Intercellullar junctions formed by cadherins, including desmosomes and adherens junctions, comprise two dimensional arrays of ‘‘trans’’ dimers formed between monomers emanating from opposing cell surfaces. Lateral ‘‘cis’’ interfaces between cadherins from the same cell surface have been proposed to play a role in cadherin clustering. Although the molecular details of cis interactions remain uncertain, they must define an anisotropic arrangement where binding is favorable only in certain orientations. Here we report Monte Carlo simulations performed on a 2D lattice constructed to account for the anisotropy in cadherin cis interactions. A crucial finding is that the ‘‘phase transition’’ between freely diffusing cadherin monomers and dimers and a condensed ordered 2D junction formed by dimers alone is a cooperative process involving both trans and cis interactions. Moreover, cis interactions, despite being too weak to be measured in solution, are critical to the formation of an ordered junction structure. We discuss these results in light of available experimental information on cadherin binding free energies that are transformed from their bulk solution values to interaction energies on a 2D lattice.

In order to recognize the problems associated with the transport mechanism of containment during the ventilation process of a laboratory fume hood, a transparent, full scale chemical fume hood is constructed for experimental studies. Distributions of mean velocity and velocity fluctuation in the sash plane are measured using a thermal anemometer. Flow patterns and tracer-gas concentration leakages are respectively diagnosed via the laser-assisted flow visualization method and the EN 14175-3 test protocol. The magnitudes of measured velocity fluctuations exhibit a sharp peak along the perimeter of the sash opening. The results of flow visualization verify that the elevated turbulence fluctuations are induced by the boundary-layer separation when the flow passes over the edges of sash perimeter. The tracer gas experiment shows that the regions where high degree containment leakages detected are located along the perimeter of hood aperture. Eleven commercial hoods which are claimed with fine aerodynamic design are further tested for confirmation of these observations. The results show similar correlations. Conclusions thus are made that large-scale vortex structures occurring around the perimeters of hood aperture due to the boundary-layer separation could induce strong turbulence, and therefore enhance dispersion of the hood containment.

BACKGROUND: Rahnella is a widely distributed genus belonging to the Enterobacteriaceae and frequently present on vegetables. Although Rahnella has interesting agro-economical and industrial properties and several strains possess antibiotic resistances and toxin genes which might spread within microbial communities, little is known about plasmids of this genus. Thus, we isolated a number of Rahnella strains and investigated their complements of small plasmids. RESULTS: In total 53 strains were investigated and 11 plasmids observed. Seven belonged to the ColE1 family; one was ColE2-like and three shared homology to rolling circle plasmids. One of them belonged to the pC194/pUB110 family and two showed similarity to poorly characterised plasmid groups. The G+C content of two rolling circle plasmids deviated considerably from that of Rahnella, indicating that their usual hosts might belong to other genera. Most ColE1-like plasmids formed a subgroup within the ColE1 family that seems to be fairly specific for Rahnella. Intriguingly, the multimer resolution sites of all ColE1-like plasmids had the same orientation with respect to the origin of replication. This arrangement might be necessary to prevent inappropriate synthesis of a small regulatory RNA that regulates cell division. Although the ColE1-like plasmids did not possess any mobilisation system, they shared large parts with high sequence identity in coding and non-coding regions. In addition, highly homologous regions of plasmids isolated from Rahnella and the chromosomes of Erwinia tasmaniensis and Photorhabdus luminescens could be identified. CONCLUSIONS: For the genus Rahnella we observed plasmid-containing isolates at a frequency of 19%, which is in the average range for Enterobacteriaceae. These plasmids belonged to different groups with members of the ColE1-family most frequently found. Regions of striking sequence homology of plasmids and bacterial chromosomes highlight the importance of plasmids for lateral gene transfer (including chromosomal sequences) to distinct genera.

Cross-linking proteins can mediate the emergence of rigid bundles from a dense branched network of actin filaments. To enable their binding, the filaments must first bend towards each other. We derive an explicit criterion for the onset of bundling, in terms of the initial length of filaments L, their spacing b, and cross-linker concentration f, reflecting the balance between bending and binding energies. Our model system contains actin, the branching complex Arp2/3 and the bundling protein fascin. In the first distinct stage, during which only actin and Arp2/3 are active, an entangled aster-like mesh of actin filaments is formed. Tens of seconds later, when filaments at the aster periphery are long and barely branched, a sharp transition takes place into a star-like structure, marking the onset of bundling. Now fascin and actin govern bundle growth; Arp2/3 plays no role. Using kinetic Monte Carlo simulations we calculate the temporal evolution of b and L, and predict the onset of bundling as a function of f. Our predictions are in good qualitative agreement with several new experiments that are reported herein and demonstrate how f controls the aster-star transition and bundle length. We also present two models for aster growth corresponding to different experimental realizations. The first treats filament and bundle association as an irreversible sequence of elongation-association steps. The second, applicable for low f, treats bundling as a reversible self-assembly process, where the optimal bundle size is dictated by the balance between surface and bending energies. Finally, we discuss the relevance of our conclusions for the lamellipodium to filopodia transition in living cells, noting that bundles are more likely nucleated by ‘‘tip complex’’ cross-linkers (e.g. mDia2 or Ena/VASP), whereas fascin is mainly involved in bundle maintenance.

Many cell-cell adhesive events are mediated by the dimerization of cadherin proteins presented on apposing cell surfaces. Cadherin-mediated processes play a central role in the sorting of cells into separate tissues in vivo, but in vitro assays aimed at mimicking this behavior have yielded inconclusive results. In some cases, cells that express different cadherins exhibit homotypic cell sorting, forming separate cell aggregates, whereas in other cases, intermixed aggregates are formed. A third pattern is observed for mixtures of cells expressing either N- or E-cadherin, which form distinct homotypic aggregates that adhere to one another through a heterotypic interface. The molecular basis of cadherin-mediated cell patterning phenomena is poorly understood, in part because the relationship between cellular adhesive specificity and intermolecular binding free energies has not been established. To clarify this issue, we have measured the dimerization affinities of N-cadherin and E-cadherin. These proteins are similar in sequence and structure, yet are able to mediate homotypic cell patterning behavior in a variety of tissues. N-cadherin is found to form homodimers with higher affinity than does E-cadherin and, unexpectedly, the N/E-cadherin heterophilic binding affinity is intermediate in strength between the 2 homophilic affinities. We can account for observed cell aggregation behaviors by using a theoretical framework that establishes a connection between molecular affinities and cell-cell adhesive specificity. Our results illustrate how graded differences between different homophilic and heterophilic cadherin dimerizaton affinities can result in homotypic cell patterning and, more generally, show how proteins that are closely related can, nevertheless, be responsible for highly specific cellular adhesive behavior.

INTRODUCTION: Catatonia is a common presentation to psychiatric services in developing countries. Medical causes of catatonia are common and often missed. Identifying causes for catatonia is important not only to guide proper management but to determine prognostic outcomes CASE PRESENTATION: We report a case of a 20-year-old male who presented with catatonia. Subsequent investigations revealed a chronic subdural haematoma. Implications of late presentation to orthodox services are discussed. CONCLUSION: Careful clinical observation, investigation and a high index of suspicion are necessary to effectively manage this condition.

During cellular migration, regulated actin assembly takes place at the cell leading edge, with continuous disassembly deeper in the cell interior. Actin polymerization at the plasma membrane results in the extension of cellular protrusions in the form of lamellipodia and filopodia. To understand how cells regulate the transformation of lamellipodia into filopodia, and to determine the major factors that control their transition, we studied actin self-assembly in the presence of Arp2/3 complex, WASp-VCA and fascin, the major proteins participating in the assembly of lamellipodia and filopodia. We show that in the early stages of actin polymerization fascin is passive while Arp2/3 mediates the formation of dense and highly branched aster-like networks of actin. Once filaments in the periphery of an aster get long enough, fascin becomes active, linking the filaments into bundles which emanate radially from the aster’s surface, resulting in the formation of star-like structures. We show that the number of bundles nucleated per star, as well as their thickness and length, is controlled by the initial concentration of Arp2/3 complex ([Arp2/3]). Specifically, we tested several values of [Arp2/3] and found that for given initial concentrations of actin and fascin, the number of bundles per star, as well as their length and thickness are larger when [Arp2/3] is lower. Our experimental findings can be interpreted and explained using a theoretical scheme which combines Kinetic Monte Carlo simulations for aster growth, with a simple mechanistic model for bundles’ formation and growth. According to this model, bundles emerge from the aster’s (sparsely branched) surface layer. Bundles begin to form when the bending energy associated with bringing two filaments into contact is compensated by the energetic gain resulting from their fascin linking energy. As time evolves the initially thin and short bundles elongate, thus reducing their bending energy and allowing them to further associate and create thicker bundles, until all actin monomers are consumed. This process is essentially irreversible on the time scale of actin polymerization. Two structural parameters, L, which is proportional to the length of filament tips at the aster periphery and b, the spacing between their origins, dictate the onset of bundling; both depending on [Arp2/3]. Cells may use a similar mechanism to regulate filopodia formation along the cell leading edge. Such a mechanism may allow cells to have control over the localization of filopodia by recruiting specific proteins that regulate filaments length (e. g., Dia2) to specific sites along lamellipodia.

The binding of the myristoylated alanine-rich C kinase substrate (MARCKS) to mixed, fluid, phospholipid membranes is modeled with a recently developed Monte Carlo simulation scheme. The central domain of MARCKS is both basic (zeta = +13) and hydrophobic (five Phe residues), and is flanked with two long chains, one ending with the myristoylated N-terminus. This natively unfolded protein is modeled as a flexible chain of ‘‘beads’’ representing the amino acid residues. The membranes contain neutral (zeta = 0), monovalent (zeta = -1), and tetravalent (zeta = -4) lipids, all of which are laterally mobile. MARCKS-membrane interaction is modeled by Debye-Huckel electrostatic potentials and semiempirical hydrophobic energies. In agreement with experiment, we find that membrane binding is mediated by electrostatic attraction of the basic domain to acidic lipids and membrane penetration of its hydrophobic moieties. The binding is opposed by configurational entropy losses and electrostatic membrane repulsion of the two long chains, and by lipid demixing upon adsorption. The simulations provide a physical model for how membrane-adsorbed MARCKS attracts several PIP2 lipids (zeta = -4) to its vicinity, and how phosphorylation of the central domain (zeta = +13 to zeta = +7) triggers an ‘‘electrostatic switch’’, which weakens both the membrane interaction and PIP(2) sequestration. This scheme captures the essence of ‘‘discreteness of charge’’ at membrane surfaces and can examine the formation of membrane-mediated multicomponent macromolecular complexes that function in many cellular processes.