Origins of Neural Progenitor Cell-Derived Axons Projecting Caudally after Spinal Cord Injury.

Neural progenitor cells (NPCs) transplanted into sites of spinal cord injury (SCI) extend large numbers of axons into the caudal host spinal cord. We determined the precise locations of neurons in the graft that extend axons into the caudal host spinal cord using AAV9-Cre-initiated retrograde tracing into floxed-TdTomato-expressing NPC grafts. 7,640 ± 630 grafted neurons extended axons to a single caudal host spinal cord site located 2 mm beyond the lesion, 5 weeks post injury. While caudally projecting axons arose from neurons located in all regions of the graft, the majority of caudally projecting graft neurons (53%) were located within the caudal one-third of the graft. Numerous host corticospinal axons formed monosynaptic projections onto caudally projecting graft neurons; however, we find that the majority of host axonal neuronal projections formed by neural progenitor cell interneuronal "relays" across sites of SCI are likely polysynaptic in nature

Development of reverse-transcription PCR techniques to analyse the density and sex ratio of gametocytes in genetically diverse Plasmodium chabaudi infections

We have developed cross-genotype and genotype-specific quantitative reverse-transcription PCR (qRT-PCR) assays to detect and quantify the number of parasites, transmission stages (gametocytes) and male gametocytes in blood stage Plasmodium chabaudi infections. Our cross-genotype assays are reliable, repeatable and generate counts that correlate strongly (R(2)s > 90%) with counts expected from blood smears. Our genotype-specific assays can distinguish and quantify different stages of genetically distinct parasite clones (genotypes) in mixed infections and are as sensitive as our cross-genotype assays. Using these assays we show that gametocyte density and gametocyte sex ratios vary during infections for two genetically distinct parasite lines (genotypes) and present the first data to reveal how sex ratio is affected when each genotype experiences competition in mixed-genotype infections. Successful infection of mosquito vectors depends on both gametocyte density and their sex ratio and we discuss the implications of competition in genetically diverse infections for transmission success

Association between smoking-related attentional bias and craving measured in the clinic and in the natural environment

Previous laboratory studies have investigated associations between attentional bias and craving, but ecological momentary assessment (EMA) may provide ecologically-valid data. This study examines whether clinic-measured attentional bias is associated with noticing smoking cues, attention to smoking, and craving assessed by EMA and whether EMA-assessed cues and attention to smoking are associated with craving in a secondary analysis of data from 100 cigarette smokers attempting cessation. Two weeks before quitting, participants completed attentional bias assessments on visual probe (VP) and Stroop tasks and completed random EMA-assessments for seven weeks thereafter. Participants completed 9,271 random assessments, averaging 3.3 prompts/day. Clinic-measured attentional bias was not associated with cues seen (VP: OR = 1.00, 95% CI = [0.99, 1.01]; Stroop: OR = 1.00, 95% CI [0.99, 1.00]), attention toward smoking (VP: OR = 1.00, 95% CI [0.99, 1.02]; Stroop: OR = 1.00, 95% CI [0.99, 1.00]), or craving (VP: OR = 1.00, 95% CI [0.99, 1.02]; Stroop: OR = 1.00, 95% CI [0.99, 1.01]). EMA responses to seeing a smoking cue (OR = 1.94, 95% CI [1.74, 2.16]) and attention toward smoking (OR = 3.69, 95% CI [3.42, 3.98]) were associated with craving. Internal reliability was higher for the Stroop (α = .75) than visual probe task (α = .20). In smokers attempting cessation, clinic measures of attentional bias do not predict noticing smoking cues, focus on smoking, or craving. However, associations exist between noticing smoking cues, attention toward smoking, and craving assessed in the moment, suggesting that attentional bias may not be a stable trait. (PsycINFO Database Recor

Can Cover Crop Use Allow Increased Levels of Corn Residue Removal for Biofuel in Irrigated and Rainfed Systems?

Corn (Zea mays L.) residue removal at high rates can result in negative impacts to soil ecosystem services. The use of cover crops could be a potential strategy to ameliorate any adverse effects of residue removal while allowing greater removal levels. Hence, the objective of this study was to determine changes in water erosion potential, soil organic C (SOC) and total N concentration, and crop yields under early- and late-terminated cover crop (CC) combined with five levels of corn residue removal after 3 years on rainfed and irrigated no-till continuous corn in Nebraska. Treatments were no CC, early- and late-terminated winter rye (Secale cereale L.) CC, and 0, 25, 50, 75, and 100% corn residue removal rates. Complete residue removal reduced mean weight diameter (MWD) of water-stable aggregates (5 cm depth) by 29% compared to no removal at the rainfed site only, suggesting increased water erosion risk at rainfed sites. Late-terminated CC significantly increased MWD of water-stable aggregates by 27 to 37% at both sites compared to no CC, but early-terminated CC had no effect. The increased MWD with late-terminated CC suggests that CC when terminated late can offset residue removal-induced risks of water erosion. Residue removal and CC did not affect SOC and total soil N concentration. Particulate organic matter increased with late-terminated CC at the irrigated site compared to no CC. Complete residue removal increased irrigated grain yield by 9% in 1 year relative to no removal. Late-terminated CC had no effect on corn yield except in 1 year when yield was 8% lower relative to no CC due to low precipitation at corn establishment. Overall, late-terminated CC ameliorates residue removal-induced increases in water erosion potential and could allow greater levels of removal without reducing corn yields in most years, in the short term, under the conditions of this study

Column classification/characterisation of strong cation exchange phases for the liquid chromatographic analysis of small molecular weight bases

A simple, rapid and robust protocol for the characterisation of strong cation exchange columns for the analysis of small molecular weight bases is described. A range of ten different phases were characterised, and the resultant selectivity and retention factors analysed using Principal Component Analysis. The score plots for the first and second principal components described 83% of the variability within the dataset. Score plots highlighted the large chromatographic differences observed between the phases, the validity of which was established using a larger range of bases. All the strong cation exchange materials demonstrated a synergistic mixed mode (i.e. ion exchange and hydrophobic) retention mechanism. Principal Component Analysis also highlighted the potential difficulty in locating suitable strong cation exchange “back-up” columns for the analysis of small molecular weight bases in that the characterised columns all displayed very different selectivities. The robustness of the protocol was confirmed by a factorial design experiment

Structural basis for inhibition of the epidermal growth factor receptor by cetuximab

SummaryRecent structural studies of epidermal growth factor receptor (EGFR) family extracellular regions have identified an unexpected mechanism for ligand-induced receptor dimerization that has important implications for activation and inhibition of these receptors. Here we describe the 2.8 Å resolution X-ray crystal structure of the antigen binding (Fab) fragment from cetuximab (Erbitux), an inhibitory anti-EGFR antibody, in complex with the soluble extracellular region of EGFR (sEGFR). The sEGFR is in the characteristic “autoinhibited” or “tethered” inactive configuration. Cetuximab interacts exclusively with domain III of sEGFR, partially occluding the ligand binding region on this domain and sterically preventing the receptor from adopting the extended conformation required for dimerization. We suggest that both these effects contribute to potent inhibition of EGFR activation

Behavior of the Escape Rate Function in Hyperbolic Dynamical Systems

For a fixed initial reference measure, we study the dependence of the escape rate on the hole for a smooth or piecewise smooth hyperbolic map. First, we prove the existence and Holder continuity of the escape rate for systems with small holes admitting Young towers. Then we consider general holes for Anosov diffeomorphisms, without size or Markovian restrictions. We prove bounds on the upper and lower escape rates using the notion of pressure on the survivor set and show that a variational principle holds under generic conditions. However, we also show that the escape rate function forms a devil's staircase with jumps along sequences of regular holes and present examples to elucidate some of the difficulties involved in formulating a general theory.Comment: 21 pages. v2 differs from v1 only by additions to the acknowledgment

Structural Evaluation of EGFR Inhibition Mechanisms for Nanobodies/VHH Domains

SummaryThe epidermal growth factor receptor (EGFR) is implicated in human cancers and is the target of several classes of therapeutic agents, including antibody-based drugs. Here, we describe X-ray crystal structures of the extracellular region of EGFR in complex with three inhibitory nanobodies, the variable domains of heavy chain only antibodies (VHH). VHH domains, the smallest natural antigen-binding modules, are readily engineered for diagnostic and therapeutic applications. All three VHH domains prevent ligand-induced EGFR activation, but use two distinct mechanisms. 7D12 sterically blocks ligand binding to EGFR in a manner similar to that of cetuximab. EgA1 and 9G8 bind an epitope near the EGFR domain II/III junction, preventing receptor conformational changes required for high-affinity ligand binding and dimerization. This epitope is accessible to the convex VHH paratope but inaccessible to the flatter paratope of monoclonal antibodies. Appreciating the modes of binding and inhibition of these VHH domains will aid in developing them for tumor imaging and/or cancer therapy

Beluga whale (Delphinapterus leucas) acoustic foraging behavior and applications for long term monitoring

This paper is not subject to U.S. copyright. The definitive version was published in Castellote, M., Mooney, A., Andrews, R., Deruiter, S., Lee, W.-J., Ferguson, M., & Wade, P. Beluga whale (Delphinapterus leucas) acoustic foraging behavior and applications for long term monitoring. Plos One, 16(11), (2021): e0260485, https://doi.org/10.1371/journal.pone.0260485.Cook Inlet, Alaska, is home to an endangered and declining population of 279 belugas (Delphinapterus leucas). Recovery efforts highlight a paucity of basic ecological knowledge, impeding the correct assessment of threats and the development of recovery actions. In particular, information on diet and foraging habitat is very limited for this population. Passive acoustic monitoring has proven to be an efficient approach to monitor beluga distribution and seasonal occurrence. Identifying acoustic foraging behavior could help address the current gap in information on diet and foraging habitat. To address this conservation challenge, eight belugas from a comparative, healthy population in Bristol Bay, Alaska, were instrumented with a multi-sensor tag (DTAG), a satellite tag, and a stomach temperature transmitter in August 2014 and May 2016. DTAG deployments provided 129.6 hours of data including foraging and social behavioral states. A total of 68 echolocation click trains ending in terminal buzzes were identified during successful prey chasing and capture, as well as during social interactions. Of these, 37 click trains were successfully processed to measure inter-click intervals (ICI) and ICI trend in their buzzing section. Terminal buzzes with short ICI (minimum ICI <8.98 ms) and consistently decreasing ICI trend (ICI increment range <1.49 ms) were exclusively associated with feeding behavior. This dual metric was applied to acoustic data from one acoustic mooring within the Cook Inlet beluga critical habitat as an example of the application of detecting feeding in long-term passive acoustic monitoring data. This approach allowed description of the relationship between beluga presence, feeding occurrence, and the timing of spawning runs by different species of anadromous fish. Results reflected a clear preference for the Susitna River delta during eulachon (Thaleichthys pacificus), Chinook (Oncorhynchus tshawytscha), pink (Oncorhynchus gorbuscha), and coho (Oncorhynchus kisutch) salmon spawning run periods, with increased feeding occurrence at the peak of the Chinook and pink salmon runs.Project funding was provided by Georgia Aquarium, the Marine Mammal Laboratory of the Alaska Fisheries Science Center (MML/AFSC). Tagging was funded by the NOAA Fisheries Office of Science and Technology’s Ocean Acoustics Program. DTAG data analysis was funded by the U.S. Marine Mammal Commission grant #16-239. Funding for collecting and analyzing Cook Inlet beluga acoustic data in Susitna Delta was provided by the National Marine Fisheries Service Section 6 Office to the Alaska Department of Fish and Game. This publication is partially funded by the Cooperative Institute for Climate, Ocean, and Ecosystem Studies (CICOES), University of Washington, under NOAA Cooperative Agreement NA15OAR4320063, Contribution No. 2021-1145