Hydrogel-Encapsulated Biofilm Inhibitors Abrogate the Cariogenic Activity of Streptococcus mutans.

We designed and synthesized analogues of a previously identified biofilm inhibitor IIIC5 to improve solubility, retain inhibitory activities, and to facilitate encapsulation into pH-responsive hydrogel microparticles. The optimized lead compound HA5 showed improved solubility of 120.09 µg/mL, inhibited Streptococcus mutans biofilm with an IC50 value of 6.42 µM, and did not affect the growth of oral commensal species up to a 15-fold higher concentration. The cocrystal structure of HA5 with GtfB catalytic domain determined at 2.35 Å resolution revealed its active site interactions. The ability of HA5 to inhibit S. mutans Gtfs and to reduce glucan production has been demonstrated. The hydrogel-encapsulated biofilm inhibitor (HEBI), generated by encapsulating HA5 in hydrogel, selectively inhibited S. mutans biofilms like HA5. Treatment of S. mutans-infected rats with HA5 or HEBI resulted in a significant reduction in buccal, sulcal, and proximal dental caries compared to untreated, infected rats.

Generation of genomic-integration-free human induced pluripotent stem cells and the derived cardiomyocytes of X-linked dilated cardiomyopathy from DMD gene mutation.

We derived an integration-free induced pluripotent stem cell (iPSC) line from the peripheral blood mononuclear cells (PBMCs) of a 23-year-old male patient. This patient carries a 5' splice site point mutation in intron 1 (c.31+1G>A) of the dystrophin gene, a mutation associated with X-linked dilated cardiomyopathy (XLDCM). Sendai virus was used to reprogram the PBMCs and deliver OCT3/4, SOX2, c-MYC, and KLF4 factors. The iPSC line (HKUi002-A) generated preserved the mutation, expressed common pluripotency markers, differentiated into three germ layers in vivo, and exhibited a normal karyotype. Further differentiation into cardiomyocytes enables the study of the disease mechanisms of XLDCM.

The disruptive effects of pain on multitasking in a virtual errands task.

BACKGROUND AND AIMS: Pain is known to have a disruptive effect on cognitive performance, but prior studies have used highly constrained laboratory tasks that lack ecological validity. In everyday life people are required to complete more complex sets of tasks, prioritising task completion and recalling lists of tasks which need to be completed, and these tasks continue to be attempted during episodes or states of pain. The present study therefore examined the impact of thermal induced pain on a simulated errand task. METHODS: Fifty-five healthy adults (36 female) performed the Edinburgh Virtual Errands Task (EVET) either during a painful thermal sensation or with no concurrent pain. Participants also completed the Experience of Cognitive Intrusion of Pain (ECIP) questionnaire to measure their self-reported cognitive impact of pain in general life. RESULTS: Participants who completed the EVET task in pain and who self-reported high intrusion of pain made significantly more errors than those who reported lower intrusion on the ECIP. CONCLUSIONS: Findings here support the growing literature that suggests that pain has a significant impact on cognitive performance. Furthermore, these findings support the developing literature suggesting that this relationship is complex when considering real world cognition, and that self-report on the ECIP relates well to performance on a task designed to reflect the complexities of everyday living. IMPLICATIONS: If extrapolated to chronic pain populations, these data suggest that pain during complex multitasking performance may have a significant impact on the number of errors made. For people highly vulnerable to cognitive intrusion by pain, this may result in errors such as selecting the wrong location or item to perform tasks, or forgetting to perform these tasks at the correct time. If these findings are shown to extend to chronic pain populations then occupational support to manage complex task performance, using for example diaries/electronic reminders, may help to improve everyday abilities.

Antiviral effect of forsythoside A from Forsythia suspensa (Thunb.) Vahl fruit against influenza A virus through reduction of viral M1 protein.

ETHNOPHARMACOLOGICAL RELEVANCE: Yinqiaosan is a classical traditional Chinese medicine formula, which has been used to treat respiratory diseases since ancient China. It consists of nine herbs and among them, Forsythia suspensa (Thunb.) Vahl fruit is one of the major herbal components. Despite the long history of Yinqiaosan, the active compounds and the mechanisms of action of this formula remain elusive. AIM OF THE STUDY: The present study aimed to examine the suppressive effect of Yinqiaosan on influenza virus and to identify the active components in the formula targeting influenza. MATERIALS AND METHODS: Anti-influenza virus effect of Yinqiaosan was assessed by tissue culture infective dose assay, and was also tested in an in vivo mouse model. Active compound from the formula was identified with a bioactivity-guided fractionation scheme. The potential mode of action of the compound was further investigated by identifying the host cell signaling pathways and viral protein production using in vitro cell culture models. RESULTS: Our results showed that forsythoside A from Forsythia suspensa (Thunb.) Vahl fruit, a major herbal component in Yinqiaosan, reduced the viral titers of different influenza virus subtypes in cell cultures and increased the survival rate of the mice in an in vivo influenza virus infection model. Further experiments on the mode of action of forsythoside A showed that it reduced the influenza M1 protein, which in turn intervened the budding process of the newly formed virions and eventually limited the virus spread. CONCLUSION: Results of our present study provides scientific evidence to support to the application of a traditional herbal formula. We also identify novel candidate compound for future drug development against influenza virus.

Looking back at the stare-in-the-crowd effect: staring eyes do not capture attention in visual search.

The stare-in-the crowd effect refers to the finding that a visual search for a target of staring eyes among averted-eyes distracters is more efficient than the search for an averted-eyes target among staring distracters. This finding could indicate that staring eyes are prioritized in the processing of the search array so that attention is more likely to be directed to their location than to any other. However, visual search is a complex process, which not only depends upon the properties of the target, but also the similarity between the target of the search and the distractor items and between the distractor items themselves. Across five experiments, we show that the search asymmetry diagnostic of the stare-in-the-crowd effect is more likely to be the result of a failure to control for the similarity among distracting items between the two critical search conditions rather than any special attention-grabbing property of staring gazes. Our results suggest that, contrary to results reported in the literature, staring gazes are not prioritized by attention in visual search.

A role for protein phosphatase 2A in regulating p38 mitogen activated protein kinase activation and tumor necrosis factor-alpha expression during influenza virus infection.

Influenza viruses of avian origin continue to pose pandemic threats to human health. Some of the H5N1 and H9N2 virus subtypes induce markedly elevated cytokine levels when compared with the seasonal H1N1 virus. We previously showed that H5N1/97 hyperinduces tumor necrosis factor (TNF)-alpha through p38 mitogen activated protein kinase (MAPK). However, the detailed mechanisms of p38MAPK activation and TNF-alpha hyperinduction following influenza virus infections are not known. Negative feedback regulations of cytokine expression play important roles in avoiding overwhelming production of proinflammatory cytokines. Here we hypothesize that protein phosphatases are involved in the regulation of cytokine expressions during influenza virus infection. We investigated the roles of protein phosphatases including MAPK phosphatase-1 (MKP-1) and protein phosphatase type 2A (PP2A) in modulating p38MAPK activation and downstream TNF-alpha expressions in primary human monocyte-derived macrophages (PBMac) infected with H9N2/G1 or H1N1 influenza virus. We demonstrate that H9N2/G1 virus activated p38MAPK and hyperinduced TNF-alpha production in PBMac when compared with H1N1 virus. H9N2/G1 induced PP2A activity in PBMac and, with the treatment of a PP2A inhibitor, p38MAPK phosphorylation and TNF-alpha production were further increased in the virus-infected macrophages. However, H9N2/G1 did not induce the expression of PP2A indicating that the activation of PP2A is not mediated by p38MAPK in virus-infected PBMac. On the other hand, PP2A may not be the targets of H9N2/G1 in the upstream of p38MAPK signaling pathways since H1N1 also induced PP2A activation in primary macrophages. Our results may provide new insights into the control of cytokine dysregulation.

The impact of working memory load on task execution and online plan adjustment during multitasking in a virtual environment.

Three experiments investigated the impact of working memory load on online plan adjustment during a test of multitasking in young, nonexpert, adult participants. Multitasking was assessed using the Edinburgh Virtual Errands Test (EVET). Participants were asked to memorize either good or poor plans for performing multiple errands and were assessed both on task completion and on the extent to which they modified their plans during EVET performance. EVET was performed twice, with and without a secondary task loading a component of working memory. In Experiment 1, articulatory suppression was used to load the phonological loop. In Experiment 2, oral random generation was used to load executive functions. In Experiment 3, spatial working memory was loaded with an auditory spatial localization task. EVET performance for both good- and poor-planning groups was disrupted by random generation and sound localization, but not by articulatory suppression. Additionally, people given a poor plan were able to overcome this initial disadvantage by modifying their plans online. It was concluded that, in addition to executive functions, multiple errands performance draws heavily on spatial, but not verbal, working memory resources but can be successfully completed on the basis of modifying plans online, despite a secondary task load.

Event-based prospective remembering in a virtual world.

Most laboratory-based prospective memory (PM) paradigms pose problems that are very different from those encountered in the real world. Several PM studies have reported conflicting results when comparing laboratory- with naturalistic-based studies (e.g., Bailey, Henry, Rendell, Phillips, & Kliegel, 2010 ). One key contrast is that for the former, how and when the PM cue is encountered typically is determined by the experimenter, whereas in the latter case, cue availability is determined by participant actions. However, participant-driven access to the cue has not been examined in laboratory studies focused on healthy young adults, and its relationship with planned intentions is poorly understood. Here we report a study of PM performance in a controlled, laboratory setting, but with participant-driven actions leading to the availability of the PM cue. This uses a novel PM methodology based upon analysis of participant movements as they attempted a series of errands in a large virtual building on the computer screen. A PM failure was identified as a situation in which a participant entered and exited the "cue" area outside an errand related room without performing the required errand whilst still successfully remembering that errand post test. Additional individual difference measures assessed retrospective and working memory capacity, planning ability and PM. Multiple regression analysis showed that the independent measures of verbal working memory span, planning ability, and PM were significant predictors of PM failure. Correlational analyses with measures of planning suggest that sticking with an original plan (good or bad) is related to better overall PM performance.

Traffic Impacts on PM(2.5) Air Quality in Nairobi, Kenya.

Motor vehicle traffic is an important source of particulate pollution in cities of the developing world, where rapid growth, coupled with a lack of effective transport and land use planning, may result in harmful levels of fine particles (PM(2.5)) in the air. However, a lack of air monitoring data hinders health impact assessments and the development of transportation and land use policies that could reduce health burdens due to outdoor air pollution. To address this important need, a study of traffic-related PM(2.5) was carried out in the city of Nairobi, Kenya, a model city for sub-Saharan Africa, in July 2009. Sampling was carried out using portable filter-based air samplers carried in backpacks by technicians on weekdays over two weeks at several sites in and around Nairobi ranging from high-traffic roadways to rural background. Mean daytime concentrations of PM(2.5) ranged from 10.7 at the rural background site to 98.1 µg/m(3) on a sidewalk in the central business district. Horizontal dispersion measurements demonstrated a decrease in PM(2.5) concentration from 128.7 to 18.7 µg/m(3) over 100 meters downwind of a major intersection in Nairobi. A vertical dispersion experiment revealed a decrease from 119.5 µg/m(3) at street level to 42.8 µg/m(3) on a third-floor rooftop in the central business district. Though not directly comparable to air quality guidelines, which are based on 24-hour or annual averages, the urban concentrations we observed raise concern with regard to public health and related policy. Taken together with survey data on commuting patterns within Nairobi, these results suggest that many Nairobi residents are exposed on a regular basis to elevated concentrations of fine particle air pollution, with potentially serious long-term implications for health.

Multitasking: multiple, domain-specific cognitive functions in a virtual environment.

Multitasking among three or more different tasks is a ubiquitous requirement of everyday cognition, yet rarely is it addressed in research on healthy adults who have had no specific training in multitasking skills. Participants completed a set of diverse subtasks within a simulated shopping mall and office environment, the Edinburgh Virtual Errands Test (EVET). The aim was to investigate how different cognitive functions, such as planning, retrospective and prospective memory, and visuospatial and verbal working memory, contribute to everyday multitasking. Subtasks were chosen to be diverse, and predictions were derived from a statistical model of everyday multitasking impairments associated with frontal-lobe lesions (Burgess, Veitch, de Lacy Costello, & Shallice, 2000b). Multiple regression indicated significant independent contributions from measures of retrospective memory, visuospatial working memory, and online planning, but not from independent measures of prospective memory or verbal working memory. Structural equation modelling showed that the best fit to the data arose from three underlying constructs, with Memory and Planning having a weak link, but with both having a strong directional pathway to an Intent construct that reflected implementation of intentions. Participants who followed their preprepared plan achieved higher scores than those who altered their plan during multitask performance. This was true regardless of whether the plan was efficient or poor. These results substantially develop and extend the Burgess et al. (2000b) model to healthy adults and yield new insight into the poorly understood area of everyday multitasking. The findings also point to the utility of using virtual environments for investigating this form of complex human cognition.

Preliminary evidence that glucose ingestion facilitates prospective memory performance.

Previous research has found that the ingestion of glucose boosts task performance in the memory domain (including tasks tapping episodic, semantic, and working memory). The present pilot study tested the hypothesis that glucose ingestion would enhance performance on a test of prospective memory. In a between-subjects design, 56 adults ranging from 17 to 80 years of age performed a computerized prospective memory task and an attention (filler) task after 25 g of glucose or a sweetness-matched placebo. Blood glucose measurements were also taken to assess the impact of individual differences on glucose regulation. After the drink containing glucose, cognitive facilitation was observed on the prospective memory task after excluding subjects with impaired fasting glucose level. Specifically, subjects receiving glucose were 19% more accurate than subjects receiving a placebo, a trend that was marginally nonsignificant, F1,41 = 3.4, P = .07, but that had a medium effect size, d = 0.58. Subjects receiving glucose were also significantly faster on the prospective memory task, F1,35 = 4.8, P < .05, d = 0.6. In addition, elevated baseline blood glucose (indicative of poor glucose regulation) was associated with slower prospective memory responding, F1,35 = 4.4, P < .05, d = 0.57. These data add to the growing body of evidence suggesting that both memory and executive functioning can benefit from the increased provision of glucose to the brain.

Isolation and identification of anti-inflammatory constituents from Ligusticum chuanxiong and their underlying mechanisms of action on microglia.

Stroke is the third most common cause of death worldwide. Recent findings showed that the severity of cerebrovascular diseases including ischemic stroke correlates with inflammation mediated responses in the neural cells. During ischemia, inflammatory mediators including tumor necrosis factor-alpha (TNF-α) and nitric oxide are produced by microglia, which play a central role in the pathogenesis of the disease. Ligusticum chuanxiong (LCX) is a commonly used traditional Chinese medicine (TCM) for empiric treatment of cerebrovascular and cardiovascular diseases for many centuries. By applying a bioactivity-guided fractionation scheme, two compounds with inhibition on neuroinflammation were isolated from LCX. Using chromatographic and spectrometric methods, they were identified to be senkyunolide A and Z-ligustilide. They could inhibit the production of proinflammatory mediators in lipopolysaccharide (LPS)-stimulated murine BV-2 microglial cells and human peripheral blood monocyte derived macrophages. In addition, both compounds protected Neuro-2a cells from neuroinflammatory toxicity induced by the conditioned culture media produced by LPS-stimulated BV-2 cells. The underlying mechanisms of action of senkyunolide A were further delineated. Its inhibitory effects were shown to be independent of the phosphorylation of mitogen-activated protein kinases (MAPK) and translocation of nuclear factor kappa B (NF-κB). However, senkyunolide A could increase the degradation of TNF-α mRNA and reduce its half life by 43%. In conclusion, bioactivity-guided fractionation is an effective way of isolating bioactive compounds from medicinal herbs. In addition, senkyunolide A and Z-ligustilide isolated from LCX may be considered as potential complementary drug candidates for treating inflammatory processes associated with cerebrovascular diseases.

Investigating multitasking in high-functioning adolescents with autism spectrum disorders using the Virtual Errands Task.

Using a modified version of the Virtual Errands Task (VET; McGeorge et al. in Presence-Teleop Virtual Environ 10(4):375-383, 2001), we investigated the executive ability of multitasking in 18 high-functioning adolescents with ASD and 18 typically developing adolescents. The VET requires multitasking (Law et al. in Acta Psychol 122(1):27-44, 2006) because there is a limited amount of time in which to complete the errands. ANCOVA revealed that the ASD group completed fewer tasks, broke more rules and rigidly followed the task list in the order of presentation. Our findings suggest that executive problems of planning inflexibility, inhibition, as well as difficulties with prospective memory (remembering to carry out intentions) may lie behind multitasking difficulties in ASD.

Cellular response to influenza virus infection: a potential role for autophagy in CXCL10 and interferon-alpha induction.

Historically, influenza pandemics have arisen from avian influenza viruses. Avian influenza viruses H5N1 and H9N2 are potential pandemic candidates. Infection of humans with the highly pathogenic avian influenza H5N1 virus is associated with a mortality in excess of 60%, which has been attributed to dysregulation of the cytokine system. Human macrophages and epithelial cells infected with some genotypes of H5N1 and H9N2 viruses express markedly elevated cytokine and chemokine levels when compared with seasonal influenza A subtype H1N1 virus. The mechanisms underlying this cytokine and chemokine hyperinduction are not fully elucidated. In the present study, we demonstrate that autophagy, a tightly regulated homeostatic process for self-digestion of unwanted cellular subcomponents, plays a role in cytokine induction. Autophagy is induced to a greater extent by H9N2/G1, in association with cytokine hyperinduction, compared with H1N1 and the novel pandemic swine-origin influenza A/H1N1 viruses. Using 3-methyladenine to inhibit autophagy and small interfering RNA to silence the autophagy gene, Atg5, we further show that autophagic responses play a role in influenza virus-induced CXCL10 and interferon-alpha expression in primary human blood macrophages. Our results provide new insights into the pathogenic mechanisms of avian influenza viruses.

Differential replication of avian influenza H9N2 viruses in human alveolar epithelial A549 cells.

Avian influenza virus H9N2 isolates cause a mild influenza-like illness in humans. However, the pathogenesis of the H9N2 subtypes in human remains to be investigated. Using a human alveolar epithelial cell line A549 as host, we found that A/Quail/Hong Kong/G1/97 (H9N2/G1), which shares 6 viral "internal genes" with the lethal A/Hong Kong/156/97 (H5N1/97) virus, replicates efficiently whereas other H9N2 viruses, A/Duck/Hong Kong/Y280/97 (H9N2/Y280) and A/Chicken/Hong Kong/G9/97 (H9N2/G9), replicate poorly. Interestingly, we found that there is a difference in the translation of viral protein but not in the infectivity or transcription of viral genes of these H9N2 viruses in the infected cells. This difference may possibly be explained by H9N2/G1 being more efficient on viral protein production in specific cell types. These findings suggest that the H9N2/G1 virus like its counterpart H5N1/97 may be better adapted to the human host and replicates efficiently in human alveolar epithelial cells.

A role for protein kinase PKR in the mediation of Epstein-Barr virus latent membrane protein-1-induced IL-6 and IL-10 expression.

Expression of Epstein-Barr virus-encoded oncogenic latent membrane protein 1 (LMP1) has been substantially associated with tumorigenic transformation in the virus-infected cells. The pathogenic complexity of LMP1 is partly due to the cytokine dysregulation including IL-6 and IL-10 in perturbing the host immune responses. Here we have identified an important signaling event mediated by a dsRNA-dependent serine/threonine protein kinase, PKR, in regulating LMP1-induced IL-6 and IL-10 expression. We first demonstrated that PKR plays a significant role in mediating LMP1-induced cytokine expression by using a PKR inhibitor 2-aminopurine, and the specific role of PKR involved was confirmed by the use of siRNA oligos targeting PKR and/or a dominant-negative PKR mutant. We next revealed that PKR activity mediates LMP1-enhanced NF-kappaB nuclear translocation resulting in cytokine induction. We further demonstrated at the chromatin level that LMP1 can significantly elevate the phosphorylation of histone H3 on serine 10 (Ser 10), and the process was dependent on PKR activity. Our findings thus suggest that PKR plays an important role in mediating the cytokine gene expression induced by LMP1 through NF-kappaB activation and histone H3 Ser 10 phosphorylation.

A novel small-molecule inhibitor of the avian influenza H5N1 virus determined through computational screening against the neuraminidase.

Computational molecular docking provides an efficient and innovative approach to examine small molecule and protein interactions. We have utilized this method to identify potential inhibitors of the H5N1 neuraminidase protein. Of the 20 compounds tested, 4-(4-((3-(2-amino-4-hydroxy-6-methyl-5-pyrimidinyl)propyl)amino)phenyl)-1-chloro-3-buten-2-one (1) (NSC89853) demonstrated the ability to inhibit viral replication at a level comparable to the known neuraminidase inhibitor oseltamivir. Compound 1 demonstrated efficacy across a number of cell-lines assays and in both the H1N1 and H5N1 viruses. The predicted binding of 1 to the known H5N1 neuraminidase structure indicates a binding interface largely nonoverlapping with that of oseltamivir or another neuraminidase inhibitor zanamivir. These results indicate that 1 or similar molecules would remain effective in the presence of virus mutations conferring resistance to either oseltamivir or zanamivir and also vice versa.

Attention capture by faces.

We report three experiments that investigate whether faces are capable of capturing attention when in competition with other non-face objects. In Experiment 1a participants took longer to decide that an array of objects contained a butterfly target when a face appeared as one of the distracting items than when the face did not appear in the array. This irrelevant face effect was eliminated when the items in the arrays were inverted in Experiment 1b ruling out an explanation based on some low-level image-based properties of the faces. Experiment 2 replicated and extended the results of Experiment 1a. Irrelevant faces once again interfered with search for butterflies but, when the roles of faces and butterflies were reversed, irrelevant butterflies no longer interfered with search for faces. This suggests that the irrelevant face effect is unlikely to have been caused by the relative novelty of the faces or arises because butterflies and faces were the only animate items in the arrays. We conclude that these experiments offer evidence of a stimulus-driven capture of attention by faces.

Role for nonstructural protein 1 of severe acute respiratory syndrome coronavirus in chemokine dysregulation.

Severe acute respiratory syndrome (SARS) is an emerging infectious disease caused by a novel coronavirus. Since its associated morbidity and mortality have been postulated to be due to immune dysregulation, we investigated which of the viral proteins is responsible for chemokine overexpression. To delineate the viral and cellular factor interactions, the role of four SARS coronavirus proteins, including nonstructural protein 1 (nsp-1), nsp-5, envelope, and membrane, were examined in terms of cytokine induction. Our results showed that the SARS coronavirus nsp-1 plays an important role in CCL5, CXCL10, and CCL3 expression in human lung epithelial cells via the activation of NF-kappaB.

The impact of secondary tasks on multi-tasking in a virtual environment.

One experiment is described that examined the possible involvement of working memory in the Virtual Errands Test (McGeorge et al. (2001). Using virtual environments in the assessment of executive dysfunction. Presence, 10, 375-383), which requires participants to complete errands within a virtual environment, presented on a computer screen. Time was limited, therefore participants had to swap between tasks (multi-task) efficiently to complete the errands. Forty-two undergraduates participated, all attempting the test twice. On one of these occasions they were asked to perform a concurrent task throughout (order of single and dual-task conditions was counterbalanced). The type of secondary task was manipulated between groups. Twenty-one participants were asked to randomly generate months of the year aloud in the dual-task condition, while another 21 were asked to suppress articulation by repeating the word "December". An overall dual-task effect on the Virtual Errands Test was observed, although this was qualified by an interaction with the order of single and dual-task conditions. Analysis of the secondary task data showed a drop in performance (relative to baseline) under dual-task conditions, and that drop was greater for the random generation group than the articulatory suppression group. These data are interpreted as suggesting that the central executive and phonological loop components of working memory are implicated in this test of multi-tasking.