Expression and functional significance of phosphoenolpyruvate carboxykinase 1 in uveal melanoma

Abstract Uveal melanoma (UVM), an uncommon yet potentially life-threatening ocular cancer, arises from melanocytes in the uveal tract of the eye. The exploration of novel oncotargets for UVM is of paramount importance. In this study, we show that PCK1 (phosphoenolpyruvate carboxykinase 1) expression is upregulated in various UVM tissues as well as in primary UVM cells and immortalized lines. Furthermore, bioinformatics studies reveal that PCK1 overexpression in UVM correlates with advanced disease stages and poor patient survival. Genetic silencing (utilizing viral shRNA) or knockout (via CRISPR/Cas9) of PCK1 significantly curtailed cell viability, proliferation, cell cycle progression, and motility, while provoking apoptosis in primary and immortalized UVM cells. Conversely, ectopic overexpression of PCK1, achieved through a viral construct, bolstered UVM cell proliferation and migration. Gαi3 expression and Akt phosphorylation were reduced following PCK1 silencing or knockout, but increased after PCK1 overexpression in UVM cells. Restoring Akt phosphorylation through a constitutively active mutant Akt1 (S473D) ameliorated the growth inhibition, migration suppression, and apoptosis induced by PCK1 silencing in UVM cells. Additionally, ectopic expression of Gαi3 restored Akt activation and counteracted the anti-UVM cell effects by PCK1 silencing. In vivo, the growth of subcutaneous xenografts of primary human UVM cells was significantly inhibited following intratumoral injection of adeno-associated virus (aav) expressing PCK1 shRNA. PCK1 depletion, Gαi3 downregulation, Akt inhibition, proliferation arrest, and apoptosis were detected in PCK1-silenced UVM xenografts. Collectively, our findings demonstrate that PCK1 promotes UVM cell growth possibly by modulating the Gαi3-Akt signaling pathway

Long-Term Experience of Chinese Calligraphic Handwriting Is Associated with Better Executive Functions and Stronger Resting-State Functional Connectivity in Related Brain Regions

Chinese calligraphic handwriting (CCH) is a traditional art form that requires high levels of concentration and motor control. Previous research has linked short-term training in CCH to improvements in attention and memory. Little is known about the potential impacts of long-term CCH practice on a broader array of executive functions and their potential neural substrates. In this cross-sectional study, we recruited 36 practitioners with at least 5 years of CCH experience and 50 control subjects with no more than one month of CCH practice and investigated their differences in the three components of executive functions (i.e., shifting, updating, and inhibition). Valid resting-state fMRI data were collected from 31 CCH and 40 control participants. Compared with the controls, CCH individuals showed better updating (as measured by the Corsi Block Test) and inhibition (as measured by the Stroop Word-Color Test), but the two groups did not differ in shifting (as measured by a cue-target task). The CCH group showed stronger resting-state functional connectivity (RSFC) than the control group in brain areas involved in updating and inhibition. These results suggested that long-term CCH training may be associated with improvements in specific aspects of executive functions and strengthened neural networks in related brain regions

Long-term Chinese calligraphic handwriting training has a positive effect on brain network efficiency.

As a visual art form, Chinese calligraphic handwriting (CCH) has been found to correlate with certain brain activity and to induce functional connectivity reorganization of the brain. This study investigated the effect of long-term CCH training on brain functional plasticity as assessed with network measures. With the resting-state fMRI data from 31 participants with at least five years of CCH training and 40 controls, we constructed brain functional networks, examined group differences at both the whole brain and modular levels, and correlated the topological characteristics with calligraphy skills. We found that, compared to the control group, the CCH group showed shorter characteristic path lengths and higher local efficiency in certain brain areas in the frontal and parietal cortices, limbic system, basal ganglia, and thalamus. Moreover, these network measures in the cingulate cortex, caudate nucleus, and thalamus were associated with CCH performance (i.e., copying and creating skills). These results suggest that long-term CCH training has a positive effect on the topological characteristics of brain networks

Long-term Chinese calligraphic handwriting training has a positive effect on brain network efficiency.

As a visual art form, Chinese calligraphic handwriting (CCH) has been found to correlate with certain brain activity and to induce functional connectivity reorganization of the brain. This study investigated the effect of long-term CCH training on brain functional plasticity as assessed with network measures. With the resting-state fMRI data from 31 participants with at least five years of CCH training and 40 controls, we constructed brain functional networks, examined group differences at both the whole brain and modular levels, and correlated the topological characteristics with calligraphy skills. We found that, compared to the control group, the CCH group showed shorter characteristic path lengths and higher local efficiency in certain brain areas in the frontal and parietal cortices, limbic system, basal ganglia, and thalamus. Moreover, these network measures in the cingulate cortex, caudate nucleus, and thalamus were associated with CCH performance (i.e., copying and creating skills). These results suggest that long-term CCH training has a positive effect on the topological characteristics of brain networks

Long-term Chinese calligraphic handwriting training has a positive effect on brain network efficiency.

As a visual art form, Chinese calligraphic handwriting (CCH) has been found to correlate with certain brain activity and to induce functional connectivity reorganization of the brain. This study investigated the effect of long-term CCH training on brain functional plasticity as assessed with network measures. With the resting-state fMRI data from 31 participants with at least five years of CCH training and 40 controls, we constructed brain functional networks, examined group differences at both the whole brain and modular levels, and correlated the topological characteristics with calligraphy skills. We found that, compared to the control group, the CCH group showed shorter characteristic path lengths and higher local efficiency in certain brain areas in the frontal and parietal cortices, limbic system, basal ganglia, and thalamus. Moreover, these network measures in the cingulate cortex, caudate nucleus, and thalamus were associated with CCH performance (i.e., copying and creating skills). These results suggest that long-term CCH training has a positive effect on the topological characteristics of brain networks

Semantic representation in the white matter pathway

<div><p>Object conceptual processing has been localized to distributed cortical regions that represent specific attributes. A challenging question is how object semantic space is formed. We tested a novel framework of representing semantic space in the pattern of white matter (WM) connections by extending the representational similarity analysis (RSA) to structural lesion pattern and behavioral data in 80 brain-damaged patients. For each WM connection, a neural representational dissimilarity matrix (RDM) was computed by first building machine-learning models with the voxel-wise WM lesion patterns as features to predict naming performance of a particular item and then computing the correlation between the predicted naming score and the actual naming score of another item in the testing patients. This correlation was used to build the neural RDM based on the assumption that if the connection pattern contains certain aspects of information shared by the naming processes of these two items, models trained with one item should also predict naming accuracy of the other. Correlating the neural RDM with various cognitive RDMs revealed that neural patterns in several WM connections that connect left occipital/middle temporal regions and anterior temporal regions associated with the object semantic space. Such associations were not attributable to modality-specific attributes (shape, manipulation, color, and motion), to peripheral picture-naming processes (picture visual similarity, phonological similarity), to broad semantic categories, or to the properties of the cortical regions that they connected, which tended to represent multiple modality-specific attributes. That is, the semantic space could be represented through WM connection patterns across cortical regions representing modality-specific attributes.</p></div

WM connections representing higher-order semantic space.

<p>(A) Eight WM connections representing higher-order semantic space, with 11 GM regions being connected. The regions that fail to show successful within-item prediction or in right hemisphere are rendered gray. The four colored regions represent raw semantic effects or modality-specific attributes (red for manipulation, shape, and semantic; orange for manipulation and shape; and purple for shape and color). (B) The WM connections reconstructed using the HCP database. The blue streamlines are the WM connections between two GM regions (rendered in red and green). The masks of WM connections reconstructed with current data are shown in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003993#pbio.2003993.s002" target="_blank">S2 Fig</a>. The RSA results of the eight WM connections, with bars showing the correlation strength (<i>r</i> value) between neural and semantic RDMs and error bars indicating ±1 standard error based on 1,000 times bootstrap resampling (see [<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003993#pbio.2003993.ref023" target="_blank">23</a>] for details) of the neural and behavioral RDM sets. The three WM connections did not survive all validation tests were shown in the dashed box. (C) The GM nodes representing semantic and modality-specific knowledge. The bar figure shows the RSA correlation strength (<i>r</i> value) of the semantic and modality-specific attributes in the colored GM regions in (A); the error bars indicate ±1 standard error; only positive values are shown. Note that for the superior ATL, in which RSA with semantic space was significant, its semantic effects diminished when controlling for modality-specific attribute RDMs. Asterisks indicate FDR <i>q</i> < 0.05. The object line drawings were done by first author Y.F.; the brain illustrations were generated using BrainNet Viewer [<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2003993#pbio.2003993.ref028" target="_blank">28</a>] and DSI Studio (<a href="http://dsi-studio.labsolver.org/" target="_blank">http://dsi-studio.labsolver.org/</a>). The underlying data can be found at <a href="https://osf.io/h7upk/?view_only=52b8f86cffa14ed4844e4a1b9cd429cb" target="_blank">https://osf.io/h7upk/?view_only=52b8f86cffa14ed4844e4a1b9cd429cb</a>. ATL, anterior temporal lobe; CAL, calcarine sulcus; FDR, false discovery rate; GM, gray matter; HCP, Human Connectome Project; LING, lingual gyrus; midATL, middle anterior temporal lobe; MOG, middle occipital gyrus; MTG, middle temporal gyrus; PHG, parahippocampal gyrus; PoCG, postcentral gyrus; RDM, representational dissimilarity matrix; RSA, representational similarity analysis; STG, superior temporal gyrus; supATL, superior anterior temporal lobe; WM, white matter.</p

The RSA results of the WM connections showing significant effects of higher-order semantic space.

<p>For each connection, results (r(<i>p</i>)) are shown for the higher-order semantic space, raw semantic space, and broad object category, before or after controlling for various types of stimulus properties and in various subsets of patients. R values are the Spearman <i>r</i> between the neural RDM in the corresponding connection and the semantic RDM with various other properties controlled for.</p

The construction and result of behavioral RDMs.

<p>(A) The multi-arrangement method. Twenty college students were asked to arrange object pictures according to their semantic (or modality-specific attribute) relatedness by dragging the items on a screen with a mouse. The distances between items on the screen would transform into an RDM. If two items, e.g., scissors and axe, showed a close distance, then they were assigned a low value in the RDM. (B) The results of the behavioral RDMs. Three broad types of distances were measured: semantic similarity, modality-specific attributes (shape, manipulation, color, and motion), and control models that are also potentially relevant to object naming (early visual, phonological, and object category matrix). The values of dissimilarity were transformed to percentile for display. Red indicates low dissimilarity (high similarity) and blue high dissimilarity (low similarity). (C) Visualization of the semantic RDM using multidimensional scaling. (D) The correlations among various behavioral RDMs. The object line drawings were done by the first author Y.F. The underlying data for this figure can be found at <a href="https://osf.io/h7upk/?view_only=52b8f86cffa14ed4844e4a1b9cd429cb" target="_blank">https://osf.io/h7upk/?view_only=52b8f86cffa14ed4844e4a1b9cd429cb</a>. F&V, fruit and vegetable; MDS, multidimensional scaling; RDM, representational dissimilarity matrix.</p