Genomic recovery lags behind demographic recovery in bottlenecked populations of the Channel Island fox, Urocyon littoralis.

With continued global change, recovery of species listed under the Endangered Species Act is increasingly challenging. One rare success was the recovery and delisting of the Channel Island fox (Urocyon littoralis) after 90%-99% population declines in the 1990s. While their demographic recovery was marked, less is known about their genetic recovery. To address genetic changes, we conducted the first multi-individual and population-level direct genetic comparison of samples collected before and after the recent bottlenecks. Using whole-exome sequencing, we found that already genetically depauperate populations were further degraded by the 1990s declines and remain low, particularly on San Miguel and Santa Rosa Islands, which underwent the most severe bottlenecks. The two other islands that experienced recent bottlenecks (Santa Cruz, and Santa Catalina islands) showed mixed results based on multiple metrics of genetic diversity. Previous island fox genomics studies showed low genetic diversity before the declines and no change after the demographic recovery, thus this is the first study to show a decrease in genetic diversity over time in U. littoralis. Additionally, we found that divergence between populations consistently increased over time, complicating prospects for using inter-island translocation as a conservation tool. The Santa Catalina subspecies is now federally listed as threatened, yet other de-listed subspecies are still recovering genetic variation which may limit their ability to adapt to changing environmental conditions. This study further demonstrates that species conservation is more complex than population size and that some island fox populations are not yet 'out of the woods'.

Widespread gene flow following range expansion in Anna's Hummingbird.

Anthropogenic changes have altered the historical distributions of many North American taxa. As environments shift, ecological and evolutionary processes can combine in complex ways to either stimulate or inhibit range expansion. Here, we examined the role of evolution in a rapid range expansion whose ecological context has been well-documented, Anna's Hummingbird (Calypte anna). Previous studies have suggested that the C. anna range expansion is the result of an ecological release facilitated by human-mediated environmental changes, where access to new food sources have allowed further filling of the abiotic niche. We examined the role of gene flow and adaptation during range expansion from their native California breeding range, north into Canada and east into New Mexico and Texas, USA. Using low coverage whole genome sequencing we found high genetic diversity, low divergence, and little evidence of selection on the northern and eastern expansion fronts. Additionally, there are no clear barriers to gene flow across the native and expanded range. The lack of selective signals between core and expanded ranges could reflect (i) an absence of novel selection pressure in the expanded range (supporting the ecological release hypothesis), (ii) swamping of adaptive variation due to high gene flow, or (iii) limitations of genome scans for detecting small shifts in allele frequencies across many loci. Nevertheless, our results provide an example where strong selection is not apparent during a rapid, contemporary range shift.

Substance Use Disorder Education: A Statewide Assessment of Core Content in Nursing Schools.

Of individuals who need treatment for substance use disorder (SUD), 12.6% do not receive it. One reason for this failure to treat is an inadequately prepared health care workforce, including nurses. To establish a state-wide baseline of SUD curricular content, we collected information about topics taught and barriers to teaching important topics using an anonymous survey sent to all 55 nursing programs in Indiana. Topics deemed important were taught more frequently, such as opioid withdrawal (60.6%). Lack of expertise was more commonly reported as a barrier than lack of time (25.3% vs. 7.1%). Findings suggest that nursing students in Indiana are not taught requisite content related to SUD. We have provided pragmatic recommendations to enhance content in schools of nursing and address lack of expertise among faculty. Leaders need to actively evaluate and augment the content of their curriculums to include SUD. [Journal of Psychosocial Nursing and Mental Health Services, 61(9), 15-23.].

Phase I trial of intravesical Bacillus Calmette-Guérin combined with intravenous pembrolizumab in recurrent or persistent high-grade non-muscle-invasive bladder cancer after previous Bacillus Calmette-Guérin treatment.

OBJECTIVES: We conducted the first phase I dose-escalation trial (NCT02324582) of intravesical Bacillus Calmette-Guérin (BCG) in combination with systemic pembrolizumab in patients with high-grade non-muscle-invasive bladder cancer (HGNMIBC) who had persistent or recurrent disease after prior intravesical therapy with BCG. The primary endpoint was the safety of this combination. The secondary endpoint was clinical activity at three months following BCG treatment. METHODS: Eighteen patients were consented for the study, five of which were screen failures. Six doses of pembrolizumab were administered every 3 weeks over 16 weeks concurrently with six weekly doses of BCG beginning at week 7. Patient safety was evaluated from the time of consent through 30 days following pembrolizumab treatment. Clinical activity was determined using cystoscopy and biopsy of suspicious lesions. RESULTS: Treatment-related adverse events included one grade 4 adverse event (AEs) (adrenal insufficiency). There were nine grade 3 AEs (chest discomfort, pulmonary embolism, arthritis, wrist edema, injection site reaction, bilateral wrist pain, cardiomyopathy, hypokalemia, urinary tract infection). There were 49 grade 1 and 30 grade 2 AEs (88% of AEs). Eleven patients finished the treatment, and two patients died during the study. Of 13 patients treated, nine patients (69%) had no evidence of disease at 3 months following BCG treatment. CONCLUSIONS: We report for the first time that combining BCG and pembrolizumab in treating HGNMIBC is safe allowing complete treatment of most patients. A phase III trial has opened to test the efficacy of this combination in HGNMIBC (KEYNOTE-676).

HIV Outbreak Control With Effective Access to Care and Harm Reduction in North Carolina, 2017-2018.

Objectives. To assess and control a potential outbreak of HIV among people who inject drugs in Western North Carolina.Methods. Disease intervention specialists offered testing for hepatitis B and hepatitis C, harm reduction materials, and linkage to care to 7 linked people recently diagnosed with HIV who also injected drugs. Contacts were offered the same services and HIV testing. HIV genotype analysis was used to characterize HIV spread. We assessed testing and care outcomes by using state surveillance information.Results. Disease intervention specialists contacted 6 of 7 linked group members and received information on 177 contacts; among 96 prioritized contacts, 42 of 96 (44%) were exposed to or diagnosed with hepatitis C, 4 of 96 (4%) had hepatitis B, and 14 of 96 (15%) had HIV (2 newly diagnosed during the investigation). HIV genotype analysis suggested recent transmission to linked group members and 1 contact. Eleven of 14 with HIV were virally suppressed following the outbreak response.Conclusions. North Carolina identified and rapidly responded to an HIV outbreak among people reporting injecting drugs. Effective HIV care, the availability of syringe exchange services, and the rapid response likely contributed to controlling this outbreak.

Using Community Engagement to Map Pathways of Opioid Use and Recovery.

We created a tool for use by the community of Tippecanoe County Indiana and their Opioid Task Force to guide their collaborative efforts in addressing the opioid crisis. A community-engaged research strategy led to the creation of a map of opioid pathways based in a systems thinking approach that considers key stakeholders and community members as system components and reflects local resources, supply, and demand for services, and the social ecology unique to this community. Organizations are able to visualize the importance of their interactions with other organizations within the community as they work together to address the crisis. This approach could be replicated in other communities and used to address other public health issues.

Practices and perceptions regarding intravenous opioid infusion and cancer pain management.

BACKGROUND: In view of the recent opioid crisis, ways to promote safe and effective opioid-related practices are needed. Faster intravenous (iv) opioid infusion rates can result in increased adverse effects and risk for nonmedical opioid use. Data on best practices regarding safe iv opioid administration for cancer pain are limited. This study examined iv opioid bolus infusion practices and perceptions about opioids in cancer pain among 4 groups of inpatient oncology nurses. METHODS: An anonymous cross-sectional survey was conducted among oncology nurses working in medical, surgical, intensive care unit (ICU), and emergency department (ED) settings. An iv opioid bolus infusion speed less than 120 seconds was considered too fast. RESULTS: The participant response rate was 59% (731 of 1234). Approximately 58%, 54%, and 58% of all nurses administered morphine, hydromorphone, and fentanyl, respectively, in less than 120 seconds. The median morphine infusion speeds were 55, 60, 60, and 85 seconds for ICU, surgical, ED, and medical unit nurses, respectively (P = .0002). The odds ratios for infusing too fast were 2.04 and 2.52 for ED (P = .039) and ICU nurses (P = .003), respectively, in comparison with medical unit nurses, and they were 0.27 and 0.18 with frequent (P = .003) and very frequent use of a timing device (P = .0001), respectively, in comparison with no use. CONCLUSIONS: More than half the nurses working in the inpatient setting reported administering iv opioids too fast. ICU nurses administered opioids the fastest. Nurses who frequently used a timing device were less likely to infuse too fast. Further research is needed to standardize and improve the safe intermittent administration of iv opioids to patients with cancer.

Isolation drives increased diversification rates in freshwater amphipods.

Vicariance and dispersal events affect current biodiversity patterns in desert springs. Whether major diversification events are due to environmental changes leading to radiation or due to isolation resulting in relict species is largely unknown. We seek to understand whether the Gammarus pecos species complex underwent major diversification events due to environmental changes in the area leading either to radiation into novel habitats, or formation of relicts due to isolation. Specifically, we tested the hypothesis that Gammarus in the northern Chihuahuan Desert of New Mexico and Texas, USA are descendants of an ancient marine lineage now containing multiple undescribed species. We sequenced a nuclear (28S) and two mitochondrial (16S, COI) genes from gammarid amphipods representing 16 desert springs in the northern Chihuahuan Desert. We estimated phylogenetic relationships, divergence times, and diversification rates of the Gammarus pecos complex. Our results revealed that the region contained two evolutionarily independent lineages: a younger Freshwater Lineage that shared a most-recent-common-ancestor with an older Saline Lineage ∼66.3  MYA (95.6-42.4  MYA). Each spring system generally formed a monophyletic clade based on the concatenated dataset. Freshwater Lineage diversification rates were 2.0-9.8 times higher than rates of the Saline Lineage. A series of post-Cretaceous colonizations by ancestral Gammarus taxa was likely followed by isolation. Paleo-geological, hydrological, and climatic events in the Neogene-to-Quaternary periods (23.03  MYA - present) in western North America promoted allopatric speciation of both lineages. We suggest that Saline Lineage populations include two undescribed Gammarus species, while the Freshwater Lineage shows repetition of fine-scale genetic structure in all major clades suggesting incipient speciation. Such ongoing speciation suggests that this region will continue to be a biodiversity hotspot for amphipods and other freshwater taxa.

Public-private differences in short-term neonatal outcomes following birth by prelabour caesarean section at early and full term.

BACKGROUND: Prelabour caesarean section (CS) at early term (370 -386  weeks) is associated with higher rates of adverse short-term neonatal outcomes and higher costs than those performed at full term (390 -406  weeks). Prelabour CS is more common in private than in public hospitals in Australia, particularly at early term. AIMS: To evaluate the impact of hospital sector (public or private) and timing of delivery on short-term neonatal outcomes following prelabour CS at term. MATERIALS AND METHODS: A retrospective cohort study of 22 954 viable singleton prelabour CS births at term (370 -406  weeks) at a single centre encompassing co-located public and private hospitals during 1998-2013 was undertaken. Propensity score analysis was used to adjust for confounding differences between sectors. The primary outcome was Neonatal Critical Care Unit (NCCU) admission with serious morbidity. Secondary outcomes included respiratory distress, vigorous resuscitation and jaundice. RESULTS: The private hospital performed prelabour CS at over double the rate of the public hospital (33.7% of all private births vs 14.7% public) and more private than public prelabour CSs occurred at early term (66.8% vs 47.9%). Public babies were more than twice as likely as private babies to require admission to NCCU with serious morbidity (adjusted odds ratio (AOR) 2.54, 95% CI 1.77-3.65) but were less likely to need vigorous resuscitation (AOR 0.53, 95% CI 0.45-0.62). Disparities in outcomes between public and private cohorts were accentuated at full term. CONCLUSION: Despite early-term prelabour CSs occurring more often in the private hospital, public babies had more adverse outcomes and treatment escalations.

A review of Yellow Dirt: A Poisoned Land and the Betrayal of the Navajos.

Yellow Dirt is a thorough account of the past and present state of the Navajo Nation with regards to uranium mining. Through a journalistic approach Judy Pasternak weaves the story of the betrayal of the Navajo people. This book highlights the impact of environment on health and this review calls all nurses to be aware of these impacts and incorporate this type of knowledge into their practice.

Fanconi anemia proteins FANCD2 and FANCI exhibit different DNA damage responses during S-phase.

Fanconi anemia (FA) pathway members, FANCD2 and FANCI, contribute to the repair of replication-stalling DNA lesions. FA pathway activation relies on phosphorylation of FANCI by the ataxia telangiectasia and Rad3-related (ATR) kinase, followed by monoubiquitination of FANCD2 and FANCI by the FA core complex. FANCD2 and FANCI are thought to form a functional heterodimer during DNA repair, but it is unclear how dimer formation is regulated or what the functions of the FANCD2-FANCI complex versus the monomeric proteins are. We show that the FANCD2-FANCI complex forms independently of ATR and FA core complex, and represents the inactive form of both proteins. DNA damage-induced FA pathway activation triggers dissociation of FANCD2 from FANCI. Dissociation coincides with FANCD2 monoubiquitination, which significantly precedes monoubiquitination of FANCI; moreover, monoubiquitination responses of FANCD2 and FANCI exhibit distinct DNA substrate specificities. A phosphodead FANCI mutant fails to dissociate from FANCD2, whereas phosphomimetic FANCI cannot interact with FANCD2, indicating that FANCI phosphorylation is the molecular trigger for FANCD2-FANCI dissociation. Following dissociation, FANCD2 binds replicating chromatin prior to-and independently of-FANCI. Moreover, the concentration of chromatin-bound FANCD2 exceeds that of FANCI throughout replication. Our results suggest that FANCD2 and FANCI function separately at consecutive steps during DNA repair in S-phase.

Urinary bother, Urinalysis, and Two-Year Efficacy Follow-Up Results of Phase I Trial of Intravesical Bacillus Calmette-Guérin Combined with Intravenous Pembrolizumab in Recurrent or Persistent High-Grade Non-Muscle-Invasive Bladder Cancer after Previous Bacillus Calmette-Guérin Treatment.

OBJECTIVE: To report urinary bother, urinalysis changes, disease-free survival (DFS), and overall survival (OS) over 2 years for subjects enrolled in a phase I dose-escalation trial (NCT02324582) of intravesical Bacillus Calmette-Guérin (BCG) in combination with systemic pembrolizumab for recurrent or persistent high-grade non-muscle invasive bladder cancer (HGNMIBC). METHODS: Eighteen patients consented to the study. Five were screen failures. Clinical activity was determined using cystoscopy and cytology with a biopsy of suspicious lesions. Urinalysis and International Prostate symptom score were assessed at pre-treatment, Week 10 (during combined BCG and pembrolizumab treatment), and 3 and 6 months from treatment completion. IPSS was analyzed using a mixed-model repeated measures analysis. A Chi-square test was used to compare urinalysis results at each interval. RESULTS: The pathologic disease stage after restaging transurethral resection and before treatment was pTa in 6 (46.2%), CIS in 6 (46.2%), and pT1 in 1 (7.7%). There was no increase in reported urinary bother throughout treatment. Quality of life measurements demonstrated no change in subjective burden. On urinalysis, we did not observe significant differences at 3 months compared to baseline evaluation. At 12 months, the DFS and OS were 69.23% and 92.31%, respectively. At 24 months, the DFS and OS were 38.46% and 92.31%, respectively. CONCLUSIONS: Treatment with BCG combined with intravenous pembrolizumab is not showing increased urinary bother or adverse urinalysis changes. Two-year response data is promising and await confirmation in the phase III study (Keynote 676).

Arabidopsis ETHE1 encodes a sulfur dioxygenase that is essential for embryo and endosperm development.

Mutations in human (Homo sapiens) ETHYLMALONIC ENCEPHALOPATHY PROTEIN1 (ETHE1) result in the complex metabolic disease ethylmalonic encephalopathy, which is characterized in part by brain lesions, lactic acidemia, excretion of ethylmalonic acid, and ultimately death. ETHE1-like genes are found in a wide range of organisms; however, the biochemical and physiological role(s) of ETHE1 have not been examined outside the context of ethylmalonic encephalopathy. In this study we characterized Arabidopsis (Arabidopsis thaliana) ETHE1 and determined the effect of an ETHE1 loss-of-function mutation to investigate the role(s) of ETHE1 in plants. Arabidopsis ETHE1 is localized in the mitochondrion and exhibits sulfur dioxygenase activity. Seeds homozygous for a DNA insertion in ETHE1 exhibit alterations in endosperm development that are accompanied by a delay in embryo development followed by embryo arrest by early heart stage. Strong ETHE1 labeling was observed in the peripheral and chalazal endosperm of wild-type seeds prior to cellularization. Therefore, ETHE1 appears to play an essential role in regulating sulfide levels in seeds.

Learning in a crisis moment: a randomized controlled trial in emergency bystander intervention.

BACKGROUND: Opioid overdose is the leading cause of injury-related death in the United States. Individuals who overdose outside of clinical settings have more positive clinical outcomes when they receive naloxone, an opioid antagonist, from a bystander as an early intervention before emergency personnel arrive. However, there is a gap in knowledge about successful instantaneous learning and intervention in a real-life stressful environment. The objective of this study is to explore the efficacy of different instructional delivery methods for bystanders in a stressful environment. We aim to evaluate which methods are most effective for instantaneous learning, successful intervention, and improved clinical outcomes. METHODS: To explore instantaneous learning in a stressful environment, we conducted a quantitative randomized controlled trial to measure how accurately individuals responded to memory-based survey questions guided by different instructional methods. Students from a large university in the Midwest (n = 157) were recruited in a public space on campus and accessed the six-question survey on their mobile devices. The intervention group competed the survey immediately while the research team created a distracting environment. The control group was asked to complete the survey later in a quiet environment. RESULTS: The intervention group correctly answered 0.72 questions fewer than the control group (p = .000, CI [0.337, 1.103]). Questions Q1 and Q5 contained direct instructions with a verbal component and showed the greatest accuracy with over 90% correct for both stressful and controlled environments. CONCLUSIONS: The variability in the responses suggests that there are environmental factors as well as instructional design features which influence instantaneous learning. The findings of this study begin to address the gap in knowledge about the effects of stress on instantaneous learning and the most effective types of instruction for untrained bystanders in emergency situations.

Assessing the Impact of Indiana Public Law 194 on Curbing the Concurrent Opioid Prescribing for Indiana Medicaid Enrollees.

Background: Several US states have introduced legislation to support the legitimate medical use of opioids while limiting misuse and diversion. One concern which has been addressed through legislation is preventing individuals from seeking opioid prescriptions concurrently from multiple providers. However, the impact of this legislation on the incidence of patients receiving concurrent prescriptions remains relatively unexplored. This study examines this phenomenon based on claims data from Medicaid enrollees and the enactment of legislation in Indiana. Methods: Indiana Medicaid claims data over the period of January 2014 to December 2019 were used to determine the changes in the percentage of individuals receiving opioid prescriptions from multiple providers within a 30-day period, that is, concurrent opioid prescription (COP) individuals. Indiana Medicaid enrollees with a diagnosis of opioid use disorder (OUD) receiving opioid prescriptions, that is, the OUD-group, were identified and separated from the enrollees without a diagnosis but receiving opioid prescriptions, that is, the non-OUD group. The mean percentages of COP individuals (with or without an OUD diagnosis) within the subset of individuals that received opioid prescriptions were compared before and after the passage of Indiana Public Law 194. Results: There were 5336 who met the criteria of COP individuals, and 2050 of those were in the OUD-group. In either group, there was a significant difference in the change in percentages (slope) before and after Indiana Public Law 194 passed. In addition, there was a significant decrease in the mean percentage of COP individuals in the non-OUD group, while the difference was not significant in the OUD group. Conclusion: Our study suggests that Indiana Public Law 194 had a positive impact on curbing COP. This study is limited by the level of details available from claims data and suggests additional studies to evaluate prescription use and prescribing practices are warranted.

Trends of repeated emergency department visits among adolescents and young adults for substance use: A repeated cross-sectional study.

Emergency Department (ED) visits for substance-related concerns among young people have been increasing in recent years. Understanding the factors related to repeated ED visits (two or more ED visits per year) for substance use concerns among young people is critical to developing a more efficient mental healthcare system that does not overburden ED and that provides efficient care for substance use patients. This study examined trends of substance use-related ED visits and factors related to repeated ED visits (two or more ED visits per year, in comparison to one ED visit per year) among adolescents and young adults (aged 13 to 25 years) in the province of Ontario, Canada. Binary logistic regression models were conducted to examine associations between hospital-related factors (hospital size, urbanicity, triage level, ED wait time) and visit status (2+ vs 1 ED visit/year), controlling for patient characteristics (age/sex). A population-based, repeated cross-sectional data over a 10-year period (2008, 2013, and 2018) was used. The proportion of substance use-related repeated ED visits significantly and consistently increased in the year 2013 and 2018 compared to 2008 (2008 = 12.52%, 2013 = 19.47%, 2018 = 20.19%). Young adult, male, medium-sized hospital, urban location, wait times longer than 6 hours, and symptom severity was associated with increased numbers of repeated ED visits. Furthermore, polysubstance use, opioid use, cocaine use, and stimulant use were strongly associated with repeated ED visits compared with the use of substances such as cannabis, alcohol and sedatives. Current findings suggest that repeated ED visits for substance use concerns could be reduced by policies that reinforce evenly distributed mental health and addiction treatment services across the provinces in rural areas and small hospitals. These services should put special efforts into developing specific (e.g., withdrawal/treatment) programming for substance-related repeated ED patients. The services should target young people using multiple psychoactive substances, stimulants and cocaine.

Opioid-Related Treatment Disparities Among Medicaid Enrollees in Indiana.

Introduction: Health care disparities based on race/ethnicity and sex can be found in a variety of settings. Our aim is to determine if there are disparities in treatment provided to Indiana Medicaid enrollees who have medically documented opioid use. Study Data and Methods: We used Medicaid reimbursement claims data to extract patients who were diagnosed with opioid use disorder (OUD) or had other medical event related to opioid use between January 2018 and March 2019. We used a two-proportion Z-test to verify the difference in the proportion of treatment provided between population subgroups. The study was approved by the Purdue University Institutional Review Board (2019-118). Study Results: Over the study period, there were 52,994 Indiana Medicaid enrollees diagnosed with OUD or documentation of another opioid related event. Only 5.41% of them received at least one type of treatment service (detoxification, psychosocial, medication assisted treatment, or comprehensive). Discussion: Although Medicaid began covering treatment services for enrollees with an OUD in Indiana at the start of 2018, very few received evidence-based services. Men and White enrollees with an OUD were in general more likely to receive services compared to women and non-White enrollees.

Untrained Bystanders Administering Drone-Delivered Naloxone: An Exploratory Study.

Introduction: Opioid overdose deaths continue to climb in the United States. Administering naloxone to an overdose victim can save their life, but rapid access to naloxone remains a barrier. Delivering naloxone to a bystander using a drone has potential to increase naloxone availability but there are still many uncertainties about this mode of delivery. Can an untrained bystander to an opioid overdose successfully administer drone delivered naloxone after viewing video instructions on the drone and how long does it take? Methods: This mixed-methods observational study, conducted in a controlled outdoor environment, simulated an opioid overdose using a mannequin (overdose victim) and panicked bystander. Untrained and medically naïve participants were instructed to call for help, move the drone from the landing spot to the mannequin, and follow the instructions provided by the drone to administer naloxone. Data was collected using video recordings, interviews, and an online survey. Time stamp data was extracted from the video for 2 time points: time for removing the naloxone from the drone and time to administer the naloxone. Interviews were audio recorded and analyzed using an inductive concept analysis approach. One interview question was coded as a binary response of anxiety/no anxiety and added to the demographic data. Results: The average time to remove and administer naloxone was 62 seconds. Anxiety during the activity was reported by 59% of the participants but there was no correlation between anxiety and time. The design of our drone and instructional video can be improved. Conclusions: We have established baseline times for completing steps in administering naloxone delivered by drone. We were able to successfully induce anxiety and have a baseline measure for what percentage of untrained bystanders may experience anxiety when involved with an emergency situation. Design of instructional materials and drone construction can contribute to anxiety and successful administration of naloxone.

Tolfenpyrad displays Francisella-targeted antibiotic activity that requires an oxidative stress response regulator for sensitivity.

IMPORTANCE: Francisella species are highly pathogenic bacteria that pose a threat to global health security. These bacteria can be made resistant to antibiotics through facile methods, and we lack a safe and protective vaccine. Given their history of development as bioweapons, new treatment options must be developed to bolster public health preparedness. Here, we report that tolfenpyrad, a pesticide that is currently in use worldwide, effectively inhibits the growth of Francisella. This drug has an extensive history of use and a plethora of safety and toxicity data, making it a good candidate for development as an antibiotic. We identified mutations in Francisella novicida that confer resistance to tolfenpyrad and characterized a transcriptional regulator that is required for sensitivity to both tolfenpyrad and reactive oxygen species.