1 ± 0.6 micrometers. We observed a positive linear relationship between cell area and myosin light chain diphosphorylation, which is consistent with what has been reported in the literature using other methods.
Our results show that using proper specimen fixation techniques and background subtraction methods, imaging cytometry can be used to reliably measure relative myosin light chain phosphorylation in individual adherent cells. Importantly, the ability to make this measurement in adherent cells allows for simultaneous measurement of and correlation with other parameters of cellular topography such as morphology and cell-cell proximity. This assay has potential application in screening for drug development.
PMCID: PMC2213650  PMID: 17941977
25.  Frontal and temporal contributions to understanding the iconic co-speech gestures that accompany speech 
Human brain mapping  2012;35(3):900-917.
In everyday conversation, listeners often rely on a speaker’s gestures to clarify any ambiguities in the verbal message. Using fMRI during naturalistic story comprehension, we examined which brain regions in the listener are sensitive to speakers’ iconic gestures. We focused on iconic gestures that contribute information not found in the speaker’s talk, compared to those that convey information redundant with the speaker’s talk. We found that three regions—left inferior frontal gyrus triangular (IFGTr) and opercular (IFGOp) portions, and left posterior middle temporal gyrus (MTGp)—responded more strongly when gestures added information to non-specific language, compared to when they conveyed the same information in more specific language; in other words, when gesture disambiguated speech as opposed to reinforced it. An increased BOLD response was not found in these regions when the non-specific language was produced without gesture, suggesting that IFGTr, IFGOp, and MTGp are involved in integrating semantic information across gesture and speech. In addition, we found that activity in the posterior superior temporal sulcus (STSp), previously thought to be involved in gesture-speech integration, was not sensitive to the gesture-speech relation. Together, these findings clarify the neurobiology of gesture-speech integration and contribute to an emerging picture of how listeners glean meaning from gestures that accompany speech.
PMCID: PMC3797208  PMID: 23238964
gestures; semantic; language; inferior frontal gyrus; posterior superior temporal sulcus; posterior middle temporal gyrus

Results 1-25 (768367)