Recruitment of Ascophyllum Nodosum: Wave Action as a Source of Mortality

The brown alga Ascophyllum nodosum is a dominant rocky intertidal organism throughout much of the North Atlantic Ocean, yet its inability to colonize exposed or denuded shores is well recognized. Our experimental data show that wave action is a major source of mortality to recently settled zygotes. Artificially recruited zygotes consistently exhibited a Type IV survivorship curve in the presence of moving water. As few as 10, but often only 1 relatively low energy wave removed 85 to 99% of recently settled zygotes. Increasing the setting time for attachment of zygotes (prior to disturbance from water movement) had a positive effect on survival. However, survival was significantly lower at high densities, and decreased at long (24 h) setting times, probably as a result of bacteria on the surface of zygotes. Spatial refuges provided significant protection from gentle water movement but relatively little protection from waves.

Recruitment Habitats and Nursery Grounds of the American Lobster Homarus Americanus: A Demographic Bottleneck?

We have identified benthic recruitment habitats and nursery grounds of the American lobster Homarus americanus Milne Edwards in the coastal Gulf of Maine, USA, by systematically censusing subtidal sediment, cobble, and ledge substrata. We distinguish lobsters between settlement size (5 mm carapace length (CL) to ca 40 mm CL as the 'early benthic phase' (EBP) because they are ecologically and behaviorally distinct from larger lobsters. EBP lobsters are cryptic and apparently restricted to shelter-providing habitats (primarily cobble substratum) in coastal Gulf of Maine. In these habitats we found average population densities of EBP lobsters as high as 6.9 m-2. EBP lobsters were virtually absent from ledge and sedimentary substrata devoid of vegetation although larger lobsters are commonly found there. It is possible that the requirement for shelter-providing substrata by this life phase creates a natural demographic 'bottleneck' to benthic recruitment for the species. Prime cobble recruitment habitat is relatively rare and comprises ca 11 % of the 60.2 km of shoreline at our study area in midcoast Maine. If this low availability of cobble exists throughout the Gulf of Maine, as other studies indicate, it could limit lobster production potential. We verified the geographic extent of recruitment to cobble habitats censused in 3 of 4 regions spanning ca 300 km of the coastal Gulf of Maine (from Nahant, Massachusetts to Swans Island, Maine). Early benthic phase lobsters were absent from cobble censused in the northeastern extreme of our survey (Swans Island). This pattern is consistent with earlier speculation that relatively cool water temperatures may limit larval settlement in this region.

Association of ALLHAT investigator characteristics with participant recruitment

Clinical trials are often not successful because of the inability to recruit a sufficient number of patients. The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT), the largest antihypertensive trial ever conducted, provided highly generalized results and successful recruitment of over 42,000 participants. The overall purpose of this study was to examine the association of investigator characteristics with anti-hypertensive (AHT) participant recruitment in ALLHAT. This secondary data analyses collected data from the ALLHAT investigator profile survey and related investigator characteristics to recruitment success. The sample size was 502 investigators, with recruitment data from 37,947AHT participants. Recruitment was dichotomized by categorizing all sites with recruitment numbers at or above the overall median recruitment number of 46 as "Successful Recruitment". Frequency distributions and univariate and multivariate logistic regression were conducted. When adjusting for all other factors, Hispanic ethnicity, suburban setting, Department of Veterans Affairs Medical Centers (VAMC) site type, number of clinical site staff working on the trial, study coordinator hours per week, medical conference sessions attended, the investigator's primary goal and the likelihood that a physician will convince a patient to continue on randomized treatment, have significant impacts on the recruitment success of ALLHAT investigators. Most of the ALLHAT investigators described their primary commitment as being towards their patients and not to scientific knowledge alone. However, investigators that distinguished themselves as leaders in research had greater recruitment success than investigators who were leaders in clinical practice. ALLHAT was a highly successful trial that proved that community based cardiovascular trials can be implemented on a large scale. Exploring characteristics of ALLHAT investigators provides data that can be generalized to sponsors, sites, and others interested in maximizing clinical trial recruitment numbers. Future studies should further evaluate investigator and study coordinator factors that impact cardiovascular clinical trial recruitment success.^

Binding of the HIV-1 gp120 -V3 to cellular glycosphingolipids is necessary for CXCR4 recruitment and infection

Infection by human immunodeficiency virus type 1 (HIV-1) is a multi-step process, and detailed analyses of the various events critical for productive infection are necessary to clearly understanding the infection process and identifying novel targets for therapeutic interventions. Evidence from this study reveals binding of the viral envelope protein to host cell glycosphingolipids (GSLs) as a novel event necessary for the orderly progression of the host cell-entry and productive infection by HIV-1. Data obtained from co-immunoprecipitation analyses and confocal microscopy showed that the ability of viral envelope to interact with the co-receptor CXCR4 and productive infection of HIV-1 were inhibited in cells rendered GSL-deficient, while both these activities were restored after reconstitution of the cells with specific GSLs like GM3. Furthermore, evidence was obtained using peptide-inhibitors of HIV-1 infection to show that binding of a specific region within the V3-loop of the envelope protein gp120 to the host cell GSLs is the trigger necessary for the CD4-bound gp120 to recruit the CXCR4 co-receptor. Infection-inhibitory activity of the V3 peptides was compromised in GSL-deficient cells, but could be restored by reconstitution of GSLs. Based on these findings, a revised model for HIV-1 infection is proposed that accounts for the established interactions between the viral envelope and host cell receptors while enumerating the importance of the new findings that fill the gap in the current knowledge of the sequential events for the HIV-1 entry. According to this model, post-CD4 binding of the HIV-1 envelope surface protein gp120 to host cell GSLs, mediated by the gp120-V3 region, enables formation of the gp120-CD4-GSL-CXCR4 immune-complex and productive infection. The identification of cellular GSLs as an additional class of co-factors necessary for HIV-1 infection is important for enhancing the basic knowledge of the HIV-1 entry that can be exploited for developing novel antiviral therapeutic strategies. ^

Ocean acidification impairs vermetid reef recruitment

Vermetids form reefs in sub-tropical and warm-temperate waters that protect coasts from erosion, regulate sediment transport and accumulation, serve as carbon sinks and provide habitat for other species. The gastropods that form these reefs brood encapsulated larvae; they are threatened by rapid environmental changes since their ability to disperse is very limited. We used transplant experiments along a natural CO2 gradient to assess ocean acidification effects on the reef-building gastropod Dendropoma petraeum. We found that although D. petraeum were able to reproduce and brood at elevated levels of CO2, recruitment success was adversely affected. Long-term exposure to acidified conditions predicted for the year 2100 and beyond caused shell dissolution and a significant increase in shell Mg content. Unless CO2 emissions are reduced and conservation measures taken, our results suggest these reefs are in danger of extinction within this century, with significant ecological and socioeconomic ramifications for coastal systems.

Conflict and cognitive control during sentence comprehension: recruitment of a frontal network during the processing of Spanish object-first sentences.

During sentence processing there is a preference to treat the first noun phrase found as the subject and agent, unless marked the other way. This preference would lead to a conflict in thematic role assignment when the syntactic structure conforms to a non-canonical object-before-subject pattern. Left perisylvian and fronto-parietal brain networks have been found to be engaged by increased computational demands during sentence comprehension, while event-reated brain potentials have been used to study the on-line manifestation of these demands. However, evidence regarding the spatiotemporal organization of brain networks in this domain is scarce. In the current study we used Magnetoencephalography to track spatio-temporally brain activity while Spanish speakers were reading subject- and object-first cleft sentences. Both kinds of sentences remained ambiguous between a subject-first or an object-first interpretation up to the appearance of the second argument. Results show the time-modulation of a frontal network at the disambiguation point of object-first sentences. Moreover, the time windows where these effects took place have been previously related to thematic role integration (300–500 ms) and to sentence reanalysis and resolution of conflicts during processing (beyond 500 ms post-stimulus). These results point to frontal cognitive control as a putative key mechanism which may operate when a revision of the sentence structure and meaning is necessary

Can seed production and restricted dispersal limit recruitment in Pinus pinaster Aiton from the Spanish Northern Plateau?

Natural regeneration faces increasing difficulties in dry forests from the Mediterranean basin, including for normally well-regenerating species such as maritime pine (Pinus pinaster Aiton). In this paper, we studied female fertility, seed dispersal and spread rates in P. pinaster from the Spanish Northern Plateau, where natural regeneration failure is a main concern for forest managers. For this purpose we periodically collected data from seed traps and trees located at two core locations across several years. We found significant variation in interannual cone production, with the best seed trees being the same across years. In addition, we found highly skewed distributions of female reproductive effort and large fertility differences across stands located few kilometres away. Annual seed dispersal kernels fitted lognormal or 2Dt models depending on the stand analysed, with median dispersal distances between 14 and 25 m. Kernels fitted for maximum dispersal periods showed an outstanding intraseasonal variation of median dispersal distances, from 10 to 54 m, in association to variable patterns of rainfall and maximum wind speed. The amount of seed produced appeared to be enough to guarantee the natural regeneration of the stands during the typical 20-year regeneration period. Colonisation simulations concluded that Mediterranean maritime pine has a notable dispersion capacity, which is strongly influenced by levels of fecundity and, especially, by the number and frequency of long-distance dispersal events. The latter play a key role in tree dispersion processes through enlarging the occupied area and fostering the invasion of abandoned crop land.

Efficient recruitment of TFIIB and CBP-RNA polymerase II holoenzyme by an interferon-β enhanceosome in vitro

The transcriptional activity of an in vitro assembled human interferon-β gene enhanceosome is highly synergistic. This synergy requires five distinct transcriptional activator proteins (ATF2/c-JUN, interferon regulatory factor 1, and p50/p65 of NF-κB), the high mobility group protein HMG I(Y), and the correct alignment of protein-binding sites on the face of the DNA double helix. Here, we investigate the mechanisms of enhanceosome-dependent transcriptional synergy during preinitiation complex assembly in vitro. We show that the stereospecific assembly of the enhanceosome is critical for the efficient recruitment of TFIIB into a template-committed TFIID-TFIIA-USA (upstream stimulatory activity complex) and for the subsequent recruitment of the RNA polymerase II holoenzyme complex. In addition, we provide evidence that recruitment of the holoenzyme by the enhanceosome is due, at least in part, to interactions between the enhanceosome and the transcriptional coactivator CREB, cAMP responsive element binding protein (CBP). These studies reveal a unique role of enhanceosomes in the cooperative assembly of the transcription machinery on the human interferon-β promoter.

Tissue- and stage-specific Wnt target gene expression is controlled subsequent to β-catenin recruitment to cis-regulatory modules

FUNDING This work was supported by the Biotechnology and Biological Sciences Research Council [BB/I003746/1 to S.H., BB/M001695/1 to S.H and Y.N]

Recruitment of IRAK to the interleukin 1 receptor complex requires interleukin 1 receptor accessory protein

The proinflammatory cytokine interleukin 1 (IL-1) activates the transcription of many genes encoding acute phase and proinflammatory proteins, a function mediated primarily by the transcription factor NF-κB. An early IL-1 signaling event is the recruitment of the Ser/Thr kinase IRAK to the type I IL-1 receptor (IL-1RI). Here we describe the function of a previously identified IL-1 receptor subunit designated IL-1 receptor accessory protein (IL-1RAcP). IL-1 treatment of cells induces the formation of a complex containing both IL-1RI and IL-1RAcP. IRAK is recruited to this complex through its association with IL-1RAcP. Overexpression of an IL-1RAcP mutant lacking its intracellular domain, the IRAK-binding domain, prevented the recruitment of IRAK to the receptor complex and blocked IL-1-induced NF-κB activation.

Neuropeptide release by efficient recruitment of diffusing cytoplasmic secretory vesicles

Neuropeptides are slowly released from a limited pool of secretory vesicles. Despite decades of research, the composition of this pool has remained unknown. Endocrine cell studies support the hypothesis that a population of docked vesicles supports the first minutes of hormone release. However, it has been proposed that mobile cytoplasmic vesicles dominate the releasable neuropeptide pool. Here, to determine the cellular basis of the releasable pool, single green fluorescent protein-labeled secretory vesicles were visualized in neuronal growth cones with the use of an inducible construct or total internal reflection fluorescence microscopy. We report that vesicle movement follows the diffusion equation. Furthermore, rapidly moving secretory vesicles are used more efficiently than stationary vesicles near the plasma membrane to support stimulated release. Thus, randomly moving cytoplasmic vesicles participate in the first minutes of neuropeptide release. Importantly, the preferential recruitment of diffusing cytoplasmic secretory vesicles contributes to the characteristic slow kinetics and limited extent of sustained neuropeptide release.

Recruitment of SMRT/N-CoR-mSin3A-HDAC-repressing complexes is not a general mechanism for BTB/POZ transcriptional repressors: The case of HIC-1 and γFBP-B

Hypermethylated in cancer (HIC-1), a new candidate tumor suppressor gene located in 17p13.3, encodes a protein with five C2H2 zinc fingers and an N-terminal broad complex, tramtrack, and bric à brac/poxviruses and zinc-finger (BTB/POZ) domain found in actin binding proteins or transcriptional regulators involved in chromatin modeling. In the human B cell lymphoma (BCL-6) and promyelocityc leukemia (PLZF) oncoproteins, this domain mediates transcriptional repression through its ability to recruit a silencing mediator of retinoid and thyroid hormone receptor (SMRT)/nuclear receptor corepressor (N-CoR)-mSin3A-histone deacetylase (HDAC) complex, a mechanism shared with numerous transcription factors. HIC-1 appears unique because it contains a 13-aa insertion acquired late in evolution, because it is not found in its avian homologue, γF1-binding protein isoform B (γFBP-B), a transcriptional repressor of the γF-crystallin gene. This insertion, located in a conserved region involved in the dimerization and scaffolding of the BTB/POZ domain, mainly affects slightly the ability of the HIC-1 and γFBP-B BTB/POZ domains to homo- and heterodimerize in vivo, as shown by mammalian two-hybrid experiments. Both the HIC-1 and γFBP-B BTB/POZ domains behave as autonomous transcriptional repression domains. However, in striking contrast with BCL-6 and PLZF, both HIC-1 and γFBP-B similarly fail to interact with members of the HDAC complexes (SMRT/N-CoR, mSin3A or HDAC-1) in vivo and in vitro. In addition, a general and specific inhibitor of HDACs, trichostatin A, did not alleviate the HIC-1- and γFBP-B-mediated transcriptional repression, as previously shown for BCL-6. Taken together, our studies show that the recruitment onto target promoters of an HDAC complex is not a general property of transcriptional repressors containing a conserved BTB/POZ domain.

Plo1 Kinase Recruitment to the Spindle Pole Body and Its Role in Cell Division in Schizosaccharomyces pombe

Polo kinases execute multiple roles during cell division. The fission yeast polo related kinase Plo1 is required to assemble the mitotic spindle, the prophase actin ring that predicts the site for cytokinesis and for septation after the completion of mitosis (Ohkura et al., 1995; Bahler et al., 1998). We show that Plo1 associates with the mitotic but not interphase spindle pole body (SPB). SPB association of Plo1 is the earliest fission yeast mitotic event recorded to date. SPB association is strong from mitotic commitment to early anaphase B, after which the Plo1 signal becomes very weak and finally disappears upon spindle breakdown. SPB association of Plo1 requires mitosis-promoting factor (MPF) activity, whereas its disassociation requires the activity of the anaphase-promoting complex. The stf1.1 mutation bypasses the usual requirement for the MPF activator Cdc25 (Hudson et al., 1990). Significantly, Plo1 associates inappropriately with the interphase SPB of stf1.1 cells. These data are consistent with the emerging theme from many systems that polo kinases participate in the regulation of MPF to determine the timing of commitment to mitosis and may indicate that pole association is a key aspect of Plo1 function. Plo1 does not associate with the SPB when septation is inappropriately driven by deregulation of the Spg1 pathway and remains SPB associated if septation occurs in the presence of a spindle. Thus, neither Plo1 recruitment to nor its departure from the SPB are required for septation; however, overexpression of plo1+ activates the Spg1 pathway and causes transient Cdc7 recruitment to the SPB and multiple rounds of septation.