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Comparative genomics of the early diverging metazoan lineages and of their unicellular sister-groups opens new window to reconstructing the genetic changes which preceded or accompanied the evolution of multicellular body plans. A recent analysis found that the genome of the nerve-less sponges encodes the homologues of most vertebrate post-synaptic proteins. In vertebrate excitatory synapses, these proteins assemble to form the post-synaptic density, a complex molecular platform linking membrane receptors, components of their signalling pathways, and the cytoskeleton. Newly available genomes from Monosiga brevicollis (a member of Choanoflagellata, the closest unicellular relatives of animals) and Trichoplax adhaerens (a member of Placozoa: besides sponges, the only nerve-less metazoans) offer an opportunity to refine our understanding of post-synaptic protein evolution.
Searches for orthologous proteins and reconstruction of gene gains/losses based on the taxon phylogeny indicate that post-synaptic proteins originated in two main steps. The backbone scaffold proteins (Shank, Homer, DLG) and some of their partners were acquired in a unicellular ancestor of choanoflagellates and metazoans. A substantial additional set appeared in an exclusive ancestor of the Metazoa. The placozoan genome contains most post-synaptic genes but lacks some of them. Notably, the master-scaffold protein Shank might have been lost secondarily in the placozoan lineage.
The time of origination of most post-synaptic proteins was not concomitant with the acquisition of synapses or neural-like cells. The backbone of the scaffold emerged in a unicellular context and was probably not involved in cell-cell communication. Based on the reconstructed protein composition and potential interactions, its ancestral function could have been to link calcium signalling and cytoskeleton regulation. The complex later became integrated into the evolving synapse through the addition of novel functionalities.
Many of the complex traits that distinguish the organisation and biology of multicellular animals originated during the early phase of their evolutionary history. Among these, the emergence of the neuro-sensory system represents a particularly fascinating conundrum, because the biological support of animal perception and behaviour is extraordinarily complex at the molecular, cellular and anatomical scales, and without equivalent in the living world. A promising path to understanding the genetic changes that shaped the body plan of multicellular animals has been recently opened by comparative analyses of genomes from the early-diverging animal lineages and the closest unicellular relatives of metazoans (e.g. [1-10])
The recent characterisation of a nearly complete set of post-synaptic protein orthologues in the genome of the sponge Amphimedon queenslandica [2] represents a successful illustration of this approach. Excitatory synapses in the vertebrate central nervous system are characterised by an electron-dense structure called the post-synaptic density (PSD) below the post-synaptic membrane. Proteomic investigations have identified about one thousand proteins in this complex [11-14]. Some of these proteins (e.g. Shank, DLG, Homer, GRIP) have pivotal roles in building a scaffold for the anchorage of the other PSD components, for instance membrane receptors for neurotransmitters, cation channels, members of intracellular signalling pathways, and cytoskeleton or cytoskeleton regulation proteins [15-18]. This complex sub-membrane plateform thereby connects cell surface receptors with components of signal transduction pathways and with the cytoskeleton machinery. The PSD is also involved in synaptogenesis, dendritic spine formation and synaptic plasticity [17,19,20]. The three-dimensional architecture of the PSD is determined by multiple protein-protein interactions through binding domains (e.g. [17,18,21]). For example, the vertebrate Shank protein, the master scaffold protein of the PSD [18], contains five interacting domains (from N-ter to C-ter: ankyrin repeats, SH3 domain, PDZ domain, proline-rich region, SAM domain) involved in a huge variety of homomeric and heteromeric interactions. Among PSD scaffold proteins, PDZ (for Post-synaptic density protein 95/Drosophila disc large tumor suppressor/Zo-1) is the most common type of binding domain, with many proteins containing multiple PDZ domains (e.g. three in the DLG protein; seven in GRIP).
Not only were most PSD proteins identified in the sponge genome (whereas most are absent in fungal or plant genomes), but moreover these proteins essentially contain the same interacting domains arranged into the same architectures as in their vertebrate counterparts [2]. Intriguingly, several of the sponge PSD-like genes (DLG, GKAP, GRIP, HOMER, CRIPT) were found co-expressed in a particular non-ciliated epidermal cell type of the Amphimedon larva, the globular cell (erroneously called "flask cells" in [2], see [22]). These observations are consistent with the existence in sponges of an assembled structure similar to the vertebrate PSD, despite absence of recognisable nerve cells and synapses.
The PSD protein complex would therefore have originated before the acquisition of synapses in a nerve-less ancestor, at least under the conventional view that sponges branched off earlier than animals with a nervous system in the phylogeny (e.g. [23-27]). Moreover, since the sponge globular cells express Notch/Delta homologues and a proneural-like bHLH transcription factor [22] in addition to PSD genes, they might represent putative sensory "proto-neural" cells responding to environmental stimuli [22] and a possible "starting point" for the evolution of neural cells [2]. Some recent molecular phylogenies [28-31] have pointed to more complicated evolutionary scenarios (e.g. secondary loss of nerve cells and synapses in sponges) by challenging the monophyly of animals with a nervous system (Cnidaria, Ctenophora and Bilateria, classically grouped under the Eumetazoa) and the early divergence of sponges (discussed in [32,33]). However, a recent phylogenomic study with increased species sampling of non-bilaterian lineages and sophisticated model of sequence evolution [34] lent support to the conventional phylogeny and thereby to a single acquisition of nerve cells and synapses in a eumetazoan ancestor.
Since the publication of [2], additional annotated complete genomes have become available. The present study aims at improving our understanding of the emergence of the PSD protein complex by focusing on sequence data from Monosiga brevicollis and Trichoplax adhaerens, two organisms of particular interest with respect to early metazoan evolution. Monosiga is a member of choanoflagellates, the unicellular sister-group of the Metazoa [1,8]. Previous studies have suggested (without rigorous analyses) the existence of some PSD protein orthologues in the Monosiga genome [2], but these have been considered "very few"[10], supporting the idea that the PSD scaffold is essentially an innovation of the metazoan lineage. However, this remains to be tested rigorously through systematic analyses of choanoflagellate orthologues of the post-synaptic genes. Trichoplax (phylum Placozoa) is a nerve-less animal whose flat and asymmetric body comprises two epithelial cell layers enclosing a loose formation of interconnected fibre cells sandwiched in between [35]. The phylogenetic placement of placozoans remains highly contentious (e.g. [28,34,36]). Analyses of the Trichoplax genome have revealed a degree of genetic complexity that was totally unexpected given its morpho-anatomical simplicity [36]. As the only non-sponge animal lacking a nervous system, Trichoplax is clearly of pivotal importance for understanding the origin of neural cells and synapses.
The gene set investigated in the present study comprises the main scaffold proteins of the PSD, and their more significant interacting partners as recognised in the recent literature (e.g. [15-18]). It is essentially the gene set analysed in [2]. Similarity searches for PSD proteins were conducted against the genomes of M. brevicollis, T. adhaerens, several non-metazoan eukaryotes (as outgroups) and a represensative sampling of metazoan species. Our analyses provide evidence for the presence of a substantial subset of functionally important scaffold PSD proteins in the common ancestor of choanoflagellates and metazoans, and for secondary loss of the master scaffold protein Shank in placozoans.
Data collection
The protein set examined in this study is listed in Fig. 1 and Additional file 1. It is the same as in ref [2], except that we discarded GABA-B receptor, Tamalin and LIM kinase (because we were unable to find evidence from the literature for physical interaction of these proteins with the PSD scaffold), as well as Kir and Mint (the latter is a presynaptic adaptor protein [37] ; the former is a large family of inwardly rectifying potassium channels which is not particularly associated with the PSD, see [38]). In addition to the full protein alignments, we built a separate alignment for PDZ domains (as in ref [2] but with an extended gene sampling) and constructed specific alignments for the SAM and SH3 domains of Shank. The starting data set [2] comprised sequences from Saccharomyces cerevisiae (Ascomycota, Fungi), plants (Arabidopsis thaliana and Oryza sativa), Dictyostelium discoideum (Ameobozoa), Amphimedon queenslandica (Demospongiae, Porifera), Nematostella vectensis (Anthozoa, Cnidaria), Drosophila melanogaster (Hexapoda, Arthropoda) and Homo sapiens. These files were updated by TBLASTN searches (using proteins from Drosophila melanogaster as input sequences) on the most up-to-date collections of predicted transcripts from plants (Arabidopsis thaliana and Oryza sativa), Dictyostelium discoideum (Ameobozoa), fungi (Batrachochytridium dendrobatidis, Saccharomyces cerevisiae, Neurospora crassa, Ustilago maydis, Rhizopus oryzae), Monosiga brevicolis (Choanoflagellata), Trichoplax adhaerens (Placozoa), cnidarians (Hydra magnipapillata, Hydrozoa, and Nematostella vectensis, Anthozoa), and Capitella sp. (Lophotrochozoa, Annelida). An E-value threshold was determined empirically for each protein family by using the E-value associated with the closest paralogue retrieved from Blast of the fly protein against the fly genome (see Additional file 2: E-values used for each family, with names and accession numbers of the closest fly paralogues).
Figure 1. Summary of the occurrence of post-synaptic proteins in the investigated genomes. Yellow fields indicate presence of an orthologue as determined from the corresponding gene tree topology. Letters correspond to the three additional confidence criteria for orthology assignments defined in the Methods section: b, ML bootstrap value >= 70% and/or Bayesian posterior probability >= 0.95; c, congruence between at least two partitions of the same protein or between different domains; d, conservation of domain architecture. Note that Homo and Drosophila have been used as reference taxa for defining the orthology groups.
Additional file 1. Detailed overview of the results from the phylogenetic analyses. Grey lines: presence or absence of orthologue(s) for each protein group in each species. White lines: number of orthologues (if any) identified for each alignment or alignment partition.
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Additional file 2. Table indicating the threshold E-values used for Blast searches for each family of investigated genes, and the closest paralogue retrieved from the fly genome.
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Alignments and phylogenetic analyses
Sequences retrieved from Blast searches were automatically integrated into the corresponding alignments and aligned using CLUSTALW, implemented in our in-house automated Blast and alignment pipeline. Alignments were slightly corrected manually for elimination of major mistakes. Ambiguous regions were identified by visual inspection and removed manually and positions containing more than 80% of missing data were deleted. When several non-overlapping partial sequences were present in the same alignment, the alignment was split into several partitions to avoid artificial grouping in the trees of non-overlapping sequences (observed in preliminary Maximum Likelihood analyses). These partitions, labelled "A, B, C, ..." (see Additional file 1), were analysed separately. The alignments are given in Additional file 3.
Additional file 3. Protein sequence alignments used for phylogenetic analyses. The alignments are in Phylip format.
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Maximum-Likelihood (ML) analyses were performed using the PhyML program [39], with the WAG model of amino-acid substitution and a BioNJ tree as the input tree. A gamma distribution with four discrete categories was used in these ML analyses. The gamma shape parameter and the proportion of invariant sites were optimised during the searches. Branch support was tested with bootstrapping (100 replicates). Bayesian analyses were done with Mr Bayes under the WAG model (prset=invgamma). For each data set, two searches were run in parallel for 500,000 generations, representing 20 times the number of generations discarded as burn in (first 25,000 trees, determined by stabilisation of the likelihoods). We estimated convergence by checking the "average standard deviation in partition frequency values across independent analyses", using a threshold value of 0.05. Since for most alignments the number of phylogenetically informative positions is low with respect to the number of sequences, trees contain a high number of unstable nodes and therefore it was not possible to constrain searches for convergence with a lower threshold value. In some cases, this threshold of 0.05 was not reached after 500,000 generations (and concomitantly stationarity of the likelihoods was not reached after 25,000 generations). Then, burn-in was expanded and search was prolonged until the index had dropped below 0.05. Posterior probabilities were estimated by constructing a majority-rule consensus of trees sampled every 100 generations. Trees were rooted as in ref [2].
After preliminary (not shown) phylogenetic analyses, all (protein/species) absences were further tested by additional Blast searches on the full (when possible, assembled) nucleotide genome sequences of the corresponding species, with a very relaxed E-value (10) to ensure that these genes have not been missed due to limitations of the initial Blast searches (i.e. the use of predicted transcript data sets, or too strict E-values). In several cases, these genomic searches detected protein orthologues that had been missed during initial searches on predicted transcripts (e.g. IP3R in Hydra magnipapillata; PMCA in Capitella sp.). In these cases, protein alignments were updated using the procedure described above, prior to final phylogenetic analyses (trees shown in Additional file 4).
Additional file 4. Maximum Likelihood phylogenetic trees of the post-synaptic proteins and of the PDZ, SAM and SH3 domains (arranged in alphabetic order). Maximum likelihood boostraps (ML bs) and Bayesian posterior probabilities (Bay pp) are indicated only for the main orthology groups. Abbreviations for taxon names: Aqu = Amphimedon queenslandica, Ath = Arabidopsis thaliana, Bde = Batrachochytridium dendrobatidis, Cca = Capitellasp., Ddi = Dictyostelium discoideum, Dme = Drosophila melanogaster, Hma = Hydra magnipapillata, Hsa = Homo sapiens, Mbr = Monosiga brevicolis, Ncr = Neurospora crassa, Nve = Nematostella vectensis, Osa = Oryza sativa, Ror = Rhizopus oryzae, Sce = Saccharomyces cerevisiae, Tad = Trichoplax adhaerens, Uma = Ustilago maydis. Except for plant, yeast, slime mold, sponge, fly and human sequences, genes are named by using a neutral code consisting in the taxon abbreviation followed by a letter (different for each protein family) and a number. Thanks to this label, a given gene can be identified across different partitions of the same alignment or different domains of the same protein (e.g. for Shank in the PDZ, SAM and SH3 trees).
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Orthology assignments
An orthology group is here defined as the clade containing all sequences that are more closely related to the fly and/or human sequence(s) of reference than to any paralogue of ancient origin (i.e. resulting from duplication before the last common ancestor of bilaterians). Our orthology assessments therefore rely primarily on gene tree topologies, and the yellow fields in Fig. 1 indicate the presence of an orthologue, based on the corresponding gene tree. Three additional criteria were taken into account to estimate confidence for these orthology assignments: (i) support values (ML bootstrap support >= 70% and/or Bayesian posterior probability >= 0.95) ("b" in Fig. 1), (ii) congruence between separate analyses of partitions of the same protein alignment (labelled A, B, ...: see above) or between different domains of the same protein ("c" in Fig. 1), (iii) comparison of domain architectures reconstructed using Smart http://smart.embl-heidelberg.de/ webcite and Pfam http://pfam.sanger.ac.uk/search webcite. Domain architecture was considered as conserved ("d" in Fig. 1) if the main functional domain(s) existing in the human and fly proteins were present and showed similar arrangement. These domain reconstructions are shown in Additional file 5. They were systematically done for the Monosiga and Trichoplax putative orthologues of human PSD proteins. In addition, we compared the five C-ter residues of PDZ-binding proteins in search of PDZ ligand conservation (Additional file 6), but the extent of sequence variability observed among PDZ ligands across taxa and proteins is so high that in practice it is generally impossible to conclude either for or against presence of a PDZ ligand. In addition, intron positions and frames were determined for the Shank proteins using NCBI sequence viewer (human and fly proteins) or JGI browse tool (proteins from Capitella, Nematostella and Monosiga) and the alignment procedure described above.
Additional file 5. Domain architectures of selected members of the post-synaptic gene families (in alphabetic order). Abbreviations used in displays for taxon names are as in Additional file 2.
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Additional file 6. The C-ter five residues of the PDZ-binding post-synaptic proteins, for metazoan species and Monosiga.
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Reconstruction of protein acquisitions and losses onto the taxon phylogeny
Presence/absence of the PSD proteins across our species sampling was formalised into a taxon/character matrix. These characters were mapped onto the taxon phylogeny under parsimony and a Dollo model of gene gain/loss (each protein has been acquired only once - no convergence - and proteins can be lost but not re-acquired - no reversion). We used the taxon phylogeny of [34], as it is the most comprehensive phylogenomic study of basal metazoan relationships to date. This means in particular that our interpretation of the data relies upon a hypothesis of eumetazoan monophyly (here represented by Cnidaria + Bilateria). However, basal metazoan relationships remain controversial (see alternative views in [28,29,31]), and the reconstruction may differ under alternative topologies. Since the relationships between sponges, Trichoplax and eumetazoans (here represented by cnidarians and bilaterians) were unsupported in ref [34], the reconstruction was done under the three possible tree topologies.
Results and Discussion
Reconstructing evolutionary acquisitions and losses of the major PSD components
Starting from the PSD protein and domain alignments of ref [2] we have extended the taxonomic coverage by adding genes retrieved from the complete genomes of several additional fungal species, the choanoflagellate Monosiga brevicolis, the placozoan Trichoplax adhaerens, the cnidarian Hydra magnipapillata and the annelid Capitella sp. Protein/domain orthology was determined based on gene trees (Additional file 4) and comparison of domain structures (Additional file 5). Results are summarised in Fig. 1, with a more detailed overview in Additional file 1. The most parsimonious reconstruction of protein acquisitions and losses is shown in Fig. 2 (see less parsimonious reconstructions obtained for alternative placements of Trichoplax, in Additional file 7). The inferred order of emergence of the genes is indicated by a colour code in Fig. 2 and Fig. 3. In Fig. 2, gene gains/losses are labelled in bold when orthology is supported by at least two criteria of confidence (in addition to gene tree topology; see Fig. 1 and Methods); otherwise they are labelled in normal font.
Figure 2. Gains (coloured dashes) and losses (ellipses) of post-synaptic proteins reconstructed onto the taxon phylogeny under the parsimony criterion. Placozoans are here placed as the sister-group to other metazoans. The total number of gains + losses is 45 (see less parsimonious reconstructions obtained for alternative placements of placozoans in Additional file 6). Events are labelled in bold when the corresponding gene orthology is supported by at least two confidence criteria in addition to gene tree topology (see explanations in Methods); otherwise they are labelled in normal font.
Figure 3. Graphic representation of the mammalian post-synaptic density, with the origination period of the genes indicated by the same colours as in Fig. 2. Domain structures are represented only for scaffold proteins. For most PDZ-binding proteins, the PDZ ligand has been labelled in grey, because the extent of sequence variability observed among PDZ ligands across taxa and proteins is so high that it is generally impossible to conclude either for or against the presence of a PDZ ligand (except for CRIPT and δ-cat; see Additional file 4). A darker red colour has been used for the Shank SAM domain, as it might not have been present in the Shank protein of the common choanoflagellate-metazoan ancestor (see text). Note that the general configuration of the post-synaptic scaffold has been completely re-drawn with respect to [2] (their Fig. 1), to account for data available in the recent literature (e.g. [17,18]).
Additional file 7. Gains (coloured dashes) and losses (ellipses) of post-synaptic proteins reconstructed with placozoans as the sister-group to sponges, and with placozoans as the sister-group to eumetazoans. The number of gains + losses is indicated for each tree.
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PSD components originated in two main steps
Protein acquisitions are concentrated in the two branches preceding and following the divergence of choanoflagellates in the opisthokont phylogeny (Fig. 2). A significant protein set originated in an exclusive ancestor of choanoflagellates and metazoans. This includes the scaffold proteins Shank, Homer and DLG, and the other PSD components CamKII, Citron, Cortactin, Ephrin R, IP3R, PKCαβγ and Shaker K+ channels. Another set, including the scaffold proteins CASK, GKAP and S-SCAM (= Magi), and the other PSD components α and δ-catenin, classical Cadherins, NOS, LIN-7, PICK1 and SPAR, is synapomorphic for the Metazoa. Very few proteins are of more ancient origin: PMCA (present in all investigated eukaryotes), β-catenin (present in plants and fungi) and CRIPT (present in plants).
A significant implication is that several core components of the mammalian PSD (red colour in Fig. 3) originated not only before the acquisition of synapses and nerve cells (absent in sponges and placozoan), but even before the acquisition of multicellularity, given that the choanoflagellate-metazoan ancestor was unequivocally unicellular [1,8]. Notably, the scaffold proteins Shank, Homer and DLG/PSD95, known in mammalians to structure the post-synaptic density by linking a huge variety of target proteins (e.g. membrane receptors, components of signal transduction pathways, cytoskeleton-interacting proteins, etc.), originated before the divergence of the choanoflagellates. In our analyses, the orthology of Homer and DLG between choanoflagellates and metazoans is well supported by the corresponding gene trees, the conservation of their domain structure, high bootstrap support (for Homer) and the congruence between the evolutionary histories of the domains (for DLG) (see Fig. 1).
Shank emerged before the divergence of choanoflagellates
The Shank protein is considered as the "master-scaffolding" molecule of the PSD [17,18]. Our PDZ tree identifies a putative choanoflagellate orthologue of metazoan Shank PDZs (PDZ tree in Additional file 4). This orthology assignment is not supported by statistical indices, but several additional lines of evidence argue in favour of orthology. The Monosiga protein that contains the putative Shank PDZ shows the combination of protein-interacting domains typical of vertebrate Shanks (N-ter ankyrin repeats, a SH3 domain, a PDZ domain, and a SAM C-ter domain), a domain structure that is unique among vertebrate proteins. Reciprocal Blast searches support orthology between the ankyrin repeats present in this Monosiga protein and those of metazoan Shanks (not shown - note that phylogenetic analyses of ankyrin repeats are not feasible). In addition, the putative Monosiga Shank protein shares with metazoan Shanks the presence of a conserved N-terminal 126-amino-acids region (upstream of the Ankyrin repeats) with no similarity to any other protein sequence (not shown result of Blast search against NCBI referenced proteins) (see alignment of this N-terminal region in Additional file 8). Within this region, there are two introns with position conserved down to the phase and present in metazoan Shanks as well as in the Monosiga protein (red boxes in the alignment, Additional file 8). Finally, we analysed SH3 domains and found that the SH3 domain of the Monosiga putative Shank falls in a clade that contains all metazoan Shank SH3s (and two non-Shank SH3s, from Drosophila and Hydra) (see SH3 tree in Additional file 4).
Additional file 8. Comparison of intron positions in the Shank proteins of metazoans and Monosiga. Overview of intron distribution along the whole protein sequences (A) and detailed view of intron position on the alignment of the N-terminal region upstream of the Ankyrin repeats (B).
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These observations indicate that Monosiga has a bona fide orthologue of Shank. Although there is no experimental evidence that in the choanoflagellate cell Shank acts like its mammalian orthologues, the conserved domain combination suggests comparable scaffold properties. In addition, some of the proteins known in mammalians to interact directly with Shank (including Homer, the IP3 receptor, and Cortactin) are also present in Monosiga.
The history of Shank has been slightly more complex in the details. Our phylogenetic analyses suggest that the SAM C-ter domain of Monosiga Shank might not be orthologous to the SAM domains present in some (but not all) metazoan Shanks (see SAM tree in Additional file 4). A SAM domain might thus have been added independently in choanoflagellate and metazoan Shanks. This convergence event could have occurred by chance, or it may have been favoured by similar functional constraints acting on the Shank protein. A second peculiarity of choanoflagellate Shank is that its SH3 domain is located between PDZ and SAM, whereas in metazoans it is placed between the Ank repeats and the PDZ domain, indicating that a domain inversion took place in one of these two lineages. We compared intron positions along the Shank proteins of metazoans and Monosiga without finding evidence for or against this hypothesis of domain inversion (Additional file 8).
Evolution of the PSD protein repertoire within the Metazoa
Compared to the gene complement present in eumetazoan genomes, different gene subsets happen to be absent in the two nerve-less species Amphimedon queenslandica (sponge) and Trichoplax adhaerens (placozoan). This makes the reconstruction of acquisition/losses in this region of the tree highly sensitive to the phylogenetic placement of placozoans with respect to sponges and eumetazoans. Trichoplax lacks orthologues of Shank (present in Monosiga, sponges and eumetazoans), GRIP, ErbB R, mGluR and SynGAP (present in sponges and at least some eumetazoans). If Trichoplax is the sister-group to other metazoans (Fig. 2), then GRIP, ErbB R, mGluR, and SynGAP were absent in the metazoan ancestor and are synapomorphic for sponges + eumetazoans (brown colour in Fig. 2 and 3). The same taxon phylogeny implies secondary loss of α-catenin and CASK (present in Trichoplax and in eumetazoans) in the sponge lineage. Alternative taxon phylogenies (e.g. Trichoplax sister-group to eumetazoans as in [34]) imply secondary losses of GRIP, ErbB R, mGluR and SynGAP in placozoans (Additional file 7). Whatever the position of placozoans, the parsimony optimisation suggests secondary loss of Shank in Trichoplax adhaerens.
The apparent loss of Shank in placozoans is particularly puzzling as Shank is the main protein responsible for the higher-order structure of the PSD in mammals [18]. However, alternative scenarios cannot be ruled out at this stage, e.g., Shank could have been overlooked due to incomplete coverage of the T. adhaerens genome (currently 8×), or the gene might have diverged in the placozoan lineage to the extent that it is no longer recognisable, or it might have been lost in Trichoplax adhaerens but not in other placozoan species. If confirmed by future studies, loss of Shank in placozoans would imply disruption or at least considerable alteration of the macromolecular architecture of the PSD-like protein complex. In addition, the Trichoplax situation is somewhat difficult to understand when it is realised that Homer and Cortactin, known as direct Shank partners in mammals and present in the Monosiga genome, are still present in Trichoplax despite apparent absence of Shank. As an alternative possibility, Shank could have been replaced by another protein. Functional work is certainly needed in Trichoplax to determine the molecular interactions and cellular functions of these proteins. Whether these genes are expressed in the fibre cells, proposed by some [40] to represent a placozan neural-like cell type, will be important to assess. However, placozoans are not likely to teach us much about the ancestral properties and functions of PSD proteins prior to the emergence of a nervous system, because the absence of Shank (if confirmed) makes their PSD-like complex totally idiosyncratic.
Whereas the emergence of nerve cells and synapses in a eumetazoan ancestor was seemingly not accompanied by a significant expansion of the PSD complex in terms of protein number, the few proteins acquired in this branch are significant with respect to neural function. They include the ionotropic glutamate receptors NMDAR (involved in synaptic transmission) and the Neuroligins (involved in synaptic adhesion and signalling in synaptogenesis, through interaction with their ligands the Neurexins [41,42]). Within eumetazoans, we inferred a few protein losses: ErbB receptor and GRIP (a scaffold protein) in cnidarians, α-catenin and Neuroligin in Hydra, SPAR in protostomians, Erbin in Drosophila and ErbB receptor in the annelid Capitella. Curiously enough, there is no concomitant loss of proteins known to interact with each other (e.g. losses of Erbin are not associated with losses of the ErbB receptor), suggesting that proteome and interactome evolved to a large extent independently, or that protein interactions in non-model organisms differ substantially from what happens in the mammalian synapse.
What were the ancestral functions of the PSD protein complex?
Comparative genomics contributes to our understanding of hierarchy in biological systems, by sorting ancient components from more recent additions, a basis for making predictions about ancestral interactions and functions. In bilaterian animals, the PSD is tightly linked functionally to intercellular signalling at the synapse. However, the proto-PSD of the choanoflagellate/metazoan ancestor (red + violet colours in Fig. 3) was probably not involved in intercellular signalling, since all post-synaptic membrane receptors linked to the mammalian PSD (the different kinds of glutamate receptors, Neuroligin, ErbB receptor, receptor domain of the Ephrin receptor) are of more recent origin. In addition, the global structure of the ancestral proto-PSD, which lacks several important scaffold proteins (GRIP, S-SCAM, CASK), was certainly much simpler than that of the mammalian PSD.
Shank and its conserved direct or indirect partners appear to form the ancestral core of the post-synaptic protein complex (Fig. 3). The associated functions are actin cytoskeleton regulation (Homer links actin to Shank; the actin-regulating protein Cortactin binds to Shank) and Ca2+ signalling (Homer links the IP3 receptor to Shank; CaMKII and PKCαβγ, also involved in Ca2+ signalling, are present in the choanoflagellate genome). It is thereby proposed here that the main function of the ancestral Shank interactome was to provide a link between intracellular Ca2+ signalling and actin cytoskeleton regulation.
The DLG scaffold protein might have ancestrally formed the centre of an additional interactome, possibly independent from the Shank module (GKAP, the physical link between DLG and Shank in mammals, is absent in Monosiga). We identified three DLG partners with clear orthologues in the choanoflagellate genome: CRIPT (a microtubule-interacting protein [43]), the K+ channel Shaker, and the Ca2+ plasma membrane channel PMCA. The choanoflagellate orthologue of Citron (a Rho target) is not likely to interact with DLG as in mammalians because only the most N-ter part of the protein shows homology with mammalian Citron (Fig. 3). These observations suggest ancestral functions of the DLG interactome in concentrating cation channels on particular plasma membrane areas, and in providing a link between these channels and the regulation of microtubule polymerisation.
From a morphological point of view, the choanoflagellate + metazoan clade is characterised by a unique synapomorphy: acquisition of an apical cell complex with a central flagellum surrounded by a collar of microvilli. This is suggested by the close resemblance between apical cell complexes of choanoflagellates and of sponge choanocytes [44]. According to some [45], this flagellum-collar structure would be homologous with the apical complex of eumetazoan sensory cells. Because choanoflagellate collar microvilli are contractile and labile structures (suggesting highly dynamic regulation), and because their cytoskeleton is principally made of F-actin, it is tempting to speculate about a possible functional link between the emergence of the proto-PSD and the acquisition of the flagellum-collar apical complex. This hypothesis can be tested by determining the subcellular localisation and function of the key proteins Shank, Homer and DLG in Monosiga. Another stimulating path for future investigation on the ancestral functions of core PSD components will involve the characterisation of physical interactions between the various choanoflagellate PSD-like proteins.
The backbone of the post-synaptic density, notably featuring the scaffold proteins Shank, Homer and DLG, was acquired in a common ancestor of choanoflagellates and metazoans, in a unicellular context. It was probably not involved in cell-cell communication, but rather in linking transmembrane cation currents, calcium signalling, and cytoskeleton regulation. Future investigations on complete genome sequences of unicellular opisthokonts that are more closely related to the choanoflagellate + metazoan clade than to Fungi (ichthyosporids and Capsaspora) [1] might happen to push even earlier the period of origination of these proteins.
After the addition of several novel components (among which GKAP, the link between DLG and Shank, and the CASK/S-SCAM/classical cadherin interactome), metazoan ancestors became endowed with a nearly complete set of PSD proteins (violet + red + blue colours in Fig. 3). Proteins that are specifically involved in synaptic transmission (i.e. AMPAR, NMDAR and mGluR) or adhesion (neuroligin) in eumetazoans were nevertheless absent from this ancestral animal repertoire (except possibly for mGluRs, present in the metazoan ancestor if Trichoplax is not the basal-most lineage, see Additional file 7). This suggests that PSD proteins ancestrally acted in a non-synaptic context, in consistence with the probable monophyly of Eumetazoa [34] and its corollary, that the last common ancestor of all metazoan animals was nerve-less. In this ancestor, the PSD-like complex was probably a sub-membrane platform for bringing together molecules involved in cation currents, Ca2+ and other signalling pathways, and cytoskeleton regulation, possibly (but not necessarily) in relation to cell sensitivity to external stimuli. The complex later became integrated into the evolving synapse, in a eumetazan ancestor, through the addition of novel proteins and functionalities.
Authors' contributions
AA and MM conceived and designed the analyses. AA performed the data collection and the analyses. MM drew the figures. AA and MM wrote the paper. Both authors read and approved the final manuscript.
We thank Sylvie Retaux and Hervé Le Guyader for insightful advice. We are grateful to Philippe Lopez for help with the phylogenetic analyses. This work was supported by a grant from the GIS "Institut de la Génomique Marine" - ANR "programme blanc" NT_NV_52 Genocnidaire.
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Foot pain causes
Alternative Names
Causes of foot pain
What causes foot pain?
Many things can cause foot pain. Wearing shoes that don't fit right is one of the most common reasons people (especially women) get foot problems such as calluses and bunions. With age, feet often grow wider. Also, being overweight can increase your chances of having foot problems or injuries. Go shopping for shoes in the afternoon when your feet are at their largest due to swelling. Heels on shoes shouldn't be higher than 1 inch. Shoes with a wide toe box can also lead to less problems to your feet.
See also:
Abu-Laban RB, Ho K. Ankle and foot. In: Marx JA, Hockberger RS, Walls RM, et al, eds. Rosen's Emergency Medicine: Concepts and Clinical Practice. 7th ed. Philadelphia, Pa: Mosby Elsevier; 2009:chap 55.
Borom AH, Clanton TO. Foot and ankle: Sports shoes and orthoses. In: DeLee JC, Drez D Jr, Miller MD, eds. DeLee and Drez's Orthopaedic Sports Medicine. 3rd ed. Philadelphia, Pa: Saunders Elsevier; 2009:chap 25, section B.
Koenig MD. Foot and ankle: Ligament Injuries of the Foot and Ankle in Adult Athletes. In: DeLee JC, Drez D Jr, Miller MD, eds. DeLee and Drez's Orthopaedic Sports Medicine. 3rd ed. Philadelphia, Pa: Saunders Elsevier; 2009:chap 25, section C.
Last reviewed 6/4/2011 by Linda J. Vorvick, MD, Medical Director, MEDEX Northwest Division of Physician Assistant Studies, University of Washington, School of Medicine; and C. Benjamin Ma, MD, Assistant Professor, Chief, Sports Medicine and Shoulder Service, UCSF Dept of Orthopaedic Surgery. Also reviewed by David Zieve, MD, MHA, Medical Director, A.D.A.M., Inc.
- The information provided herein should not be used during any medical emergency or for the diagnosis or treatment of any medical condition.
- A licensed medical professional should be consulted for diagnosis and treatment of any and all medical conditions.
- Call 911 for all medical emergencies.
- Links to other sites are provided for information only -- they do not constitute endorsements of those other sites.
©1997 - A.D.A.M., Inc.Any duplication or distribution of the information contained herein is strictly prohibited.

subset: commoncrawl


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St. Lucie 3B Richard Lucas hit a game-winning home run last night, but it's been a tough year (.212/.288/.350) for the 21-year old.
The eight home runs are nice, but he has just four doubles, an almost shockingly small number for a guy who's played 62 games. He's also struck out 74 times. That's a lot. I liked his strength and approach when I saw him last year, but clearly, the jump to the FSL has been hard for him.
In some ways, Lucas is the poster-child for the old regime and their haphazard player movement at the lower levels necessitated by pushing so many teenagers to the SAL. After a fine start to his pro career in the GCL in 2007, and seven games with Kingsport, the Mets pushed him to Savannah as a 19-year old in 2008. It was a lost season: he hit .185/.252/.311 and tore up his knee as a Gnat when he should have spent the whole year as a K-Met. Returned to Kingsport last year at age 20, he raked, going .357/.471/.622 in 26 games before struggling in the final two weeks at Brooklyn. With Jefry Marte ticketed for a return engagement in Savannah, the Mets had to push Lucas to St. Lucie to get him at bats in a move one team official described as "almost unfair" to him.
I wonder where he would be now had he gone 07 – GCL, 08 – Kingsport (uninjured), 09 – Savannah (while Marte played in a short-season league) before reaching St. Lucie this year.


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Will Self and Benjamin Zephaniah are among the authors appearing at the Hillingdon Literary Festival
Have you always wanted to catch a glimpse of famous constellations or distant galaxies? Now you can, with public events at Ruislip Lido!
The inmates received very realistic treatment and were fingerprinted, photographed and had DNA samples taken, before being placed in the cell
London Air Ambulance attended after reports of a man coming off his motorcycle on Eastcote Road
What's On writer Emily Chudy
enjoys a French meal in the newly refurbished Café Rouge in Ruislip
Looking for a fun night out locally this weekend? Join The Octavia Blues Band as they come to The Tropic at Ruislip!
The blaze destroyed the whole of the first floor, roof and half of the ground floor of the derelict building
Planning to let your hair down this weekend? Check out Hughie Dixon's latest round-up of live music in and around Hillingdon borough
President Andrzej Duda laid a wreath at the newly opened Remembrance garden at the Polish war memorial
Police officers tried to chase 22-year-old Luke Esprit but had to stop when he sped down Field End Road the wrong way
Frank Arens began performing from an early age, attending a Ruislip dance school from the age of three
London Ambulance Service took the driver of one of the vehicles to hospital after firefighters cut them free
Bank Holiday Monday opening times for Aldi, Asda, Tesco, Sainsbury's, Morrisons, Lidl and Waitrose
Ian Partridge discovered he had holes in his heart which he said affects one in six of the adult population
People all of ages flocked to Ruislip to experience medieval life with knights, jousting and re-enactments
Find out what's going in your area this weekend for fun ideas of things to get up to!
Find out what's going in your area this week for fun ideas of things to get up to!
Do you enjoy dancing and fancy meeting new people? A Dance Parties Plus event might be for you
Police said Gary Vines repeatedly offended over a period of two years including stealing from cars, shoplifting, being drunk in the street, and assaulting a shop keeper
2015 marks the year Paul Speck Jnr would have turned 21, so the family wanted to thank the charity-run hospital that helped them through his brain stem tumour treatment
ROS StageWorks Summer School has been awarded the National Operatic and Dramatic Association London YOUNGSTARS trophy for its production of Beauty and The Beast Jr.
See video of fly-though animation of the proposed route HS2 would take through west London, from the sky
This week is National Parks Week! Head to one of these lovely green spots near you and enjoy nature
Cast a look back at these amazing TV shows set in our very own west London – and tell us what others should make the list...
Want to know if your MP supports the fox hunting ban or wants to see it repealed? Use our gadget to find out their views on the controversial subject
Ruislip petitioners believed the HS2 tunnel would be 30m below their houses but documents reveal it would be HALF that
More than a 100 firefighters tackled the blaze in Wadsworth Road, Perivale, which sent smoke billowing across London. Around 30 people at the site which includes factory units escaped before the fire crews arrived
Residents want something to be done to stop speeding on their street, but say traffic islands will "hinder, not help" them
The dance school's 87-year-old founder said: "It's quite an honour to have been able to do it for all those years"
Ruislip is part of the London Borough of Hillingdon. Major landmarks in the area include Ruislip Lido, a former reservoir, now an area of public parkland with its own miniature railway and Manor Farm, a settlement dating from the 9th century which is now designates as a local heritage site.
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All efforts to stop the deadly impact of global warming will ultimately fail. Yes, fail. Our leaders are destroying us from within. Blame GOP, conservatives, Big Oil, capitalist billionaires, Koch Bros. Also the president. Harper's quotes John Podesta, Obama's senior adviser, warning: "History will judge Obama's climate-change efforts as sadly insufficient."
America's leaders are failing us, will never act, till it's too late to plan for an economic catastrophe. We have lost our soul.
There are four big reasons our leaders will not act in time: First. Overwhelming opposition by the GOP, conservatives and capitalists ... Second. Our blind failure to see our planet cannot feed the 10 billion people predicted by 2050. ... Third. Our addiction to "more is never enough" consumption ... Fourth. The irrational, unscientific and ultimately self-destructive belief of capitalism in perpetual economic growth, on a planet of finite resources.
We know most will continue believing in all four, trapped in denial. Eventually reality will expose our fantasies, but not soon enough. Global warming will continue causing irreparable damage, destroying our world, pushing us past the point of no return and driving us into a "chaotic era" where few humans will survive.
Here are the details, the four reasons why we're now an addicted nation that can't stop self-destructing as we've reached a critical mass that's sabotaging all efforts to stop global warming:
1. Relentless opposition of conservative capitalists to climate science
Conservative opposition is everywhere, emanating from the economic fears of the energy industry, Big Oil, coal, gas, other polluters. They fear regulation and taxation of carbon emissions will have a major negative impact on their corporate earnings, shareholder dividends and will increase production costs, limit access to resources.
They spend hundreds of millions lobbying politicians and it works. California Gov. Jerry Brown says there's "virtually no Republican" in Washington that accepts climate science and most GOP governors "openly deny climate science," despite widespread scientific evidence.
Worldwide, conservative leaders like Australian Prime Minister Tony Abbott celebrated the repeal of his country's carbon-emission tax, despite the increasing damage to his country's environment and over the unanimous objections from Anglican Church leaders opposing repeal .
In fact, even as the new U.S. EPA regulations were announced, Abbott was touring the world to create a "conservative alliance among 'like-minded' countries, aiming to dismantle global moves to introduce carbon pricing and undermine a push by America to push the case for action through forums such as the G20."
Conservatives are getting even stronger, neutralizing efforts by power-players like Michael Bloomberg, Hank Paulson, billionaire Tom Steyner and their "RiskyBusiness.org" project intended to educate businesses and politicians on the real costs and "economic risks from the impacts of a changing climate." Intense opposition from conservative capitalists like the Koch Bros. is also there to blunt Risky Business and similar climate-science advocates.
And what about Obama's new EPA regulations? The impact will be watered down, much as conservatives did with big bank regulations after the 2008 disaster. Plus, global warming regs will kick in too slowly, get watered down by Big Oil lobbyists. Moreover, in spite of all the scientific evidence, sadly, surveys reveal widespread public apathy about future damage to the planet. It's not a big motivator for most. What is? Cheap gas for their cars is a higher priority.
2. Out-of-control population is the biggest driver of global warming
The United Nations predicts global population will exceed 10 billion by 2050, adding three billion more human beings in one short generation, on a planet of rapidly dwindling resources. Plus science is already warning us we cannot feed everybody. In his new book, "The Countdown," Alan Weisman, author of an earlier bestseller, "World Without Us," asks the taboo question no politician can ask: "How many people can our planet hold?"
Weisman's answers are deadly: What if science confirms that a "sustainable population is even less than the 7 billion we already number ... how do we design an economy for a shrinking, and then a stable one, meaning, an economy that can prosper without depending on constant growth?" Weisman also offers a solution: "Social objections aside, if the entire world adopted a one-child policy" then by 2100, we'd "be back to 1.6 billion, our population in 1900." But politicians will be silent ... till it's too late.


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|Four Star Running Back Commits To Heels
|Written by Staff Reports
|Wednesday, 11 July 2012 16:00
The state's top-ranked running back has made a verbal commitment to play for North Carolina, according to his school's athletic director. T.J. Logan from Northern Guilford High in Greensboro made his pledge to UNC on Tuesday afternoon over offers from Auburn, California, Clemson, N.C. State, Ohio State, South Carolina (runner-up), Tennessee,
Virginia, Wake Forest and West Virginia.
|Share This Article:


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Lil Wayne feat. Drake & Jadakiss – "Its Good" (Shots Fired At Jay-Z)
Just incase you have been hiding under a rock or something, above is the track that has the internet going crazy. With the release of "Carter IV" just a few days away, here is the newest offering featuring Drake and Jadakiss from the album. As you can see in the title, shots are fired at Mr. Shawn Carter. This could get interesting. Well that is if Hov decides to respond.
"Talking about that Baby money, I got your baby Money / Kidnap your bitch, keep that how much you love your lady money"


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Canadian mobile phone maker Research In Motion is gearing up for the launch of its new BlackBerry 10 platform on January 30, but it seems that some lucky people will be able to learn more on the platform earlier than that.
The handset vendor will hold a pre-launch event together with wireless carrier T-Mobile on January 24, where it will unveil some more info on what its new OS has to offer, gizmofusion notes.
Apparently, the meeting will take place in New York in a secret location, yet no specific info on who will attend has been provided as of now.
T-Mobile is one of the wireless carriers in the United States set to add the new BlackBery 10 devices to its offering as soon as RIM makes them available but, as of yet, it hasn't offered specific info on when they would be launched.


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200-Hour Yogaworks Teacher Training
spiritual, strong, fun
It is an extension of my dance background. I like the meditative and healing properties of yoga. Like dance yoga has been around for centuries.
I love seeing students find their wholeness and center.
Walking on the beach, Swimming in the Ocean, Breastlink organization, Mind/Body/Soul


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(Tyra's makeover episodes not included.)
Hair Apparent
Clinique Superdefense Triple Action Moisturizer SPF 25 "makes skin soft and silky" and acts as a "shield against sun and pollution" to help prevent brown spots and fine lines.
You "tried all the others" and fell in love with "skin-tone fixer" Olay Definity Correcting Protective Lotion with SPF 15
Lash Out


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3. Fitness Physiology L+L
The purpose of this course is to acquire a basic understanding of how the human body functions to maintain a state of wellness. We will explore the short-term responses to exercise and discuss how the body responds to long-term exercise (training) programs. At the end of the course, students should be able to examine the design of exercise physiology experiments, as well as understand and interpret reports of health and exercise news in the popular press. Laboratory 15 hours. THIS COURSE SATISFIES THE LABORATORY SCIENCE UNIVERSITY CORE REQUIREMENT.
4. Light & Life L+L
This course focuses on the importance of plants and photosynthesis for the future of life on earth. Issues addressed include: food production, plants as renewable energy sources, "greenhouse effect" and using plants to clean up toxic wastes. In the laboratory students design experiments to examine the effects of environmental changes on photosynthesis. Laboratory 15 hours. THIS COURSE SATISFIES THE LABORATORY SCIENCE UNIVERSITY CORE REQUIREMENT.
5. Endangered Ecosystems L+L
This course provides an overview of earth's ecosystems and the major factors contributing to the loss of biodiversity. We will cover three major themes: 1) general ecological principles, especially focused on the structure and function of ecosystems, 2) factors contributing to the endangerment of ecosystems, and 3) the conservation of ecosystems and biodiversity. Although we will cover a number of global environmental problems, several lectures will highlight current environmental and conservation issues here in California and within the SF Bay Area. Laboratory 15 hours. THIS COURSE SATISFIES THE LABORATORY SCIENCE UNIVERSITY CORE REQUIREMENT.
6. Oceans L+L
Examination of major ocean ecosystems and their inhabitants, with special attention paid to issues of governmental policy and management, fisheries, and global population effects. Laboratory activities will emphasize local and regional marine habitats and human impacts on regional ecosystems. Laboratory 15 hours. THIS COURSE SATISFIES THE LABORATORY SCIENCE UNIVERSITY CORE REQUIREMENT.
15. The Human Embryo L+L
This course will explore two major themes. The first is to acquire a basic understanding of the biology of human reproduction and development. The second is to investigate how our basic knowledge of human reproduction is being used by medical science to assist in reproductive processes and correct developmental errors. Case-based discussions will focus on topics that include genetic screening, stem cell research, in vitro fertilization, and environmental toxins and their effects on embryo development. Laboratory experiments will be linked to the case studies to illustrate the techniques and issues raised by these topics. Laboratory 15 hours. THIS COURSE SATISFIES THE LABORATORY SCIENCE UNIVERSITY CORE REQUIREMENT.
18. Exploring Biotechnology L+L
Exploration of comtemporary biotechnology and the underlying science; how DNA, genes and cells work. Laboratory experiments focus on DNA in a variety of contexts. Laboratory 15 hours. THIS COURSE SATISFIES THE LABORATORY SCIENCE UNIVERSITY CORE REQUIREMENT.
19. Biology for Teachers L+L
Specifically designed for candidates for Multiple Subject Teaching Credentials. Provides an overview of the life sciences, focusing on physiology and cell biology, ecology, genetics, and evolution. In addition, laboratory experiences will introduce students to the scientific method, experimental design, data collection and analysis, and communication. Laboratory 15 hours.
21. Introduction to Physiology
Introduction to general principles underlying homeostasis, and the relationship of anatomical form to biological function. The course will introduce students to the organization and function of cells, cellular metabolism, energy, nutrition, regulation, communication, gas exchange, circulation, and osmoregulation. Prerequisite: Completion of or concurrent enrollment in CHEM 11. THIS COURSE SATISFIES THE LABORATORY UNIVERSITY CORE REQUIREMENT.
22. Introduction to Evolution & Ecology
Introduction to key concepts in evolution and ecology, including population genetics, natural selection and adaptation, phylogenetics and biodiversity, demography, and interactions among organisims and their environments. Prerequisite: Biology 21.
23. Investigations in Evolution & Ecology L+L
Introduction to experimental and statistical approaches used in modern ecological and evolutionary studies, with an emphasis on experimental design, data analysis, interpretation and presentation. Builds on concepts presented in BIO 22. Fieldwork and laboratory exercises (30 hours) will take advantage of the diversity of local terrestrial and marine ecosystems. Prerequisites: Biology 22.
24. Introduction to Cellular & Molecular Biology
An introduction to the cell and molecular fundamentals necessary for life. Topics include macromolecular structure, enzyme function, membrane structure and physiology, metabolism and bioenergetics, the cell cycle, and classical and molecular genetics. Prerequisites: Biology 21 and completion of or concurrent enrollment in Chemistry 31.
25. Investigations in Cellular & Molecular Biology L+L
An introduction to experimental methods for studying the cellular and molecular basis of life. Builds on the concepts covered in Biology 24. Topics include enzyme function and kinetics, cell reproduction, Mendelian genetics and molecular biology. The topics are explored through laboratory work, with emphasis placed on the analysis, interpretation, and presentation of experimental data. Laboratory 30 hours. Prerequisites: Biology 24 and completion of or concurrent enrollment in Chemistry 32.
99. Special Topics
Investigation of a specific area or topic in the biological sciences. Open to majors and non-majors. Pre-requisite: approval of department chair.
101. Biology Research Seminar
A forum for the exploration of active research themes in the life sciences. Invited scientists from a range of universities and institutes present their current research and engage in discussion about this research with seminar participants. This course is intended to give students direct interactions with research academics in a range of fields, to make them aware of career opportunities and to provide them with contacts in those fields. Students may take the course more than once for credit but BIOL 101 does not count as one of the seven upper-division biology courses required for the major. Graded P/NP only. Prerequisite: Successful completion of BIOL 23, 24, or 25. (2 units).
104. Human Anatomy L+L
An exploration of the structure, organization, and functional relationships of human anatomical systems. (Laboratory dissections use alternative vertebrates.) Laboratory 30 hours. Prerequisite Biology 25.
106. Health Consequences of a Western Lifestyle
This course explores the impact of living in a developed country on human health. Topics such as diabetes, obesity, heart disease, hypertension, and cancer will be discussed at the molecular, cellular, physiological, and population levels. Prerequisite: BIOL 25. Also listed as PHSC 124. (5 units)
110. Genetics L&L
Basic principles governing inheritance and gene expression in viruses, prokaryotes, and eukaryotes. Emphasis on molecular aspects. Laboratory 30 hours. Prerequisite: Biology 25.
113. Microbiology L&L
An introduction to the biology of microorganisms, with emphasis on the molecular and cellular biology of bacteria, the diversity of microbial life, and the roles of microorganisms in human health and disease. Laboratory 30 hours. Prerequisite: Biology 25.
114. Immunology L&L
Principles, mechanisms, and techniques of humoral and cellular aspects of the immune response. Immediate and delayed hypersensitivity, tissue transplantation, tumor immunology, and immunodeficient states in humans. Laboratory 30 hours. Prerequisite: Biology 25.
115. Human Reproduction & Development
Detailed study of the development and function of the male and female reproductive systems, gametogenesis, fertilization and implantation as well as an anatomical description of the heart, circulatory, nervous and skeletal systems during embryogenesis. Where appropriate the molecular mechanisms controlling the determination of these developing systems will be examined. Prerequisite: Biology 24.
116. Medical Microbiology L&L
This upper division course focuses on the interactions of pathogenic microbes (bacteria, viruses, fungi, prions, etc.) with their hosts. We will examine the various strategies employed by the infectious agents to subvert the immune system and the various strategies used by the immune system to combat the microbial invasion. We will also examine the co-evolution of hosts and their pathogens and the natural history of diseases. The laboratory component will expose students to clinical methodologies and scientific approaches to diagnose and differentiate pathogenic microorganisms. Prerequisite: BIOL 25.
117. Epidemiology L&L
This course provides an introduction to epidemiology, including assessment of health and disease in populations, epidemiological data analysis, disease transmission, and public health interventions. The course also exposes students to the epidemiology of diseases and conditions of current public health and clinical importance in the United States and internationally. Laboratory 30 hours. The laboratory (computer lab) will provide students with hands-on experience with epidemiologic methods, study design, and data analysis. Also listed as PHSC 100. Prerequisite: BIOL 24. (5 units)
119. Biology of Stress
This course explores the impact of stress on physiology, behavior, and health, using a multidisciplinary approach. Topics include defining and measuring stress, differences between acute and chronic stress exposure, effects of stress on physiological processes and on the brain, how stress affects gene expression and neurogenesis, and relationships between stress and disease. We will also discuss the social patterning of stress exposure and the effects of social policies and interventions. Prerequisite: BIOL 24. (5 units)
120. Animal Physiology L&L
Examination and comparison of mechanisms used by a variety of animals to survive and function in their environment. Laboratory 30 hours. Prerequisite: Biology 24.
122. Neurobiology L&L
An introduction to the nervous system, focusing on the anatomy, physiology, and pharmacology of neurons, synapses, and simple neural circuits. Laboratory 30 hours. Prerequisite: Biology 25.
124. Human Physiology L&L
Examining the physical and chemical basis of human life, this course focuses on the neural and endocrine control of physiologic processes to maintain homeostasis. Laboratory 30 hours. Prerequisite: Biology 25.
128. Plant Development L+L
Developmental processes of plants, with emphasis on current research and experimental approaches. Laboratory 30 hours. Prerequisite: Biology 25.
134. California Plant Diversity
The California flora includes more than 5,800 species of plants, of which about one-quarter are only found here. In this course, we will investigate the ecological and evolutionary underpinnings of this diversity through a combination of lecture and extensive field-based lab activities. Content includes identification of California's major plant families, how to identify species with a dichotomous key, and how molecular phylogenetic studies are contributing to our understanding of major ecological and evolutionary themes in the California flora.
145. Virology
Biology of viruses: their structure, evolutionary origins, classification, genetics, laboratory propagation and diagnostic methods, viral pathogenesis, response of host cells to viral infectiion and salient aspects of the epidemiology of viral diseases. The focus will be on viruses that infect eukaryotic cells, emphasizing important viral groups that infect humans. Prerequisite: Biology 25. Recommended: Chemistry 141.
151. Restoration Ecology L+L
The science and practice of restoring degraded ecosystems, with an emphasis on plant ecology. Through fieldwork in restoration experiments and examination of literature case studies, students will grapple with basic questions: How do we decide what to restore? How do we restor it? And how do we know if we're finished? Emphasis on reading and writing scientific papers, working with data, and critically judging the success of restoration projects in meeting goals of biodiversity and ecosystem function. Laboratory and field work 30 hours. Also listed as ENVS 151. Prerequisite: Biology 23.
153. Conservation Science
Conservation is a scientific enterprise and a social movement that seeks to protect nature, including Earth's animals, plants, and ecosystems. Conservation science applies principles from ecology, population genetics, economics, political science, and other natural and social sciences to manage and protect the natural world. Conservation is all too often seen as being at odds with human well-being and economic development. This course explores the scientific foundations of conservation while highlighting strategies to better connect conservation with the needs of a growing human population. We will examine whether conservation can protect nature, not from people, but for people. Also listed as ENVS 153. Prerequisite: BIOL 23. (5 units)
156. General Ecology L+L
Quantitative study of the interrelationships of organisms with their biotic and abiotic environments. Emphasis on population dynamics, interspecific relationships, community structure, and ecosystem processes. Laboratory and field work 30 hours, including one weekend field trip. Prerequisites: Biology 23 and Math 11.
158. Biology of Insects L&L
An introduction to basic and applied aspects of insect biology, with emphasis on evolution, morphology, physiology, and behavior of insects and related arthropods. Also includes a review of important agricultural, medical, forestry, and veterinary pests. Laboratory 30 hours. Prerequisite: Biology 24.
160. Biostatistics L+L
A course in applied statistics for biologists planning to conduct manipulative experiments. Biologists gain training in experimental design and quantitive analyses in cases where specific experimental hypotheses are to be tested. Theory and concepts are covered in lectures and readings. Laboratory sessions provide practical experience in computing statistical procedures by hand and with statistical software. Examples used in lectures and lab assignments are derived from medical research, physiology, genetics, evolution, ecology, and environmental risk assessment. Laboratory 30 hours. Prerequisite: Biology 23. Also listed as Environmental Studies 110.
165. Animal Behavior L&L
Examination of the behavior of animals in nature using an organizational scheme that recognizes proximate, or immediate, causes of behavior and evolutionary bases for behavior. Topics include physiological correlates of behavior, perception of natural stimuli (light, sound, chemicals); and behavioral ecology of foraging, mating systems, parent-offspring relationships, and social behavior. Laboratory 30 hours. Prerequisite: Biology 24.
171. Ethical Issues in Biotechnology and Genetics
An interdiscipinary consideration of contemporary biotechnology and the ethical implications inherent in the development and use of such technology. Topics include human cloning, stem cell research, human genome project, genetic testing gene therapy, genetically modified organisms, personalized medicine, clinical trials, and public policy. Biology 171 satifies a biotechnology minorrequirement but NOT the Ethics requirement. When taken concurrently with Biology 189 it satisfies an upper-division biology major requirement. It also fulfills the third Religious Studies requirement Prerequisite: Biology 24 or permission of instructor. Biology 25 strongly recommended.
173. Evolution L+L
Examination of advanced concepts of modern evolutionary biology. Topics include the evolutionary forces of microevolution, the evolution of sex, adaptation, speciation, human evolution, molecular evolution and macroevolutionary phenomena deciphered from phylogenetic trees. Laboratory experiments, field study, and computational activities 30 hours. Prerequisite: BIOL 24. Recommended: BIOL 110.
174. Cell Biology L+L
Study of the function of cellular organelles and the signaling pathways that control cell reproduction. Topics include a detailed discussion of the structure of cell membranes, nuclear and chromosome structure, DNA replication, the microtubule and microfiliment cytoskeleton, mitosis, mechanisms of cell motility, cell cycle regulation, and apoptosis. Laboratory experiments focus on cell cycle regulation and cell differentiation. Laboratory 30 hours. Prerequisite: Biology 25.
175. Molecular Biology L+L
An introduction to the molecular events involved in the replication and decoding of the genetic material. Lectures focus on basic molecular biology concepts and recombinant DNA technology. Laboratory 30 hours. Prerequisite: Biology 25.
176. Biotechnology Lab I: Recombinant DNA L&L
Explores techniques for the anaylsis of DNA, and the construction and manipulation of recombinant DNA molecules. Laboratory meets twice each week. Lectures discuss the scientific basis for the lab methods, and their application in biomedical research and the biotechnology industry. Laboratory 60 hours. Prerequisites: Biology 25 and at least one upper-division biology laboratory course. (Does not include field courses). Biology 175 recommended.
177. Biotechnology Lab II: Gene Expression and Protein Purification L&L
Explores principles and techniques for the expression and purification of recombinant proteins. Laboratory meets twice each week and will use techniques such as column chromatography, mammalian tissue culture, and various gene expression systems. Lectures discuss the theory behind the methods used in lab as well as their application in basic and applied research. Laboratory 60 hours. Prerequisites: Biology 25 and at least one upper division Biology laboratory course (Does not include field courses). Biology 175 recommended.
178. Bioinformatics L&L
Bioinformatics tools are important for storing, searching, and analyzing macromolecular sequences and structures. This course in applied bioinformatics provides an in-depth survey of modern bioinformatics tools. Students will become proficient at searching GenBank, downloading and analyzing sequences, working with metadata and each student will write an original computer program to complete an independent research project. Software tools for functional and evolutionary analysis of nucleic acids and proteins will also be examined. Prerequisite: Biology 25. Biology 175 recommended.
179. Cancer Biology
Introduction to the molecular and cellular basis of cancer. Introduction to the pathology of cancer. How basic processes such as cell growth, cell cycle control, and cell death are affected by molecular changes in onco-genes and tumor-suppressor genes. Laboratory uses molecular and cytogenic tools important in cancer diagnosis.
180. Marine Physiological Ecology
Examination of the physiology of major groups of marine animals and of relationships between animals and their environment. Laboratory and field work (30 hours) focuses on the diverse marine ecosystems of California. Prerequisite: Biology 24.
189. Topics in Cell and Molecular Biology
Seminar dealing with contemporary research in cellular and molecular biology and biotechnology. Students are required to lead discussions and participate in the critical analysis of recently published research articles. Does NOT count as an upper-division course toward a major or minor in biology but allows Biology 171 to count as an upper-division biology course for the Biology major or minor when Biology 189 and 171 are taken during the same quarter. Biology 189 may be taken up to two times for credit Prerequisites: Completion of concurrent enrollment in Genetics, Cell Biology, Microbiology, or Molecular Biology. Students who have completed Biology 25 are welcome to attend and participate in the discussion of these topies but may not take the course for credit until they have completed one of the prerequisites.
190A and 190B. Contemporary Issues in Biology
Specialized treatment of some aspect of biology of current interest to the biologist as well as to society in general. Prerequisites will be specified according to topic.
191. Project Lab L+L
An innovative laboratory course in which undergraduate students will participate in a supervised research project directly related to ongoing research in the laboratory of the principal investigator. Each year, the course will focus on research activities appropriate for the particular stage of the research project. Course can only be taken once. Laboratory 60 hours. Prerequisites: Biology 25 (required) and at least one upper-division Biology Lab course (recommended).
195. Undergraduate Research
Experimental research project supervised by Biology Department faculty. Five hours of research per week is expected per unit. Maximum of 3 units per quarter. Can be repeated for credit, with a maximum of 5 units per academic year. Must be taken P/NP. Prerequisite: Consent of instructor.
198. Internship and Undergraduate Research Students wishing to take either 198A or 198B should have a GPA of 3.0 or better in biology and must present an outline of their projected research to the chair not later than the fifth week of the term preceding the start of the project. Prerequisite: Departmental and University permission.
198A. Internship
Research in off-campus programs under the direct guidance of cooperating research scientists and faculty advisers.
198B. Research
Supervised laboratory research culminating in a written report suitable for publication. Sustained for one year with credit given for one term.
199. Directed Reading and Research
Detailed investigation of a specific topic in biology under the close direction of a faculty member. Students wishing to take this course should have a GPA of 3.0 or better in biology and must present an outline of their projected research to the department chair not later than the fifth week of the term preceding the start of the project, which will continue for one term only. Prerequisite: Departmental and University permission.


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Assassin's Creed IV: Black Flag Review
Assassin's Creed IV: Black Flag Review: Experience The Thrilling Combat on both Sea and Land!
Finally, the new part of Assassin's Creed series has been released. New expansion named Assassin's Creed IV: Black Flag is offering us many breathtaking experiences and very intense and attractive combat. After getting the opportunity to try the game and experience many things which the game offers, I have decided to share my impressions about the game with you, so here is the Assassin's Creed IV: Black Flag Review.
Right from start, we take the role of Edward Kenwey and start the magnificent journey overseas. That's right, in this expansion, sailing has amazing role, it is not just a side thing like in previous part. Now, you own your own ship which will be your loyal companion together with your crew on your adventure.
There are plenty of things to do, besides following the main story and conquering numerous fortresses and various other places, freeing slaves and killing enemies in exciting combat. You are free to sail as you wish, you are a real pirate and you can loot any ship you come across.
By doing so, you will earn wanted reputation and you will be attacked by pirate hunters. Piracy and constant action on water will let you experience combat that you have not seen before. Using ship cannons, mortars, boarding on enemy ships and looting their treasures is something that you have to experience.
Even the ground missions are great. You can use your environment much more effective than in previous parts of Assassin's Creed. All those stealthy kills and assassinations that your enemies would never expect will make your blood boil.
Your character, Edward Kenwey, is master of every weapon, so you can freely pick up and loot weapons from both your enemies and killed allies and use them in combat. There are various throwing darts that will provide you with many possibilities which will assist you in combat. Are you in trouble, do you have low health and you have to escape? Use smoke dart and you will be able to take down your enemies and escape without trouble! Is there some sniper on tower that is carefully watching the spot through which you have to pass? No problem, hit him with sleeping dart and he will take a short nap while you sneak through that spot safely. Do you enjoy watching the enemies fight each other? Use berserk dart and create conflicts in enemy's camp.
There are so many ways to fight, lots of stuff were taken from earlier parts of game, like using enemy as a meat shield, but there are bunch of innovations as well.
Thrilling combat with all those exciting moves and possibilities are definitely amazing and they will make you enjoy every moment you spend playing Assassin's Creed IV: Black Flag. Piracy and sea sailing, upgrading your ship and fighting many enemies on both sea and land, even upgrading your own pirate hideout, purchasing supplies and looting enemy's ships together with numerous other actions will provide you with unique experience.
Even though there is so much action and everything, it's sad that you have not so many options to affect the story by yourself, so you mostly just have to follow it, but all in all, it is really exciting and worth playing game!
Country: Canada
Release Date: 29 October 2013 (USA)


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Phoenix Applicants
CHCI's R2L NextGen is an exciting opportunity available to students from select geographic areas where Ready to Lead (R2L) events have been held. This page can provide you more information for Phoenix area students interested in applying for the program.
I live in the Phoenix area. How do I know if I'm eligible?
Not sure what county you live in? Click the link to visit the county website, which list all the towns within it.
While attendance at previous Phoenix Ready to Lead events is not required in order to apply, attendance at Ready to Lead will be considered during the selection process. For more information about Ready to Lead, click here.
In addition to meeting one of the geographic requirements, students must:
- Be a current 9th, 10th, or 11th grade student
- Demonstrate interest in learning more about civic engagement
While we do require applicants share their grades and a copy of their transcript, there is no minimum GPA requirement to apply.
Interested in applying? Visit apply.chci.org and complete the application named "R2L NextGen - Phoenix Applicants." Applications must be completed by April 30, 2014 at 11:59 pm Eastern Standard Time. That's 9:59 pm Mountain Standard Time!
Do you have additional questions? Contact Evelyn Garcia Morales, Manager of Educational Enrichment Programs at email@example.com.


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Survey Shows Customers Do Not Enjoy Shopping Online
The fun of shopping online has worn off. Online megamall iStorez studied how people feel about shopping online. Only 1 in 10 Americans now shop just 'for the fun of it,' and the majority want improvements.
Here are some more findings:
- 78% of younger shoppers say they think online shopping has room for improvement.
- 55% find online shopping more convenient than shopping in stores
- 36% find greater financial rewards (look online to get bargains)
- 35% find more variety offline (even though there is much more variety online)
- The woman who has the most admired fashion sense in politics right now is former vice president candidate Sarah Palin. That came at a price though – she famously $150,000 on her wardrobe.
The study, conducted by Kelton Research, might mean that online shopping may slow down, just as people are spending less in stores. Anand Jagannathan, CEO of iStorez points out: "The truth is that the online shopping experience hasn't changed much in the last decade. If shoppers are telling us that online shopping offers less variety than real-world malls, then we need to listen hard and make some major changes." I suppose that means offer more selection?
What do people want? First, 55% say they shop online for the convenience of it. But they want retailers to streamline and simplify the process. To that I'd add that shopping for clothes online that you haven't tried can take a long time. It's a pain to return things that don't fit (unless they work with their offline store to make it easier).
iStorez carries 18 million products from 500 retailers. Customers can create a own personal mall based on their style and favorite stores.
About the Author: Janet Meiners always wanted to be a reporter but prefers the immediacy of blogging. Known as Newspapergrl, she has freelanced for newspapers, City Search, and business magazines. She reports on internet marketing and online trends. MarketingPilgrim posts


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Electronic Equipment, Instruments and Components
Company Overview of Atlassian, Inc.
Executive Profile
Erik K. Bardman
Chief Financial Officer, Atlassian, Inc.
|Total Calculated Compensation
|This person is connected to 1 Board Members in 1 different organizations across 5 different industries.
See Board Relationships
Mr. Erik K. Bardman has been Chief Financial Officer at Atlassian, Inc. since February 2015. Mr. Bardman has more than 20 years of global business experience in financial services and Internet commerce, focusing on pricing, mergers and acquisitions, strategic development, and financial planning and analysis. He served as Chief Financial Officer of Roku, Inc., since April 2013. Mr. Bardman served as the Chief Financial Officer and Senior Vice President of Finance at Logitech ...
Corporate Headquarters
375 Alabama StreetPhone: 415-701-1110
San Francisco, California 94110
United States
Fax: 415-449-6222
Board Members Memberships
Independent Director, Chairman of Audit Committee, Member of Nominating & Corporate Governance Committee and Member of Strategic Transactions Committee
Bachelor's Degree
Dickinson College
Other Affiliations
Annual CompensationThere is no Annual Compensation data available.
Stocks OptionsThere is no Stock Options data available.
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|There is no Competitor Compensation data available.
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Our data partners will research the update request and update the information on this page if necessary. Research and follow-up could take several weeks. If you have questions, you can contact them at firstname.lastname@example.com.


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48,XXYY syndrome
- 48,XXYY Klinefelter syndrome
- 48,XXYY variant of Klinefelter's syndrome
- XXYY syndrome
On this page
Newline Maker
What are signs and symptoms of 48,XXYY syndrome?
48,XXYY syndrome can affect other parts of the body as well. Males with 48,XXYY syndrome are often taller than other males their age. They tend to develop a tremor that typically starts as a young adult and worsens with age. Dental problems are frequently seen with this condition; they include delayed appearance of the primary (baby) or secondary (adult) teeth, thin tooth enamel, crowded and/or misaligned teeth, and multiple cavities. As affected males get older, they may develop a narrowing of the blood vessels in the legs, called peripheral vascular disease. Peripheral vascular disease can cause skin ulcers to form. Affected males are also at risk for developing a type of clot called a deep vein thrombosis (DVT) that occurs in the deep veins of the legs. Additionally, males with 48,XXYY syndrome may have flat feet (pes planus), elbow abnormalities, allergies, asthma, type 2 diabetes, seizures, and congenital heart defects.[1][2]
Most males with 48,XXYY syndrome have some degree of difficulty with speech and language development. Learning disabilities, especially reading problems, are very common in males with this disorder. Affected males seem to perform better at tasks focused on math, visual-spatial skills such as puzzles, and memorization of locations or directions. Some boys with 48,XXYY syndrome have delayed development of motor skills such as sitting, standing, and walking that can lead to poor coordination. Affected males have higher than average rates of behavioral disorders, such as attention deficit hyperactivity disorder (ADHD); mood disorders, including anxiety and bipolar disorder; and/or autism spectrum disorders, which affect communication and social interaction.[1]
- US National Library of Medicine. 48,XXYY syndrome. Genetics Home Reference. January 2010; http://ghr.nlm.nih.gov/condition/48xxyy-syndrome.
- Chen H.. Klinefelter syndrome. e-medicine Medscape. May 17, 2011; http://emedicine.medscape.com/article/945649-followup#a2650. Accessed 6/23/2011.
- 48,XXYY Klinefelter syndrome
- 48,XXYY variant of Klinefelter's syndrome
- XXYY syndrome


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In addition, RNA isolated from malignant glioma cells using the StrataPrep microprep kit has been used in differential display experiments. These experiments identified several differentially expressed genes in the invasive rim of the tumor relative to the noninvasive tumor core.4
The StrataPrep total RNA microprep kit is ideal for the isolation of RNA from small cell samples harvested by LCM. The method generates pure, intact RNA with high efficiency that can be effectively amplified by conventional and quantitative RT-PCR.
The authors thank Michael Berens, Neurology Research, Barrow Neurological Institute, Phoenix, AZ for providing LCM samples.
1. Emmert-Buck, M. R., et al. (1996) Science 274:
2. Dolter, K. and Braman J. (2000) Strategies 13: 56-58.
3. Rogers, B. and McKenzie, D. (1998) Strategies 11: 100-102.
4. Berens, M. E., et al. (2000) Proceedings of the American Association for Cancer Research 41:232, Abstract 1477.


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Book your room at Apartamentos Rurales Complejo Peñafiel
Apartment, 1 BedroomMax Occupancy: 3 guests (up to 2 children, 1 infant)
Apartment, 2 BedroomsMax Occupancy: 6 guests (up to 5 children, 1 infant)
Get an overview of this hotel3-star country house with restaurant and coffee shop/café
Situated in a rural location, this country house is 14.5 mi (23.3 km) from Puente Romano and within 32 mi (52 km) of Convento de San Benito and Coria Castle. Coria Cathedral is 25.5 mi (41 km) away.
Property Features
Along with a restaurant, this country house has a coffee shop/café and self parking. WiFi in public areas is free. Other amenities include a meeting room, free use of bicycles, and dry cleaning.
Room Amenities
This country house has 9 rooms.Information missing or incorrect? Tell us!
Hotel Amenities
Hotel Amenities
Dining options at the country house include a restaurant and a coffee shop/café. Public areas are equipped with complimentary high-speed wireless Internet access. This Zarza la Mayor country house also offers a terrace, tour/ticket assistance, and multilingual staff. Onsite parking is available (surcharge).
- Breakfast available (surcharge)
- Coffee shop or café
- Free newspapers in lobby
- Dry cleaning/laundry service
- Elevator/lift
- Free WiFi
- Laundry facilities
- Luggage storage
- Multilingual staff
- Number of buildings/towers - 1
- Number of floors - 3
- One meeting room
- Restaurant
- Self parking (surcharge)
- Terrace
- Total number of rooms - 9
- Tours/ticket assistance
Available in some public areas: Free WiFi
Self parking (surcharge)
Room Amenities
- Air conditioning
Where to Eat
Apartamentos Rurales Complejo Peñafiel has a restaurant on site.
Nearby Things to Do
The recreational activities listed below are available either on site or nearby; fees may apply.
- Free bicycles on site
- Bicycle rentals nearby
- Hiking/biking trails nearby
Hotel Policies
Check-in time starts at 2 PM
Minimum check-in age is 18
Check-in time ends at midnight
Check-out time is noon
Payment types
Children and extra beds
- Children are welcome.
- Rollaway/extra beds are available for EUR 15 per night.
Pets allowed
You need to know
Extra-person charges may apply and vary depending on hotel policy.
Government-issued photo identification and a credit card or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
Optional extras
- Rollaway bed fee: EUR 15 per night
We should mention
Cash transactions at this property cannot exceed EUR 2500, due to national regulations. For further details, please contact the property using information in the booking confirmation.