To assess the potential impact of the Deepwater Horizon oil spill on offshore ecosystems, 11 sites hosting deep-water coral communities were examined 3 to 4 mo after the well was capped. Healthy coral communities were observed at all sites >20 km from the Macondo well, including seven sites previously visited in September 2009, where the corals and communities appeared unchanged. However, at one site 11 km southwest of the Macondo well, coral colonies presented widespread signs of stress, including varying degrees of tissue loss, sclerite enlargement, excess mucous production, bleached commensal ophiuroids, and covering by brown flocculent material (floc). On the basis of these criteria the level of impact to individual colonies was ranked from 0 (least impact) to 4 (greatest impact). Of the 43 corals imaged at that site, 46% exhibited evidence of impact on more than half of the colony, whereas nearly a quarter of all of the corals showed impact to >90% of the colony. Additionally, 53% of these corals' ophiuroid associates displayed abnormal color and/or attachment posture. Analysis of hopanoid petroleum biomarkers isolated from the floc provides strong evidence that this material contained oil from the Macondo well. The presence of recently damaged and deceased corals beneath the path of a previously documented plume emanating from the Macondo well provides compelling evidence that the oil impacted deep-water ecosystems. Our findings underscore the unprecedented nature of the spill in terms of its magnitude, release at depth, and impact to deep-water ecosystems.

Emerging evidence indicates that membrane lipids regulate protein networking by directly interacting with protein-interaction domains (PIDs). As a pilot study to identify and functionally annodate lipid-binding PIDs on a genomic scale, we performed experimental and computational studies of PDZ domains. Characterization of 70 PDZ domains showed that ∼40% had submicromolar membrane affinity. Using a computational model built from these data, we predicted the membrane-binding properties of 2,000 PDZ domains from 20 species. The accuracy of the prediction was experimentally validated for 26 PDZ domains. We also subdivided lipid-binding PDZ domains into three classes based on the interplay between membrane- and protein-binding sites. For different classes of PDZ domains, lipid binding regulates their protein interactions by different mechanisms. Functional studies of a PDZ domain protein, rhophilin 2, suggest that all classes of lipid-binding PDZ domains serve as genuine dual-specificity modules regulating protein interactions at the membrane under physiological conditions.

Prostaglandins (PGs) are emerging as important immune mediators. Since our first report on the expression of prostacyclin synthase in the germinal centers, we have investigated production mechanisms and biological functions of PG using human follicular dendritic cell (FDC)-like cells. In the previous report, we observed that TGF-β enhances PG production, and IL-4 prevents this upregulation. To elucidate the inhibitory mechanism of IL-4, its effects on the key enzyme leading to PG production were analyzed in this study. IL-4 but not IL-10 inhibited TGF-β-induced COX-2 expression both at mRNA and protein levels. Next the early signaling molecules of IL-4 were identified by siRNA technology. IL-4 induced tyrosine phosphorylation of STAT1, 3, and 6, but only JAK1-STAT6 pathway was responsible for the prevention of COX-2 augmentation and PG production. Phosphorylated STAT6 accumulated in the nucleus rapidly upon IL-4 addition, and the complete inhibition of COX-2 upregulation required 24h of pretreatment with IL-4, implying that newly transcribed molecules mediate the inhibitory signals downstream of STAT6. Interestingly, unphosphorylated STAT6 proteins were constitutively expressed in the nucleus, and depletion of STAT6 impaired background level expression of COX-2 and PGs. Our results highlight the crucial roles of TGF-β and IL-4 in the regulation of PG production, which lead us to suggest that T cells play an important role in FDC production of PGs.

Phage display screening with a combinatorial library was used to identify M13 type bacteriophage that express peptides with selective binding to organic crystals of thiamethoxam. The six most strongly binding phage exhibit at least 1000 times the binding affinity of wild-type M13 and express heptapeptide sequences that are rich in hydrophobic, hydrogen bonding amino acids and proline. Among the peptide sequences identified, M13 displaying the pIII domain heptapeptide ASTLPKA exhibits the strongest binding to thiamethoxam in competitive binding assays. Electron and confocal microscopy confirm the specific binding affinity of ASTLPKA to thiamethoxam. Using atomic force microscope (AFM) probes functionalized with ASTLPKA expressing phage, the average adhesion force between the bacteriophage and a thiamethoxam surface is 1.47 ± 0.80 nN, while the adhesion force of wild-type M13KE phage is 0.18 ± 0.07 nN. Such strongly binding bacteriophage could be used to modify the surface chemistry of thiamethoxam crystals and other organic solids with a high degree of specificity.

In this paper, we present a new segmentation method using the level set framework for medical volume images. The method was implemented using the surface evolution principle based on the geometric deformable model and the level set theory. And, the speed function in the level set approach consists of a hybrid combination of three integral measures derived from the calculus of variation principle. The terms are defined as robust alignment, active region, and smoothing. These terms can help to obtain the precise surface of the target object and prevent the boundary leakage problem. The proposed method has been tested on synthetic and various medical volume images with normal tissue and tumor regions in order to evaluate its performance on visual and quantitative data. The quantitative validation of the proposed segmentation is shown with higher Jaccard's measure score (72.52%-94.17%) and lower Hausdorff distance (1.2654mm-3.1527mm) than the other methods such as mean speed (67.67%-93.36% and 1.3361mm-3.4463mm), mean-variance speed (63.44%-94.72% and 1.3361mm-3.4616mm), and edge-based speed (0.76%-42.44% and 3.8010mm-6.5389mm). The experimental results confirm that the effectiveness and performance of our method is excellent compared with traditional approaches.

The effect of rf wave frequencies on the production of H(-) ion is investigated in a transformer coupled plasma H(-) ion source at Seoul National University. A Langmuir probe is installed to measure the plasma density and temperature, and these plasma parameters are correlated to the extracted H(-) beam currents at various frequencies. The Langmuir probe is also used to measure the density of H(-) ions at the ion source by generating photodetachment with an Nd:YAG laser. The extracted H(-) currents decrease to a minimum value until 13 MHz and then, increase as the driving frequency increases from 13 MHz while the relative H(-) population measured by photodetachment monotonically decreases as the driving rf frequency increases from 11 MHz to 15 MHz. A potential well formed at the extraction region at high frequencies of more than 13 MHz is considered responsible for the increased H(-) beam extraction even with a lower photodetachment signal. The variation in the driving rf frequency not only affects the density and temperature of the plasma but also modifies the plasma potential with the existence of a filtering magnetic field and consequently, influences the extracted H(-) current through the extraction as well as formation of H(-) ions.

Studies suggest that Hsf4 expression correlates with its role in cell growth and differentiation. However, Hsf4's role in tumorigenesis in vivo remains unexplored. In this report, we provide evidence that absence of the Hsf4 gene suppresses evolution of spontaneous tumors arising in p53 or Arf -deficient mice. Furthermore, deletion of hsf4 alters the tumor spectrum by significantly inhibiting development of lymphomas that are normally observed in the majority of mice lacking p53 or Arf tumor suppressor genes. Using mouse embryo fibroblasts deficient in the hsf4 gene we have found that these cells exhibit reduced proliferation that is associated with induction of senescence and senescence-associated-β-galactosidase (SA-β-gal). Cellular senescence in hsf4-deficient cells is associated with the increased expression of the CDK inhibitors, p21 and p27 proteins. Consistent with the cellular senescence observed in vitro, specific normal tissues of hsf4-/- mice and tumors that arose in mice deficient in both hsf4 and p53 genes exhibit increased SA-β-gal activity and elevated levels of p27 compared to wild-type mice. These results suggest that hsf4 deletion-induced senescence is also present in vivo. Our results therefore indicate that Hsf4 is involved in modulation of cellular senescence, which can be exploited during cancer therapy.

BACKGROUND Prostaglandins (PGs) play pathogenic and protective roles in inflammatory diseases. The novel concept of PGs as immune modulators is being documented by several investigators. By establishing an in vitro experimental model containing human follicular dendritic cell-like cells, HK cells, we reported that HK cells produce prostaglandin E(2)(PGE(2)) and prostaglandin I(2)(PGI(2)) and that these PGs regulate biological functions of T and B cells. METHODS To investigate the respective contribution of cyclooxygenase-1 (COX-1) and COX-2 to PGE(2) and PGI(2) production in HK cells, we performed siRNA technology to knock down COX enzymes and examined the effect on PG production. RESULTS Both PGE(2) and PGI(2) productions were almost completely inhibited by the depletion of COX-2. In contrast, COX-1 knockdown did not significantly affect PG production induced by lipopolysaccharide (LPS). CONCLUSION The current results suggest that mPGES-1 and PGIS are coupled with COX-2 but not with COX-1 in human follicular dendritic cell (FDC) and may help understand the potential effects of selective COX inhibitors on the humoral immunity.

ABSTRACT: Study Design. A clinical retrospective study.Objective. To investigate the clinical outcomes of surgery for cervical spine metastasis and identify factors that affecting survival and neurological result.Summary of Background Data. Metastatic disease in the cervical spine is known to be a poorer prognosis than in thoracic and lumbar regions. Few reports focused on cervical spine metastasis are available.Methods. A retrospective analysis of medical records and radiological data was performed on 46 patients who underwent surgical treatment due to cervical spine metastasis from 2000 to 2010. The incidence of cervical metastasis, overall survival (OS), progression free survival (PFS), neurological and pain outcomes were analyzed. In addition, factors affecting survival, local recurrence, and neurologic and pain outcomes among the 46 study subjects were analyzed. These factors included; age, sex, primary tumor growth rate, preoperative disease-progression status (expressed with Tomita score), irradiation, timing of irradiation, postoperative adjuvant therapy, time of diagnosis of spinal metastasis, cord compression or foraminal invasion on magnetic resonance image (MRI), preoperative neurological status and preoperative pain level.Results. The incidence of cervical metastasis was 17.3%. Mean postoperative OS was 16.89 months, the recurrence rate was 39.1%, and mean PFS was 11.82 months. Factors related to prolonged survival were; slow primary tumor growth, low Tomita score, irradiation of the lesion, and postoperative adjuvant therapy. Postoperative adjuvant therapy was also found to be effective for preventing recurrence. Patients with high preoperative Japanese Orthopedic Association Score (JOAS) achieved better neurological outcomes. Foraminal invasion was found to be negatively correlated with postoperative pain outcome.Conclusion. Surgical management for subaxial cervical spinal metastasis was found to be effective in terms of neurologic recovery and pain control. Furthermore, surgical treatment plus adjuvant therapy was found to achieve sufficient local control during postoperative follow-up.

Signaling lipids control many of the most important biological pathways, typically by recruiting cognate protein binding targets to cell surfaces, thereby regulating both their function and subcellular localization. A critical family of signaling lipids is that of the phosphatidylinositol polyphosphates (PIP(n)s), which is composed of seven isomers that vary based on phosphorylation pattern. A key protein that is activated upon PIP(n) binding is Akt, which then plays important roles in regulating the cell cycle, and is thus aberrant in disease. Characterization of protein-PIP(n) binding interactions is hindered by the complexity of the membrane environment and of the PIP(n) structures. Herein, we describe two rapid assays of use for characterizing protein-PIP(n) binding interactions. First, a microplate-based binding assay was devised to characterize the binding of effectors to immobilized synthetic PIP(n) headgroup-biotin conjugates corresponding to all seven isomers. The assay was implemented for simultaneous analysis of Akt-PH domain, indicating PI(3,4,5)P(3) and PI(3,4)P(2) as the primary ligands. In addition, density-dependant studies indicated that the amount of ligand immobilized on the surface affected the amplitude of protein binding, but not the affinity, for Akt-PH. Since the PIP(n) ligand motifs used in this analysis lack the membrane environment and glycerolipid backbone, yet still exhibit high-affinity protein binding, these results narrow down the structural requirements for Akt recognition. Additionally, binding detection was also achieved through microarray analysis via the robotic pin printing of ligands onto glass slides in a miniaturized format. Here, fluorescence-based detection provided sensitive detection of binding using minimal amounts of materials. Due to their high-throughput and versatile attributes, these assays provide invaluable tools for probing and perturbing protein-membrane binding interactions.