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The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, has evolved resistance to nearly every management tactic utilized in the field. This study investigated the resistance mechanisms in a WCR strain resistant to the Bacillus thuringiensis (Bt) protein eCry3.1Ab using dsRNA to knockdown WCR midgut genes previously documented to be associated with the resistance. ATP-binding cassette transporter (ABCC4), aminopeptidase-N, cadherin, and cathepsin-B were previously found to be differentially expressed in eCry3.1Ab-resistant WCR larvae when compared to susceptible larvae after feeding on maize expressing eCry3.1Ab and its near-isoline. Here we compared the susceptibility of resistant and susceptible WCR larvae to eCry3.1Ab protein in presence or absence of dsRNA targeting the above genes using 10-day diet overlay toxicity assays. Combining ABCC4 dsRNA with eCry3.1Ab protein increased susceptibility to Bt protein in WCR-resistant larvae, but the other three genes had no such effect. Among 65 ABC transport genes identified, several were expressed differently in resistant or susceptible WCR larvae, fed on eCry3.1Ab-expressing maize versus its isoline, that may be involved in Bt resistance. Our findings provide strong evidence that ABCC4 is indirectly involved in WCR resistance to eCry3.1Ab protein by enhancing the effects of Bt-induced toxicity. [ABSTRACT FROM AUTHOR]

Artificial diets are widely used to produce insects for research and education programs. Completed diets, in which the diets are fully made from individual ingredients and ready to use, often have high water activity, making them vulnerable to degradation. Proper storage is critical to maintaining diet quality, yet the storage conditions are not well investigated. In this study, we characterized the effects of storage conditions (temperatures and storage duration) on the quality of a diet capable of rearing both specialist and generalist insect species. The completed diet, produced by both private industry and a USDA-Agricultural Research Service laboratory, was exposed to varying temperatures during a 24-hour transit over 1600 km. After transit, it was stored at 4°C for a total storage period of 28 days. In a separate experiment, the completed diet was stored immediately after diet production at five fixed temperatures (-20, 4, 22, 25, and 33°C) for up to 28 days. For both experiments, at 5 intervals after storage (1, 7, 14, 21, and 28 days), diet quality was accessed by life history parameters (survival, molting, and weight) of western corn rootworm (Diabrotica virgifera virgifera LeConte) larvae, the most serious maize pest in the United States. Our results showed that exposure to varying temperatures between -2°C and 27°C for 24 hours had no significant impact on diet quality. However, extended storage (beyond 24 hours) at any of the fixed temperatures negatively affected diet quality. Insects reared on diets stored for over 24 hours at fixed temperatures ranging from -20°C to 33°C had significant declines in performance. Among the tested temperatures, -20°C and 4°C were found to be the most effective for preserving diet quality. At these low temperatures, there were no significant changes in insect weight and survival for diets stored within 21 and 28 days, respectively, though molting was significantly reduced within 7 days of storage. These findings provide the base of information on the storage conditions for completed diets, supporting the production of healthy insects. [ABSTRACT FROM AUTHOR]

Introduction Olfactory neuroblastoma (ONB), or esthesioneuroblastoma, is a rare neuroectodermal tumor of the nasal cavity and paranasal sinuses. Most of these tumors express somatostatin receptors (SSTRs), providing a potential target for radionuclide imaging with Ga-68 DOTATATE. However, this imaging modality has not been extensively studied in ONB. Methods We conducted a retrospective chart review of 96 endoscopic endonasal skull base surgery cases for ONB performed at our institution between 2000 and 2021. Histo (H) scores were assigned to each tumor and normalized DOTATATE standardized uptake values (nSUVs) were measured as well. Results Nine patients (5 males and 4 females) with ONB were ultimately included in the study. The average age of the patients was 50 years. All ONBs had a positive SSTR2 expression (H-score > 105; mean: 180). All ONBs showed DOTATATE avidity (mean nSUV for ONB: 6.7). However, there was no correlation between H-score and nSUV, with an r2 of 0.24 (p = 0.18). Conclusion Our study shows that SSTR2 expression is found in all ONBs with associated DOTATATE avidity, which may serve as a valuable imaging modality to monitor for recurrent and metastatic disease in ONB. [ABSTRACT FROM AUTHOR]

Purpose: We sought to compare the costs of ambulatory endovascular aneurysm repair (a-EVAR) and inpatient EVAR (i-EVAR) at up to 1-year of follow-up. Materials and Methods: A retrospective cohort study of consecutive patients undergoing elective EVAR between April 2016 and December 2018 at two academic centers. Patients planned for a-EVAR were compared with i-EVAR. Costs at 30 days and 1 year were extracted. These included operating room (OR) use, bed occupancy, laboratory and imaging, emergency department (ED) visits, readmissions, and reinterventions. Baseline characteristics were compared. Multiple regression model was used to identify predictors of increased EVAR costs. Repeated measures analysis of variance (ANOVA) was used to compare cost differences at 30 days and 1 year via an intention-to-treat analysis. Bonferroni post hoc test compared between-group differences. A p value<0.05 was considered statistically significant. Results: One hundred seventy patients were included. Most underwent percutaneous EVAR (>94%) under spinal anesthesia (>84%). Ambulatory endovascular aneurysm repair was successful in 84% (84/100). Ambulatory endovascular aneurysm repair patients (76±8 years) were younger than i-EVAR (78±9 years). They also had a smaller mean aneurysm diameter (56±6 mm) compared with i-EVAR (59±6 mm). Emergency department visits, readmissions, and reinterventions were similar up to 1 year (all p=NS). Ambulatory endovascular aneurysm repair costs showed a non-statistically significant reduction in total costs at 30 days and 1 year by 27% and 21%, respectively. Patients younger than 85 years and males had a 30-day cost reduction by 34% (p=0.027) and 33% (p=0.035), respectively with a-EVAR. Conclusions: Same-day discharge is feasible and successful in selected patients. Patients younger than 85 years and males have a short-term cost benefit with EVAR done in the ambulatory setting without increased complications or reinterventions. Clinical Impact: This study shows the overall safety of ambulatory EVAR with proper patient selection. These patient had similar post-intervention complications to inpatients. Same day discharge also resulted in short-term reduction in costs in male patients and patients younger than 85 years. [ABSTRACT FROM AUTHOR]

The critical element of a microwave heating system is the resonator, which affects the effective heating of the material to be heated. It is based on the creation of an effective electric field on the material. In order to create a homogeneous and strong electric field in the entire volume of the material to be heated, it is necessary to design a resonator of the correct dimensions. In this study, a model is derived in which the required resonator dimensions for a microwave heating system with a specific resonance frequency can be calculated analytically. The design of a rectangular microwave resonator that resonates at 2.45 GHz is given purely analytically using a derived model. The extracted analytical model is validated by simulation results for two different modes. [ABSTRACT FROM AUTHOR]

The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae) is a major pest of maize in the United States Corn Belt. Recently, resistance to Bacillus thuringiensis (Bt) maize was reported in North Dakota and increased use of Bt maize hybrids could facilitate resistance evolution in other maize-producing states. In this study, susceptibility to Bt proteins was evaluated in wild D. barberi populations from 8 fields collected in 5 different states (Minnesota, Missouri, Nebraska, Iowa, and North Dakota). Field populations were compared to a susceptible D. barberi colony in seedling and diet toxicity assays conducted with 3 concentrations of Cry3Bb1 (0.4, 4.0, and 40.0 µg/cm2) and Gpp34/Tpp35Ab1 (previously called Cry34/35Ab1; 1.4, 14.0, and 140.0 µg/cm2). The 2019 population from Meeker Co., Minnesota (MN-2019), exhibited the lowest mortality to Cry3Bb1 and also had nominally lowest mortality to Gpp34/Tpp35Ab1 at the highest concentrations tested in diet toxicity assays. Percent second instar was also highest for larvae of the Minnesota population surviving Cry3Bb1. In seedling assays, MN and IA-2018 populations exhibited the highest proportion survival and dry weight to both proteins expressed in corn. No significant differences in mortality, percent second instar, and dry weight were observed at the highest concentration for both proteins among the populations collected in in 2020. Most D. barberi populations were still highly susceptible to Cry3Bb1 and Gpp34/Tpp35Ab1 proteins based on diet and seedling assays, but resistance appears to be developing in some D. barberi populations. Now that methods are available, resistance monitoring may also be needed for D. barberi in some regions. [ABSTRACT FROM AUTHOR]

Introduction: Complications following mandibular fractures occur in 9–23% of patients. Identifying those at risk is key to prevention. Previous studies highlighted smoking, age and time from injury to presentation as risk factors but rarely recorded other possible confounders. In this paper, we use a collaborative snapshot audit to document novel risk factors and confirm established risks for complications following the treatment of mandibular fractures. Methods: The audit was carried out by 122 OMFS trainees across the UK and Ireland (49 centres) over 6 months, coordinated by the Maxillofacial Surgery Trainees Research Collaborative. Variables recorded included basic demography, medical and social history, injury mechanism and type, management and 30-day outcome. Results: Nine hundred and forty-seven (947) patients with fractured mandibles were recorded. Surgical management was carried out in 76.3%. Complications at 30 days occurred 65 (9%) of those who were managed surgically. Risk factors for complications included male sex, increasing age, any medical history, increasing number of cigarettes smoked per week, increasing alcohol use per week, worse oral hygiene and increased time from injury to presentation. Discussion: We have used a large prospective snapshot audit to confirm established risk factors and identify novel risk factors. We demonstrate that time from injury to presentation is confounded by other indicators of poor health behaviour. These results are important in designing trial protocols for management of mandibular fractures and in targeting health interventions to patients at highest risk of complications. [ABSTRACT FROM AUTHOR]

Objective: Epilepsy is a chronic disorder, and approximately 25% to 30% of dogs with epilepsy are refractory to anti‐epileptic drugs. As increased activity has been shown to reduce seizure frequency in people with epilepsy, the goal of this study was to evaluate the relationship between deviation from baseline activity and seizure incidence in dogs with epilepsy. Materials and Methods: Activity and seizure data were obtained using a canine activity monitoring device and owner observed seizure logs in 53 dogs with idiopathic epilepsy receiving anti‐epileptic drugs. Each dog's activity was individually measured, and 14‐day baseline averages were calculated. Logistic regression was performed to evaluate how an observed increase in activity, ranging from 0% to 50%, above baseline activity, affects the incidence of a seizure in the following 24 hours. Results: A total of 8540 activity days and 365 seizure days were used in the final analysis with an average of 11 seizures per dog (range 0 to 30 seizures). Seizure incidence was significantly more likely when activity was 10%, 20%, or 30% above baseline activity in the 24 hours before the day of a documented seizure [95% confidence interval (1.02 to 1.60), P=0.033; 95% confidence interval (1.08 to 1.80), P=0.010; 95% confidence interval (1.13 to 2.07), P=0.005, respectively]. However, when activity levels were 40% and 50% above baseline, the effect diminished (95% confidence interval (0.74 to 1.70), P=0.532; or 95% confidence interval (0.56 to 1.66), P=0.988, respectively). Clinical Significance: Differently than in humans, this study demonstrated that a mild to moderate increase in activity resulted in a higher seizure incidence within 24 hours in dogs with epilepsy. [ABSTRACT FROM AUTHOR]

We present long‐term density trends of the Earth's upper atmosphere at altitudes between 71 and 116 km, based on atmospheric occultations of the Crab Nebula observed with X‐ray astronomy satellites, ASCA, RXTE, Suzaku, NuSTAR, and Hitomi. The combination of the five satellites provides a time period of 28 years from 1994 to 2022. To suppress seasonal and latitudinal variations, we concentrate on the data taken in autumn (49 < doy < 111) and spring (235 < doy < 297) in the northern hemisphere with latitudes of 0°–40°. With this constraint, local times are automatically limited either around noon or midnight. We obtain four sets (two seasons × two local times) of density trends at each altitude layer. We take into account variations due to a linear trend and the 11‐year solar cycle using linear regression techniques. Because we do not see significant differences among the four trends, we combine them to provide a single vertical profile of trend slopes. We find a negative density trend of roughly −5%/decade at every altitude. This is in reasonable agreement with inferences from settling rate of the upper atmosphere. In the 100–110‐km altitude, we found an exceptionally high density decline of about −12%/decade. This peak may be the first observational evidence for strong cooling due to water vapor and ozone near 110 km, which was first identified in a numerical simulation by Akmaev et al. (2006, https://doi.org/10.1016/j.jastp.2006.03.008). Further observations and numerical simulations with suitable input parameters are needed to establish this feature. Plain Language Summary: Numerical simulations have shown that, while an increase of greenhouse gases such as CO2 in the atmosphere causes heating of the troposphere (near surface), it causes cooling of the middle and upper atmosphere, which is the so‐called "greenhouse cooling." The greenhouse cooling should result in atmospheric contraction and consequently a temporal density decrease at a fixed height. However, observational evidence for the density decrease has been scarce in the mesosphere and lower thermosphere (MLT: 80–110 km), owing to difficulty in measuring the density in this region. Here, we present the first direct measurements of long‐term variations for combined N and O atom number density in the MLT, based on atmospheric occultations of the Crab Nebula observed with X‐ray astronomy satellites. The combination of five X‐ray astronomy satellites, ASCA, RXTE, Suzaku, NuSTAR, and Hitomi, allows us to explore density trends for a long period from 1994 to 2022. We take into account variations due to a temporal linear trend and the 11‐year solar cycle, using linear regression techniques. As a result, we find a negative density trend of roughly −5%/decade at every altitude, with a local minimum of −12%/decade near 105 km. This is in reasonable agreement with the state‐of‐the‐art numerical simulations. Key Points: Time series of combined O and N densities are measured in the MLT, based on atmospheric occultations of the Crab Nebula using X‐ray astronomy satellitesThe density is decreasing everywhere, with a local minimum of −12%/decade near 105 kmThe local minimum in density trends may be due to strong cooling by water vapor and ozone, as was first predicted by Akmaev et al. (2006, https://doi.org/10.1016/j.jastp.2006.03.008) [ABSTRACT FROM AUTHOR]

Insect resistance to toxins derived from Bacillus thuringiensis (Bt) is a major issue in agriculture. Resistance to Bt has been linked to the loss of toxin binding sites within the insect, changes within the gut microbiota, and midgut tissue regeneration. Histopathological documentation of intoxication and resistance to Bt is lacking for rootworms in the genus Diabrotica (Coleoptera: Chrysomelidae), a major target of Bt corn. Here, we document the morphological response of both Bt-resistant and Bt-susceptible larval western corn rootworm, Diabrotica virgifera virgifera LeConte, to intoxication with eCry3.1Ab. Gut lumen structural differences are subtle between the two colonies when feeding on non-Bt corn. However, upon ingestion of Bt-corn roots, susceptible larvae develop symptoms indicative of gut disruption by Bt, whereas resistant larvae incur milder effects. Mild disruption of the peritrophic matrix and gut lumen is accompanied by stem cell proliferation that may lead to midgut tissue regeneration. These results help contextualize the multifaceted nature of Bt-resistance in western corn rootworm for the first time from a histopathological perspective. [ABSTRACT FROM AUTHOR]

Objective: The goal of this study was to compare sleep and seizure frequency between epileptic dogs prescribed a 20% activity increase and epileptic dogs not prescribed an activity increase. Methods: Sixty‐nine dogs receiving anti‐epileptic drug therapy were enrolled in a 6‐month prospective, randomised, placebo‐controlled clinical trial with an intention‐to‐treat analysis. A canine activity monitoring device was used to measure activity levels and sleep scores. Results: Using an intention‐to‐treat analysis, the treatment group had an average of 0.381 more seizures per month (95% CI: 0.09 to 0.68) compared with the control group, although the difference in seizure days per month was not statistically significant. In a subgroup analysis of dogs whose activity increased by at least 10%, partial compliers had 0.719 more seizures per month (95% CI: 0.22 to 1.22) and 0.581 seizure days per month (95% CI: 0.001 to 1.16) compared with the control group. Sleep scores increased by 1.2% in the treatment compared with the control group (95% CI: 0.2 to 2.3%). Conclusions: Seizure frequency and sleep score increased slightly, but significantly, in dogs with idiopathic epilepsy prescribed an increase in activity, compared with a control group. [ABSTRACT FROM AUTHOR]

The development of new biopesticides to control the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is urgent due to resistance evolution to various control methods. We tested an air-dried non-live preparation of Chromobacterium species Panama (Csp_P), against multiple corn rootworm species, including Bt-resistant and -susceptible WCR strains, northern (NCR, D. barberi Smith & Lawrence), and southern corn rootworm (SCR, D. undecimpunctata howardi Barber), in diet toxicity assays. Our results documented that Csp_P was toxic to all three corn rootworms species based on lethal (LC50), effective (EC50), and molt inhibition concentration (MIC50). In general, toxicity of Csp_P was similar among all WCR strains and ~ 3-fold less toxic to NCR and SCR strains. Effective concentration (EC50) was also similar among WCR and SCR strains, and 5-7-fold higher in NCR strains. Molt inhibition (MIC50) was similar among all corn rootworm strains except NCR diapause strain that was 2.5–6-fold higher when compared to all other strains. There was no apparent cross-resistance between Csp_P and any of the currently available Bt proteins. Our results indicate that Csp_P formulation was effective at killing multiple corn rootworm strains including Bt-resistant WCR and could be developed as a potential new management tool for WCR control. [ABSTRACT FROM AUTHOR]

Objective: Our objective was to evaluate the outcomes of endovascular treatment in patients with moderate and severe claudication due to femoropopliteal disease, that is, disease of the superficial femoral and popliteal arteries. Methods: A retrospective review of all patients with moderate and severe claudication (Rutherford 2 and 3) undergoing endovascular treatment for FP disease between January 2012 and December 2017 at two university-affiliated hospitals was performed. All procedures were performed by vascular surgeons. Primary outcomes were mortality, freedom from reintervention, major adverse limb events defined as major amputations, open surgical revascularization, or progression to chronic limb-threatening ischemia (CLTI) at 30 days, 1 year, 2 years, and last follow-up. Unadjusted odds ratios were calculated to identify variables associated with adverse outcomes, and Kaplan–Meier survival curves were used to determine mortality and freedom from reintervention. Results: Eighty-five limbs in 74 patients were identified on review. Mean age was 69.6 ± 9.8 years and 74.3% were males. At a median follow-up of 49.0 ± 25.5 months, all-cause mortality rate was 8.1% (6 patients) with 16.7% being due to cardiovascular causes. Reintervention rates were 1.2%, 16.5%, and 21.2% at 30 days, 1 year, and 2 years, respectively. Major adverse limb events occurred in 3 patients and rates were 0%, 1.2%, and 2.4% at 30 days, 1 year, and 2 years, respectively. Progression to CLTI was 0%, 1.2%, and 1.2% at 30 days, 1 year, and 2 years, respectively. Claudication had improved or resolved in 55.6% (n = 34 patients), stable in 38.9% (n = 21 patients), and worse in 5.6% (n = 3 patients) Age ≥ 70 years (OR = 4.09 (1.14–14.66), p = 0.027), TASCII A lesion (OR = 4.67 (1.14–19.17), p = 0.025), and presence of 3-vessel runoff (OR = 3.70 (1.18–11.59), p = 0.022) predicted symptoms' improvement. TASCII A lesions were less likely to require reintervention (OR = 0.23 (0.06–0.86), p = 0.020). Reintervention within 1 year (OR = 11.67 (0.98–138.94), p = 0.017), reintervention with a stent (OR = 14.40 (1.19–173.67), p = 0.008) and more than one reintervention (OR = 39.00 (2.89–526.28), p < 0.001) predicted major adverse limb events. Conclusions: Careful patient selection is important when planning endovascular treatment in patients with intermittent claudication and FP disease. This could result in symptomatic improvement in more than half of the patients. Adverse outcomes such as major adverse limb events, progression to CLTI, and amputations occur at low rates. [ABSTRACT FROM AUTHOR]

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is the most serious pest of maize (Zea mays L.) in the U.S. Corn Belt and parts of Europe. Transgenic maize hybrids expressing at least one of the four currently available insecticidal toxins from Bacillus thuringiensis (Bt) Berliner, currently the most widely adopted control method in continuous maize, have faltered due to the emergence of resistance. The resistance mechanisms of WCR to Bt toxins are not fully understood. We identified metabolic profiles of susceptible and resistant WCR larvae fed on maize hybrids expressing each of three available Cry3 proteins (eCry3Ab1, mCry3A, and Cry3Bb1) targeting corn rootworms and a control non-Bt maize via an untargeted metabolomics approach. Over 580 unique metabolites found in WCR larvae were classified into different pathways (amino acids, carbohydrates, cofactors and vitamins, energy, lipid, nucleotide, peptide, and xenobiotics). By exploring shifts in WCR larval metabolome exclusively by Bt toxins, several candidate metabolites and metabolic pathways were identified in susceptible and resistant larvae that may be involved in defense against or recovery from Bt ingestion by these larvae. These findings would provide mechanistic insights into altered metabolic pathways associated with the resistance mechanisms of WCR to Bt toxins. [ABSTRACT FROM AUTHOR]

A middle Miocene, early Barstovian land mammal age vertebrate assemblage, the Eastgate local fauna (LF), is known in the basal-most part of the Monarch Mill Formation. This rich assemblage of fossil vertebrates occurs within the Middlegate Basin in Churchill County, Nevada, USA. The Eastgate LF includes the fossil remains of fish, amphibians, reptiles, birds, and over 25 families of mammals. Previous studies on the mammalian remains have revealed several endemic taxa of rodents and carnivores. Herein, we describe the ungulates, which include two taxa from two families of Artiodactyla (Ticholeptus sp., Merycoidodontidae and Barbouromeryx trigonocorneus, Palaeomerycidae) and three taxa from three families of Perissodactyla (Equidae gen. et sp. indet., Moropus merriami, Chalicotheriidae, and Teleoceras sp., Rhinocerotidae). Independent paleobotanical evidence nearly contemporaneous with the vertebrates indicates forest and shrubland paleovegetation, and suggests that the area had been uplifted to 2700-2800 m paleoaltitude. Therefore, this local fauna adds a rare glimpse of a medium-to high-altitude vertebrate community in the intermountain western interior of North America. Temporally-restricted taxa (especially the rodent Tardontia nevadans and chalicothere Moropus merriami) reinforce the early Barstovian age of the Eastgate LF and are consistent with tephrochronological dates and radiometric analyses. The presence of Barbouromeryx trigonocorneus at Eastgate is significant in that its occurrence potentially represents a temporal range extension into the early Barstovian (previously known from latest Arikareean to middle Hemingfordian), and extends the paleobiogeographic range from the Great Plains to the Great Basin. Unequivocally, the ungulates and other mammals of the Eastgate LF support the presence of a temperate forest ecosystem in the Great Basin just subsequent to the Miocene Climatic Optimum. [ABSTRACT FROM AUTHOR]

A middle Miocene, early Barstovian land mammal age vertebrate assemblage, the Eastgate local fauna (LF), is known in the basal-most part of the Monarch Mill Formation. This rich assemblage of fossil vertebrates occurs within the Middlegate Basin in Churchill County, Nevada, USA. The Eastgate LF includes the fossil remains of fish, amphibians, reptiles, birds, and over 25 families of mammals. Previous studies on the mammalian remains have revealed several endemic taxa of rodents and carnivores. Herein, we describe the ungulates, which include two taxa from two families of Artiodactyla (Ticholeptus sp., Merycoidodontidae and Barbouromeryx trigonocorneus, Palaeomerycidae) and three taxa from three families of Perissodactyla (Equidae gen. et sp. indet., Moropus merriami, Chalicotheriidae, and Teleoceras sp., Rhinocerotidae). Independent paleobotanical evidence nearly contemporaneous with the vertebrates indicates forest and shrubland paleovegetation, and suggests that the area had been uplifted to 2700-2800 m paleoaltitude. Therefore, this local fauna adds a rare glimpse of a medium-to high-altitude vertebrate community in the intermountain western interior of North America. Temporally-restricted taxa (especially the rodent Tardontia nevadans and chalicothere Moropus merriami) reinforce the early Barstovian age of the Eastgate LF and are consistent with tephrochronological dates and radiometric analyses. The presence of Barbouromeryx trigonocorneus at Eastgate is significant in that its occurrence potentially represents a temporal range extension into the early Barstovian (previously known from latest Arikareean to middle Hemingfordian), and extends the paleobiogeographic range from the Great Plains to the Great Basin. Unequivocally, the ungulates and other mammals of the Eastgate LF support the presence of a temperate forest ecosystem in the Great Basin just subsequent to the Miocene Climatic Optimum. [ABSTRACT FROM AUTHOR]

Evolution of resistance to transgenic crops producing toxins from Bacillus thuringiensis (Bt) threatens the sustainability of the technology. Examination of resistance mechanisms has largely focused on characterization of mutations in proteins serving as Bt toxin binding sites. However, insect microbial communities have the potential to provide host resistance to pesticides in a myriad of ways. Previous findings suggest the killing mechanism of Bt relies on enteric bacteria becoming pathogenic in the disrupted gut environment of the insect following Bt intoxication. Thus, here we hypothesized that resistance to Bt would alter the microbiome composition of the insect. Previous studies have manipulated the microbiome of susceptible insects and monitored their response to Bt. In our study, we characterized the associated bacterial communities of Bt‐resistant and ‐susceptible western corn rootworms, a widespread pest of maize in the United States. We found resistant insects harbor a bacterial community that is less rich and distinct from susceptible insects. After feeding on Bt‐expressing maize, susceptible insects exhibited dysbiosis of the associated bacterial community, whereas the community within resistant insects remained relatively unchanged. These results suggest resistance to Bt produces alterations in the microbiome of the western corn rootworm that may contribute to resistance. We further demonstrated that by itself, feeding on Bt toxin‐expressing seedlings caused a shift in the microbiota. This work provides a broader picture of the effect stressors have on microbiome composition, and the potential heritable changes induced as a result of intense selection. [ABSTRACT FROM AUTHOR]

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), poses a serious threat to maize (Zea mays L.) growers in the U.S. Corn Belt. Transgenic corn expressing Bacillus thuringiensis (Bt) Berliner is the major management tactic along with crop rotation. Bt crops targeting WCR populations have been widely planted throughout the Corn Belt. Rootworms have developed resistance to nearly all management strategies including Bt corn. Therefore, there is a need for new products that are not cross-resistant with the current Bt proteins. In this study, we evaluated the susceptibility of WCR strains resistant and susceptible to Cry3Bb1 to the biological insecticide Spear-T (GS-omega/kappa-Hexatoxin-Hv1a) alone and combined with Cry3Bb1 protein. The activity of Hv1a alone was similar between Cry3Bb1-resistant and susceptible strains (LC50s = 0.95 mg/cm2 and 1.50 mg/cm2, respectively), suggesting that there is no cross-resistance with Cry3Bb1 protein. Effective concentration (EC50), molt inhibition concentration (MIC50), and inhibition concentration (IC50) values of Hv1a alone were also similar between both strains, based on non-overlapping confidence intervals. Increased mortality (64%) was observed on resistant larvae exposed to Hv1a (0.6 mg/cm2) + Cry3Bb1 protein (170.8 µg/cm2) compared to 0% mortality when exposed to Cry3Bb1 alone and 34% mortality to Hv1a alone (0.3 mg/cm2). The time of larval death was not significantly different between Hv1a alone (3.79 mg/cm2) and Hv1a (0.6 mg/cm2) + Cry3Bb1 (170.8 µg/cm2). New control strategies that are not cross-resistant with current insecticides and Bt proteins are needed to better manage the WCR, and Hv1a together with Cry3Bb1 may fit this role. [ABSTRACT FROM AUTHOR]

Host plant preference of agricultural pests may shift throughout the growing season, allowing the pests to persist on wild hosts when crops are not available. Lygus Hahn (Hemiptera: Miridae) bugs are severe pests of cotton during flowering and fruiting stages, but can persist on alternative crops, or on weed species. Diversity of digestive enzymes produced by salivary glands and gut tissues play a pivotal role in an organism's ability to utilize various food sources. Polyphagous insects produce an array of enzymes that can process carbohydrates, lipids, and proteins. In this study, the digestive enzyme repertoire of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), was identified by high-throughput sequencing followed by cDNA cloning and sequencing. This study identified 87 digestive genes, including 30 polygalacturonases (PG), one β-galactosidase, three α-glucosidases, six β-glucosidases, 28 trypsin-like proteases, three serine proteases, one apyrase-like protease, one cysteine protease, 12 lipases, and two transcripts with low similarity to a xylanase A-like genes. RNA-Seq expression profiles of these digestive genes in adult tarnished plant bugs revealed that 57 and 12 genes were differentially expressed in the salivary gland and gut (≥5-fold, P ≤ 0.01), respectively. All polygalacturonase genes, most proteases, and two xylanase-like genes were differentially expressed in salivary glands, while most of the carbohydrate and lipid processing enzymes were differentially expressed in the gut. Seven of the proteases (KF208689, KF208697, KF208698, KF208699, KF208700, KF208701, and KF208702) were not detected in either the gut or salivary glands. [ABSTRACT FROM AUTHOR]

BACKGROUND: Management of the corn pest, western corn rootworm (WCR), Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), relies heavily on the planting of transgenic corn expressing toxins produced by the bacterium Bacillus thuringiensis (Bt). This has resulted in the evolution of resistance to all of the four commercially available Bt toxins targeting coleopteran insects. In this study, we evaluated the susceptibility of a Cry34/35Ab1‐resistant WCR colony in seedling and diet toxicity assays after removal from selection for six and nine generations. In addition, female fecundity, egg fertility, adult lifespan, larval development, and adult emergence were evaluated in two Cry34/35Ab1‐resistant and two susceptible WCR colonies to assess fitness costs. RESULTS: Susceptibility to Cry34/35Ab1 was restored in a colony removed from selection after six and nine generations based on diet toxicity assays and comparisons of relative survival, head capsule width, and dry weight in plant assays. Thus, pronounced fitness costs associated with resistance to Cry34/35Ab1 were documented by susceptibility being restored within six generations. In separate studies evaluating specific fitness costs, larval fitness when reared on isoline corn did not differ between resistant and susceptible colonies. However, beetles from susceptible colonies lived longer than resistant beetles which resulted in females from susceptible colonies producing significantly more eggs than resistant colonies, with no differences in egg fertility. CONCLUSIONS: The presence of a fitness cost that may contribute to the restoration of susceptibility to Bt has not been documented in other Cry3‐resistant WCR populations and could have significant impact on the deployment of resistance management practices. Published 2021. This article is a U.S. Government work and is in the public domain in the USA. [ABSTRACT FROM AUTHOR]

Objectives/hypothesis: Facial nerve monitoring (FNM) has evolved into a widely used adjunct for many surgical procedures along the course of the facial nerve. Even though majority opinion holds that FNM reduces the incidence of iatrogenic nerve injury, there are few if any studies yielding high-level evidence and no practice guidelines on which clinicians can rely. Instead, a review of the literature and medicolegal cases reveals significant variations in methodology, training, and clinical indications.Study Design: Literature review and expert opinion.Methods: Given the lack of standard references to serve as a resource for FNM, we assembled a multidisciplinary group of experts representing more than a century of combined monitoring experience to synthesize the literature and provide a rational basis to improve the quality of patient care during FNM.Results: Over the years, two models of monitoring have become well-established: 1) monitoring by the surgeon using a stand-alone device that provides auditory feedback of facial electromyography directly to the surgeon, and 2) a team, typically consisting of surgeon, technologist, and interpreting neurophysiologist. Regardless of the setting and the number of people involved, the reliability of monitoring depends on the integration of proper technical performance, accurate interpretation of responses, and their timely application to the surgical procedure. We describe critical steps in the technical set-up and provide a basis for context-appropriate interpretation and troubleshooting of recorded signals.Conclusions: We trust this initial attempt to describe best practices will serve as a basis for improving the quality of patient care while reducing inappropriate variations.Level Of Evidence: 4 Laryngoscope, 131:S1-S42, 2021. [ABSTRACT FROM AUTHOR]

Paleontological occurrences of helodermatid lizards (Helodermatidae) are relatively uncommon and scattered in the North American fossil record. A recently discovered concentration of vertebrate fossils was exposed in a karstic fissure filling in an active rock quarry in southwestern Oklahoma. Underwater screening of the ancient cave fill yielded numerous isolated and fragmentary skeletal elements of a diversity of small, and a few large, vertebrates including unexpected remains of Heloderma sp. as well as extinct large mammals (horse, peccary, Dwarf Pronghorn), indicating Pleistocene age. Radiometric dating of the locality is not possible because of the leaching, but fossils of the Prairie Vole, Microtus ochrogaster, from the locality help to constrain the age of the deposit between 1.21 Ma and 0.01 Ma. The fossils are the first of Heloderma in the Pleistocene of Oklahoma and the southern Great Plains. Based on taphonomic evidence and the relative abundance of rodent teeth, we infer that the fossils accumulated at least partly as the remains of vertebrates fed upon by Ringtails (Bassariscus; Procyonidae) and other small carnivorous mammals. The Pleistocene occurrence of Heloderma in the southern Great Plains of North American has potential implications for the paleobiogeography of the genus and begs further investigation of fossil herpetofaunas in North America. [ABSTRACT FROM AUTHOR]

Inducing humoral, cellular and mucosal immunity is likely to improve the effectiveness of HIV-1 vaccine strategies. Here, we tested a vaccine regimen in pigtail macaques using an intranasal (i.n.) recombinant Fowl Pox Virus (FPV)-gag pol env-IL-4R antagonist prime, intramuscular (i.m.) recombinant Modified Vaccinia Ankara Virus (MVA)-gag pol-IL-4R antagonist boost followed by an i.m SOSIP-gp140 boost. The viral vector—expressed IL-4R antagonist transiently inhibited IL-4/IL-13 signalling at the vaccination site. The SOSIP booster not only induced gp140-specific IgG, ADCC (antibody-dependent cellular cytotoxicity) and some neutralisation activity, but also bolstered the HIV-specific cellular and humoral responses. Specifically, superior sustained systemic and mucosal HIV Gag-specific poly-functional/cytotoxic CD4+ and CD8+ T cells were detected with the IL-4R antagonist adjuvanted strategy compared to the unadjuvanted control. In the systemic compartment elevated Granzyme K expression was linked to CD4+ T cells, whilst Granzyme B/TIA-1 to CD8+ T cells. In contrast, the cytotoxic marker expression by mucosal CD4+ and CD8+ T cells differed according to the mucosal compartment. This vector-based mucosal IL-4R antagonist/SOSIP booster strategy, which promotes cytotoxic mucosal CD4+ T cells at the first line of defence, and cytotoxic CD4+ and CD8+ T cells plus functional antibodies in the blood, may prove valuable in combating mucosal infection with HIV-1 and warrants further investigation. [ABSTRACT FROM AUTHOR]

The northern corn rootworm (NCR), Diabrotica barberi Smith & Lawrence, is an economic pest of maize in the U.S. Corn Belt. The objective of this study was to determine the baseline susceptibility of a laboratory NCR strain to Bt proteins eCry3.1Ab, mCry3A, Cry3Bb1, and Cry34/35Ab1 using seedling, single plant, and diet-toxicity assays. Plant assays were performed in greenhouse using corn hybrids expressing one of the Bt proteins and each respective near-isoline. Diet-toxicity assays, consisting of Bt proteins overlaid onto artificial diet were also conducted. In both plant assays, significantly more larvae survived Cry34/35Ab1-expressing corn compared with all other Bt-expressing corn, and larvae that survived eCry3.1Ab-expressing corn had significantly smaller head capsule widths compared with larvae that survived Cry34/35Ab1-expressing corn. In seedling assays, larvae surviving eCry3.1Ab-expressing corn also had significantly smaller head capsule widths compared with larvae that survived mCry3A-expressing corn. Additionally, larvae that survived mCry3A-expressing corn weighed significantly more than larvae surviving eCry3.1Ab- and Cry34/35Ab1-expressing corn. In single plant assays, no significant differences in larval dry weight was observed between any of the Bt-expressing corn. In diet assays, LC50s ranged from 0.14 (eCry3.1Ab) to 10.6 µg/cm2 (Cry34/35Ab1), EC50s ranged from 0.12 (Cry34/35Ab1) to 1.57 µg/cm2 (mCry3A), IC50s ranged from 0.08 (eCry3.1Ab) to 2.41 µg/cm2 (Cry34/35Ab1), and MIC50s ranged from 2.52 (eCry3.1Ab) to 14.2 µg/cm2 (mCry3A). These results establish the toxicity of four Bt proteins to a laboratory diapausing NCR strain established prior to the introduction of Bt traits and are important for monitoring resistance evolution in NCR field populations. [ABSTRACT FROM AUTHOR]

Certain loss‐of‐function mutations in the gene encoding the lysine catabolic enzyme aldehyde dehydrogenase 7A1 (ALDH7A1) cause pyridoxine‐dependent epilepsy (PDE). Missense mutations of Glu427, especially Glu427Gln, account for ~30% of the mutated alleles in PDE patients, and thus Glu427 has been referred to as a mutation hot spot of PDE. Glu427 is invariant in the ALDH superfamily and forms ionic hydrogen bonds with the nicotinamide ribose of the NAD+ cofactor. Here we report the first crystal structures of ALDH7A1 containing pathogenic mutations targeting Glu427. The mutant enzymes E427Q, Glu427Asp, and Glu427Gly were expressed in Escherichia coli and purified. The recombinant enzymes displayed negligible catalytic activity compared to the wild‐type enzyme. The crystal structures of the mutant enzymes complexed with NAD+ were determined to understand how the mutations impact NAD+ binding. In the E427Q and E427G structures, the nicotinamide mononucleotide is highly flexible and lacks a defined binding pose. In E427D, the bound NAD+ adopts a "retracted" conformation in which the nicotinamide ring is too far from the catalytic Cys residue for hydride transfer. Thus, the structures revealed a shared mechanism for loss of function: none of the variants are able to stabilise the nicotinamide of NAD+ in the pose required for catalysis. We also show that these mutations reduce the amount of active tetrameric ALDH7A1 at the concentration of NAD+ tested. Altogether, our results provide the three‐dimensional molecular structural basis of the most common pathogenic variants of PDE and implicate strong (ionic) hydrogen bonds in the aetiology of a human disease. [ABSTRACT FROM AUTHOR]

In humans, certain mutations in the gene encoding aldehyde dehydrogenase 7A1 are associated with pyridoxine‐dependent epilepsy (PDE). Understanding the impact of PDE‐causing mutations on the structure and activity of ALDH7A1 could allow for the prediction of symptom‐severity and aid the development of patient‐specific medical treatments. Herein, we investigate the biochemical and structural consequences of PDE missense mutations targeting residues in the aldehyde substrate binding site: N167S, P169S, A171V, G174V, and W175G. All but G174V could be purified for biochemical and X‐ray crystallographic analysis. W175G has a relatively mild kinetic defect, exhibiting a fivefold decrease in kcat with no change in Km. P169S and N167S have moderate defects, characterized by catalytic efficiencies of 20‐ and 100‐times lower than wild‐type, respectively. A171V has a profound functional defect, with catalytic efficiency 2000‐times lower than wild‐type. The crystal structures of the variants are the first for any PDE‐associated mutant of ALDH7A1. The structures show that missense mutations that decrease the steric bulk of the side chain tend to create a cavity in the active site. The protein responds by relaxing into the vacant space, and this structural perturbation appears to cause misalignment of the aldehyde substrate in W175G and N167S. The P169S structure is nearly identical to that of the wild‐type enzyme; however, analysis of B‐factors suggests the catalytic defect may result from altered protein dynamics. The A171V structure suggests that the potential for steric clash with Val171 prevents Glu121 from ion pairing with the amino group of the aldehyde substrate. Enzymes: Aldehyde dehydrogenase 7A1 (EC1.2.1.31). Databases: Coordinates have been deposited in the Protein Data Bank under the following accession codes: 6O4B, 6O4C, 6O4D, 6O4E, 6O4F, 6O4G, 6O4H. [ABSTRACT FROM AUTHOR]

We report on a new modular sensing approach in which complex emulsions serve as efficient transducers in optical evanescent field-based sensing devices. Specifically, we leverage the tunable refractive index upon chemically triggered changes in droplet morphology or orientation. Variations in the optical coupling result in readily detectable changes in the light transmitted from a waveguide. [ABSTRACT FROM AUTHOR]

Gopherus polyphemus populations are diminishing throughout their range due to urbanization, fragmentation, and poor habitat management. Increased population densities, poor habitat quality, and lack of fire may influence disease transmission. Parasite roles within wild tortoise populations are largely unknown; despite evidence these pathogens may pose significant health risks. This study provides a baseline of gopher tortoise intestinal parasites across South Florida and reports on how varying environmental and tortoise characteristics may affect intestinal parasite species prevalence and approximate loads. Tortoise fecal samples were taken from six tortoise populations across five South Florida sites. Seven species of intestinal parasites were discovered from 123 tortoises. Identified parasites include endohelminths such as cyathostomes, pinworms, ascarids, flukes, and protozoans including Eimeria, Cryptosporidium, and Amoeba species. Significant differences in parasite prevalence and loads were seen between sites, while parasitism among sex, size class, and habitat type remained relatively ubiquitous. [ABSTRACT FROM AUTHOR]

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is considered the most important maize (Zea mays L.) pest in the U.S. Corn Belt. Bioassays testing susceptibility to Bacillus thuringiensis Berliner (Bt) and other toxins of corn rootworm larvae often rely on artificial diet formulations. Successful bioassays on artificial diet for corn rootworm have sometimes been challenging because of microbial contamination. Toward the long-term goal of developing a universal artificial diet for western corn rootworm larvae, we compared larval survival, dry weight, and percentage of molt in 10-d bioassays from six current diets of which we were aware. In addition, as part of longer term rearing efforts, we recorded molting over an extended period of development (60 d). Six different artificial diets, including four proprietary industry diets (A, B, C, and D), the first published artificial diet for western corn rootworm (Pleau), and a new diet (WCRMO-1) were evaluated. Western corn rootworm larval survival was above 90% and contamination was 0% on all diets for 10 d. Diet D resulted in the greatest dry weight and percentage molting when compared with the other diets. Although fourth-instar western corn rootworm larvae have not been documented previously (only three instars have been previously documented), as many as 10% of the larvae from Diet B molted into a fourth instar prior to pupating. Overall, significant differences were found among artificial diets currently used to screen western corn rootworm. In order for data from differing toxins to be compared, a single, reliable and high-quality western corn rootworm artificial diet should eventually be chosen by industry, academia, and the public as a standard for bioassays. [ABSTRACT FROM AUTHOR]

Larval Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) were exposed to seven different entomopathogenic nematode species to test their potential infectivity in a laboratory setting. Known D. virgiferainfecting nematode species Heterorhabditis bacteriophora Poinar, Heterorhabditis megidis Poinar, Jackson & Klein, Steinernema feltiae Filipjev, and Steinernema carpocapsae Weiser were tested in a concerted experiment alongside Steinernema diaprepesi Nguyen & Duncan, Steinernema riobrave Cabanillas, Poinar & Raulston, and a Missouri wild-type H. bacteriophora which have not been previously tested on D. virgifera. The species S. rarum Doucet was tested separately for D. virgifera infectivity. Third-instar D. virgifera were exposed to either 60 or 120 nematodes per larva for 6 d. Following exposure, mortality was recorded and larvae were examined to determine the presence of active nematode infections. Results indicated a significantly higher proportion of larvae with active infections from the Heterorhabditidae species and S. diaprepesi than the other Steinernematidae species for both exposure rates; mortality data indicated a similar trend. Steinernema rarum showed almost no infectivity in laboratory experiments. [ABSTRACT FROM AUTHOR]

Abstract: We designed a planar monopole antenna with an impedance matching part and two rectangular parasitic elements. Parasitic elements significantly increased radiation bandwidth and gain. Measured fractional bandwidth and maximum gain for the case with parasitic elements were 129.9% and 5.67 dB, respectively. Antenna operated between 675 MHz and 3177.5 MHz. [ABSTRACT FROM AUTHOR]

It may be useful to develop prodrugs that are selectively activated by oxidative stress in cancer cells to release cell-killing reactive intermediates. However, relatively few chemical strategies exist for the activation of prodrugs under conditions of oxidative stress. Here we provide evidence for a novel process in which oxidation of a thiol residue in the natural product leinamycin E1 by H2O2 and other byproducts of cellular oxidative stress initiates generation of an episulfonium ion that selectively alkylates guanine residues in duplex DNA. [ABSTRACT FROM AUTHOR]

Manufacturing processes for biological molecules in the research laboratory have failed to keep pace with the rapid advances in automization and parellelization. We report the development of a digital-to-biological converter for fully automated, versatile and demand-based production of functional biologics starting from DNA sequence information. Specifically, DNA templates, RNA molecules, proteins and viral particles were produced in an automated fashion from digitally transmitted DNA sequences without human intervention. [ABSTRACT FROM AUTHOR]