Creation of Single Vacancies in hBN with Electron Irradiation.

Understanding electron irradiation effects is vital not only for reliable transmission electron microscopy characterization, but increasingly also for the controlled manipulation of 2D materials. The displacement cross sections of monolayer hexagonal boron nitride (hBN) are measured using aberration-corrected scanning transmission electron microscopy in near ultra-high vacuum at primary beam energies between 50 and 90 keV. Damage rates below 80 keV are up to three orders of magnitude lower than previously measured at edges under poorer residual vacuum conditions, where chemical etching appears to dominate. Notably, it is possible to create single vacancies in hBN using electron irradiation, with boron almost twice as likely as nitrogen to be ejected below 80 keV. Moreover, any damage at such low energies cannot be explained by elastic knock-on, even when accounting for the vibrations of the atoms. A theoretical description is developed to account for the lowering of the displacement threshold due to valence ionization resulting from inelastic scattering of probe electrons, modeled using charge-constrained density functional theory molecular dynamics. Although significant reductions are found depending on the constrained charge, quantitative predictions for realistic ionization states are currently not possible. Nonetheless, there is potential for defect-engineering of hBN at the level of single vacancies using electron irradiation.

Response of riparian vegetation to short- and long-term hydrologic variation.

Increasing demand for river water now conflicts with an increasing desire to maintain riparian ecosystems. Efficiently managing river flows for riparian vegetation requires an understanding of the time scale of flow effects, but this information is limited by the absence of long-term studies of vegetation change in response to flow variation. To investigate the influence of short- and long-term flow variability and dam operation on riparian vegetation, we determined the occurrence of 107 plant species in 133 permanent plots of known inundating discharge along the Gunnison River in Colorado on five different occasions between 1990 and 2013. Individual species moved up and down the gradient of inundating discharge coincident with increases and decreases in mean annual flow, and the correlations between flow and species occurrence were strongest when flows were weighted by time before vegetation sampling with a median half-life of 1.5 years. Some tall, rhizomatous, perennial species, however, responded to flows on a longer time scale. Logistic regression of species occurrence showed a significant relation with inundation duration for 70 out of 107 species. Plot species richness and total vegetative cover decreased in association with desiccation at low inundation durations and with fluvial disturbance at high inundation durations. Within-plot similarity in species occurrence between years decreased strongly with increasing inundation duration. Moderate inundation durations were dominated by tall, rhizomatous, perennial herbs, including invasive Phalaris arundinacea (reed canary grass). Over the 23-year study period, species richness declined, and the proportion of rhizomatous perennials increased, consistent with the hypothesis that decreases in flow peaks and increases in low flows caused by flow regulation have decreased establishment opportunities for disturbance-dependent species. In summary, annual-scale changes in vegetation were strongly influenced by flow variation, and decadal-scale changes were influenced by decreases in fluvial disturbance from upstream flow regulation beginning decades prior to the onset of this study.

Designing flows to enhance ecosystem functioning in heavily altered rivers.

More than a century of dam construction and water development in the western United States has led to extensive ecological alteration of rivers. Growing interest in improving river function is compelling practitioners to consider ecological restoration when managing dams and water extraction. We developed an Ecological Response Model (ERM) for the Cache la Poudre River, northern Colorado, USA, to illuminate effects of current and possible future water management and climate change. We used empirical data and modeled interactions among multiple ecosystem components to capture system-wide insights not possible with the unintegrated models commonly used in environmental assessments. The ERM results showed additional flow regime modification would further alter the structure and function of Poudre River aquatic and riparian ecosystems due to multiple and interacting stressors. Model predictions illustrated that specific peak flow magnitudes in spring and early summer are critical for substrate mobilization, dynamic channel morphology, and overbank flows, with strong subsequent effects on instream and riparian biota that varied seasonally and spatially, allowing exploration of nuanced management scenarios. Instream biological indicators benefitted from higher and more stable base flows and high peak flows, but stable base flows with low peak flows were only half as effective to increase indicators. Improving base flows while reducing peak flows, as currently proposed for the Cache la Poudre River, would further reduce ecosystem function. Modeling showed that even presently depleted annual flow volumes can achieve substantially different ecological outcomes in designed flow scenarios, while still supporting social demands. Model predictions demonstrated that implementing designed flows in a natural pattern, with attention to base and peak flows, may be needed to preserve or improve ecosystem function of the Poudre River. Improved regulatory policies would include preservation of ecosystem-level, flow-related processes and adaptive management when water development projects are considered.

Scanning transmission electron microscopy under controlled low-pressure atmospheres.

Transmission electron microscopy (TEM) is carried out in vacuum to minimize the interaction of the imaging electrons with gas molecules while passing through the microscope column. Nevertheless, in typical devices, the pressure remains at 10-7 mbar or above, providing a large number of gas molecules for the electron beam to crack, which can lead to structural changes in the sample. Here, we describe experiments carried out in a modified scanning TEM (STEM) instrument, based on the Nion UltraSTEM 100. In this instrument, the base pressure at the sample is around 2×10-10 mbar, and can be varied up to 10-6 mbar through introduction of gases directly into the objective area while maintaining atomic resolution imaging conditions. We show that air leaked into the microscope column during the experiment is efficient in cleaning graphene samples from contamination, but ineffective in damaging the pristine lattice. Our experiments also show that exposure to O2 and H2O lead to a similar result, oxygen providing an etching effect nearly twice as efficient as water, presumably due to the two O atoms per molecule. H2 and N2 environments have no influence on etching. These results show that the residual gas environment in typical TEM instruments can have a large influence on the observations, and show that chemical etching of carbon-based structures can be effectively carried out with oxygen.

Aptamer lateral flow assays for rapid and sensitive detection of cholera toxin.

Aptamers are envisioned to serve as powerful synthetic substitutes to antibodies in a variety of bioanalytical assay formats. However, lateral flow assays (LFAs) remain dominated by antibody-based strategies. In this study, a LFA for the detection of cholera toxin as a model analyte is developed and optimized using a synthetic aptamer and a naturally occurring receptor as biorecognition elements and directly compared with solely aptamer and aptamer and antibody-based alternative approaches. The aptamer (CT916) recently selected by our group, GM1 receptors and an anti-cholera toxin antibody were evaluated. Relying solely on molecules that can easily be synthesized while aiming for high sensitivity, we applied a novel combination of capture aptamer and GM1 cell receptor-labeled liposomes for cholera toxin detection, achieving a limit of detection (LOD) of 2 ng ml-1 (3σ)/10 ng ml-1 (visual) in ∼15 min. To put our novel aptasensor into perspective, we developed a competitive lateral flow assay, exploiting the competition of cholera toxin in solution with immobilized cholera toxin for binding of aptamer-coated gold nanoparticles (AuNPs) (LOD = 51 ng ml-1 (3σ)/100 ng ml-1 (visual), assay time ∼10 min). As dual simultaneously binding aptamers were not available, we designed aptamer antibody pair-based lateral flow assays using aptamer-coated AuNPs which yielded a LOD of 5 ng ml-1 (by the 3σ rule)/10 ng ml-1 (visual) in a 10 min assay and an even better LOD of 0.6 ng ml-1 (3σ)/1 ng ml-1 (visual), with an ∼20 min total assay time. All set-ups are highly specific and provide an excellent alternative for cholera toxin detection in places where professional knowledge and sophisticated equipment are not readily available and cost efficient, simple, and rapid tests are needed, while the combination of GM1 cell receptor-labeled liposomes and aptamers is clearly the most promising.

Synthesis and thermal stability of ZrO2@SiO2 core-shell submicron particles.

ZrO2@SiO2 core-shell submicron particles are promising candidates for the development of advanced optical materials. Here, submicron zirconia particles were synthesized using a modified sol-gel method and pre-calcined at 400 °C. Silica shells were grown on these particles (average size: ∼270 nm) with well-defined thicknesses (26 to 61 nm) using a seeded-growth Stöber approach. To study the thermal stability of bare ZrO2 cores and ZrO2@SiO2 core-shell particles they were calcined at 450 to 1200 °C. After heat treatments, the particles were characterized by SEM, TEM, STEM, cross-sectional EDX mapping, and XRD. The non-encapsulated, bare ZrO2 particles predominantly transitioned to the tetragonal phase after pre-calcination at 400 °C. Increasing the temperature to 600 °C transformed them to monoclinic. Finally, grain coarsening destroyed the spheroidal particle shape after heating to 800 °C. In striking contrast, SiO2-encapsulation significantly inhibited grain growth and the t → m transition progressed considerably only after heating to 1000 °C, whereupon the particle shape, with a smooth silica shell, remained stable. Particle disintegration was observed after heating to 1200 °C. Thus, ZrO2@SiO2 core-shell particles are suited for high-temperature applications up to ∼1000 °C. Different mechanisms are considered to explain the markedly enhanced stability of ZrO2@SiO2 core-shell particles.

Treatment outcomes and prognostic factors of feline splenic mast cell tumors: A multi-institutional retrospective study of 64 cases.

BACKGROUND: Mast cell tumors (MCT) are common splenic tumors in cats, but there is limited information on treatment outcomes of cats with this disease. MATERIALS AND METHODS: This retrospective study evaluated treatment outcomes in 64 cats with splenic MCT. Cats were categorized into the following treatment groups: splenectomy (A, n = 20); splenectomy with chemotherapy (B, n = 20); chemotherapy alone (C, n = 15); or supportive care (D, n = 9). RESULTS: Median tumor specific survival (MTSS) was: 856, 853, 244, 365 days for groups A, B, C, and D, respectively. The MTSS was not significantly different between the 4 groups. However, comparing cats that had splenectomy (A and B) versus those that did not (C and D), the MTSS was 856 and 342 days, respectively (p=0.008). None of the prognostic factors analyzed significantly influenced survival. CONCLUSION: Splenectomy (+/- chemotherapy) significantly prolongs survival in cats with mast cell tumors. The role of chemotherapy remains unknown.

Corrigendum: Isotope analysis in the transmission electron microscope.

This corrects the article DOI: 10.1038/ncomms13040.

Cross-Linked Gold-Nanoparticle Membrane Resonators as Microelectromechanical Vapor Sensors.

We report a novel approach for the detection of volatile compounds employing electrostatically driven drumhead resonators as sensing elements. The resonators are based on freestanding membranes of alkanedithiol cross-linked gold nanoparticles (GNPs), which are able to sorb analytes from the gas phase. Under reduced pressure, the fundamental resonance frequency of a resonator is continuously monitored while the device is exposed to varying partial pressures of toluene, 4-methylpentan-2-one, 1-propanol, and water. The measurements reveal a strong, reversible frequency shift of up to ∼10 kHz, i.e., ∼5% of the fundamental resonance frequency, when exposing the sensor to toluene vapor with a partial pressure of ∼20 Pa. As this strong shift cannot be explained exclusively by the mass uptake in the membrane, our results suggest a significant impact of analyte sorption on the pre-stress of the freestanding GNP membrane. Thus, our findings point to the possibility of designing highly sensitive resonators, which utilize sorption induced changes in the membrane's pre-stress as primary transduction mechanism.

Isotope analysis in the transmission electron microscope.

The Ångström-sized probe of the scanning transmission electron microscope can visualize and collect spectra from single atoms. This can unambiguously resolve the chemical structure of materials, but not their isotopic composition. Here we differentiate between two isotopes of the same element by quantifying how likely the energetic imaging electrons are to eject atoms. First, we measure the displacement probability in graphene grown from either 12C or 13C and describe the process using a quantum mechanical model of lattice vibrations coupled with density functional theory simulations. We then test our spatial resolution in a mixed sample by ejecting individual atoms from nanoscale areas spanning an interface region that is far from atomically sharp, mapping the isotope concentration with a precision better than 20%. Although we use a scanning instrument, our method may be applicable to any atomic resolution transmission electron microscope and to other low-dimensional materials.

Ligand Layer Engineering To Control Stability and Interfacial Properties of Nanoparticles.

The use of mixed ligand layers including poly(ethylene glycol)-based ligands for the functionalization of nanoparticles is a very popular strategy in the context of nanomedicine. However, it is challenging to control the composition of the ligand layer and maintain high colloidal and chemical stability of the conjugates. A high level of control and stability are crucial for reproducibility, upscaling, and safe application. In this study, gold nanoparticles with well-defined mixed ligand layers of α-methoxypoly(ethylene glycol)-ω-(11-mercaptoundecanoate) (PEGMUA) and 11-mercaptoundecanoic acid (MUA) were synthesized and characterized by ATR-FTIR spectroscopy and gel electrophoresis. The colloidal and chemical stability of the conjugates was tested by dynamic light scattering (DLS), small-angle X-ray scattering (SAXS), and UV/vis spectroscopy based experiments, and their interactions with cells were analyzed by elemental analysis. We demonstrate that the alkylene spacer in PEGMUA is the key feature for the controlled synthesis of mixed layer conjugates with very high colloidal and chemical stability and that a controlled synthesis is not possible using regular PEG ligands without the alkylene spacer. With the results of our stability tests, the molecular structure of the ligands can be clearly linked to the colloidal and chemical stabilization. We expect that the underlying design principle can be generalized to improve the level of control in nanoparticle surface chemistry.

Porcine liver vascular bed in Biodur E20 corrosion casts.

BACKGROUND: Pigs are frequently used as animal models in experimental medicine. To identify processes of vascular development or regression, vascular elements must be recognised and quantified in a three-dimensional (3D) arrangement. Vascular corrosion casts enable the creation of 3D replicas of vascular trees. The aim of our study was to identify suitable casting media and optimise the protocol for porcine liver vascular corrosion casting. MATERIALS AND METHODS: Mercox II® (Ladd Research, Williston, Vermont, USA) and Biodur E20® Plus (Biodur Products, Heidelberg, Germany) were tested in 4 porcine livers. The resins (volume approximately 700 mL) were injected via the portal vein. Corrosion casts were examined by macro-computed tomography, micro-computed tomography and scanning electron microscopy. RESULTS: For hepatectomies, the operating protocol was optimised to avoid gas or blood clot embolisation. We present a protocol for porcine liver vascular bed casting based on corrosion specimens prepared using Biodur E20® epoxy resin. CONCLUSIONS: Only Biodur E20®Plus appeared to be suitable for high-volume vascular corrosion casting due to its optimal permeability, sufficient processing time and minimum fragility. Biodur E20® Plus is slightly elastic, radio-opaque and alcohol-resistant. These properties make this acrylic resin suitable for not only vascular research but also teaching purposes.

Evaluation of factors associated with prolonged hospital stay and outcome of febrile neutropenic patients receiving chemotherapy: 70 cases (1997-2010).

Febrile neutropenia (FN) is an important sequela in veterinary patients receiving chemotherapy. The purpose of this study was to identify factors associated with prolonged hospital stay and outcome in canine patients developing FN secondary to chemotherapy administration. Medical records of 70 dogs treated for FN at the University of Pennsylvania from 1997 to 2010 were retrospectively evaluated. The mean interval between chemotherapy and hospitalization was 7 days. Two-thirds of treated patients had lymphoma. The majority of patients (70%) received vincristine or doxorubicin prior to the development of FN. Tachycardia at admission, complicating medical issues, G-CSF use and decreasing neutrophil count after admission were associated with prolonged hospital stay. Hypotension and G-CSF use were significantly associated with death in-hospital. Mortality was 8.5%. Identification of factors associated with prolonged hospital stay and mortality in patients with FN may enable the development of risk-adapted treatment guidelines to minimize chemotherapy-associated morbidity and mortality.

Clinical, histopathological and immunohistochemical characterization of canine indolent lymphoma.

Indolent lymphoma comprises up to 29% of all canine lymphoma; however, limited information exists regarding the subtypes and biological behaviour. This retrospective study describes the clinical characteristics, histopathological and immunohistochemical features, treatment, outcome and prognostic factors for 75 dogs with indolent lymphoma. WHO histopathological classification and immunohistochemistry (IHC) for CD79a, CD3, Ki67 and P-glycoprotein (P-gp) was performed. The most common histopathological subtype was T-zone, 61.7%, (MST 33.5 months), followed by marginal zone, 25%, (MST 21.2 months), P = 0.542. The addition of IHC to preliminary histopathological classification resulted in a revised diagnosis in 20.4% of cases. The use of systemic treatment did not influence survival, P = 0.065. Dogs treated with chlorambucil and prednisone did not reach a MST, compared with a MST of 21.6 months with CHOP-based chemotherapy, P = 0.057. The overall MST of 4.4 years confirms that this is indeed an indolent disease. However, the effect of systemic treatment must be determined through prospective trials.

Elastic three-dimensional poly (ε-caprolactone) nanofibre scaffold enhances migration, proliferation and osteogenic differentiation of mesenchymal stem cells.

OBJECTIVES: We prepared 3D poly (ε-caprolactone) (PCL) nanofibre scaffolds and tested their use for seeding, proliferation, differentiation and migration of mesenchymal stem cell (MSCs). MATERIALS AND METHODS: 3D nanofibres were prepared using a special collector for common electrospinning; simultaneously, a 2D PCL nanofibre layer was prepared using a classic plain collector. Both scaffolds were seeded with MSCs and biologically tested. MSC adhesion, migration, proliferation and osteogenic differentiation were investigated. RESULTS: The 3D PCL scaffold was characterized by having better biomechanical properties, namely greater elasticity and resistance against stress and strain, thus this scaffold will be able to find broad applications in tissue engineering. Clearly, while nanofibre layers of the 2D scaffold prevented MSCs from migrating through the conformation, cells infiltrated freely through the 3D scaffold. MSC adhesion to the 3D nanofibre PCL layer was also statistically more common than to the 2D scaffold (P < 0.05), and proliferation and viability of MSCs 2 or 3 weeks post-seeding, were also greater on the 3D scaffold. In addition, the 3D PCL scaffold was also characterized by displaying enhanced MSC osteogenic differentiation. CONCLUSIONS: We draw the conclusion that all positive effects observed using the 3D PCL nanofibre scaffold are related to the larger fibre surface area available to the cells. Thus, the proposed 3D structure of the nanofibre layer will find a wide array of applications in tissue engineering and regenerative medicine.

Prospective clinical trial to compare vincristine and vinblastine in a COP-based protocol for lymphoma in cats.

BACKGROUND: Current standard chemotherapy protocols for lymphoma in cats carry risks of gastrointestinal toxicity, which can decrease quality of life and complicate response assessment. Protocols with less gastrointestinal toxicity may improve treatment tolerance. HYPOTHESIS/OBJECTIVES: The study purpose was to compare response rate, outcome, and toxicity between cats that received vincristine or vinblastine as part of combination chemotherapy for lymphoma. We hypothesized that vinblastine would have similar efficacy, but less gastrointestinal toxicity, compared with vincristine. ANIMALS: Forty client-owned cats with confirmed diagnosis of lymphoma. METHODS: Cats were randomized to 1 of 2 treatment arms and received weekly COP-based chemotherapy for 6 months or until disease progression. Response rate, progression-free survival (PFS), lymphoma-specific survival (LSS), and incidence and severity of gastrointestinal and hematologic toxicity were compared between arms. Arm cross-over occurred if specific gastrointestinal toxicity criteria were noted. RESULTS: Cats in both arms had similar response rates, PFS, and LSS (48 versus 64 days, P = .87; 139 versus 136 days, P = .96). Cats that received vincristine were significantly more likely to switch arms based on gastrointestinal toxicity than cats that received vinblastine (44.4 versus 10.5%, P = .02). Lower baseline weight was significantly negatively associated with PFS and LSS (P = .01, P = .003, respectively). Baseline anemia was significantly negatively associated with LSS (P = .04). CONCLUSIONS AND CLINICAL IMPORTANCE: Results suggest that vinblastine is a reasonable alternative to vincristine in the treatment of some cats with lymphoma. Baseline body weight remains a significant prognostic factor for cats with lymphoma.

Efficacy of doxorubicin-based chemotherapy for non-resectable canine subcutaneous haemangiosarcoma.

Eighteen dogs with measurable subcutaneous haemangiosarcoma (SQHSA) were treated with doxorubicin-based chemotherapy. Response assessment was evaluated and compared using World Health Organization (WHO), Response Evaluation Criteria in Solid Tumours (RECIST) and tumour volume criteria. The overall response rate for all dogs was 38.8% using WHO criteria, 38.8% using RECIST criteria and 44% using tumour volume criteria. One dog had a complete response. The median response duration for all dogs was 53 days (range 13-190 days). Four dogs had complete surgical excision after neoadjuvant chemotherapy. The median progression-free interval for dogs with complete surgical excision after neoadjuvant chemotherapy was significantly longer than those not having surgical excision (207 days versus 83 days, respectively) (P = 0.003). No significant difference in metastasis-free interval or survival time was found between the groups. Doxorubicin-based chemotherapy appears to be effective for non-resectable canine SQHSA, although the response duration is relatively short.

Distraction index as a risk factor for osteoarthritis associated with hip dysplasia in four large dog breeds.

OBJECTIVE: To determine if age, breed, gender, weight or distraction index (DI) influenced the risk of radiographic osteoarthritis (OA) of canine hip dysplasia (CHD) in four common dog breeds; the American bulldog, Bernese mountain dog, Newfoundland and standard poodle. MATERIALS AND METHODS: This was a cross sectional prevalence study with 4349 dogs. Canine hips were evaluated using 3 radiographic projections: the hip-extended view, the compression view and the distraction view. The hip-extended view was examined for the presence of OA. The PennHIP distraction view was utilized to calculate the DI. For all breeds, a multiple logistic regression model incorporating age, weight, gender, and DI was created. For each breed, disease-susceptibility curves grouping dogs on the basis of age were constructed. Receiver-operating characteristic (ROC) curves were developed for each breed regardless of age. RESULTS: For all breeds, DI was the most significant risk factor for the development of OA associated with CHD. Weight and age were also significant risk factors in all four breeds, but gender was not. CLINICAL SIGNIFICANCE: Results from this study support previous findings, that irrespective of breed, the probability of radiographic OA increases with hip joint laxity as measured by the DI. Breed-specific differences in this relationship, however, warrant investigation of all breeds affected by CHD to determine inherent dependency of hip OA on joint laxity. Such findings guide veterinarians in helping dog breeders to make evidence-based breeding decisions and in informing dog owners to implement preventative treatments for CHD for dogs found to be at risk.