Characterization and expression analysis of Staphylococcus aureus pathogenicity island 3 - Implications for the evolution of staphylococcal pathogenicity islands

We describe the complete sequence of the 15.9-kb staphylococcal pathogenicity island 3 encoding staphylococcal enterotoxin serotypes B, K, and Q. The island, which meets the generally accepted definition of pathogenicity islands, contains 24 open reading frames potentially encoding proteins of more than 50 amino acids, including an apparently functional integrase. The element is bordered by two 17-bp direct repeats identical to those found flanking staphylococcal pathogenicity island 1. The island has extensive regions of homology to previously described pathogenicity islands, particularly staphylococcal pathogenicity islands 1 and bov. The expression of 22 of the 24 open reading frames contained on staphylococcal pathogenicity island 3 was detected either in vitro during growth in a laboratory medium or serum or in vivo in a rabbit model of toxic shock syndrome using DNA microarrays. The effect of oxygen tension on staphylococcal pathogenicity island 3 gene expression was also examined. By comparison with the known staphylococcal pathogenicity islands in the context of gene expression described here, we propose a model of pathogenicity island origin and evolution involving specialized transduction events and addition, deletion, or recombination of pathogenicity island "modules.

Lunar Atmosphere and Dust Environment Explorer Integration and Test

Integration and test (I&T) of the Lunar Atmosphere and Dust Environment Explorer (LADEE) is presented. A collaborative NASA project between Goddard Space Flight Center and Ames Research Center, LADEE's mission is to explore the low lunar orbit environment and exosphere for constituents. Its instruments include two spectrometers, a dust detector, and a laser communication technology demonstration. Although a relatively low-cost spacecraft, LADEE has I&T requirements typical of most planetary probes, such as prelaunch contamination control, sterilization, and instrument calibration. To lead to a successful mission, I&T at the spacecraft, instrument, and observatory level must include step-by-step and end-to-end functional, environmental, and performance testing. Due to its compressed development schedule, LADEE I&T planning requires adjusting test flows and sequences to account for long-lead critical-path items and limited spares. A protoflight test-level strategy is also baselined. However, the program benefits from having two independent but collaborative teams of engineers, managers, and technicians that have a wealth of flight project experience. This paper summarizes the LADEE I&T planning, flow, facilities, and probe-unique processes. Coordination of requirements and approaches to I&T when multiple organizations are involved is discussed. Also presented are cost-effective approaches to I&T that are transferable to most any spaceflight project I&T program

Unilateral ureteral obstruction impairs renal antioxidant enzyme activation during sodium depletion

Unilateral ureteral obstruction impairs renal antioxidant enzyme activation during sodium depletion.BackgroundObstructive nephropathy leads to progressive renal tubular atrophy and interstitial fibrosis and is associated with sodium wasting and sodium depletion. Renal damage resulting from unilateral ureteral obstruction (UUO) may be aggravated by reactive oxygen species (ROS), which are produced by a variety of processes. Ideally, deleterious effects of ROS are attenuated by antioxidant enzymes, including the superoxide dismutases, glutathione peroxidases, catalase, and glutathione-S-transferases. The general paradigm is that tissue damage occurs when ROS production is greater than the protective capacity of the antioxidant enzymes.MethodsThis study was designed to investigate the response of renal antioxidant enzymes to UUO and sodium depletion. Adult, male Sprague-Dawley rats received normal-sodium or sodium-depleted diets and were subjected to UUO or sham operation. Obstructed (UUO), intact opposite, or sham-operated kidneys were harvested after 14days, and antioxidant enzyme activities were measured in kidney homogenates. Thiobarbituric acid reactive substances were measured in these homogenates at 3 and 14days after UUO or sham operation as an index of ROS production.ResultsRenal interstitial area, a measure of fibrosis, was increased by UUO and was doubled in sodium-depleted animals. Sodium depletion increased manganese superoxide dismutase, glutathione peroxidases, and glutathione-S-transferase activities in sham-operated kidneys but not in UUO kidneys. Relative to intact opposite kidneys, UUO kidneys had reduced activities of catalase, manganese superoxide dismutase, and glutathione-S-transferase in normal-sodium animals and all antioxidant enzymes tested in sodium-depleted animals. Renal thiobarbituric acid reactive substances were increased by three days of UUO and were increased further by 14days of sodium depletion.ConclusionIn summary, sodium depletion increased several renal antioxidant enzymes, consistent with a stress response to increased ROS production. Further, UUO not only reduced antioxidant enzyme activities but also inhibited increases seen with sodium depletion. We conclude that suppression of renal antioxidant enzyme activities by UUO contributes to the progression of renal injury in obstructive nephropathy, a process exacerbated by sodium depletion

Diel CO2 cycles reduce severity of bahavioural abnormalities in coral reef fish under ocean acidification

Elevated CO2 levels associated with ocean acidification (OA) have been shown to alter behavioural responses in coral reef fishes. However, all studies to date have used stable pCO2 treatments, not considering the substantial diel pCO2 variation that occurs in shallow reef habitats. Here, we reared juvenile damselfish, Acanthochromis polyacanthus, and clownfish, Amphiprion percula, at stable and diel cycling pCO2 treatments in two experiments. As expected, absolute lateralization of A. polyacanthus and response to predator cue of Am. percula were negatively affected in fish reared at stable, elevated pCO2 in both experiments. However, diel pCO2 fluctuations reduced the negative effects of OA on behaviour. Importantly, in experiment two, behavioural abnormalities that were present in fish reared at stable 750 μatm CO2 were largely absent in fish reared at 750 ± 300 μatm CO2. Overall, we show that diel pCO2 cycles can substantially reduce the severity of behavioural abnormalities caused by elevated CO2. Thus, past studies may have over-estimated the impacts of OA on the behavioural performance of coral reef fishes. Furthermore, our results suggest that diel pCO2 cycles will delay the onset of behavioural abnormalities in natural populations

Drug-tolerant persister cancer cells are vulnerable to GPX4 inhibition.

Acquired drug resistance prevents cancer therapies from achieving stable and complete responses. Emerging evidence implicates a key role for non-mutational drug resistance mechanisms underlying the survival of residual cancer 'persister' cells. The persister cell pool constitutes a reservoir from which drug-resistant tumours may emerge. Targeting persister cells therefore presents a therapeutic opportunity to impede tumour relapse. We previously found that cancer cells in a high mesenchymal therapy-resistant cell state are dependent on the lipid hydroperoxidase GPX4 for survival. Here we show that a similar therapy-resistant cell state underlies the behaviour of persister cells derived from a wide range of cancers and drug treatments. Consequently, we demonstrate that persister cells acquire a dependency on GPX4. Loss of GPX4 function results in selective persister cell ferroptotic death in vitro and prevents tumour relapse in mice. These findings suggest that targeting of GPX4 may represent a therapeutic strategy to prevent acquired drug resistance

Informing Management of Henrys Lake, Idaho, using an Integrated Catch-at-Age Model

Henrys Lake, Idaho, supports a popular fishery for Yellowstone Cutthroat Trout Oncorhynchus clarkii bouvieri and Yellowstone Cutthroat Trout × Rainbow Trout O. mykiss hybrids. A majority of the adult population of fish in Henrys Lake are of hatchery origin that were stocked as fingerlings. The fishery is closed to angling during the late winter and spring months, but fisheries managers are considering opening the fishery year-round with catch-and-release- only regulations or with a two-fish bag limit during the extended season. However, there is concern that the proposed management actions may negatively affect the current fishery. Therefore, we developed an integrated catch-atage model to estimate population parameters for trout in Henrys Lake and used a simulation model to evaluate alternative management actions. Results of this study suggest that catch and release of both Yellowstone Cutthroat Trout and hybrids would increase and that abundance of trout in the spring (i.e., the start of the traditional season) would decrease under both proposed bag limits. Losses in abundance can be mitigated by stocking additional fish as long as no more than approximately 1,520,000 Yellowstone Cutthroat Trout are stocked annually. If catch-and-release-only regulations are implemented during the newly proposed season, total harvest is expected to decrease compared to the current fishery due to additional catch-and-release mortality. Ultimately, managers will need to prioritize harvest or catch-and-release opportunity, both of which provide additional utility to anglers, when choosing how to proceed with bag limit regulations

Performance and Life Tests of a Regenerative Blower for EVA Suit Ventilation

Ventilation fans for future space suits must meet demanding performance specifications, satisfy stringent safety requirements for operation in an oxygen atmosphere, and be able to increase output to operate in buddy mode. A regenerative blower is an attractive choice due to its ability to meet these requirements at low operating speed. This paper describes progress in the development and testing of a regenerative blower designed to meet requirements for ventilation subsystems in future space suits. The blower includes a custom-designed motor that has significantly improved its efficiency. We have measured the blower s head/flow performance and power consumption under conditions that simulate both the normal and buddy mode operating points. We have operated the blower for TBD hours and demonstrated safe operation in an oxygen test loop at prototypical pressures. We also demonstrated operation with simulated lunar dust

The zebrafish xenograft platform-A novel tool for modeling KSHV-associated diseases

Kaposi\u27s sarcoma associated-herpesvirus (KSHV, also known as human herpesvirus-8) is a gammaherpesvirus that establishes life-long infection in human B lymphocytes. KSHV infection is typically asymptomatic, but immunosuppression can predispose KSHV-infected individuals to primary effusion lymphoma (PEL); a malignancy driven by aberrant proliferation of latently infected B lymphocytes, and supported by pro-inflammatory cytokines and angiogenic factors produced by cells that succumb to lytic viral replication. Here, we report the development of the firs

Long-Wavelength Instability in Marangoni Convection

Our experiments in thin liquid layers (approximately 0.1 mm thick) heated from below reveal a well-defined long-wavelength instability: at a critical temperature difference across the layer, the depth of the layer in the center of the cell spontaneously decreases until the liquid-air interface ruptures and a dry spot forms. The onset of this critical instability occurs at a temperature difference across the liquid layer that is 35% smaller than that predicted in earlier theoretical studies of a single layer model. Our analysis of a two-layer model yields predictions in accord with the observations for liquid layer depths greater than or equal to 0.15 mm, but for smaller depths there is an increasing difference between our predictions and observations (the difference is 25% for a layer 0.06 mm thick). In microgravity environments the long-wavelength instability observed in our terrestrial experiments is expected to replace cellular convection as the primary instability in thick as well as thin liquid layers heated quasistatically from below

Onset of hexagons in surface-tension-driven Benard convection

High resolution laboratory experiments with large aspect ratio are being conducted for thin fluid layers heated from below and bounded from above by a free surface. The fluid depths are chosen sufficiently small (less than 0.06 cm) so that surface tension is the dominant driving mechanisms; the Rayleigh number is less than 5 for the results reported here. Shadowgraph visualization reveals that the primary instability leading to hexagons is slightly hysteretic (approximately 1 percent). Preliminary measurements of the convection amplitude using infrared imaging are also presented