Limiting Spread of a Unicolonial Invasive Insect and Characterization of Seasonal Patterns of Range Expansion

Limiting dispersal is a fundamental strategy in the control of invasive species, and in certain situations containment of incipient populations may be an important management technique. To test the feasibility of slowing the rapid spread of two Argentine ant (Linepithema humile) supercolonies in Haleakala National Park, Hawaii, we applied ant bait and toxicant within an experimental plot situated along a supercolony boundary. The 120×260 m plot simulated a small section of what could potentially be a 120 m wide treatment encompassing the entire expanding boundaries of both supercolonies. Foraging ant numbers at baited monitoring stations decreased sharply within two weeks after treatment, and ant spread was completely halted within the plot for at least one year. In contrast, an adjacent untreated colony boundary advanced an average of 65.2 m over the course of 1 year. Most of this spread took place in the summer and fall, at the time of highest ant abundance at bait monitoring stations, while no outward dispersal occurred during the spring and early summer. These patterns are consistent with the hypothesis that local budding dispersal in this unicolonial species stems from density dependent pressure rather than inherent founding behavior associated with mating. Based on results from this experiment, we are investigating the effectiveness of annual boundary treatments in slowing the Argentine ant invasion at Haleakala National Park. The goals of this program are to protect populations of native arthropods and to keep options open for eventual attempts at eradication. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular and functional analysis of the ruv region of Escherichia coli K-12 reveals three genes involved in DNA repair and recombination

Summary Recombinant plasmids carrying ruvA, ruvB, or both were constructed and used to investigate the genetic defects in a collection of UV-sensitive ruv mutants. The results revealed that efficient survival of UV-irradiated cells depends on both ruvA and ruvB, and on a third gene, ruvC, located upstream of the ruvAB operon. Southern blotting analysis was used to locate insertions in ruv and to examine putative deletion mutants. Two Tn10 insertions were located to the region encoding ruvA. Since these insertions caused a deficiency in the activities of both ruvA and ruvB, we concluded that they must exert a polar effect on ruvB. Two putative ruv deletion mutants were shown to be the result of deletion-inversion events mediated during imprecise excision of Tn10. The relevant inversion breakpoints in these mutants were located to ruvA and ruvC.     

How I learned to love carrots: the role of the cytoskeleton in shaping plant cells

The carrot cell suspension was originally used because it provided a model system for studying directional cell expansion - a key process in plant morphogenesis. Early immunofluorescence studies of plant microtubules, using these cells, provided hints that the cortical array of microtubules was dynamic and this was later confirmed by microinjection studies on plant epidermal cells. A nonfixation approach for detecting F-actin was then developed on these cells and showed that, unlike animal cells, actin filaments remained associated with the nucleus throughout division and could have a role in aligning the plane of cell division. Currently, we are using detergent-extracted carrot cytoskeletons for isolating microtubule-associated proteins (MAPs). I discuss how MAPs may be involved in the oriented deposition of cellulose in the cell wall.     

Sampling efficacy for the red imported fire ant Solenopsis invicta (Hymenoptera: Formicidae)

Cost-effective detection of invasive ant colonies before establishment in new ranges is imperative for the protection of national borders and reducing their global impact. We examined the sampling efficiency of food-baits and pitfall traps (baited and nonbaited) in detecting isolated red imported fire ant (Solenopsis invicta Buren) nests in multiple environments in Gainesville, FL. Fire ants demonstrated a significantly higher preference for a mixed protein food type (hotdog or ground meat combined with sweet peanut butter) than for the sugar or water baits offered. Foraging distance success was a function of colony size, detection trap used, and surveillance duration. Colony gyne number did not influence detection success. Workers from small nests (0- to 15-cm mound diameter) traveled no >3 m to a food source, whereas large colonies (>30-cm mound diameter) traveled up to 17 m. Baited pitfall traps performed best at detecting incipient ant colonies followed by nonbaited pitfall traps then food baits, whereas food baits performed well when trying to detect large colonies. These results were used to create an interactive model in Microsoft Excel, whereby surveillance managers can alter trap type, density, and duration parameters to estimate the probability of detecting specified or unknown S. invicta colony sizes. This model will support decision makers who need to balance the sampling cost and risk of failure to detect fire ant colonies.     

Single and cell population respiratory oscillations in yeast: a 2-photon scanning laser microscopy study

Two-photon scanning laser and confocal microscopies were used to image metabolic dynamics of single or cell populations of Saccharomyces cerevisiae strain 28033. Autofluorescence of reduced nicotinamide nucleotides, and mitochondrial membrane potential (DeltaPsim), were simultaneously monitored. Spontaneous, large-scale synchronized oscillations of NAD(P)H and DeltaPsim throughout the entire population of yeasts occurred under perfusion with aerated buffer in a continuous single-layered film of organisms. These oscillations stopped in the absence of perfusion and the intracellular NAD(P)H pool became reduced. Individual mitochondria within a single yeast also showed in-phase synchronous responses with the cell population, in both tetramethylrhodamine ethyl ester (or tetramethylrhodamine methyl ester) and autofluorescence. A single, localized, laser flash also triggered mitochondrial oscillations in single cells suggesting that the mitochondrion may behave as an autonomous oscillator. We conclude that spontaneous oscillations of S. cerevisiae mitochondrial redox states and DeltaPsim occur within individual yeasts, and synchrony of populations of organisms indicates the operation of an efficient system of cell-cell interaction to produce concerted metabolic multicellular behaviour on the minute time scale in both cases.     

Diurnal Oscillations of Gas Production and Effluxes (CO2 and CH4) in Cores from a Peat Bog

Peat cores (15 cm diam X 30 cm deep) from Ellergower Moss, New Galloway, Scotland were kept and monitored at constant temperature (10 ± 0.1ºC) for gas production using a 1.6 mm diam stainless steel probe fitted with a membrane inlet and connected to a quadrupole mass spectrometer. In the headspace, O₂, CO₂ and CH₄ (measured at m/z values 32, 44 and 15 respectively) showed diurnal fluctuations in low-intensity natural daylight and under a light-dark (LD, 12:12) regime. Over the first few cycles O₂ and CO₂ increased together in the dark and decreased in the light, whereas CH₄ showed variations in antiphase with the other two gases. CO₂ and CH₄ also showed diurnal oscillations at 15 cm depth, but these decreased together in the light whereas argon (m/z = 40) was not varying. A highly-damped free-run of the oscillations in gas concentrations at 15cm depth was evident for only 3 cycles in complete darkness and at constant temperature. This might suggest desynchronization between individual plants with different free-running periods. A hydrocarbon signal (m/z = 26) at 15 cm depth also showed diurnal cycles but out of phase with CO 2 and CH 4 . We postulate a circadian control of microbiological activities imposed by the vascular plants (Carex, Eriophorum, Molinia, Calluna, Erica). Under natural conditions the pronounced temperature sensitivity of CO₂ and CH₄ emission results in entrainment to daily temperature cycles. The amplitudes of the rhythms are greatest when temperature and light intensity changes are most pronounced, i.e. when the fluctuations in environmental factors are most potent as synchronizers (zeitgebers) and as masking factors.     

The reliability of the ankle-brachial index in the Atherosclerosis Risk in Communities (ARIC) study and the NHLBI Family Heart Study (FHS)

Background

Organ transplantation is presently often the only available option to repair a damaged heart. As heart donors are scarce, engineering of cardiac grafts from autologous skeletal myoblasts is a promising novel therapeutic strategy. The functionality of skeletal muscle cells in the heart milieu is, however, limited because of their inability to integrate electrically and mechanically into the myocardium. Therefore, in pursuit of improved cardiac integration of skeletal muscle grafts we sought to modify primary skeletal myoblasts by overexpression of the main gap-junctional protein connexin 43 and to study electrical coupling of connexin 43 overexpressing myoblasts to cardiac myocytes in vitro.

Methods

To create an efficient means for overexpression of connexin 43 in skeletal myoblasts we constructed a bicistronic retroviral vector MLV-CX43-EGFP expressing the human connexin 43 cDNA and the marker EGFP gene. This vector was employed to transduce primary rat skeletal myoblasts in optimised conditions involving a concomitant use of the retrovirus immobilising protein RetroNectin^® and the polycation transduction enhancer Transfectam^®. The EGFP-positive transduced cells were then enriched by flow cytometry.

Results

More than four-fold overexpression of connexin 43 in the transduced skeletal myoblasts, compared with non-transduced cells, was shown by Western blotting. Functionality of the overexpressed connexin 43 was demonstrated by microinjection of a fluorescent dye showing enhanced gap-junctional intercellular transfer in connexin 43 transduced myoblasts compared with transfer in non-transduced myoblasts. Rat cardiac myocytes were cultured in multielectrode array culture dishes together with connexin 43/EGFP transduced skeletal myoblasts, control non-transduced skeletal myoblasts or alone. Extracellular field action potential activation rates in the co-cultures of connexin 43 transduced skeletal myoblasts with cardiac myocytes were significantly higher than in the co-cultures of non-transduced skeletal myoblasts with cardiac myocytes and similar to the rates in pure cultures of cardiac myocytes.

Conclusion

The observed elevated field action potential activation rate in the co-cultures of cardiac myocytes with connexin 43 transduced skeletal myoblasts indicates enhanced cell-to-cell electrical coupling due to overexpression of connexin 43 in skeletal myoblasts. This study suggests that retroviral connexin 43 transduction can be employed to augment engineering of the electrocompetent cardiac grafts from patients' own skeletal myoblasts.