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Objective: Inpatient vaccination is an opportunity to increase vaccine uptake among patients at high risk for severe COVID-19 illness. We designed and implemented a hospital-based COVID-19 vaccination program with the aim of increasing documentation of vaccine eligibility and COVID-19 vaccination to eligible inpatients before discharge., Methods: We integrated a templated note into the electronic medical records and trained health care personnel to screen inpatients and document COVID-19 vaccine eligibility at the Atlanta Veterans Affairs Medical Center. Vaccination staff deployed to inpatient wards administered the vaccine to eligible and consenting patients at the bedside. We calculated the number of inpatients whose vaccine eligibility was assessed and documented during a 4-week period after health care personnel were trained. We used the Wald χ 2 test to compare the proportion of eligible patients who were vaccinated before discharge 4 weeks before (March 29-April 23, 2021) and 4 weeks after (May 3-28, 2021) the training period., Results: During the 4 weeks before the training period, COVID-19 vaccine eligibility was not routinely assessed and documented. Of 793 inpatients discharged during the 4 weeks after the training period, 470 (59%) had COVID-19 vaccine eligibility documented. Of 86 patients who were eligible for vaccination, 61 (71%) received COVID-19 vaccination before discharge. COVID-19 vaccination rates during hospitalization increased significantly from 16 of 769 inpatients (2%) during the 4 weeks before training to 61 of 793 inpatients (8%) during the 4 weeks after training ( P  < .001)., Conclusion: An inpatient vaccination program that integrated COVID-19 vaccination into discharge planning increased vaccine screening and uptake. Future studies are needed to identify barriers to vaccination and strategies to increase vaccine uptake among those who are hesitant., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Poor air quality is linked to numerous adverse health effects including strokes, heart attacks, and premature death. Improving energy efficiency in the industrial sector reduces air emissions and yields health benefits. One of these strategies, replacing an existing grid boiler (GB) with a combined heat and power (CHP) system, can improve a facility's energy efficiency but can also increase local air emissions, which in turn can affect health outcomes. Previous studies have considered air-emissions and health outcomes of CHP system installation at a single location, but few studies have investigated the regional air quality and health impacts of replacing an existing GB with new CHP system. This study estimates the emission changes and associated health impacts of this shift in 14 regions in the US, representing different electricity generation profiles. It assumes that one manufacturing facility in each region switches from an existing GB to a CHP system. The monetized annual US health benefits of shifting a single GB to a CHP in each of the 14 regions range from $-5.3 to 0.55 million (2022 USD), while including CHP emission control increases the benefits by 100-170% ($9,000 to 1.15 million (2022 USD)). This study also includes a sensitivity analysis, which suggests that the facility location (region, state, and county), boiler efficiency, and emission control of the CHP are key factors that would determine whether shifting from a GB to CHP system would result in health benefits or burdens. Implications : Combined heat and power (CHP) systems offer industrial facilities the opportunity to improve their energy efficiency and reduce greenhouse gas emissions. However, CHP systems also combust more fuel on site and can also increase local air emissions. This study evaluates how converting an existing grid boiler (GB) system to a CHP system (with or without emission control) affects local (from combustion) and regional emissions (from electricity consumption) and the associated health burdens in different US regions. A facility can use this study's analysis as an example for estimating the tradeoffs between local emission changes, regional emission changes, and health effects. It also provides a comparison between the incremental cost of adding SCR (compared to uncontrolled CHPs) and the NPV of the monetized health benefits associated with adding the SCR.

Context: Between April 2020 and May 2021, the Centers for Disease Control and Prevention (CDC) awarded more than $40 billion to health departments nationwide for COVID-19 prevention and response activities. One of the identified priorities for this investment was improving infection prevention and control (IPC) in nursing homes., Program: CDC developed a virtual course to train new and less experienced public health staff in core healthcare IPC principles and in the application of CDC COVID-19 healthcare IPC guidance for nursing homes., Implementation: From October 2020 to August 2021, the CDC led training sessions for 12 cohorts of public health staff using pretraining reading materials, case-based scenarios, didactic presentations, peer-learning opportunities, and subject matter expert-led discussions. Multiple electronic assessments were distributed to learners over time to measure changes in self-reported knowledge and confidence and to collect feedback on the course. Participating public health programs were also assessed to measure overall course impact., Evaluation: Among 182 enrolled learners, 94% completed the training. Most learners were infection preventionists (42%) or epidemiologists (38%), had less than 1 year of experience in their health department role (75%), and had less than 1 year of subject matter experience (54%). After training, learners reported increased knowledge and confidence in applying the CDC COVID-19 healthcare IPC guidance for nursing homes (≥81%) with the greatest increase in performing COVID-19 IPC consultations and assessments (87%). The majority of participating programs agreed that the course provided an overall benefit (88%) and reduced training burden (72%)., Discussion: The CDC's virtual course was effective in increasing public health capacity for COVID-19 healthcare IPC in nursing homes and provides a possible model to increase IPC capacity for other infectious diseases and other healthcare settings. Future virtual healthcare IPC courses could be enhanced by tailoring materials to health department needs, reinforcing training through applied learning experiences, and supporting mechanisms to retain trained staff., Competing Interests: The authors declare no conflicts of interest.

Objective: This study describes characteristics of large tuberculosis (TB) outbreaks in the United States detected using novel molecular surveillance methods during 2014-2016 and followed for 2 years through 2018., Methods: We developed 4 genotype-based detection algorithms to identify large TB outbreaks of ≥10 cases related by recent transmission during a 3-year period. We used whole-genome sequencing and epidemiologic data to assess evidence of recent transmission among cases., Results: There were 24 large outbreaks involving 518 cases; patients were primarily U.S.-born (85.1%) racial/ethnic minorities (84.1%). Compared with all other TB patients, patients associated with large outbreaks were more likely to report substance use, homelessness, and having been diagnosed while incarcerated. Most large outbreaks primarily occurred within residences among families and nonfamilial social contacts. A source case with a prolonged infectious period and difficulties in eliciting contacts were commonly reported contributors to transmission., Conclusion: Large outbreak surveillance can inform targeted interventions to decrease outbreak-associated TB morbidity., Competing Interests: Declaration of competing interest None., (Published by Elsevier Ltd.)

The purpose of this study is to examine forensic documentation of non-fatal strangulation (NFS) in domestic violence cases. Research has pointed to the importance of forensic evidence in the prosecution of strangulation offenders. However, limited research has examined the type of evidence that is gathered during a forensic examination for NFS that occurs during a domestic violence situation. To address this gap in the literature, this study analyzed 63 NFS forensic examination records and body sketches for victims who were referred by police during a domestic violence incident. Results reflect the seriousness of NFS with multiple strangulation attacks in the current incident recorded in 52% of the reports, loss of consciousness recorded in 13.1% of the reports, and a history of strangulation in the relationship recorded in 60% of the reports. It is argued that a forensic exam is essential to identify and mitigate serious symptoms and injuries, as well as to collect valuable evidence that can be used during legal proceedings. Implications for practice and policy are discussed., (© 2022 American Academy of Forensic Sciences.)

Purpose: Raloxifene (RAL) is a selective estrogen receptor modulator (SERM) that has previously been shown to cause acellular benefits to bone tissue. Due to these improvements, RAL was combined with targeted tibial loading to assess if RAL treatment during periods of active bone formation would allow for further mechanical enhancements. Methods: Structural, mechanical, and microstructural effects were assessed in bone from C57BL/6 mice that were treated with RAL (0.5 mg/kg), tibial loading, or both for 6 weeks, beginning at 10 weeks of age. Results: Ex vivo microcomputed tomography (CT) images indicated RAL and loading work together to improve bone mass and architecture, especially within the cancellous region of males. Increases in cancellous bone volume fraction were heavily driven by increases in trabecular thickness, though there were some effects on trabecular spacing and number. In the cortical regions, RAL and loading both increased cross-sectional area, cortical area, and cortical thickness. Whole-bone mechanical testing primarily indicated the effects of loading. Further characterization through Raman spectroscopy and nanoindentation showed load-based changes in mineralization and micromechanics, while both loading and RAL caused changes in the secondary collagen structure. In contrast to males, in females, there were large load-based effects in the cancellous and cortical regions, resulting in increased whole-bone mechanical properties. RAL had less of an effect on cancellous and cortical architecture, though some effects were still present. Conclusion:  RAL and loading work together to impact bone architecture and mechanical integrity, leading to greater improvements than either treatment individually.

To meet targets for reducing greenhouse gas emissions, many countries, including Estonia, must transition to low-emission electricity sources. Based on current circumstances, the most likely options in Estonia are renewables with energy storage, oil shale power plants with carbon capture and storage (CCS), or the combination of renewables and either oil shale or nuclear power plants. Here we compare these different scenarios to help determine which would be the most promising based on current information. For the comparison we performed simulations to assess how various systems meet the electricity demand in Estonia and at what cost. Based on our simulation results and literature data, combining wind turbines with thermal power plants would provide grid stability at a more affordable cost. Using nuclear power to compliment wind turbines would lead to an overall levelized cost of electricity (LCOE) in the range of 68 to 150 EUR/MWh (median of 103 EUR/MWh). Using oil shale power plants with CCS would give a cost between 91 and 163 EUR/MWh (median of 118 EUR/MWh). By comparison, using only renewables and energy storage would have an LCOE of 106 to 241 EUR/MWh (median of 153 EUR/MWh)., Competing Interests: The authors have declared that no competing interests exist.

Introduction: Survey results from 2016 and 2018 at the University of Michigan College of Pharmacy highlighted mental health concerns for the student population, including struggles with depression, anxiety, and academic distress. This led to creation of a pilot well-being elective course for first year doctor of pharmacy students. This article describes how this course was assessed and adapted for the future., Methods: The well-being elective course used a course-specific survey and the Brief Inventory of Thriving to assess student outcomes. The course-specific survey was based upon the course objectives and the University of Michigan Common Agenda for Well-Being. Both surveys were given pre- and post-course to identify change., Results: Course survey results illustrated an improvement in student well-being over a single semester. Compared with pre-course responses, students who completed the course were significantly more likely to agree with statements indicating they had strong time management skills, resilience to manage the fluctuations of life, were able to make thoughtful choices to reduce harm and promote well-being, and overall rated their well-being as excellent. Additionally, nearly all students felt a sense of strengthened community with peers and faculty within the course, better able to recognize or refer a peer for help, and felt the course contributed to their overall well-being., Conclusions: Implementation of this well-being elective pilot course provided students the tools and resources to improve upon their overall well-being in an effort to address anxiety, depression, and academic distress., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Importance: The number of clinics marketing stem cell products for joint diseases, chronic pain, and most recently, COVID-19, has increased despite warnings from the US Food and Drug Administration that stem cell products for these and other indications have not been proven safe or effective., Objective: To examine bacterial infections in 20 patients who received umbilical cord blood-derived products marketed as stem cell treatment., Design, Setting, and Participants: This case series is a national public health investigation including case-finding, medical record review and abstraction, and laboratory investigation, including sterility testing of products and whole-genome sequencing of patient and product isolates. Participants included patients who developed bacterial infections following administration of umbilical cord blood-derived products marketed as stem cell treatment during August 2017 to September 2018. Data analysis was performed from March 2019 to September 2021., Exposures: Umbilical cord blood-derived products marketed as stem cell treatment., Main Outcomes and Measures: Data were collected on patient infections and exposures. The Centers for Disease Control and Prevention performed sterility testing on undistributed and distributed vials of product marketed as stem cell treatment and performed whole-genome sequencing to compare patient and product bacterial isolates., Results: Culture-confirmed bacterial infections were identified in 20 patients (median [range] age, 63 [2-89] years; 13 male patients [65%]) from 8 US states who sought stem cell treatment for conditions including pain, osteoarthritis, rheumatoid arthritis, and injury; all but 1 required hospitalization. The most frequently isolated bacteria from patients with infections were common enteric species, including Escherichia coli (14 patients) and Enterobacter cloacae (7 patients). Of unopened, undistributed products sampled for testing, 65% (22 of 34 vials) were contaminated with at least 1 of 16 bacterial species, mostly enteric. A patient isolate from Arizona matched isolates obtained from products administered to patients in Florida, and patient isolates from Texas matched undistributed product sent from the company in California., Conclusions and Relevance: Unapproved stem cell products can expose patients to serious risks without proven benefit. Sequencing results suggest a common source of extensive contamination, likely occurring during the processing of cord blood into product. Patients and health care practitioners who are considering the use of unapproved products marketed as stem cell treatment should be aware of their unproven benefits and potential risks, including serious infections.

While research tends to focus on understanding risk factors associated with adverse childhood and adolescent experiences, there is growing empirical support within the field of human services to also focus on strengths and protective factors that bolster resilience in life. This study investigated the psychometrics of the Resilience Protective Factors Checklist (RPFC), which is a questionnaire that assists in the identification of protective factors empirically linked to resilience and positive outcomes. In addition, associations between protective factors, childhood adversity, and quality of life outcomes were examined. Participants were undergraduate college students ( n  = 1,256). The results revealed good psychometric properties for the RPFC and the factor analysis supported the resilience structural model of three interrelated areas of protective factors - Individual, Family, and Community influences. All three areas of protection were positively correlated with all four domains of positive life outcomes (physical health, psychological health, social relationships, and a healthy environment) and negatively correlated with adverse childhood experiences. RPFC's individual protective factors positively predicted physical health and psychological health outcomes, while the community protective factors positively predicted environmental health outcomes. In addition, the RPFC's family protective factors moderated the path between adverse childhood experiences and social relationships. That is, for individuals who have experienced more adverse childhood experiences, those with higher levels of RPFC family protective factors had positive social relationship outcomes, while those with lower levels of family protective factors had negative social relationship outcomes. These findings lend support for the utilization of assessment measures for identifying and promoting protective factors that can buffer life adversity and heighten resilient positive outcomes.

Introduction: In the U.S., universal genotyping of culture-confirmed tuberculosis cases facilitates cluster detection. Early recognition of the small clusters more likely to become outbreaks can help prioritize public health resources for immediate interventions., Methods: This study used national surveillance data reported during 2009-2018 to describe incident clusters (≥3 tuberculosis cases with matching genotypes not previously reported in the same county); data were analyzed during 2020. Cox proportional hazards regression models were used to examine the patient characteristics associated with clusters doubling in size to ≥6 cases., Results: During 2009-2018, a total of 1,516 incident clusters (comprising 6,577 cases) occurred in 47 U.S. states; 231 clusters had ≥6 cases. Clusters of ≥6 cases disproportionately included patients who used substances, who had recently experienced homelessness, who were incarcerated, who were U.S. born, or who self-identified as being of American Indian or Alaska Native race or of Black race. A median of 54 months elapsed between the first and the third cases in clusters that remained at 3-5 cases compared with a median of 9.5 months in clusters that grew to ≥6 cases. The longer time between the first and third cases and the presence of ≥1 patient aged ≥65 years among the first 3 cases predicted a lower hazard for accumulating ≥6 cases., Conclusions: Clusters accumulating ≥3 cases within a year should be prioritized for intervention. Effective response strategies should include plans for targeted outreach to U.S.-born individuals, incarcerated people, those experiencing homelessness, people using substances, and individuals self-identifying as being of American Indian or Alaska Native race or of Black race., (Published by Elsevier Inc.)

MYCN is amplified in 20% to 25% of neuroblastoma, and MYCN -amplified neuroblastoma contributes to a large percent of pediatric cancer-related deaths. Therapy improvements for this subtype of cancer are a high priority. Here we uncover a MYCN-dependent therapeutic vulnerability in neuroblastoma. Namely, amplified MYCN rewires the cell through expression of key receptors, ultimately enhancing iron influx through increased expression of the iron import transferrin receptor 1. Accumulating iron causes reactive oxygen species (ROS) production, and MYCN -amplified neuroblastomas show enhanced reliance on the system Xc- cystine/glutamate antiporter for ROS detoxification through increased transcription of this receptor. This dependence creates a marked vulnerability to targeting the system Xc-/glutathione (GSH) pathway with ferroptosis inducers. This reliance can be exploited through therapy with FDA-approved rheumatoid arthritis drugs sulfasalazine (SAS) and auranofin: in MYCN -amplified, patient-derived xenograft models, both therapies blocked growth and induced ferroptosis. SAS and auranofin activity was largely mitigated by the ferroptosis inhibitor ferrostatin-1, antioxidants like N-acetyl-L-cysteine, or by the iron scavenger deferoxamine (DFO). DFO reduced auranofin-induced ROS, further linking increased iron capture in MYCN -amplified neuroblastoma to a therapeutic vulnerability to ROS-inducing drugs. These data uncover an oncogene vulnerability to ferroptosis caused by increased iron accumulation and subsequent reliance on the system Xc-/GSH pathway. SIGNIFICANCE: This study shows how MYCN increases intracellular iron levels and subsequent GSH pathway activity and demonstrates the antitumor activity of FDA-approved SAS and auranofin in patient-derived xenograft models of MYCN -amplified neuroblastoma., (©2021 American Association for Cancer Research.)

We analyzed data from 2012 to 2016 for patients who were hospitalized or who died after ≥1 dose of isoniazid-rifapentine for treatment of latent Mycobacterium tuberculosis infection. No patients died; 15 were hospitalized. Nine patients experienced hypotension, and 5 had elevated serum aminotransferases, reinforcing the need for vigilant monitoring during treatment., (© Published by Oxford University Press for the Infectious Diseases Society of America 2020.)

Ozone (O 3 ) is a potent oxidant associated with adverse health effects. Low-cost O 3 sensors, such as metal oxide (MO) sensors, can complement regulatory O 3 measurements and enhance the spatiotemporal resolution of measurements. However, the quality of MO sensor data remains a challenge. The University of Utah has a network of low-cost air quality sensors (called AirU) that primarily measures PM 2.5 concentrations around the Salt Lake City valley (Utah, U.S.). The AirU package also contains a low-cost MO sensor ($8) that measures oxidizing/reducing species. These MO sensors exhibited excellent laboratory response to O 3 although they exhibited some intra-sensor variability. Field performance was evaluated by placing eight AirUs at two Division of Air Quality (DAQ) monitoring stations with O 3 federal equivalence methods for one year to develop long-term multiple linear regression (MLR) and artificial neural network (ANN) calibration models to predict O 3 concentrations. Six sensors served as train/test sets. The remaining two sensors served as a holdout set to evaluate the applicability of the new calibration models in predicting O 3 concentrations for other sensors of the same type. A rigorous variable selection method was also performed by least absolute shrinkage and selection operator (LASSO), MLR and ANN models. The variable selection indicated that the AirU's MO oxidizing species and temperature measurements and DAQ's solar radiation measurements were the most important variables. The MLR calibration model exhibited moderate performance (R 2  = 0.491), and the ANN exhibited good performance (R 2  = 0.767) for the holdout set. We also evaluated the performance of the MLR and ANN models in predicting O 3 for five months after the calibration period and the results showed moderate correlations (R 2 s of 0.427 and 0.567, respectively). These low-cost MO sensors combined with a long-term ANN calibration model can complement reference measurements to understand geospatial and temporal differences in O 3 levels., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Drs. Gaillardon and Kelly, co-authors on this paper, have a financial interest in the company Tetrad: Sensor Network Solutions, LCC, which commercializes solutions for environmental monitoring. Several authors of this paper (Sayahi, Butterfield, Quah, Garff, Becnel, Le, Gaillardon, Kelly) have submitted an invention disclosure related to this paper to the University of Utah technology venture and commercialization office., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Exercise benefits the musculoskeletal system and reduces the effects of cancer. The effects of exercise are multifactorial, where metabolic changes and tissue adaptation influence outcomes. Mechanical signals, a principal component of exercise, are anabolic to the musculoskeletal system and restrict cancer progression. We examined the mechanisms through which cancer cells sense and respond to low-magnitude mechanical signals introduced in the form of vibration. Low-magnitude, high-frequency vibration was applied to human breast cancer cells in the form of low-intensity vibration (LIV). LIV decreased matrix invasion and impaired secretion of osteolytic factors PTHLH, IL-11, and RANKL. Furthermore, paracrine signals from mechanically stimulated cancer cells, reduced osteoclast differentiation and resorptive capacity. Disconnecting the nucleus by knockdown of SUN1 and SUN2 impaired LIV-mediated suppression of invasion and osteolytic factor secretion. LIV increased cell stiffness; an effect dependent on the LINC complex. These data show that mechanical vibration reduces the metastatic potential of human breast cancer cells, where the nucleus serves as a mechanosensory apparatus to alter cell structure and intercellular signaling.

Recent advances in fabrication have enabled radial-junction architectures for cost-effective and high-performance optoelectronic devices. Unlike a planar PN junction, a radial-junction geometry maximizes the optical interaction in the three-dimensional (3D) structures, while effectively extracting the generated carriers via the conformal PN junction. In this paper, we report characterizations of radial PN junctions that consist of p-type Si micropillars created by deep reactive-ion etching (DRIE) and an n-type layer formed by phosphorus gas diffusion. We use electron-beam induced current (EBIC) microscopy to access the 3D junction profile from the sidewall of the pillars. Our EBIC images reveal uniform PN junctions conformally constructed on the 3D pillar array. Based on Monte-Carlo simulations and EBIC modeling, we estimate local carrier separation/collection efficiency that reflects the quality of the PN junction. We find the EBIC efficiency of the pillar array increases with the incident electron beam energy, consistent with the EBIC behaviors observed in a high-quality planar PN junction. The magnitude of the EBIC efficiency of our pillar array is about 70% at 10 kV, slightly lower than that of the planar device (≈ 81%). We suggest that this reduction could be attributed to the unpassivated pillar surface and the unintended recombination centers in the pillar cores introduced during the DRIE processes. Our results support that the depth-dependent EBIC approach is ideally suitable for evaluating PN junctions formed on micro/nanostructured semiconductors with various geometry.

Raloxifene (RAL) is an FDA-approved drug used to treat osteoporosis in postmenopausal women. RAL suppresses bone loss primarily through its role as a selective estrogen receptor modulator (SERM). This hormonal estrogen therapy promotes unintended side effects, such as hot flashes and increased thrombosis risk, and prevents the drug from being used in some patient populations at-risk for fracture, including children with bone disorders. It has recently been demonstrated that RAL can have significant positive effects on overall bone mechanical properties by binding to collagen and increasing bone tissue hydration in a cell-independent manner. A Raloxifene-Analog (RAL-A) was synthesized by replacing the 6-hydroxyl substituent with 6-methoxy in effort to reduce the compound's binding affinity for estrogen receptors (ER) while maintaining its collagen-binding ability. It was hypothesized that RAL-A would improve the mechanical integrity of bone in a manner similar to RAL, but with reduced estrogen receptor binding. Molecular assessment showed that while RAL-A did reduce ER binding, downstream ER signaling was not completely abolished. In-vitro , RAL-A performed similarly to RAL and had an identical concentration threshold on osteocyte cell proliferation, differentiation, and function. To assess treatment effect in-vivo , wildtype (WT) and heterozygous (OIM+/-) female mice from the Osteogenesis Imperfecta (OI) murine model were treated with either RAL or RAL-A from 8 weeks to 16 weeks of age. There was an untreated control group for each genotype as well. Bone microarchitecture was assessed using microCT, and mechanical behavior was assessed using 3-point bending. Results indicate that both compounds produced analogous gains in tibial trabecular and cortical microarchitecture. While WT mechanical properties were not drastically altered with either treatment, OIM+/- mechanical properties were significantly enhanced, most notably, in post-yield properties including bone toughness. This proof-of-concept study shows promising results and warrants the exploration of additional analog iterations to further reduce ER binding and improve fracture resistance., Competing Interests: There are no known conflicts of interest associated with this publication, and there has been no financial support for this work that could have influenced its outcome., (© 2020 The Authors.)

Competing Interests: All authors have completed and submitted the International Committee of Medical Journal Editors form for disclosure of potential conflicts of interest. No potential conflicts of interest were disclosed.

Current interventions to reduce skeletal fragility are insufficient at enhancing both the quantity and quality of bone when attempting to improve overall mechanical integrity. Bisphosphonates, such as Zoledronate (ZOL), are used to treat a variety of bone disorders by increasing bone mass to decrease fracture risk, but long-term use has been shown in some settings to compromise bone quality. Alternatively, Raloxifene (RAL) has recently been demonstrated to improve tissue quality and overall mechanical properties in a cell-independent manner by binding to collagen and increasing tissue hydration. We hypothesized that a combination of RAL and ZOL would improve mechanical and material properties of bone more than either monotherapy alone by enhancing both quantity and quality. In this study, wildtype (WT) and heterozygous (OIM+/-) male mice from the Osteogenesis Imperfecta (OI) murine model were treated with either RAL, ZOL, or both from 8 weeks to 16 weeks of age. Using the OIM model allows for investigation of therapeutic effects on a quality-based bone disease. Combination treatment resulted in higher trabecular architecture, cortical mechanical properties, and cortical fracture toughness in diseased mouse bone. Two fracture toughness properties, which are direct measures of the tissue's ability to resist the initiation and propagation of a crack, were significantly improved with combination treatment in OIM+/- compared to control. There was no significant effect on fracture toughness with either monotherapy alone in either genotype. Following the mass-based effects of ZOL, trabecular bone volume fraction was significantly higher with combination treatment in both genotypes. Combination treatment resulted in higher ultimate stress in both genotypes. RAL and combination treatment in OIM+/- also increased resilience compared to the control. In conclusion, this study demonstrates the beneficial effects of using combination drug treatments to increase bone mass while simultaneously improving tissue quality, especially to enhance the mechanical integrity of diseased bone. Combination therapies could be a potential method to improve bone health and combat skeletal fragility on both the microscopic and macroscopic levels., (Copyright © 2019 Elsevier Inc. All rights reserved.)

On September 6, 2019, this report was posted as an MMWR Early Release on the MMWR website (https://www.cdc.gov/mmwr). As of August 27, 2019, 215 possible cases of severe pulmonary disease associated with the use of electronic cigarette (e-cigarette) products (e.g., devices, liquids, refill pods, and cartridges) had been reported to CDC by 25 state health departments. E-cigarettes are devices that produce an aerosol by heating a liquid containing various chemicals, including nicotine, flavorings, and other additives (e.g., propellants, solvents, and oils). Users inhale the aerosol, including any additives, into their lungs. Aerosols produced by e-cigarettes can contain harmful or potentially harmful substances, including heavy metals such as lead, volatile organic compounds, ultrafine particles, cancer-causing chemicals, or other agents such as chemicals used for cleaning the device (1). E-cigarettes also can be used to deliver tetrahydrocannabinol (THC), the principal psychoactive component of cannabis, or other drugs; for example, "dabbing" involves superheating substances that contain high concentrations of THC and other plant compounds (e.g., cannabidiol) with the intent of inhaling the aerosol. E-cigarette users could potentially add other substances to the devices. This report summarizes available information and provides interim case definitions and guidance for reporting possible cases of severe pulmonary disease. The guidance in this report reflects data available as of September 6, 2019; guidance will be updated as additional information becomes available., Competing Interests: All authors have completed and submitted the International Committee of Medical Journal Editors form for disclosure of potential conflicts of interest. No potential conflicts of interest were disclosed.

Purpose: It was recently demonstrated that the EWSR1-FLI1 t(11;22)(q24;12) translocation contributes to the hypersensitivity of Ewing sarcoma to PARP inhibitors, prompting clinical evaluation of olaparib in a cohort of heavily pretreated Ewing sarcoma tumors. Unfortunately, olaparib activity was disappointing, suggesting an underappreciated resistance mechanism to PARP inhibition in patients with Ewing sarcoma. We sought to elucidate the resistance factors to PARP inhibitor therapy in Ewing sarcoma and identify a rational drug combination capable of rescuing PARP inhibitor activity., Experimental Design: We employed a pair of cell lines derived from the same patient with Ewing sarcoma prior to and following chemotherapy, a panel of Ewing sarcoma cell lines, and several patient-derived xenograft (PDX) and cell line xenograft models., Results: We found olaparib sensitivity was diminished following chemotherapy. The matched cell line pair revealed increased expression of the antiapoptotic protein BCL-2 in the chemotherapy-resistant cells, conferring apoptotic resistance to olaparib. Resistance to olaparib was maintained in this chemotherapy-resistant model in vivo , whereas the addition of the BCL-2/X L inhibitor navitoclax led to tumor growth inhibition. In 2 PDXs, olaparib and navitoclax were minimally effective as monotherapy, yet induced dramatic tumor growth inhibition when dosed in combination. We found that EWS-FLI1 increases BCL-2 expression; however, inhibition of BCL-2 alone by venetoclax is insufficient to sensitize Ewing sarcoma cells to olaparib, revealing a dual necessity for BCL-2 and BCL-X L in Ewing sarcoma survival., Conclusions: These data reveal BCL-2 and BCL-X L act together to drive olaparib resistance in Ewing sarcoma and reveal a novel, rational combination therapy that may be put forward for clinical trial testing., (©2018 American Association for Cancer Research.)

Competing Interests: All authors have completed and submitted the ICMJE form for disclosure of potential conflicts of interest. No potential conflicts of interest were disclosed.

Purpose: EGFR inhibitors (EGFRi) are effective against EGFR -mutant lung cancers. The efficacy of these drugs, however, is mitigated by the outgrowth of resistant cells, most often driven by a secondary acquired mutation in EGFR, T790M We recently demonstrated that T790M can arise de novo during treatment; it follows that one potential therapeutic strategy to thwart resistance would be identifying and eliminating these cells [referred to as drug-tolerant cells (DTC)] prior to acquiring secondary mutations like T790M Experimental Design: We have developed DTCs to EGFRi in EGFR -mutant lung cancer cell lines. Subsequent analyses of DTCs included RNA-seq, high-content microscopy, and protein translational assays. Based on these results, we tested the ability of MCL-1 BH3 mimetics to combine with EGFR inhibitors to eliminate DTCs and shrink EGFR -mutant lung cancer tumors in vivo Results: We demonstrate surviving EGFR -mutant lung cancer cells upregulate the antiapoptotic protein MCL-1 in response to short-term EGFRi treatment. Mechanistically, DTCs undergo a protein biosynthesis enrichment resulting in increased mTORC1-mediated mRNA translation of MCL-1, revealing a novel mechanism in which lung cancer cells adapt to short-term pressures of apoptosis-inducing kinase inhibitors. Moreover, MCL-1 is a key molecule governing the emergence of early EGFR -mutant DTCs to EGFRi, and we demonstrate it can be effectively cotargeted with clinically emerging MCL-1 inhibitors both in vitro and in vivo Conclusions: Altogether, these data reveal that this novel therapeutic combination may delay the acquisition of secondary mutations, therefore prolonging therapy efficacy. Clin Cancer Res; 24(22); 5658-72. ©2018 AACR ., (©2018 American Association for Cancer Research.)

AMP-activated protein kinase (AMPK) is a master regulator of energy homeostasis and a promising drug target for managing metabolic diseases such as type 2 diabetes. Many pharmacological AMPK activators, and possibly unidentified physiological metabolites, bind to the allosteric drug and metabolite (ADaM) site at the interface between the kinase domain (KD) in the α-subunit and the carbohydrate-binding module (CBM) in the β-subunit. Here, using double electron-electron resonance (DEER) spectroscopy, we demonstrate that the CBM-KD interaction is partially dissociated and the interface highly disordered in the absence of pharmacological ADaM site activators as inferred from a low depth of modulation and broad DEER distance distributions. ADaM site ligands such as 991, and to a lesser degree phosphorylation, stabilize the KD-CBM association and strikingly reduce conformational heterogeneity in the ADaM site. Our findings that the ADaM site, formed by the KD-CBM interaction, can be modulated by diverse ligands and by phosphorylation suggest that it may function as a hub for integrating regulatory signals., (© 2018 Gu et al.)

High-risk neuroblastoma is often distinguished by amplification of MYCN and loss of differentiation potential. We performed high-throughput drug screening of epigenetic-targeted therapies across a large and diverse tumor cell line panel and uncovered the hypersensitivity of neuroblastoma cells to GSK-J4, a small-molecule dual inhibitor of lysine 27 of histone 3 (H3K27) demethylases ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX), and histone demethylase Jumonji D3 (JMJD3). Mechanistically, GSK-J4 induced neuroblastoma differentiation and endoplasmic reticulum (ER) stress, with accompanying up-regulation of p53 up-regulated modulator of apoptosis (PUMA) and induction of cell death. Retinoic acid (RA)-resistant neuroblastoma cells were sensitive to GSK-J4. In addition, GSK-J4 was effective at blocking the growth of chemorefractory and patient-derived xenograft models of high-risk neuroblastoma in vivo. Furthermore, GSK-J4 and RA combination increased differentiation and ER stress over GSK-J4 effects and limited the growth of neuroblastomas resistant to either drug alone. In MYCN -amplified neuroblastoma, PUMA induction by GSK-J4 sensitized tumors to the B cell lymphoma 2 (BCL-2) inhibitor venetoclax, demonstrating that epigenetic-targeted therapies and BCL-2 homology domain 3 mimetics can be rationally combined to treat this high-risk subset of neuroblastoma. Therefore, H3K27 demethylation inhibition is a promising therapeutic target to treat high-risk neuroblastoma, and H3K27 demethylation can be part of rational combination therapies to induce robust antineuroblastoma activity., (Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

We previously reported use of genotype surveillance data to predict outbreaks among incident tuberculosis clusters. We propose a method to detect possible outbreaks among endemic tuberculosis clusters. We detected 15 possible outbreaks, of which 10 had epidemiologic data or whole-genome sequencing results. Eight outbreaks were corroborated.

Purpose: Small-cell lung cancer (SCLC) is an often-fatal neuroendocrine carcinoma usually presenting as extensive disease, carrying a 3% 5-year survival. Despite notable advances in SCLC genomics, new therapies remain elusive, largely due to a lack of druggable targets. Experimental Design: We used a high-throughput drug screen to identify a venetoclax-sensitive SCLC subpopulation and validated the findings with multiple patient-derived xenografts of SCLC. Results: Our drug screen consisting of a very large collection of cell lines demonstrated that venetoclax, an FDA-approved BCL-2 inhibitor, was found to be active in a substantial fraction of SCLC cell lines. Venetoclax induced BIM-dependent apoptosis in vitro and blocked tumor growth and induced tumor regressions in mice bearing high BCL-2-expressing SCLC tumors in vivo BCL-2 expression was a predictive biomarker for sensitivity in SCLC cell lines and was highly expressed in a subset of SCLC cell lines and tumors, suggesting that a substantial fraction of patients with SCLC could benefit from venetoclax. Mechanistically, we uncover a novel role for gene methylation that helped discriminate high BCL-2-expressing SCLCs. Conclusions: Altogether, our findings identify venetoclax as a promising new therapy for high BCL-2-expressing SCLCs. Clin Cancer Res; 24(2); 360-9. ©2017 AACR ., (©2017 American Association for Cancer Research.)

We examined the National tuberculosis surveillance system to describe Hispanic persons who were incarcerated at time of tuberculosis (TB) diagnosis and to compare their characteristics with those of non-Hispanic incarcerated TB patients. After declines between 1993 and 2002, the annual proportion of Hispanic TB patients who were incarcerated grew from 4.9% in 2003 to 8.4% in 2014. During 2003-2014, 19% of incarcerated US-born TB patients were Hispanic, and 86% of the foreign-born were Hispanic. Most incarcerated TB patients were in local jails, but about a third of all foreign-born Hispanics were in the facility category that includes Immigration and Customs Enforcement detention centers. Foreign birth and recent U.S. arrival characterized many Hispanic persons receiving a TB diagnosis while incarcerated. Hispanic patients had twice the odds of being in federal prisons. Systematic efforts to identify TB infection and disease might lead to early diagnoses and prevention of future cases.

Treatment for latent tuberculous infection (LTBI) is a key strategy for the elimination of tuberculosis. Rare adverse reactions associated with LTBI treatment have been reported. We report the only case of acute kidney injury reported to Centers for Disease Control and Prevention surveillance for LTBI treatment-related adverse events. The patient experienced rapid intravascular hemolysis, resulting in heme pigment nephropathy; he was hospitalized and received three hemodialysis treatments, but recovered without sequelae. While LTBI treatment-related adverse events are rare, health care providers should maintain clinical vigilance and regularly counsel patients to facilitate prompt diagnoses and effective clinical management of affected patients.

Objectives: The Centers for Disease Control and Prevention provides on-site epidemiologic assistance for outbreak response when the health capacity of state, tribal, local, and territorial health departments has been exceeded. We examined recent outbreaks of tuberculosis (TB) for which health departments needed assistance., Methods: We defined a TB outbreak as detection of ≥3 TB cases related by transmission, as suggested by routine genotyping and epidemiologic linkages. We conducted retrospective reviews of documentation from all 21 TB outbreak investigations in the United States for which the Centers for Disease Control and Prevention provided on-site assistance during 2009-2015. We abstracted data on patients' demographic characteristics and TB risk factors, as well as factors contributing to the outbreak from trip reports written by on-site investigators, and we compared these with outbreaks investigated during 2002-2008., Results: The 21 TB outbreaks during 2009-2015 involved 457 outbreak patients (range, 3-99 patients per outbreak). Of the 21 outbreaks, 16 were first identified through genotype data. In sum, 118 (26%) patients were identified through contact investigations of other patients in the outbreak. Most outbreak patients (n = 363, 79%) were US born. Ninety-two (26%) patients had a mental illness, 204 (45%) had been homeless in the year before diagnosis, and 379 (83%) used alcohol excessively or used illicit substances. The proportion of patients experiencing homelessness doubled between 2002-2008 and 2009-2015; other characteristics were similar between the 2 periods. Delayed TB diagnosis contributed to unmitigated transmission in all but 1 outbreak., Conclusions: TB outbreaks challenge frontline public health resources. Genotyping and contact investigations are important strategies for detecting and controlling TB outbreaks, particularly among people experiencing homelessness or those with mental illness.

Objectives: Our objective was to describe and determine the factors contributing to a recent drug-resistant tuberculosis (TB) outbreak in Georgia., Methods: We defined an outbreak case as TB diagnosed from March 2008 through December 2015 in a person residing in Georgia at the time of diagnosis and for whom (1) the genotype of the Mycobacterium tuberculosis isolate was consistent with the outbreak strain or (2) TB was diagnosed clinically without a genotyped isolate available and connections were established to another outbreak-associated patient. To determine factors contributing to transmission, we interviewed patients and reviewed health records, homeless facility overnight rosters, and local jail booking records. We also assessed infection control measures in the 6 homeless facilities involved in the outbreak., Results: Of 110 outbreak cases in Georgia, 86 (78%) were culture confirmed and isoniazid resistant, 41 (37%) occurred in people with human immunodeficiency virus coinfection (8 of whom were receiving antiretroviral treatment at the time of TB diagnosis), and 10 (9%) resulted in TB-related deaths. All but 8 outbreak-associated patients had stayed overnight or volunteered extensively in a homeless facility; all these facilities lacked infection control measures. At least 9 and up to 36 TB cases outside Georgia could be linked to this outbreak., Conclusions: This article highlights the ongoing potential for long-lasting and far-reaching TB outbreaks, particularly among populations with untreated human immunodeficiency virus infection, mental illness, substance abuse, and homelessness. To prevent and control TB outbreaks, health departments should work with overnight homeless facilities to implement infection control measures and maintain searchable overnight rosters.

Serial femtosecond X-ray crystallography (SFX) using an X-ray free electron laser (XFEL) is a recent advancement in structural biology for solving crystal structures of challenging membrane proteins, including G-protein coupled receptors (GPCRs), which often only produce microcrystals. An XFEL delivers highly intense X-ray pulses of femtosecond duration short enough to enable the collection of single diffraction images before significant radiation damage to crystals sets in. Here we report the deposition of the XFEL data and provide further details on crystallization, XFEL data collection and analysis, structure determination, and the validation of the structural model. The rhodopsin-arrestin crystal structure solved with SFX represents the first near-atomic resolution structure of a GPCR-arrestin complex, provides structural insights into understanding of arrestin-mediated GPCR signaling, and demonstrates the great potential of this SFX-XFEL technology for accelerating crystal structure determination of challenging proteins and protein complexes.

G-protein-coupled receptors (GPCRs) signal primarily through G proteins or arrestins. Arrestin binding to GPCRs blocks G protein interaction and redirects signalling to numerous G-protein-independent pathways. Here we report the crystal structure of a constitutively active form of human rhodopsin bound to a pre-activated form of the mouse visual arrestin, determined by serial femtosecond X-ray laser crystallography. Together with extensive biochemical and mutagenesis data, the structure reveals an overall architecture of the rhodopsin-arrestin assembly in which rhodopsin uses distinct structural elements, including transmembrane helix 7 and helix 8, to recruit arrestin. Correspondingly, arrestin adopts the pre-activated conformation, with a ∼20° rotation between the amino and carboxy domains, which opens up a cleft in arrestin to accommodate a short helix formed by the second intracellular loop of rhodopsin. This structure provides a basis for understanding GPCR-mediated arrestin-biased signalling and demonstrates the power of X-ray lasers for advancing the frontiers of structural biology.

Small heterodimer partner (SHP) is an orphan nuclear receptor that functions as a transcriptional repressor to regulate bile acid and cholesterol homeostasis. Although the precise mechanism whereby SHP represses transcription is not known, E1A-like inhibitor of differentiation (EID1) was isolated as a SHP-interacting protein and implicated in SHP repression. Here we present the crystal structure of SHP in complex with EID1, which reveals an unexpected EID1-binding site on SHP. Unlike the classical cofactor-binding site near the C-terminal helix H12, the EID1-binding site is located at the N terminus of the receptor, where EID1 mimics helix H1 of the nuclear receptor ligand-binding domain. The residues composing the SHP-EID1 interface are highly conserved. Their mutation diminishes SHP-EID1 interactions and affects SHP repressor activity. Together, these results provide important structural insights into SHP cofactor recruitment and repressor function and reveal a conserved protein interface that is likely to have broad implications for transcriptional repression by orphan nuclear receptors.

MicroRNA-34a (miR-34a) is the most highly elevated hepatic miR in obese mice and is also substantially elevated in patients who have steatosis, but its role in obesity and metabolic dysfunction remains unclear. After a meal, FGF19 is secreted from the ileum; binds to a hepatic membrane receptor complex, FGF19 receptor 4 and coreceptor β-Klotho (βKL); and mediates postprandial responses under physiological conditions, but hepatic responses to FGF19 signaling were shown to be impaired in patients with steatosis. Here, we show an unexpected functional link between aberrantly elevated miR-34a and impaired βKL/FGF19 signaling in obesity. In vitro studies show that miR-34a down-regulates βKL by binding to the 3' UTR of βKL mRNA. Adenoviral-mediated overexpression of miR-34a in mice decreased hepatic βKL levels, impaired FGF19-activated ERK and glycogen synthase kinase signaling, and altered expression of FGF19 metabolic target genes. Consistent with these results, βKL levels were decreased and hepatic responses to FGF19 were severely impaired in dietary obese mice that have elevated miR-34a. Remarkably, in vivo antisense inhibition of miR-34a in obese mice partially restored βKL levels and improved FGF19 target gene expression and metabolic outcomes, including decreased liver fat. Further, anti-miR-34a treatment in primary hepatocytes of obese mice restored FGF19-activated ERK and glycogen synthase kinase signaling in a βKL-dependent manner. These results indicate that aberrantly elevated miR-34a in obesity attenuates hepatic FGF19 signaling by directly targeting βKL. The miR-34a/βKL/FGF19 axis may present unique therapeutic targets for FGF19-related human diseases, including metabolic disorders and cancer.

Abscisic acid (ABA) is an essential hormone that controls plant growth, development and responses to abiotic stresses. ABA signaling is mediated by type 2C protein phosphatases (PP2Cs), including HAB1 and ABI2, which inhibit stress-activated SnRK2 kinases and whose activity is regulated by ABA and ABA receptors. Based on biochemical data and our previously determined crystal structures of ABI2 and the SnRK2.6-HAB1 complex, we present the catalytic mechanism of PP2C and provide new insight into PP2C-SnRK2 interactions and possible roles of other SnRK2 kinases in ABA signaling.

Abscisic acid (ABA) is an essential hormone for plants to survive environmental stresses. At the center of the ABA signaling network is a subfamily of type 2C protein phosphatases (PP2Cs), which form exclusive interactions with ABA receptors and subfamily 2 Snfl-related kinase (SnRK2s). Here, we report a SnRK2-PP2C complex structure, which reveals marked similarity in PP2C recognition by SnRK2 and ABA receptors. In the complex, the kinase activation loop docks into the active site of PP2C, while the conserved ABA-sensing tryptophan of PP2C inserts into the kinase catalytic cleft, thus mimicking receptor-PP2C interactions. These structural results provide a simple mechanism that directly couples ABA binding to SnRK2 kinase activation and highlight a new paradigm of kinase-phosphatase regulation through mutual packing of their catalytic sites.

Abscisic acid (ABA) is a plant hormone that plays important roles in growth and development. ABA is also the central regulator to protect plants against abiotic stresses, such as drought, high salinity, and adverse temperatures, and ABA signaling is therefore a promising biotechnological target for the generation of crops with increased stress resistance. Recently, a core signal transduction pathway has been established, in which ABA receptors, type 2C protein phosphatases, and AMPK-related protein kinases control the regulation of transcription factors, ion channels, and enzymes. Here we use a simple protein thermal stability shift assay to independently validate key aspects of this pathway and to demonstrate the usefulness of this technique to detect and characterize very weak (Kd ≥ 50 µM) interactions between receptors and physiological and synthetic agonists, to determine and analyze protein-protein interactions, and to screen small molecule inhibitors.

Abscisic acid (ABA) is an essential hormone that controls plant growth, development, and responses to abiotic stresses. Central for ABA signaling is the ABA-mediated autoactivation of three monomeric Snf1-related kinases (SnRK2.2, -2.3, and -2.6). In the absence of ABA, SnRK2s are kept in an inactive state by forming physical complexes with type 2C protein phosphatases (PP2Cs). Upon relief of this inhibition, SnRK2 kinases can autoactivate through unknown mechanisms. Here, we report the crystal structures of full-length Arabidopsis thaliana SnRK2.3 and SnRK2.6 at 1.9- and 2.3-Å resolution, respectively. The structures, in combination with biochemical studies, reveal a two-step mechanism of intramolecular kinase activation that resembles the intermolecular activation of cyclin-dependent kinases. First, release of inhibition by PP2C allows the SnRK2s to become partially active because of an intramolecular stabilization of the catalytic domain by a conserved helix in the kinase regulatory domain. This stabilization enables SnRK2s to gain full activity by activation loop autophosphorylation. Autophosphorylation is more efficient in SnRK2.6, which has higher stability than SnRK2.3 and has well-structured activation loop phosphate acceptor sites that are positioned next to the catalytic site. Together, these data provide a structural framework that links ABA-mediated release of PP2C inhibition to activation of SnRK2 kinases.

Testicular receptors 2 and 4 (TR2/4) constitute a subgroup of orphan nuclear receptors that play important roles in spermatogenesis, lipid and lipoprotein regulation, and the development of the central nervous system. Currently, little is known about the structural features and the ligand regulation of these receptors. Here we report the crystal structure of the ligand-free TR4 ligand binding domain, which reveals an autorepressed conformation. The ligand binding pocket of TR4 is filled by the C-terminal half of helix 10, and the cofactor binding site is occupied by the AF-2 helix, thus preventing ligand-independent activation of the receptor. However, TR4 exhibits constitutive transcriptional activity on multiple promoters, which can be further potentiated by nuclear receptor coactivators. Mutations designed to disrupt cofactor binding, dimerization, or ligand binding substantially reduce the transcriptional activity of this receptor. Importantly, both retinol and retinoic acid are able to promote TR4 to recruit coactivators and to activate a TR4-regulated reporter. These findings demonstrate that TR4 is a ligand-regulated nuclear receptor and suggest that retinoids might have a much wider regulatory role via activation of orphan receptors such as TR4.

The phytohormone abscisic acid (ABA) functions through a family of fourteen PYR/PYL receptors, which were identified by resistance to pyrabactin, a synthetic inhibitor of seed germination. ABA activates these receptors to inhibit type 2C protein phosphatases, such as ABI1, yet it remains unclear whether these receptors can be antagonized. Here we demonstrate that pyrabactin is an agonist of PYR1 and PYL1 but is unexpectedly an antagonist of PYL2. Crystal structures of the PYL2-pyrabactin and PYL1-pyrabactin-ABI1 complexes reveal the mechanism responsible for receptor-selective activation and inhibition, which enables us to design mutations that convert PYL1 to a pyrabactin-inhibited receptor and PYL2 to a pyrabactin-activated receptor and to identify new pyrabactin-based ABA receptor agonists. Together, our results establish a new concept of ABA receptor antagonism, illustrate its underlying mechanisms and provide a rational framework for discovering novel ABA receptor ligands.

Transcription activation by androgen receptor (AR), which depends on recruitment of coactivators, is required for the initiation and progression of prostate cancer, yet the mechanisms of how hormone-activated AR interacts with coactivators remain unclear. This is because AR, unlike any other nuclear receptor, prefers its own N-terminal FXXLF motif to the canonical LXXLL motifs of coactivators. Through biochemical and crystallographic studies, we identify that steroid receptor coactivator-3 (SRC3) (also named as amplified in breast cancer-1 or AIB1) interacts strongly with AR via synergistic binding of its first and third LXXLL motifs. Mutagenesis and functional studies confirm that SRC3 is a preferred coactivator for hormone-activated AR. Importantly, AR mutations found in prostate cancer patients correlate with their binding potency to SRC3, corroborating with the emerging role of SRC3 as a prostate cancer oncogene. These results provide a molecular mechanism for the selective utilization of SRC3 by hormone-activated AR, and they link the functional relationship between AR and SRC3 to the development and growth of prostate cancer.

Abscisic acid (ABA) is a ubiquitous hormone that regulates plant growth, development and responses to environmental stresses. Its action is mediated by the PYR/PYL/RCAR family of START proteins, but it remains unclear how these receptors bind ABA and, in turn, how hormone binding leads to inhibition of the downstream type 2C protein phosphatase (PP2C) effectors. Here we report crystal structures of apo and ABA-bound receptors as well as a ternary PYL2-ABA-PP2C complex. The apo receptors contain an open ligand-binding pocket flanked by a gate that closes in response to ABA by way of conformational changes in two highly conserved beta-loops that serve as a gate and latch. Moreover, ABA-induced closure of the gate creates a surface that enables the receptor to dock into and competitively inhibit the PP2C active site. A conserved tryptophan in the PP2C inserts directly between the gate and latch, which functions to further lock the receptor in a closed conformation. Together, our results identify a conserved gate-latch-lock mechanism underlying ABA signalling.

A new multifunctional compliant instrument has been designed for use in minimally invasive surgery. The instrument combines scissors and forceps into a single multifunctional device. The main advantage of using multifunctional instruments for minimally invasive surgery is that instrument exchanges can be reduced, thus reducing procedure time and risk of inadvertent tissue injury during instrument exchanges. In this paper, the length, width, and thickness of the multifunctional compliant mechanism tool tip is optimized to maximize the jaw opening and the grasping force. The optimized design is then modeled to simulate the stresses encountered in the scissors mode. A 5.0 mm diameter stainless steel prototype is fabricated using electro-discharge machining and is shown to grasp and cut successfully.

The value of mailed educational videotapes as a means of enhancing compliance with drug therapy was studied. Members of a health maintenance organization with a pharmacy claim for benazepril, metoprolol, simvastatin, or transdermal estrogen were randomly assigned to a study group or a control group. Subjects in the study group were mailed one of four videotape programs giving information on the drug prescribed and the inferred disease state. Control subjects received no educational materials. Subjects were enrolled from July 1, 1993, through January 2, 1994. Refill data were collected from July 1, 1993, through April 1, 1994. The medication possession ratio (MPR) was calculated as the total number of days' supply of a drug obtained by a member divided by the number of days between the time of enrollment and April 1, 1994, or the date the member was terminated from the plan. A subject was deemed compliant if his or her MPR was > or = 0.80. There were no significant differences in mean MPRs between the study group (n = 1993) and the control group (n = 2253). None of the mean MPRs was > or = 0.80, although 44% of control subjects and 46% of study-group subjects were compliant. Of 97 respondents to a survey mailed to a randomly selected subset of the study group, almost 87% reported that they had viewed the videotapes, and of these subjects, about 88% said they found them very useful or somewhat useful. A one-time mailing of videotapes to patients, with no individual follow-up, did not increase compliance with the medications monitored.

Immunodeficient SCID (C.B-17 scid/scid) mice with persistent lactate dehydrogenase-elevating virus (LDV) infection failed to produce IgG anti-LDV antibodies, and during chronic infection transmitted virus infection to 95% of their offspring. In contrast, normal mice infected 15 or more days prior to giving birth produced IgG anti-LDV antibodies and transmitted LDV infection to only 0-46% of their fetuses. Transplacental transmission of LDV infection was dependent on the timing of maternal infection. Adoptive transfer of immune competence to LDV-infected SCID mice resulted in fetal protection from maternally transmitted virus infection. Fetal protection correlated with the presence of maternal IgG anti-LDV but not with fetal levels of IgG anti-LDV, and the levels of viremia in nonimmune SCID mice did not affect transplacental virus transmission. These results demonstrate the importance of maternal immunity in protecting the fetus from infection, and validate the use of this mouse model for investigation of immune mechanisms of transplacental virus transmission.

Whole lupins (Lupinus albus) were roasted with exit temperatures ranging from approximately 130 (moderate heat) to 175 degrees C (high heat). In situ N disappearance after 12 h of incubation in the rumen was 83% for raw lupins, 45% for lupins roasted at moderate temperatures, and 39% for lupins roasted at high temperatures. Lambs fed lupins roasted at moderate temperatures retained more N (P less than .01) than those fed soybean meal (SBM). However, growth rate and feed efficiency were similar among lambs fed diets containing SBM, raw lupins, or roasted lupins. Dehulled lupins commercially roasted at low, moderate, and high temperatures resulted in ruminal in situ N disappearances of 59, 47, and 43% for the respective temperatures. Dehulled lupins (Lupinus albus) were also roasted in a laboratory oven for 2, 4, and 6 h at 120, 140, and 160 degrees C. Simulation of roasting for 2 h had no effect (P greater than .10) on ruminal in situ N disappearance at any of the temperatures. In situ N disappearance was reduced (P less than .05) after roasting for 4 h at 160 degrees C, but acid detergent insoluble N was only moderately increased. Nitrogen retention in lambs fed raw, dehulled lupins was equal (P greater than .10) to that of lambs fed SBM. Whole lupins or dehulled lupins can replace SBM as the sole protein supplement for growing lambs. Although roasting lupins decreased ruminal in situ N disappearance, it had no effect on growth of lambs.

1. Glycoproteins of bovine (Bos taurus) and human (Homo sapiens) milk lipid globule membranes were characterized by ability to bind lectins after electrophoretic separation. 2. Seven lectin receptor glycoproteins were detected in bovine and five in human milk lipid globule membranes. Bovine and human globule membrane glycoproteins differed in ability to interact with certain lectins. 3. Two major nonionic detergent insoluble glycoproteins were present in bovine and human lipid globule membrane; these constituents had apparent molecular weights of 155,000 and 69,000. Detergent-insoluble polypeptides with similar or identical electrophoretic mobilities were found in milk lipid globule membranes from four other species, rat (Rattus norvegicus), sheep (Ovis aries), pig (Sus scrofa) and goat (Capra hircus). Tryptic peptide mapping revealed these polypeptides to be nonidentical among species.