Ouabain-insensitive salt and water movements in duck red cells. I. Kinetics of cation transport under hypertonic conditions

Duck red cells in hypertonic media experience rapid osmotic shrinkage followed by gradual reswelling back toward their original volume. This uptake of salt and water is self limiting and demands a specific ionic composition of the external solution. Although ouabain (10(-4)M) alters the pattern of cation accumulation from predominantly potassium to sodium, it does not affect the rate of the reaction, or the total amount of salt or water taken up. To study the response without the complications of active Na-K transport, ouabain was added to most incubations. All water accumulated by the cells can be accounted for by net salt uptake. Specific external cation requirements for reswelling include: sufficient sodium (more than 23 mM), and elevated potassium (more than 7 mM). In the absence of external potassium cells lose potassium without gaining sodium and continue to shrink instead of reswelling. Adding rubidium to the potassium- free solution promotes an even greater loss of cell potassium, yet causes swelling due to a net uptake of sodium and rubidium followed by chloride. The diuretic furosemide (10(-3)M) inhibits net sodium uptake which depends on potassium (or rubidium), as well as inhibits net sodium uptake which depends on sodium. As a result, cell volume is stabilized in the presence of this drug by inhibition of shrinkage, at low, and of swelling at high external potassium. The response has a high apparent energy of activation (15-20 kcal/mol). We propose that net salt and water movements in hypertonic solutions containing ouabain are mediated by direct coupling or cis-interaction, between sodium and potassium so that the uphill movement of one is driven by the downhill movement of the other in the same direction.     

Tetrapyrrole‐based drought stress signalling

Tetrapyrroles such as chlorophyll and heme play a vital role in primary plant metabolic processes such as photosynthesis and respiration. Over the past decades, extensive genetic and molecular analyses have provided valuable insights into the complex regulatory network of the tetrapyrrole biosynthesis. However, tetrapyrroles are also implicated in abiotic stress tolerance, although the mechanisms are largely unknown. With recent reports demonstrating that modified tetrapyrrole biosynthesis in plants confers wilting avoidance, a component physiological trait to drought tolerance, it is now timely that this pathway be reviewed in the context of drought stress signalling. In this review, the significance of tetrapyrrole biosynthesis under drought stress is addressed, with particular emphasis on the inter‐relationships with major stress signalling cascades driven by reactive oxygen species (ROS) and organellar retrograde signalling. We propose that unlike the chlorophyll branch, the heme branch of the pathway plays a key role in mediating intracellular drought stress signalling and stimulating ROS detoxification under drought stress. Determining how the tetrapyrrole biosynthetic pathway is involved in stress signalling provides an opportunity to identify gene targets for engineering drought‐tolerant crops.     

First direct estimate of the detection probability of bovine tuberculosis in possums by possum transmission

We conducted a field trial to determine whether percutaneous challenge of wild brushtail possums (Trichosurus vulpecula) with virulent Mycobacterium bovis, the causative agent of bovine tuberculosis (TB), resulted in subsequent transmission of M. bovis to other possums. We then used our results to make a preliminary first direct estimate of the ‘TB detection probability’ or transmission rate in wild possums. Seven free-living adult possums in the Rimutaka Forest Park, New Zealand, were captured, GPS-collared, experimentally challenged and then released. Possum populations surrounding the challenged individuals were examined for secondary cases 4–8 months post-challenge. Of 37 possums caught within the home ranges of the challenged individuals, one had confirmed TB. As no TB was detected in 355 other possums examined in this study, or 659 possums in a concurrent study in the immediate vicinity, we conclude that this represents the first documented case of onward transmission of M. bovis from an experimentally challenged possum in the wild. This pilot trial therefore gives some confidence that the new percutaneous challenge model for TB in possums can be used to obtain empirical estimates of TB detection probabilities. The calculated probability of onward transmission (‘detection’) here was just 0.027; however, with the possibility that further transmission events would have occurred from the one challenged possum removed from the trial before it had died, or if our trapping missed secondary cases, this value could be an underestimate. The implications of this low preliminary estimate are discussed.     

Evidence for predatory control of the invasive round goby

We coupled bioenergetics modeling with bottom trawl survey results to evaluate the capacity of piscivorous fish in eastern Lake Erie to exert predatory control of the invading population of round goby Neogobius melanostomus. In the offshore (>20 m deep) waters of eastern Lake Erie, burbot Lota lota is a native top predator, feeding on a suite of prey fishes. The round goby invaded eastern Lake Erie during the late 1990s, and round goby population size increased dramatically during 1999–2004. According to annual bottom trawl survey results, round goby abundance in offshore waters peaked in 2004, but then declined during 2004–2008. Coincidentally, round goby became an important component of burbot diet beginning in 2003. Using bottom trawling and gill netting, we estimated adult burbot abundance and age structure in eastern Lake Erie during 2007. Diet composition and energy density of eastern Lake Erie burbot were also determined during 2007. This information, along with estimates of burbot growth, burbot mortality, burbot water temperature regime, and energy densities of prey fish from the literature, were incorporated into a bioenergetics model application to estimate annual consumption of round goby by the adult burbot population. Results indicated that the adult burbot population in eastern Lake Erie annually consumed 1,361 metric tons of round goby. Based on the results of bottom trawling, we estimated the biomass of yearling and older round goby in offshore waters eastern Lake Erie during 2007–2008 to be 2,232 metric tons. Thus, the adult burbot population was feeding on round goby at an annual rate equal to 61% of the estimated round goby standing stock. We concluded that the burbot population had high potential to exert predatory control on round goby in offshore waters of eastern Lake Erie.     

The involvement of interleukin-1 and interleukin-4 in the response of human annulus fibrosus cells to cyclic tensile strain: an altered mechanotransduction pathway with degeneration

Introduction  

Recent evidence suggests that intervertebral disc (IVD) cells derived from degenerative tissue are unable to respond to physiologically relevant mechanical stimuli in the 'normal' anabolic manner, but instead respond by increasing matrix catabolism. Understanding the nature of the biological processes which allow disc cells to sense and respond to mechanical stimuli (a process termed 'mechanotransduction') is important to ascertain whether these signalling pathways differ with disease. The aim here was to investigate the involvement of interleukin (IL)-1 and IL-4 in the response of annulus fibrosus (AF) cells derived from nondegenerative and degenerative tissue to cyclic tensile strain to determine whether cytokine involvement differed with IVD degeneration.     

Avian biodiversity in the coastal wetlands of the Okahukura Peninsula

The Tapora Landcare Group, operating on the Okahukura Peninsula, has the long-term goal of making this region predator fenced. The aim of this study was to obtain information on the current status of avian biodiversity and the bird community across the band of coastal wetlands on the Okahukura Peninsula. Bird counts were conducted and playback lures used to detect three cryptic wetland species: fernbirds (Bowdleria punctata); spotless crakes (Porzana tabuensis); and banded rails (Gallirallus philippensis). Fernbirds and banded rails were detected at seven of the eight wetland sites sampled whereas spotless crakes were detected at two sites. The native species with the highest relative abundance across the eight sites were silvereyes (Zosterops lateralis) and South Island pied oystercatchers (Haematopus finschi). Changes in avian biodiversity over time in the region can now be monitored, and comprehensive long-term data on the status of avian biodiversity over time obtained.     

Intraspecific transmission of Mycobacterium bovis among penned feral pigs in New Zealand

We investigated intraspecific transmission of Mycobacterium bovis (etiologic agent of bovine tuberculosis [Tb]) among penned pigs (Sus scrofa) in New Zealand. Between 2002 and 2006, we conducted two trials, each involving two 1-ha pens in natural habitat in which uninfected sentinel pigs were kept with wild-caught feral pigs, many of which were infected with M. bovis. The rate of M. bovis transmission to sentinels was used to assess whether intraspecies transmission between live pigs could explain the high levels of infection in the wild population. In trial 1, no new infection was detected in 18 penned sentinels after total exposure of 3.9 yr (2.6 mo/sentinel); three of 11 sentinels in the wild became infected. In trial 2, a more heavily infected source pig population (94% prevalence compared with 42% in the first trial) was used, and one (4%) of 25 penned sentinels became infected. However, 75% of the eight sentinels released to the wild became infected. Combining trials, the difference in apparent annual incidence was significant (mean ± 95% confidence interval = 0.03 ± 0.12 for penned sentinels vs. 1.06 ± 6.74 for released sentinels; t=-3.51, P=0.04). In the pens, infected pigs were kept in contact with susceptible sentinels for 7 yr in total, but only one transmission event was detected. Taking pig longevity into account, the R(0) value (the basic reproductive rate of disease) for intraspecies infection between live pigs seems unlikely to exceed 0.25, even under highly conducive conditions. We suggest that live pigs are unlikely to transmit M. bovis to wild conspecifics often enough to sustain Tb in pigs by this route of transmission alone. The high prevalences seen in some New Zealand feral pig populations are likely to result from transmission from another host or route.     

Optimization of HIV-1 infectivity assays

HIV-1 reporter cell lines are the backbone of diagnostic assays, vaccine and drug development efforts. Performing HIV-1 infection experiments in a T cell background is desirable for many reasons. However, a low susceptibility to infection with primary patient isolates in available reporter T cell lines has limited such efforts. We here demonstrate that optimization of HIV-1 receptor expression and the utilization of serum free medium compositions can increase susceptibility of reporter T cell lines to HIV-1 infection by up to two orders of magnitude.