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This study examines how Americans’ narratives of pain are connected to social inequalities using more than 1,500 in-depth interviews collected as part of the American Voices Project. Our novel mixed-method approach to these “big (qualitative) data” uses machine learning and qualitative analysis to demonstrate that discussions of physical pain are prevalent across groups, but narrative content varies by demographics including educational background and gender identity. Our findings demonstrate (1) that pain is commonly described as a challenge that shapes everyday life, (2) that pain narratives often invoke morality and distinction, and (3) that narratives diverge around discussions of unequal medical treatment, illness identities, vulnerability and suffering. We discuss the implications of our findings for scholarship on inequality and experience, social scientific methods, and health policy.

Abstract Comprehensive characterization of protein networks in mounted brain tissue represents a major challenge in brain and neurodegenerative disease research. In this study, we develop a simple staining method, called TSWIFT, to iteratively stain pre-mounted formalin fixed, paraffin embedded (FFPE) brain sections, thus enabling high-dimensional sample phenotyping. We show that TSWIFT conserves tissue architecture and allows for relabeling a single mounted FFPE sample more than 10 times, even after prolonged storage at 4 °C. Our results establish TSWIFT as an efficient method to obtain integrated high-dimensional knowledge of cellular proteomes by analyzing mounted FFPE human brain tissue.

ABSTRACT: Introduction: This study evaluates the inter-rater reliability and accuracy of radiographs in assessing healing of osteochondral allografts (OCA) in the knee compared with computed tomography (CT) imaging is currently unknown. Methods: A retrospective review patients who underwent OCA transplantation with postoperative knee radiographs obtained at three and six months, and CT scans obtained at six months was conducted. Three board-eligible orthopaedic surgeons independently graded postoperative radiographs, utilizing the grading scale established by Oladeji et al (2017), and the CT scans utilizing the grading scale established by Gelber et al (2021). Their consensus measurements served as a comparative gold standard. Results: Among 44 prospectively imaged grafts, 13 (29.5%) had radiographs at three months and 28 (63.6%) had radiographs available for evaluation at six months. Assessment of radiographic integration demonstrated low reliability (Intraclass correlation coefficient (ICC): -0.02 to 0.14). On CT scan, this improved to fair reliability (κ = 0.29, P

Summary: In contrast to most hematopoietic lineages, megakaryocytes (MKs) can derive rapidly and directly from hematopoietic stem cells (HSCs). The underlying mechanism is unclear, however. Here, we show that DNA damage induces MK markers in HSCs and that G2 arrest, an integral part of the DNA damage response, suffices for MK priming followed by irreversible MK differentiation in HSCs, but not in progenitors. We also show that replication stress causes DNA damage in HSCs and is at least in part due to uracil misincorporation in vitro and in vivo. Consistent with this notion, thymidine attenuated DNA damage, improved HSC maintenance, and reduced the generation of CD41+ MK-committed HSCs. Replication stress and concomitant MK differentiation is therefore one of the barriers to HSC maintenance. DNA damage-induced MK priming may allow rapid generation of a lineage essential to immediate organismal survival, while also removing damaged cells from the HSC pool.

Imbalances of iron and dopamine metabolism along with mitochondrial dysfunction have been linked to the pathogenesis of Parkinson's disease (PD). We have previously suggested a direct link between iron homeostasis and dopamine metabolism, as dopamine can increase cellular uptake of iron into macrophages thereby promoting oxidative stress responses. In this study, we investigated the interplay between iron, dopamine, and mitochondrial activity in neuroblastoma SH-SY5Y cells and human induced pluripotent stem cell (hiPSC)-derived dopaminergic neurons differentiated from a healthy control and a PD patient with a mutation in the α-synuclein (SNCA) gene. In SH-SY5Y cells, dopamine treatment resulted in increased expression of the transmembrane iron transporters transferrin receptor 1 (TFR1), ferroportin (FPN), and mitoferrin2 (MFRN2) and intracellular iron accumulation, suggesting that dopamine may promote iron uptake. Furthermore, dopamine supplementation led to reduced mitochondrial fitness including decreased mitochondrial respiration, increased cytochrome c control efficiency, reduced mtDNA copy number and citrate synthase activity, increased oxidative stress and impaired aconitase activity. In dopaminergic neurons derived from a healthy control individual, dopamine showed comparable effects as observed in SH-SY5Y cells. The hiPSC-derived PD neurons harboring an endogenous SNCA mutation demonstrated altered mitochondrial iron homeostasis, reduced mitochondrial capacity along with increased oxidative stress and alterations of tricarboxylic acid cycle linked metabolic pathways compared with control neurons. Importantly, dopamine treatment of PD neurons promoted a rescue effect by increasing mitochondrial respiration, activating antioxidant stress response, and normalizing altered metabolite levels linked to mitochondrial function. These observations provide evidence that dopamine affects iron homeostasis, intracellular stress responses and mitochondrial function in healthy cells, while dopamine supplementation can restore the disturbed regulatory network in PD cells.

Background Intravenous albumin has limited indications supported by randomised controlled trials, yet it is often prescribed for indications not supported by evidence.Aim To reduce unnecessary transfusion of albumin.Interventions Under the leadership of a multidisciplinary quality improvement team, evidence-based recommendations were disseminated in tandem with a new electronic order set, an educational strategy, qualitative interviews with prescribers and a return policy change to reduce wastage.Implementation and evaluation Interventions were introduced in a staggered fashion. The primary outcome, appropriate use of albumin, was monitored and quantified using pre-intervention and post-intervention audits. Process measures included statistical process run charts of monthly usage of 5% and 25% albumin and wastage. Data on length of stay (hospital and intensive care), new inpatient starts on kidney replacement and mortality were collected as balancing measures.Results Appropriate albumin usage based on indication increased from 30% to 50% (p

Introduction: Persons experiencing homelessness (PEH) use emergency medical services (EMS) at disproportionately high rates relative to housed individuals due to several factors including disparate access to healthcare. Limited access to care is compounded by higher rates of substance use in PEH. Despite growing attention to the opioid epidemic and housing crisis, differences in EMS naloxone administration by housing status has not been systematically examined. Our objective in this study was to describe EMS administration of naloxone by housing status in the City of Los Angeles. Methods: This was a 12-month retrospective, cross-sectional analysis of electronic patient care reports (ePCRs) for all 9–1–1 EMS incidents attended by the Los Angeles Fire Department (LAFD), the sole EMS agency for the City of Los Angeles during the study period, January-December 2018. During this time, the City had a population of 3,949,776 with an estimated 31,825 (0.8%) PEH. We included in the study individuals to whom LAFD responders had administered naloxone. Housing status is a mandatory field on ePCRs. The primary study outcome was the incidence of EMS naloxone administration by housing status. We used descriptive statistics and logistic regression models to examine patterns by key covariates. Results: There were 345,190 EMS incidents during the study period. Naloxone was administered during 2,428 incidents. Of those incidents 608 (25%) involved PEH, and 1,820 (75%) involved housed individuals. Naloxone administration occurred at a rate of 19 per 1,000 PEH, roughly 44 times the rate of housed individuals. A logistic regression model showed that PEH remained 2.38 times more likely to receive naloxone than their housed counterparts, after adjusting for gender, age, and respiratory depression (odds ratio 2.38, 95% confidence interval 2.15–2.64). The most common impressions recorded by the EMS responders who administered naloxone were the same for both groups: overdose; altered level of consciousness; and cardiac arrest. Persons experiencing homelessness who received naloxone were more likely to be male (82% vs 67%) and younger (41.4 vs 46.2 years) than housed individuals. Conclusion: In the City of Los Angeles, PEH are more likely to receive EMS-administered naloxone than their housed peers even after adjusting for other factors. Future research is needed to understand outcomes and improve care pathways for patients confronting homelessness and opioid use.

Background: Acinar ductal metaplasia (ADM) is among the earliest initiating events in pancreatic ductal adenocarcinoma (PDAC) development.Methods: We developed a novel morphology-based screen using organoids from wildtype and p48Cre/+ (Cre) mice to discover epigenetic modulators that inhibit or reverse pancreatic ADM more effectively than the broad-spectrum HDAC inhibitor trichostatin A (TSA).Results: Of the 144 compounds screened, nine hits and two additional natural product HDAC inhibitors were validated by dose-response analysis. The class I HDAC inhibitors apicidin and FK228, and the histone methyltransferase inhibitor chaetocin demonstrated pronounced ADM inhibition and reversal without inducing significant cytotoxicity at 1 µM. Thioester prodrug class I HDAC inhibitor largazole attenuated ADM while its disulfide homodimer was effective in both ADM inhibition and reversal. Prioritized compounds were validated for ADM reversal in p48Cre/+; LSL-KrasG12D/+ (KC) mouse organoids using both morphological and molecular endpoints. Molecular index analysis of ADM reversal in KC mouse organoids demonstrated improved activity compared to TSA. Improved prodrug stability translated into a stronger phenotypic and molecular response. RNA-sequencing indicated that angiotensinogen was the top inhibited pathway during ADM reversal.Conclusion: Our findings demonstrate a unique epigenetic mechanism and suggest that the phenotypic screen developed here may be applied to discover potential treatments for PDAC.

In a previous research effort by this group, pseudo engine dynamometer data in multi-dimensional arrays were combined with dynamic equations to form a crank angle resolved engine model compatible with a real-time simulator. The combination of the real-time simulator and external targets enabled the development of a software-in-the-loop (SIL) environment that enabled near-real-time development of AI/ML and real-time deployment of the resulting AI/ML. Military applications, in particular, are unlikely to possess large quantities of non-sparse operational data that span the full operational range of the system, stove-piping the ability to develop and deploy AI/ML which is sufficient for near-real-time or real-time control. AI/ML has been shown to be well suited for predicting highly non-linear mathematical phenomena and thus military systems could potentially benefit from the development and deployment of AI/ML acting as soft sensors. Given the non-sparse nature of the data, it becomes exceedingly important that AI/ML be developed and deployed in near-real-time or real-time in parallel to a real-time system to overcome the inadequacy of applicable data. This research effort used parallel processing to reduce the training duration of the shallow artificial neural networks (SANN) and forest algorithms forming ensemble models. This research is novel in that the SIL environment enables pre-developed AI/ML to be adapted in near-real-time or develop AI/ML in response to changes within the operation of the applied system, different load torques, engine speeds, and atmospheric conditions to name a few. Over time it is expected that the continued adaptation of the algorithms will lead to the development of AI/ML that is suitable for real-time control and energy management.

Background:. We aimed to identify how Integra bilayer wound matrix has expanded facial reconstruction options after Mohs surgery due to its reliability in both single- and dual-stage reconstruction. Methods:. A retrospective review of patients undergoing Mohs surgery and alloplastic facial reconstruction with Integra between 2012 and 2022 was performed. Patients who underwent single-stage reconstruction and dual-stage reconstruction with skin graft with at least 90 days of follow-up were included. Results:. One hundred thirty patients with a median age of 76 years were included. Basal cell carcinoma was the most common malignancy (39%). One hundred forty-two lesions were treated and reconstructed same-day with Integra. Lesions most commonly involved the nose (34%) and forehead (22%). The mean postoperative defect size was 26.9 cm2. An estimated 45.5% (n = 60) of defect sites underwent single-stage reconstruction with healing by secondary intention, whereas 54.5% (n = 72) underwent dual-stage reconstruction with skin graft. Integra success rate was 90.2%. Average time to re-epithelialization was 32.2 + 7.3 days. Average time to repigmentation was 169.5 + 14.6 days. The complication rate was 12.8% (n = 17), with 12 undergoing debridement, three needing new Integra graft, and seven needing new skin grafts. Average size for successful healing without complication was 26.6 cm2. Nineteen sites (13.2%) underwent aesthetic improvement procedures, with the majority occurring after dual-stage reconstruction (n = 13). Conclusions:. Integra is a reliable outpatient reconstructive option for facial Mohs defects that can increase the threshold for autologous tissue harvesting and successfully reconstruct large defects of 26.6 cm2 on average with low complication and reoperation rates.

Summary: Background: While vaccines have established utility against COVID-19, phase 3 efficacy studies have generally not comprehensively evaluated protection provided by previous infection or hybrid immunity (previous infection plus vaccination). Individual patient data from US government-supported harmonized vaccine trials provide an unprecedented sample population to address this issue. We characterized the protective efficacy of previous SARS-CoV-2 infection and hybrid immunity against COVID-19 early in the pandemic over three-to six-month follow-up and compared with vaccine-associated protection. Methods: In this post-hoc cross-protocol analysis of the Moderna, AstraZeneca, Janssen, and Novavax COVID-19 vaccine clinical trials, we allocated participants into four groups based on previous-infection status at enrolment and treatment: no previous infection/placebo; previous infection/placebo; no previous infection/vaccine; and previous infection/vaccine. The main outcome was RT-PCR-confirmed COVID-19 >7–15 days (per original protocols) after final study injection. We calculated crude and adjusted efficacy measures. Findings: Previous infection/placebo participants had a 92% decreased risk of future COVID-19 compared to no previous infection/placebo participants (overall hazard ratio [HR] ratio: 0.08; 95% CI: 0.05–0.13). Among single-dose Janssen participants, hybrid immunity conferred greater protection than vaccine alone (HR: 0.03; 95% CI: 0.01–0.10). Too few infections were observed to draw statistical inferences comparing hybrid immunity to vaccine alone for other trials. Vaccination, previous infection, and hybrid immunity all provided near-complete protection against severe disease. Interpretation: Previous infection, any hybrid immunity, and two-dose vaccination all provided substantial protection against symptomatic and severe COVID-19 through the early Delta period. Thus, as a surrogate for natural infection, vaccination remains the safest approach to protection. Funding: National Institutes of Health.

Viral replication places oncolytic viruses (OVs) in a unique niche in the field of drug pharmacokinetics (PK) as their self-amplification obscures exposure-response relationships. Moreover, standard bioanalytical techniques are unable to distinguish the input from replicated drug products. Here, we combine two novel approaches to characterize PK and biodistribution (BD) after systemic administration of vesicular stomatitis virus pseudotyped with lymphocytic choriomeningitis virus glycoprotein (VSV-GP) in healthy mice. First: to decouple input drug PK/BD versus replication PK/BD, we developed and fully characterized a replication-incompetent tool virus that retained all other critical attributes of the drug. We used this approach to quantify replication in blood and tissues and to determine its impact on PK and BD. Second: to discriminate the genomic and antigenomic viral RNA strands contributing to replication dynamics in tissues, we developed an in situ hybridization method using strand-specific probes and assessed their spatiotemporal distribution in tissues. This latter approach demonstrated that distribution, transcription, and replication localized to tissue-resident macrophages, indicating their role in PK and BD. Ultimately, our study results in a refined PK/BD profile for a replicating OV, new proposed PK parameters, and deeper understanding of OV PK/BD using unique approaches that could be applied to other replicating vectors.

Background:. Breast implant illness (BII) is a poorly understood heterogeneous disorder treated with implant removal; however, patient-reported symptoms and outcomes after treatment remain unclear. Methods:. A retrospective review of patients undergoing bilateral breast implant removal related to BII by two surgeons at an academic medical center between 2018 and 2022 was conducted. Patients were surveyed using the BREAST-Q Reconstruction model with the American Society for Aesthetic Plastic Surgery BII survey extension. Outcomes were analyzed using multivariable logistic regression, adjusted for patient-associated factors. Results:. Forty-seven patients were surveyed with a response rate of 51% (n = 24). Of the 20 patients who completed the survey, the majority were White (85%), with 45% (n = 9) having a documented history of psychiatric illness. Six (30%) patients had capsular contracture and four (20%) had documented implant rupture. Most implant removal procedures (n = 12, 60%) were not covered by insurance. Fourteen (70%) patients reported a net improvement in their symptoms after implant removal, most commonly chest discomfort, muscle pain, fever, and headaches. Capsular contracture was predictive of reduced psychosocial, sexual, and breast satisfaction scores (P = 0.015). Self-pay was predictive of increased breast satisfaction scores (P = 0.009), but had no impact on symptomatic improvement. A reduced time to implant removal was predictive of fewer residual symptoms (P = 0.032). Psychiatric illness had no significant impact on the outcomes. Conclusions:. In the setting of suspected or diagnosed BII, a reduced time to implant removal may decrease the risk of residual symptoms and improve overall patient satisfaction. In patients with capsular contracture, preoperative counseling should emphasize that implant removal may only improve physical symptoms.

Background: As artificial intelligence (AI) continues to advance with breakthroughs in natural language processing (NLP) and machine learning (ML), such as the development of models like OpenAI’s ChatGPT, new opportunities are emerging for efficient curation of electronic health records (EHR) into real-world data (RWD) for evidence generation in oncology. Our objective is to describe the research and development of industry methods to promote transparency and explainability.Methods: We applied NLP with ML techniques to train, validate, and test the extraction of information from unstructured documents (e.g., clinician notes, radiology reports, lab reports, etc.) to output a set of structured variables required for RWD analysis. This research used a nationwide electronic health record (EHR)-derived database. Models were selected based on performance. Variables curated with an approach using ML extraction are those where the value is determined solely based on an ML model (i.e. not confirmed by abstraction), which identifies key information from visit notes and documents. These models do not predict future events or infer missing information.Results: We developed an approach using NLP and ML for extraction of clinically meaningful information from unstructured EHR documents and found high performance of output variables compared with variables curated by manually abstracted data. These extraction methods resulted in research-ready variables including initial cancer diagnosis with date, advanced/metastatic diagnosis with date, disease stage, histology, smoking status, surgery status with date, biomarker test results with dates, and oral treatments with dates.Conclusion: NLP and ML enable the extraction of retrospective clinical data in EHR with speed and scalability to help researchers learn from the experience of every person with cancer.

A no-till experiment was conducted in Auburn, AL U.S.A. to evaluate the effectiveness of an experimental two-stage roller/crimper in reoccurring rolling over the same area planted with a cereal rye/crimson clover cover crop mix and its influence on bush bean yield. Cover crop termination was much greater with rolling/crimping when compared to the non-rolled (untreated) control. During the three growing seasons, rolling three times had significantly higher termination rates compared to all other treatments, exceeding 90% in 2020. These results suggest that there may be an advantage to rolling/crimping three times so that planting of the cash crop could potentially be performed one week earlier, under favorable soil moisture conditions. However, for growing seasons 2018 and 2020 at three weeks after rolling, there were no differences between rolling treatments. In 2019, rolling three times over the same cover crop area was the only treatment that achieved above 90% termination rate indicating a clear advantage of recurring rolling/crimping in 2019. Rolling/crimping proved to be effective as yield was significantly higher compared to not rolled when averaged over all three growing seasons. This is possible due to the difficulty in planting into a standing cover crop which could have negative effects on seed to soil contact, but more importantly explained with the slight soil moisture advantage given to the rolled plots over the standing cover crop plots. Thus, optimum soil moisture when planting beans is key for successful germination and good main crop stand.

ABSTRACT In response to the host environment, the human pathogen Cryptococcus neoformans must rapidly reprogram its translatome from one which promotes growth to one which is responsive to host stress. In this study, we investigate the two events which comprise translatome reprogramming: the removal of abundant, pro-growth mRNAs from the translating pool, and the regulated entry of stress-responsive mRNAs into the translating pool. Removal of pro-growth mRNAs from the translating pool is controlled primarily by two regulatory mechanisms, repression of translation initiation via Gcn2, and decay mediated by Ccr4. We determined that translatome reprogramming in response to oxidative stress requires both Gcn2 and Ccr4, whereas the response to temperature requires only Ccr4. Additionally, we assessed ribosome collision in response to host-relevant stress and found that collided ribosomes accumulated during temperature stress but not during oxidative stress. The phosphorylation of eIF2α that occurred as a result of translational stress led us to investigate the induction of the integrated stress response (ISR). We found that eIF2α phosphorylation varied in response to the type and magnitude of stress, yet all tested conditions induced translation of the ISR transcription factor Gcn4. However, Gcn4 translation did not necessarily result in canonical Gcn4-dependent transcription. Finally, we define the ISR regulon in response to oxidative stress. In conclusion, this study begins to reveal the translational regulation in response to host-relevant stressors in an environmental fungus which is capable of adapting to the environment inside the human host. IMPORTANCE Cryptococcus neoformans is a human pathogen capable of causing devastating infections. It must rapidly adapt to changing environments as it leaves its niche in the soil and enters the human lung. Previous work has demonstrated a need to reprogram gene expression at the level of translation to promote stress adaptation. In this work, we investigate the contributions and interplay of the major mechanisms that regulate entry of new mRNAs into the pool (translation initiation) and the clearance of unneeded mRNAs from the pool (mRNA decay). One result of this reprogramming is the induction of the integrated stress response (ISR) regulon. Surprisingly, all stresses tested led to the production of the ISR transcription factor Gcn4, but not necessarily to transcription of ISR target genes. Furthermore, stresses result in differential levels of ribosome collisions, but these are not necessarily predictive of initiation repression as has been suggested in the model yeast.

Summer cover crop utilization by no-till vegetable farms is essential for continuous soil protection, especially in the southern United States where intense storms are likely to occur in hot and humid summer months. A field experiment was conducted at the National Soil Dynamics Laboratory in Auburn, AL, USA, between the summers of 2015 and 2017 to determine the effectiveness of an experimental roller/crimper in mechanically terminating summer cover crops. Iron clay peas (Vigna unguiculata, L.) planted on a sandy loam and pearl millet (Penninsetum glaucum, L.) planted on clay soil were selected to determine termination rate effectiveness in single, double, and triple rolling/crimping over the same area. Overall, termination rates for both cover crops were higher for rolling three times (71%) compared to rolling once (55%) or twice (63%). However, cover crop termination was inhibited due to rainfalls on the experimental area during the three-week evaluation period. In 2016, drought conditions and high temperatures (32.6 °C) caused biomass reduction, especially for pearl millet, of over 31% to 39% compared to 2017 and 2015. Rolling provided higher soil-water conservation compared with the non-rolled control due the cover crop mulch layer blocking sunlight, keeping the soil surface cooler and preventing water evaporation. Recurrent rolling did not cause soil compaction above the 2.0 MPa level that inhibits root growth, but changes in soil strength were dependent on the soil moisture content.

Producers are looking for more sustainable methods of producing fresh vegetables. No-till systems that use cover crops to keep the soil surface covered can increase sustainability by reducing soil erosion, reducing weed pressure, and retaining soil moisture compared to bare soil cropping systems. Cover crop termination methods along with planting equipment modifications required to work in high residue planting systems are often areas of interest to farmers prior to adopting no-till systems. The objectives of this experiment are to compare the effects of two different cover crops, iron clay pea (Vigna unguiculata L.), and pearl millet (Pennisetum glaucum L.), using three different termination methods (rolled/crimped, mowed, and mowed + incorporated via tillage) and two no-till transplanter configurations (with and without residue manager) on soil properties and collard (Brassica oleracea L. var. viridis) yield. Results show that biomass production for pearl millet was greater, with a three-season average of 8461 kg ha−1 compared to 6465 kg ha−1 for iron clay pea. Rolled cover crops under drier weather conditions retained more volumetric soil moisture compared to other methods for both cover crops. Increases in total soil carbon and nitrogen concentration in the top 15 cm ranged from 8 to 11% and 23 to 35%, respectively. Collards grown in an iron clay pea cover crop produced greater weight and height per plant for all growing seasons compared to those grown in pearl millet. Collard yield was affected by cover crop type as reported with iron clay peas ranging from 2881–13,168 kg ha−1 compared to lower yield for the pearl millet cover crop with a range of 1354–8943 kg ha−1. Overall, both cover crops showed the ability to improve soil health by increasing total soil carbon, but collard production was better in the iron clay pea cover crop.

The objective of this research was to explore biocide occurrence in drinking water resources of Honduras. We collected 46 samples from seven drinking water treatment plants (DWTPs) in Honduras during eight sampling events between October 2018 and August 2019. We used high-resolution mass spectrometry to quantify the concentrations of 55 target biocides and estimate the abundance of four priority suspect biocides and five additional high-abundance biocides. We measured 30 of the target biocides, one of the priority suspect biocides, and all five of the high-abundance biocides in at least one of the samples. No correlation was observed between the overall extent of agriculture and biocide occurrence. However, bean production was strongly and significantly correlated with the biocide mixture complexity, as well as with concentrations of aminobenzimidazole and abscisic acid. Biocide mixture complexity was higher during the rainy season than during the dry season, but biocide concentrations were lower during the rainy season. Finally, we found that existing DWTPs are not consistently effective at removing the target biocides. These data represent the first known broad survey of bioicides in drinking water resources of Honduras and demonstrate the need for further study to better understand and manage biocide occurrence. HIGHLIGHTS Agricultural biocides are measured in water samples from Honduras.; Thirty different target biocides are measured in at least one sample.; The extent of biocide occurrence is not associated with the extent of land used for agriculture.; More biocides measured in water samples from the rainy season but at lower concentrations.; Existing drinking water treatment practices have little effect on biocide concentration.;

Abstract Novel technologies, such as camera traps, have expanded the opportunities for species detection, especially for rare species. Corresponding changes in data processing must occur to handle the large volume of data gathered from technology like camera traps. Automated image data processing, usually by running images through different types of computer algorithms, is an overarching goal to reduce the number of images that researchers must manually review. However, differences in camera trap setups and species characteristics can make automatic processing a challenge. Here, we evaluated the detection accuracy and efficiency of a time‐lapse triggered camera trapping technique combined with a pixel change detection algorithm as part of a monitoring program for a translocated population of the rare and federally threatened Louisiana Pinesnake (Pituophis ruthveni). We paired 5 cameras with automated pit tag readers to collect observations of P. ruthveni. We evaluated an image dataset of 1,500,187 images, collected over 7 months, both manually (i.e., researchers looking at each individual picture to determine snake presence) and automatically using a change detection algorithm. There were 18 P. ruthveni observations recorded by the tag readers, 7 of which occurred while a paired camera was not operational. Ten of the tag reader P. ruthveni observations were captured by the paired camera trap, with an additional P. ruthveni observation from a paired camera trap not recorded by the tag reader. There were 132 snake observations of 13 additional species and 18 observations of unknown snakes from the camera traps. The algorithm reduced the number of images reviewers evaluated by an average of 78.5% per camera (range = 37.3–98.7%) but had a 54.5% success rate at detecting observations of P. ruthveni (47.1% for individual images), and a slightly lower 48.9% success rate detecting other large snakes. Large snakes were 4 times more likely to be flagged by the algorithm than small snakes. Our time‐lapse triggered camera trapping technique performed well with respect to P. ruthveni detection accuracy, compared to the tag readers. However, further research is needed to improve quality assurances of camera trap image filtering and object recognition algorithms across different sites or environments.

Background Recurrent laryngeal nerve (RLN) injury carries significant morbidity. Microsurgical repair of the RLN has proven promising for enhancing patient recovery of vocal function; however, data remains limited. Methods This retrospective cohort study included patients who underwent RLN repair from 2007 to 2022. Demographics and medical history were collected. The location and etiology of RLN injury, as well as the repair technique, were collected. Follow-up data were collected at the initial postoperative visit, at 6 months and at 1 year. Hoarseness was classified as mild, moderate, or severe. Of patients who underwent nasopharyngolaryngoscopy (NPL) following repair, the glottic gap was measured. Vocal interventions performed were also recorded. This study utilized descriptive statistical methods. Results Eleven patients underwent RLN repair. All patients underwent immediate repair. Fifty-four percent (n = 6) of RLN injuries resulted from tumor inflammation or nerve encasement. Eighty-two percent (n = 9) underwent direct RLN coaptation, 9% (n = 1) underwent vagus-RLN anastomosis, and 9% (n = 1) underwent an interposition nerve graft. Technical success was 100%. Seventy-three percent (n = 8) required otolaryngology referral, and of those, 50% (n = 4) required intervention. At initial evaluation, 91% (n = 10) suffered from mild to severe hoarseness, and of patients who underwent NPL, all had a glottic gap. At 1 year, 82% of patients (n = 9) improved to having mild to no appreciable hoarseness. Of the patients who underwent NPL, 62% (n = 5) had closure of the glottic gap. Conclusion Patients undergoing repair of the RLN following injury showed excellent recovery of vocal function and resolution of glottic gap at 1 year.

Changes in cell state underlie the difference between health and disease. Here, the authors propose a computational framework for the integration of gene expression and chromatin-accessibility data from single cells to identify differences in gene regulation in cell types across two conditions.

Visual processing necessitates both extracting and discarding information. Here, the authors use a specialized set of stimuli and two complementary discrimination tasks to demonstrate the opposing perceptual implications of these two aspects of information processing.

Objective: To quantify the occurrence rate of abnormal electrocardiographic (ECG) findings and symptoms following coronavirus disease 2019 (COVID-19) infection. Patients and Methods: In this retrospective analysis, we studied adult patients (>18 years old) who were participating in collegiate athletics and previously tested positive for COVID-19 between August 1, 2020, and December 30, 2020. The athletes underwent general examinations and ECG screening prior to being medically cleared for a return to sports following their COVID-19 diagnosis. Predetermined predictors were grouped into categorical variables including (1) sex, (2) symptom severity, and (3) body mass index (normal vs overweight [≥24 kg/m2]). These variables were used to examine differences of abnormal rates that occurred between different predictor categories. Results: Of the 170 athletes screened, 6 (3.5%) presented with abnormal ECG findings and were referred to cardiologists. We found no evidence that sex, symptom severity, and body mass index category were associated with a higher rate of abnormal ECG findings (all P>.05). Greater severity of COVID-19 symptoms was associated with a higher percentage of ST depression, T-wave inversion, ST-T changes, and the presence of fragmented QRS complex. Loss of smell, loss of taste, headache, and fatigue were the most prevalent symptoms, with 38.8% (66), 36.5% (62), 32.9% (56), and 25.3% (43), respectively, of the 170 athletes reporting each symptom. Conclusion: Preliminary findings indicate a low risk of myocardial injury secondary to COVID-19 infection, with less than 4% of the 170 patients in our study presenting with abnormal ECG findings and a total of 16 patients (9.4%) requiring referral to a cardiologist. Although viral myocarditis was not detected in any athlete referred for cardiological assessment, 2 patients experienced effusive viral pericarditis.

OBJECTIVES/GOALS: People experiencing homelessness vs. housed peers have higher rates of substance use disorders as well as increased emergency medical services (EMS) use. However, it is unknown if EMS administers naloxone at different rates to people experiencing homelessness. We address if patient housing status impacts naloxone administration by EMS providers. METHODS/STUDY POPULATION: We conducted a retrospective, cross-sectional analysis of electronic patient care reports (ePCRs) for all 911-incidents attended by the Los Angeles Fire Department (LAFD) during the study period, January to December 2018. Individuals who were medically assessed by the LAFD and who were administered naloxone by LAFD EMS were included. Exclusion criteria was incomplete ePCRs. The primary outcome was prevalence of EMS naloxone administration by housing status in the city of Los Angeles. We used descriptive statistics and a logistic regression model to examine differences in care. RESULTS/ANTICIPATED RESULTS: Naloxone was administered in 2,438 of the 345,190 incidents that occurred during the study period. 608 (25%) incidents involved people experiencing homelessness. Top indications for naloxone administration were similar in both groups: overdose, altered consciousness and cardiac arrest. Of those who received naloxone, people experiencing homelessness were more likely to be male (82% v 67%) and younger (41 v 46 years). People experiencing homelessness were more likely to receive naloxone (OR 2.6, 95% CI 2.4-2.9). People experiencing homelessness received naloxone at a rate of 44 times that of housed peers. A logistic regression model adjusting for gender, age, respiratory depression and transport status showed people experiencing homelessness remained more likely to receive naloxone (OR 2.3, 95% CI 2.0-2.5). DISCUSSION/SIGNIFICANCE: Emergency medical services are more likely to administer naloxone to people experiencing homelessness than housed peers. There is a need to identify bias and factors that impact prehospital care and patient outcomes of people experiencing homelessness. New care pathways for people confronting homelessness and opioid use disorders are needed.

The last major review of progress toward a chemical retinal prosthesis was a decade ago. Many important advancements have been made since then with the aim of producing an implantable device for animal testing. We review that work here discussing the potential advantages a chemical retinal prosthesis may possess, the spatial and temporal resolutions it might provide, the materials from which an implant might be constructed and its likely effectiveness in stimulating the retina in a natural fashion. Consideration is also given to implant biocompatibility, excitotoxicity of dispensed glutamate and known changes to photoreceptor degenerate retinas.

Background:. Drains are used in plastic surgery to remove excess fluid while ameliorating complications. However, there is a paucity of evidence supporting guiding parameters on when to discontinue a drain. The aim of our study was to determine whether two of the most common parameters, drain volume 24 hours before removal or postoperative day, are valid indicators for drain removal. Methods:. A retrospective chart review was conducted for surgical operations performed by our division between July 2014 and May 2019. Of the 1308 patients, 616 had a drain and a complete record. Demographics, medical history, operative time, antibiotic use, anatomic site, donor/recipient, and complication type were recorded. Complications were defined as events that deviated from expected postoperative course or required pharmacological/procedural intervention. T-test and Chi square were used to analyze data. Results:. In total, 544 patients were in the no complication group, and 72 were in the complication group. The complication group patients had drains removed later than patients in the no complication group (15.7 days versus 12.5 days, P = 0.0003) and had similar final 24-hour drain volumes versus patients in the no complication group (16.7 mL versus 18.8 mL, P = 0.2548). The complication group had more operations on the pelvis (11% versus 2.1%; P = 0.000017) or thigh (8.5% versus 3.4%; P = 0.029). Conclusions:. Our data suggest neither postoperative day nor 24-hour volume before drain removal are valid indicators for removal. Late removal correlates with more complications; however, persisting output leading to later removal may be predictive of an impending complication rather than delays in drain removal causing the complication.

Abstract Background One of the tasks in the iDASH Secure Genome Analysis Competition in 2018 was to develop blockchain-based immutable logging and querying for a cross-site genomic dataset access audit trail. The specific challenge was to design a time/space efficient structure and mechanism of storing/retrieving genomic data access logs, based on MultiChain version 1.0.4 ( https://www.multichain.com/ ). Methods Our technique uses the MultiChain stream application programming interface (which affords treating MultiChain as a key value store) and employs a two-level index, which naturally supports efficient queries of the data for single clause constraints. The scheme also supports heuristic and binary search techniques for queries containing conjunctions of clause constraints, and timestamp range queries. Of note, all of our techniques have complexity independent of inserted data set size, other than the timestamp ranges, which logarithmically scale with input size. Results We implemented our insertion and querying techniques in Python, using the MultiChain library Savoir ( https://github.com/dxmarkets/savoir ), and comprehensively tested our implementation across a benchmark of datasets of varying sizes. We also tested a port of our challenge submission to a newer version of MultiChain (2.0 beta), which natively supports multiple indices. Conclusions We presented creative and efficient techniques for storing and querying log file data in MultiChain 1.0.4 and 2.0 beta. We demonstrated that it is feasible to use a permissioned blockchain ledger for genomic query log data when data volume is on the order of hundreds of megabytes and query times of dozens of minutes is acceptable. We demonstrated that evolution in the ledger platform (MultiChain 1 to 2) yielded a 30%-40% increase in insertion efficiency. All source code for this challenge has been made available under a BSD-3 license from https://github.com/sandialabs/idash2018task1/ .

Abstract Measurement of intracranial pressure (ICP) is crucial in the management of many neurological conditions. However, due to the invasiveness, high cost, and required expertise of available ICP monitoring techniques, many patients who could benefit from ICP monitoring do not receive it. As a result, there has been a substantial effort to explore and develop novel noninvasive ICP monitoring techniques to improve the overall clinical care of patients who may be suffering from ICP disorders. This review attempts to summarize the general pathophysiology of ICP, discuss the importance and current state of ICP monitoring, and describe the many methods that have been proposed for noninvasive ICP monitoring. These noninvasive methods can be broken down into four major categories: fluid dynamic, otic, ophthalmic, and electrophysiologic. Each category is discussed in detail along with its associated techniques and their advantages, disadvantages, and reported accuracy. A particular emphasis in this review will be dedicated to methods based on the use of transcranial Doppler ultrasound. At present, it appears that the available noninvasive methods are either not sufficiently accurate, reliable, or robust enough for widespread clinical adoption or require additional independent validation. However, several methods appear promising and through additional study and clinical validation, could eventually make their way into clinical practice.

How does the brain represent sensory uncertainty? The authors find that neural gain variability tracks stimulus uncertainty across the visual hierarchy and explain their findings with a simple generalization of canonical models of neural computation.

Abstract Background Asthma is a heterogenous disease that can be classified into eosinophilic (type 2‐high) and noneosinophilic (type 2‐low) endotypes. The type 2‐low endotype of asthma can be characterized by the presence of neutrophilic airway inflammation that is poorly responsive to corticosteroids. Dysregulated innate immune responses to microbial products including Toll‐like receptor (TLR) ligands have been associated with the pathogenesis of neutrophilic asthma. The key molecules that regulate inflammatory responses in individuals with neutrophilic asthma remain unclear. We previously reported that the immunoregulatory receptor neuropilin‐2 (NRP2) is expressed by murine and human alveolar macrophage (AM) and suppresses lipopolysaccharide (LPS)‐induced neutrophilic airway inflammation. Methods Here, we investigated the immunoregulatory role of NRP2 in a mouse model of neutrophilic asthma. Results We found that TLR ligands, but not T helper 2 (Th2)‐promoting adjuvants, induced NRP2 expression by AM. Using an LPS‐mediated model of neutrophilic asthma, we demonstrate that NRP2 was increased in AM and other lung antigen‐presenting cells following airway challenge with antigen. Conditional deletion of NRP2 in myeloid cells exacerbated airway inflammation in a neutrophilic asthma model. In contrast, myeloid‐specific ablation of NRP2 did not affect airway inflammation in a Th2‐mediated eosinophilic asthma model. Myeloid‐specific ablation of NRP2 did not affect Th1/Th17 responses to inhaled antigens or expression of neutrophil chemokines but rather resulted in impaired efferocytosis by AM, which is necessary for effective resolution of airway inflammation. Conclusion Our findings suggest that NRP2 is a negative regulator of airway inflammation associated with neutrophilic asthma.

Background:. High pressure injection injury to the hand with paint leads to amputation rates near 48%. Historically, authors utilized saline irrigation alone, but have high reoperation rates. We conducted a cadaveric study to determine the ideal detergent for effective paint removal from the soft tissue. Methods:. Two cadaveric hands were amputated from the same cadaver. The left and right hand digits were injected with flat white latex-based paint and flat white oil-based paint, respectively. Each digit received a longitudinal incision and was scrubbed for 120 seconds with 50 mL of a randomly assigned detergent and no detergent (saline) as the control. After achieving a lather, each finger was cleansed with 50 mL saline before being evaluated by two blinded hand surgery faculty. Reviewers assessed the washouts as adequate or inadequate, in order to generate a Kappa statistic and measure inter-rater reliability prior to ranking each digit (1 through 5) (ie, 1 = most paint-free soft tissue). Results:. The two hand faculty had an inter-rater reliability of 0.70. Both reviewers ranked povidone-iodine 10% or Johnson & Johnson shampoo as the best irrigant for latex-based paint. In oil-based paint, povidone-iodine 10%, Johnson & Johnson, & Techni-care were ranked as top three. All reviewers reported detergents were better than saline alone. Conclusions:. The addition of detergent created an irrigant that removed both latex- and oil-based paint better than normal saline alone. Based on these results, surgeons treating high-pressure injection injury should consider using Povidone-Iodine 10% or Johnson & Johnson baby shampoo for latex- or oil-based paint.

Sociologists have argued that there is value in incorporating computational tools into qualitative research, including using machine learning to code qualitative data. Yet standard computational approaches do not neatly align with traditional qualitative practices. The authors introduce a hybrid human-machine learning approach (HHMLA) that combines a contemporary iterative approach to qualitative coding with advanced word embedding models that allow contextual interpretation beyond what can be reliably accomplished with conventional computational approaches. The results, drawn from an analysis of 87 human-coded ethnographic interview transcripts, demonstrate that HHMLA can code data sets at a fraction of the effort of human-only strategies, saving hundreds of hours labor in even modestly sized qualitative studies, while improving coding reliability. The authors conclude that HHMLA may provide a promising model for coding data sets where human-only coding would be logistically prohibitive but conventional computational approaches would be inadequate given qualitative foci.

Ozone is a highly reactive environmental pollutant with well-recognized adverse effects on lung health. Bronchial hyperresponsiveness (BHR) is one consequence of ozone exposure, particularly for individuals with underlying lung disease. Our data demonstrated that ozone induced substantial ATP release from human airway epithelia in vitro and into the airways of mice in vivo and that ATP served as a potent inducer of mast cell degranulation and BHR, acting through P2X7 receptors on mast cells. Both mast cell–deficient and P2X7 receptor–deficient (P2X7–/–) mice demonstrated markedly attenuated BHR to ozone. Reconstitution of mast cell–deficient mice with WT mast cells and P2X7–/– mast cells restored ozone-induced BHR. Despite equal numbers of mast cells in reconstituted mouse lungs, mice reconstituted with P2X7–/– mast cells demonstrated significantly less robust BHR than mice reconstituted with WT mast cells. These results support a model where P2X7 on mast cells and other cell types contribute to ozone-induced BHR.

Abstract Background The efficient biological production of industrially and economically important compounds is a challenging problem. Brute-force determination of the optimal pathways to efficient production of a target chemical in a chassis organism is computationally intractable. Many current methods provide a single solution to this problem, but fail to provide all optimal pathways, optional sub-optimal solutions or hybrid biological/non-biological solutions. Results Here we present RetSynth, software with a novel algorithm for determining all optimal biological pathways given a starting biological chassis and target chemical. By dynamically selecting constraints, the number of potential pathways scales by the number of fully independent pathways and not by the number of overall reactions or size of the metabolic network. This feature allows all optimal pathways to be determined for a large number of chemicals and for a large corpus of potential chassis organisms. Additionally, this software contains other features including the ability to collect data from metabolic repositories, perform flux balance analysis, and to view optimal pathways identified by our algorithm using a built-in visualization module. This software also identifies sub-optimal pathways and allows incorporation of non-biological chemical reactions, which may be performed after metabolic production of precursor molecules. Conclusions The novel algorithm designed for RetSynth streamlines an arduous and complex process in metabolic engineering. Our stand-alone software allows the identification of candidate optimal and additional sub-optimal pathways, and provides the user with necessary ranking criteria such as target yield to decide which route to select for target production. Furthermore, the ability to incorporate non-biological reactions into the final steps allows determination of pathways to production for targets that cannot be solely produced biologically. With this comprehensive suite of features RetSynth exceeds any open-source software or webservice currently available for identifying optimal pathways for target production.

Abstract Introduction Sepsis results from a dysregulated host response to an infection that is associated with an imbalance between pro- and anti-inflammatory cytokines. This imbalance is hypothesized to be a driver of patient mortality. Certain autoimmune diseases modulate the expression of cytokines involved in the pathophysiology of sepsis. However, the outcomes of patients with autoimmune disease who develop sepsis have not been studied in detail. The objective of this study is to determine whether patients with autoimmune diseases have different sepsis outcomes than patients without these comorbidities. Methods Using the Multiparameter Intelligent Monitoring in Intensive Care III database (v. 1.4) which contains retrospective clinical data for over 50,000 adult ICU stays, we compared 30-day mortality risk for sepsis patients with and without autoimmune disease. We used logistic regression models to control for known confounders, including demographics, disease severity, and immunomodulation medications. We used mediation analysis to evaluate how the chronic use of immunomodulation medications affects the relationship between autoimmune disease and 30-day mortality. Results Our study found a statistically significant 27.00% reduction in the 30-day mortality risk associated with autoimmune disease presence. This association was found to be the strongest (OR 0.71, 95% CI 0.54–0.93, P = 0.014) among patients with septic shock. The autoimmune disease-30-day mortality association was not mediated through the chronic use of immunomodulation medications (indirect effect OR 1.07, 95% CI 1.01–1.13, P = 0.020). Conclusions We demonstrated that autoimmune diseases are associated with a lower 30-day mortality risk in sepsis. Our findings suggest that autoimmune diseases affect 30-day mortality through a mechanism unrelated to the chronic use of immunomodulation medications. Since this study was conducted within a single study center, research using data from other medical centers will provide further validation.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by accumulation of misfolded α-synuclein within the central nervous system (CNS). Visual problems in PD patients are common, although retinal pathology associated with PD is not well understood. The purpose of this study was to investigate retinal pathology in a transgenic mouse model (TgM83) expressing the human A53T α-synuclein mutation and assess the effect of α-synuclein “seeding” on the development of retinal pathology. Two-month-old TgM83 mice were intracerebrally inoculated with brain homogenate from old (12–18 months) TgM83 mice. Retinas were then analyzed at 5 months of age. We analyzed retinas from 5-month-old and 8-month-old uninoculated healthy TgM83 mice, and old (12–18 months) mice that were euthanized following the development of clinical signs. Retinas of B6C3H mice (genetic background of the TgM83 mouse) served as control. We used immunohistochemistry and western blot analysis to detect accumulation of α-synuclein, pTauThr231, inflammation, changes in macroautophagy, and cell death. Raman spectroscopy was used to test the potential to differentiate between retinal tissues of healthy mice and diseased mice. This work demonstrates retinal changes associated with the A53T mutation. Retinas of non-inoculated TgM83 mice had accumulation of α-synuclein, “pre-tangle” tau, activation of retinal glial cells, and photoreceptor cell loss by 8 months of age. The development of these changes is accelerated by inoculation with brain homogenate from clinically ill TgM83 mice. Compared to non-inoculated 5-month-old TgM83 mice, retinas of inoculated 5-month-old mice had increased accumulation of α-synuclein (pSer129) and pTauThr231 proteins, upregulated microglial activation, and dysregulated macroautophagy. Raman spectroscopic analysis was able to discriminate between healthy and diseased mice. This study describes retinal pathology resulting from the A53T mutation. We show that seeding with brain homogenates from old TgM83 mice accelerates retinal pathology. We demonstrate that Raman spectroscopy can be used to accurately identify a diseased retina based on its biochemical profile, and that α-synuclein accumulation may contribute to accumulation of pTauThr231 proteins, neuroinflammation, metabolic dysregulation, and photoreceptor cell death. Our work provides insight into retinal changes associated with Parkinson's disease, and may contribute to a better understanding of visual symptoms experienced by patients.

Cryptococcus neoformans

Neuroendocrine small cell carcinoma of the uterine cervix portends a dismal prognosis with limited treatment options. Rarely, tumors of mixed-lineage appear in gynecologic malignancies. Here, we report a 77-year-old woman who presented with complete uterine prolapse and 4-month history of vaginal bleeding. Histopathologic evaluation revealed a mixed adenoid cystic carcinoma and neuroendocrine small cell carcinoma of the uterine cervix. The tumor was PD-L1 and HPV 35 positive. The patient was treated with up-front surgery and adjuvant radiation. Independent, histology-specific alterations in FGFR2 and a FGFR2-TACC2 fusion were identified. Progression of disease occurred within 6 months for which she received chemotherapy and immunotherapy. However, the patient expired within a year. We comprehensively review how screening for and targeting of FGFR alterations in recurrent and metastatic cervical cancer might serve as a touchstone for future treatment regimens.

BackgroundPredictive models can serve as early warning systems and can be used to forecast future risk of various infectious diseases. Conventionally, regression and time series models are used to forecast dengue incidence, using dengue surveillance (e.g., case counts) and weather data. However, these models may be limited in terms of model assumptions and the number of predictors that can be included. Machine learning (ML) methods are designed to work with a large number of predictors and thus offer an appealing alternative. Here, we compared the performance of ML algorithms with that of regression models in predicting dengue cases and outbreaks from 4 to up to 12 weeks in advance. Many countries lack sufficient health surveillance infrastructure, as such we evaluated the contribution of dengue surveillance and weather data on the predictive power of these models.MethodsWe developed ML, regression, and time series models to forecast weekly dengue case counts and outbreaks in Iquitos, Peru; San Juan, Puerto Rico; and Singapore from 1990-2016. Forecasts were generated using available weekly dengue surveillance, and weather data. We evaluated the agreement between model forecasts and actual dengue observations using Mean Absolute Error and Matthew's Correlation Coefficient (MCC).ResultsFor near term predictions of weekly case counts and when using surveillance data, ML models had 21% and 33% less error than regression and time series models respectively. However, using weather data only, ML models did not demonstrate a practical advantage. When forecasting weekly dengue outbreaks 12 weeks in advance, ML models achieved a maximum MCC of 0.61.ConclusionsOur results identified 2 scenarios when ML models are advantageous over regression model: 1) predicting dengue weekly case counts 4 weeks ahead when dengue surveillance data are available and 2) predicting weekly dengue outbreaks 12 weeks ahead when dengue surveillance data are unavailable. Given the advantages of ML models, dengue early warning systems may be improved by the inclusion of these models.

Summary: The mammary epithelial cell (MEC) system is a bilayered ductal epithelium of luminal and basal cells, maintained by a lineage of stem and progenitor populations. Here, we used integrated single-cell transcriptomics and chromatin accessibility analysis to reconstruct the cell types of the mouse MEC system and their underlying gene regulatory features in an unbiased manner. We define differentiation states within the secretory type of luminal cells, which forms a continuous spectrum of general luminal progenitor and lactation-committed progenitor cells. By integrating single-cell transcriptomics and chromatin accessibility landscapes, we identify cis- and trans-regulatory elements that are differentially activated in the specific epithelial cell types and our newly defined luminal differentiation states. Our work provides a resource to reveal cis/trans-regulatory elements associated with MEC identity and differentiation that will serve as a reference to determine how the chromatin accessibility landscape changes during breast cancer.

Cerebral Blood Flow Velocity waveforms acquired via Transcranial Doppler (TCD) can provide evidence for cerebrovascular occlusion and stenosis. Thrombolysis in Brain Ischemia (TIBI) flow grades are widely used for this purpose, but require subjective assessment by expert evaluators to be reliable. In this work we seek to determine whether TCD morphology can be objectively assessed using an unsupervised machine learning approach to waveform categorization. TCD beat waveforms were recorded at multiple depths from the Middle Cerebral Arteries of 106 subjects; 33 with Large Vessel Occlusion (LVO). From each waveform, three morphological features were extracted, quantifying onset of maximal velocity, systolic canopy length, and the number/prominence of peaks/troughs. Spectral clustering identified groups implicit in the resultant three-dimensional feature space, with gap statistic criteria establishing the optimal cluster number. We found that gap statistic disparity was maximized at four clusters, referred to as flow types I, II, III, and IV. Types I and II were primarily composed of control subject waveforms, whereas types III and IV derived mainly from LVO patients. Cluster morphologies for types I and IV aligned clearly with Normal and Blunted TIBI flows, respectively. Types II and III represented commonly observed flow-types not delineated by TIBI, which nonetheless deviate from normal and blunted flows. We conclude that important morphological variability exists beyond that currently quantified by TIBI in populations experiencing or at-risk for acute ischemic stroke, and posit that the observed flow-types provide the foundation for objective methods of real-time automated flow type classification.

Autophagy is a major intracellular degradation process that is essential for the clearance of unnecessary proteins/organelles and the maintenance of cellular homeostasis. The inhibition of autophagy results in cellular consequences associated with many skeletal muscle pathologies, and therapies designed to elevate autophagic activity may provide protection from such pathologies. Acute exposure to low levels of heat has therapeutic effects; however, the impact of heat on skeletal muscle autophagy remains unclear. In the present study, C2C12 myotubes were maintained at 37°C thermoneutral (TN) or heated at 40°C heat treatment (HT) for 1 h. Myotubes were harvested immediately after heating, or returned to 37°C for recovery of 2 or 24 h. HT resulted in an elevation in pAMPK (T172), Beclin-1, and LC3 II, a marker for autophagosome formation, but no change in p62. In the context of autophagy inhibition with Bafilomycin A1, HT resulted in lower LC3 II compared to TN. The applied heat load induced the heat shock response, as evidenced by immediate upregulation of HSF1 and Hsp70. Hsp70 continued to increase during recovery, whereas pHsp27 was downregulated acutely in response to HT, but retuned to TN levels by 2 h of recovery. HT also reduced the phosphorylation of the MAP-kinases p38 and JNK. These findings suggest that an acute, short bout of mild heat may be beneficial to skeletal muscle by increasing AMPK activity, markers of autophagasome formation, and the heat shock response.

In a no-till system, there are many different methods available for terminating cover crops. Mechanical termination, utilizing rolling and crimping technology, is one method that injures the plant without cutting the stems. Another popular and commercially available method is mowing, but this can cause problems with cover crop re-growth and loose residue interfering with the planter during cash crop planting. A field experiment was conducted over three growing seasons in northern Alabama to determine the effects of different cover crops and termination methods on cantaloupe yield in a no-till system. Crimson clover, cereal rye, and hairy vetch cover crops were terminated using two different roller-crimpers, including a two-stage roller-crimper for four-wheel tractors and a powered roller-crimper for a two-wheel walk-behind tractor. Cover crop termination rates were evaluated one, two, and three weeks after termination. Three weeks after rolling, a higher termination rate was found for flail mowing (92%) compared to lower termination rates for a two-stage roller (86%) and powered roller-crimper (85%), while the control termination rate was only 49%. There were no significant differences in cantaloupe yield among the rolling treatments, which averaged 38,666 kg ha−1. However, yields were higher for cereal rye and hairy vetch cover crops (41,785 kg ha−1 and 42,000 kg ha−1) compared to crimson clover (32,213 kg ha−1).

Blastomycosis is a rare fungal infection that frequently involves the skin. Atypical presentations are important to identify, especially in pregnant patients, to initiate appropriate therapy and prevent complications. Uniquely, we describe a case of atypical blastomycosis that presented with painful cutaneous abscesses in a pregnant patient, with dissemination to the central nervous system. The case was successfully treated with liposomal amphotericin B transitioned to voriconazole after delivery without complications for the patient or fetus. Keywords: Blastomycosis, pregnancy, amphotericin B

Abstract Glioneuronal tumors constitute a histologically diverse group of primary central nervous system neoplasms that are typically slow-growing and managed conservatively. Genetic alterations associated with glioneuronal tumors include BRAF mutations and oncogenic fusions. To further characterize this group of tumors, we collected a cohort of 26 glioneuronal tumors and performed in-depth genomic analysis. We identified mutations in BRAF (34%) and oncogenic fusions (30%), consistent with previously published reports. In addition, we discovered novel oncogenic fusions involving members of the NTRK gene family in a subset of our cohort. One-patient with BCAN exon 13 fused to NTRK1 exon 11 initially underwent a subtotal resection for a 4th ventricular glioneuronal tumor but ultimately required additional therapy due to progressive, symptomatic disease. Given the patient’s targetable fusion, the patient was enrolled on a clinical trial with entrectinib, a pan-Trk, ROS1, and ALK (anaplastic lymphoma kinase) inhibitor. The patient was treated for 11 months and during this time volumetric analysis of the lesion demonstrated a maximum reduction of 60% in the contrast-enhancing tumor compared to his pre-treatment magnetic resonance imaging study. The radiologic response was associated with resolution of his clinical symptoms and was maintained for 11 months on treatment. This report of a BCAN-NTRK1 fusion in glioneuronal tumors highlights its clinical importance as a novel, targetable alteration.