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The finite penetration depth of light in biological tissues is a practical constraint in light‐induced therapies, such as antimicrobial light therapy, photothermal therapy, and photodynamic cancer therapy. Herein, a biocompatible and implantable device, termed hydrogel planar waveguide with lens‐microneedles, for light delivery in deep tissue is demonstrated. The prototype device, integrated planar waveguide and lens‐microneedles, is fabricated by press‐molding polyethylene glycol diacrylate polymers. The optical beams through the lens‐microneedles are focused at a specific point to realize the optimal intensity profile in the tissue. The adequate treatment depth and region for the hydrogel planar waveguide with five lens‐microneedles are extended to 24 mm and 3.1 cm2. The photoswitchable chemotherapeutic against colorectal cancer cells is switched by using different hydrogel waveguides. The performances of hydrogel‐waveguide‐enabled photoswitching are characterized by the dose responses from the optical microscope, crystal violet staining, and MTT assays. The anticancer drug activated by the hydrogel planar waveguide with five lens‐microneedles is shown to be twice as effective as the other fibers and waveguides in causing cancer cell death. The proposed biodegradable waveguide can be utilized for long‐term light delivery and does not require to be removed as it is gradually resorbed by the tissue. The results point to a new paradigm for widespread use in photomedicine.

Abstract Background Microbiota-gut-brain axis interacts with one another to regulate brain functions. However, whether the impacts of gut dysbiosis on limbic white matter (WM) tracts contribute to the neuropsychiatric symptoms (NPS) in patients with amyloid-positive amnestic mild cognitive impairment (aMCI+), have not been explored yet. This study aimed to investigate the mediation effects of limbic WM integrity on the association between gut microbiota and NPS in patients with aMCI+. Methods Twenty patients with aMCI + and 20 healthy controls (HCs) were enrolled. All subjects underwent neuropsychological assessments and their microbial compositions were characterized using 16S rRNA Miseq sequencing technique. Amyloid deposition inspected by positron emission tomography imaging and limbic WM tracts (i.e., fornix, cingulum, and uncinate fasciculus) detected by diffusion tensor imaging were additionally measured in patients with aMCI+. We employed a regression-based mediation analysis using Hayes’s PROCESS macro in this study. Results The relative abundance of genera Ruminococcus and Lactococcus was significantly decreased in patients with aMCI + versus HCs. The relative abundance of Ruminococcus was negatively correlated with affective symptom cluster in the aMCI + group. Notably, this association was mediated by WM integrity of the left cingulate gyrus. Conclusions Our findings suggest Ruminococcus as a potential target for the management of affective impairments in patients with aMCI+.

Background Neural gating of respiratory sensations (NGRS) characterises the brain's ability to filter out repetitive respiratory sensory stimuli. This mechanism plays a crucial role in the neural processing of respiratory stimuli. However, whether ageing affects NGRS in healthy adults is still unclear. Therefore, we aimed to measure the effect of age on NGRS as well as the corresponding S1 and S2 components of the respiratory-related evoked potentials (RREPs). Methods Three age groups of healthy adults participated in this study: a young group (YG; age 20–39 years), a middle-aged group (MG; age 40–59 years) and an old group (OG; age ≥60 years). NGRS was measured by the RREPs in the electroencephalogram in response to short-paired respiratory occlusion stimuli (S1 and S2). The S2/S1 ratio of the RREP N1 amplitude (the negative deflection of the RREP at ∼85–135 ms) was used to characterise NGRS. Results The results showed a significantly smaller N1 S2/S1 ratio in the YG than in the MG (p=0.01) and OG (p=0.03). Further analysis showed that the S1 N1 amplitude was larger for the YG compared with the MG (p=0.03) and OG (p=0.007). Moreover, age was significantly correlated with the N1 S2/S1 ratio (r=0.43), with higher age relating to higher N1 S2/S1 ratios. Conclusions The greater N1 S2/S1 ratios observed in older adults suggest that ageing has a negative impact on the NGRS. This might contribute to increased experiences of respiratory sensations such as dyspnoea in ageing adults.

Zebrafish can be used as a phenotype-based fast-screening platform for the discovery of skin-whitening products by observing the formation of melanin in vivo through a dissecting microscope. In this study, we first evaluated commercial whitening ingredients to confirm the potency of the zebrafish model. The zebrafish screening system verified that verbascoside and 5 other commonly used commercial skin-whitening compounds– including sodium ascorbate, azelaic acid, transamin, kojic acid, and PTU- significantly reduce melanin content in a dose-dependent manner. We subsequently conducted preliminary screening of 123 natural sources to identify those with whitening property. Through a two-step screening process, anti-melanogenesis effects of 4 and 9 candidates at low and high concentration were verified respectively. The toxicity, mortality, and malformation of zebrafish embryo were also evaluated. We further investigated the components of C. manghas L. and found 6 major compounds, one of which was manghaslin, a compound known for its melanin-suppressing effect on zebrafish without cytotoxicity. With these properties, manghaslin is a promising candidate as a skin lightening agent or a treatment for melanoma.

Background: Macrophages play crucial roles in immune responses during the course of schistosomal infections. Methods: We currently investigated influence of immunocompetent changes in macrophages via microarray-based analysis, mRNA expression analysis, detection of serum cytokines, and subsequent evaluation of the immune phenotypes following the differentiation of infection-induced lymphocytes in a unique T1/T2 double-transgenic mouse model. Results: The gradual upregulation of genes encoding YM1, YM2, and interleukin (IL)-4/IL-13 receptors in infected mice indicated the role of type 2 alternatively activated macrophages (M2, AAMφs) in immune responses after Schistosoma japonicum egg production. FACS analysis showed that surface markers MHC class II (IA/IE) and CD8α+ of the macrophages also exhibited a dramatic change at the various time points before and after egg-production. The transgenic mouse experiments further demonstrated that the shifting of macrophage phenotypes influenced the percentage of helper T (Th)-2 cells, which was observed to be higher than that of Th1 cells, which increased only at 3 and 5 weeks post-infection. The differentiation of effector B cells showed a similar but more significant trend toward type-2 immunity. Conclusion: These results suggest that the infection of mice with S. japonicum resulted in a final Th2- and Be2-skewed immune response. This may be due to phenotypic changes in the macrophages. The influence of alternatively activated macrophages was also activated by S. japonicum egg production. This study elucidated the existence of variations in immune mechanisms at the schistosome infection stages.

Psychological challenges have been found to impact respiratory symptom perception in healthy individuals as well as in patients with various neurological disorders. Human respiratory sensory gating is an objective measure to examine respiratory sensory information processing of repetitive respiratory mechanical stimuli in the central nervous system. With this electrophysiological method, patients with higher anxiety levels showed reduced respiratory sensory gating function in the cortex, and increased symptom perception. In addition, positive emotional contexts were found to increase the respiratory sensory gating function using RREPs. However, neural substrates related to emotional impacts on respiratory sensory gating remain still unclear. In the present study, we examined the emotion processing of respiratory sensory gating using functional magnetic resonance imaging. We hypothesized that positive compared with neutral stimuli would result in reduced brain activations in cortical areas with the paired occlusion paradigm. Thirty-five healthy adults participated in this event-designed fMRI experiment. Paired inspiratory occlusions (two transient occlusions with a 500 ms inter-stimulus-interval are delivered during one inspiration) were provided using an external trigger outside of the scanner. At least 40 paired inspiratory occlusions were collected for each trial. The experiment contained three runs during which participants underwent 12 min for the paired inspiratory occlusion paradigm while watching a fixation cross (the control condition), neutral and positive emotional picture series. The order of emotional picture series was randomized across the participants. Our results revealed an overall trend of reduction of brain activity from the neutral (minus fixation) condition, to the pleasant (minus fixation) condition. For bilateral thalamus and primary visual cortices, there was no significant difference in neural activation between the two contrasts of pleasant (ContrastP–F) and neutral condition (ContrastN–F). The activation of the mid-cingulate and the orbitofrontal cortex was lower in ContrastP–F compared to ContrastN–F. In conclusion, our results suggest that emotional context, especially positive valence, modulates neural correlates in middle cingulate cortex and orbitofrontal cortex in terms of respiratory sensory gating. Future studies are recommended to test emotional impacts on respiratory sensations in patients with neurological disorders.

In this study, a marine brown alga Sargassum cristaefolium-derived fungal strain, Penicillium sumatraense SC29, was isolated and identified. Column chromatography of the extracts from liquid fermented products of the fungal strain was carried out and led to the isolation of six compounds. Their structures were elucidated by spectroscopic analysis and supported by single-crystal X-ray diffraction as four previously undescribed (R)-3-hydroxybutyric acid and glycolic acid derivatives, namely penisterines A (1) and C–E (3–5) and penisterine A methyl ether (2), isolated for the first time from natural resources, along with (R)-3-hydroxybutyric acid (6). Of these compounds identified, penisterine E (5) was a unique 6/6/6-tricyclic ether with an acetal and two hemiketal functionalities. All the isolates were subjected to in vitro anti-angiogenic assays using a human endothelial progenitor cell (EPCs) platform. Among these, penisterine D (4) inhibited EPC growth, migration, and tube formation without any cytotoxic effect. Further, in in vivo bioassays, the percentages of angiogenesis of compound 3 on Tg (fli1:EGFP) transgenic zebrafish were 54% and 37% as the treated concentration increased from 10.2 to 20.4 µg/mL, respectively, and the percentages of angiogenesis of compound 4 were 52% and 41% as the treated concentration increased from 8.6 to 17.2 µg/mL, respectively. The anti-angiogenic activity of penisterine D (4) makes it an attractive candidate for further preclinical investigation.

Subjective cognitive decline (SCD), a self-reported worsening in cognition concurrent with normal performance on standardized neuropsychological tests, has gained much attention due to its high risks in the development of mild cognitive impairments or Alzheimer’s disease. The existing cross-sectional diffusion tensor imaging (DTI) studies in SCD have shown extremely controversial findings. Furthermore, all of these studies investigated diffusion properties within the voxel, such as fractional anisotropy, mean diffusivity, or axial diffusivity (DA). However, it remains unclear whether individuals with SCD demonstrate alterations of diffusion profile between voxels and their neighbors, as indexed by local diffusion homogeneity (LDH). We selected 30 healthy controls (HCs) and 23 SCD subjects to acquire their whole-brain DTI. Diffusion images were compared using the tract-based spatial statistics method. Diffusion indices with significant between-group tract clusters were extracted from each individual for further region-of-interest (ROI)-based comparisons. Our results showed that subjects with SCD demonstrated reduced LDH in the left superior frontal gyrus (SFG) and DA in the right anterior cingulate cortex compared with the HC group. In contrast, the SCD group showed higher LDH values in the left lingual gyrus (LG) compared with the HC group. Notably, LDH in the left SFG was significantly and negatively correlated with LDH in the left LG. In conclusion, white matter (WM) integrity in the left SFG, right ACC, and left LG is altered in SCD, suggesting that individuals with SCD exhibit detectable changes in WM tracts before they demonstrate objective cognitive deficits.

The present study aimed to use event-related potentials with the stop-signal task to investigate the effects of trait anxiety on inhibitory control, error monitoring, and post-error adjustments. The stop-signal reaction time (SSRT) was used to evaluate the behavioral competence of inhibitory control. Electrophysiological signals of error-related negativity (ERN) and error positivity (Pe) were used to study error perception and error awareness, respectively. Post-error slowing (PES) was applied to examine the behavioral adjustments after making errors. The results showed that SSRT and PES did not differ significantly between individuals with high trait anxiety (HTA) and those with low trait anxiety (LTA). However, individuals with HTA demonstrated reduced ERN amplitudes and prolonged Pe latencies than those with LTA. Prolonged Pe latencies were also significantly associated with poorer post-error adjustments. In conclusion, HTA led to reduced cortical responses to error monitoring. Furthermore, inefficient conscious awareness of errors might lead to maladaptive post-error adjustments.

Background: Music-based intervention has been used as first-line non-pharmacological treatment to improve cognitive function for people with mild cognitive impairment (MCI) or dementia in clinical practice. However, evidence regarding the effect of music-based intervention on general cognitive function as well as subdomains of cognitive functions in these individuals is scarce. Objective: To evaluate the efficacy of music-based interventions on a wide range of cognitive functions in people with MCI or dementia. Method: We searched the effect of various music therapies using randomized controlled trials on cognitive function using several databases. Studies based on any type of dementia or MCI were combined. The effects of music-based intervention on each cognitive function were pooled by meta-analysis. Results: A total of 19 studies involving n = 1024 participants (mean age ranged from 60 to 87 years old) were included. We found statistically significant improvements in MMSE (general cognitive function), the Frontal Assessment Battery (executive function), and the Auditory Verbal Learning Test (episodic memory). Conclusions: This study provides positive evidence to support music-based interventions for improving a wide range of cognitive functions in older adults with MCI and dementia. Therefore, we recommend increased use of music in people’s homes, day care centers and nursing homes. This study was registered with PROSPERO, number 250383.

Background: The study aimed to evaluate the effects of transcranial direct current stimulation (tDCS) on cognition, mood disturbance, pain, and fatigue in people with multiple sclerosis (PwMS).Methods: A literature search was performed on articles published between January 1990 and May 2020 in Pubmed, Medline, and Web of Science using the following keywords and their abbreviation in combinations: multiple sclerosis and transcranial direct current stimulation. Mean effect size (ES) and 95% confidence interval were calculated for each domain of interest.Results: Seventeen articles with a total of 383 PwMS were included in this analysis. For cognition, a strong effect size was found for the trial administering the Symbol Digit Modalities Test (ES: 1.15), whereas trials applying the Attention Network Test showed a negative effect size of −0.49. Moderate to strong effect sizes were observed for mood disturbance (mean ES: 0.92), pain (mean ES: 0.59), and fatigue (mean ES: 0.60). Further subgroup analyses for MS-related fatigue showed that both high and low intensities of stimulation lead to nearly the same degree of favorable effects. More pronounced effects were observed in studies administering the Fatigue Severity Scale compared with studies using other fatigue measures such as the Modified Fatigue Impact Scale.Conclusion: These results provide preliminary evidence that tDCS has a favorable effect on cognitive processing speed, mood disturbance, pain, and fatigue in MS. However, the effects on cognition and fatigue vary based on the specific assessment used.

Airway obstruction activates mechanoreceptors that project to the cerebral cortices in humans, as evidenced by scalp encephalography recordings of cortical neuronal activation, i.e., respiratory-related evoked potential (RREP). However, neural evidence of both high spatial and temporal resolution of occlusion-elicited cortical activation in healthy individuals is lacking. In the present study, we tested our hypothesis that inspiratory mechanical stimuli elicit neural activation in cortical structures that can be recorded using magnetoencephalography (MEG). We further examined the relationship between depression and respiratory symptoms and hemispheric dominance in terms of emotional states. A total of 14 healthy nonsmoking participants completed a respiratory symptom questionnaire and a depression symptom questionnaire, followed by MEG and RREP recordings of inspiratory occlusion. Transient inspiratory occlusion of 300 ms was provided randomly every 2 to 4 breaths, and approximately 80 occlusions were collected in every study participant. Participants were required to press a button for detection when they sensed occlusion. Respiratory-related evoked fields (RREFs) and RREP peaks were identified in terms of latencies and amplitudes in the right and left hemispheres. The Wilcoxon signed-rank test was further used to examine differences in peak amplitudes between the right and left hemispheres. Our results showed that inspiratory occlusion elicited RREF M1 peaks between 80 and 100 ms after triggering. Corresponding neuromagnetic responses peaked in the sensorimotor cortex, insular cortex, lateral frontal cortex, and middle frontal cortex. Overall, the RREF M1 peak amplitude in the right insula was significantly higher than that in the left insula (p = 0.038). The RREP data also showed a trend of higher N1 peak amplitudes in the right hemisphere compared to the left (p = 0.064, one-tailed). Subgroup analysis revealed that the laterality index of sensorimotor cortex activation was significantly different between higher- and lower-depressed individuals (−0.33 vs. −0.02, respectively; p = 0.028). For subjective ratings, a significant relationship was found between an individual’s depression level and their respiratory symptoms (Spearman’s rho = 0.54, p = 0.028, one-tailed). In summary, our results demonstrated that the inspiratory occlusion paradigm is feasible to elicit an RREF M1 peak with MEG. Our imaging results showed that cortical neurons were activated in the sensorimotor, frontal, middle temporal, and insular cortices for the M1 peak. Respiratory occlusion elicited higher cortical neuronal activation in the right insula compared to the left, with a higher tendency for right laterality in the sensorimotor cortex for higher-depressed rather than lower-depressed individuals. Higher levels of depression were associated with higher levels of respiratory symptoms. Future research with a larger sample size is recommended to investigate the role of emotion and laterality in cerebral neural processing of respiratory sensation.

Patients with amnestic mild cognitive impairment (aMCI) demonstrate significant cognitive deficits, especially in the memory aspect. The memory deficiency might be attributed to the difficulties in the inhibitory function to suppress redundant stimuli. Sensory gating (SG) refers to the attenuation of neural responses to the second identical stimulus in a paired-click paradigm, in which auditory stimuli are delivered in pairs with inter-stimulus intervals (ISI) of 500 ms and inter-pair intervals of 6–8 s. It is considered as an electrophysiological signal to reflect the brain’s automatic response to gate out repetitive sensory inputs. However, there has been no study systematically investigating SG function in aMCI patients. Thus, the present study used magnetoencephalography (MEG) to record neuromagnetic responses to a paired-click paradigm in 23 healthy controls (HC) and 26 aMCI patients. The Stimulus 2/Stimulus 1 (S2/S1) amplitude ratio was used to represent the SG function. Compared to HC, aMCI patients showed M50 SG deficits in the left inferior frontal gyrus (IFG) and right inferior parietal lobule (IPL). M100 SG defects were also observed in the right IPL. Based on the ROIs showing significant between-group SG differences, we found that a more deficient M50 SG function in the right IPL was associated with poorer performance in the immediate recall of Logic Memory (LM), Chinese Version Verbal Learning Test (CVVLT) and Digit Span Backward (DSB) Test. Furthermore, the M50 SG ratios of the right IPL together with the neuropsychological performance of LM and CVVLT demonstrated very good accuracy in the discrimination of aMCI from HC. In conclusion, compared to HC, aMCI patients showed a significant SG deficit in the right IPL, which was correlated with the auditory short-term memory function. We suggest the combination of SG in the right IPL, LM and CVVLT to be sensitive indicators to differentiate aMCI patients from HC.

We differentiated the influence of mirror-induced visual conflicts on the perceptual–attention–motor control process by examining the variation of primary motor cortex (M1) activities and the functional connectivity among five brain regions associated with perceptual, motor, and attentional processes. Magnetoencephalography (MEG) was recorded under three conditions: both hands kept stationary with the forearms supinated (resting condition), in-phase bimanual movements with congruent visual feedback [symmetry (Sym) condition], and out-of-phase bimanual movements with incongruent visual feedback [asymmetry (Asy) condition]. We found that compared with the resting state, the decrease in beta oscillation was greater in the Sym than in the Asy condition, suggesting a greater activation of M1 when implementing hand movement without visual conflict. The results of functional connectivity patterns showed that the alpha band functional connectivity between V1 and superior temporal gyrus (STG) and the gamma band functional connectivity between the precuneus and posterior cingulate cortex (PCC) triggered greater or slightly greater coherence strength in the Asy condition than in the Sym condition. However, the beta band functional connectivity showed no difference between the two conditions in all pairs of the brain regions. These findings confirm and extend the previous findings to provide evidence that mirror visual feedback engages the functional networks associated with the perceptual–attentional process and triggers M1 activation, although the M1 activation is functionally independent of other brain regions unrelated to motor function. In summary, this study demonstrated a concrete functional connectivity pattern for motor control in the face of visual conflicts, and providing a foundation for future research to examine the dynamic functional networks of mirror illusion in motor control.

Treatment of triple-negative breast cancer (TNBC) remains an unmet clinical need owing to its lack of an efficient therapeutic target. The targeting of DNA repair by poly(ADP-ribose) polymerase (PARP) inhibitors has shown benefit for patients with the BRCA variation. However, sensitivities to the PARP inhibitors were reported regardless of BRCA status. Thus, exploring the underlying mechanisms is imperative. Herein, we identified that breast cancer cells with an elevated expression of protein arginine methyl transferase 1 (PRMT1) was associated with therapeutic sensitivity to the PARP inhibitor olaparib. The results of cell viability and colony formation assays indicated that the suppression of PRMT1 by small hairpin RNA or by the chemical inhibitor increased sensitivity to olaparib in human TNBC MDA-MB-231 and BT549 cells. Bioinformatic analysis revealed that PRMT1 expression was significantly associated with the MYC signature, and TNBC cells with higher PRMT1 and the MYC signature were associated with therapeutic sensitivity to olaparib. Mechanistic studies further demonstrated that knockdown of PRMT1 reduced the c-Myc protein level and downregulated the expression of MYC downstream targets, whereas overexpression of PRMT1 enhanced c-Myc protein expression. Moreover, the overexpression of PRMT1 promoted c-Myc protein stability, and the inhibition of PRMT1 downregulated c-Myc protein stability. Accordingly, the knockdown of PRMT1 inhibited homologous recombination gene expression. These data indicate that PRMT1 is instrumental in regulating DNA repair, at least in part, by modulating c-Myc signaling. Our data highlighted the PRMT1/c-Myc network as a potential therapeutic target in patients with TNBC.

Background. Several brain regions are activated in response to mirror visual feedback (MVF). However, less is known about how these brain areas and their connectivity are modulated in stroke patients. This study aimed to explore the effects of MVF on brain functional connectivity in stroke patients. Materials and Methods. We enrolled 15 stroke patients who executed Bilateral-No mirror, Bilateral-Mirror, and Unilateral-Mirror conditions. The coherence values among five brain regions of interest in four different frequency bands were calculated from magnetoencephalographic signals. We examined the differences in functional connectivity of each two brain areas between the Bilateral-No mirror and Bilateral-Mirror conditions and between the Bilateral-Mirror and Unilateral-Mirror conditions. Results. The functional connectivity analyses revealed significantly stronger connectivity between the posterior cingulate cortex and primary motor cortex in the beta band (adjusted p = 0.04) and possibly stronger connectivity between the precuneus and primary visual cortex in the theta band (adjusted p = 0.08) in the Bilateral-Mirror condition than those in the Bilateral-No mirror condition. However, the comparisons between the Bilateral-Mirror and Unilateral-Mirror conditions revealed no significant differences in cortical coherence in all frequency bands. Conclusions. Providing MVF to stroke patients may modulate the lesioned primary motor cortex through visuospatial and attentional cortical networks.

Abstract Background Loss of upper-extremity motor function is one of the most debilitating deficits following stroke. Two promising treatment approaches, action observation therapy (AOT) and mirror therapy (MT), aim to enhance motor learning and promote neural reorganization in patients through different afferent inputs and patterns of visual feedback. Both approaches involve different patterns of motor observation, imitation, and execution but share some similar neural bases of the mirror neuron system. AOT and MT used in stroke rehabilitation may confer differential benefits and neural activities that remain to be determined. This clinical trial aims to investigate and compare treatment effects and neural activity changes of AOT and MT with those of the control intervention in patients with subacute stroke. Methods/design An estimated total of 90 patients with subacute stroke will be recruited for this study. All participants will be randomly assigned to receive AOT, MT, or control intervention for a 3-week training period (15 sessions). Outcome measurements will be taken at baseline, immediately after treatment, and at the 3-month follow-up. For the magnetoencephalography (MEG) study, we anticipate that we will recruit 12 to 15 patients per group. The primary outcome will be the Fugl-Meyer Assessment score. Secondary outcomes will include the modified Rankin Scale, the Box and Block Test, the ABILHAND questionnaire, the Questionnaire Upon Mental Imagery, the Functional Independence Measure, activity monitors, the Stroke Impact Scale version 3.0, and MEG signals. Discussion This clinical trial will provide scientific evidence of treatment effects on motor, functional outcomes, and neural activity mechanisms after AOT and MT in patients with subacute stroke. Further application and use of AOT and MT may include telerehabilitation or home-based rehabilitation through web-based or video teaching. Trial registration ClinicalTrials.gov, ID: NCT02871700 . Registered on 1 August 2016.

Mirror visual feedback (MVF) has been shown to increase the excitability of the primary motor cortex (M1) during asynchronous bimanual movement. However, the functional networks underlying this process remain unclear. We recruited 16 healthy volunteers to perform asynchronous bimanual movement, that is, their left hand performed partial range of movement while their right hand performed normal full range of movement. Their ongoing brain activities were recorded by whole-head magnetoencephalography during the movement. Participants were required to keep both hands stationary in the control condition. In the other two conditions, participants were required to perform asynchronous bimanual movement with MVF (Asy_M) and without MVF (Asy_w/oM). Greater M1 excitability was found under Asy_M than under Asy_w/oM. More importantly, when receiving MVF, the visual cortex reduced its functional connection to brain regions associated with perceptuo-motor-attentional process (i.e., M1, superior temporal gyrus, and dorsolateral prefrontal cortex). This is the first study to demonstrate a global functional network of MVF during asynchronous bimanual movement, providing a foundation for future research to examine the neural mechanisms of mirror illusion in motor control.

Abstract While the automatic inhibitory function of the human cerebral cortex has been extensively investigated by means of electrophysiological recordings, the corresponding modulating neurochemical mechanisms remain unclear. We aimed to examine whether the primary somatosensory (SI) and primary motor cortical (MI) inhibitory function is associated with endogenous GABA levels. Eighteen young participants received paired-pulse and single-pulse electrical stimulation to the median nerve during magnetoencephalographic recordings. The SI sensory gating (SG), considered as an automatic inhibitory ability, was measured as the amplitude ratio of Stimulus 2 over Stimulus 1, in the paired-pulse paradigm. In addition, stimulus-induced beta activity, considered to originate from MI and also to be related to inhibitory function, was estimated using the single-pulse paradigm. The GABA+ concentration of the sensorimotor cortex was acquired from each subject by using magnetic resonance spectroscopy (MRS). A lower SG ratio in SI was significantly associated with an increased beta power in MI. More importantly, the beta rebound power, but not SI SG ratio, was positively correlated with GABA+ concentration. Our findings show a tight functional relationship between SI and MI during processing of automatic inhibition. GABA+ levels appear to be more closely related to the automatic inhibitory function of MI than SI.

Abstract The insulin-like growth factor (IGF)-1 signaling is relevant in regulating cell growth and cytokine secretions by glioblastomas. MicroRNAs determine the cell fate in glioblastomas. However, relationships between IGF-1 signaling and miRNAs in glioblastoma pathogenesis are still unclear. Our aim was to validate the IGF-1-mediated mRNA/miRNA regulatory network in glioblastomas. Using in silico analyses of mRNA array and RNA sequencing data from The Cancer Genome Atlas (TCGA), we identified 32 core enrichment genes that were highly associated with IGF-1-promoted cytokine-cytokine receptor interactions. To investigate the IGF-1-downregulated miRNA signature, microarray-based approaches with IGF-1-treated U87-MG cells and array data in TCGA were used. Four miRNAs, including microRNA (miR)-9-5p, miR-9-3p, miR-181d, and miR-130b, exhibited an inverse correlation with IGF-1 levels. The miR-181d, that targeted the most IGF-1-related cytokine genes, was significantly reduced in IGF-1-treated glioma cells. Statistical models incorporating both high-IGF-1 and low-miR-181d statuses better predicted poor patient survival, and can be used as an independent prognostic factor in glioblastomas. The C-C chemokine receptor type 1 (CCR1) and interleukin (IL)-1b demonstrated inverse correlations with miR-181d levels and associations with patient survival. miR-181d significantly attenuated IGF-1-upregulated CCR1 and IL-1b gene expressions. These findings demonstrate a distinct role for IGF-1 signaling in glioma progression via miR-181d/cytokine networks.

Temozolomide (TMZ) is a first-line alkylating agent for glioblastoma multiforme (GBM). Clarifying the mechanisms inducing TMZ insensitivity may be helpful in improving its therapeutic effectiveness against GBM. Insulin-like growth factor (IGF)-1 signaling and micro (mi)RNAs are relevant in mediating GBM progression. However, their roles in desensitizing GBM cells to TMZ are still unclear. We aimed to identify IGF-1-mediated miRNA regulatory networks that elicit TMZ insensitivity for GBM. IGF-1 treatment attenuated TMZ cytotoxicity via WNT/β-catenin signaling, but did not influence glioma cell growth. By miRNA array analyses, 93 upregulated and 148 downregulated miRNAs were identified in IGF-1-treated glioma cells. miR-513a-5p from the miR-513a-2 gene locus was upregulated by IGF-1-mediated phosphoinositide 3-kinase (PI3K) signaling. Its elevated levels were also observed in gliomas versus normal cells, in array data of The Cancer Genome Atlas (TCGA), and the GSE61710, GSE37366, and GSE41032 datasets. In addition, lower levels of neural precursor cell-expressed developmentally downregulated 4-like (NEDD4L), an E3 ubiquitin protein ligase that inhibits WNT signaling, were found in gliomas by analyzing cells, arrays, and RNA sequencing data of TCGA glioma patients. Furthermore, a negative correlation was identified between miR-513a-5p and NEDD4L in glioma. NEDD4L was also validated as a direct target gene of miR-513a-5p, and it was reduced by IGF-1 treatment. Overexpression of NEDD4L inhibited glioma cell viability and reversed IGF-1-repressed TMZ cytotoxicity. In contrast, miR-513a-5p significantly affected NEDD4L-inhibited WNT signaling and reduced TMZ cytotoxicity. These findings demonstrate a distinct role of IGF-1 signaling through miR-513a-5p-inhibited NEDD4L networks in influencing GBM's drug sensitivity to TMZ.

Action observation therapy has recently attracted increasing attention; however, the mechanisms through which action observation and execution (AOE) modulate neural activity in stroke patients remain unclear. This study was aimed at investigating the effects of action observation and two types of AOE on motor cortical activations after stroke using magnetoencephalography. Twenty patients with stroke and 20 healthy controls were recruited for the collection of data on the beta oscillatory activity in the primary motor cortex (M1). All participants performed the conditions of resting, observation only, and video observation combined with execution (video AOE). Stroke patients performed one additional condition of affected hand observation combined with execution (affected hand AOE). The relative change index of beta oscillations was calculated, and nonparametric tests were used to examine the differences in conditions. In stroke patients, the relative change index of M1 beta oscillatory activity under the video AOE condition was significantly lower than that under the observation only and affected hand AOE conditions. Moreover, M1 cortical activity did not significantly differ under the observation only and affected hand AOE conditions. For healthy controls, the relative change index under the video AOE condition was significantly lower than that under the observation only condition. In addition, no significant differences in relative change indices were found under the observation only and video AOE conditions between the 2 groups. This study provides new insight into the neural mechanisms underlying AOE, which supports the use of observing videos of normal movements during action observation therapy in stroke rehabilitation.

Inhibition, the ability to suppress irrelevant information, thoughts or movements, is crucial for humans to perform context-appropriate behaviors. It was suggested that declined cognitive performance in older adults might be attributed to inhibitory deficiencies. Although previous studies have shown an age-associated reduction in inhibitory ability, the understanding regarding its cortical spatiotemporal maps remained limited. Thus, we used a whole-head magnetoencephalography (MEG) to elucidate the age effects on response inhibition, and to explore the brain activation differences in high- and low-performing seniors. We recruited 22 younger and 22 older adults to participate in the visual Go/No-go task. Both behavioral performance and neuromagnetic responses to No-go stimuli were analyzed. The behavioral results showed that the older adults made more false alarm (FA) errors than the younger adults did. The MEG results showed that the seniors exhibited declined cortical activities in middle temporal gyrus (MTG) and delayed activation in MTG, prefrontal cortex (PFC) and pre-supplementary motor area (pre-SMA). Furthermore, among the older adults, more recruitment of the left PFC was found in the high-performers than in the lower-performers. In conclusion, age-related deficiencies in response inhibition were observed in both behavioral performance and neurophysiological measurement. Our results also suggested that frontal recruitment plays a compensatory role in successful inhibition.

Response inhibition is frequently examined using visual go/no-go tasks. Recently, the auditory go/no-go paradigm has been also applied to several clinical and aging populations. However, age-related changes in the neural underpinnings of auditory go/no-go tasks are yet to be elucidated. We used magnetoencephalography combined with distributed source imaging methods to examine age-associated changes in neural responses to auditory no-go stimuli. Additionally, we compared the performance of high- and low-performing older adults to explore differences in cortical activation. Behavioral performance in terms of response inhibition was similar in younger and older adult groups. Relative to the younger adults, the older adults exhibited reduced cortical activation in the superior and middle temporal gyrus. However, we did not find any significant differences in cortical activation between the high- and low-performing older adults. Our results therefore support the hypothesis that inhibition is reduced during aging. The variation in cognitive performance among older adults confirms the need for further study on the underlying mechanisms of inhibition.

The dietary flavonoids luteolin and quercetin are reported to inhibit cancer mobility; however, the regulatory effect of luteolin and quercetin on RPS12 is still unclear. Here, we found that A431-III cells expressed a higher level of RPS12 than A431-P cells. The flavonoids luteolin and quercetin reduced RPS12 and c-Myc expressions via Akt/mTOR signalling. The Akt inhibitor LY294002 and mTOR inhibitor rapamycin reduced RPS12 and c-Myc expressions. The c-Myc inhibitor 10058-F4 reduced RPS12 expression and promoter transactivation. The overexpression of c-Myc increased RPS12 expression. Akt, mTOR, and c-Myc inhibitor blocked cell migration. Reducing RPS12 expression via 10058-F4 and shRNAs reduced cell invasion. This study reveals that RPS12 is upregulated via Akt/mTOR/c-Myc signalling and increased cell mobility. Luteolin and quercetin blocked Akt/mTOR/c-Myc signalling to reduce RPS12 level and downstream cell mobility. These data suggest a possible role of RPS12 in cell mobility and may be a potential therapy target for cervical cancer.

Human high-mobility group A2 (HMGA2) encodes for a non-histone chromatin protein which influences a variety of biological processes, including the cell cycle process, apoptosis, the DNA damage repair process, and epithelial−mesenchymal transition. The accumulated evidence suggests that high expression of HMGA2 is related to tumor progression, poor prognosis, and a poor response to therapy. Thus, HMGA2 is an important molecular target for many types of malignancies. Our recent studies revealed the positive connections between heat shock protein 90 (Hsp90) and HMGA2 and that the Hsp90 inhibitor has therapeutic potential to inhibit HMGA2-triggered tumorigenesis. However, 43% of patients suffered visual disturbances in a phase I trial of the second-generation Hsp90 inhibitor, NVP-AUY922. To identify a specific inhibitor to target HMGA2, the Gene Expression Omnibus (GEO) database and the Library of Integrated Network-based Cellular Signatures (LINCS) L1000platform were both analyzed. We identified the approved small-molecule antifungal agent ciclopirox (CPX) as a novel potential inhibitor of HMGA2. In addition, CPX induces cytotoxicity of colorectal cancer (CRC) cells by induction of cell cycle arrest and apoptosis in vitro and in vivo through direct interaction with the AT-hook motif (a small DNA-binding protein motif) of HMGA2. In conclusion, this study is the first to report that CPX is a novel potential inhibitor of HMGA2 using a drug-repurposing approach, which can provide a potential therapeutic intervention in CRC patients.

Flavonoids are well-known antioxidants and have shown the ability to prevent tumor formation and recurrence. Especially in dietary flavonoids, they have provided convenience and consistence of intake for long-term prevention of tumor formation. Previous reports suggested that S100 calcium-binding protein A7 (S100A7) might activate epithelial−mesenchymal transition (EMT) signaling and promote the metastasis of tumor cells; however, the regulatory signaling was unclear. In this study, we found that S100A7 was highly expressed in cancer cells and could be reduced by luteolin (Lu) and quercetin (Qu) through Src/Stat3 signaling. We found that the protein levels of S100A7, phosphorylated Src (p-Src), and p-Stat3 were increased in A431-III cells. Flavonoids Lu and Qu reduce protein levels of p-Src, p-Stat3 and S100A7 in A431-III cells. Treatment of A431-III cells with Src inhibitor SU6656 and Stat3 inhibitor S3I-201 also reduced the protein levels of S100A7. Transactivation activity of 5′-upstream regions of S100A7 was activated by Stat3 but was reduced by treatment with Lu, Qu, SU6656 and S3I-201. The treatment also reduced the migratory and invasive abilities of A431-III cells. In a further analysis of EMT markers, the protein level of E-cad increased and that of Twist decreased after treatment with the inhibitors and flavonoids. Overexpression of S100A7 decreased the protein level of E-cad and increased the Twist level, whereas knockdown of S100A7 had the opposite effects. Treatment with S3I-201, Lu and Qu, compared to the control, were found to decrease metastasis of tumor cells in zebrafish larvae. These results suggest that Lu and Qu may inhibit Src/Stat3/S100A7 signaling to reduce tumorigenesis of cancer cells.

Age-related deficiency in the top-down modulation of cognitive inhibition has been extensively documented, whereas the effects of age on a bottom-up or automatic operation of inhibitory function were less investigated. It is unknown that whether the older adults (OA)’ reduced behavioral performance and neural responses are due to the insufficient bottom-up processes. Compared to behavioral assessments which have been widely used to examine the top-down control of response inhibition, electrophysiological recordings are more suitable to probe the early-stage processes of automatic inhibitory function. Sensory gating (SG), a phenomenon of attenuated neural response to the second identical stimulus in a paired-pulse paradigm, is an indicator to assess automatic inhibitory function of the sensory cortex. On the other hand, electricity-induced beta rebound oscillation in a single-pulse paradigm reflects cortical inhibition of the motor cortex. From the neurophysiological perspective, SG and beta rebound oscillation are replicable indicators to examine the automatic inhibitory function of human sensorimotor cortices. Thus, the present study aimed to use a whole-head magnetoencephalography (MEG) to investigate the age-related alterations of SG function in the primary somatosensory cortex (SI) and of beta rebound oscillation in the primary motor cortex (MI) in 17 healthy younger and 15 older adults. The Stimulus 2/Stimulus 1 (S2/S1) amplitude ratio in response to the paired-pulse electrical stimulation to the left median nerve was used to evaluate the automatic inhibitory function of SI, and the beta rebound response in the single-pulse paradigm was used to evaluate the automatic inhibitory function of MI. Although there were no significant age-related differences found in the SI SG ratios, the MI beta rebound power was reduced and peak latency was prolonged in the OA. Furthermore, significant association between the SI SG ratio and the MI beta rebound power, which was seen in the younger adults (YA), was absent in the OA. In conclusion, our data suggested an age-related defect of association between sensorimotor cortices regarding automatic inhibitory function.

MicroRNAs are small noncoding RNAs that post-transcriptionally control the expression of genes involved in glioblastoma multiforme (GBM) development. Although miR-302b functions as a tumor suppressor, its role in GBM is still unclear. Therefore, this study comprehensively explored the roles of miR-302b-mediated gene networks in GBM cell death. We found that miR-302b levels were significantly higher in primary astrocytes than in GBM cell lines. miR-302b overexpression dose dependently reduced U87-MG cell viability and induced apoptosis through caspase-3 activation and poly(ADP ribose) polymerase degradation. A transcriptome microarray revealed 150 downregulated genes and 380 upregulated genes in miR-302b-overexpressing cells. Nuclear factor IA (NFIA), higher levels of which were significantly related to poor survival, was identified as a direct target gene of miR-302b and was involved in miR-302b-induced glioma cell death. Higher NFIA levels were observed in GBM cell lines and human tumor sections compared with astrocytes and non-tumor tissues, respectively. NFIA knockdown significantly enhanced apoptosis. We found high levels of insulin-like growth factor-binding protein 2 (IGFBP2), another miR-302b-downregulated gene, in patients with poor survival. We verified that NFIA binds to the IGFBP2 promoter and transcriptionally enhances IGFBP2 expression levels. We identified that NFIA-mediated IGFBP2 signaling pathways are involved in miR-302b-induced glioma cell death. The identification of a regulatory loop whereby miR-302b inhibits NFIA, leading to a decrease in expression of IGFBP-2, may provide novel directions for developing therapies to target glioblastoma tumorigenesis.

Colorectal cancer (CRC) is a heterogeneous disease resulting from the combined influence of many genetic factors. This complexity has caused the molecular characterization of CRC to remain uncharacterized, with a lack of clear gene markers associated with CRC and the prognosis of this disease. Thus, highly sensitive tumor markers for the detection of CRC are the most essential determinants of survival. In this study, we examined the simultaneous downregulation of the mRNA levels of six metallothionein (MT) genes in CRC cell lines and public CRC datasets for the first time. In addition, we detected downregulation of these six MT mRNAs’ levels in 30 pairs of tumor (T) and adjacent non-tumor (N) CRC specimens. In order to understand the potential prognostic relevance of these six MT genes and CRC, we presented a four-gene signature to evaluate the prognosis of CRC patients. Further discovery suggested that the four-gene signature (MT1F, MT1G, MT1L, and MT1X) predicted survival better than any combination of two-, three-, four-, five-, or six-gene models. In conclusion, this study is the first to report that simultaneous downregulation of six MT mRNAs’ levels in CRC patients, and their aberrant expression together, accurately predicted CRC patients’ outcomes.

The rhizoma of Ligusticum sinense, a Chinese medicinal plant, has long been used as a cosmetic for the whitening and hydrating of the skin in ancient China. In order to investigate the antimelanogenic components of the rhizoma of L. sinense, we performed an antimelanogenesis assay-guided purification using semi-preparative HPLC accompanied with spectroscopic analysis to determine the active components. Based on the bioassay-guided method, 24 compounds were isolated and identified from the ethyl acetate layer of methanolic extracts of L. sinense, and among these, 5-[3-(4-hydroxy-3-methoxyphenyl)allyl]ferulic acid (1) and cis-4-pentylcyclohex-3-ene-1,2-diol (2) were new compounds. All the pure isolates were subjected to antimelanogenesis assay using murine melanoma B16-F10 cells. Compound 1 and (3S,3aR)-neocnidilide (8) exhibited antimelanogenesis activities with IC50 values of 78.9 and 31.1 μM, respectively, without obvious cytotoxicity. Further investigation showed that compound 8 demonstrated significant anti-pigmentation activity on zebrafish embryos (10‒20 μM) compared to arbutin (20 μM), and without any cytotoxicity against normal human epidermal keratinocytes. These findings suggest that (3S,3aR)-neocnidilide (8) is a potent antimelanogenic and non-cytotoxic natural compound and may be developed potentially as a skin-whitening agent for cosmetic uses.

Temozolomide (TMZ), an alkylating agent of the imidazotetrazine series, is a first-line chemotherapeutic drug used in the clinical therapy of glioblastoma multiforme, the most common and high-grade primary glioma in adults. Micro (mi)RNAs, which are small noncoding RNAs, post-transcriptionally regulate gene expressions and are involved in gliomagenesis. However, no studies have reported relationships between TMZ and miRNA gene regulation. We investigated TMZ-mediated miRNA profiles and its molecular mechanisms underlying the induction of glioma cell death. By performing miRNA microarray and bioinformatics analyses, we observed that expression of 248 miRNAs was altered, including five significantly upregulated and 17 significantly downregulated miRNAs, in TMZ-treated U87MG cells. miR-128 expression levels were lower in different glioma cells and strongly associated with poor survival. TMZ treatment significantly upregulated miR-128 expression. TMZ significantly enhanced miR-128-1 promoter activity and transcriptionally regulated miR-128 levels through c-Jun N-terminal kinase 2/c-Jun pathways. The overexpression and knockdown of miR-128 expression significantly affected TMZ-mediated cell viability and apoptosis-related protein expression. Furthermore, the overexpression of miR-128 alone enhanced apoptotic death of glioma cells through caspase-3/9 activation, poly(ADP ribose) polymerase degradation, reactive oxygen species generation, mitochondrial membrane potential loss, and non-protective autophagy formation. Finally, we identified that key members in mammalian target of rapamycin (mTOR) signaling including mTOR, rapamycin-insensitive companion of mTOR, insulin-like growth factor 1, and PIK3R1, but not PDK1, were direct target genes of miR-128. TMZ inhibited mTOR signaling through miR-128 regulation. These results indicate that miR-128-inhibited mTOR signaling is involved in TMZ-mediated cytotoxicity. Our findings may provide a better understanding of cytotoxic mechanisms of TMZ involved in glioblastoma development.

Colon cancer is the third most common cancer in the world and the second most common cause of cancer-related mortality. Molecular biomarkers for colon cancer have undergone vigorous discovery and validation. Recent studies reported that overexpression of podocalyxin-like protein 1 (PODXL) is associated with distant metastasis and poor prognosis across several types of malignancies. Its role and underlying molecular mechanism, however, are not yet fully understood. In the present study, we revealed that the Hippo transducer, the transcriptional coactivator with PDZ-binding motif (TAZ), acts as a downstream mediator of PODXL in colon cancer. Inhibition of PODXL resulted in the suppression of TAZ signaling and the downregulation of Hippo downstream genes. Moreover, PODXL plays a critical role in cancer stemness, invasiveness, and sensitivity to chemotherapies in colon cancer HCT15 cells. Notably, expression of PODXL showed a positive correlation with stem-like and epithelial-mesenchymal transition (EMT) core signatures, and was associated with poor survival outcomes in patients with colon cancer. These findings provide novel insights into the molecular mechanism of PODXL-mediated tumorigenesis in colon cancer.

Sensory gating (SG), referring to an attenuated neural response to the second identical stimulus, is considered as preattentive processing in the central nervous system to filter redundant sensory inputs. Insufficient somatosensory SG has been found in the aged adults, particularly in the secondary somatosensory cortex (SII). However, it remains unclear which variables leading to the age-related somatosensory SG decline. There has been evidence showing a relationship between brain oscillations and cortical evoked excitability. Thus, this study used whole-head magnetoencephalography to record responses to paired-pulse electrical stimulation to the left median nerve in healthy young and elderly participants to test whether insufficient stimulus 1- (S1-) induced event-related desynchronization (ERD) contributes to a less-suppressed stimulus 2- (S2-) evoked response. Our analysis revealed that the minimum norm estimates showed age-related reduction of SG in the bilateral SII regions. Spectral power analysis showed that the elderly demonstrated significantly reduced alpha ERD in the contralateral SII (SIIc). Moreover, it was striking to note that lower S1-induced alpha ERD was associated with higher S2-evoked amplitudes in the SIIc among the aged adults. Conclusively, our findings suggest that age-related decline of somatosensory SG is partially attributed to the altered S1-induced oscillatory activity.

The perception of respiratory sensations can be of significant importance to individuals for survival and greatly impact quality of life. Respiratory sensory gating, similar to somatosensory gating with exteroceptive stimuli, is indicative of brain cortices filtering out repetitive respiratory stimuli and has been investigated in adults with and without diseases. Respiratory gating can be tested with the respiratory-related evoked potential (RREP) method in the electroencephalogram with a paired inspiratory occlusion paradigm. Here, the RREP N1 component elicited by the second stimulus (S2) shows reduced amplitudes compared to the RREP N1 component elicited by the first stimulus (S1). However, little is known regarding the effect of development on respiratory sensory gating. The present study examined respiratory sensory gating in 22 typically developed school-aged children and 22 healthy adults. Paired inspiratory occlusions of 150-ms each with an inter-stimulus-interval of 500-ms were delivered randomly every 2–4 breaths during recording. The main results showed a significantly larger RREP N1 S2/S1 ratio in the children group than in the adult group. In addition, children compared to adults demonstrated significantly smaller N1 peak amplitudes in response to S1. Our results suggest that school-aged children, compared to adults, display reduced respiratory sensory gating.

The fish lateral line (LL) is a mechanosensory system closely related to the hearing system of higher vertebrates, and it is composed of several neuromasts located on the surface of the fish. These neuromasts can detect changes in external water flow, to assist fish in maintaining a stationary position in a stream. In the present study, we identified a novel function of Nogo/Nogo receptor signaling in the formation of zebrafish neuromasts. Nogo signaling in zebrafish, like that in mammals, involves three ligands and four receptors, as well as three co-receptors (TROY, p75, and LINGO-1). We first demonstrated that Nogo-C2, NgRH1a, p75, and TROY are able to form a Nogo-C2 complex, and that disintegration of this complex causes defective neuromast formation in zebrafish. Time-lapse recording of the CldnB::lynEGFP transgenic line revealed that functional obstruction of the Nogo-C2 complex causes disordered morphogenesis, and reduces rosette formation in the posterior LL (PLL) primordium during migration. Consistent with these findings, hair-cell progenitors were lost from the PLL primordium in p75, TROY, and Nogo-C2/NgRH1a morphants. Notably, the expression levels of pea3, a downstream marker of Fgf signaling, and dkk1b, a Wnt signaling inhibitor, were both decreased in p75, TROY, and Nogo-C2/NgRH1a morphants; moreover, dkk1b mRNA injection could rescue the defects in neuromast formation resulting from knockdown of p75 or TROY. We thus suggest that a novel Nogo-C2 complex, consisting of Nogo-C2, NgRH1a, p75, and TROY, regulates Fgf signaling and dkk1b expression, thereby ensuring stable organization of the PLL primordium.

Metastasis is a major cause of mortality in cancer patients. Invadopodia are considered to be crucial structures that allow cancer cells to penetrate across the extracellular matrix (ECM) by using matrix metalloproteinases (MMPs). Previously, we isolated a highly invasive A431-III subline from parental A431 cells by Boyden chamber assay. The A431-III cells possess higher invasive and migratory abilities, elevated levels of MMP-9 and an enhanced epithelial-mesenchymal transition (EMT) phenotype. In this study, we discovered that A431-III cells had an increased potential to form invadopodia and an improved capacity to degrade ECM compared with the original A431 cells. We also observed enhanced phosphorylation levels of cortactin and Src in A431-III cells; these phosphorylated proteins have been reported to be the main regulators of invadopodia formation. Flavonoids, almost ubiquitously distributed in food plants and plant food products, have been documented to exhibit anti-tumor properties. Therefore, it was of much interest to explore the effects of flavonoid antioxidants on the metastatic activity of A431-III cells. Exposure of A431-III cells to two potent dietary flavonoids, namely luteolin (Lu) and quercetin (Qu), caused inhibition of invadopodia formation and decrement in ECM degradation. We conclude that Lu and Qu attenuate the phosphorylation of cortactin and Src in A431-III cells. As a consequence, there ensues a disruption of invadopodia generation and the suppression of MMP secretion. These changes, in concert, bring about a reduction in metastasis.

BackgroundThe zona pellucida (ZP) domain is part of many extracellular proteins with diverse functions from structural components to receptors. The mammalian β-tectorin is a protein of 336 amino acid residues containing a single ZP domain and a putative signal peptide at the N-terminus of the protein. It is 1 component of a gel-like structure called the tectorial membrane which is involved in transforming sound waves into neuronal signals and is important for normal auditory function. β-Tectorin is specifically expressed in the mammalian and avian inner ear.Methodology/principal findingsWe identified and cloned the gene encoding zebrafish β-tectorin. Through whole-mount in situ hybridization, we demonstrated that β-tectorin messenger RNA was expressed in the otic placode and specialized sensory patch of the inner ear during zebrafish embryonic stages. Morpholino knockdown of zebrafish β-tectorin affected the position and number of otoliths in the ears of morphants. Finally, swimming behaviors of β-tectorin morphants were abnormal since the development of the inner ear was compromised.Conclusions/significanceOur results reveal that zebrafish β-tectorin is specifically expressed in the zebrafish inner ear, and is important for regulating the development of the zebrafish inner ear. Lack of zebrafish β-tectorin caused severe defects in inner ear formation of otoliths and function.

BACKGROUND:Mammalian M6A, a member of the proteolipid protein (PLP/DM20) family expressed in neurons, was first isolated by expression cloning with a monoclonal antibody. Overexpression of M6A was shown to induce filopodium formation in neuronal cells; however, the underlying mechanism of is largely unknown. Possibly due to gene duplication, there are two M6A paralogs, M6Aa and M6Ab, in the zebrafish genome. In the present study, we used the zebrafish as a model system to investigate the role of zebrafish M6Ab in filopodium formation in PC12 cells and neurite outgrowth in zebrafish embryos. METHODOLOGY/PRINCIPAL FINDINGS:We demonstrated that zebrafish M6Ab promoted extensive filopodium formation in NGF-treated PC12 cells, which is similar to the function of mammalian M6A. Phosphorylation at serine 263 of zebrafish M6Ab contributed to this induction. Transfection of the S263A mutant protein greatly reduced filopodium formation in PC12 cells. In zebrafish embryos, only S263D could induce neurite outgrowth. CONCLUSIONS/SIGNIFICANCE:Our results reveal that the phosphorylation status of zebrafish M6Ab at serine 263 is critical for its role in regulating filopodium formation and neurite outgrowth.