Myocardial segment-specific model generation for simulating the electrical action of the heart

Background  

Computer models of the electrical and mechanical actions of the heart, solved on geometrically realistic domains, are becoming an increasingly useful scientific tool. Construction of these models requires detailed measurement of the microstructural features which impact on the function of the heart. Currently a few generic cardiac models are in use for a wide range of simulation problems, and contributions to publicly accessible databases of cardiac structures, on which models can be solved, remain rare. This paper presents to-date the largest database of porcine left ventricular segment microstructural architecture, for use in both electrical and mechanical simulation.     

Crystal Structure of the cGMP-dependent Protein Kinase II Leucine Zipper and Rab11b Protein Complex Reveals Molecular Details of G-kinase-specific Interactions

cGMP-dependent protein kinase (PKG)-interacting proteins (GKIPs) mediate cellular targeting of PKG isoforms by interacting with their leucine zipper (LZ) domains. These interactions prevent aberrant signaling cross-talk between different PKG isotypes. To gain detailed insight into isotype-specific GKIP recognition by PKG, we analyzed the type II PKG leucine zipper domain and found that residues 40–83 dimerized and specifically interacted with Rab11b. Next, we determined a crystal structure of the PKG II LZ-Rab11b complex. The PKG II LZ domain presents a mostly nonpolar surface onto which Rab11b docks, through van der Waals interactions. Contact surfaces in Rab11b are found in switch I and II, interswitch, and the β1/N-terminal regions. This binding surface dramatically differs from that seen in the Rab11 family of interacting protein complex structures. Structural comparison with PKG Iα and Iβ LZs combined with mutagenic analysis reveals that GKIP recognition is mediated through surface charge interactions.     

Multi-species predator eradication within a predator-proof fence at Ka‘ena Point, Hawai‘i

Ka‘ena Point Natural Area Reserve on O‘ahu hosts one of the largest seabird colonies in the main Hawaiian Islands and supports three species of endangered plants. In order to stop chronic predation by invasive alien mammals on native species, a peninsula-style predator-proof fence was constructed around a 20-ha portion of the reserve in 2011. Multi-species predator removal efforts began upon fence completion; diphacinone poison in bait boxes spaced 25 m apart was used to remove black rats, house mice, and small Indian mongooses. House mice also were removed with multiple-catch live traps spaced 12.5 m apart. Feral cats were removed with padded leg-hold traps. Feral cats and mongooses were eradicated in 1 month, black rats were eradicated in 2.5 months, and house mice were eradicated in about 9 months. Since eradication, incursions of cats and mongoose have been rare (1/7.2 months), but incursion frequency has been higher for black rats (1/56 days) and house mice (1/36–47 days). Buffer predator control was conducted to limit predator access and prevent reinvasion around the fence ends along the shoreline. Even with the high initial fence cost and ongoing predator incursion management, this method is expected to become more cost effective than previous predator control efforts after 16 years. Record numbers of Wedge-tailed shearwaters and Laysan albatrosses have fledged from the reserve after predator eradication, and regeneration of native plants and invertebrates is being observed. With careful planning and persistence, predator fences can be a cost-effective method of protecting natural resources, and multiple species of predators can be eradicated with traps and first-generation anti-coagulents.     

A Differential Fluorescence-Based Genetic Screen Identifies Listeria monocytogenes Determinants Required for Intracellular Replication

Listeria monocytogenes is a Gram-positive, facultative intracellular pathogen capable of causing severe invasive disease with high mortality rates in humans. While previous studies have largely elucidated the bacterial and host cell mechanisms necessary for invasion, vacuolar escape, and subsequent cell-to-cell spread, the L. monocytogenes factors required for rapid replication within the restrictive environment of the host cell cytosol are poorly understood. In this report, we describe a differential fluorescence-based genetic screen utilizing fluorescence-activated cell sorting (FACS) and high-throughput microscopy to identify L. monocytogenes mutants defective in optimal intracellular replication. Bacteria harboring deletions within the identified gene menD or pepP were defective for growth in primary murine macrophages and plaque formation in monolayers of L2 fibroblasts, thus validating the ability of the screening method to identify intracellular replication-defective mutants. Genetic complementation of the menD and pepP deletion strains rescued the in vitro intracellular infection defects. Furthermore, the menD deletion strain displayed a general extracellular replication defect that could be complemented by growth under anaerobic conditions, while the intracellular growth defect of this strain could be complemented by the addition of exogenous menaquinone. As prior studies have indicated the importance of aerobic metabolism for L. monocytogenes infection, these findings provide further evidence for the importance of menaquinone and aerobic metabolism for L. monocytogenes pathogenesis. Lastly, both the menD and pepP deletion strains were attenuated during in vivo infection of mice. These findings demonstrate that the differential fluorescence-based screening approach provides a powerful tool for the identification of intracellular replication determinants in multiple bacterial systems.