miR-675 mediates downregulation of Twist1 and Rb in AFP-secreting hepatocellular carcinoma.

BACKGROUND: Alpha-fetoprotein (AFP)-secreting hepatocellular carcinomas (HCC) represent a genetically distinct subset of tumors often associated with a worse prognosis. However, the molecular mechanisms that underlie these phenotypic differences remain poorly understood. METHODS: HCC tumor samples from 27 patients were profiled using the Affymetrix 133 Plus 2.0 GeneChips. GeneGO Metacore software was used to identify altered biologic pathways. Expression validation was confirmed by RT-PCR. Manipulation of miR-675 by overexpression and antagomir-mediated knockdown was carried out with subsequent evaluation of effects on cell behavior by cell cycle, proliferation, invasion, and growth in soft agar assays. RESULTS: We identified a strong relationship between primary tumor H19 gene expression and elevated serum AFP. H19 has recently been identified to encode microRNA-675 (miR-675), and we confirmed the relationship in an independent sample of patients. Pathway analyses of the effect of miR-675 overexpression in hepatoma cells revealed a predominant upregulation of cell adhesion and cell cycle initiation pathways. We have demonstrated that miR-675 mediates increases in proliferation and an accumulation of cells with tetraploid DNA content associated with a repression of Rb. We also demonstrated that overexpression of miR-675 alters cellular morphology, reduces invasive potential, and increases anchorage-independent growth capacity. These findings are consistent with a mesenchymal-to-epithelial transition, associated with a reduction in the expression of the key EMT mediator, Twist1. CONCLUSIONS: Expression of the miR-675 in hepatocellular carcinoma links a dramatic upregulation of proliferative and growth capacity with inhibition of motility in HCC cells.

A new context-based approach to assess marine mammal behavioral responses to anthropogenic sounds.

Acute effects of anthropogenic sounds on marine mammals, such as from military sonars, energy development, and offshore construction, have received considerable international attention from scientists, regulators, and industry. Moreover, there has been increasing recognition and concern about the potential chronic effects of human activities (e.g., shipping). It has been demonstrated that increases in human activity and background noise can alter habitats of marine animals and potentially mask communications for species that rely on sound to mate, feed, avoid predators, and navigate. Without exception, regulatory agencies required to assess and manage the effects of noise on marine mammals have addressed only the acute effects of noise on hearing and behavior. Furthermore, they have relied on a single exposure metric to assess acute effects: the absolute sound level received by the animal. There is compelling evidence that factors other than received sound level, including the activity state of animals exposed to different sounds, the nature and novelty of a sound, and spatial relations between sound source and receiving animals (i.e., the exposure context) strongly affect the probability of a behavioral response. A more comprehensive assessment method is needed that accounts for the fact that multiple contextual factors can affect how animals respond to both acute and chronic noise. We propose a three-part approach. The first includes measurement and evaluation of context-based behavioral responses of marine mammals exposed to various sounds. The second includes new assessment metrics that emphasize relative sound levels (i.e., ratio of signal to background noise and level above hearing threshold). The third considers the effects of chronic and acute noise exposure. All three aspects of sound exposure (context, relative sound level, and chronic noise) mediate behavioral response, and we suggest they be integrated into ecosystem-level management and the spatial planning of human offshore activities.

The Southern Ocean Exchange: porous boundaries between humpback whale breeding populations in southern polar waters.

Humpback whales (Megaptera novaeangliae) are a cosmopolitan species and perform long annual migrations between low-latitude breeding areas and high-latitude feeding areas. Their breeding populations appear to be spatially and genetically segregated due to long-term, maternally inherited fidelity to natal breeding areas. In the Southern Hemisphere, some humpback whale breeding populations mix in Southern Ocean waters in summer, but very little movement between Pacific and Atlantic waters has been identified to date, suggesting these waters constituted an oceanic boundary between genetically distinct populations. Here, we present new evidence of summer co-occurrence in the West Antarctic Peninsula feeding area of two recovering humpback whale breeding populations from the Atlantic (Brazil) and Pacific (Central and South America). As humpback whale populations recover, observations like this point to the need to revise our perceptions of boundaries between stocks, particularly on high latitude feeding grounds. We suggest that this "Southern Ocean Exchange" may become more frequent as populations recover from commercial whaling and climate change modifies environmental dynamics and humpback whale prey availability.

The economics of overexploitation.

The general economic analysis of a biological resource presented in this article suggests that overexploitation in the physical sense of reduced productivity may result from not one, but two social conditions: common-property competitive exploitation on the one hand, and private-property maximization of profits on the other. For populations that are economically valuable but possess low reproductive capacities, either condition may lead even to the extinction of the population. In view of the likelihood of private firms adopting high rates of discount, the conservation of renewable resources would appear to require continual public surveillance and control of the physical yield and the condition of the stocks.

Sound Playback Experiments with Southern Right Whales (Eubalaena australis).

A variety of sound recordings were played to southern right whales. Whales approached the loudspeaker and made frequent sounds in response to recordings of other southern right whales, but swam away and made relatively few sounds in response to playbacks of water noise, 200-hertz tones, and humpback whale sounds. Thus it appears that southern right whales can differentiate between con-specific sounds and other sounds.

Management of multispecies fisheries.

With the overexploitation of many conventional fish stcocks, and growing interest in harvesting new kinds of food from the sea, there is increasing need for managers of fisheries to take account of interactions among species. In particular, as Antarctic krill-fishing industries grow, there is a need to agree upon sound principles for managing the Southern Ocean ecosystem. Using simple models, we discuss the way multispecies food webs respond to the harvesting of species at differrent trophic levels. These biological and economic insights are applied to a discussion of fisheries in the Southern Otean and the North Sea and to enunciate some for harvesting in multispecies systems.

Relationship between canine transthoracic impedance and defibrillation threshold. Evidence for current-based defibrillation.

The electrical parameter used to define defibrillation strength is energy. Peak current, however, may more accurately reflect the field quantities (i.e., electric field strength and current density) that mediate defibrillation and therefore should be a better clinical descriptor of threshold than energy. Though transthoracic impedance is a major determinant of energy-based threshold and is sensitive to operator-dependent changes in impedance (electrode-subject interface), an ideal threshold descriptor should be invariant with respect to these changes in impedance. We therefore compared the relative invariance of energy- and current-based thresholds when transthoracic impedance was altered by one of two methods: (a) change in electrode size (protocol A) or (b) change in electrode force (protocol B). In protocol A, impedance was altered in each dog by a mean of 95%. Energy thresholds determined at both low and high impedance were 44 +/- 21 J (mean +/- SD) and 105 +/- 35 J, respectively, P less than 0.0001. In contrast, peak current (A) thresholds were independent of transthoracic impedance, 22 +/- 5 A (low impedance) vs. 24 +/- 6 A (high impedance), P = NS. Energy and current thresholds showed a similar relationship for animals tested in protocol B. Therefore, current-based thresholds, in contrast to energy thresholds are independent of operator-dependent variables of transthoracic impedance and are invariant for a given animal. These results suggest that redefining defibrillation threshold in terms of peak current rather than energy provides a superior method of defibrillation.

Short- and long-term changes in right whale calling behavior: the potential effects of noise on acoustic communication.

The impact of anthropogenic noise on marine mammals has been an area of increasing concern over the past two decades. Most low-frequency anthropogenic noise in the ocean comes from commercial shipping which has contributed to an increase in ocean background noise over the past 150 years. The long-term impacts of these changes on marine mammals are not well understood. This paper describes both short- and long-term behavioral changes in calls produced by the endangered North Atlantic right whale (Eubalaena glacialis) and South Atlantic right whale (Eubalaena australis) in the presence of increased low-frequency noise. Right whales produce calls with a higher average fundamental frequency and they call at a lower rate in high noise conditions, possibly in response to masking from low-frequency noise. The long-term changes have occurred within the known lifespan of individual whales, indicating that a behavioral change, rather than selective pressure, has resulted in the observed differences. This study provides evidence of a behavioral change in sound production of right whales that is correlated with increased noise levels and indicates that right whales may shift call frequency to compensate for increased band-limited background noise.

The effect of dominance on food hoarding: a game theoretical model.

Many food hoarding animals live in small groups structured by rank. The presence of conspecifics in the hoarding area increases the risk of losing stored supplies. The possibility of stealing from others depends on a forager's rank in the group. Highly ranked individuals can steal from subordinates and also protect their own caches. Since storing incurs both costs and benefits, the optimal hoarding investment will differ between individuals of different rank. In a game theoretical model, we investigate how dominant and subordinate individuals should optimize their hoarding effort. Our model imagines animals that are large-scale hoarders in autumn and dependent on stored supplies for winter survival. Many examples can be found in the bird families Paridae and Corvidae, but the model can be used for any hoarding species that forage in groups. Predictions from the model are as follows: First, subordinates should store more than dominants, but in a predictable environment, this difference will decrease as the environment gets harsher. Under harsh conditions, dominants should store almost as much as subordinates and, later, spend almost as much time retrieving their own caches as subordinates. Second, if on the other hand, bad winter conditions were not expected when storing, dominants should spend more time pilfering caches from subordinates. Third, in populations that are highly dependent on stored supplies, dominants should store relatively more than in populations that are less dependent on stored supplies. Fourth, harsher environments will favor hoarding. And finally, if dominant individuals store, it implies that hoarders have a selfish recovery advantage over conspecific pilferers.

Accelerating Scientific Discovery Through Computation and Visualization.

The rate of scientific discovery can be accelerated through computation and visualization. This acceleration results from the synergy of expertise, computing tools, and hardware for enabling high-performance computation, information science, and visualization that is provided by a team of computation and visualization scientists collaborating in a peer-to-peer effort with the research scientists. In the context of this discussion, high performance refers to capabilities beyond the current state of the art in desktop computing. To be effective in this arena, a team comprising a critical mass of talent, parallel computing techniques, visualization algorithms, advanced visualization hardware, and a recurring investment is required to stay beyond the desktop capabilities. This article describes, through examples, how the Scientific Applications and Visualization Group (SAVG) at NIST has utilized high performance parallel computing and visualization to accelerate condensate modeling, (2) fluid flow in porous materials and in other complex geometries, (3) flows in suspensions, (4) x-ray absorption, (5) dielectric breakdown modeling, and (6) dendritic growth in alloys.

Current problems in the management of marine fisheries.

The public perception of fisheries is that they are in crisis and have been for some time. Numerous scientific and popular articles have pointed to the failures of fisheries management that have caused this crisis. These are widely accepted to be overcapacity in fishing fleets, a failure to take the ecosystem effects of fishing into account, and a failure to enforce unpalatable but necessary reductions in fishing effort on fishing fleets and communities. However, the claims of some analysts that there is an inevitable decline in the status of fisheries is, we believe, incorrect. There have been successes in fisheries management, and we argue that the tools for appropriate management exist. Unfortunately, they have not been implemented widely. Our analysis suggests that management authorities need to develop legally enforceable and tested harvest strategies, coupled with appropriate rights-based incentives to the fishing community, for the future of fisheries to be better than their past.

Underwater, low-frequency noise in a coastal sea turtle habitat.

Underwater sound was recorded in one of the major coastal foraging areas for juvenile sea turtles in the Peconic Bay Estuary system in Long Island, New York. The recording season of the underwater environment coincided with the sea turtle activity season in an inshore area where there is considerable boating and recreational activity, especially during the summer between Independence Day and Labor Day. Within the range of sea turtle hearing, average noise pressure reached 110 dB during periods of high human activity and diminished proportionally, down to 80 dB, with decreasing human presence. Therefore, during much of the season when sea turtles are actively foraging in New York waters, their coastal habitats are flooded with underwater noise. During the period of highest human activity, average noise pressures within the range of frequencies heard by sea turtles were greater by over two orders of magnitude (26 dB) than during the lowest period of human activity. Sea turtles undoubtedly are exposed to high levels of noise, most of which is anthropogenic. Results suggest that continued exposure to existing high levels of pervasive anthropogenic noise in vital sea turtle habitats and any increase in noise could affect sea turtle behavior and ecology.

Dynamic models of behavior: An extension of life history theory.

Life history theory and behavioral ecology, two branches in the study of adaptation, have relied extensively on mathematical models, but have tended to employ different types of models, and different currencies of fitness. Recently, a new approach based on dynamic, state variable models has been increasingly applied to the study of behavioral adaptations. In fact, this approach amounts to a unification of life history theory and behavioral ecology, to the extent that the line separating the two fields is virtually obliterated. Dynamic models (usually solved by computer) can yield both general principles and testable, quantitative or qualitative predictions about specific behavioral and life history phenomena.

Isotope shifts of C I spectral lines and their application to radioactive dating by laser-assisted mass spectrometry.

We have calculated isotope shifts for several one- and two-photon transitions in neutral carbon. The results provide a unified interpretation of existing experimental data, and they demonstrate the applicability of a new method of ultrasensitive isotope trace analysis to carbon.

A delayed-recruitment model of population dynamics, with an application to baleen whale populations.

This paper studies the delay equation chik + 1 = lambda chik + F(chik - beta), which has been employed as a model of baleen whale population dynamics. The two main questions discussed are (a) stability of equilibria, and (b) optimal exploitation policies.

Behavioral responses of humpback whales (Megaptera novaeangliae) to full-scale ATOC signals.

Loud (195 dB re 1 microPa at 1 m) 75-Hz signals were broadcast with an ATOC projector to measure ocean temperature. Respiratory and movement behaviors of humpback whales off North Kauai, Hawaii, were examined for potential changes in response to these transmissions and to vessels. Few vessel effects were observed, but there were fewer vessels operating during this study than in previous years. No overt responses to ATOC were observed for received levels of 98-109 dB re 1 microPa. An analysis of covariance, using the no-sound behavioral rate as a covariate to control for interpod variation, found that the distance and time between successive surfacings of humpbacks increased slightly with an increase in estimated received ATOC sound level. These responses are very similar to those observed in response to scaled-amplitude playbacks of ATOC signals [Frankel and Clark, Can. J. Zool. 76, 521-535 (1998)]. These similar results were obtained with different sound projectors, in different years and locations, and at different ranges creating a different sound field. The repeatability of the findings for these two different studies indicates that these effects, while small, are robust. This suggests that at least for the ATOC signal, the received sound level is a good predictor of response.

Identification of the central regulatory segment of plasmid R6K complexed with the membranes of Escherichia coli.

Understanding the mechanisms which control replication of chromosomes with multiple origins of replication have been the subject of many investigations. The plasmid, R6K, has three origins of replication, alpha, beta, and gamma. In vivo, alpha and beta are utilized with equal frequencies while the gamma origin is rarely used in the parental plasmid, or remains silent in several miniplasmids. Hence, in the present study, the multiple origin plasmid, R6K, was utilized as a model system to investigate DNA replication. A 1.6 kb PCR (polymerase chain reaction) amplified region of R6K containing the central regulatory segment (CRS) was examined to determine if membrane complexing is required for maintenance. Crude membrane extracts from exponentially growing cultures were prepared and sedimented through 30-50% (w/v) linear sucrose gradients. Fractions collected were probed using quantitative PCR which revealed two fractions containing CRS DNA. These complexes were localized in a region between the inner and outer membranes of Escherichia coli.

Population dynamics of sexual and resource competition.

Interspecific competition is usually thought of in terms of resource competition. Closely related species, however, may often compete also for a "sexual niche," a sexual resource. Both sexual and resource competition are likely to affect the dynamics of closely related sympatric species. Here, a combined model of sexual and resource competition is developed and analyzed in terms of phase planes. Population dynamics involving sexual competition alone are characterized by competitive exclusion and dependence on relative initial population size of the competing species. This dependence on initial population size is due to the nonlinear equilibrium isoclines of sexual competition, which contrast with the linear isoclines of the Lotka-Volterra competition equations. Dependence on initial population size is stronger when sexual competition is severe and/or when growth rates of the two species are low. Additional dynamic possibilities arise under combined sexual and resource competition. If the sexual competition is strong, competitive exclusion is the only outcome. But if sexual competition is weak, and if stable coexistence would occur without sexual competition, then both stable coexistence and competitive exclusion may occur simultaneously. A species that is relatively rare initially may be excluded, but when both species are initially of comparable abundance both will persist. Adding sexual competition to resource competition thus introduces additional uncertainty concerning the outcome of competition between two sympatric species.