Scientific results from Gulf of Mexico Gas Hydrates Joint Industry Project Leg 1 drilling : introduction and overview

This paper is not subject to U.S. copyright. The definitive version was published in Marine and Petroleum Geology 25 (2008): 819-829, doi:10.1016/j.marpetgeo.2008.02.007.The Gulf of Mexico Gas Hydrates Joint Industry Project (JIP) is a consortium of production and service companies and some government agencies formed to address the challenges that gas hydrates pose for deepwater exploration and production. In partnership with the U.S. Department of Energy and with scientific assistance from the U.S. Geological Survey and academic partners, the JIP has focused on studies to assess hazards associated with drilling the fine-grained, hydrate-bearing sediments that dominate much of the shallow subseafloor in the deepwater (>500 m) Gulf of Mexico. In preparation for an initial drilling, logging, and coring program, the JIP sponsored a multi-year research effort that included: (a) the development of borehole stability models for hydrate-bearing sediments; (b) exhaustive laboratory measurements of the physical properties of hydrate-bearing sediments; (c) refinement of new techniques for processing industry-standard 3-D seismic data to constrain gas hydrate saturations; and (d) construction of instrumentation to measure the physical properties of sediment cores that had never been removed from in situ hydrostatic pressure conditions. Following review of potential drilling sites, the JIP launched a 35-day expedition in Spring 2005 to acquire well logs and sediment cores at sites in Atwater Valley lease blocks 13/14 and Keathley Canyon lease block 151 in the northern Gulf of Mexico minibasin province. The Keathley Canyon site has a bottom simulating reflection at not, vert, ~ 392 m below the seafloor, while the Atwater Valley location is characterized by seafloor mounds with an underlying upwarped seismic reflection consistent with upward fluid migration and possible shoaling of the base of the gas hydrate stability (BGHS). No gas hydrate was recovered at the drill sites, but logging data, and to some extent cores, suggest the occurrence of gas hydrate in inferred coarser-grained beds and fractures, particularly between 220 and 330 m below the seafloor at the Keathley Canyon site. This paper provides an overview of the results of the initial phases of the JIP work and introduces the 15 papers that make up this special volume on the scientific results related to the 2005 logging and drilling expedition.Supported by the U.S. Department of Energy, under award DE-FC26-01NT4133

Social transmission of Pavlovian fear: fear-conditioning by-proxy in related female rats

Pairing a previously neutral conditioned stimulus (CS; e.g., a tone) to an aversive unconditioned stimulus (US; e.g., a foot-shock) leads to associative learning such that the tone alone will elicit a conditioned response (e.g., freezing). Individuals can also acquire fear from a social context, such as through observing the fear expression of a conspecific. In the current study, we examined the influence of kinship/familiarity on social transmission of fear in female rats. Rats were housed in triads with either sisters or non-related females. One rat from each cage was fear conditioned to a tone CS+ shock US. On day two, the conditioned rat was returned to the chamber accompanied by one of her cage mates. Both rats were allowed to behave freely, while the tone was played in the absence of the foot-shock. The previously untrained rat is referred to as the fear-conditioned by-proxy (FCbP) animal, as she would freeze based on observations of her cage-mate’s response rather than due to direct personal experience with the foot-shock. The third rat served as a cage-mate control. The third day, long-term memory tests to the CS were performed. Consistent with our previous application of this paradigm in male rats (Bruchey et al. in Behav Brain Res 214(1):80–84, 2010), our results revealed that social interactions between the fear conditioned and FCbP rats on day two contribute to freezing displayed by the FCbP rats on day three. In this experiment, prosocial behavior occurring at the termination of the cue on day two was significantly greater between sisters than their non-sister counterparts, and this behavior resulted in increased freezing on day three. Our results suggest that familiarity and/or kinship influences the social transmission of fear in female rats

Endoplasmic reticulum stress disrupts placental morphogenesis: implications for human intrauterine growth restriction.

We recently reported the first evidence of placental endoplasmic reticulum (ER) stress in the pathophysiology of human intrauterine growth restriction. Here, we used a mouse model to investigate potential underlying mechanisms. Eif2s1(tm1RjK) mice, in which Ser51 of eukaryotic initiation factor 2 subunit alpha (eIF2α) is mutated, display a 30% increase in basal translation. In Eif2s1(tm1RjK) placentas, we observed increased ER stress and anomalous accumulation of glycoproteins in the endocrine junctional zone (Jz), but not in the labyrinthine zone where physiological exchange occurs. Placental and fetal weights were reduced by 15% (97 mg to 82 mg, p < 0.001) and 20% (1009 mg to 798 mg, p < 0.001), respectively. To investigate whether ER stress affects bioactivity of secreted proteins, mouse embryonic fibroblasts (MEFs) were derived from Eif2s1(tm1RjK) mutants. These MEFs exhibited ER stress, grew 50% slower, and showed reduced Akt-mTOR signalling compared to wild-type cells. Conditioned medium (CM) derived from Eif2s1(tm1RjK) MEFs failed to maintain trophoblast stem cells in a progenitor state, but the effect could be rescued by exogenous application of FGF4 and heparin. In addition, ER stress promoted accumulation of pro-Igf2 with altered glycosylation in the CM without affecting cellular levels, indicating that the protein failed to be processed after release. Igf2 is the major growth factor for placental development; indeed, activity in the Pdk1-Akt-mTOR pathways was decreased in Eif2s1(tm1RjK) placentas, indicating loss of Igf2 signalling. Furthermore, we observed premature differentiation of trophoblast progenitors at E9.5 in mutant placentas, consistent with the in vitro results and with the disproportionate development of the labyrinth and Jz seen in placentas at E18.5. Similar disproportion has been reported in the Igf2-null mouse. These results demonstrate that ER stress adversely affects placental development, and that modulation of post-translational processing, and hence bioactivity, of secreted growth factors contributes to this effect. Placental dysmorphogenesis potentially affects fetal growth through reduced exchange capacity

\u3ci\u3eStaphylococcus aureus\u3c/i\u3e Hyaluronidase Is a CodY-Regulated Virulence Factor

Staphylococcus aureus is a Gram-positive pathogen that causes a diverse range of bacterial infections. Invasive S. aureus strains secrete an extensive arsenal of hemolysins, immunomodulators, and exoenzymes to cause disease. Our studies have focused on the secreted enzyme hyaluronidase (HysA), which cleaves the hyaluronic acid polymer at the β-1,4 glycosidic bond. In the study described in this report, we have investigated the regulation and contribution of this enzyme to S. aureus pathogenesis. Using the Nebraska Transposon Mutant Library (NTML), we identified eight insertions that modulate extracellular levels of HysA activity. Insertions in the sigB operon, as well as in genes encoding the global regulators SarA and CodY, significantly increased HysA protein levels and activity. By altering the availability of branched-chain amino acids, we further demonstrated CodY-dependent repression of HysA activity. Additionally, through mutation of the CodY binding box upstream of hysA, the repression of HysA production was lost, suggesting that CodY is a direct repressor of hysA expression. To determine whether HysA is a virulence factor, a ΔhysA mutant of a community-associated methicillin-resistant S. aureus (CA-MRSA) USA300 strain was constructed and found to be attenuated in a neutropenic, murine model of pulmonary infection. Mice infected with this mutant strain exhibited a 4-log-unit reduction in bacterial burden in their lungs, as well as reduced lung pathology and increased levels of pulmonary hyaluronic acid, compared to mice infected with the wild-type, parent strain. Taken together, these results indicate that S. aureus hyaluronidase is a CodY-regulated virulence factor

Electrophilic PPARγ Ligands Attenuate IL-1β and Silica-Induced Inflammatory Mediator Production in Human Lung Fibroblasts via a PPARγ-Independent Mechanism

Acute and chronic lung inflammation is associated with numerous important disease pathologies including asthma, chronic obstructive pulmonary disease and silicosis. Lung fibroblasts are a novel and important target of anti-inflammatory therapy, as they orchestrate, respond to, and amplify inflammatory cascades and are the key cell in the pathogenesis of lung fibrosis. Peroxisome proliferator-activated receptor gamma (PPARγ) ligands are small molecules that induce anti-inflammatory responses in a variety of tissues. Here, we report for the first time that PPARγ ligands have potent anti-inflammatory effects on human lung fibroblasts. 2-cyano-3, 12-dioxoolean-1, 9-dien-28-oic acid (CDDO) and 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) inhibit production of the inflammatory mediators interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), COX-2, and prostaglandin (PG)E2 in primary human lung fibroblasts stimulated with either IL-1β or silica. The anti-inflammatory properties of these molecules are not blocked by the PPARγ antagonist GW9662 and thus are largely PPARγ independent. However, they are dependent on the presence of an electrophilic carbon. CDDO and 15d-PGJ2, but not rosiglitazone, inhibited NF-κB activity. These results demonstrate that CDDO and 15d-PGJ2 are potent attenuators of proinflammatory responses in lung fibroblasts and suggest that these molecules should be explored as the basis for novel, targeted anti-inflammatory therapies in the lung and other organs

The relative susceptibility of grapevine rootstocks to black foot disease is dependent on inoculum pressure

Black foot disease of grapevines is a major economic issue for the viticulture industry worldwide. The disease is mainly associated with a complex of pathogen species within the genera Dactylonectria and Ilyonectria. The susceptibility of six grapevine rootstock cultivars to black foot disease under field conditions was assessed. Callused rootstocks of 101-14, 5C, 420A, Riparia Gloire, Schwarzmann and 3309C were planted into soil containing low natural pathogen populations or inoculated with isolates representing the species diversity in New Zealand. Disease incidence, disease severity and dry weight accumulation were assessed after 8 months of growth. Root and shoot dry weights were not significantly affected by inoculation treatment, but differed among rootstock cultivars, with cultivar 420A having the lowest root and shoot dry weight, cultivar 3309C having the largest shoot dry weight and cultivar 5C the largest root dry weight. The relative susceptibility of rootstocks differed significantly depending on whether they were grown under low natural inoculum pressure or a higher pressure in artificially inoculated soil. Schwarzmann and Riparia Gloire rootstock cultivars were the least susceptible under natural low inoculum pressure, but were the most susceptible in inoculated soil. In contrast, 5C was one of the most susceptible under low inoculum levels but was the least susceptible under high pathogen pressure. The result of the study indicate that black foot pathogen inoculum levels in soil affect the relative susceptibility of grapevine rootstocks to infection, and may have implications for the selection of rootstocks for planting

Studies of the dynamics of nuclear clustering in human syncytiotrophoblast

Syncytial nuclear aggregates (SNAs), clusters of nuclei in the syncytiotrophoblast of the human placenta, are increased as gestation advances and in pregnancy pathologies. The origins of increased SNAs are unclear; however, a better appreciation of the mechanism may give insight into placental ageing and factors underpinning dysfunction. We developed three models to investigate whether SNA formation results from a dynamic process of nuclear movement and to generate alternative hypotheses. SNA count and size were measured in placental explants cultured over 16 days and particles released into culture medium were quantified. Primary trophoblasts were cultured for 6 days. Explants and trophoblasts were cultured with and without cytoskeletal inhibitors. An in silico model was developed to examine the effects of modulating nuclear behaviour on clustering. In explants, neither median SNA number (108 SNA/mm(2) villous area) nor size (283 μm(2)) changed over time. Subcellular particles from conditioned culture medium showed a wide range of sizes that overlapped with those of SNAs. Nuclei in primary trophoblasts did not change position relative to other nuclei; apparent movement was associated with positional changes of the syncytial cell membrane. In both models, SNAs and nuclear clusters were stable despite pharmacological disruption of cytoskeletal activity. In silico, increased nuclear movement, adhesiveness and sites of cytotrophoblast fusion were related to nuclear clustering. The prominence of SNAs in pregnancy disorders may not result from an active process involving cytoskeleton-mediated rearrangement of syncytial nuclei. Further insights into the mechanism(s) of SNA formation will aid understanding of their increased presence in pregnancy pathologies