Normal greeble learning in a severe case of developmental prosopagnosia

A central question in cognitive neuroscience is whether mechanisms exist that are specialized for particular domains. One of the most commonly cited examples of a domain-specific competence is the human ability to recognize upright faces. However, according to a widely discussed alternative hypothesis, face recognition is instead performed by mechanisms specialized for processing any object class for which an individual has expertise. Faces, according to this domain-general hypothesis, are just one example of an expert class. Nonface object expertise has been intensively investigated using a training procedure involving an artificial stimulus class known as greebles. A key prediction of this hypothesis is that individuals with face recognition impairments will also have impairments with other categories that control subjects have expertise with. Our results show that a man with severe prosopagnosia performed normally throughout the standard greeble training procedure. These findings indicate that face recognition and greeble recognition rely on separate mechanisms.     

Quantitative analysis of indigo and indigo precursors in leaves of Isatis spp. and Polygonum tinctorium

Analysis of extracts from two woad species (Isatis tinctoria and Isatis indigotica) and Polygonum tinctorium revealed that only one indigo precursor (indican) was present in Polygonum, but two precursors were found in Isatis spp. This was done using high performance liquid chromatography (HPLC), coupled to an evaporative light scattering detector (ELSD). In Isatis spp., the indigo precursors indican and a fraction representing isatan B were identified. The proportion of indican and isatan B was different between the two Isatis spp. tested. For the first time, it was possible to quantify the precursors in woad plant species, and the results were found to be in good agreement with those made from total indigo quantification using two different spectrophotometric methods or a derivatization technique.     

Cre recombinase specificity defined by the tau locus

We generated a transgenic mouse line (tau::Cre) by targeting the Cre to the tau locus (Mapt). Based on previous reports on the expression of Tau during development, we expected the Cre recombinase to be expressed in a neuron-specific and pan-neuronal manner. However, intercrosses between the tau::Cre and the Cre-activatable reporter animals resulted in offspring with recombination either restricted to the nervous system or throughout the entire conceptus, indicating expression of Tau early in development. The percentage of neuron-specific excision was dependent on the Cre reporter used representing different Cre target sites in the mouse genome. In spite of the observed variability, our data suggest that the tau::Cre mouse line can be used for pan-neuronal recombination of floxed alleles when it is used with caution.     

KIR2DL4 is an IL-2-regulated NK cell receptor that exhibits limited expression in humans but triggers strong IFN-gamma production

Killer cell Ig-like receptor (KIR)2DL4 (2DL4, CD158d) was previously described as the only KIR expressed by every human NK cell. It is also structurally atypical among KIRs because it possesses a basic transmembrane residue, which is characteristic of many activating receptors, but also contains a cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM). We expressed epitope-tagged 2DL4 in an NK-like cell line to study receptor function. Three distinct 2DL4 cDNA clones were analyzed: one encoding the "conventional" 2DL4 with the cytoplasmic ITIM (2DL4.1) and two encoding different cytoplasmic truncated forms lacking the ITIM (2DL4.2 and 2DL4(*)). Surprisingly, one truncated receptor (2DL4.2), which is the product of a prevalent human 2DL4 allele, was not expressed on the cell surface, indicating that some individuals may lack functional 2DL4 protein expression. Conversely, both 2DL4.1 and 2DL4(*) were expressed on the cell surface and up-regulated by IL-2. Analysis of primary NK cells with anti-2DL4 mAb confirmed the lack of surface expression in a donor with the 2DL4.2 genotype. Donors with the 2DL4.1 genotype occasionally expressed receptor only on CD56(high) NK cells, although their expression was up-regulated by IL-2. Interestingly, Ab engagement of epitope-tagged 2DL4 triggered rapid and robust IFN-gamma production, but weak redirected cytotoxicity in an NK-like cell line, which was the opposite pattern to that observed upon engagement of another NK cell activating receptor, NKp44. Importantly, both 2DL4.1 and 2DL4(*) exhibited similar activation potential, indicating that the ITIM does not influence 2DL4.1 activating function. The unique activation properties of 2DL4 suggest linkage to a distinct signaling pathway.     

Elevated concentration of TNF-α induces trophoblast differentiation in mouse blastocyst outgrowths

Elevated expression of tumour necrosis factor- (TNF-) is associated with adverse pregnancy outcome. This study has examined the expression of TNF- and its receptors (TNF-Rs) by mouse blastocysts and blastocyst outgrowths from day 4 to 9.5 of pregnancy and investigated the effects of elevated TNF- on the inner cell mass (ICM) and trophoblast cells of blastocyst outgrowths. RT-PCR demonstrated TNF- mRNA expression from day 7.5 to 9.5, TNF-R1 from day 6.5 to 9.5 and TNF-R2 from day 5.5 to 7.5 of pregnancy, and in situ hybridisation revealed the trophoblast giant cells (TGCs) of the early placenta as the site of TNF- expression. Day 4 blastocysts were cultured in a physiologically high concentration of TNF- (100 ng/ml) for 72 h to the outgrowth stage and then compared to blastocysts cultured in media alone. TNF--treated blastocyst outgrowths exhibited a significant reduction in ICM cells (mean ± SD 23.90±10.42 vs 9.37±7.45, t-test, P<0.0001) with no significant change in the numbers of trophoblast cells (19.97±8.14 vs 21.73±7.79, t-test, P=0.39). Within the trophoblast cell population, the TNF--treated outgrowths exhibited a significant increase in multinucleated cells (14.10±5.53 vs 6.37±5.80, t-test, P<0.0001) and a corresponding significant decrease in mononucleated cells (5.87±3.60 vs 15.37±5.87, t-test, P<0.0001). In summary, this study describes the expression of TNF- and its receptors during the peri-implantation period in the mouse. It also reports that elevated TNF- restricts ICM proliferation in the blastocyst and changes the ratio of mononucleated to multinucleated trophoblast cells. These findings suggest a mechanism by which increased expression of TNF- during trophoblast differentiation may be detrimental to pregnancy.This work was supported by the National Health and Medical Research Council of Australia     

Cutting edge: a hypomorphic mutation in Igbeta (CD79b) in a patient with immunodeficiency and a leaky defect in B cell development

Although null mutations in Igalpha have been identified in patients with defects in B cell development, no mutations in Igbeta have been reported. We recently identified a patient with a homozygous amino acid substitution in Igbeta, a glycine to serine at codon 137, adjacent to the cysteine required for the disulfide bond between Igalpha and Igbeta. This patient has a small percentage of surface IgM(dim) B cells in the peripheral circulation (0.08% compared with 5-20% in healthy controls). Using expression vectors in 293T cells or Jurkat T cells, we show that the mutant Igbeta can form disulfide-linked complexes and bring the mu H chain to the cell surface as part of the BCR but is inefficient at both tasks. The results show that minor changes in the ability of the Igalpha/Igbeta complex to bring the BCR to the cell surface have profound effects on B cell development.     

Multi-scale modeling of a wound-healing cell migration assay

A continuum model and a discrete model are developed to capture the population-scale and cell-scale behavior in a wound-healing cell migration assay created from a scrape wound in a confluent cell monolayer. During wound closure, the cell population forms a sustained traveling wave, with close contact between cells behind the wavefront. Cells exhibit contact inhibition of migration and contact-limited proliferation. The continuum model includes the two dominant mechanisms and characteristics of cell migration and proliferation, using a cell diffusivity function that decreases with cell density and a logistic proliferative growth term. The discrete model arises naturally from the continuum model. Individual cells are simulated as continuous-time random walkers with nearest-neighbor transitions, together with a birth/death process. The migration and proliferation parameters are determined by analysing individual mice 3T3 fibroblast cell trajectories obtained during the development of a confluent cell monolayer and in a wound healing assay. The population-scale model successfully predicts the shape and speed of the traveling wave, while the discrete model is also successful in capturing the contact inhibition of migration effects.     

Coalescence of interacting cell populations

We analyse the coalescence of invasive cell populations by studying both the temporal and steady behaviour of a system of coupled reaction-diffusion equations. This problem is relevant to recent experimental observations of the dynamics of opposingly directed invasion waves of cells. Two cell types, u and v, are considered with the cell motility governed by linear or nonlinear diffusion. The cells proliferate logistically so that the long-term total cell density, u+v approaches a carrying capacity. The steady-state solutions for u and v are denoted u(s) and v(s). The steady solutions are spatially invariant and satisfy u(s)+v(s)=1. However, this expression is underdetermined so the relative proportion of each cell type u(s) and v(s) cannot be determined a priori. Various properties of this model are studied, such as how the relative proportion of u(s) and v(s) depends on the relative motility and relative proliferation rates. The model is analysed using a combination of numerical simulations and a comparison principle. This investigation unearths some novel outcomes regarding the role of overcrowding and cell death in this type of cell migration assay. These observations have relevance to experimental design and interpretation regarding the identification and parameterisation of mechanisms involved in cell invasion.