Biosynthesis of B-band lipopolysaccharide (LPS) in Pseudomonas aeruginosa follows the Wzy-dependent pathway, requiring the integral inner membrane proteins Wzx (O-antigen [O-Ag] flippase), Wzy (O-Ag polymerase), and WaaL (O-Ag ligase). For an important first step in deciphering the mechanisms of LPS assembly, we set out to map the membrane topology of these proteins. Random and targeted 3′ wzx, wzy, and waaL truncations were fused to a phoA-lacZα dual reporter capable of displaying both alkaline phosphatase and β-galactosidase activity. The results from truncation fusion expression and the corresponding differential enzyme activity ratios allowed for the assignment of specific regions of the proteins to cytoplasmic, transmembrane (TM), or periplasmic loci. Protein orientation in the inner membrane was confirmed via C-terminal fusion to green fluorescent protein. Our data revealed unique TM domain properties in these proteins, particularly for Wzx, indicating the potential for a charged pore. Novel periplasmic and cytoplasmic loop domains were also uncovered, with the latter in Wzy and WaaL revealing tracts consistent with potential Walker A/B motifs.

This paper reviews studies from the past 30 years that use operations research methods to tackle containership routing and scheduling problems at the strategic, tactical, and operational planning levels. These problems are first classified and summarized, with a focus on model formulations, assumptions, and algorithm design. The paper then gives an overview of studies on containership fleet size and mix, alliance strategy, and network design (at the strategic level); frequency determination, fleet deployment, speed optimization, and schedule design (at the tactical level); and container booking and routing and ship rescheduling (at the operational level). The paper further elaborates on the needs of the liner container shipping industry and notes the gap between existing academic studies and industrial practices. Research on containership routing and scheduling lags behind practice, especially in the face of the fast growth of the container shipping industry and the advancement of operations research and computer technology. The purpose of this paper is to stimulate more practically relevant research in this emerging area.

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The Singapore Strait is considered as the bottleneck and chokepoint of the shipping routes connecting the Indian and the Pacific Ocean. Therefore, the ship collision risk assessment is of significant importance for ships passing through the narrow, shallow, and busy waterway. In this paper, three ship collision risk indices are initially proposed to quantitatively assess the ship collision risks in the Strait: index of speed dispersion, degree of acceleration and deceleration, and number of fuzzy ship domain overlaps. These three risk indices for the Singapore Strait are estimated by using the real-time ship locations and sailing speeds provide by Lloyd's MIU automatic identification system (AIS). Based on estimation of these three risk indices, it can be concluded that Legs 4W, 5W, 11E, and 12E are the most risky legs in the Strait. Therefore, the ship collision risk reduction solutions should be prioritized being implemented in these four legs. This study also finds that around 25% of the vessels sail with a speed in excess of the speed limit, which results in higher potentials of ship collision. Analysis indicates that the safety level would be significantly improved if all the vessels follow the passage guidelines.

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