Strain Analysis as a Tool for Functional Morphology

Strain gauge analysis can provide tremendous benefits to functionalmorphologists by accurately assessing the performance of a particularhard tissue structure in the mechanicalloading environmentsin which it functions. This method replaces estimated calculationsof skeletal stresses or strains with empirical measurements,and thus allows explicit testing of predictions derived fromtheoretical mechanical analyses or models. This direct measurementis particularly useful and important when the assumptions ofmechanical models are violated in the biological world, suchas when the geometry of the element of interest is complex and/orwhen the loads applied to a structure are either poorly knownof dynamically changing. In many, perhaps most, systems of interestto functional morphologists, the assumptions of simple shapeand well-defined loading necessary for most theoretical analysesare untenable; in these situations, strain gauge analysis canprovide considerable crucial information not obtainable anyother way. Although there are important limitations to the useof this approach, it can be applied to a variety of kinds offunctional analyses and may supply information about skeletalperformance that can contribute to meaningful functional interpretationsof morphological design.     

Shared Brain Lateralization Patterns in Language and Acheulean Stone Tool Production: A Functional Transcranial Doppler Ultrasound Study

Background

The popular theory that complex tool-making and language co-evolved in the human lineage rests on the hypothesis that both skills share underlying brain processes and systems. However, language and stone tool-making have so far only been studied separately using a range of neuroimaging techniques and diverse paradigms.

Methodology/Principal Findings

We present the first-ever study of brain activation that directly compares active Acheulean tool-making and language. Using functional transcranial Doppler ultrasonography (fTCD), we measured brain blood flow lateralization patterns (hemodynamics) in subjects who performed two tasks designed to isolate the planning component of Acheulean stone tool-making and cued word generation as a language task. We show highly correlated hemodynamics in the initial 10 seconds of task execution.

Conclusions/Significance

Stone tool-making and cued word generation cause common cerebral blood flow lateralization signatures in our participants. This is consistent with a shared neural substrate for prehistoric stone tool-making and language, and is compatible with language evolution theories that posit a co-evolution of language and manual praxis. In turn, our results support the hypothesis that aspects of language might have emerged as early as 1.75 million years ago, with the start of Acheulean technology.     

Functional Morphology as a Tool in General Education

SYNOPSIS. This paper explores the proposition that in morphology,or any other scientific subject, every statement is based onmultiple assumptions already in our minds before we begin toteach or do research in the field. Progress in morphology isintimately related to its philosophical and historical foundations.Personalities, social pressures and cultural traditions alsoplay major roles in the often complex and circuitous routesof scientific discovery. General education provides a foundationfor understanding these many interactions. Morphology, itself,is a metaphor for inter-relatedness, and this metaphor can beused to explore general education in the broadest sense. Morphologyis not only a body of knowledge and processes for producingthat knowledge but also groups of people deciding on directions,collaborating on methods, and consulting on concepts. A morphologistis a member of a community of understanding in a culture composedof diverse and often conflicting influences. I suggest thatexploring these issues is an important pedagogic tool for discoveringand understanding the foundations of morphology as a scientificendeavor. This paper is appropriately dedicated to ProfessorMilton Hildebrand who understands well the interdisciplinarynature of scientific discovery.     

Functional Brain Connectivity as a New Feature for P300 Speller

The brain is a large-scale complex network often referred to as the “connectome”. Cognitive functions and information processing are mainly based on the interactions between distant brain regions. However, most of the ‘feature extraction’ methods used in the context of Brain Computer Interface (BCI) ignored the possible functional relationships between different signals recorded from distinct brain areas. In this paper, the functional connectivity quantified by the phase locking value (PLV) was introduced to characterize the evoked responses (ERPs) obtained in the case of target and non-targets visual stimuli. We also tested the possibility of using the functional connectivity in the context of ‘P300 speller’. The proposed approach was compared to the well-known methods proposed in the state of the art of “P300 Speller”, mainly the peak picking, the area, time/frequency based features, the xDAWN spatial filtering and the stepwise linear discriminant analysis (SWLDA). The electroencephalographic (EEG) signals recorded from ten subjects were analyzed offline. The results indicated that phase synchrony offers relevant information for the classification in a P300 speller. High synchronization between the brain regions was clearly observed during target trials, although no significant synchronization was detected for a non-target trial. The results showed also that phase synchrony provides higher performance than some existing methods for letter classification in a P300 speller principally when large number of trials is available. Finally, we tested the possible combination of both approaches (classical features and phase synchrony). Our findings showed an overall improvement of the performance of the P300-speller when using Peak picking, the area and frequency based features. Similar performances were obtained compared to xDAWN and SWLDA when using large number of trials.     

Modularity of Protein Folds as a Tool for Template-Free Modeling of Structures

Predicting the three-dimensional structure of proteins from their amino acid sequences remains a challenging problem in molecular biology. While the current structural coverage of proteins is almost exclusively provided by template-based techniques, the modeling of the rest of the protein sequences increasingly require template-free methods. However, template-free modeling methods are much less reliable and are usually applicable for smaller proteins, leaving much space for improvement. We present here a novel computational method that uses a library of supersecondary structure fragments, known as Smotifs, to model protein structures. The library of Smotifs has saturated over time, providing a theoretical foundation for efficient modeling. The method relies on weak sequence signals from remotely related protein structures to create a library of Smotif fragments specific to the target protein sequence. This Smotif library is exploited in a fragment assembly protocol to sample decoys, which are assessed by a composite scoring function. Since the Smotif fragments are larger in size compared to the ones used in other fragment-based methods, the proposed modeling algorithm, SmotifTF, can employ an exhaustive sampling during decoy assembly. SmotifTF successfully predicts the overall fold of the target proteins in about 50% of the test cases and performs competitively when compared to other state of the art prediction methods, especially when sequence signal to remote homologs is diminishing. Smotif-based modeling is complementary to current prediction methods and provides a promising direction in addressing the structure prediction problem, especially when targeting larger proteins for modeling.     

A Study of Fatty Acids as a Taxonomic Tool for Dermatophyte Fungi

The fatty acid composition of 64 strains from 15 species of dermatophyte fungi was studied by gas-liquid chromatography. The percentage amounts of palmitic, palmitoleic, stearic, oleic and linoleic acids were measured. These proved to be very similar for all species, with considerable intraspecific variation. As a group, Epidermophyton floccosum strains could be distinguished but individual strains could not be assigned reliably to any species or genus. No significant changes due to age or formaldehyde treatment were noted.     

BrainNet Viewer: A Network Visualization Tool for Human Brain Connectomics

The human brain is a complex system whose topological organization can be represented using connectomics. Recent studies have shown that human connectomes can be constructed using various neuroimaging technologies and further characterized using sophisticated analytic strategies, such as graph theory. These methods reveal the intriguing topological architectures of human brain networks in healthy populations and explore the changes throughout normal development and aging and under various pathological conditions. However, given the huge complexity of this methodology, toolboxes for graph-based network visualization are still lacking. Here, using MATLAB with a graphical user interface (GUI), we developed a graph-theoretical network visualization toolbox, called BrainNet Viewer, to illustrate human connectomes as ball-and-stick models. Within this toolbox, several combinations of defined files with connectome information can be loaded to display different combinations of brain surface, nodes and edges. In addition, display properties, such as the color and size of network elements or the layout of the figure, can be adjusted within a comprehensive but easy-to-use settings panel. Moreover, BrainNet Viewer draws the brain surface, nodes and edges in sequence and displays brain networks in multiple views, as required by the user. The figure can be manipulated with certain interaction functions to display more detailed information. Furthermore, the figures can be exported as commonly used image file formats or demonstration video for further use. BrainNet Viewer helps researchers to visualize brain networks in an easy, flexible and quick manner, and this software is freely available on the NITRC website (www.nitrc.org/projects/bnv/).     

Functional Characterization of IPSC-Derived Brain Cells as a Model for X-Linked Adrenoleukodystrophy

X-ALD is an inherited neurodegenerative disorder where mutations in the ABCD1 gene result in clinically diverse phenotypes: the fatal disorder of cerebral childhood ALD (cALD) or a milder disorder of adrenomyeloneuropathy (AMN). The various models used to study the pathobiology of X-ALD disease lack the appropriate presentation for different phenotypes of cALD vs AMN. This study demonstrates that induced pluripotent stem cells (IPSC) derived brain cells astrocytes (Ast), neurons and oligodendrocytes (OLs) express morphological and functional activities of the respective brain cell types. The excessive accumulation of saturated VLCFA, a “hallmark” of X-ALD, was observed in both AMN OLs and cALD OLs with higher levels observed in cALD OLs than AMN OLs. The levels of ELOVL1 (ELOVL Fatty Acid Elongase 1) mRNA parallel the VLCFA load in AMN and cALD OLs. Furthermore, cALD Ast expressed higher levels of proinflammatory cytokines than AMN Ast and control Ast with or without stimulation with lipopolysaccharide. These results document that IPSC-derived Ast and OLs from cALD and AMN fibroblasts mimic the respective biochemical disease phenotypes and thus provide an ideal platform to investigate the mechanism of VLCFA load in cALD OLs and VLCFA-induced inflammatory disease mechanisms of cALD Ast and thus for testing of new therapeutics for AMN and cALD disease of X-ALD.     

Drosophila Lethal Giant Larvae Neoplastic Mutant as a Genetic Tool for Cancer Modeling

Drosophila lethal giant larvae (lgl) is a tumour suppressor gene whose function in establishing apical-basal cell polarity as well as in exerting proliferation control in epithelial tissues is conserved between flies and mammals. Individuals bearing lgl null mutations show a gradual loss of tissue architecture and an extended larval life in which cell proliferation never ceases and no differentiation occurs, resulting in prepupal lethality. When tissues from those individuals are transplanted into adult normal recipients, a subset of cells, possibly the cancer stem units, are again able to proliferate and give rise to metastases which migrate to distant sites killing the host. This phenotype closely resembles that of mammalian epithelial cancers, in which loss of cell polarity is one of the hallmarks of a malignant, metastatic behaviour associated with poor prognosis. Lgl protein shares with its human counterpart Human giant larvae-1 (Hugl-1) significant stretches of sequence similarity that we demonstrated to translate into a complete functional conservation, pointing out a role in cell proliferation control and tumorigenesis also for the human homologue. The functional conservation and the power of fly genetics, that allows the researcher to manipulate the fly genome at a level of precision that exceeds that of any other multicellular genetic system, make this Drosophila mutant a very suitable model in which to investigate the mechanisms underlying epithelial tumour formation, progression and metastatisation. In this review, we will summarise the results obtained in these later years using this model for the study of cancer biology. Moreover, we will discuss how recent advances in developmental genetics techniques have succeeded in enhancing the similarities between fly and human tumorigenesis, giving Drosophila a pivotal role in the study of such a complex genetic disease.Key Words: Lethal giant larvae, tumour suppressor, Drosophila, Hugl-1, epithelial cancers, animal model.     

A Model of Functional Brain Connectivity and Background Noise as a Biomarker for Cognitive Phenotypes: Application to Autism

We present an efficient approach to discriminate between typical and atypical brains from macroscopic neural dynamics recorded as magnetoencephalograms (MEG). Our approach is based on the fact that spontaneous brain activity can be accurately described with stochastic dynamics, as a multivariate Ornstein-Uhlenbeck process (mOUP). By fitting the data to a mOUP we obtain: 1) the functional connectivity matrix, corresponding to the drift operator, and 2) the traces of background stochastic activity (noise) driving the brain. We applied this method to investigate functional connectivity and background noise in juvenile patients (n = 9) with Asperger’s syndrome, a form of autism spectrum disorder (ASD), and compared them to age-matched juvenile control subjects (n = 10). Our analysis reveals significant alterations in both functional brain connectivity and background noise in ASD patients. The dominant connectivity change in ASD relative to control shows enhanced functional excitation from occipital to frontal areas along a parasagittal axis. Background noise in ASD patients is spatially correlated over wide areas, as opposed to control, where areas driven by correlated noise form smaller patches. An analysis of the spatial complexity reveals that it is significantly lower in ASD subjects. Although the detailed physiological mechanisms underlying these alterations cannot be determined from macroscopic brain recordings, we speculate that enhanced occipital-frontal excitation may result from changes in white matter density in ASD, as suggested in previous studies. We also venture that long-range spatial correlations in the background noise may result from less specificity (or more promiscuity) of thalamo-cortical projections. All the calculations involved in our analysis are highly efficient and outperform other algorithms to discriminate typical and atypical brains with a comparable level of accuracy. Altogether our results demonstrate a promising potential of our approach as an efficient biomarker for altered brain dynamics associated with a cognitive phenotype.     

Virus-Induced Gene Silencing as a Tool for Comparative Functional Studies in Thalictrum

Perennial woodland herbs in the genus Thalictrum exhibit high diversity of floral morphology, including four breeding and two pollination systems. Their phylogenetic position, in the early-diverging eudicots, makes them especially suitable for exploring the evolution of floral traits and the fate of gene paralogs that may have shaped the radiation of the eudicots. A current limitation in evolution of plant development studies is the lack of genetic tools for conducting functional assays in key taxa spanning the angiosperm phylogeny. We first show that virus-induced gene silencing (VIGS) of a PHYTOENE DESATURASE ortholog (TdPDS) can be achieved in Thalictrum dioicum with an efficiency of 42% and a survival rate of 97%, using tobacco rattle virus (TRV) vectors. The photobleached leaf phenotype of silenced plants significantly correlates with the down-regulation of endogenous TdPDS (P<0.05), as compared to controls. Floral silencing of PDS was achieved in the faster flowering spring ephemeral T. thalictroides. In its close relative, T. clavatum, silencing of the floral MADS box gene AGAMOUS (AG) resulted in strong homeotic conversions of floral organs. In conclusion, we set forth our optimized protocol for VIGS by vacuum-infiltration of Thalictrum seedlings or dormant tubers as a reference for the research community. The three species reported here span the range of floral morphologies and pollination syndromes present in Thalictrum. The evidence presented on floral silencing of orthologs of the marker gene PDS and the floral homeotic gene AG will enable a comparative approach to the study of the evolution of flower development in this group.     

Disrupted Brain Functional Network in Internet Addiction Disorder: A Resting-State Functional Magnetic Resonance Imaging Study

Internet addiction disorder (IAD) is increasingly recognized as a mental health disorder, particularly among adolescents. The pathogenesis associated with IAD, however, remains unclear. In this study, we aim to explore the encephalic functional characteristics of IAD adolescents at rest using functional magnetic resonance imaging data. We adopted a graph-theoretic approach to investigate possible disruptions of functional connectivity in terms of network properties including small-worldness, efficiency, and nodal centrality on 17 adolescents with IAD and 16 socio-demographically matched healthy controls. False discovery rate-corrected parametric tests were performed to evaluate the statistical significance of group-level network topological differences. In addition, a correlation analysis was performed to assess the relationships between functional connectivity and clinical measures in the IAD group. Our results demonstrate that there is significant disruption in the functional connectome of IAD patients, particularly between regions located in the frontal, occipital, and parietal lobes. The affected connections are long-range and inter-hemispheric connections. Although significant alterations are observed for regional nodal metrics, there is no difference in global network topology between IAD and healthy groups. In addition, correlation analysis demonstrates that the observed regional abnormalities are correlated with the IAD severity and behavioral clinical assessments. Our findings, which are relatively consistent between anatomically and functionally defined atlases, suggest that IAD causes disruptions of functional connectivity and, importantly, that such disruptions might link to behavioral impairments.     

CRISPR/Cas9 as Tool for Functional Study of Genes Involved in Preimplantation Embryo Development

The CRISPR/Cas9 system has proven to be an efficient gene-editing tool for genome modification of cells and organisms. However, the applicability and efficiency of this system in pig embryos have not been studied in depth. Here, we aimed to remove porcine OCT4 function as a model case using the CRISPR/Cas9 system. Injection of Cas9 and single-guide RNA (sgRNA) against OCT4 decreased the percentages of OCT4-positive embryos to 37–50% of total embryos, while ~100% of control embryos exhibited clear OCT4 immunostaining. We assessed the mutation status near the guide sequence using polymerase chain reaction (PCR) and DNA sequencing, and a portion of blastocysts (20% in exon 2 and 50% in exon 5) had insertions/deletions near protospacer-adjacent motifs (PAMs). Different target sites had frequent deletions, but different concentrations of sgRNA made no impact. OCT4 mRNA levels dramatically decreased at the 8-cell stage, and they were barely detectable in blastocysts, while mRNA levels of other genes, including NANOG, and CDX2 were not affected. In addition, the combination of two sgRNAs led to large-scale deletion (about 1.8 kb) in the same chromosome. Next, we injected an enhanced green fluorescent protein (eGFP) vector targeting the OCT4 exon with Cas9 and sgRNA to create a knockin. We confirmed eGFP fluorescence in blastocysts in the inner cell mass, and also checked the mutation status using PCR and DNA sequencing. A significant portion of blastocysts had eGFP sequence insertions near PAM sites. The CRISPR/CAS9 system provides a good tool for gene functional studies by deleting target genes in the pig.     

Evaluating Amazon's Mechanical Turk as a Tool for Experimental Behavioral Research

Amazon Mechanical Turk (AMT) is an online crowdsourcing service where anonymous online workers complete web-based tasks for small sums of money. The service has attracted attention from experimental psychologists interested in gathering human subject data more efficiently. However, relative to traditional laboratory studies, many aspects of the testing environment are not under the experimenter''s control. In this paper, we attempt to empirically evaluate the fidelity of the AMT system for use in cognitive behavioral experiments. These types of experiment differ from simple surveys in that they require multiple trials, sustained attention from participants, comprehension of complex instructions, and millisecond accuracy for response recording and stimulus presentation. We replicate a diverse body of tasks from experimental psychology including the Stroop, Switching, Flanker, Simon, Posner Cuing, attentional blink, subliminal priming, and category learning tasks using participants recruited using AMT. While most of replications were qualitatively successful and validated the approach of collecting data anonymously online using a web-browser, others revealed disparity between laboratory results and online results. A number of important lessons were encountered in the process of conducting these replications that should be of value to other researchers.