Continuous growth of vimentin filaments in mouse fibroblasts

We have investigated the dynamics of intermediate filament assembly in vivo by following the fate of heterologous chicken vimentin subunits expressed under the control of an inducible promoter in transfected mouse fibroblasts. Using RNase protection, metabolic protein pulse-chase and immunofluorescence microscopy, we have examined the fate of newly assembled subunits under physiological conditions in situ. Following induction and subsequent removal of inducer, chicken vimentin mRNA had a half-life of approximately 6 h while both chicken and mouse vimentin protein polymer had long half-lives--roughly equivalent to the cell generation time. Moreover, following deinduction, chicken vimentin immunolocalization progressed from a continuous (8-10 h chase) to a discontinuous (> or = 20 h chase) pattern. The continuous chicken vimentin staining reflects the uniform incorporation of chicken vimentin throughout the endogenous mouse vimentin network while the discontinuous or punctate chicken vimentin staining represents short interspersed segments of assembled chicken vimentin superimposed on the endogenous polymer. This punctate staining pattern of chicken vimentin was present throughout the entire array of intermediate filaments, with no bias toward the perinuclear region. These results are consistent with a continuous growth model of intermediate filament assembly, wherein subunit addition occurs at discrete sites located throughout the cytoskeleton

Cell cycle regulation of a Xenopus Wee1-like kinase

Using a polymerase chain reaction-based strategy, we have isolated a gene encoding a Wee1-like kinase from Xenopus eggs. The recombinant Xenopus Wee1 protein efficiently phosphorylates Cdc2 exclusively on Tyr- 15 in a cyclin-dependent manner. The addition of exogenous Wee1 protein to Xenopus cell cycle extracts results in a dose-dependent delay of mitotic initiation that is accompanied by enhanced tyrosine phosphorylation of Cdc2. The activity of the Wee1 protein is highly regulated during the cell cycle: the interphase, underphosphorylated form of Wee1 (68 kDa) phosphorylates Cdc2 very efficiently, whereas the mitotic, hyperphosphorylated version (75 kDa) is weakly active as a Cdc2-specific tyrosine kinase. The down-modulation of Wee1 at mitosis is directly attributable to phosphorylation, since dephosphorylation with protein phosphatase 2A restores its kinase activity. During interphase, the activity of this Wee1 homolog does not vary in response to the presence of unreplicated DNA. The mitosis-specific phosphorylation of Wee1 is due to at least two distinct kinases: the Cdc2 protein and another activity (kinase X) that may correspond to an MPM-2 epitope kinase. These studies indicate that the down-regulation of Wee1-like kinase activity at mitosis is a multistep process that occurs after other biochemical reactions have signaled the successful completion of S phase

Influence of Different Cotton Fruit Sizes on Boll Weevil (Coleoptera: Curculionidae) Oviposition and Survival to Adulthood

Understanding the critical host plant factors that determine oviposition behavior and survival of boll weevil, Anthonomus grandis grandis Boheman, on cotton, Gossypium hirsutum L., is important for developing successful pest management strategies. However, published information is both conflicting and limited regarding how different cotton fruit sizes affect boll weevil oviposition choices and subsequent larval survival to adulthood. Consequently, we used a standard based on fruit size diameter to evaluate boll weevil feeding and oviposition punctures, and survival to adulthood on 10 different cotton fruit sizes: squares of diameter 1.5–2.0 (pinhead), 3.0–3.5 (matchhead), 5–6, 7–8, or 9–10 mm; candle; and bolls of diameter 10–15, 15–20, 20–30, or \u3e30 mm. Oviposition and feeding punctures were significantly affected by cotton fruit size. Females did not oviposit in pinhead squares. The fewest eggs were oviposited in boll sizes \u3e30 mm. The highest number of eggs was recorded in square sizes of 5–6 and 7–8 mm. Boll weevil survival to adulthood was highest on square sizes of 7–8 or 9–10 mm (58.6–59.7%). No survival occurred in matchhead squares or bolls \u3e30 mm. Duration of development was longest on boll sizes of 15–20 and 20–30 mm (18.2–18.8 d). The growth index (percentage immature survival divided by immature developmental time) of female boll weevils was 2.8-fold higher in 7–8- or 9–10-mm diameter squares than in 20–30-mm diameter bolls. This study will improve our capacity to develop methods to predict fruit losses and changes in boll weevil populations in the field, given a starting density of fruit suitable for oviposition, and a corresponding initial population density of weevils

Night-time lighting alters the composition of marine epifaunal communities

Marine benthic communities face multiple anthropogenic pressures that compromise the future of some of the most biodiverse and functionally important ecosystems in the world. Yet one of the pressures these ecosystems face, night-time lighting, remains unstudied. Light is an important cue in guiding the settlement of invertebrate larvae, and altering natural regimes of nocturnal illumination could modify patterns of recruitment among sessile epifauna. We present the first evidence of night-time lighting changing the composition of temperate epifaunal marine invertebrate communities. Illuminating settlement surfaces with white light-emitting diode lighting at night, to levels experienced by these communities locally, both inhibited and encouraged the colonization of 39% of the taxa analysed, including three sessile and two mobile species. Our results indicate that ecological light pollution from coastal development, shipping and offshore infrastructure could be changing the composition of marine epifaunal communities.European Research Council under the European Union's Seventh Framework programme (FP7/2007-2013

Effects of Conventional vs. Conservation Tillage Systems on Population Dynamics of Boll Weevil (Coleoptera: Curculionidae) in Dryland Cotton

Studies were conducted during 2000-2001 to determine the effects of a conservation tillage system in dryland cotton on soil surface temperatures, soil moisture, plant canopy structure, light interception, timing of fruit set, and how these factors affect crop yield and boll weevil, Anthonomus grandis grandis Boheman, populations compared with a conventional moldboard tillage system. Soil moisture at the 10-40 cm depth was 1.6-1.9-fold higher in the conservation tillage treatment than in the conventional tillage treatment throughout the first 90 days of crop growth due to the decreased evaporation from crop residue mulch. The conventional tillage cotton treatment had a greater water stress, causing plants to shed squares and bolls. Cotton plants in the conventional tillage treatment allocated more resources into vegetative growth while the conservation tillage cotton responded by fruiting at a higher rate. At 110 days after planting the conservation tillage cotton had an average height of 42.4 cm per plant versus 63.0 cm in conventional tillage, and the number of leaves per plant was 32.4 versus 51.7, while fruit numbers were 13.0 versus 7.1, respectively. Increased plant height and number of leaves in the conventional tillage provided significantly more light interception and shading of the soil surface. In the conservation tillage cotton, 60.2% of the incoming sunlight reached the soil surface, while the conventional tillage had only 36.2%. Soil temperatures between the rows in conservation tillage cotton were 8-11º C higher than in conventional tillage and significantly influenced boll weevil mortality in infested squares shed from plants. The number of boll weevils per plant was 2.3 to 3.4-fold higher in the conventional tillage compared with the conservation tillage. Trap counts of weevil populations followed a similar trend with 1.6 to 2.8-fold more weevils in the conventional tillage compared to conservation tillage. The mortality of boll weevils in fallen, naturally infested squares, and in cohorts of laboratory-infested squares collected from the middle of the rows was 1.5-1.8-fold higher in the conservation tillage field than in the conventional. Percent punctured squares by boll weevils during the growing season averaged 2.1-fold higher in conventional than in conservation tillage fields

SOST Inhibits Prostate Cancer Invasion.

Inhibitors of Wnt signaling have been shown to be involved in prostate cancer (PC) metastasis; however the role of Sclerostin (Sost) has not yet been explored. Here we show that elevated Wnt signaling derived from Sost deficient osteoblasts promotes PC invasion, while rhSOST has an inhibitory effect. In contrast, rhDKK1 promotes PC elongation and filopodia formation, morphological changes characteristic of an invasive phenotype. Furthermore, rhDKK1 was found to activate canonical Wnt signaling in PC3 cells, suggesting that SOST and DKK1 have opposing roles on Wnt signaling in this context. Gene expression analysis of PC3 cells co-cultured with OBs exhibiting varying amounts of Wnt signaling identified CRIM1 as one of the transcripts upregulated under highly invasive conditions. We found CRIM1 overexpression to also promote cell-invasion. These findings suggest that bone-derived Wnt signaling may enhance PC tropism by promoting CRIM1 expression and facilitating cancer cell invasion and adhesion to bone. We concluded that SOST and DKK1 have opposing effects on PC3 cell invasion and that bone-derived Wnt signaling positively contributes to the invasive phenotypes of PC3 cells by activating CRIM1 expression and facilitating PC-OB physical interaction. As such, we investigated the effects of high concentrations of SOST in vivo. We found that PC3-cells overexpressing SOST injected via the tail vein in NSG mice did not readily metastasize, and those injected intrafemorally had significantly reduced osteolysis, suggesting that targeting the molecular bone environment may influence bone metastatic prognosis in clinical settings

Gauged Fermionic Q-balls

We present a new model for a non-topological soliton (NTS) that contains interacting fermions, scalar particles and a gauge field. Using a variational approach, we estimate the energy of the localized configuration, showing that it can be the lowest energy state of the system for a wide range of parameters.Comment: 5 pages, 2 figures; revised version to appear in Phys. Rev.

Hox-logic of preadaptations for social insect symbiosis in rove beetles

How symbiotic lifestyles evolve from free-living ecologies is poorly understood. Novel traits mediating symbioses may stem from preadaptations: features of free-living ancestors that predispose taxa to engage in nascent interspecies relationships. In Metazoa's largest family, Staphylinidae (rove beetles), the body plan within the subfamily Aleocharinae is preadaptive for symbioses with social insects. Short elytra expose a pliable abdomen that bears targetable glands for host manipulation or chemical defense. The exposed abdomen has also been convergently refashioned into ant- and termite-mimicking shapes in multiple symbiotic lineages. Here we show how this preadaptive anatomy evolved via novel Hox gene functions that remodeled the ancestral coleopteran groundplan. Using the model staphylinid Dalotia coriaria, we abolished activities of the five thoracic and abdominal Hox genes. We show that elytral shortening is a staphylinid-specific property of the Hox-less appendage ground state, which is overridden in the metathorax by Ultrabithorax to promote hind wing expansion. In the exposed abdomen, we present evidence that defensive gland development stems from novel combinatorial outputs of the Abdominal-A and Abdominal-B Hox proteins: in the posterior compartment of tergite VI they specify a chemical gland reservoir, an imaginal disc-like invagination of ectodermal secretory cells; in the anterior compartment of tergite VII Abdominal-A and Abdominal-B specify clusters of classical duct-bearing glands. These distinct gland cell types collectively synthesize a blend of benzoquinone irritants, surfactant esters and alkane solvent, a defensive chemistry, which in symbiotic species has been augmented with specialized volatiles that potently manipulate ant behavior. These results reveal how Hox-controlled body axis modifications caused a convergent trend towards evolving symbiosis in the Metazoa