Restricting microbial exposure in early life negates the immune benefits associated with gut colonization in environments of high microbial diversity

Background: Acquisition of the intestinal microbiota in early life corresponds with the development of the mucosal immune system. Recent work on caesarean-delivered infants revealed that early microbial composition is influenced by birthing method and environment. Furthermore, we have confirmed that early-life environment strongly influences both the adult gut microbiota and development of the gut immune system. Here, we address the impact of limiting microbial exposure after initial colonization on the development of adult gut immunity. Methodology/Principal Findings: Piglets were born in indoor or outdoor rearing units, allowing natural colonization in the immediate period after birth, prior to transfer to high-health status isolators. Strikingly, gut closure and morphological development were strongly affected by isolator-rearing, independent of indoor or outdoor origins of piglets. Isolator-reared animals showed extensive vacuolation and disorganization of the gut epithelium, inferring that normal gut closure requires maturation factors present in maternal milk. Although morphological maturation and gut closure were delayed in isolatorreared animals, these hard-wired events occurred later in development. Type I IFN, IL-22, IL-23 and Th17 pathways were increased in indoor-isolator compared to outdoor-isolator animals during early life, indicating greater immune activation in pigs originating from indoor environments reflecting differences in the early microbiota. This difference was less apparent later in development due to enhanced immune activation and convergence of the microbiota in all isolator-reared animals. This correlated with elevation of Type I IFN pathways in both groups, although T cell pathways were still more affected in indoor-reared animals. Conclusions/Significance: Environmental factors, in particular microbial exposure, influence expression of a large number of immune-related genes. However, the homeostatic effects of microbial colonization in outdoor environments require sustained microbial exposure throughout development. Gut development in high-hygiene environments negatively impacts on normal succession of the gut microbiota and promotes innate immune activation which may impair immune homeostasis.

Interaction between grammatical categories and cognition in bilinguals: The role of proficiency, cultural immersion, and language of instruction

Previous studies have demonstrated that there is a tight link between grammatical concepts and cognitive preferences in monolingual speakers (Lucy 1992, Lucy & Gaskins 2003, Imai & Gentner 1997, Imai & Mazuka 2003). Recent research has also shown that bilinguals with languages that differ in their concepts may shift their cognitive preferences as a function of their proficiency (Athanasopoulos, 2006) or cultural immersion (Cook, Bassetti, Kasai, Sasaki, & Takahashi, 2006). The current short paper assesses the relative impact of each of these variables, and furthermore asks whether bilinguals alternate between two distinct cognitive representations of language-specific concepts depending on the language used in the experiment. Results from an object classification task showed that Japanese–English bilinguals shifted their behaviour towards the second language (L2) pattern primarily as a function of their L2 proficiency, while cultural immersion and language of instruction played a minimal role. These findings suggest that acquisition of novel grammatical categories leads to cognitive restructuring in the bilingual mind and have implications for the relationship between language and cognitive processing.

Within-host evolution of Staphylococcus aureus during asymptomatic carriage

Background Staphylococcus aureus is a major cause of healthcare associated mortality, but like many important bacterial pathogens, it is a common constituent of the normal human body flora. Around a third of healthy adults are carriers. Recent evidence suggests that evolution of S. aureus during nasal carriage may be associated with progression to invasive disease. However, a more detailed understanding of within-host evolution under natural conditions is required to appreciate the evolutionary and mechanistic reasons why commensal bacteria such as S. aureus cause disease. Therefore we examined in detail the evolutionary dynamics of normal, asymptomatic carriage. Sequencing a total of 131 genomes across 13 singly colonized hosts using the Illumina platform, we investigated diversity, selection, population dynamics and transmission during the short-term evolution of S. aureus. Principal Findings We characterized the processes by which the raw material for evolution is generated: micro-mutation (point mutation and small insertions/deletions), macro-mutation (large insertions/deletions) and the loss or acquisition of mobile elements (plasmids and bacteriophages). Through an analysis of synonymous, non-synonymous and intergenic mutations we discovered a fitness landscape dominated by purifying selection, with rare examples of adaptive change in genes encoding surface-anchored proteins and an enterotoxin. We found evidence for dramatic, hundred-fold fluctuations in the size of the within-host population over time, which we related to the cycle of colonization and clearance. Using a newly-developed population genetics approach to detect recent transmission among hosts, we revealed evidence for recent transmission between some of our subjects, including a husband and wife both carrying populations of methicillin-resistant S. aureus (MRSA). Significance This investigation begins to paint a picture of the within-host evolution of an important bacterial pathogen during its prevailing natural state, asymptomatic carriage. These results also have wider significance as a benchmark for future systematic studies of evolution during invasive S. aureus disease.

Individual differences and the role of the L1 in L2 processing: an ERP investigation

This study used ERP (event-related potentials) to examine both the role of the L1 and the role of individual differences in the processing of agreement violations. Theories of L2 acquisition differ with regard to whether or not native-like acquisition of L2 features is possible (Schwartz and Sprouse, 1994, 1996; Tsimpli and Mastropavlou, 2007), and the results of previous ERP studies are inconsistent when it comes to whether or not native-like processing is observed in response to L2 agreement violations (e.g., Sabourin, 2003; Tokowicz and MacWhinney, 2005). Furthermore, studies of learners in early stages of L2 acquisition have found variability in the emergence of native-like responses (e.g., McLaughlin et al., 2010; Tanner et al., 2009), but sources of variability have not been investigated. The current study examines responses to gender and number agreement violations in English-speaking learners of Spanish (n=24). Stimuli targeted agreement in three conditions: subject-verb agreement (el barco flota/*flotan), which is similar in Spanish and English; number agreement on adjectival predicates (la isla rocosa/*rocosas), a context in which agreement is not instantiated in English; and gender agreement on adjectival predicates (la isla rocosa/*rocoso), which is unique to Spanish. Grammaticality judgments and ERP responses were also tested for correlations with aptitude scores on the Modern Languages Aptitude Test (MLAT; Carroll and Sapon, 1959) and the Raven Advanced Progressive Matrices (Raven, 1965). Results are in line with theories that claim native-like processing is acquirable, since learners demonstrated similar ERP responses to a control group of native Spanish-speakers (n=8) with regard to all three agreement types. Additionally, the MLAT (but not the Raven) was significantly correlated with sensitivity to number violations, both in terms of grammaticality judgments and ERP amplitudes, indicating a role for verbal but not nonverbal aptitude in L2 processing.

Children's misconceptions and the teaching of early years science: a case study

The aim of this paper is to examine how teachers’ awareness of children’s misconceptions can affect children’s acquisition of scientific concepts. In other words, this paper is aimed at examining whether teaching is altered when teachers are aware of pupils’ misconceptions of a specific science concept. This paper details a case study focused on two kindergarten classes of five year-old children and their teachers and took place in Cyprus. Two lessons were observed and three children from each class were interviewed. Through the analysis of children’s responses it was possible to identify specific misconceptions related to the concept of rain. The results indicate that it is very important for teachers to be aware of what misconceptions children have, because this can help them plan lessons for children to overcome their misconceptions. It seems that it is more likely for children to overcome their misconceptions when teachers take these misconceptions into account as they plan and teach science lessons.

Highly efficient neural differentiation of human somatic stem cells, isolated by minimally invasive periodontal surgery

Neural stem cells (NSCs) are potential sources for cell therapy of neurodegenerative diseases and for drug screening. Despite their potential benefits, ethical and practical considerations limit the application of NSCs derived from human embryonic stem cells (ES) or adult brain tissue. Thus, alternative sources are required to satisfy the criteria of ready accessibility, rapid expansion in chemically defined media and reliable induction to a neuronal fate. We isolated somatic stem cells from the human periodontium that were collected during minimally invasive periodontal access flap surgery as part of guided tissue regeneration therapy. These cells could be propagated as neurospheres in serum-free medium, which underscores their cranial neural crest cell origin. Culture in the presence of epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2) under serum-free conditions resulted in large numbers of nestin-positive/Sox-2-positive NSCs. These periodontium-derived (pd) NSCs are highly proliferative and migrate in response to chemokines that have been described as inducing NSC migration. We used immunocytochemical techniques and RT-PCR analysis to assess neural differentiation after treatment of the expanded cells with a novel induction medium. Adherence to substrate, growth factor deprivation, and retinoic acid treatment led to the acquisition of neuronal morphology and stable expression of markers of neuronal differentiation by more than 90% of the cells. Thus, our novel method might provide nearly limitless numbers of neuronal precursors from a readily accessible autologous adult human source, which could be used as a platform for further experimental studies and has potential therapeutic implications.

Alternative generation of CNS neural stem cells and PNS derivatives from neural crest-derived peripheral stem cells

Neural crest-derived stem cells (NCSCs) from the embryonic peripheral nervous system (PNS) can be reprogrammed in neurosphere (NS) culture to rNCSCs that produce central nervous system (CNS) progeny, including myelinating oligodendrocytes. Using global gene expression analysis we now demonstrate that rNCSCs completely lose their previous PNS characteristics and acquire the identity of neural stem cells derived from embryonic spinal cord. Reprogramming proceeds rapidly and results in a homogenous population of Olig2-, Sox3-, and Lex-positive CNS stem cells. Low-level expression of pluripotency inducing genes Oct4, Nanog, and Klf4 argues against a transient pluripotent state during reprogramming. The acquisition of CNS properties is prevented in the presence of BMP4 (BMP NCSCs) as shown by marker gene expression and the potential to produce PNS neurons and glia. In addition, genes characteristic for mesenchymal and perivascular progenitors are expressed, which suggests that BMP NCSCs are directed toward a pericyte progenitor/mesenchymal stem cell (MSC) fate. Adult NCSCs from mouse palate, an easily accessible source of adult NCSCs, display strikingly similar properties. They do not generate cells with CNS characteristics but lose the neural crest markers Sox10 and p75 and produce MSC-like cells. These findings show that embryonic NCSCs acquire a full CNS identity in NS culture. In contrast, MSC-like cells are generated from BMP NCSCs and pNCSCs, which reveals that postmigratory NCSCs are a source for MSC-like cells up to the adult stage.

Neonatal environment exerts a sustained influence on the development of the intestinal microbiota and metabolic phenotype

The postnatal environment, including factors such as weaning and acquisition of the gut microbiota, has been causally linked to the development of later immunological diseases such as allergy and autoimmunity, and has also been associated with a predisposition to metabolic disorders. We show that the very early-life environment influences the development of both the gut microbiota and host metabolic phenotype in a porcine model of human infants. Farmpiglets were nursed by their mothers for 1 day, before removal to highly controlled, individual isolators where they received formula milk until weaning at 21 days. The experiment was repeated, to create two batches, which differed only in minor environmental fluctuations during the first day. At day 1 after birth, metabolic profiling of serum by 1H nuclear magnetic resonance spectroscopy demonstrated significant, systemic, inter-batch variation which persisted until weaning. However, the urinary metabolic profiles demonstrated that significant inter-batch effects on 3-hydroxyisovalerate, trimethylamine-N-oxide and mannitol persisted beyond weaning to at least 35 days. Batch effects were linked to significant differences in the composition of colonic microbiota at 35 days, determined by 16 S pyrosequencing. Different weaning diets modulated both the microbiota and metabolic phenotype independently of the persistent batch effects. We demonstrate that the environment during the first day of life influences development of the microbiota and metabolic phenotype and thus should be taken into account when interrogating experimental outcomes. In addition, we suggest that intervention at this early time could provide ‘metabolic rescue’ for at-risk infants who have undergone aberrant patterns of initial intestinal colonisation.

The neurogenic potential of astrocytes is regulated by inflammatory signals

Although the adult brain contains neural stem cells (NSCs) that generate new neurons throughout life, these astrocyte-like populations are restricted to two discrete niches. Despite their terminally differentiated phenotype, adult parenchymal astrocytes can re-acquire NSC-like characteristics following injury, and as such, these 'reactive' astrocytes offer an alternative source of cells for central nervous system (CNS) repair following injury or disease. At present, the mechanisms that regulate the potential of different types of astrocytes are poorly understood. We used in vitro and ex vivo astrocytes to identify candidate pathways important for regulation of astrocyte potential. Using in vitro neural progenitor cell (NPC)-derived astrocytes, we found that exposure of more lineage-restricted astrocytes to either tumor necrosis factor alpha (TNF-α) (via nuclear factor-κB (NFκB)) or the bone morphogenetic protein (BMP) inhibitor, noggin, led to re-acquisition of NPC properties accompanied by transcriptomic and epigenetic changes consistent with a more neurogenic, NPC-like state. Comparative analyses of microarray data from in vitro-derived and ex vivo postnatal parenchymal astrocytes identified several common pathways and upstream regulators associated with inflammation (including transforming growth factor (TGF)-β1 and peroxisome proliferator-activated receptor gamma (PPARγ)) and cell cycle control (including TP53) as candidate regulators of astrocyte phenotype and potential. We propose that inflammatory signalling may control the normal, progressive restriction in potential of differentiating astrocytes as well as under reactive conditions and represent future targets for therapies to harness the latent neurogenic capacity of parenchymal astrocytes.

Bound Variable, Split Antecedent and Ellipsis Interpretations in L2 Portuguese: Implications for L2 Acquisition Theories.

Recently, in light of minimalist assumptions, some partial UG accessibility accounts to adult second language acquisition have made a distinction between the post-critical period ability to acquire new features based on their LF-interpretability (i.e. interpretable vs. uninterpretable features) (HAWKINS, 2005; HAWKINS; HATTORI, 2006; TSIMPLI; MASTROPAVLOU, 2007; TSIMPLI; DIMITRAKOPOULOU, 2007). The Interpretability Hypothesis (TSIMPLI; MASTROPAVLOU, 2007; TSIMPLI; DIMITRAKOPOULOU, 2007) claims that only uninterpretable features suffer a post-critical period failure and, therefore, cannot be acquired. Conversely, Full Access approaches claim that L2 learners have full access to UG’s entire inventory of features, and that L1/L2 differences obtain outside the narrow syntax. The phenomenon studied herein, adult acquisition of the Overt Pronoun Constraint (OPC) (MONTALBETTI, 1984) and inflected infinitives in nonnative Portuguese, challenges the Interpretability hypothesis insofar as it makes the wrong predictions for what is observed. The present data demonstrate that advanced learners of L2 Portuguese acquire the OPC and the syntax and semantics of inflected infinitives with native-like accuracy. Since inflected infinitives require the acquisition of new uninterpretable φ-features, the present data provide evidence in contra Tsimpli and colleagues’ Interpretability Hypothesis.

How Pragmatically Odd! Interface Delays and Pronominal Subject Distribution in the L2 Spanish of English Natives

Sorace (2000, 2005) has claimed that while L2 learners can easily acquire properties of L2 narrow syntax they have significant difficulty with regard to interpretation and the discourse distribution of related properties, resulting in so-called residual optionality. However, there is no consensus as to what this difficulty indicates. Is it related to an insurmountable grammatical representational deficit (in the sense of representation deficit approaches; e.g. Beck 1998, Franceschina 2001, Hawkins 2005), is it due to cross-linguistic interference, or is it just a delay due to a greater complexity involved in the acquisition of interface-conditioned properties? In this article, I explore the L2 distribution of null and overt subject pronouns of English speaking learners of L2 Spanish. While intermediate learners clearly have knowledge of the syntax of Spanish null subjects, they do not have target-like pragmatic knowledge of their distribution with overt subjects. The present data demonstrate, however, that this difficulty is overcome at highly advanced stages of L2 development, thus suggesting that properties at the syntax-pragmatics interface are not destined for inevitable fossilization.

The Syntax of Pronominal Subjects in L2 Spanish: Comparing Two L2 Populations with Different Exposure

The purpose of this article is two-fold. First, via a critical review of available studies on the adult L2 resetting of the Null-Subject Parameter (NSP) and in light of a typologically wide sampling of languages, we conclude that the NSP cluster is much narrower in scope than is reflected in the design and discussion of most L2 studies. Secondly, we present original research on the L2 resetting of the NSP by two groups of adult English intermediate learners of L2 Spanish: a study-abroad group and a class-room only group. We seek to quantify the extent to which study-abroad experience, that is, increased exposure to native input, is beneficial specifically as it relates to the acquisition of new functional features needed for parameter re-setting (cf. Isabelli 2004). Despite the observable and clandestine linguistic benefits to study-abroad, our data suggest that for the resetting of the NSP, at least, such exposure to native input is not particularly gainful.