Investigation of SIS epidemics on dynamic network models with temporary link deactivation control schemes.

Mathematical modeling of epidemic diseases is increasingly being used to respond to emerging diseases. Although conditions modeled by SIS dynamics will eventually reach either a disease-free steady-state or an endemic steady state without interventions, it is desirable to eradicate the disease as quickly as possible by introducing a control scheme. Here, we investigate the control methods of epidemic models on dynamic networks with temporary link deactivation. A quick link deactivation mechanism can simulate a community effort to reduce the risk of infection by temporarily avoiding infected neighbors. Once infected individuals recover, the links between the susceptible and recovered are activated. Our study suggests that a control scheme that has been shown ineffective in controlling dynamic network models may yield effective responses for networks with certain types of link dynamics, such as the temporary link deactivation mechanisms. We observe that a faster and more effective eradication could be achieved by updating control schemes frequently.

16p12.1 Deletion Orthologs are Expressed in Motile Neural Crest Cells and are Important for Regulating Craniofacial Development in Xenopus laevis.

Copy number variants (CNVs) associated with neurodevelopmental disorders are characterized by extensive phenotypic heterogeneity. In particular, one CNV was identified in a subset of children clinically diagnosed with intellectual disabilities (ID) that results in a hemizygous deletion of multiple genes at chromosome 16p12.1. In addition to ID, individuals with this deletion display a variety of symptoms including microcephaly, seizures, cardiac defects, and growth retardation. Moreover, patients also manifest severe craniofacial abnormalities, such as micrognathia, cartilage malformation of the ears and nose, and facial asymmetries; however, the function of the genes within the 16p12.1 region have not been studied in the context of vertebrate craniofacial development. The craniofacial tissues affected in patients with this deletion all derive from the same embryonic precursor, the cranial neural crest, leading to the hypothesis that one or more of the 16p12.1 genes may be involved in regulating neural crest cell (NCC)-related processes. To examine this, we characterized the developmental role of the 16p12.1-affected gene orthologs, polr3e, mosmo, uqcrc2, and cdr2, during craniofacial morphogenesis in the vertebrate model system, Xenopus laevis. While the currently-known cellular functions of these genes are diverse, we find that they share similar expression patterns along the neural tube, pharyngeal arches, and later craniofacial structures. As these genes show co-expression in the pharyngeal arches where NCCs reside, we sought to elucidate the effect of individual gene depletion on craniofacial development and NCC migration. We find that reduction of several 16p12.1 genes significantly disrupts craniofacial and cartilage formation, pharyngeal arch migration, as well as NCC specification and motility. Thus, we have determined that some of these genes play an essential role during vertebrate craniofacial patterning by regulating specific processes during NCC development, which may be an underlying mechanism contributing to the craniofacial defects associated with the 16p12.1 deletion.

Tau interferes with axonal neurite stabilization and cytoskeletal composition independently of its ability to associate with microtubules.

Tau impacts overall axonal transport particularly when overexpressed by interfering with translocation of kinesin along microtubules (MTs) and/or as a cargo of kinesin by outcompeting other kinesin cargo. To discern between which of these mechanisms was more robust during axonal outgrowth, we overexpressed phosphomimetic (E18; which is incapable of MT binding), phospho-null (A18) or wild-type (WT) full-length human tau conjugated to EGFP, the latter two of which bind MTs. Expression of WT and A18 displayed increased acetylated MTs and resistance to colchicine, while expression of E18 did not, indicating that E18 did not contribute to MT stabilization. Expression of all tau constructs reduced overall levels of neurofilaments (NFs) within axonal neurites, and distribution of NFs along neurite lengths. Since NFs are another prominent cargo of kinesin during axonal neurite outgrowth, this finding is consistent with WT, A18 and E18 inhibiting NF transport to the same extent by competing as cargo of kinesin. These findings indicate that tau can impair axonal transport independently of association with MTs in growing axonal neurites.

Corrigendum: Wolf-Hirschhorn Syndrome-Associated Genes Are Enriched in Motile Neural Crest Cells and Affect Craniofacial Development in Xenopus laevis.

[This corrects the article DOI: 10.3389/fphys.2019.00431.].

The Influence of Tourists' Experience of Quality of Street Foods on Destination's Image, Life Satisfaction, and Word of Mouth: The Moderating Impact of Food Neophobia.

Due to growing food-related tourism, there is increasing interest about street foods worldwide, including South Korea. Many types of food-related experiences have been considered as one of the significant elements to develop positive perceptions about a destination, and street food has been recognized as a critical clue for encouraging tourists to a destination. Previous scholars mentioned street food as a public health risk element as well as a significant factor to attract tourists' attention. Therefore, this study aims to find out how experiential quality of street foods is related to the destination image, life satisfaction, and word of mouth as perceived by tourists in night markets of South Korea. Data was collected from 325 foreigners who visited night markets and have experienced street foods in Korea. This study demonstrates the results of the influence of quality of street foods on tourist experience, on destination image, on life satisfaction, and on word of mouth in Korea. In addition, the result shows a moderating impact of food neophobia on the formulated relationships. There are statistically significant differences between groups with high neophobia perception and low neophobia perception of street foods. Based on the results of this study, we propose not only academic implications for future studies, but also managerial implications for food enterprises and food tourism organizers related to street food.

Neurofilaments form flexible bundles during neuritogenesis in culture and in mature axons in situ.

Neurofilaments (NFs) undergo cation-dependent phospho-mediated associations with each other and other cytoskeletal elements that support axonal outgrowth. Progressive NF-NF associations generate a resident, bundled population that undergoes exchange with transporting NFs. We examined the properties of bundled NFs. Bundles did not always display a fully linear profile but curved and twisted at various points along the neurite length. Bundles retracted faster than neurites and retracted bundles did not expand following extraction with Triton, indicating that they coiled passively rather than due to pressure from the cell. Bundles consisted of helically wound NFs, which may provide flexibility necessary for turning of growing axons during pathfinding. Interactions between NFs and other cytoskeletal elements may be disrupted en masse during neurite retraction or regionally during remodeling. It is suggested that bundles within long axons that cannot be fully retracted into the soma could provide maintain proximal support yet still allow more distal flexibility for remodeling and changing direction during pathfinding.

Wolf-Hirschhorn Syndrome-Associated Genes Are Enriched in Motile Neural Crest Cells and Affect Craniofacial Development in Xenopus laevis.

Wolf-Hirschhorn Syndrome (WHS) is a human developmental disorder arising from a hemizygous perturbation, typically a microdeletion, on the short arm of chromosome four. In addition to pronounced intellectual disability, seizures, and delayed growth, WHS presents with a characteristic facial dysmorphism and varying prevalence of microcephaly, micrognathia, cartilage malformation in the ear and nose, and facial asymmetries. These affected craniofacial tissues all derive from a shared embryonic precursor, the cranial neural crest (CNC), inviting the hypothesis that one or more WHS-affected genes may be critical regulators of neural crest development or migration. To explore this, we characterized expression of multiple genes within or immediately proximal to defined WHS critical regions, across the span of craniofacial development in the vertebrate model system Xenopus laevis. This subset of genes, whsc1, whsc2, letm1, and tacc3, are diverse in their currently-elucidated cellular functions; yet we find that their expression demonstrates shared tissue-specific enrichment within the anterior neural tube, migratory neural crest, and later craniofacial structures. We examine the ramifications of this by characterizing craniofacial development and neural crest migration following individual gene depletion. We observe that several WHS-associated genes significantly impact facial patterning, cartilage formation, neural crest motility in vivo and in vitro, and can separately contribute to forebrain scaling. Thus, we have determined that numerous genes within and surrounding the defined WHS critical regions potently impact craniofacial patterning, suggesting their role in WHS presentation may stem from essential functions during neural crest-derived tissue formation.

Characterization of Xenopus laevis guanine deaminase reveals new insights for its expression and function in the embryonic kidney.

BACKGROUND: The mammalian guanine deaminase (GDA), called cypin, is important for proper neural development, by regulating dendritic arborization through modulation of microtubule (MT) dynamics. Additionally, cypin can promote MT assembly in vitro. However, it has never been tested whether cypin (or other GDA orthologs) binds to MTs or modulates MT dynamics. Here, we address these questions and characterize Xenopus laevis GDA (Gda) for the first time during embryonic development. RESULTS: We find that exogenously expressed human cypin and Gda both display a cytosolic distribution in primary embryonic cells. Furthermore, while expression of human cypin can promote MT polymerization, Xenopus Gda has no effect. Additionally, we find that the tubulin-binding collapsin response mediator protein (CRMP) homology domain is only partially conserved between cypin and Gda. This likely explains the divergence in function, as we discovered that the cypin region containing the CRMP homology and PDZ-binding domain is necessary for regulating MT dynamics. Finally, we observed that gda is strongly expressed in the kidneys during late embryonic development, although it does not appear to be critical for kidney development. CONCLUSIONS: Together, these results suggest that GDA has diverged in function between mammals and amphibians, and that mammalian GDA plays an indirect role in regulating MT dynamics. Developmental Dynamics 248:296-305, 2019. © 2019 Wiley Periodicals, Inc.

The origin and migration of the dissolved sulfate from precipitation in Seoul, Korea.

This study incorporated stable isotope analyses with chemical analyses to determine the origin and migration of sulfur sources in East Asia, and these findings were compared with our decadal research from 2000 to 2001 and 2002 to 2003. The multiple sulfur isotope composition (32S, 33S and 34S) of the dissolved sulfate in precipitation was first measured from 2011 to 2013 in Seoul, South Korea. The δ34Snss values were -1.1‰ to 7.9‰ (avg. 3.6‰), strongly suggesting that sulfur derived from the combustion of Chinese coal is the predominant source of sulfate in the Seoul region. Low NO3/SO42- ratios in the precipitation samples indicated an insignificant effect of sulfur from vehicle exhaust. The seasonal variation of δ34Snss values appears to be caused by increasing biogenic sulfur activity during the spring and summer seasons. The some Δ33S values (0.13‰-0.16‰) measured in the three samples were sufficiently small; thus, whether these values can be attributed to mass-independent fractionation remains unclear. Measuring the Δ33S anomalies in dissolved sulfate provides valuable insights for identifying the sources of sulfur transferred from the stratosphere to the troposphere and upper troposphere.

Assembly and turnover of neurofilaments in growing axonal neurites.

Neurofilaments (NFs) are thought to provide stability to the axon. We examined NF dynamics within axonal neurites of NB2a/d1 neuroblastoma by transient transfection with green fluorescent protein-tagged NF-heavy (GFP-H) under the control of a tetracycline-inducible promoter. Immunofluorescent and biochemical analyses demonstrated that GFP-H expressed early during neurite outgrowth associated with a population of centrally-situated, highly-phosphorylated crosslinked NFs along the length of axonal neurites ('bundled NFs'). By contrast, GFP-H expressed after considerable neurite outgrowth displayed markedly reduced association with bundled NFs and was instead more evenly distributed throughout the axon. This differential localization was maintained for up to 2 weeks in culture. Once considerable neurite outgrowth had progressed, GFP that had previously associated with the NF bundle during early expression was irreversibly depleted by photobleaching. Cessation of expression allowed monitoring of NF turnover. GFP-H associated bundled NFs underwent slower decay than GFP-H associated with surrounding, less-phosphorylated NFs. Notably, GFP associated with bundled NFs underwent similar decay rates within the core and edges of this bundle. These results are consistent with previous demonstration of a resident NF population within axonal neurites, but suggest that this population is more dynamic than previously considered.

Robot-Embodied Neuronal Networks as an Interactive Model of Learning.

BACKGROUND AND OBJECTIVE: The reductionist approach of neuronal cell culture has been useful for analyses of synaptic signaling. Murine cortical neurons in culture spontaneously form an ex vivo network capable of transmitting complex signals, and have been useful for analyses of several fundamental aspects of neuronal development hitherto difficult to clarify in situ. However, these networks lack the ability to receive and respond to sensory input from the environment as do neurons in vivo. Establishment of these networks in culture chambers containing multi-electrode arrays allows recording of synaptic activity as well as stimulation. METHOD: This article describes the embodiment of ex vivo neuronal networks neurons in a closed-loop cybernetic system, consisting of digitized video signals as sensory input and a robot arm as motor output. RESULTS: In this system, the neuronal network essentially functions as a simple central nervous system. This embodied network displays the ability to track a target in a naturalistic environment. These findings underscore that ex vivo neuronal networks can respond to sensory input and direct motor output. CONCLUSION: These analyses may contribute to optimization of neuronal-computer interfaces for perceptive and locomotive prosthetic applications. Ex vivo networks display critical alterations in signal patterns following treatment with subcytotoxic concentrations of amyloid-beta. Future studies including comparison of tracking accuracy of embodied networks prepared from mice harboring key mutations with those from normal mice, accompanied with exposure to Abeta and/or other neurotoxins, may provide a useful model system for monitoring subtle impairment of neuronal function as well as normal and abnormal development.

Omega-3 Hastens and Omega-6 Delays the Progression of Neuropathology in a Murine Model of Familial ALS.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a progressive disease of motor neurons that has no cure or effective treatment. Any approach that could sustain minor motor function during terminal stages would improve quality of life. OBJECTIVE: We examined the impact of omega-3 (Ω-3) and Ω-6, on motor neuron function in mice expressing mutant human superoxide dismutase-1 (SOD-1), which dominantly confers familial ALS and induces a similar sequence of motor neuron decline and eventual death when expressed in mice. METHOD: Mice received standard diets supplemented with equivalent amounts of Ω-3 and Ω-6 or a 10x increase in Ω-6 with no change in Ω-3 commencing at 4 weeks of age. Motor function and biochemical/histological parameters were assayed by standard methodologies. RESULTS: Supplementation with equivalent Ω-3 and Ω-6 hastened motor neuron pathology and death, while 10x Ω-6 with no change in Ω-3 significantly delayed motor neuron pathology, including preservation of minor motor neuron function during the terminal stage. CONCLUSION: In the absence of a cure or treatment, affected individuals may resort to popular nutritional supplements such as Ω-3 as a form of "self-medication". However, our findings and those of other laboratories indicate that such an approach could be harmful. Our findings suggest that a critical balance of Ω-6 and Ω-3 may temporarily preserve motor neuron function during the terminal stages of ALS, which could provide a substantial improvement in quality of life for affected individuals and their caregivers.

Insufficient developmental excitatory neuronal activity fails to foster establishment of normal levels of inhibitory neuronal activity.

The nervous system is composed of excitatory and inhibitory neurons. One major class of inhibitory neurons release the neurotransmitter γ-Aminobutyric acid (GABA). GABAergic inhibitory activity maintains the balance that is disrupted in conditions such as epilepsy. At least some GABAergic neurons are initially excitatory and undergo a developmental conversion to convert to inhibitory neurons. The mechanism(s) behind this conversion are thought to include a critical developmental increase in excitatory activity. To test this hypothesis, we subjected ex vivo developing neuronal networks on multi-electrode arrays to various stimulation and pharmacological regimens. Synaptic activity of networks initially consists of epileptiform-like high-amplitude individual "spikes", which convert to organized bursts of activity over the course of approximately 1 month. Stimulation of networks with a digitized synaptic signal for 5days hastened the decrease of epileptiform activity. By contrast, stimulation for a single day delayed the appearance of bursts and instead increased epileptiform signaling. GABA treatment reduced total signals in unstimulated networks and networks stimulated for 5days, but instead increased signaling in networks stimulated for 1day. This increase was prevented by co-treatment with (2R)-amino-5-phosphonopentanoate and 6-cyano-7-nitroquinoxaline-2,3-dione, confirming that GABA invoked excitatory activity in networks stimulated for 1day. Glutamate increased signals in networks subjected to all stimulation regimens; the GABA receptor antagonist bicuculline prevented this increase only in networks stimulated for 1day. These latter findings are consistent with the induction of so-called "mixed" synapses (which release a combination of excitatory and inhibitory neurotransmitters) in networks stimulated for 1day, and support the hypothesis that a critical level of excitatory activity fosters the developmental transition of GABAergic neurons from excitatory to inhibitory.

Social interaction attenuates the extent of secondary neuronal damage following closed head injury in mice.

Recovery following Traumatic Brain Injury (TBI) can vary tremendously among individuals. Lifestyle following injury, including differential social interactions, may modulate the extent of secondary injury following TBI. To examine this possibility under controlled conditions, closed head injury (CHI) was induced in C57Bl6 mice using a standardized weight drop device after which mice were either housed in isolation or with their original cagemates ("socially-housed") for 4 weeks. CHI transiently impaired novel object recognition (NOR) in both isolated and social mice, confirming physical and functional injury. By contrast, Y maze navigation was impaired in isolated but not social mice at 1-4 weeks post CHI. CHI increased excitotoxic signaling in hippocampal slices from all mice, which was transiently exacerbated by isolation at 2 weeks post CHI. CHI slightly increased reactive oxygen species and did not alter levels of amyloid beta (Abeta), total or phospho-tau, total or phosphorylated neurofilaments. CHI increased serum corticosterone in both groups, which was exacerbated by isolation. These findings support the hypothesis that socialization may attenuate secondary damage following TBI. In addition, a dominance hierarchy was noted among socially-housed mice, in which the most submissive mouse displayed indices of stress in the above analyses that were statistically identical to those observed for isolated mice. This latter finding underscores that the nature and extent of social interaction may need to vary among individuals to provide therapeutic benefit.

Additive Impairment of Synaptic Signaling in Cultured Cortical Neurons by Exogenously-Applied Oligomerized Amyloid-ß and Airborne Nanoparticles Generated during Photocopying.

Photocopying in offices and printing centers releases nanoparticles that can reach the brain following inhalation. We examined whether subcytotoxic levels of airborne photocopy-emitted nanoparticles could potentiate perturbation of synaptic signaling in cultured neurons following exposure to amyloid-ß (Aß). Signaling was only transiently inhibited by Aß or nanoparticles individually, but remained statistically reduced in cultures receiving both after 24 h. In vitro and in vivo studies with copier emitted nanoparticles have consistently demonstrated inflammation, oxidative stress, and cytotoxicity. Since Aß can accumulate years before cognitive decline, subcytotoxic levels of nanoparticles are one factor that could potentiate Aß-induced impairment of synaptic activity during these early stages.

Detecting changes in arthritic fibroblast-like synoviocytes using atomic force microscopy.

The morphological and quantitative differences between arthritic fibroblast-like synoviocytes (FLS) and normal FLS were determined as an effective diagnostic tool for rheumatoid arthritis (RA), and confirmed using atomic force microscopy (AFM). Collagen-induced arthritic (CIA) mice and normal mice were prepared and FLS were isolated by enzymatic digestion from the synovial tissue of sacrificed mice at 5-week and 8-week pathogenesis periods. Analysis of cell morphology using AFM revealed that the surface roughness around the nucleus and around the branched cytoplasm was significantly higher in CIA FLS (P < 0.05) than that in normal FLS. In addition, the roughness of two different sites on the arthritic FLS increased with an increase in the duration of pathogenesis. These results strongly suggest that AFM can be widely used as a diagnostic tool in cytopathology to detect the early signs of RA and various others diseases at the intercellular level.

Early expression of the high molecular weight neurofilament subunit attenuates axonal neurite outgrowth.

Phospho-dependent interactions of the C-terminal region of the high molecular weight NF subunit (NF-H) with each other and with other cytoskeletal elements stabilize the axonal cytoskeleton and contribute to an increase in axonal caliber. The same kinase cascades that mediate axonal pathfinding via growth cone dynamics are those that foster NF-mediated axonal stabilization, yet there is a developmental delay in the accumulation of NF C-terminal phosphorylation. Moreover, the phospho-mediated C-terminal NF-H interactions that stabilize the axonal cytoskeleton also inhibit axonal elongation. We hypothesized that a delay in expression and/or accumulation of NF-H within developing axons is essential to allow axonal elongation and pathfinding. We tested this hypothesis in differentiating NB2a/d1 cells. The first 3 days of differentiation of NB2a/d1 cells is normally accompanied by rapid elongation of axonal neurites. This period is followed by the accumulation of C-terminally phosphorylated NF-H, cessation of axonal elongation and an increase in axonal caliber. Herein, overexpression of GFP-tagged NF-H simultaneously with induction of differentiation fostered accumulation of C-terminally phosphorylated NF-H within developing axonal neurites within 48hr, which was accompanied by retardation of axonal elongation and a hastened increase in caliber. These effects were prevented by treatment with inhibitors of kinases that mediate the association of NFs with other cytoskeletal elements. Overexpression of GFP-NF-H lacking the C-terminal 187 amino acids (which mediate NF-NF interactions) did not retard elongation nor increase caliber. These findings support the hypothesis that a developmental delay in NF-H C-terminal phosphorylation is essential to allow appropriate axonal elongation prior to stabilization.

The high molecular weight neurofilament subunit plays an essential role in axonal outgrowth and stabilization.

Neurofilaments (NFs) are thought to provide structural support to mature axons via crosslinking of cytoskeletal elements mediated by the C-terminal region of the high molecular weight NF subunit (NF-H). Herein, we inhibited NF-H expression in differentiating mouse NB2a/d1 cells with shRNA directed against murine NF-H without affecting other NF subunits, microtubules or actin. shRNA-mediated NF-H knockdown not only in compromised of late-stage axonal neurite stabilization but also compromised early stages of axonal neurite elongation. Expression of exogenous rat NF-H was able to compensate for knockdown of endogenous NF-H and restored the development and stabilization of axonal neurites. This rescue was prevented by simultaneous treatment with shRNA that inhibited both rat and murine NF-H, or by expression of exogenous rat NF-H lacking the C-terminal sidearm during knockdown of endogenous NF-H. Demonstration of a role for NF-H in the early stages of axonal elaboration suggests that axonal stabilization is not delayed until synaptogenesis, but rather that the developing axon undergoes sequential NF-H-mediated stabilization along its length in a proximal-distal manner, which supports continued pathfinding in distal, unstabilized regions.

Synaptic signal streams generated by ex vivo neuronal networks contain non-random, complex patterns.

Cultured embryonic neurons develop functional networks that transmit synaptic signals over multiple sequentially connected neurons as revealed by multi-electrode arrays (MEAs) embedded within the culture dish. Signal streams of ex vivo networks contain spikes and bursts of varying amplitude and duration. Despite the random interactions inherent in dissociated cultures, neurons are capable of establishing functional ex vivo networks that transmit signals among synaptically connected neurons, undergo developmental maturation, and respond to exogenous stimulation by alterations in signal patterns. These characteristics indicate that a considerable degree of organization is an inherent property of neurons. We demonstrate herein that (1) certain signal types occur more frequently than others, (2) the predominant signal types change during and following maturation, (3) signal predominance is dependent upon inhibitory activity, and (4) certain signals preferentially follow others in a non-reciprocal manner. These findings indicate that the elaboration of complex signal streams comprised of a non-random distribution of signal patterns is an emergent property of ex vivo neuronal networks.

Divergent and convergent roles for kinases and phosphatases in neurofilament dynamics.

C-terminal neurofilament phosphorylation mediates cation-dependent self-association leading to neurofilament incorporation into the stationary axonal cytoskeleton. Multiple kinases phosphorylate the C-terminal domains of the heavy neurofilament subunit (NF-H), including cyclin-dependent protein kinase 5 (CDK5), mitogen-activated protein kinases (MAPKs), casein kinase 1 and 2 (CK1 and CK2) and glycogen synthase kinase 3ß (GSK3ß). The respective contributions of these kinases have been confounded because they phosphorylate multiple substrates in addition to neurofilaments and display extensive interaction. Herein, differentiated NB2a/d1 cells were transfected with constructs expressing GFP-tagged NF-H, isolated NF-H sidearms and NF-H lacking the distal-most 187 amino acids. Cultures were treated with roscovitine, PD98059, Li(+), D4476, tetrabromobenzotriazole and calyculin, which are active against CDK5, MKK1 (also known as MAP2K1), GSK3ß, CK1, CK2 and protein phosphatase 1 (PP1), respectively. Sequential phosphorylation by CDK5 and GSK3ß mediated the neurofilament-neurofilament associations. The MAPK pathway (i.e. MKK1 to ERK1/2) was found to downregulate GSK3ß, and CK1 activated PP1, both of which promoted axonal transport and restricted neurofilament-neurofilament associations to axonal neurites. The MAPK pathway and CDK5, but not CK1 and GSK3ß, inhibited neurofilament proteolysis. These findings indicate that phosphorylation of neurofilaments by the proline-directed MAPK pathway and CDK5 counterbalance the impact of phosphorylation of neurofilaments by the non-proline-directed CK1 and GSK3ß.