SummaryType III secretion system (T3SS) tip complexes serve as adaptors that bridge the T3SS needle and the pore-forming translocation apparatus. In this report we demonstrate that Bsp22, the most abundantly secreted substrate of the Bordetella T3SS, selfpolymerizes to form the Bordetella bronchiseptica tip complex. Bsp22 is required for both T3SS-mediated cytotoxicity against eukaryotic cells and haemoglobin release from erythrocytes. Bacterial two-hybrid analysis and protein pull-down assays demonstrated the ability of Bsp22 to associate with itself and to bind BopD, a component of the Bordetella translocation pore. Immunoblot and cross-linking analysis of secreted proteins or purified Bsp22 showed extensive multimerization which was shown by transmission electron microscopy to lead to the formation of variable length flexible filaments. Immunoelectron microscopy revealed Bsp22 filaments on the surface of bacterial cells. Given its required role in secretion and cell-surface exposure, we tested the protective effects of antibodies against Bsp22 in vitro and in vivo. Polyclonal antisera against Bsp22 fully protected epithelial cells from T3SS-dependent killing and immunization with Bsp22 protected mice against Bordetella infection. Of the approximately 30 genes which encode the Bordetella T3SS apparatus, bsp22 is the only one without characterized orthologues in other well-characterized T3SS loci. A maximum likelihood phylogenetic analysis indicated that Bsp22 defines a new subfamily of T3SS tip complex proteins. Given its immunogenic and immunoprotective properties and high degree of conservation among Bordetella species, Bsp22 and its homologues may prove useful for diagnostics and next-generation subunit vaccines.

SummaryHere we review the Bordetella virulence secretome with an emphasis on factors that translocate into target cells. Recent advances in understanding the functions of adenylate cyclase toxin, a type 1 secretion system (T1SS) substrate, and pertussis toxin, a type IV secretion system (T4SS) substrate, are briefly described and a compilation of additional secretion systems and secreted factors is provided. Particular attention is devoted to the Bsc type III secretion system (T3SS) and controversies surrounding it. Efforts to identify effector proteins, characterize in vitro and in vivo phenotypes, and the potential role of type III secretion during human infections are discussed.

An investigation on the detailed kinetics of proton transfer between a set of di- and monofluoro- and chloro- (2,3-, 2,5-, 2,6-, 3,5-, 2-, and 3-) benzoic acids (HA) and Crystal Violet carbinol base in chlorobenzene favors a mechanism in terms of fast equilibrium between HA and D to form a H-bonded complex, D...HA, followed by rate-limiting proton transfer along the H-bond to form the colored ion pair DH+A- under the combined influence of monomer HA catalyst, nonreactive cyclic dimer (HA)2 inhibitor, and hyperacidic homoconjugated complex H(HA2) catalyst through a transition state with nearly 60% charge separation.

The Bhatia-Thornton (BT) correlation functions namely the number-number, concentration-concentration and number-concentration correlation functions in liquid binary melts are important parameters for understanding the complexities in binary liquids. In this paper, the microscopic BT correlation functions of liquid Al-Cu alloys are investigated using square well (SW) potential under mean spherical model approximation. These correlation functions are linearly related to the Ashcroft-Langreth type partial structure factors. Thermodynamically important microscopic function i.e. concentrationconcentration correlation function in the long-wavelength limit, S CC (0) is analytically calculated, which is well-known in BT formalism and is often used to characterized the mixing behavior in binary melts. The Warren-Cowley chemical short range order parameters, α as function of Cu concentration is also computed in the liquid Al-Cu alloys through structural studies in the long wavelength limit, which gives valuable information regarding ordering in melts. The compound forming behavior in Al-Cu alloys is evidenced by the results obtained for the value of S CC (0) and α . Further, transport properties in binary mixture have been determined by using BT-correlation functions and its long PAPER: Classical statistical mechanics, equilibrium and non-equilibrium

The results of measurements of substituent induced chemical shifts of carboxyl carbons (deltaCO) of dichloro- and difluorobenzoic acids, including the monosubstituted ones with substituents at meta- and/or ortho- positions, in chloroform-d and strengths of these acids (log K) in chlorobenzene show an anomalous reverse trend between deltaCO and log K, while the electron density at carboxyl carbons should influence similarly both deltaCO and log K. A detailed chemometric analysis of comparison of disubstituent effects between deltaCO and log K on the basis of Fujita-Nishioka's multiparameter approach and assumption of additivity of substituent effects shows a dominance of the localized pi-polarization mechanism relative to simple electrostatic effects upon deltaCO. Further, steric factors play a significant role in determining deltaCO whereas with respect to log K they were insignificant. The overall anomaly has been rationalized keeping in mind that, while log K is a gross measure of energy differences between the ionized and unionized forms of the acids, deltaCO is a very sensitive probe for determining changes in electron density at the carboxyl carbon of the unionized acid.