Fewer bowl traps and more hand netting can increase effective number of bee species and reduce excessive captures.

Reports increasingly point to substantial declines in wild bee abundance and diversity, yet there is uncertainty about how best to measure these attributes in wild bee populations. Two commonly used methods are passive trapping with bee bowls or active netting of bees on flowers, but each of these has drawbacks. Comparing the outcomes of the two methods is complicated by their uncomparable units of effort. The abundance distribution of bee species is also typically highly skewed, making it difficult to accurately assess diversity when rarer species are unlikely to be caught. The effective number of species, or Hill numbers, provides a way forward by basing the response metric on the number of equally abundant species. Our goal is to compare the effective number of bee species captured between hand netting and bowl trapping in wheatgrass prairie in South Dakota and tallgrass prairie in Minnesota, USA. Species overlap between the two methods ranged from ~40% to ~60%. Emphasis placed on rare species was important, so that 95% confidence limits overlapped between the two methods for species richness but netting exceeded trapping for Shannon's and Simpson's diversities. Netting always captured more bee species with fewer bee individuals than trapping. In most cases, the number of bees captured in bowl traps indicated substantial over-sampling, with little increase in bee species detected. Correlations between bee and floral abundance, richness, and diversity differed between netted and trapped samples. We conclude that netting and trapping together produce a more complete account of species richness, but shifting sampling emphasis from trapping to netting will result in fewer bees, but more bee species captured. Due to the different relationships between bee and floral diversities that depended on sampling method, it is unwise to compare habitat associations determined by netting with those determined by trapping.

Divergent responses of butterflies and bees to burning and grazing management in tallgrass prairies.

Butterflies and bees contribute significantly to grassland biodiversity and play important roles as pollinators and herbivores. Grassland conservation and management must be seen through the lens of insect conservation and management if these species are to thrive. In North America, grasslands are a product of climate and natural disturbances such as fire and grazing. These natural disturbances have changed considerably since European colonization and subsequent landscape fragmentation. The aim of this study was to better understand the impacts of fire and grazing management on butterfly and bee communities in tallgrass prairie, enabling land managers and conservationists to better protect and manage remnant prairie. We examined butterfly and bee abundance, species richness, and diversity in Minnesota tallgrass prairies managed by grazing or fire. In 2016 and 2017, we surveyed butterflies, bees, vegetation, and surrounding land use at 20 remnant prairies (10 burned and 10 grazed) with known management histories. Butterfly and bee abundance at our study sites were significantly negatively correlated. Butterfly abundance, but not species richness, was higher in burned than grazed prairies, and prairie-associated grass-feeding butterflies were more abundant at sites with higher plant species richness. Bee abundance was unrelated to management type but was higher at sites with sandier soils; bee species richness was positively associated with forb frequency. These findings highlight the challenges of designing management plans tailored to wide groups of pollinators and the potential pitfalls of using one group of pollinators as indicators for another. They also point to the importance of a mosaic of management practices across the prairie landscape.

Comparison of expression patterns of keratin 6, 7, 16, 17, and 19 within multiple independent isolates of As(+3)- and Cd (+2)-induced bladder cancer : keratin 6, 7, 16, 17, and 19 in bladder cancer.

This laboratory has generated a series of seven cadmium (Cd(+2))- and six arsenite (As(+3))-transformed urothelial cancer cell lines by exposure of parental UROtsa cells to each agent under similar conditions of exposure. In this study, the seven Cd(+2)-transformed cell lines were characterized for the expression of keratin 6, 16, and 17 while the six As(+3) cell lines were assessed for the expression of keratin 7 and 19. The results showed that the series of Cd(+2)-transformed cell lines and their respective transplants all had expression of keratin 6, 16, and 17 mRNA and protein. The expression of keratin 6, 16, and 17 was also correlated with areas of the urothelial tumor cells that had undergone squamous differentiation. The results also showed that four of the six As(+3)-transformed cell lines had expression of keratin 7 and 19 mRNA and protein and produced subcutaneous tumors with intense focal staining for keratin 7 and 19. The other two As(+3)-transformed cell lines had very low expression of keratin 7 mRNA and protein and produced subcutaneous tumors having no immunoreactivity for keratin 7; although keratin 19 expression was still present. The peritoneal tumors produced by one of these two cell lines regained expression of keratin 7 protein. The present results, coupled with previous studies, indicate that malignant transformation of UROtsa cells by Cd(+2) or As(+3) produce similar patterns of keratin 6, 7, 16, 17, and 19 in the resulting series of cell lines and their respective tumors.

Balancing the need for seed against invasive species risks in prairie habitat restorations.

Adequate diversity and abundance of native seed for large-scale grassland restorations often require commercially produced seed from distant sources. However, as sourcing distance increases, the likelihood of inadvertent introduction of multiple novel, non-native weed species as seed contaminants also increases. We created a model to determine an "optimal maximum distance" that would maximize availability of native prairie seed from commercial sources while minimizing the risk of novel invasive weeds via contamination. The model focused on the central portion of the Level II temperate prairie ecoregion in the Midwest US. The median optimal maximum distance from which to source seed was 272 km (169 miles). In addition, we weighted the model to address potential concerns from restoration practitioners: 1. sourcing seed via a facilitated migration strategy (i.e., direct movement of species from areas south of a given restoration site to assist species' range expansion) to account for warming due to climate change; and 2. emphasizing non-native, exotic species with a federal mandate to control. Weighting the model for climate change increased the median optimal maximum distance to 398 km (247 miles), but this was not statistically different from the distance calculated without taking sourcing for climate adaptation into account. Weighting the model for federally mandated exotic species increased the median optimal maximum distance only slightly to 293 km (182 miles), so practitioners may not need to adjust their sourcing strategy, compared to the original model. This decision framework highlights some potential inadvertent consequences from species translocations and provides insight on how to balance needs for prairie seed against those risks.

Coflowering invasive plants and a congener have neutral effects on fitness components of a rare endemic plant.

Network analyses rarely include fitness components, such as germination, to tie invasive plants to population-level effects on the natives. We address this limitation in a previously studied network of flower visitors around a suite of native and invasive plants that includes an endemic plant at Badlands National Park, South Dakota, USA. Eriogonum visheri coflowers with two abundant invasive plants, Salsola tragus and Melilotus officinalis, as well as a common congener, E. pauciflorum. Network analyses had suggested strong linkages between E. visheri and S. tragus and E. pauciflorum, with a weaker link to M. officinalis. We measured visitation, pollen deposited on stigmas, achene weight and germination over three field seasons (two for germination) in four populations (two in the final season) of E. visheri and applied in situ pollen treatments to E. visheri, adding pollen from other flowers on the same plant; flowers on other E. visheri plants; S. tragus, M. officinalis, or E. pauciflorum; open pollination; or excluding pollinators. Insect visitation to E. visheri was not affected by floral abundance of any of the focal species. Most visitors were halictid bees; one of these (Lasioglossum packeri) was the only identified species to visit E. visheri all three years. Ninety-seven percent of pollen on collected E. visheri stigmas was conspecific, but 22% of flowers had >1 grain of E. pauciflorum pollen on stigmas and 7% had >1 grain of S. tragus pollen; <1% of flowers had M. officinalis pollen on stigmas. None of the pollen treatments produced significant differences in weight or germination of E. visheri achenes. We conclude that, in contrast to the results of the network analysis, neither of the invasive species poses a threat, via heterospecific pollen deposition, to pollination of the endemic E. visheri, and that its congener provides alternative pollen resources to its pollinators.

Exotic Plant Infestation Is Associated with Decreased Modularity and Increased Numbers of Connectors in Mixed-Grass Prairie Pollination Networks.

The majority of pollinating insects are generalists whose lifetimes overlap flowering periods of many potentially suitable plant species. Such generality is instrumental in allowing exotic plant species to invade pollination networks. The particulars of how existing networks change in response to an invasive plant over the course of its phenology are not well characterized, but may shed light on the probability of long-term effects on plant-pollinator interactions and the stability of network structure. Here we describe changes in network topology and modular structure of infested and non-infested networks during the flowering season of the generalist non-native flowering plant, Cirsium arvense in mixed-grass prairie at Badlands National Park, South Dakota, USA. Objectives were to compare network-level effects of infestation as they propagate over the season in infested and non-infested (with respect to C. arvense) networks. We characterized plant-pollinator networks on 5 non-infested and 7 infested 1-ha plots during 4 sample periods that collectively covered the length of C. arvense flowering period. Two other abundantly-flowering invasive plants were present during this time: Melilotus officinalis had highly variable floral abundance in both C. arvense-infested and non-infested plots and Convolvulus arvensis, which occurred almost exclusively in infested plots and peaked early in the season. Modularity, including roles of individual species, and network topology were assessed for each sample period as well as in pooled infested and non-infested networks. Differences in modularity and network metrics between infested and non-infested networks were limited to the third and fourth sample periods, during flower senescence of C. arvense and the other invasive species; generality of pollinators rose concurrently, suggesting rewiring of the network and a lag effect of earlier floral abundance. Modularity was lower and number of connectors higher in infested networks, whether they were assessed in individual sample periods or pooled into infested and non-infested networks over the entire blooming period of C. arvense. Connectors typically did not reside within the same modules as C. arvense, suggesting that effects of the other invasive plants may also influence the modularity results, and that effects of infestation extend to co-flowering native plants. We conclude that the presence of abundantly flowering invasive species is associated with greater network stability due to decreased modularity, but whether this is advantageous for the associated native plant-pollinator communities depends on the nature of perturbations they experience.

The mechanism of insulin action on islet amyloid polypeptide fiber formation.

The pathology of type II diabetes includes the presence of cytotoxic amyloid deposits in the islets of Langerhans. The main component of these deposits, islet amyloid polypeptide (IAPP), is a hormone involved in glucose metabolism and is normally co-secreted with insulin by the beta-cells of the pancreas. Here, we perform in vitro IAPP fibrillogenesis experiments in the presence and in the absence of insulin to elucidate the mechanism by which insulin acts on fiber formation. We find that insulin is an exceptionally potent inhibitor. In contrast to the vast excess of insulin over IAPP in vivo, substoichiometric amounts of insulin inhibit seeded and unseeded reactions by more than tenfold in vitro. Unusually, the magnitude of the inhibitory effect is dependent on the concentration of insulin, yet independent of the concentration of IAPP. In addition, insulin appears to bind non-specifically to fiber surfaces, giving rise to altered morphology. IAPP fiber formation in vitro requires a minimum of three steps: fiber-independent nucleation, elongation, and fiber-dependent nucleation. Furthermore, these steps are attenuated by the presence of a dispersed-phase transition. We interpret these data in the context of the phase-mediated fibrillogenesis model (PMF) and conclude through experiment and kinetic simulation that the dominant effect of insulin is to act on the elongation portion of the reaction. These results suggest that amyloid formation in type II diabetes involves either an additional agent that acts as an accelerant, or a step that segregates IAPP from insulin.

Beclin-1 expression in normal bladder and in Cd2+ and As3+ exposed and transformed human urothelial cells (UROtsa).

The expression of beclin-1 in normal human bladder and in Cd(2+) and As(3+) exposed and transformed urothelial cells (UROtsa) was examined in this study. It was shown using a combination of real-time PCR, Western analysis and immunohistochemistry that beclin-1 was expressed in the urothelial cells of the normal bladder. It was also demonstrated that the parental UROtsa cell line expressed beclin-1 mRNA and protein at levels similar to that of the in situ urothelium. The level of beclin-1 expression underwent only modest alterations when the UROtsa cells were malignantly transformed by Cd(2+) or As(3+) or when the parental cells were exposed acutely to Cd(2+) or As(3+). While there were instances of significant alterations at individual time points and within cell line-to-cell line comparisons there was no evidence of a dose-response relationship or correlations to the phenotypic properties of the cell lines. Similar results were obtained for the expression of the Atg-5, Atg-7, Atg-12 and LC3B autophagy-related proteins. The findings provide initial evidence for beclin-1 expression in normal bladder and that large alterations in the expression of beclin-1 and associated proteins do not occur when human urothelial cells are malignantly transformed with, or exposed to, either Cd(2+) or As(3+.).