The ECM of mammalian tissues has been used as a scaffold to facilitate the repair and reconstruction of numerous tissues. Such scaffolds are prepared in many forms including sheets, powders, and hydrogels. ECM hydrogels provide advantages such as injectability, the ability to fill an irregularly shaped space, and the inherent bioactivity of native matrix. However, material properties of ECM hydrogels and the effect of these properties upon cell behavior are neither well understood nor controlled. The objective of this study was to prepare and determine the structure, mechanics, and the cell response in vitro and in vivo of ECM hydrogels prepared from decellularized porcine dermis and urinary bladder tissues. Dermal ECM hydrogels were characterized by a more dense fiber architecture and greater mechanical integrity than urinary bladder ECM hydrogels, and showed a dose dependent increase in mechanical properties with ECM concentration. In vitro, dermal ECM hydrogels supported greater C2C12 myoblast fusion, and less fibroblast infiltration and less fibroblast mediated hydrogel contraction than urinary bladder ECM hydrogels. Both hydrogels were rapidly infiltrated by host cells, primarily macrophages, when implanted in a rat abdominal wall defect. Both ECM hydrogels degraded by 35 days in vivo, but UBM hydrogels degraded more quickly, and with greater amounts of myogenesis than dermal ECM. These results show that ECM hydrogel properties can be varied and partially controlled by the scaffold tissue source, and that these properties can markedly affect cell behavior.
We investigated oculomotor anticipations in 4-month-old infants as they viewed center-occluded object trajectories. In two experiments, we examined performance in two-dimensional (2D) and three-dimensional (3D) dynamic occlusion displays and in an additional 3D condition with a smiley face as the moving target stimulus. Rates of anticipatory eye movements were not facilitated by 3D displays or by the (presumably) more salient smiley face relative to the 2D condition. However, latencies of anticipations were reduced, implying that 3D visual information may have supported formation of more robust mental representations of the moving object. Results are interpreted in a context of perceptual constraints on developing cognitive capacities during early infancy.
Infant perception; Depth perception; Visual development; Eye movements; Object knowledge
We investigated the possibility that a range of social stimuli capture the attention of 6-month-old infants when in competition with other non-face objects. Infants viewed a series of six-item arrays in which one target item was a face, body part, or animal as their eye movements were recorded. Stimulus arrays were also processed for relative salience of each item in terms of color, luminance, and amount of contour. Targets were rarely the most visually salient items in the arrays, yet infants' first looks toward all three target types were above chance, and dwell times for targets exceeded other stimulus types. Girls looked longer at faces than did boys, but there were no sex differences for other stimuli. These results are interpreted in a context of learning to discriminate between different classes of animate stimuli, perhaps in line with affordances for social interaction, and origins of sex differences in social attention.
face perception; sex differences; infant development; attention; saliency map
This is the first analysis to estimate the costs of commercially insured patients with Parkinson’s disease (PD) in the USA. Prior analyses of PD have not examined costs in patients aged under 65 years, a majority of whom are in the workforce.
Our objective was to estimate direct and indirect costs associated with PD in patients under the age of 65 years who are newly diagnosed or have evidence of advanced PD.
PD patients were selected from a commercially insured claims database (N > 12,000,000; 1999–2009); workloss data were available for a sub-sample of enrollees. Newly diagnosed patients with evidence of similar disorders were excluded. Patients with evidence of advanced PD disease, including ambulatory assistance device users (PDAAD) and institutionalized (PDINST) patients, as well as newly diagnosed PD patients, were analyzed. Each PD cohort was age-, gender- and region-matched to controls without PD. Direct (i.e. insurer payments to providers) and indirect (i.e. workloss) costs were reported in $US, year 2010 values, and were descriptively compared using Wilcoxon rank sum tests.
Patients had excess mean direct PD-related costs of $US4,072 (p < 0.001; N = 781) in the year after diagnosis. The PDAAD cohort (N = 214) had excess direct PD-related costs of $US26,467 (p < 0.001) and the PDINST cohort (N = 156) had excess direct PD-related costs of $US37,410 (p < 0.001) in the year after entering these states. Outpatient care was the most expensive cost source for newly diagnosed patients, while inpatient care was the most expensive for PDAAD and PDINST patients. Excess indirect costs were $US3,311 (p < 0.05; N = 173) in the year after initial diagnosis.
Direct costs for newly diagnosed PD patients exceeded costs for controls without PD, and increased with PD progression. Direct costs were approximately 6–7 times higher in patients with advanced PD than in matched controls. Indirect costs represented 45 % of total excess costs for newly diagnosed PD patients.
Biologic scaffolds composed of mammalian extracellular matrix (ECM) are routinely used for the repair and reconstruction of injured or missing tissues in a variety of pre-clinical and clinical applications. However, the structural and functional outcomes have varied considerably. An important variable of xenogeneic biologic scaffolds is the age of the animal from which the ECM is derived. The present study compared the in vivo host response and remodeling outcomes of biologic scaffolds composed of small intestinal submucosa (SIS)-ECM harvested from pigs that differed only in age. Results showed that there are distinct differences in the remodeling characteristics as a consequence of source animal age. Scaffolds derived from younger animals were associated with a more constructive, site appropriate, tissue remodeling response than scaffolds derived from older animals. Furthermore, the constructive remodeling response was associated with a dominant M2 macrophage response.
Age/ageing; ECM (extracellular matrix); Scaffold; Macrophage; Immune response; Mechanical properties
Tissue regeneration in response to injury in adult mammals is generally limited to select tissues. Nonmammalian species such as newts and axolotls undergo regeneration of complex tissues such as limbs and digits via recruitment and accumulation of local and circulating multipotent progenitors preprogrammed to recapitulate the missing tissue. Directed recruitment and activation of progenitor cells at a site of injury in adult mammals may alter the default wound-healing response from scar tissue toward regeneration. Bioactive molecules derived from proteolytic degradation of extracellular matrix (ECM) proteins have been shown to recruit a variety of progenitor cells in vitro and in vivo to the site of injury. The present study further characterized the population of cells accumulating at the site of injury after treatment with ECM degradation products in a well-established model of murine digit amputation. After a mid-second phalanx digit amputation in 6–8-week-old adult mice, treatment with ECM degradation products resulted in the accumulation of a heterogeneous population of cells, a subset of which expressed the transcription factor Sox2, a marker of pluripotent and adult progenitor cells. Sox2+ cells were localized lateral to the amputated P2 bone and coexpressed progenitor cell markers CD90 and Sca1. Transgenic Sox2 eGFP/+ and bone marrow chimeric mice showed that the bone marrow and blood circulation did not contribute to the Sox2+ cell population. The present study showed that, in addition to circulating progenitor cells, resident tissue-derived cells also populate at the site of injury after treatment with ECM degradation products. Although future work is necessary to determine the contribution of Sox2+ cells to functional tissue at the site of injury, recruitment and/or activation of local tissue-derived cells may be a viable approach to tissue engineering of more complex tissues in adult mammals.
Influenza A virus infection is a major global health concern causing significant mortality, morbidity, and economic loss. Antiviral chemotherapeutics that target influenza A virus are available; however, rapid emergence of drug-resistant strains has been reported. Consequently, there is a burgeoning need to identify novel anti-influenza A drugs, particularly those that target host gene products required for virus replication, to reduce the likelihood of drug resistance. In this study, a small interfering RNA (siRNA) screen was performed to identify host druggable gene targets for anti-influenza A virus therapy. The host organic anion transporter-3 gene (OAT3), a member of the SLC22 family of transporters, was validated as being required to support influenza A virus replication. Probenecid, a prototypical uricosuric agent and chemical inhibitor of organic anion transporters known to target OAT3, was shown to be effective in limiting influenza A virus infection in vitro (50% inhibitory concentration [IC50] of 5.0 × 10−5 to 5.0 × 10−4 μM; P < 0.005) and in vivo (P < 0.05). Probenecid is widely used for treatment of gout and related hyperuricemic disorders, has been extensively studied for pharmacokinetics and safety, and represents an excellent candidate for drug repositioning as a novel anti-influenza A chemotherapeutic.
Research into plant-mediated indirect interactions between arbuscular mycorrhizal (AM) fungi and insect herbivores has focussed on those between plant shoots and above-ground herbivores, despite the fact that only below-ground herbivores share the same part of the host plant as AM fungi. Using Plantago lanceolata L., we aimed to characterise how early root herbivory by the vine weevil (Otiorhynchus sulcatus F.) affected subsequent colonization by AM fungi (Glomus spp.) and determine how the two affected plant growth and defensive chemistry. We exposed four week old P. lanceolata to root herbivory and AM fungi using a 2×2 factorial design (and quantified subsequent effects on plant biomass and iridoid glycosides (IGs) concentrations. Otiorhynchus sulcatus reduced root growth by c. 64%, whereas plant growth was unaffected by AM fungi. Root herbivory reduced extent of AM fungal colonization (by c. 61%). O. sulcatus did not influence overall IG concentrations, but caused qualitative shifts in root and shoot IGs, specifically increasing the proportion of the more toxic catalpol. These changes may reflect defensive allocation in the plant against further attack. This study demonstrates that very early root herbivory during plant development can shape future patterns of AM fungal colonization and influence defensive allocation in the plant.
The elastomeric, monobloc disc prosthesis (Cadisc™-L, Ranier Technology, Cambridge, UK) aims to preserve biomechanics of an implanted spinal motion segment.
This study presents the findings of an in vitro investigation on the effect of implantation of Cadisc™-L. Compressive stiffness, flexion stiffness at 10, 20, 30 and 40 Nm and the instant-axis-of-rotation (IAR) loci are compared before and after implantation of a MC-10 mm-6° Cadisc™-L.
Fresh frozen human monosegmental lumbar spines (n = 8) were prepared, potted and tested in an environmentally controlled chamber to simulate in vivo conditions. Specimens were preconditioned by loading to 500 N for 30 min. Compressive stiffness of the specimen was determined by applying pure compression of 1 kN at 250 N/s via a loading roller positioned at the central loading axis (CLA). The roller was then offset 12.5 mm anterior of the CLA and the loading regime repeated to test specimens in flexion. Bending moments were calculated from the applied load and corresponding flexion angle. The IAR locus was tracked by a motion-tracking camera.
Compressive stiffness was reduced by 50 % (p = 0.0005), flexion stiffness was not statistically significantly reduced (40 % reduction, p > 0.05). IAR locus maintained a ‘horizontal figure of eight’ characteristic. Change in the locus width in the AP plane of 6.4 mm (p = 0.06) and height in the SI plane of 1.3 mm (p = 0.44) were not significant. The centroid was displaced 4.44 mm (p = 0.0019) and 5.44 mm (p = 0.025) at 3° and 6° flexion, respectively.
Implantation of Cadisc™-L caused a reduction in axial stiffness, but maintained disc height and flexion stiffness. IAR loci remained mobile without large displacement of the centroid from the intact spine position.
Lumbar spine; Disc replacement; Biomechanics; In vitro; Cadisc™-L
Young infants perceive an object’s trajectory as continuous across occlusion provided the temporal or spatial gap in perception is small. In three experiments involving 72 participants we investigated the effects of different forms of auditory information on 4-month-olds’ perception of trajectory continuity. Provision of dynamic auditory information about the object’s trajectory enhanced perception of trajectory continuity. However, a smaller positive effect was also obtained when the sound was continuous but provided no information about the object’s location. Finally, providing discontinuous auditory information or auditory information that was dislocated relative to vision had negative effects on trajectory perception. These results are discussed relative to the intersensory redundancy hypothesis and emphasise the need to take an intersensory approach to infant perception.
Interleukin-10 (IL-10) is an anti-inflammatory molecule that has achieved interest as a therapeutic for neuropathic pain. In this work, the potential of plasmid DNA-encoding IL-10 (pDNA-IL-10) slowly released from biodegradable microparticles to provide long-term pain relief in an animal model of neuropathic pain was investigated.
PLGA microparticles encapsulating pDNA-IL-10 were developed and assessed both in vitro and in vivo.
In vitro, pDNA containing microparticles activated macrophages, enhanced the production of nitric oxide, and increased the production of IL-10 protein relative to levels achieved with unencapsulated pDNA-IL-10. In vivo, intrathecally administered microparticles embedded in meningeal tissue, induced phagocytic cell recruitment to the cerebrospinal fluid, and relieved neuropathic pain for greater than 74 days following a single intrathecal administration, a feat not achieved with unencapsulated pDNA. Therapeutic effects of microparticle-delivered pDNA-IL-10 were blocked in the presence of IL-10-neutralizing antibody, and elevated levels of plasmid-derived IL-10 were detected in tissues for a prolonged time period post-injection (>28 days), demonstrating that therapeutic effects are dependent on IL-10 protein production.
These studies demonstrate that microparticle encapsulation significantly enhances the potency of intrathecally administered pDNA, which may be extended to treat other disorders that require intrathecal gene therapy.
interleukin-10; microparticle; pDNA; PLGA; neuropathic pain
Much research in the past two decades has documented infants’ and adults’ ability to extract statistical regularities from auditory input. Importantly, recent research has extended these findings to the visual domain, demonstrating learners’ sensitivity to statistical patterns within visual arrays and sequences of shapes. In this review we discuss both auditory and visual statistical learning to elucidate both the generality of and constraints on statistical learning. The review first outlines the major findings of the statistical learning literature with infants, followed by discussion of statistical learning across domains, modalities, and development. The second part of this review considers constraints on statistical learning. The discussion focuses on two categories of constraint: constraints on the types of input over which statistical learning operates and constraints based on the state of the learner. The review concludes with a discussion of possible mechanisms underlying statistical learning.
infants; auditory statistical learning; visual statistical learning; language acquistion; learning constraints; statistical learning mechanisms
Biologic scaffolds composed of extracellular matrix (ECM) have been used successfully in preclinical models and humans for constructive remodeling of functional, site-appropriate tissue after injury. The mechanisms underlying ECM-mediated constructive remodeling are not completely understood, but scaffold degradation and site-directed recruitment of progenitor cells are thought to play critical roles. Previous studies have identified a cryptic peptide derived from the C-terminal telopeptide of collagen IIIα that has chemotactic activity for progenitor cells. The present study characterized the osteogenic activity of the same peptide in vitro and in vivo in an adult murine model of digit amputation. The present study showed that the cryptic peptide increased calcium deposition, alkaline phosphatase activity, and osteogenic gene expression in human perivascular stem cells in vitro. Treatment with the cryptic peptide in a murine model of mid-second phalanx digit amputation led to the formation of a bone nodule at the site of amputation. In addition to potential therapeutic implications for the treatment of bone injuries and facilitation of reconstructive surgical procedures, cryptic peptides with the ability to alter stem cell recruitment and differentiation at a site of injury may serve as powerful new tools for influencing stem cell fate in the local injury microenvironment.
Biologic scaffolds composed of extracellular matrix (ECM) have been used successfully in preclinical models and humans for constructive remodeling of functional, site-appropriate tissue after injury. The mechanisms underlying ECM-mediated constructive remodeling are not completely understood, but scaffold degradation and site-directed recruitment of both differentiated and progenitor cells are thought to play critical roles. Previous studies have shown that degradation products of ECM scaffolds can recruit a population of progenitor cells both in vitro and in vivo. The present study identified a single cryptic peptide derived from the α subunit of the collagen III molecule that is chemotactic for a well-characterized perivascular stem cell in vitro and causes the site-directed accumulation of progenitor cells in vivo. The oligopeptide was additionally chemotactic for human cortical neural stem cells, rat adipocyte stem cells, C2C12 myoblast cells, and rat Schwann cells in vitro. In an adult murine model of digit amputation, treatment with this peptide after mid-second phalanx amputation resulted in a greater number of Sox2+ and Sca1+,Lin− cells at the site of injury compared to controls. Since progenitor cell activation and recruitment are key prerequisites for epimorphic regeneration in adult mammalian tissues, endogenous site-directed recruitment of such cells has the potential to alter the default wound healing response from scar tissue toward regeneration.
Word-learning likely involves a multiplicity of components, some domain-general, others domain-specific. Against the background of recent studies that suggest that word-learning is domain-specific, we investigated the associative component of word-learning. Seven- and 14-month-old infants viewed a pair of events in which a monkey or a truck moved back and forth, accompanied by a sung syllable or a tone, matched for pitch. Following habituation, infants were presented with displays in which the visual-auditory pairings were preserved or switched, and looked longer at the “switch” events when exposure time was sufficient to learn the intermodal association. At 7 months, performance on speech and tones conditions was statistically identical; at 14 months, infants had begun to favor speech. Thus, the associative component of word-learning does not appear (in contrast to rule-learning, Marcus et al., 2007) to initially privilege speech.
word-learning; association; cognitive development
The Notch signaling pathway is an intercellular communication network vital to metazoan development. Notch activation leads to the nuclear localization of the intracellular portion (NICD) of the Notch receptor. Once in the nucleus, NICD binds the transcription factor CSL through a bivalent interaction involving the high-affinity RAM region and the lower affinity ANK domain, converting CSL from a transcriptionally-repressed to an active state. This interaction is believed to directly displace co-repressor proteins from CSL and recruit co-activator proteins. Here we investigate the consequences of this bivalent organization in converting CSL from the repressed to active form. One proposed function of RAM is to promote the weak ANK:CSL interaction; thus, fusion of CSL-ANK should bypass this function of RAM. We find that a CSL-ANK fusion protein is transcriptionally active in reporter assays, but that the addition of RAM in trans further increases transcriptional activity, suggesting another role of RAM in activation. A single F235L point substitution, which disrupts co-repressor binding to CSL, renders the CSL-ANK fusion fully active and refractory to further stimulation by RAM in trans. These results suggest that in the context of a mammalian CSL-ANK fusion protein, the main role of RAM is to displace co-repressor proteins from CSL.
Infants appear to learn abstract rule-like regularities (e.g., la la da follows an AAB pattern) more easily from speech than from a variety of other auditory and visual stimuli (Marcus et al., 2007). We test if that facilitation reflects a specialization to learn from speech alone, or from modality-independent communicative stimuli more generally, by measuring 7.5-month-old infants’ ability to learn abstract rules from sign language-like gestures. Whereas infants appear to easily learn many different rules from speech, we found that with sign-like stimuli, and under circumstances comparable to those of Marcus et al. (1999), hearing infants were able to learn an ABB rule, but not an AAB rule. This is consistent with results of studies that demonstrate lower levels of infant rule learning from a variety of other non-speech stimuli, and we discuss implications for accounts of speech-facilitation.
From birth, infants detect associations between the locations of static visual objects and sounds they emit, but there is limited evidence regarding their sensitivity to the dynamic equivalent when a sound-emitting object moves. In four experiments involving 36 2-month-olds, 48 5-month-olds and 48 8-month-olds, we investigated infants’ ability to process this form of spatial co-location. Whereas there was no evidence of spontaneous sensitivity, all age groups detected a dynamic co-location during habituation, and looked longer at test trials in which sound and sight were dislocated. Only 2-month-olds showed clear sensitivity to the dislocation relation, although 8-month-olds did so following additional habituation. These results are discussed relative to the intersensory redundancy hypothesis and work suggesting increasing specificity in processing with age.
Live-attenuated influenza vaccine (LAIV) delivered by large droplet intranasal spray is efficacious against infection. However, many of the large droplets are trapped in the external nares and do not reach the target nasal airway tissues. Smaller droplets might provide better distribution yielding similar protection with lower doses. We evaluated 20 and 30 micron aerosol delivery of influenza virus in mice. A 15 second aerosol exposure optimally protected against homologous and heterologous influenza infection and induced a robust immune response. These results demonstrate the feasibility of nasal vaccination using aerosolized particles, providing a strategy to improve vaccine efficacy and delivery.
influenza; vaccination; aerosol
The visual world of adults consists of objects at various distances, partly occluding one another, substantial and stable across space and time. The visual world of young infants, in contrast, is often fragmented and unstable, consisting not of coherent objects but rather surfaces that move in unpredictable ways. Evidence from computational modeling and from experiments with human infants highlights three kinds of learning that contribute to infants' knowledge of the visual world: learning via association, learning via active assembly, and learning via visual-manual exploration. Infants acquire knowledge by observing objects move in and out of sight, forming associations of these different views. In addition, the infant's own self-produced behavior—oculomotor patterns and manual experience, in particular—are important means by which infants discover and construct their visual world.
Visual development; cognitive development; models of development; object perception; infants; learning
In everyday environments, objects frequently go out of sight as they move and our view of them becomes obstructed by nearer objects, yet we perceive these objects as continuous and enduring entities. Here, we used functional MRI with an attentive tracking paradigm to clarify the nature of perceptual and cognitive mechanisms subserving this ability to fill in the gaps in perception of dynamic object occlusion. Imaging data revealed distinct regions of cortex showing increased activity during periods of occlusion relative to full visibility. These regions may support active maintenance of a representation of the target’s spatiotemporal properties ensuring that the object is perceived as a persisting entity when occluded. Our findings may shed light on the neural substrates involved in object tracking that give rise to the phenomenon of object permanence.
Opportunistic CNS-infection represent a major threat to patients after organ transplantation due to the need for ongoing immunosuppression and belatacept is a novel CTL4A inhibitor, which is increasingly used for patients following cadaveric kidney transplantation. Among the CNS infections, intracranial Aspergillus is a particular challenge and poses difficulties for its insidious onset, a timely and accurate diagnosis, and its management due to high mortality rates. To this end we want to illustrate the management of this scenario as encountered in a 71-year-old female patient, who was admitted into our institution in June 2007 with speech difficulties and gait instability 1.5 years after cadaveric kidney transplantation. On imaging, both a mediastinal and left frontal mass were found. Radiographically guided sampling of the mediastinal mass and a stereotactic biopsy of the left frontal brain lesion revealed Aspergillus fumigatus. With modification of immunosuppression and directed antifungal therapy there was complete resolution of the chest lesion; the brain lesion initially responded well but later progressed in size. Surgical intervention via a left fronto-temporal craniotomy with intraoperative image guidance was performed for a gross total resection of the lesion. Twenty-four months from resection, she remains on voriconazole with no evidence of recurrence and complete neurologic recovery and preserved renal function.
Belatacept; CNS aspergillosis; immunosuppression; neurosurgery; renal transplant; voriconazole
The potential association of variant surface glycoprotein (VSG) gene expression with clonal expression of virulence in African trypanosomes was addressed. Two populations of clonally related trypanosomes, which differ dramatically in virulence for the infected host, but display the same apparent VSG surface coat phenotype, were characterized with respect to the VSG genes expressed as well as the chromosome telomeric expression sites (ES) utilized for VSG gene transcription. The VSG gene sequences expressed by clones LouTat 1 and LouTat 1A of Trypanosoma brucei rhodesiense were identical, and gene expression in both clones occurred precisely by the same gene conversion events (duplication and transposition), which generated an expression-linked copy (ELC) of the VSG gene. The ELC was present on the same genomic restriction fragments in both populations and resided in the telomere of a 330-kb chromosome; a single basic copy of the LouTat 1/1A VSG gene, present in all variants of the LouTat 1 serodeme, was located at an internal site of a 1.5-Mb chromosome. Restriction endonuclease mapping of the ES telomere revealed that the VSG ELC of clones LouTat 1 and 1A resides in the same site. Therefore, these findings provide evidence that the VSG gene ES and, potentially, any cotranscribed ES-associated genes do not play a role in the clonal regulation of virulence because trypanosome clones LouTat 1 and 1A, which differ markedly in their virulence properties, both express identical VSG genes from the same chromosome telomeric ES.
A prototype, real-time reverse-transcription PCR assay, based on MultiCode-RTx technology, quantifying hepatitis C virus (HCV) RNA by targeting the HCV 3′ untranslated region demonstrated linearity over 7 logs, with a good correlation between the quantitative results of this assay and the results of two commercially available comparator assays for 466 clinical specimens comprising all six HCV genotypes.
In simple tests of preference, infants as young as newborns prefer faces and face-like stimuli over distractors. Little is known, however, about the development of attention to faces in complex scenes. We recorded eye-movements of 3-, 6-, and 9-month-old infants and adults during free-viewing of clips from A Charlie Brown Christmas (an animated film). The tendency to look at faces increased with age. Using novel computational tools, we found that 3-month-olds were less consistent (across individuals) in where they looked than were older infants. Moreover, younger infants’ fixations were best predicted by low-level image salience, rather than the locations of faces. Between 3 and 9 months of age, infants gradually focus their attention on faces. We discuss several possible interpretations of this shift in terms of social development, cross-modal integration, and attentional/executive control.