Monoclonal-based ELISA for the identification of herbicidal cyclohexanedione analogues that inhibit graminaceous acetyl coenzyme-A carboxylase

Cyclohexanediones are one of four known structural classes of herbicides that inhibit graminaceous acetyl coenzyme-A carboxylase (ACCase; EC 6.4.1.2). Five monoclonal antibodies were raised against cyclohexanediones conjugated to bovine serum albumin. Cross-reactivity studies using a homologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) against 24 cyclohexanedione analogues revealed that two monoclonal antibodies (mAb A and mAb B) could segregate the analogues into active and inactive ACCase inhibitors on the basis of the analogue concentration required to inhibit 50% of antibody binding to the coating conjugate (IC(50)). Both mAb A and mAb B were also found to cross-react with various members of the indolizidinedione structural class of ACCase inhibitors in ciELISA, suggesting that both cyclohexanediones and indolizidinediones possess features recognized by monoclonal antibodies important for the inhibition of ACCase activity. In conclusion, pharmacophore-specific antibodies may be potentially valuable screening tools for the identification of new lead chemistries in a pesticide discovery program.     

Activation of polyphosphoinositide hydrolysis in T cells by H-2 alloantigen but not MLS determinants

Murine minor lymphocyte-stimulating (Mls) determinants are cell surface antigens that stimulate strong primary T cell responses; the responding T cells display restricted T cell receptor (TCR) V beta gene usage. Interaction of T cells with mitogens or major histocompatibility complex (MHC) antigens activated the polyphosphoinositide (PI) signaling pathway, but this pathway was not triggered by Mls recognition. However, interleukin-2 (IL-2) secretion and proliferation to all three stimuli were comparable. Thus, although recognition of both allo-H-2 and Mls determinants is thought to be mediated by the TCR, these antigens appear to elicit biochemically distinct signal transduction pathways.     

Fish‐Net: Probabilistic models for fishway planning,design and monitoring to support environmentally sustainable hydropower

The construction of fishways for upstream and downstream connectivity is the preferred mitigation measure for hydropower dams and other riverine barriers. Yet empirical evidence for effective design criteria for many species is missing. We therefore assembled a group of international fishway designers and combined their knowledge with available empirical data using a formal expert elicitation protocol and Bayesian networks. The expert elicitation method we use minimizes biases typically associated with such approaches. Demonstrating our application with a case‐study on the temperate Southern Hemisphere, we use the resulting probabilistic models to predict the following, given alternative design parameters: (i) the effectiveness of technical fishways for upstream movement of migratory fish; (ii) habitat quality in nature‐like bypasses for resident fish; and (iii) rates of mortality during downstream passage of all fish through turbines and spillways. The Fish Passage Network (Fish‐Net) predicts that fishways for native species could be near 0% or near 100% efficient depending on their design, suggesting great scope for adequate mitigation. Sensitivity analyses revealed the most important parameters as follows: (i) design of attraction and entrance features of technical fishways for upstream migration; (ii) habitat preferences of resident fish in nature‐like bypasses; and (iii) susceptibility of fish to barotrauma and blade strike during turbine passage. Numerical modelling predicted that mortality rates of small‐bodied fish (50–100 mm TL) due to blade strike may be higher for Kaplan than Francis turbines. Our findings can be used to support environmentally sustainable decisions in the planning, design and monitoring stages of hydropower development.     

The 2011 magnitude 9.0 Tohoku-Oki earthquake: mosaicking the megathrust from seconds to centuries

Geophysical observations from the 2011 moment magnitude (M(w)) 9.0 Tohoku-Oki, Japan earthquake allow exploration of a rare large event along a subduction megathrust. Models for this event indicate that the distribution of coseismic fault slip exceeded 50 meters in places. Sources of high-frequency seismic waves delineate the edges of the deepest portions of coseismic slip and do not simply correlate with the locations of peak slip. Relative to the M(w) 8.8 2010 Maule, Chile earthquake, the Tohoku-Oki earthquake was deficient in high-frequency seismic radiation--a difference that we attribute to its relatively shallow depth. Estimates of total fault slip and surface secular strain accumulation on millennial time scales suggest the need to consider the potential for a future large earthquake just south of this event.     

The role of floods and droughts on riverine ecosystems under a changing climate

Floods and droughts are key driving forces shaping aquatic ecosystems. Climate change may alter key attributes of these events and consequently health and distribution of aquatic species. Improved knowledge of biological responses to different types of floods and droughts in rivers should allow the better prediction of the ecological consequences of climate change‐induced flow alterations. This review highlights that in unmodified ecosystems, the intensity and direction of biological impacts of floods and droughts vary, but the overall consequence is an increase in biological diversity and ecosystem health. To predict the impact of climate change, metrics that allow the quantitative linking of physical disturbance attributes to the directions and intensities of biological impacts are needed. The link between habitat change and the character of biological response is provided by the frequency of occurrence of the river wave characteristic—that is the event's predictability. The severity of impacts of floods is largely related to the river wave amplitude (flood magnitude), while the impact of droughts is related to river wavelength (drought duration).     

Effect of Centrifugation Technique on Post-storage Characteristics of Stallion Spermatozoa

Three ejaculates were collected from four stallions and used to compare the effects of three centrifugation methods on post-storage motility and recovery of available sperm. Two aliquots per ejaculate were diluted with skim milk-glucose (SKMG) extender to 50×10⁶ sperm/mL, placed in 50-mL conical bottom tubes, and centrifuged at either 700g for 15 minutes (700g) or 600g for 12 minutes (600g). A third aliquot was diluted 1:1 with SKMG, placed in 15-mL conical tubes, and centrifuged at 400g for 7 minutes (400g). Subsamples from each pre-treated diluted ejaculate were held at room temperature and evaluated for motility at the same time as the post-centrifugation pre-storage motility evaluation was made for treated aliquots. After centrifugation, samples from each aliquot were stored at 5°C for evaluation after 24 and 48 hours or frozen in liquid nitrogen. Percentage of available sperm harvested was higher (P ≤ .05) for aliquots centrifuged in 15-mL tubes at 400g versus 600g in 50-mL tubes. After centrifugation, total but not progressive motility of aliquots centrifuged at 700g was lower than that for noncentrifuged controls and sperm from aliquots centrifuged at 400g in 15-mL tubes. After cold storage, values for total but not progressive motility or velocity were higher (P ≤ .05) for aliquots centrifuged in 15-mL tubes at 400g compared with those centrifuged in 50-mL tubes at both 600g and 700g. Postthaw motility of frozen sperm was not different between centrifugation treatments. Poststorage percentages of intact acrosomes and detached heads did not differ because of centrifugation treatment.     

A new large scale soil DNA extraction procedure and real-time PCR assay for the detection of Sclerotium cepivorum in soil

A real-time PCR assay specific for Sclerotium cepivorum, the causal agent of white rot in onions, was developed for use with a new DNA extraction method capable of processing up to 1?kg of soil in weight. The assay was specific for S. cepivorum when tested against 24 isolates representative of 14 closely related species and other pathogens of onion. The assay was highly sensitive when used with soil DNA extracted using the new DNA extraction procedure. In three different field soils tested, a good relationship between cycle threshold (Ct) and number of sclerotia was observed (R2?=?0.89). Twenty-nine soil samples from onion and leek crops were obtained and the pathogen was detected in four samples. All four positive samples were associated with current or past outbreaks of white rot of onion. Additional assays were also used on the 29 field soil samples, Botrytis aclada and Rhizoctonia solani AG8 were also detected, in one soil sample each. Rhizoctonia solani AG2-1 was more widespread and was detected in eight different soil samples. The method is therefore suitable to quantify levels of S. cepivorum in soil samples, with the added advantage that the technique allows other soil pathogens of interest to be assayed from the same DNA sample. The bulk soil DNA extraction method described here has the potential to be used to detect soil-borne pests and pathogens for other crops in a wide range of soil types.     

Ultrasonographic and laboratory screening in clinically normal mature golden retriever dogs

Wellness and pre-anesthetic screening of blood and urine of geriatric companion animals are routinely recommended. In addition, there are occasional references to the use of imaging in clinically normal geriatric patients. However, the utility of wellness testing is not known, and there is limited information regarding the value of pre-anesthetic testing. Wellness testing, including complete blood cell count, biochemical profile, urinalysis, and abdominal ultrasound, was performed on 53 clinically normal, mature golden retriever dogs. Laboratory analysis revealed abnormalities in 54.7% (29/53) of the dogs. Abdominal ultrasound screening demonstrated abnormalities in 64.2% (34/53) of the dogs. As only a small number of dogs had follow-up diagnostic testing available, the significance of these abnormalities is unknown. Further study involving a larger cohort of animals and analysis of follow-up data is necessary to determine the utility of laboratory and imaging studies in clinically normal geriatric patients.