Occupancy and abundance of wintering birds in a dynamic agricultural landscape

Assessing wildlife management action requires monitoring populations, and abundance often is the parameter monitored. Recent methodological advances have enabled estimation of mean abundance within a habitat using presence–absence or count data obtained via repeated visits to a sample of sites. These methods assume populations are closed and intuitively assume habitats within sites change little during a field season. However, many habitats are highly variable over short periods. We developed a variation of existing occupancy and abundance models that allows for extreme spatio-temporal differences in habitat, and resulting changes in wildlife abundance, among sites and among visits to a site within a field season. We conducted our study in sugarcane habitat within the Everglades Agricultural Area southeast of Lake Okeechobee in south Florida. We counted wintering birds, primarily passerines, within 245 sites usually 5 times at each site during December 2006–March 2007. We estimated occupancy and mean abundance of birds in 6 vegetation states during the sugarcane harvest and allowed these parameters to vary temporally or spatially within a vegetation state. Occupancy and mean abundance of the common yellowthroat (Geothlypis trichas) was affected by structure of sugarcane and uncultivated edge vegetation (occupancy = 1.00 [ = 0.96–1.00] and mean abundance = 7.9 [ = 3.2–19.5] in tall sugarcane with tall edge vegetation versus 0.20 [ = 0.04–0.71] and 0.22 [ = 0.04–1.2], respectively, in short sugarcane with short edge vegetation in one half of the study area). Occupancy and mean abundance of palm warblers (Dendroica palmarum) were constant (occupancy = 1.00, = 0.69–1.00; mean abundance = 18, = 1–270). Our model may enable wildlife managers to assess rigorously effects of future edge habitat management on avian distribution and abundance within agricultural landscapes during winter or the breeding season. The model may also help wildlife managers make similar management decisions involving other dynamic habitats such as wetlands, prairies, and even forested areas if forest management or fires occur during the field season. © 2011 The Wildlife Society.     

Using multiple methods to assess detection probabilities of forest-floor wildlife

Many previous comparisons of multiple sampling methods have assumed that detection probabilities for each method are either constant or equal to one. We used 4 sampling methods to estimate detection probabilities for forest-floor dwelling amphibians, reptiles, and small mammals. We investigated associations between seasonality and precipitation on species detection and explored sample design tradeoffs for future studies. Although we captured 25 species, we could reliably detect (detection probability >0.15) only northern short-tailed shrews (Blarina brevicauda) and pygmy and masked shrews (Sorex spp.) using drift fences and red-backed salamanders (Plethodon cinereus) using visual encounter surveys (VES). The use of multiple sampling methods improved detection probabilities for only red-backed salamanders ( = 0.32, 95% CI: 0.24–0.38, = 0.38, 95% CI: 0.32−0.44). Parameter estimates indicated detection of both shrew species was positively related to increased precipitation. Detection probabilities for pygmy and masked shrews and red-backed salamanders were positively and negatively associated with date, respectively. Our power analysis revealed that sampling during rain events increased the power of detecting a change in sorid occupancy by ≥40% (α = 0.05). Our results demonstrate the need to incorporate species detectability when comparing the effectiveness of different trapping methodologies. Furthermore, our study highlights the utility of power analyses for exploring study design tradeoffs for research and monitoring programs. © 2011 The Wildlife Society.     

Estimating occupancy to monitor northern goshawk in the central Rocky Mountains

Designing monitoring programs to evaluate trends in low-density wildlife species at regional scales is challenging given difficulties detecting uncommon organisms distributed in potential habitats over large spatial extents. The northern goshawk (Accipiter gentilis) has been petitioned for listing under the Endangered Species Act and the review of the petition indicated a need for information on population trend. To evaluate trends in goshawk populations, the U.S. Forest Service developed the Northern Goshawk Bioregional Monitoring Design to estimate goshawk occupancy over broad spatial extents. We adapted and implemented this design to approximately 30,600 km² of 88,128 km² of National Forest System lands in the Forest Service Rocky Mountain Region, including portions of Colorado, Wyoming, and South Dakota. We developed a stratified random design to monitor goshawk occupancy in sampling units, defined by primary and secondary habitat quality as well as accessibility. To define habitat quality, we examined a time series for 58 previously located nesting territories. Using logistic regression, we found that the dominant conifer species and status of aspen in postfledging zones best characterized high-quality goshawk nesting habitat. We applied model results to stratify 4,445 sampling units based on habitat quality and further stratified sampling units based on accessibility into easy and difficult access categories. We conducted field sampling during the goshawk breeding season in the summer of 2006 to estimate detection probabilities and occupancy rates. Within our sampling frame, we sampled 51 sampling units and estimated goshawk occupancy of 0.329 (95% CI: 0.213–0.445). Occupancy within primary strata (high quality) sampling units was 0.811 (SE = 0.113), whereas occupancy in secondary strata (lower quality) sampling units was 0.124 (SE = 0.067). Future implementation of this monitoring program can achieve 0.8 power to detect 30–40% declines in with 140 sampling units. Our implementation of a stratified sampling design to monitor occupancy of goshawks at a region-wide scale reduced the number of sampling units in each administrative unit and focused our efforts on those areas most likely to have goshawks. © 2011 The Wildlife Society.     

The role of predator removal,density-dependence,and environmental factors on mallard duckling survival in North Dakota

Duckling survival is an important component of mallard (Anas platyrhynchos) recruitment and population growth, yet many factors regulating duckling survival are poorly understood. We investigated factors affecting mallard duckling survival in the drift prairie of northeastern North Dakota, 2006–2007. Mammalian meso-predators were removed by trapping on 4 92.3 km² study sites and another 4 study sites served as controls. We monitored 169 broods using telemetry and periodic resighting, and we modeled cumulative survival to 30 days of age using the nest survival module in Program MARK. Duckling survival was not affected by predator removal ( , 85% CI: 0.182–0.234; , 85% CI: 0.155–0.211) and was only weakly negatively correlated with duckling density. Duckling survival was higher in 2007 ( , 85% CI: 0.193–0.355) than 2006 ( , 85% CI: 0.084–0.252) and increased with total seasonal and semipermanent wetland area and declined with perennial cover in the surrounding landscape. Broods that hatched earlier in the season (especially in 2006) and ducklings that were heavier at hatch also had higher survival. Our estimates of duckling survival are among the lowest reported for mallards and contradict previous research in Saskatchewan that found predator removal increased duckling survival. However, our results are consistent with other studies suggesting that earlier hatch date, increased wetland availability, and better duckling condition lead to increased survival. Management actions that increase wetland density, improve nest success early in the season, and potentially target brood-specific predators such as mink (Neovison vison) would likely lead to higher duckling survival. © 2011 The Wildlife Society.     

Declining old‐forest species as a legacy of large trees lost

Aim

Global declines in large old trees from selective logging have degraded old‐forest ecosystems, which could lead to delayed declines or losses of old‐forest‐associated wildlife populations (i.e., extinction debt). We applied the declining population paradigm and explored potential evidence for extinction debt in an old‐forest dependent species across landscapes with different histories of large tree logging.

Location

Montane forests of the Sierra Nevada, California, USA.

Methods

We tested hypotheses about the influence of forest structure on territory extinction dynamics of the spotted owl (Strix occidentalis) using detection/non‐detection data from 1993 to 2011 across two land tenures: national forests, which experienced extensive large tree logging over the past century, and national parks, which did not.

Results

Large tree/high canopy cover forest was the best predictor of extinction rates and explained 26%–77% of model deviance. Owl territories with more large tree/high canopy cover forest had lower extinction rates, and this forest type was ~4 times more prevalent within owl territories in national parks ( = 19% of territory) than national forests ( = 4% of territory). As such, predicted extinction probability for an average owl territory was ~2.5 times greater in national forests than national parks, where occupancy was declining () and stable (), respectively. Large tree/high canopy cover forest remained consistently low, but did not decline, during the study period on national forests while owl declines were ongoing—an observation consistent with an extinction debt.

Main conclusions

In identifying a linkage between large trees and spotted owl dynamics at a regional scale, we provide evidence suggesting past logging of large old trees may have contributed to contemporary declines in an old‐forest species. Strengthening protections for remaining large old trees and promoting their recruitment in the future will be critical for biodiversity conservation in the world's forests.

     

Abundance trends of American martens in Michigan based on statistical population reconstruction

Estimating the dynamics of furbearer populations is challenging because their elusive behavior and low densities make observations difficult. Statistical population reconstruction is a flexible approach to demographic assessment for harvested populations, but the technique has not been applied to furbearers. We extended this approach to furbearers and analyzed 8 yr of age-at-harvest data for American marten (Martes americana) in the Upper Peninsula of Michigan. Marten abundance estimates showed a general downward trend from an estimate of = 1,733.3 animals in 2000 to = 1,163.9 in 2007. The harvest probability of martens increased nearly 5-fold from 0.0542 in 2000 to 0.2637 in 2007, which corresponded to a 5-fold increase in trap-nights. Continued monitoring of martens in the Upper Peninsula, Michigan, and a reassessment of current harvest regulations are necessary given the estimated decreases. Moreover, we do not encourage the use of harvest indices as the sole technique to assess the status and trends of marten and fisher populations. Auxiliary studies in the Upper Peninsula, Michigan, will allow for continued use and improvement in the application of these models. © 2011 The Wildlife Society.     

Reduced oxidative activity of circulating neutrophils in patients after myocardial infarction

Circulating neutrophils isolated from patients 3–4 h after a myocardial infarction produced less $ {\rm O}\frac{ \cdot }{{\rm 2}} $ compared with controls, when stimulated with phorbol myrystate acetate or formyl-methionine-leucine-phenylalanine. Three days after the infraction the $ {\rm O}\frac{ \cdot }{{\rm 2}} $ generation elicited by both stimuli further decreased markedly. Seven and 15 days after infarction the $ {\rm O}\frac{ \cdot }{{\rm 2}} $ stimulated production was only slightly lower than or similar to the control values. The neutrophils of infarcted patients showed an augmented latency period before $ {\rm O}\frac{ \cdot }{{\rm 2}} $ production compared with controls in response to exogenous stimuli, particularly three days after infarction. Electron microscopy revealed that the neutrophils isolated from the infarcted patients displayed signs of cell exhaustion with few alterations of the plasma membranes when stimulated with phorbol ester. In contrast, control neutrophils displayed alterations of the plasma membranes characteristic of active neutrophils. The results of this study indicate that the circulating neutrophils appear exhausted and functionally inhibited immediately after myocardial infarction.     

Site Occupancy Dynamics of Northern Spotted Owls in the Eastern Cascades,Washington, USA, 1990–2003

Abstract: Northern spotted owls (Strix occidentalis caurina) have received intense research and management interest since their listing as a threatened species by the United States Fish and Wildlife Service in 1990. Several spotted owl (Strix occidentalis) response variables have been examined in various investigations, but recent advances in statistical modeling permit evaluations of temporal and spatial variability in site occupancy, local-extinction, and colonization probabilities while incorporating imperfect detection probabilities. Following recent work by other researchers on site occupancy dynamics of spotted owls in Oregon, USA, we evaluated temporal variability of detection, occupancy, local-extinction, and colonization probabilities for spotted owls, as well as potential influences of barred owl (Strix varia) presence on these parameters. We used spotted owl survey data collected from 1990 to 2003 on a study area in the eastern Cascades Mountains, Washington, USA, to compare competing occupancy models from Program PRESENCE using Akaike's Information Criterion. Detection probabilities for individual spotted owls ranged from 0.54 to 0.80 if barred owls were not detected during the survey season and from 0.19 to 0.71 if barred owls were detected during the survey season. Pair detection probabilities ranged from 0.27 to 0.67 if barred owls were not detected during an individual survey and from 0.09 to 0.36 if barred owls were detected during an individual survey. During the study, site occupancy probabilities for spotted owl pairs declined by approximately 50%. For all spotted owls, both singles and pairs, site occupancy probabilities declined moderately during the study. Barred owl presence was negatively associated with spotted owl detection probabilities, and it had a positive association with local-extinction probabilities for all spotted owls, both singles and pairs. Given that our study area has supported higher densities of barred owls for longer periods than other study areas, our results may provide insight into how barred owls have influenced spotted owl site occupancy dynamics in adjacent British Columbia, Canada, or will influence spotted owl site occupancy dynamics in Oregon and California, USA, in the future.     

Construction of Prediction Intervals in Location-Scale Families

Let x₁ ≤x₂ ≤x₃ … ≤x_r be the r smallest observations out of n observations from a location-scale family with density $ \frac{1}{\sigma}f\left({\frac{{x - \mu}}{\sigma}} \right) $ where μ and σ are the location and the scale parameters respectively. The goal is to construct a prediction interval of the form $ \left({\hat \mu + k_1 \hat \sigma,\,\hat \mu + k_2 \hat \sigma} \right) $ for a location-scale invariant function, T(Y) = T(Y₁, …, Y_m), of m future observations from the same distribution. Given any invariant estimators $ \hat \mu $ and $ \hat \sigma $, we have developed a general procedure for how to compute the values of k₁ and k₂. The two attractive features of the procedure are that it does not require any distributional knowledge of the joint distribution of the estimators beyond their first two raw moments and $ \hat \mu $ and $ \hat \sigma $ can be any invariant estimators of μ and σ. Examples with real data have been given and extensive simulation study showing the performance of the procedure is also offered.     

Estimation and correction of seed recovery bias from moist-soil cores

Scientists estimate seed abundances to calculate seasonal carrying capacities and assess wetland management actions for waterfowl and other wildlife using soil core samples. We evaluated recovery of known quantities of moist-soil seeds from whole and subsampled experimental core samples containing 12 seed taxa representing small, medium, and large size classes. We recovered 86.3% (SE = 1.8) of all seeds added to experimental cores; 8.3% (SE = 1.2) of seeds were destroyed during the sieving process and 5.4% (SE = 1.2) were not recovered by observers. Recovery rates varied by seed size, but not seed quantity or disproportionate ratios of seed-size classes. Overall seed recovery rates were similar between subsampled ( = 81.2%, SE = 3.6) and whole–processed core samples ( = 86.3%, SE = 1.8). We used recovery rates to generate size-specific, taxon-specific, and constant correction factors and applied each to actual core sample data. Size-specific correction factors increased seed mass estimates in the Mississippi Alluvial Valley ( = 10.1%, SE = 0.32), upper Midwest ( = 21.2%, SE = 0.61), and both regions combined ( = 15.7%, SE = 0.51) differently, as seed composition in core samples varied regionally. We suggest scientists consider using size-specific correction factors to account for seed recovery bias in core samples because these factors may be applied to a variety of taxa and produced similar mass estimates as taxon-specific correction factors. However, if data from core samples are unavailable at the resolution of seed size classes, we suggest increasing seed mass estimates by 16% to account for seed recovery bias. © 2011 The Wildlife Society.     

Bayesian Bootstrap Clones for Censored Markov Chains

In this article, we consider r observations from a non‐homogeneous censored Markov chain, with transition probability matrix P. For the product estimator of P proposed by Aalen and Johansen (1978) and Phelan (1988), we investigate the behavior of Bayesian bootstrap clones to approximate the sampling distribution of , and then construct approximate confidence interval. It is shown that the approximation based on the random‐weighted distribution is first‐order consistent. The performance of the Bayesian bootstrap clones (BBC) is also discussed by small sample simulation. Finally, we illustrate the BBC procedure in the application to the WHO malaria survey data (cf. Singer and Cohen 1970).     

Variation in spring harvest rates of male wild turkeys in New York,Ohio, and Pennsylvania

Spring harvest rates of male wild turkeys (Meleagris gallapavo) influence the number and proportion of adult males in the population and turkey population models have treated harvest as additive to other sources of mortality. Therefore, hunting regulations and their effect on spring harvest rates have direct implications for hunter satisfaction. We used tag recovery models to estimate survival rates, investigate spatial, temporal, and demographic variability in harvest rates, and assess how harvest rates may be related to management strategies and landscape characteristics. We banded 3,266 male wild turkeys throughout New York, Ohio, and Pennsylvania during 2006–2009. We found little evidence that harvest rates varied by year or management zone. The proportion of the landscape that was forested within 6.5 km of the capture location was negatively related to harvest rates; however, even though the proportion forested ranged from 0.008 to 0.96 across our study area, this corresponded to differences in harvest rates of only 2–5%. Annual survival was approximately twice as high for juveniles as adults . In turn, spring harvest rates for adult turkeys were greater for adults than juveniles . We estimated the population of male turkeys in New York and Pennsylvania ranged from 104,000 to 132,000 in all years and ranged from 63,000 to 75,000 in Ohio. Because of greater harvest rates for adult males, the proportion of adult males in the population was less than in the harvest and ranged from 0.40 to 0.81 among all states and years. The high harvest rates observed for adults may be offset by greater recruitment of juveniles into the adult age class the following year such that these states can sustain high harvest rates yet still maintain a relative high proportion of adult males in the harvest and population. © 2011 The Wildlife Society.     

Polarized fluorescence in an electric field. A model calculation with application to the geometry of the 2-hydroxy-4,4′-diamidinostilbene–DNA complex

Theoretical expressions are derived for the change in the polarized components of the fluorescence, resulting from the orientation of a rigid molecule bearing a chromophore with arbitrary angles for the absorption and transition moments \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _a $\end{document} and \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _e $\end{document} with respect to the molecular axis. The break in the symmetry relation H_V = V_H is related to the tilt angle between \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _a $\end{document} and \documentclass{article}\pagestyle{empty}\begin{document}$ \vec \mu _e $\end{document}. The theory is applied to a sonicated DNA–2-hydroxy-4,4′-diamidinostilbene complex, in the blue and red emission bands of this peculiar dye. Simultaneous measurements of linear dichroism and fluorescence lead to the determination of an angle of 47° between a fluorescent bound dye and the DNA axis, with no difference for the blue- and red-emitting species, but confirm the presence of nonfluorescent bound dye in a more perpendicular arrangement.     

Hibernal thermal ecology of eastern box turtles within a managed forest landscape

Box turtles are being extirpated from much of their former range and remaining populations often live in association with anthropogenically altered habitats. This is particularly evident at the northern distributional limit of eastern box turtles (Terrapene carolina carolina) and is an important factor to consider during the winter months when their ability to respond to microclimatic change is limited. Using temperature dataloggers, we studied the hibernal microclimate of box turtles and associated habitat following timber harvests. We monitored the body temperatures of 38 eastern box turtles and collected detailed air and soil profile temperatures of 12 box turtle hibernacula, 6 clearcuts, and 6 adjacent forested areas during the hibernal season (winter 2009–2010). We partitioned the hibernal season into 2 biologically significant thermal periods: hibernation and emergence. The mean hibernation body temperature averaged (3.28° C, SE = 0.09) and corresponded to an average depth of 10 cm. Clearcuts were consistently colder ( = 1.91° C) than forests ( = 2.68° C) and hibernacula ( = 2.77° C) during hibernation, but became the warmest areas during emergence ( = 9.96° C). We found that in the average clearcut, turtles could burrow to approximately 20 cm to attain the average hibernation body temperature or to approximately 15 cm to attain a body temperature no different than those overwintering on colder, northeast-facing slopes in the forest ( = 2.83° C). Alternatively, we found that southwest-facing slopes were warmer and if turtles chose to overwinter only in clearcuts on those slopes, they could remain shallower. All but 1 turtle overwintered in forested areas; however, our study suggests that some timber harvested areas offer various microhabitats exploitable by hibernating box turtles based on soil profile temperatures, slope aspect, and depth of hibernation. © 2012 The Wildlife Society.     

Optical anisotropy of polypeptide chains

Optical anisotropies γ² of N-t-butylacetamide (tBA), N-Methylacetamide (MA), and N, N-dimethylacetamide (DMA) have been determined from the Rayleigh ratios for depolarzed scattering by dilute solutions of the amides in p-dioxane. Traceless optical polarizability tensors \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document} for the amides are derived from these results in conjunction with the Kerr constant for tBA determined by LeGèvre and co-workers. It is shown that the tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document}_i for the glycyle unit in a polypeptide chain may be identified with \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document}MA . Methods for deriving corresponding tensors for other peptide units are indicated and the traceless polarizability tensor \documentclass{article}\pagestyle{empty}\begin{document}$ \widehat{\rm \alpha } $\end{document} for a polypeptide chain in any specified configuration is formulated.     

Survival of white-tailed deer fawns in the grasslands of the northern Great Plains

Environmental factors, such as forest characteristics, have been linked to fawn survival in eastern and southern white-tailed deer (Odocoileus virginianus) populations. In the Great Plains, less is known about how intrinsic and habitat factors influence fawn survival. During 2007–2009, we captured and radiocollared 81 fawns in north-central South Dakota and recorded 23 mortalities, of which 18 died before 1 September. Predation accounted for 52.2% of mortality; remaining mortality included human (hunting, vehicle, and farm accident; 26.1%) and hypothermia (21.7%). Coyotes (Canis latrans) accounted for 83.3% of predation on fawns. We used known-fate analysis in Program MARK to estimate summer (15 May–31 Aug) survival rates and investigated the influence of intrinsic and habitat variables on survival. We developed 2 a priori model sets, including intrinsic variables and a test of annual variation in survival (model set 1) and habitat variables (model set 2). Model set 1 indicated that summer survival varied among years (2007–2009); annual survival rates were 0.94 (SE = 0.06, n = 22), 0.78 (SE = 0.09, n = 27), and 0.54 (SE = 0.10, n = 32), respectively. Model set 2 indicated that survival was further influenced by patch density of cover habitats (Conservation Reserve Program [CRP]-grasslands, forested cover, and wetlands). Mean CRP-grassland and wetland patch density (no. patches/100 ha) were greater (P < 0.001) in home-range areas of surviving fawns ( = 1.81, SE = 0.10, n = 63; = 1.75, SE = 0.14, n = 63, respectively) than in home-range areas of fawns that died ( = 0.16, SE = 0.04, n = 18; = 1.28, SE = 0.10, n = 18, respectively). Mean forested cover patch density was less (P < 0.001) in home-range areas of surviving fawns ( = 0.77, SE = 0.10, n = 63) than in home-range areas of fawns that died ( = 1.49, SE = 0.21, n = 18). Our results indicate that management activities should focus on CRP-grassland and wetland habitats in order to maintain or improve fawn survival in the northern Great Plains, rather than forested cover composed primarily of tree plantings and shelterbelts. © 2012 The Wildlife Society.     

Equi-replicated balanced block designs generated by a balanced matrix

In this paper it is shown that if N= \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop \sum \limits_{i = 1}^{S_h} $\end{document} c_ihN_ih, where c_ih are some non-negative integer numbers and N_ih are such incidence matrices that A_h = \documentclass{article}\pagestyle{empty}\begin{document}$ \mathop \sum \limits_{i = 1}^{S_h} $\end{document} i N_ih is a balanced matrix defined by SHAH (1959), for h = 1, 2,…, p, then a block design with an incidence matrix Ñ = [N, N,…,N] is an equi-replicated balanced block design. Here the balance of a block design is defined in terms of the matrix M₀ introduced by CALI?SKI (1971).     

A stereospecific complex of poly(I) with ammonium ion

We describe conditions which lead to complete helix formation of poly(I) in the presence of NH. Binding of NH is shown to be specific in the presence of Li⁺, which does not by itself support helix formation under these conditions. The NH–poly(I) complex is characterized by uv, CD, and ir spectroscopy. The CD spectrum is strikingly different from those of the Na⁺ or K⁺ complexes, the first extremum being changed from negative for the metal ions to positive for NH. A stereospecific model is proposed for the NH–poly(I) helix in which the N of NH is located on the axis of the four-stranded helix, midway between planar tetramers formed by the bases. The model is consistent with the tetrahedral symmetry of NH, the requirement for four acceptable hydrogen bonds, the observed stability of the helix, and the accepted geometry of the backbone.     

Levels of DNA strand breaks and superoxide in phorbol ester-treated human granulocytes

Phorbol ester treatment of granulocytes triggers release of superoxide (O) and a concomitant burst of DNA strand breaks. The relationship between the amount of O and the number of DNA breaks has not previously been explored. To quantify the relatively large amount of O generated over a 40-min period by 1 × 10⁶ granulocytes/mL, a discontinuous “10-min pulse” method employing cytochrome c was used; 140 nmol O per 1 × 10⁶ cells was detected. DNA strand breaks were quantified by fluorimetric analysis of DNA unwinding (FADU). To vary the level of O released by cells, inhibitors of the respiratory burst were used. Sodium fluoride (1–10 mM) and staurosporine (2–10 nM) both inhibited O production. In both cases, however, inhibition of strand breakage was considerably more pronounced than inhibition of O. Zinc chloride (50–200 μM) inhibited both O and DNA breaks, approximately equally. Dinophysistoxin-1 (okadaic acid) inhibited O production more effectively than it inhibited DNA breaks. O dismutes to H₂O₂, a reactive oxygen species known to cause DNA breaks. The addition of catalase to remove extracellular H₂O₂ had no effect on DNA breakage. Using pulse field gel electrophoresis, few double-stranded breaks were detected compared to the number detected by FADU, indicating that about 95% of breaks were single-stranded. The level of DNA breaks is not directly related to the amount of extracellular O or H₂O₂ in PMA-stimulated granulocytes. We conclude that either an intracellular pool of these reactive oxygen species is involved in breakage or that the metabolic inhibitors are affecting a novel strand break pathway. J. Cell. Biochem. 66:219–228, 1997. © 1997 Wiley-Liss Inc.     

Regulation of changes in cytosolic Ca2+ and Na+ concentrations in rat submandibular gland acini exposed to carbachol and ATP

The relationship between cytosolic concentrations of Ca²⁺ (Ca) and Na⁺ (Na) were studied in preparations of rat submandibular and pancreatic acini loaded with the Ca²⁺-sensitive dye Fura-2 or the Na⁺-sensitive dye SBFI. Pancreatic acini showed no changes in Na during either transient or persistent changes in Ca. Increases in Ca produced by exposure of submandibular gland acini to carbachol, a muscarinic cholinergic agonist, were followed by an increase in Na after a delay of 5–10 s. When Ca²⁺ stores were mobilized without Ca²⁺ influx Na also increased, but in acini loaded with BAPTA, a nonfluorescent Ca²⁺ chelator, the transient increase in Ca²⁺ caused by mobilization of stored Ca²⁺ was virtually abolished, as was the increase in Na. In the presence of ionomycin, increases in Ca were followed by increases in Na. Ca²⁺-dependent increases in Na were abolished in Na⁺-free buffer and by the presence of furosemide, a blocker of Na⁺-K⁺-2Cl⁻ cotransport. In other studies, extracellular ATP (ATP_o) produced an increase in Ca and Na. The steady-state increase in Ca was reduced by increasing extracellular Na⁺ concentrations (Na) in dose-dependent fashion (IC₅₀ = 16.4 ± 4.7 mM Na⁺). Likewise, increasing Na reduced ATP_o-stimulated ⁴⁵Ca²⁺ uptake at steady state (IC₅₀ = 15.8 ± 9.2 mM Na⁺). Changing Na had no effect on carbachol-stimulated increases in Ca. We conclude that, in rat submandibular gland acini, ATP_o promotes an increase in Ca and Na via a common influx pathway and that, under physiologic conditions, Na⁺ significantly limits the ATP_o-stimulated increase in Ca. In the presence of carbachol, however, Na rises in Ca-dependent fashion in submandibular gland acini via stimulation of Na⁺-K⁺-2Cl⁻ cotransport. © 1996 Wiley-Liss, Inc.