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Objective History and physical examination findings can be unreliable for prediction of genitourinary tract infections and differentiation of urinary tract infections from sexually transmitted infections (STIs). The study objective was to develop a prediction tool to more accurately identify patients with STIs. Methods A retrospective review of 64,490 emergency department (ED) encounters between April 18, 2014, and March 7, 2017, where patients age 18 years or older had urinalysis and urine culture or testing for gonorrhea, chlamydia, or trichomonas, was used to develop a prediction model for men and women with Neisseria gonorrhoeae or Chlamydia trachomatis, or both, and for women with Trichomonas vaginalis. The data set was randomly divided into two-thirds discovery and one-third validation. Groups were assigned through a random number generator. Backward step regression modeling was used to identify the best model for each outcome. Results With use of age, race, marital status, and findings from vaginal wet preparation (white blood cells [WBCs], clue cells, and yeast) and urinalysis (squamous epithelial cells, protein, leukocyte esterase, and WBCs), the models had areas under the receiver operating characteristic curve of 0.80 for men with N gonorrhoeae or C trachomatis, or both; 0.75 for women with N gonorrhoeae or C trachomatis, or both; and 0.73 for women with T vaginalis. Conclusions The model estimated likelihood of ED patients having STIs was reasonably accurate with a limited number of demographic and laboratory variables. In the absence of point-of-care STI testing, use of a prediction tool for STIs may improve antimicrobial stewardship.

This Letter reports results from the first long-baseline search for sterile antineutrinos mixing in an accelerator-based antineutrino-dominated beam. The rate of neutral-current interactions in the two NOvA detectors, at distances of 1 km and 810 km from the beam source, is analyzed using an exposure of $12.51\times10^{20}$ protons-on-target from the NuMI beam at Fermilab running in antineutrino mode. A total of $121$ of neutral-current candidates are observed at the Far Detector, compared to a prediction of $122\pm11$(stat.)$\pm15$(syst.) assuming mixing between three active flavors. No evidence for $\bar{\nu}_{\mu}\rightarrow\bar{\nu}_{s}$ oscillation is observed. Interpreting this result within a 3+1 model, constraints are placed on the mixing angles ${\theta}_{24} < 25^{\circ}$ and ${\theta}_{34} < 32^{\circ}$ at the 90% C.L. for $0.05$eV$^{2} \leq \Delta m^{2}_{41} \leq 0.5$eV$^{2}$, the range of mass splittings that produces no significant oscillations at the Near Detector. These are the first 3+1 confidence limits set using long-baseline accelerator antineutrinos., Comment: 8 pages, 4 figures

Introduction Epididymitis and orchitis are illnesses characterized by pain and inflammation of the epididymis and testicle. They represent the most common causes of acute scrotal pain in the outpatient setting. Epididymitis and orchitis have both infectious and noninfectious causes, with most cases being secondary to the invasive pathogens chlamydia, gonorrhea, and Escherichia coli (E.coli). The study's objective was to examine the epidemiology and clinical characteristics of men diagnosed with epididymitis or orchitis in a United States emergency department. Methods We examined a dataset of 75,000 emergency department (ED) patient encounters from a single health system in Northeast Ohio who underwent nucleic acid amplification testing (NAAT) for chlamydia, gonorrhea, or trichomonas, or who received a urinalysis and urine culture. All patients were ≥18 years of age, and all encounters took place between April 18, 2014, and March 7, 2017. The analysis only included men receiving an ED diagnosis of epididymitis, orchitis, or both. We evaluated laboratory and demographic data using univariable and multivariable analyses. Results There were 1.3% (256/19,308) of men in the dataset diagnosed with epididymitis, orchitis, or both. Only 50.1% (130/256) of men diagnosed with epididymitis, orchitis, or both were tested for gonorrhea and chlamydia during their clinical encounter, and among those 13.8% (18/130) were positive. Chlamydia (12.3% [16/130]) was more common than both gonorrhea (3.1% [4/129]) and trichomonas (8.8% [3/34]) among men

Excess inflammation and canonical BMP receptor (BMPR) signaling are coinciding hallmarks of the early stages of injury-induced endochondral heterotopic ossification (EHO), especially in the rare genetic disease fibrodysplasia ossificans progressiva (FOP). Multiple inflammatory signaling pathways can synergistically enhance BMP-induced Smad1/5/8 activity in multiple cell types, suggesting the importance of pathway crosstalk in EHO and FOP. Toll-like receptors (TLRs) and IL-1 receptors mediate many of the earliest injury-induced inflammatory signals largely via MyD88-dependent pathways. Thus, the hypothesis that MyD88-dependent signaling is required for EHO was tested in vitro and in vivo using global or Pdgfrα-conditional deletion of MyD88 in FOP mice. As expected, IL-1β or LPS synergistically increased Activin A (ActA)-induced phosphorylation of Smad 1/5 in fibroadipoprogenitors (FAPs) expressing Alk2R206H. However, conditional deletion of MyD88 in Pdgfrα-positive cells of FOP mice did not significantly alter the amount of muscle injury-induced EHO. Even more surprisingly, injury-induced EHO was not significantly affected by global deletion of MyD88. These studies demonstrate that MyD88-dependent signaling is dispensable for injury-induced EHO in FOP mice.

The use of claims databases for research after total hip and knee arthroplasty (THA, TKA) has increased exponentially. These studies rely on accurate coding, and inadvertent inclusion of patients with nonroutine indications may influence results. The purpose of this study was to evaluate the complexity of THA and TKA captured by CPT code and determine if complication rates vary based on the indication.The NSQIP database was queried using CPT codes 21730 and 27447 to identify patients undergoing THA and TKA from 2018 to 2019. The surgical indication was classified based on the ICD-10 diagnosis code as routine primary, complex primary, inflammatory, fracture, oncologic, revision, infection, or indeterminant. Patient factors and 30-day complications, readmission, reoperation, and wound complications were compared.A total of 86,009 THA patients had 703 ICD-10 diagnosis codes and 91.4% were routine primary indications. Complication rates were: routine primary 7.4%, complex primary 11.3%, inflammatory 12.5%, fracture 23.9%, oncologic 32.4%, revision 26.9%, infection 38.7%, and indeterminant 10.3% (P.0001). 137,500 TKA patients had 552 ICD-10 diagnosis codes and 96.1% were routine primary cases. Complication rates were: routine primary 5.9%, complex primary 8.0%, inflammatory 7.2%, fracture 38.9%, oncologic 32.7%, revision 13.3%, infection 37.7%, and indeterminant 9.6% (P.0001). Routine primary arthroplasty had significantly lower rates of reoperation, readmission, and wound complications.Using CPT code alone captures 10% of THA and 4% of TKA patients with procedures for nonroutine primary indications. It is essential to recognize identification of patients simply by CPT code has the potential to inadvertently introduce bias, and surgeons should critically assess methods used to define the study populations.

Background The use of vaginal white blood cell (WBC) counts to predict sexually transmitted infections (STIs) in the emergency department (ED) is incompletely characterized. Objectives Our objective was to assess the relationship between vaginal wet preparation WBC counts and STIs and to determine whether WBC counts of at least 11 WBCs per high-power field (HPF) could be useful for identifying STIs in women in the ED. Methods Female ED patients 18 years or older who were evaluated in a single health system between April 18, 2014, and March 7, 2017, and had a genital wet preparation WBC result were retrospectively examined using univariable and multivariable analysis. Results Vaginal wet preparation WBC counts were examined for 17,180 patient encounters. Vaginal WBC counts of at least 11 WBCs/HPF were associated with increased odds of having gonorrhea, chlamydia, or trichomoniasis. When this threshold was used for the diagnosis of each STI, sensitivity ranged from 48.2% to 53.9%, and specificity ranged from 67.2% to 68.8%. Conclusion Women with STIs are more likely to have higher vaginal WBC counts. However, higher vaginal wet preparation WBC counts in isolation have limited diagnostic utility for gonorrhea, chlamydia, and trichomoniasis. Incorporation of age, urine leukocyte esterase results, and vaginal WBC counts provided a better predictor of an STI than vaginal WBC counts alone.

Introduction Vaginal infections are common in the emergency department (ED) but the frequency of vaginal coinfections identified on wet preparation is unknown. Methods The study examined a data set of 75,000 ED patient encounters between April 18, 2014, and March 7, 2017, who had received testing for gonorrhea, chlamydia, or trichomonas or had received a urinalysis and urine culture during the ED encounter. From this data set we reviewed 16,484 patient encounters where a vaginal wet preparation was performed on women age 18 years and older. Findings from the vaginal wet preparation and ED discharge diagnoses were examined to evaluate the frequency of vaginal coinfections with vulvovaginal candidiasis, trichomoniasis, and bacterial vaginosis. Results Among the women who had wet preparations, 4,124 patient encounters (25.0%) had a diagnosis of bacterial vaginosis, 625 (3.8%) had a diagnosis of vulvovaginal candidiasis, and 1,802 (10.9%) were infected with Trichomonas vaginalis. Twenty encounters (0.1%) had a diagnosis of vulvovaginal candidiasis and trichomoniasis; 150 (0.9%), bacterial vaginosis and trichomoniasis; 136 (0.8%), vulvovaginal candidiasis and bacterial vaginosis; and 10 (0.1%), trichomoniasis, bacterial vaginosis, and vulvovaginal candidiasis. On vaginal wet preparation, the mean white blood cell count was 13.0 per high-power field. Clue cells were found in 6,988 wet preparations (42.4%); 1,065 wet preparations (6.5%) had yeast and 1,377 (8.4%) had T. vaginalis. T. vaginalis was identified in 2.5% (266/10,542) of urinalyses and 8.4% (406/4,821) of nucleic acid amplification tests. Conclusions Vaginal coinfections were uncommon among women receiving a vaginal wet preparation in the emergency department. The most common vaginal coinfection was bacterial vaginosis and trichomonas.

Introduction Sexually transmitted infections are commonly tested for in the emergency department (ED), but diagnostic test results are often unavailable during the clinical encounter. Methods We retrospectively reviewed health records of 3,132 men ≥18 years that had an emergency department visit in northeast Ohio between April 18, 2014 and March 7, 2017. All subjects underwent testing for Neisseria gonorrhoeae and Chlamydia trachomatis. Independent t-tests and chi-square analyses were performed as well as multivariable regression analysis. Results On univariable analysis, men with N gonorrhoeae and/or C trachomatis, compared with uninfected men, were younger (25.9 vs 32.4 years), more likely to be of Black race (91.7% vs 85.6%), less likely to be married (3.7% vs 10.2%), less likely to arrive to the ED by ambulance or police (1.7% vs 4.1%), and more likely to be diagnosed with a urinary tract infection (8.3% vs 3.7%), to be treated for gonorrhea and chlamydia in the ED (84.6% vs 54.9%), and to have higher emergency severity index (ESI) scores (3.8 vs 3.6) (P ≤ .03 for all). On urinalysis, men infected with N gonorrhoeae and/or C trachomatis had significantly more white blood cells (55.1 vs 20.9); more mucus (1.3 vs 1.2); higher leukocyte esterase (1.5 vs .4); fewer squamous epithelial cells (.6 vs 1.4); higher urobilinogen (1.1 vs .8); higher bilirubin (.09 vs .05); and more protein (.4 vs .3) (P ≤ .04). Conclusions Demographic and urinalysis findings can be associated with an increased odds of men being infected with N gonorrhoeae and/or C trachomatis.

The two-detector design of the NOvA neutrino oscillation experiment, in which two functionally identical detectors are exposed to an intense neutrino beam, aids in canceling leading order effects of cross-section uncertainties. However, limited knowledge of neutrino interaction cross sections still gives rise to some of the largest systematic uncertainties in current oscillation measurements. We show contemporary models of neutrino interactions to be discrepant with data from NOvA, consistent with discrepancies seen in other experiments. Adjustments to neutrino interaction models in GENIE that improve agreement with our data are presented. We also describe systematic uncertainties on these models, including uncertainties on multi-nucleon interactions from a newly developed procedure using NOvA near detector data., Comment: Code implementing adjustments to GENIE 2.12.2 described in this paper is available at https://github.com/novaexperiment/NOvARwgt-public

Introduction Prostatitis is one of the most common urologic diseases in ambulatory patients. However, prostatitis data are limited from the emergency department (ED) setting. Methods A data set was examined of patients age 18 years or older who received urinalysis and urine culture or were tested for gonorrhea, chlamydia, or trichomonas in the ED from a health care system in northeast Ohio. Results Of 19,308 ED encounters of male patients, 77 encounters (0.4%) involved the diagnosis of prostatitis. Men with prostatitis were younger (52.4 vs 66.3 years), were less likely to be hospitalized (27.3% vs 43.1%), had shorter clinical encounters (1336.5 vs 3019.3 min), and were less likely to arrive by emergency medical services or police (6.5% vs 45.5%) than men diagnosed with urinary tract infection (UTI) without prostatitis (n = 2527) (P ≤ .007 for all). Of the men with urinalysis, those with prostatitis had less bacteria (0.9+ vs 1.8+), blood (0.9+ vs 1.5+), glucose (4.0% vs 13.0%), leukocyte esterase (0.9+ vs 2.3+), nitrite positive (8.0% vs 21.4%), protein (0.5+ vs 1.2+), squamous epithelial cells (0.6 vs 1.7 per high-power field [HPF]), red blood cells (18.3/HPF vs 29.5/HPF), and white blood cells (31.6/HPF vs 57.6/HPF) than men diagnosed with UTI and no prostatitis (P ≤ .005 for all). Escherichia coli was the most common bacterium growing in the urine (58.8%; n = 10) and the blood (100.0%; n = 2) of men with prostatitis; however 73.0% (n = 17) of urine cultures and 90.9% (n = 22) of blood cultures had no bacterial growth. Of 77 patient encounters with prostatitis, 16 (20.8%) underwent testing for Neisseria gonorrhoeae and Chlamydia trachomatis and 3 (3.9%) for Trichomonas vaginalis. Of those tested, only 1 person was infected, with C trachomatis. Conclusion Prostatitis was uncommonly diagnosed in men undergoing urinalysis and urine culture or testing for sexually transmitted infections in the ED.

There is increasing evidence of a significant correlation between prolonged drug-target residence time and increased drug efficacy. Here, we report a structural rationale for kinetic selectivity between two closely related kinases: focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (PYK2). We found that slowly dissociating FAK inhibitors induce helical structure at the DFG motif of FAK but not PYK2. Binding kinetic data, high-resolution structures and mutagenesis data support the role of hydrophobic interactions of inhibitors with the DFG-helical region, providing a structural rationale for slow dissociation rates from FAK and kinetic selectivity over PYK2. Our experimental data correlate well with computed relative residence times from molecular simulations, supporting a feasible strategy for rationally optimizing ligand residence times. We suggest that the interplay between the protein structural mobility and ligand-induced effects is a key regulator of the kinetic selectivity of inhibitors of FAK versus PYK2.

Kinases represent one of the most intensively pursued groups of targets in modern-day drug discovery. Often it is desirable to achieve selective inhibition of the kinase of interest over the remaining ∼500 kinases in the human kinome. This is especially true when inhibitors are intended to be used to study the biology of the target of interest. We present a pipeline of open-source software that analyzes public domain data to repurpose compounds that have been used in previous kinase inhibitor development projects. We define the dual-specificity tyrosine-regulated kinase 1A (DYRK1A) as the kinase of interest, and by addition of a single methyl group to the chosen starting point we remove glycogen synthase kinase β (GSK3β) and cyclin-dependent kinase (CDK) inhibition. Thus, in an efficient manner we repurpose a GSK3β/CDK chemotype to deliver 8b, a highly selective DYRK1A inhibitor.

We describe SGC-GAK-1 (11), a potent, selective, and cell-active inhibitor of cyclin G-associated kinase (GAK), together with a structurally related negative control SGC-GAK-1N (14). 11 was highly selective in an in vitro kinome-wide screen, but cellular engagement assays defined RIPK2 as a collateral target. We identified 18 as a potent RIPK2 inhibitor lacking GAK activity. Together, this chemical probe set can be used to interrogate GAK cellular biology.

Objectives Far cortical locking (FCL) constructs have been shown to increase axial interfragmentary displacement while limiting shear and have been specifically recommended in the treatment of distal femur fractures. However, there is no available data regarding their mechanical behavior within the range of bridge spans typically used for comminuted distal femur fractures. This biomechanical study of distal femur locked plate fixation assessed 4 methods of diaphyseal fixation for associated axial and shear displacement at bridge spans typically used in clinical practice. Methods Distal femur locking plates were used to bridge simulated fractures in femur surrogates with 4 different methods of diaphyseal fixation (bicortical locking, bicortical nonlocking, near cortical locking, and FCL). Axial and shear displacement were assessed at 5 different bridge spans for each fixation method. Results Diaphyseal fixation type was associated with the amount of shear (P = 0.04), but not the amount of axial displacement (P = 0.39). Specifically, FCL constructs demonstrated greater shear than bicortical locking (median 4.57 vs. 2.94 mm, P = 0.02) and bicortical nonlocking (median 4.57 vs. 3.41 mm, P = 0.02) constructs. Conclusions Unexpectedly, FCL constructs demonstrated greater shear than bicortical locking and nonlocking constructs and similar axial displacement for all fixation methods. Bridge span had a dominant effect on displacement that interacted negatively with more flexible FCL diaphyseal fixation. Potentially interactive construct features are best studied in concert. Given the complexity of these relationships, computational modeling will likely play an integral role in future mechanotransduction research.

Over one hundred bacterial species have been determined to comprise the human microbiota in a healthy individual. Bacteria including Escherichia coli, Bacillus cereus, Clostridioides difficile, and Vibrio parahaemolyticus are found inside of the human body and B. cereus and E. coli are also found on the skin. These bacteria can act as human pathogens upon ingestion of contaminated food or water, if they enter an open wound, or antibiotics, and environment or stress can alter the microbiome. In this study, we present new polymerase chain reaction (PCR) high-resolution melt (HRM) assays to detect and identify the above microorganisms. Amplified DNA from C. difficile, E. coli, B. cereus, and V. parahaemolyticus melted at 80.37 &plusmn, 0.45 &deg, C, 82.15 &plusmn, 0.37 &deg, C, 84.43 &plusmn, 0.50 &deg, C, and 86.74 &plusmn, 0.65 &deg, C, respectively. A triplex PCR assay was developed to simultaneously detect and identify E. coli, B. cereus, and V. parahaemolyticus, and cultured microorganisms were successfully amplified, detected, and identified. The assays demonstrated sensitivity, specificity, reproducibility, and robustness in testing.

Background Spinopelvic pathology has been identified as a potential risk factor for instability after total hip arthroplasty. Spinopelvic radiographic parameters used to diagnose spinopelvic disease may also impact optimal cup placement. The purpose of this study was to assess the arthroplasty surgeon accuracy in making spinopelvic measurements. Methods Five fellowship-trained or in-training arthroplasty surgeons reviewed 44 lateral lumbar radiographs in two sessions. All evaluators were instructed how to perform measurements but had little experience in doing so. Traditional measurements included the pelvic tilt (PT), pelvic incidence (PI), and sacral slope (SS), and novel measurements included the acetabular ante-inclination (AI) and pelvic femoral angle. Surgeon measurements were compared with those made by experienced engineers. intraclass coefficients (ICCs) were calculated and interpreted. Results The interobserver reliability for 4 of 5 of the spinopelvic parameters was good to excellent (ICC >0.75) with the highest reliability seen for PI measurement (ICC = 0.939). Only moderate interobserver reliability was observed for AI measurement (ICC = 0.559). Intraobserver reliability was the highest for the PI and SS, ranging from moderate to good (ICC, 0.718 to 0.896). The lowest intraobserver reliability was seen for the AI (ICC range, 0.026 to 0.545) and pelvic femoral angle (ICC range, 0.035 to 0.828). Surgeon measurements of PT and SS were compared with engineer measurements with extremely poor correlation observed (ICC Conclusion Surgeon intraobserver and interobserver reliability in making novel radiographic measurements was low compared with traditional radiographic measurements. Surgeon reliability in making traditional measurements of the PT and SS was very poor compared with experienced assessors using software-based measurements.

Water networks within kinase inhibitor design and more widely within drug discovery are generally poorly understood. The successful targeting of these networks prospectively has great promise for all facets of inhibitor design, including potency and selectivity on target. Here we describe the design and testing of a targeted library of 4-anilinoquinolines for use as inhibitors of cyclin G associated kinase (GAK). The GAK cellular target engagement assays, ATP binding site modelling and extensive water mapping provide a clear route to access potent inhibitors for GAK and beyond.

The STK32 kinases are a small subfamily of three uncharacterised serine/threonine kinases from the AGC kinase family whose functional role is so far unknown. Here, we analyse the consensus peptide sequence for STK32A phosphorylation, showing that STK32A is directed towards acidic substrate sequences and exhibits dual-specificity for serine/threonine and tyrosine residues. A crystal structure of STK32A reveals an overall structure typical of the AGC protein kinase family but with significant and unique features including an altered binding mode of the hydrophobic motif to the N-terminal lobe of the kinase domain, and a novel alpha-helix in between the turn motif and the hydrophobic motif. The crystal structure combined with phylogenetic analysis reveals the evolutionary conservation of the acidic substrate preference.In vitrobinding assays demonstrated that the STK32 kinases bind significant numbers of clinically used kinase inhibitors.

Background Obesity has previously been demonstrated to be an independent risk factor for increased complications after total hip and knee arthroplasties (THA and TKA). The purpose of this study was to compare the effects of obesity and body mass index (BMI) to determine whether the magnitude of the effect was similar for both procedures. Methods We queried the American College of Surgeons National Surgical Quality Improvement Program database to identify patients who underwent primary THA or TKA between 2010 and 2014. Patients were stratified by procedure and classified as nonobese, obese, or morbidly obese according to BMI. Thirty-day rates of wound complications, deep infection, total complications, and reoperation were compared using univariate and multivariate logistic regression analyses. Results We identified 64,648 patients who underwent THA and 97,137 patients who underwent TKA. Obese THA patients had significantly higher rates of wound complications (1.53% vs 0.96%), deep infection (0.31% vs 0.17%), reoperation rate (2.11% vs 1.02%), and total complications (5.22% vs 4.63%) compared with TKA patients. Morbidly obese patients undergoing THA were also found to have significantly higher rates of wound complications (3.25% vs 1.52%), deep infection (0.84% vs 0.23%), reoperation rate (3.65% vs 1.60%), and total complications (7.36% vs 5.57%). Multivariate regression analysis identified increasingly higher odds of each outcome measure as BMI increased. Conclusion This study demonstrates that the impact of obesity on postoperative complications is more profound for THA than TKA. This emphasizes the importance of considering patient comorbidities in the context of the specific procedure when assessing risks of surgery.

Psilocybe cubensis,or“magic mushroom,” is the most common species of fungus with psychedelic characteristics. Two primer sets were designed to target Psilocybe DNA using web-based software and NBCI gene sequences. DNA was extracted from eighteen samples, including twelve mushroom species, using the Qiagen DNeasy Plant Mini Kit. The DNA was amplified by the polymerase chain reaction (PCR) using the primers and a master mix containing either a SYBR Green I, RadiantTM Green, or LCGreen Plus intercalating dye; amplicon size was determined using agarose gel electrophoresis. The PCR assays were tested for amplifiability, specificity, reproducibility, robustness, sensitivity, and multiplexing with primers that target marijuana. The observed high resolution melt (HRM) temperatures for primer sets 1 and 7 were 78.85 +_ 0.31°C and 73.22 +- 0.61°C, respectively, using SYBR Green I dye and 81.67+- 0.06°C and 76.04 +- 0.11°C, respectively, using RadiantTM Green dye.

Calcium/Calmodulin-dependent Protein Kinase Kinase 2 (CAMKK2) acts as a signaling hub, receiving signals from various regulatory pathways and decoding them via phosphorylation of downstream protein kinases - such as AMPK (AMP-activated protein kinase) and CAMK types I and IV. CAMKK2 relevance is highlighted by its constitutive activity being implicated in several human pathologies. However, at present, there are no selective small-molecule inhibitors available for this protein kinase. Moreover, CAMKK2 and its closest human homolog, CAMKK1, are thought to have overlapping biological roles. Here we present six new co-structures of potent ligands bound to CAMKK2 identified from a library of commercially-available kinase inhibitors. Enzyme assays confirmed that most of these compounds are equipotent inhibitors of both human CAMKKs and isothermal titration calorimetry (ITC) revealed that binding to some of these molecules to CAMKK2 is enthalpy driven. We expect our results to advance current efforts to discover small molecule kinase inhibitors selective to each human CAMKK.

Alterations in the NRF2/KEAP1 pathway result in the constitutive activation of NRF2, leading to the aberrant induction of antioxidant and detoxification enzymes, including NQO1. The NQO1 bioactivatable agent β-lapachone can target cells with high NQO1 expression but relies in the generation of reactive oxygen species (ROS), which are actively scavenged in cells with NRF2/KEAP1 mutations. However, whether NRF2/KEAP1 mutations influence the response to β-lapachone treatment remains unknown. To address this question, we assessed the cytotoxicity of β-lapachone in a panel of NSCLC cell lines bearing either wild-type or mutant KEAP1. We found that, despite overexpression of NQO1, KEAP1 mutant cells were resistant to β-lapachone due to enhanced detoxification of ROS, which prevented DNA damage and cell death. To evaluate whether specific inhibition of the NRF2-regulated antioxidant enzymes could abrogate resistance to β-lapachone, we systematically inhibited the four major antioxidant cellular systems using genetic and/or pharmacologic approaches. We demonstrated that inhibition of the thioredoxin-dependent system or copper-zinc superoxide dismutase (SOD1) could abrogate NRF2-mediated resistance to β-lapachone, while depletion of catalase or glutathione was ineffective. Interestingly, inhibition of SOD1 selectively sensitized KEAP1 mutant cells to β-lapachone exposure. Our results suggest that NRF2/KEAP1 mutational status might serve as a predictive biomarker for response to NQO1-bioactivatable quinones in patients. Further, our results suggest SOD1 inhibition may have potential utility in combination with other ROS inducers in patients with KEAP1/NRF2 mutations., Graphical abstract Image 1, Highlights • Aberrant activation of NRF2 in non-small cell lung cancer promotes resistance to β-lapachone via the antioxidant defense. • Inhibition of the thioredoxin-dependent system and superoxide dismutase 1 increase sensitivity to β-lapachone treatment. • Mutations in the NRF2/KEAP1 pathway might serve as predictive biomarker for response to β-lapachone in patients.

There is increasing evidence of a significant correlation between prolonged drug-target residence time and increased drug efficacy. Here, we report a structural rational for kinetic selectivity between two closely related kinases: Focal Adhesion Kinase (FAK) and Proline-rich Tyrosine Kinase 2 (PYK2). We found that slow off-rate FAK inhibitors induce helical structure at the DFG motif of FAK but not PYK2. Binding kinetic data, high resolution structures and mutagenesis data support the role of hydrophobic interactions of inhibitors with the DFG helical region providing a structural rational for slow dissociation rates from FAK and kinetic selectivity over PYK2. Our experimental data correlated well with computed relative residence times from molecular simulations providing a feasible strategy for rationally optimizing ligand residence times. We suggest that the interplay between the protein structural mobility and ligand-induced effects is a key regulator of the kinetic selectivity of inhibitors of FAK versus PYK2.

[Image: see text] Inhibitors based on a 3-acylaminoindazole scaffold were synthesized to yield potent dual AAK1/BMP2K inhibitors. Optimization furnished a small molecule chemical probe (SGC-AAK1-1, 25) that is potent and selective for AAK1/BMP2K over other NAK family members, demonstrates narrow activity in a kinome-wide screen, and is functionally active in cells. This inhibitor represents one of the best available small molecule tools to study the functions of AAK1 and BMP2K.

The NOvA experiment has made a $4.4\sigma$-significant observation of $\bar\nu_{e}$ appearance in a 2 GeV $\bar\nu_{\mu}$ beam at a distance of 810 km. Using $12.33\times10^{20}$ protons on target delivered to the Fermilab NuMI neutrino beamline, the experiment recorded 27 $\bar\nu_{\mu} \rightarrow \bar\nu_{e}$ candidates with a background of 10.3 and 102 $\bar\nu_{\mu} \rightarrow \bar\nu_{\mu}$ candidates. This new antineutrino data is combined with neutrino data to measure the oscillation parameters $|\Delta m^2_{32}| = 2.48^{+0.11}_{-0.06}\times10^{-3}$ eV$^2/c^4$, $\sin^2 \theta_{23} = 0.56^{+0.04}_{-0.03}$ in the normal neutrino mass hierarchy and upper octant and excludes most values near $\delta_{\rm CP}=\pi/2$ for the inverted mass hierarchy by more than 3$\sigma$. The data favor the normal neutrino mass hierarchy by 1.9$\sigma$ and $\theta_{23}$ values in the upper octant by 1.6$\sigma$., Comment: 8 pages, 3 figures. Supplementary material attached (6 figures). To view attachments, please download and extract the gzipped tar source file listed under "Other formats". Fixed supplementary material to include just the compiled pdf not the Latex Source