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Resiquimod (R848) is a toll-like receptor 7 and 8 (TLR7/8) agonist with potent antitumor and immunostimulatory activity. However, systemic delivery of R848 is poorly tolerated because of its poor solubility in water and systemic immune activation. In order to address these limitations, we developed an intravenously-injectable formulation with R848 using thermosensitive liposomes (TSLs) as a delivery vehicle. R848 was remotely loaded into TSLs composed of DPPC: DSPC: DSPE-PEG2K (85:10:5, mol%) with 100mM FeSO4 as the trapping agent inside. The final R848 to lipid ratio of the optimized R848-loaded TSLs (R848-TSLs) was 0.09 (w/w), 10-fold higher than the previously-reported values. R848-TSLs released 80% of R848 within 5min at 42°C. These TSLs were then combined with αPD-1, an immune checkpoint inhibitor, and ultrasound-mediated hyperthermia in a neu deletion (NDL) mouse mammary carcinoma model (Her2+, ER/PR negative). Combined with αPD-1, local injection of R848-TSLs showed superior efficacy with complete NDL tumor regression in both treated and abscopal sites achieved in 8 of 11 tumor bearing mice over 100days. Immunohistochemistry confirmed enhanced CD8+ T cell infiltration and accumulation by R848-TSLs. Systemic delivery of R848-TSLs, combined with local hyperthermia and αPD-1, inhibited tumor growth and extended median survival from 28days (non-treatment control) to 94days. Upon re-challenge with reinjection of tumor cells, none of the previously cured mice developed tumors, as compared with 100% of age-matched control mice. The dose of R848 (10μg for intra-tumoral injection or 6mg/kg for intravenous injection delivered up to 4 times) was well-tolerated without weight loss or organ hypertrophy. In summary, we developed R848-TSLs that can be administered locally or systematically, resulting in tumor regression and enhanced survival when combined with αPD-1 in mouse models of breast cancer.

Objective To implement auditory continual feedback into the interface design of a Level 3 automated vehicle and to test whether gaze behavior and reaction times of drivers improved in take-over situations. Background When required to assume manual control in take-over situations, drivers of Level 3 automated vehicles are less likely than conventional drivers to spot potential hazards, and their reaction time is longer. Therefore, it is crucial that the interface of Level 3 automated vehicles will be designed to improve drivers’ performance in take-over situations. Method In two experiments, participants drove a simulated route in a Level 3 automated vehicle for 35 min with one imminent take-over event. Participants’ gaze behavior and performance in an imminent take-over event were monitored under one of three auditory interface designs: (1) Continual feedback. A system that provides verbal driving-related feedback; (2) Persistent feedback. A system that provides verbal driving-related feedback and a persistent beep; and (3) Chatter feedback. A system that provides verbal non-driving-related feedback. Also, there was a control group without feedback. Results Under all three auditory feedback designs, the number of drivers' on-road glances increased compared to no feedback, but none of the designs shortened reaction time to the imminent event. Conclusion Increasing the number of on-road glances during automated driving does not necessarily improve drivers’ attention to the road and their reaction times during take-overs. Application Possible implications for the effectiveness of auditory continual feedback should be considered when designing interfaces for Level 3 automated vehicles.

BACKGROUND Folate is essential for DNA synthesis, DNA repair, cell proliferation, development, and morphogenesis. Folic acid (FA) is a nutritional supplement used to fortify human diets. OBJECTIVES We investigated the effects of dietary FA on early mammary gland (MG) development and hyperplasia. METHODS Study 1: nulliparous female FVB wild-type (WT) mice were fed control (Con; 2 mg FA/kg), deficient (Def; 0 mg FA/kg), excess (Ex; 5 mg FA/kg), or super excess (S-Ex; 20 mg FA/kg) diets for 8 wk before mating to WT or heterozygous FVB/N-Tg[mouse mammary tumor virus long terminal repeat (MMTV)-polyomavirus middle T antigen (PyVT)]634Mul/J (MMTV-PyMT+/-) transgenic males. Dams were fed these diets until they weaned WT or MMTV-PyMT+/- pups, which were fed the dam's diet from postnatal day (PND) 21 to 42. Tissues were collected from female progeny at PNDs 1, 21, and 42. Study 2: Con or Def diets were fed to WT intact females and males from PND 21 to 56, or to ovariectomized females from PND 21 to 77; tissues were collected at PND 56 or 77. Growth of all offspring, development of MGs, MG hyperplasia, supramammary lymph nodes, thymus and spleen, cell proliferation, and expression of MG growth factors were measured. RESULTS Study 1: Ex or S-Ex did not affect postnatal MG development or hyperplasia. The rate of isometric MG growth (PND 1-21) was reduced by 69% in Def female progeny (P

Crucial transitions in cancer-including tumor initiation, local expansion, metastasis, and therapeutic resistance-involve complex interactions between cells within the dynamic tumor ecosystem. Transformative single-cell genomics technologies and spatial multiplex in situ methods now provide an opportunity to interrogate this complexity at unprecedented resolution. The Human Tumor Atlas Network (HTAN), part of the National Cancer Institute (NCI) Cancer Moonshot Initiative, will establish a clinical, experimental, computational, and organizational framework to generate informative and accessible three-dimensional atlases of cancer transitions for a diverse set of tumor types. This effort complements both ongoing efforts to map healthy organs and previous large-scale cancer genomics approaches focused on bulk sequencing at a single point in time. Generating single-cell, multiparametric, longitudinal atlases and integrating them with clinical outcomes should help identify novel predictive biomarkers and features as well as therapeutically relevant cell types, cell states, and cellular interactions across transitions. The resulting tumor atlases should have a profound impact on our understanding of cancer biology and have the potential to improve cancer detection, prevention, and therapeutic discovery for better precision-medicine treatments of cancer patients and those at risk for cancer.

Summary Albino cavefish sleep less than their pigmented siblings. A recent study used CRISPR–Cas9-directed mutation to establish unambiguously that loss of function of a single gene, ocular and cutaneous albinism2, underlies both phenotypes, one morphological, the other behavioral.

SUMMARYPlant roots integrate environmental signals and developmental programs using exquisite spatiotemporal control. This is apparent in the deposition of suberin, an apoplastic diffusion barrier, which regulates the entry and exit of water, solutes and gases, and is environmentally plastic. Suberin is considered a hallmark of endodermal differentiation, but we find that it is absent in the tomato endodermis during normal development. Instead, suberin is present in the exodermis, a cell type that is absent in the model organismArabidopsis thaliana. Here, we uncover genes driving exodermal suberization and describe its effects on drought responses in tomato, unravelling the similarities and differences with the paradigmatic Arabidopsis endodermis. Cellular resolution imaging, gene expression, and mutant analyses reveal loss of this program from the endodermis, and its co-option in the exodermis. Functional genetic analyses of the tomato MYB92 transcription factor and ASFT enzyme demonstrate the importance of exodermal suberin for a plant water-deficit response. Controlling the degree of exodermal suberization could be a new strategy for breeding climate-resilient plants.

Background Overuse injuries of the shoulder and elbow continue to be prevalent in elite baseball pitchers. Pitch velocity has been shown to impact medial elbow torque in adolescent baseball pitchers. However, the determinants of medial elbow torque in professional baseball pitchers are not known. Purpose To determine the influence of pitch type, velocity, and player characteristics on medial elbow torque in professional baseball pitchers. Methods Professional baseball pitchers were recruited for participation. Height, weight, body mass index (BMI), and throwing arm measurements were obtained for all study participants. While wearing a gyroscopic sensor equipped with an accelerometer, participants were instructed to throw a standard, randomized sequence of fastballs, changeups, and curveballs. Elbow torque, arm slot, arm speed, shoulder rotation, and ball velocity were recorded for each pitch. A linear mixed model was used to evaluate the association of pitch type with each pitch parameter, adjusting for pitchers' demographics. Results A total of 12 professional baseball pitchers were included in this study. Among the pitch types, medial elbow torque was significantly higher in fastballs than in curveballs (P = .001). An increased BMI value was independently associated with decreased elbow torque in pitchers (P = .035). Conclusion Fastballs place significantly higher torque on the medial elbow than do curveballs, which is consistent with previous studies done on high school and collegiate populations. Pitchers with a higher BMI experience significantly less torque across the medial elbow.

The serrated neoplasia pathway gives rise to a distinct subgroup of colorectal cancers distinguished by the presence of mutant BRAFV600E and the CpG Island Methylator Phenotype (CIMP). BRAF mutant CRC are commonly associated with microsatellite instability, which have an excellent clinical outcome. However, a proportion of BRAF mutant CRC retain microsatellite stability and have a dismal prognosis. The molecular drivers responsible for the development of this cancer subgroup are unknown. To address this, we established a murine model of BRAFV600E mutant microsatellite stable CRC and comprehensively investigated the exome and transcriptome to identify molecular alterations in signaling pathways that drive malignancy. Exome sequencing of murine serrated lesions (mSL) and carcinomas identified frequent hot spot mutations within the gene encoding β-catenin (Ctnnb1). Immunohistochemical staining of β-catenin indicated that these mutations led to an increase in the presence of aberrant nuclear β-catenin that resulted in gene expression changes in targets of β-catenin transcription. Gene expression profiling identified a significant enrichment for transforming growth factor-β (TGF-β) signaling that was present in mSL and carcinomas. Early activation of TGF-β suggests that this pathway may be an early cue directing mSL to microsatellite stable carcinoma. These findings in the mouse model support the importance of alterations in WNT and TGF-β signaling during the transition of human sessile serrated lesions to malignancy. Keywords: BRAF, Colorectal cancer, WNT, TGF-β, Microsatellite

Reading ability requires the coordination of many cognitive processes to be effective, including spatial attention. Recent functional magnetic resonance imaging (fMRI) evidence from Ekstrand et al. (2019) suggests that lexical reading is more associated with reflexive attentional orienting regions, whereas sublexical reading is more associated with voluntary attentional orienting regions. The current research sought to further examine the neuroanatomical relationship between reading and attention using a novel experimental design in fMRI. Participants performed four hybrid attentional orienting and reading-aloud tasks, where a reflexive or voluntary spatial cue preceded a lexical or sublexical target. Results indicated that lexical reading resulted in greater activation in the right temporoparietal junction, a reflexive orienting region. Sublexical reading resulted in greater activation in the left inferior frontal gyrus, left fusiform and inferior temporal gyrus, and right superior parietal lobule and intraparietal sulcus (voluntary orienting regions). Further, we found an interaction between reading and attention in the middle occipital gyrus. This study provides the most direct evidence to date that lexical and sublexical reading recruit differential attentional orienting regions during single-word reading in skilled readers. Implications for models of reading and attention, as well as for strategic remediation of their dysfunction, are discussed.

Most patients with Cowden syndrome have lesions in the gastrointestinal tract, characterized by multiple polyps of various histologic types in the large bowel, polyps in the upper gastrointestinal tract, and esophageal glycogenic acanthosis. However, pathologists are often unaware of the distinctive polyposis phenotype of Cowden syndrome. In this multicenter study, we report the spectrum of gastrointestinal manifestations in a series of 43 Cowden syndrome patients who had at least one endoscopy. The median age at the first endoscopy was 46 years and 58% were women. In 24 of 29 (83%) tested patients, a pathogenic germline mutation in PTEN was identified. The histology from 199 endoscopy procedures (67 upper gastrointestinal endoscopy and 132 colonoscopies) was reviewed. Hamartomatous polyps of the large bowel were the most common lesions, present in 85% of patients. Hamartomatous polyps showed varied histology, including lymphoid aggregates in 55% of patients, a lipomatous component in 52%, a ganglioneuromatous component in 52%, and a fibrous-rich component in 14%. Polyps with at least two different stromal components were found in 55% of patients. Inflammatory polyps were present in 21% of patients. Conventional adenomas and serrated polyps were identified in 48% and 62% of patients, respectively. In the upper gastrointestinal tract, the most common lesions were esophageal glycogenic acanthosis (37%), gastric hamartomatous polyps (47%), and duodenal hamartomatous polyps (20%). All patients with glycogenic acanthosis who had a colonoscopy had hamartomatous polyps of the large bowel. In five patients, the diagnosis of Cowden syndrome was established after the pathology report raised suspicion for the diagnosis. Pathologists who are aware of the characteristic admixture of lesions in Cowden syndrome can play an essential role in recommending referral to genetic counseling and gene testing. Early diagnosis of Cowden syndrome is important, as these patients and their relatives are at increased risk for developing multiple cancers.

Light rail (LR) is a popular means of public transportation worldwide, in use in more than 380 cities worldwide. LR drivers must have good hazard awareness: the ability to understand the complexity of the traffic environment and anticipate road events. Yet, no study has examined LR drivers' ability to anticipate hazards, and this is the purpose of this study. The experimental group included 28 certified LR drivers from the LR in Jerusalem. The control group included 26 licensed drivers, with no experience in LR driving. Participants observed 18 short video clips of typical LR driving that were filmed from the LR driver's field of view and had to press a response button each time they identified a hazard. Participants' eye movements and button presses were recorded throughout the experiment. In general, LR drivers were better at identifying hazards compared to the control group. Novice LR drivers with less than 1 year of LR driving experience or under training were more likely to respond to hidden hazards and responded much sooner compared to both the experienced LR drivers and Control drivers. The implications are discussed.

Given the preventable morbidity and mortality associated with atrial fibrillation (AF), increased awareness of undiagnosed AF, and advances in mobile electrocardiogram (ECG) technology, there is a critical need to assess the effectiveness of using such technology to routinely screen for AF in clinical practice. VITAL-AF is a pragmatic trial that will test whether screening for AF using a single-lead handheld ECG in individuals 65 years or older during primary care visits will lead to an increased rate of AF detection. The study is a cluster-randomized trial, with 8 primary care practices randomized to AF screening and 8 primary care practices randomized to usual care. We anticipate studying approximately 16,000 patients in each arm. During the 1-year enrollment period, practice medical assistants will screen eligible patients who agree to participate during office visits using a single-lead ECG device. Automated screening results are documented in the electronic health record, and patients can discuss screening results with their provider during the scheduled visit. All single-lead ECGs are overread by a cardiologist. Screen-detected AF is managed at the discretion of the patient’s physician. The primary study end point is incident AF during the screening period. Key secondary outcomes include new oral anticoagulation prescriptions, incident ischemic stroke, and major hemorrhage during a 24-month period following the study start. Outcomes are ascertained based on electronic health record documentation and are manually adjudicated. The results of this pragmatic trial may help identify a model for widespread adoption of AF screening as part of routine clinical practice.

One of the most significant safety concerns regarding Highly-Automated Driving (HAD) is drivers’ ability to regain control of the vehicle safely. Vibro-tactile alerts were already suggested as an effective modality for Take-Over Requests (TOR) in terms of reducing reaction times. However, it is not clear yet whether such alerts should be compatible or incompatible with the location of hazards that might be present when the TOR is initiated. Studies regarding tactile directionality in other domains, and in manual vehicles have found mixed results. It is argued that part of the contradictory evidence may be related to contextual differences between the driving domain and other domains. Thus, this study aimed to test which directional design would be preferable for TORs in time-critical situations. Twenty-seven participants drove a highly-automated vehicle on a highway with two lanes in each travel direction, in a driving simulator. Each participant experienced five TORs in which they were required to take control and divert their vehicle away from an impending hazard that shut down an entire lane and was situated four seconds ahead. The disengagement of the autonomous driver was signaled using a tactile alert. For the first group, the tactile alert was directed towards the hazard (incompatible with the required action), for the second, it was directed away from it (compatible with the required action), and for the control group, the alert was non-directional. Results showed that drivers using the compatible alert reacted faster and more accurately than those using the incompatible alert. Participants using the non-directional alert reacted slower and less accurately than participants in both directional groups. The results contradict previous findings in the manual driving domain, where drivers are faster and more accurate to respond when the alert is compatible with the location of the hazard and not with the direction of the required action. It is argued that these discrepancies stem from the modified HAD driving task demands where drivers are disengaged from the driving task for long periods and are less aware of the driving environment. The implications for the design of autonomous vehicles are discussed.

Recent research has shown a relationship between reading and attention, however the neuroanatomical overlap of these two processes has remained relatively unexplored. Therefore, we sought to investigate the overlapping neural mechanisms of spatial attention and reading using functional magnetic resonance imaging. Participants performed two attentional orienting tasks (reflexive and voluntary), and two overt word-reading tasks (lexical and sublexical). We hypothesized that there would be greater unique activation overlap of reflexive attention with lexical reading, and of voluntary attention with sublexical reading. Results indicated that lexical reading had greater overlapping activation in reflexive orienting areas compared to sublexical reading, suggesting that lexical reading may employ more automatic attentional mechanisms. In contrast, sublexical reading had greater overlapping activation with voluntary attention areas compared to lexical reading, suggesting that phonetic decoding may rely more heavily on voluntary attention. This research broadens our understanding of the neural overlap that underlies the relationship between reading and spatial attention.