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Background Dominant gamma-smooth muscle actin gene ( ACTG2) variants cause clinically diverse forms of visceral myopathy. Many patients undergo intestinal resection or biopsy before identification of their genetic defect. The pathology of ACTG2-variant visceral myopathy has not been evaluated systematically. Methods Glass slides, ultrastructural images, molecular genetic reports, and clinical records from 16 patients with pathogenic (15) or likely pathogenic (1) ACTG2 variants were reviewed and compared with surgical specimens from controls (no evidence of a primary myopathy or pseudo-obstruction due to Hirschsprung disease) and published descriptions. Results The variable clinical manifestations in our cohort matched those in the literature. Only non-specific light and electron microscopic findings observed in non-myopathic controls were encountered in 13 of 16 patients. The remaining 3 patients harbored hyalinized cytoplasmic inclusions in smooth muscle cells and 1 of them had polyglucosan bodies in the muscularis propria. Conclusions Apart from hyalinized inclusions, which were only observed in 3/16 patients, intestinal pathology in the majority of patients with ACTG2 variants is not indicative of an underlying visceral myopathy. Molecular testing should be considered even when no diagnostic intestinal pathology is identified.

Offshore wind energy deployment in the US is expected to increase in the years to come, with proposed wind farm sites located in regions with high-risk for tropical cyclones. Yet, the wind turbine design criteria outlined by the International Electrotechnical Commission for extreme events may not account for the severe wind conditions in tropical cyclones, even the weaker storms that are likely to reach mid-Atlantic wind resource areas. To evaluate if current design standards capture the extreme conditions of these storms, we perform idealized large-eddy simulations of five tropical cyclones (two category-1, two category-2, and one category-3 storms) using the Weather Research and Forecasting model. Wind conditions near the eyewall of category-1, category-2 and category-3 storms can exceed current design standards for offshore wind turbines. Hub-height winds can exceed design criteria for Class I and Class T turbines for 50-year recurrence periods. Moreover, wind speed shear across the turbine rotor layer is larger than assumed in design specifications. Vertical variations in wind direction across the turbine rotor layer are also large for tropical cyclones of all intensity levels, suggesting design standards should include veer, which can amplify loads in wind turbines.

Rain-on-snow (ROS) melt events reduce the amount of water stored in the snowpack while also exacerbating flooding. The hydrologic implications of changing ROS events in a warming climate, however, are still uncertain. This research used a calibrated and validated Soil and Water Assessment Tool (SWAT) hydrologic model, modified with energy budget equations to simulate ROS melt and forced with a climate model ensemble representing moderate greenhouse gas concentrations, to simulate changes to ROS melt in the North American Great Lakes Basin from 1960–2069. The changes to ROS events between the historic period (1960–1999) and mid-century (2040–2069) represent an approximately 30 % reduction in melt in warmer, southern subbasins but less than 5 % reduction in melt in colder, northern subbasins. Additionally, proportionally more rainfall reduces the formation of snowpacks, with area-weighted combined winter and spring rain-to-snow ratios rising from approximately 1.5 historically to 1.9 by the mid-21st century. Areas with historic mean combined winter and spring air temperatures lower than −2 ∘C have ROS regimes that are resilient to mid-21st century warming projections, but ROS occurrence in areas that have mean combined winter and spring temperatures near the freezing point are sensitive to changing air temperatures. Also, relationships between changes in the timing of ROS melt and water yield endure throughout spring but become weak by summer. As the influence of ROS melt events on hydrological systems is being altered in a changing climate, these conclusions are important to inform adaptive management of freshwater ecosystems and human uses in regions of the globe that are sensitive to changes in ROS events.

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Elevated immunity to cancer-expressed antigens can be detected in people with no history of cancer and may contribute to cancer prevention. We have previously reported that MHC-restricted phosphopeptides are cancer-expressed antigens and targets of immune recognition. However, the extent to which this immunity reflects prior or ongoing phosphopeptide exposures was not investigated. In this study, we found that preexisting immune memory to cancer-expressed phosphopeptides was evident in most healthy donors, but the breadth among donors was highly variable. Although three phosphopeptides were recognized by most donors, suggesting exposures to common microbial/infectious agents, most of the 205 tested phosphopeptides were not recognized by peripheral blood mononuclear cells (PBMC) from any donor and the remainder were recognized by only 1 to 3 donors. In longitudinal analyses of 2 donors, effector immune response profiles suggested active reexposures to a subset of phosphopeptides. These findings suggest that the immunogens generating most phosphopeptide-specific immune memory are rare infectious agents or incipient cancer cells with distinct phosphoproteome dysregulations, and that repetitive immunogenic exposures occur in individual donors. Phosphopeptide-specific immunity in PBMCs and tumor-infiltrating lymphocytes from ovarian cancer patients was limited, regardless of whether the phosphopeptide was expressed on the tumor. However, 4 of 10 patients responded to 1 to 2 immunodominant phosphopeptides, and 1 showed an elevated effector response to a tumor-expressed phosphopeptide. As the tumors from these patients displayed many phosphopeptides, these data are consistent with lack of prior exposure or impaired ability to respond to some phosphopeptides and suggest that enhancing phosphopeptide-specific T-cell responses could be a useful approach to improve tumor immunotherapy.

Small-cell lung cancer (SCLC) is an aggressive malignancy in which inhibitors of PARP have modest single-agent activity. We performed a phase I/II trial of combination olaparib tablets and temozolomide (OT) in patients with previously treated SCLC. We established a recommended phase II dose of olaparib 200 mg orally twice daily with temozolomide 75 mg/m2 daily, both on days 1 to 7 of a 21-day cycle, and expanded to a total of 50 patients. The confirmed overall response rate was 41.7% (20/48 evaluable); median progression-free survival was 4.2 months [95% confidence interval (CI), 2.8–5.7]; and median overall survival was 8.5 months (95% CI, 5.1–11.3). Patient-derived xenografts (PDX) from trial patients recapitulated clinical OT responses, enabling a 32-PDX coclinical trial. This revealed a correlation between low basal expression of inflammatory-response genes and cross-resistance to both OT and standard first-line chemotherapy (etoposide/platinum). These results demonstrate a promising new therapeutic strategy in SCLC and uncover a molecular signature of those tumors most likely to respond.Significance:We demonstrate substantial clinical activity of combination olaparib/temozolomide in relapsed SCLC, revealing a promising new therapeutic strategy for this highly recalcitrant malignancy. Through an integrated coclinical trial in PDXs, we then identify a molecular signature predictive of response to OT, and describe the common molecular features of cross-resistant SCLC.See related commentary by Pacheco and Byers, p. 1340.This article is highlighted in the In This Issue feature, p. 1325

Supplementary Tables S3-S8

Table S1. Summary of samples used for this study. Table S2. Antibodies used in the study. Figure S1. Mass spectra of all peptides referenced in figures 2 and 3. Figure S2. ETD Mass spectrum of RPPItQSSL containing an asymmetrically dimethylated Arg residue. Figure S3. ICS flow cytometry plots for HD5. Figure S4. Gating strategy for phenotyping shown in Fig 2E. Figure S5. Healthy donor immunity to O-GlcNAc peptides measured using IFNgamma ELISpot. Figure S6. ICS flow cytometry plots for HD5. Figure S7. Positional analysis of O-GlcNAc peptides.

Clinical trial protocol

Supplementary Figures 1-11

Text for supplement: Supplementary methods, supplementary figure legends

Leukemias are highly immunogenic, but they have a low mutational load, providing few mutated peptide targets. Thus, the identification of alternative neoantigens is a pressing need. Here, we identify 36 MHC class I–associated peptide antigens with O-linked β-N-acetylglucosamine (O-GlcNAc) modifications as candidate neoantigens, using three experimental approaches. Thirteen of these peptides were also detected with disaccharide units on the same residues and two contain either mono- and/or di-methylated arginine residues. A subset were linked with key cancer pathways, and these peptides were shared across all of the leukemia patient samples tested (5/5). Seven of the O-GlcNAc peptides were synthesized and five (71%) were shown to be associated with multifunctional memory T-cell responses in healthy donors. An O-GlcNAc-specific T-cell line specifically killed autologous cells pulsed with the modified peptide, but not the equivalent unmodified peptide. Therefore, these posttranslationally modified neoantigens provide logical targets for cancer immunotherapy. Cancer Immunol Res; 5(5); 376–84. ©2017 AACR.

Supplementary Tables and Figures

Supplementary Tables 4-5

Supplementary Figure S1. AMG 510 demonstrates cooperativity with SHP2 and RTK inhibition

Supplementary Data 3

Supplementary Figure S9. Combined SHP2 and KRASG12C inhibition improves efficacy in vivo.

Supplementary Figure 1

Purpose:KRAS-mutant lung cancers have been recalcitrant to treatments including those targeting the MAPK pathway. Covalent inhibitors of KRAS p.G12C allele allow for direct and specific inhibition of mutant KRAS in cancer cells. However, as for other targeted therapies, the therapeutic potential of these inhibitors can be impaired by intrinsic resistance mechanisms. Therefore, combination strategies are likely needed to improve efficacy.Experimental Design: To identify strategies to maximally leverage direct KRAS inhibition we defined the response of a panel of NSCLC models bearing the KRAS G12C–activating mutation in vitro and in vivo. We used a second-generation KRAS G12C inhibitor, ARS1620 with improved bioavailability over the first generation. We analyzed KRAS downstream effectors signaling to identify mechanisms underlying differential response. To identify candidate combination strategies, we performed a high-throughput drug screening across 112 drugs in combination with ARS1620. We validated the top hits in vitro and in vivo including patient-derived xenograft models.Results:Response to direct KRAS G12C inhibition was heterogeneous across models. Adaptive resistance mechanisms involving reactivation of MAPK pathway and failure to induce PI3K–AKT pathway inactivation were identified as likely resistance events. We identified several model-specific effective combinations as well as a broad-sensitizing effect of PI3K-AKT–mTOR pathway inhibitors. The G12Ci+PI3Ki combination was effective in vitro and in vivo on models resistant to single-agent ARS1620 including patient-derived xenografts models.Conclusions:Our findings suggest that signaling adaptation can in some instances limit the efficacy of ARS1620 but combination with PI3K inhibitors can overcome this resistance.

Purpose:Although KRAS represents the most commonly mutated oncogene, it has long been considered an “undruggable” target. Novel covalent inhibitors selective for the KRASG12C mutation offer the unprecedented opportunity to target KRAS directly. However, prior efforts to target the RAS–MAPK pathway have been hampered by adaptive feedback, which drives pathway reactivation and resistance.Experimental Design:A panel of KRASG12C cell lines were treated with the KRASG12C inhibitors ARS-1620 and AMG 510 to assess effects on signaling and viability. Isoform-specific pulldown of activated GTP-bound RAS was performed to evaluate effects on the activity of specific RAS isoforms over time following treatment. RTK inhibitors, SHP2 inhibitors, and MEK/ERK inhibitors were assessed in combination with KRASG12C inhibitors in vitro and in vivo as potential strategies to overcome resistance and enhance efficacy.Results:We observed rapid adaptive RAS pathway feedback reactivation following KRASG12C inhibition in the majority of KRASG12C models, driven by RTK-mediated activation of wild-type RAS, which cannot be inhibited by G12C-specific inhibitors. Importantly, multiple RTKs can mediate feedback, with no single RTK appearing critical across all KRASG12C models. However, coinhibition of SHP2, which mediates signaling from multiple RTKs to RAS, abrogated feedback reactivation more universally, and combined KRASG12C/SHP2 inhibition drove sustained RAS pathway suppression and improved efficacy in vitro and in vivo.Conclusions:These data identify feedback reactivation of wild-type RAS as a key mechanism of adaptive resistance to KRASG12C inhibitors and highlight the potential importance of vertical inhibition strategies to enhance the clinical efficacy of KRASG12C inhibitors.See related commentary by Yaeger and Solit, p. 1538

Supplementary Figure S7. SHP2 inhibition enhances the efficacy of KRASG12C inhibition.

Supplementary Figure S8. KRASG12C and SHP2 inhibition does not inhibit the growth of signaling of a non-KRASG12C mutant cell line.

Supplementary Figure S3. RTK inhibitors display variable cooperative effects with KRASG12C inhibitors across models.

Supplementary Table 1

Supplementary Figure S4. Combined KRASG12C and RTK inhibition improves efficacy in vivo.

Supplementary Figure S6. SHP2 inhibition enhances suppression of MAPK signaling in the presence of KRASG12C inhibition.

Supplementary Figure S2. KRASG12C inhibitors are subject to adaptive feedback through activation of RTKs.

Supplementary Figure S5. Combined KRASG12C and SHP2 or MAPK inhibition improves in vitro efficacy

Legends to supplementary table, data and figures

We present observations of an extreme radio flare, VT J024345.70-284040.08, hereafter VT J0243, from the nucleus of a galaxy with evidence for historic Seyfert activity at redshift $z=0.074$. Between NRAO VLA Sky Survey observations in 1993 to VLA Sky Survey observations in 2018, VT J0243 rose from a ${\sim}$GHz radio luminosity of $\nu L_\nu \lesssim 10^{38}$ erg s$^{-1}$ to $\nu L_\nu{\sim}10^{40}$ erg s$^{-1}$, and still continues to brighten. The radio spectral energy distribution (SED) evolution is consistent with a nascent jet that has slowed over ${\sim}3000$ days with an average $0.1 < \langle \beta \rangle < 0.6$. The jet is energetic (${\sim}10^{51-52}$ erg), and had a radius ${\sim}0.7$ pc in Dec. 2021. X-ray observations suggest a persistent or evolving corona, possibly associated with an accretion disk, and IR and optical observations constrain any high-energy counterpart to be sub-Eddington. VT J0243 may be an example of a young, off-axis radio jet from a slowly evolving tidal disruption event. Other more mysterious triggers for the accretion enhancement and jet launching are possible. In either case, VT J0243 is a unique example of a nascent jet, highlighting the unknown connection between supermassive black holes, the properties of their accretion flows, and jet launching., Comment: 20 pages, 5 figures, 3 tables. Submitted to ApJ

Multiparent advanced eneration inter-cross (MAGIC) populations improve the precision of quantitative trait loci (QTL) mapping over biparental populations by incorporating increased diversity and opportunities to reduce linkage disequilibrium among variants. Here, we describe the development of a MAGIC B-Line (MBL) population from an inter-cross among 4 diverse founders of grain sorghum [Sorghum bicolor (L.) Moench] across different races (kafir, guinea, durra, and caudatum). These founders were selected based on genetic uniqueness and several distinct qualitative features including panicle architecture, plant color, seed color, endosperm texture, and awns. A whole set of MBL (708 F6) recombinant inbred lines along with their founders were genotyped using Diversity Arrays Technology (DArTseq) and 5,683 single-nucleotide polymorphisms (SNPs) were generated. A genetic linkage map was constructed using a set of polymorphic, quality-filtered markers (2,728 SNPs) for QTL interval-mapping. For population validation, 3 traits (seed color, plant color, and awns) were used for QTL mapping and genome-wide association study (GWAS). QTL mapping and GWAS identified 4 major genomic regions located across 3 chromosomes (Chr1, Chr3, and Chr6) that correspond to known genetic loci for the targeted traits. Founders of this population consist of the fertility maintainer (A/B line) gene pool and derived MBL lines could serve as female/seed parents in the cytoplasmic male sterility breeding system. The MBL population will serve as a unique genetic and genomic resource to better characterize the genetics of complex traits and potentially identify superior alleles for crop improvement efforts to enrich the seed parent gene pool.

Assessing relative performance of different sampling methods used for early detection monitoring (EDM) is a critical step in understanding the likelihood of detecting new non-indigenous species (NIS) in an environment of interest. EDM performance metrics are typically based on the probability of detecting established NIS or rare indigenous species; however, detection probability estimates for these proxies may not accurately reflect survey effectiveness for newly introduced NIS. We used data from three different EDM survey approaches that varied by targeted life-stage (adult–juvenile versus ichthyoplankton), media (physical fish versus environmental DNA), and taxonomic method (morphology-based versus DNA-based taxonomy) to explore relative detection sensitivity for recently introduced white bass (Morone chrysops) and gizzard shad (Dorosoma cepedianum) in the Port of Duluth-Superior, a NIS introduction hot spot within the Laurentian Great Lakes. Detection efficiency, measured by the effort (number of samples) required to achieve 95% probability of detection, differed by EDM approach and species. Also, the relative sensitivity (detection rate) of each survey approach differed by species. For both species, detection in surveys using DNA-based taxonomy was generally as good or better than the adult–juvenile survey using morphology-based taxonomy. While both species appear to have been detected at early stages of invasion, white bass were likely present up to 5 years prior to initial detection, whereas gizzard shad may have been detected in the first year of introduction. We conclude that using complimentary sampling methods can help to balance the strengths and weaknesses of each approach and provide more reliable early detection of new invaders.

Offshore wind is a promising renewable energy source for many coastal populations, predicted to supply as much as 10% of global electricity by 2050. However, the technology has followed a cautious development path, using an onshore-style of turbine and tower, held steadily with an oil-industry type of platform. This approach has avoided major risks but is responsible for high costs, supply chain and infrastructure challenges, and relatively slow innovation due to the difficulties of offshore repair. It may now be appropriate to rethink that approach in search of lower cost and reduced infrastructure requirements. This paper presents ongoing work on a light floating turbine concept that yaws like a ship at anchor, has a shallow draft, widely spaced buoyancy, and high center of gravity (CG), and holds the rotor both upwind and downwind. Rather than seeking to minimize wave disturbances, the design aims to stiffly follow water elevation by using shallow floats designed to prevent heave amplification. Regular and irregular motions of the system are studied for various mass distributions to identify deviations from the intended wave following motion.

Background Ischemic stroke impacts roughly 700,000 patients per year in the United States, yet there are limited options to improve recovery for individuals with a completed stroke and associated neurological deficit. We describe a preliminary clinical trial of a novel non-invasive device that provides peripheral somatosensory stimulation using intermittent jets of air to the affected hand in patients with ischemic stroke. Methods The safety and short-term impact of the use of a non-invasive device providing peripheral somatosensory stimulation were investigated in four patients after mild to moderate stroke. Perfusion MR imaging was performed with the device off and then on in each patient. Serial evaluations of the NIH stroke scale score, Glasgow Outcome Score, and Barthel Index were performed in each case. Results All patients tolerated treatment with the device. No complications related to the use of the device were encountered. A non-blinded neuroradiologist interpreted the MR imaging as demonstrating improved perfusion in three of four cases. All patients demonstrated neurological improvement over the course of treatment. Conclusions It has been suggested that peripheral somatosensory stimulation may improve recovery from stroke. We investigated the use of a novel non-invasive device that delivers intermittent jets of air to the affected forearm and hand. In this small, preliminary series, treatment was well-tolerated in all cases. A larger trial properly empowered to demonstrate clinical benefit may be warranted.