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During Bacillus subtilis sporulation, the transient engulfment defect of spoIIB strains is enhanced by spoVG null mutations and suppressed by spoVS null mutations. These mutations have opposite effects on expression of the motility regulon, as the spoVG mutation reduces and the spoVS mutation increases [[sigma].sup.D]-directed gene expression, cell separation, and autolysis. Elevating [[sigma].sup.D] activity by eliminating the anti-[sigma] factor FlgM also suppresses spoIIB spoVG, and both flgM and spoVS mutations cause continued expression of the [[sigma].sup.D] regulon during sporulation. We propose that peptidoglycan hydrolases induced during motility can substitute for sporulation-specific hydrolases during engulfment. We find that sporulating cells are heterogeneous in their expression of the motility regulon, which could result in phenotypic variation between individual sporulating cells.

Research has been conducted on the forespore engulfment by the mother cell in Bacillus subtilis. The role of SpoIIB in engulfment has been investigated.

A 45% bandwidth in-line air-filled waveguide-to-substrate integrated waveguide (SIW) direct transition is designed to greatly improve the performance of existing configurations. The transition, based on a four-section height-stepped waveguide, includes a single-step widening transformer that enables full-band operation without using any dielectric probe. The absence of the probe reduces significantly the insertion loss and makes this transition simple, bandwidth controllable and easily scalable to the millimeter-wave frequency range. A back-to-back transition is designed to cover the 32–50 GHz band showing a return loss of 15 dB and a mean insertion loss of 0.68 dB in the whole bandwidth. [ABSTRACT FROM PUBLISHER]

With an increase in consumer demand of video gaming entertainment, the game industry is exploring novel ways of game interaction such as providing direct interfaces between the game and the gamers' cognitive or affective responses. In this work, gamer's brain activity has been imaged using functional near infrared spectroscopy (fNIRS) whilst they watch video of a video game (League of Legends) they play. A video of the face of the participants is also recorded for each of a total of 15 trials where a trial is defined as watching a gameplay video. From the data collected, i.e., gamer's fNIRS data in combination with emotional state estimation from gamer's facial expressions, the expertise level of the gamers has been decoded per trial in a multi-modal framework comprising of unsupervised deep feature learning and classification by state-of-the-art models. The best tri-class classification accuracy is obtained using a cascade of random convolutional kernel transform (ROCKET) feature extraction method and deep classifier at 91.44%. This is the first work that aims at decoding expertise level of gamers using non-restrictive and portable technologies for brain imaging, and emotional state recognition derived from gamers' facial expressions. This work has profound implications for novel designs of future human interactions with video games and brain-controlled games. [ABSTRACT FROM AUTHOR]

Lactococcus lactis is a promising candidate for the development of mucosal vaccines. More than 20 years of experimental research supports this immunization approach. In addition, 3' 5'- cyclic di-adenosine monophosphate (c-di-AMP) is a bacterial second messenger that plays a key role in the regulation of diverse physiological functions (potassium and cellular wall homeostasis, among others). Moreover, recent studies showed that c-di-AMP has a strong mucosal adjuvant activity that promotes both humoral and cellular immune responses. In this study, we report the development of a novel mucosal vaccine prototype based on a genetically engineered L. lactis strain. First, we demonstrate that homologous expression of cdaA gen in L. lactis is able to increase c-di-AMP levels. Thus, we hypothesized that in vivo synthesis of the adjuvant can be combined with production of an antigen of interest in a separate form or jointly in the same strain. Therefore, a specifically designed fragment of the trans-sialidase (TScf) enzyme from the Trypanosoma cruzi parasite, the etiological agent of Chagas disease, was selected to evaluate as proof of concept the immune response triggered by our vaccine prototypes. Consequently, we found that oral administration of a L. lactis strain expressing antigenic TScf combined with another L. lactis strain producing the adjuvant c-di-AMP could elicit a TS-specific immune response. Also, an additional L. lactis strain containing a single plasmid with both cdaA and tscf genes under the Pcit and Pnis promoters, respectively, was also able to elicit a specific immune response. Thus, the current report is the first one to describe an engineered L. lactis strain that simultaneously synthesizes the adjuvant c-di-AMP as well as a heterologous antigen in order to develop a simple and economical system for the formulation of vaccine prototypes using a food grade lactic acid bacterium.

Structural studies have implicated Cys(9), Cys(104) and Cys(207) of simian virus 40 (SV40) Vp1 in disulfide bond formation. Recently, we have shown the three cysteines to be essential for disulfide linkage of Vp1 complexes in vitro. Here, the role of the three cysteines was explored during the course of SV40 infection. Single-, double- and triple-mutant Vp1 at Cys(9), Cys(104) and Cys(207) continued to localize to the nuclei of transfected CV-1 cells and to bind DNA, but showed a range of abilities to form plaques. Only mutants containing the Cys(9)-->Ser change showed defects in plaque formation. Single mutants at Cys(9) formed small plaques; mutants at Cys(9). Cys(104), Cys(9). Cys(207) and Cys(9). Cys(104). Cys(207) formed no plaques. All three isolated revertants contained back-mutations at the Vp1 Cys(9) codon. These results further confirm the involvement of the three Vp1 cysteines in protein-protein interactions during virus assembly. Cys(9) is critical for production of wild-type infectious virions, whereas Cys(104) and Cys(207) play secondary roles.

Previous studies have implicated disulfide bonds between Vp1 molecules in the stabilization of the simian virus 40 (SV40) capsid. To identify the cysteine residues involved in intermolecular disulfide interactions, systematic oligo-directed mutagenesis of cysteine codons to serine codons was initiated. Wild-type and mutant Vp1 proteins were produced in rabbit reticulocyte lysates and were allowed to interact post-translationally. Disulfide-linked Vp1 complexes were assessed via non-reducing SDS-PAGE and via sucrose-gradient sedimentation. Wild-type Vp1 forms 7S pentamers followed by 12S disulfide-linked multi-pentameric complexes in cell-free lysates. Mutagenesis of all seven cysteine codons abolished Vp1 12S complexes, but did not affect pentamer formation. A quadruple Vp1 mutant at Cys49, Cys87, Cys254 and Cys267 continued to form 12S complexes, whereas the major products of the Cys9, Cys104 and Cys207 triple mutant Vp1 were 7S pentamers. Single and double mutant Vp1 proteins at the three cysteines affected continued to form 12S complexes, but to a lesser extent. Thus, inter-pentamer disulfide bonds at Cys9, Cys104 and Cys207 are essential and sufficient for stabilization of Vp1 complexes in cell-free lysates. These results are in agreement with previous structural studies of SV40 that implicated the same three residues in disulfide linkage in the capsid. Possible parameters for the involvement of the three cysteines are discussed.