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There is no previous study demonstrating the differences of genome-wide DNA methylation (DNAm) profiles between patients with and without postoperative delirium (POD). We aimed to discover epigenetic (DNAm) markers that are associated with POD in blood obtained from patients before and after neurosurgery.Pre- and post-surgical blood DNA samples from 37 patients, including 10 POD cases, were analyzed using the Illumina EPIC array genome-wide platform. We examined DNAm differences in blood from patients with and without POD. Enrichment analysis with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes terms were also conducted.When POD cases were tested for DNAm change before and after surgery, enrichment analyses showed many relevant signals with statistical significance in immune response related-pathways and inflammatory cytokine related-pathways such as "cellular response to cytokine stimulus", "regulation of immune system process", "regulation of cell activation", and "regulation of cytokine production". Furthermore, after excluding the potential effect of common factors related to surgery and anesthesia between POD cases and non-POD controls, the enrichment analyses showed significant signals such as "immune response" and "T cell activation", which are same pathways previously identified from an independent non-surgical inpatient cohort.Our first genome-wide DNAm investigation of POD showed promising signals related to immune response, inflammatory response and other relevant signals considered to be associated with delirium pathophysiology. Our data supports the hypothesis that epigenetics play an important role in the pathophysiological mechanism of delirium and suggest the potential usefulness of an epigenetics-based biomarker of POD.

AimsThere is no previous study demonstrating the differences of genome-wide DNA methylation (DNAm) profiles between patients with and without postoperative delirium (POD). We aimed to discover epigenetic (DNAm) markers that are associated with POD in blood obtained from patients before and after neurosurgery.MethodsPre- and post-surgical blood DNA samples from 37 patients, including 10 POD cases, were analyzed using the Illumina EPIC array genome-wide platform. We examined DNAm differences in blood from patients with and without POD. Enrichment analysis with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes terms were also conducted.ResultsWhen POD cases were tested for DNAm change before and after surgery, enrichment analyses showed many relevant signals with statistical significance in immune response related-pathways and inflammatory cytokine related-pathways such as “cellular response to cytokine stimulus”, “regulation of immune system process”, “regulation of cell activation”, and “regulation of cytokine production”. Furthermore, after excluding the potential effect of common factors related to surgery and anesthesia between POD cases and non-POD controls, the enrichment analyses showed significant signals such as “immune response” and “T cell activation”, which are same pathways previously identified from an independent non-surgical inpatient cohort. Conclusions: Our first genome-wide DNAm investigation of POD showed promising signals related to immune response, inflammatory response and other relevant signals considered to be associated with delirium pathophysiology. Our data supports the hypothesis that epigenetics are playing an important role in pathophysiological mechanism of delirium and suggest the potential usefulness of epigenetics based biomarker of POD.

Metformin has been reported to improve age-related disorders, including dementia, and to lower mortality. This study was conducted to investigate whether metformin use lower delirium risk, as well as long-term mortality.In this retrospective cohort study, previously recruited 1,404 subjects were analyzed. The relationship between metformin use and delirium, and the relationship between metformin use and 3-year mortality were investigated.242 subjects were categorized into a type 2 diabetes mellitus (DM)-without-metformin group, and 264 subjects were categorized into a DM-with-metformin group. Prevalence of delirium was 36.0% in the DM-without-metformin group, and 29.2% in the DM-with-metformin group. A history of metformin use reduced the risk of delirium in patients with DM (OR, 0.50 [95% CI, 0.32 to 0.79]) after controlling for confounding factors. The 3-year mortality in the DM-without-metformin group (survival rate, 0.595 [95% CI, 0.512 to 0.669]) was higher than in the DM-with-metformin group (survival rate, 0.695 [95% CI, 0.604 to 0.770]) (p=0.035). A history of metformin use decreased the risk of 3-year mortality after adjustment for confounding factors (HR, 0.69 [95% CI, 0.48 to 0.98]).Metformin use may lower the risk of delirium and mortality in DM patients.

ObjectiveTo investigate the relationship between history of metformin use and delirium risk, as well as long-term mortality.MethodsIn this retrospective cohort study, subjects recruited between January 2016 and March 2020 were analyzed. Logistic regression analysis was performed to investigate the relationship between metformin use and delirium. Log-rank analysis and Cox proportional hazards model were used to investigate the relationship between metformin use and 3-year mortality.ResultsThe data from 1404 subjects were analyzed. 242 subjects were categorized into a DM-without-metformin group, and 264 subjects were categorized into a DM-with-metformin group. Prevalence of delirium was 36.0% in the DM-without-metformin group, and 29.2% in the DM-with-metformin group. A history of metformin use reduced the risk of delirium in patients with DM (OR, 0.50 [95% CI, 0.32 to 0.79]) after controlling for age, sex, and dementia status, body mass index (BMI), and insulin use. The 3-year mortality in the DM-without-metformin group (survival rate, 0.595 [95% CI, 0.512 to 0.669]) was higher than in the DM-with-metformin group (survival rate, 0.695 [95% CI, 0.604 to 0.770]) (p=0.035). A history of metformin use decreased the risk of 3-year mortality after adjustment for age, sex, Charlson Comorbidity Index, BMI, history of insulin use, and delirium status (HR, 0.69 [95% CI, 0.48 to 0.98]).ConclusionsIt was found that metformin usage was associated with decreased delirium prevalence and lower 3-year mortality. The potential benefit of metformin on delirium risk and mortality were shown.

INTRODUCTION Lactate dehydrogenase A (LDHA) encodes an enzyme that catalyzes the inter-conversion between pyruvate and lactate in glycolysis. Here, we demonstrate that LDHA mediates a novel role in DNA repair independent of this metabolic function. METHODS siRNA screen, The Cancer Genome Atlas (TCGA) survival analysis, ionizing radiation (IR), g-H2AX, and chromatin assays, site-directed mutagenesis. RESULTS In an orthogonal siRNA-informatic screen to identify genes 1) when silenced caused IR sensitivity in patient-derived glioblastoma lines and 2) lowered expression is associated with improved survival in TCGA, LDHA surfaced as the top candidate. The survival association was validated by LDHA immunohistochemical staining in an independent collection of glioblastoma samples. In vitro and in vivo, silencing of LDHA sensitized glioblastoma lines to IR and enhanced radiation-induced g-H2AX accumulation. Such sensitization was not observed after treatment with an LDHA inhibitor, suggesting the metabolic function of LDHA is distinct from its role in DNA repair. Supporting this hypothesis, truncation mutations that suppressed the LDHA glycolysis function minimally affected its role in DNA repair. Mechanistically, cytoplasmic LDHA translocates into the nucleus in response to IR. This translocation was associated with subsequent chromatin transition into an open conformation and enhanced homologous recombination. CONCLUSION The novel LDHA function in DNA repair suggests intricate crosstalks between glycolytic metabolism and DNA repair, offering a new platform for glioblastoma therapeutic development.

We have previously developed a bispectral electroencephalography (BSEEG) device, which was shown to be effective in detecting delirium and predicting patient outcomes. In this study we aimed to apply the BSEEG approach for a sepsis. This was a retrospective cohort study conducted at a single center. Sepsis-positive cases were identified based on retrospective chart review. EEG raw data and calculated BSEEG scores were obtained in the previous studies. The relationship between BSEEG scores and sepsis was analyzed, as well as the relationship among sepsis, BSEEG score, and mortality. Data were analyzed from 628 patients. The BSEEG score from the first encounter (1st BSEEG) showed a significant difference between patients with and without sepsis (p = 0.0062), although AUC was very small indicating that it is not suitable for detection purpose. Sepsis patients with high BSEEG scores showed the highest mortality, and non-sepsis patients with low BSEEG scores showed the lowest mortality. Mortality of non-sepsis patients with high BSEEG scores was as bad as that of sepsis patients with low BSEEG scores. Even adjusting for age, gender, comorbidity, and sepsis status, BSEEG remained a significant predictor of mortality (p = 0.008). These data are demonstrating its usefulness as a potential tool for identification of patients at high risk and management of sepsis.

Background Recurrence after radiation therapy is nearly universal for glioblastomas, the most common form of adult brain cancer. The study aims to define clinically pertinent mechanisms underlying this recurrence. Methods microRNA (miRNA) profiling was performed using matched pre- and post-radiation treatment glioblastoma specimens from the same patients. All specimens harbored unmethylated O6-methylguanine-DNA methyltransferase promoters (umMGMT) and wild-type isocitrate dehydrogenase (wtIDH). The most altered miRNA, miR-603, was characterized. Findings While nearly all miRNAs remained unchanged after treatment, decreased levels of few, select miRNAs in the post-treatment specimens were observed, the most notable of which involved miR-603. Unbiased profiling of miR-603 targets revealed insulin-like growth factor 1 (IGF1) and IGF1 receptor (IGF1R). Ionizing radiation (IR) induced cellular export of miR-603 through extracellular vesicle (EV) release, thereby de-repressing IGF1 and IGF1R. This de-repression, in turn, promoted cancer stem-cell (CSC) state and acquired radiation resistance in glioblastomas. Export of miR-603 additionally de-repressed MGMT, a DNA repair protein responsible for detoxifying DNA alkylating agents, to promote cross-resistance to these agents. Ectopic miR-603 expression overwhelmed cellular capacity for miR-603 export and synergized with the tumoricidal effects of IR and DNA alkylating agents. Interpretation Profiling of matched pre- and post-treatment glioblastoma specimens revealed altered homeostasis of select miRNAs in response to radiation. Radiation-induced EV export of miR-603 simultaneously promoted the CSC state and up-regulated DNA repair to promote acquired resistance. These effects were abolished by exogenous miR-603 expression, suggesting potential for clinical translation. Funding NIH 1R01NS097649-01, 9R44GM128223-02, 1R01CA240953-01, the Doris Duke Charitable Foundation Clinical Scientist Development Award, The Sontag Foundation Distinguished Scientist Award, the Kimmel Scholar Award, and BWF 1006774.01 (C.C.C).

NASA is developing an expendable heavy lift launch vehicle capability, the Space Launch System, to support lunar and deep space exploration. To support this capability, an updated ground infrastructure is required including modifying an existing Mobile Launcher system. The Mobile Launcher is a very large heavy beam/truss steel structure designed to support the Space Launch System during its buildup and integration in the Vehicle Assembly Building, transportation from the Vehicle Assembly Building out to the launch pad, and provides the launch platform at the launch pad. The previous Saturn/Apollo and Space Shuttle programs had integrated vehicle ground vibration tests of their integrated launch vehicles performed with simulated free-free boundary conditions to experimentally anchor and validate structural and flight controls analysis models. For the Space Launch System program, the Mobile Launcher will be used as the modal test fixture for the ground vibration test of the first Space Launch System flight vehicle, Exploration Mission ? 1( now referred to as Artemis 1), programmatically referred to as the Integrated vehicle modal test. The Integrated vehicle modal test of the Exploration Mission - 1 integrated launch vehicle will have its core and second stages unfueled while mounted to the ML while inside the Vehicle Assembly Building, which is currently scheduled for the late spring or early summer of 2020. The Space Launch System program has implemented a building block approach for dynamic model validation. The modal test of the Mobile Launcher is an important part of this building block approach in supporting the integrated vehicle modal test since the Mobile Launcher will serve as a structurally dynamic test fixture whose modes will couple with the modes of the Exploration Mission ? 1 test vehicle. The Mobile Launcher modal test will further support understanding the structural dynamics of the Mobile Launcher and SLS during rollout to the launch pad, which will play a key role in better understanding and prediction of the rollout forces acting on the launch vehicle. The Mobile Launcher modal test is currently scheduled for the summer of 2019. Due to a very tight modal testing schedule, this Mobile Launcher modal pretest analysis has been performed to ensure there is a high likelihood of being able to successfully complete the modal test (i.e. identify the primary target modes) using the planned instrumentation, shakers, and excitation types. This paper will discuss this Mobile Launcher modal pretest analysis and the unique challenges faced due to the Mobile Launcher's size and weight, which are typically not faced when modal testing aerospace structures.

The Space Launch System (SLS) and its Mobile Launcher (ML) will be transported to the launch pad via the Crawler-Transporter (CT) system. Rollout (i.e., transportation) loads produce structural loads on the integrated SLS/Orion Multi-Purpose Crew Vehicle (MPCV) launch vehicle which are of a concern with respect to fatigue. As part of the risk reduction process and in addition to the modal building block test approach that has been adopted by the SLS Program, acceleration data will be obtained during rollout for use in modal parameter estimation. There are several occurrences where the ML/CT will be transported either into the Vertical Assembly Building (VAB) or to the launch pad and back without the SLS stack as part of the Kennedy Space Center (KSC) Exploration Ground Systems (EGS) Integrated Test and Checkout (ITCO). NASA KSC EGS has instrumentation installed on both the ML and CT to record data during rollout, at the launch pad, and during liftoff. The EGS instrumentation on the ML, which includes accelerometers, is referred to as the Sensor Data Acquisition System (SDAS). The EGS instrumentation on the CT, which also includes accelerometers, is referred to as the CT Data Acquisition System (CTDAS). The forces and accelerations applied to the ML and CT during a rollout event will be higher than any of the planned building block modal tests. This can be very beneficial in helping identify nonlinear behavior in the structure. Developing modal parameters from the same test hardware in multiple boundary conditions and under multiple levels of excitation is a key step in developing a well correlated FEM.

The p-nuclei are the few stable nuclei heavier than iron on the neutron-deficient side of the valley of stability that cannot be produced through astrophysical neutron-capture reactions. The limited experimental data on reactions through which the p-nuclei might be produced leaves the origin of their production largely unknown. This work presents the first cross section measurements of the 76Se( α , γ )80Kr reaction. The rate of the time reversed reaction, 80Kr( γ , α )76Se, is one of the most uncertain of possible reactions which can occur at the 80Kr branching point on the γ-process photo-disintegration pathway. The reaction flow through 80Kr will directly affect the final abundance of the p-nuclide 78Kr. Experimental cross sections at two astrophysically relevant energies are reported and compared to cross sections calculated using Hauser-Feshbach codes talys , non-smoker , and smaragd . The success of these first ( α , γ ) cross section measurements performed in inverse kinematics in the energy region of the γ-process opens the door for future studies of reactions on radioactive γ-process nuclides.

// Johnny C. Akers 1,* , Wei Hua 2,* , Hongying Li 3,* , Valya Ramakrishnan 1 , Zixiao Yang 2 , Kai Quan 2 , Wei Zhu 2 , Jie Li 1 , Javier Figueroa 1 , Brian R. Hirshman 1 , Brittney Miller 1 , David Piccioni 4 , Florian Ringel 5 , Ricardo Komotar 6 , Karen Messer 3 , Douglas R. Galasko 7 , Fred Hochberg 1 , Ying Mao 7,8,** , Bob S. Carter 1,** and Clark C. Chen 1,** 1 Center for Theoretical and Applied Neuro-Oncology, University of California, San Diego, CA, USA 2 Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China 3 Biostatistics Department, Moores Cancer Center, UC San Diego Health System, La Jolla, CA, USA 4 Department of Neurosurgery, Moores Cancer Center, UC San Diego Health System, La Jolla, CA, USA 5 Department of Neurosurgery, Klinikum rechts der Isar, Technische Universitat Munchen, Munich, Germany 6 Department of Neurological Surgery, Miller School of Medicine, University of Miami, Miami, FL, USA 7 Department of Neurosciences, University of California, San Diego, CA, USA 8 State Key Laboratory of Medical Neurobiology, Institutes of Brain Science, The Collaborative Innovation Center for Brain Science, Fudan University, Shanghai, China * These authors shared responsibility as first authors ** These authors shared responsibility as senior authors Correspondence to: Clark C. Chen, email: // Keywords : extracellular vesicle, CSF, liquid biopsy Received : April 12, 2017 Accepted : May 19, 2017 Published : June 01, 2017 Abstract Purpose: To develop a cerebrospinal fluid (CSF) miRNA diagnostic biomarker for glioblastoma. Experimental Design: Glioblastoma tissue and matched CSF from the same patient (obtained prior to tumor manipulation) were profiled by TaqMan OpenArray ® Human MicroRNA Panel. CSF miRNA profiles from glioblastoma patients and controls were created from three discovery cohorts and confirmed in two validation cohorts. Results: miRNA profiles from clinical CSF correlated with those found in glioblastoma tissues. Comparison of CSF miRNA profiles between glioblastoma patients and non-brain tumor patients yielded a tumor “signature” consisting of nine miRNAs. The “signature” correlated with glioblastoma tumor volume ( p =0.008). When prospectively applied to cisternal CSF, the sensitivity and specificity of the ‘signature’ for glioblastoma detection were 67% and 80%, respectively. For lumbar CSF, the sensitivity and specificity of the signature were 28% and 95%, respectively. Comparable results were obtained from analyses of CSF extracellular vesicles (EVs) and crude CSF. Conclusion: We report a CSF miRNA signature as a “liquid biopsy” diagnostic platform for glioblastoma.

During the slow neutron capture process in massive stars, reactions on light elements can both produce and absorb neutrons thereby influencing the final heavy element abundances. At low metallicities, the high neutron capture rate of 16-O can inhibit s-process nucleosynthesis unless the neutrons are recycled via the 17O(a,n)20Ne reaction. The efficiency of this neutron recycling is determined by competition between the 17O(a,n)20Ne and 17O(a,g)21Ne reactions. While some experimental data are available on the former reaction, no data exist for the radiative capture channel at the relevant astrophysical energies. The 17O(a,g)21Ne reaction has been studied directly using the DRAGON recoil separator at the TRIUMF Laboratory. The reaction cross section has been determined at energies between 0.6 and 1.6 MeV Ecm, reaching into the Gamow window for core helium burning for the first time. Resonance strengths for resonances at 0.63, 0.721, 0.81 and 1.122 MeV Ecm have been extracted. The experimentally based reaction rate calculated represents a lower limit, but suggests that significant s-process nucleosynthesis occurs in low metallicity massive stars., 7 pages, 5 figures

Studies on nuclear astrophysics, nuclear structure, and other interests have been performed using the radioactive-isotope (RI) beams at the low-energy RI beam separator CRIB, operated by Center for Nuclear Study (CNS), the University of Tokyo. The elastic resonant scattering is a striking tool to study astrophysical reactions and nuclear clusters. In particular, when it is coupled with a thick target and inverse kinematics, the measurement can be very efficient and even feasible with RI beams. By measuring resonant scattering, we can study the properties of resonant states which could play an important role in the astrophysical reaction, or have an exotic nuclear structure. The 10Be+α elastic scattering has been measured at CRIB, and three resonances which are in agreement with the prediction of a linear-chain structure by Suhara-En’yo were observed, giving a strong indication of the existence of such an exotic structure. Measurements based on the indirect technique of the reaction measurement, such as the Trojan horse method, have also been performed at CRIB.

Extracellular vesicles (EVs) are small, membrane-bound particles released by all cells that have emerged as an attractive biomarker platform. We study the utility of a dielectrophoretic (DEP) micro-chip device for isolation and characterization of EVs derived from plasma specimens from patients with brain tumors. EVs were isolated by DEP chip and subjected to on-chip immunofluorescence (IF) staining to determine the concentration of glial fibrillary acidic protein (GFAP) and Tau. EVs were analyzed from the plasma samples isolated from independent patient cohorts. Glioblastoma cell lines secrete EVs enriched for GFAP and Tau. These EVs can be efficiently isolated using the DEP platform. Application of DEP to clinical plasma samples afforded discrimination of plasma derived from brain tumor patients relative to those derived from patients without history of brain cancer. Sixty-five percent (11/17) of brain tumor patients showed higher EV-GFAP than the maximum observed in controls. Ninety-four percent (16/17) of tumor patients showed higher EV-Tau than the maximum observed in controls. These discrimination thresholds were applied to plasma isolated from a second, independent cohort of 15 glioblastoma patients and 8 controls. For EV-GFAP, we observed 93% sensitivity, 38% specificity, 74% PPV, 75% NPV, and AUC of 0.65; for EV-Tau, we found 67% sensitivity, 75% specificity 83% PPV, 55% NPV, and AUC of 0.71 for glioblastoma diagnosis. This proof-of-principle study provides support for DEP-IF of plasma EVs for diagnosis of glioblastoma.

Over the past 50 years, great advances have happened in both analytical modal analysis (i.e., finite element models and analysis) and experimental modal analysis (i.e., modal testing) in aerospace and other fields. With the advent of more powerful computers, higher performance instrumentation and data acquisition systems, and powerful linear modal extraction tools, today’s analysts and test engineers have a breadth and depth of technical resources only dreamed of by our predecessors. However, some observed recent trends indicate that hard lessons learned are being forgotten or ignored, and possibly fundamental concepts are not being understood. These trends have the potential of leading to the degradation of the quality of and confidence in both analytical and test results. These trends are a making of our own doing, and directly related to having ever more powerful computers, programmatic budgetary pressures to limit analysis and testing, and technical capital loss due to the retirement of the senior demographic component of a bimodal workforce. This paper endeavors to highlight some of the most important lessons learned, common pitfalls to hopefully avoid, and potential steps that may be taken to help reverse this trend.

Vibration testing spaceflight hardware is a vital, but time consuming and expensive endeavor. Traditionally modal tests are performed at the component, subassembly, or system level, preferably free-free with mass loaded interfaces or fixed base on a seismic mass to identify the fundamental structural dynamic (modal) characteristics. Vibration tests are then traditionally performed on single-axis slip tables at qualification levels that envelope the maximum predicted flight environment plus 3 dB and workmanship in order to verify the spaceflight hardware can survive its flight environment. These two tests currently require two significantly different test setups, facilities, and ultimately reconfiguration of the spaceflight hardware. The vision of this research is to show how traditional fixed-base modal testing can be accomplished using vibration qualification testing facilities, which not only streamlines testing and reduces test costs, but also opens up the possibility of performing modal testing to untraditionally high excitation levels that provide for test-correlated finite element models to be more representative of the spaceflight hardware’s response in a flight environment. This paper documents the first steps towards this vision, which is the comparison of modal parameters identified from a traditional fixed-based modal test performed on a modal floor and those obtained by utilizing a fixed based correction method with a large single-axis electrodynamic shaker driving a slip table supplemented with additional small portable shakers driving on the slip table and test article. To show robustness of this approach, the test article chosen is a simple linear weldment, whose mass, size, and modal parameters couple well with the dynamics of the shaker/slip table. This paper will show that all dynamics due to the shaker/slip table were successfully removed resulting in true fixed-base modal parameters, including modal damping, being successfully extracted from a traditional style base-shake vibration test setup.

The Orion European Service Module—Structural Test Article (E-STA) underwent sine vibration testing in 2016 using the Mechanical Vibration Facility (MVF) multi-axis shaker system at NASA Glenn Research Center’s (GRC) Plum Brook Station (PBS) Space Power Facility (SPF). The main objective was to verify the structural integrity of the European Service Module (ESM) under sine sweep dynamic qualification vibration testing. A secondary objective was to perform a fixed-base modal survey, while E-STA was still mounted to MVF, in order to achieve a test correlate the finite element model (FEM). To facilitate the E-STA system level correlation effort, a building block test approach was implemented. Modal tests were performed on two major subassemblies, the crew module/launch abort structure (CM/LAS) and the crew module adapter (CMA) mass simulators. These subassembly FEMs were individually correlated and then integrated into the E-STA FEM prior to the start of the E-STA sine vibration test. This paper summarizes the modal testing and model correlation efforts of both of these subassemblies and how the building block approach assisted in the overall correlation of the E-STA FEM. This paper will also cover modeling practices that should be avoided, recommended instrumentation positioning on complex structures, and the importance of the FEM geometrically matching CAD in sufficient detail in order to adequately replicate internal load paths. The goal of this paper is to inform the reader of the hard earned lessons learned and pitfalls to avoid when applying a building block test approach.

Background Tumor specific genetic material can be detected in extracellular vesicles (EVs) isolated from blood, cerebrospinal fluid (CSF), and other biofluids of glioblastoma patients. As such, EVs have emerged as a promising platform for biomarker discovery. However, the optimal procedure to transport clinical EV samples remains poorly characterized. Methods We examined the stability of EVs isolated from CSF of glioblastoma patients that were stored under different conditions. EV recovery was determined by Nanoparticle tracking analysis, and qRT-PCR was performed to determine the levels of miRNAs. Results CSF EVs that were lyophilized and stored at room temperature (RT) for seven days exhibited a 37-43% reduction in EV number. This reduction was further associated with decreased abundance of representative miRNAs. In contrast, the EV number and morphology remained largely unchanged if CSF were stored at RT. Total RNA and representative miRNA levels were well-preserved under this condition for up to seven days. A single cycle of freezing and thawing did not significantly alter EV number, morphology, RNA content, or miRNA levels. However, incremental decreases in these parameters were observed after two cycles of freezing and thawing. Conclusions These results suggest that EVs in CSF are stable at RT for at least seven days. Repeated cycles of freezing/thawing should be avoided to minimize experimental artifacts.

KOBRA (KOrea Broad acceptance Recoil spectrometer and Apparatus) facility being designed at Rare Isotope Science Project in Korea will be utilized to produce rare isotope beams by employing multi-nucleon transfer reactions at about 20 MeV/nucleon for studies of nuclear structure. KOBRA will also provide high suppression of beam induced background for direct measurements of radiative-capture cross sections in the astrophysical energy range. The present design status of the KOBRA facility is reported along with a brief introduction to the facility. We have studied the feasibility of production of 44 Ti based on the present design of KOBRA as an example, and calculated the intensity of 44 Ti secondary beam, to be about 105 particles per second, for 1 pnA 46 Ti primary beam with a carbon target for a beam energy of 25 MeV/nucleon. A Monte Carlo simulation with a ray-tracing code has been performed to show that recoil products 66 Se are well separated from a 65 As beam by KOBRA for the 65 As ( p , γ ) 66 Se reaction at a beam energy of 1 MeV/nucleon.

Transfusion safety coordinators (TSCs) are an integral part of the transfusion process, which involves many interlinking chains of events and a multidisciplinary group of health professionals. In Australia, individual hospital-based TSCs have been in place for several decades, with state-based collaboratives commencing in 2002. The role has expanded across the country and currently there are 113 dedicated TSC positions and many more staff involved as blood/transfusion champions. There are also 12 transfusion nurse (TN) positions within the Australian Red Cross Blood Service. Over time both TSC and TN roles have evolved to meet the changes within the Australian blood sector. The primary focus of safety and appropriateness has now evolved to be more patient-centred by incorporating patient blood management (PBM) initiatives. National PBM guidelines, statements, strategies, criteria and healthcare standards specifically focused on all aspects of transfusion have influenced this evolution. TSCs undertake diverse roles and activities that vary significantly between health services and within each jurisdiction. Effective communication and change management skills are integral to the success of the role. Conclusion: The TSCs are highly recognised within the transfusion team and the role continues to evolve with the changes in the Australian blood sector. The term Transfusion Safety Coordinators (TSCs) has been used to describe roles such as transfusion nurse/trainer/practitioner/safety officer/clinical nurse consultant and PBM nurses.

INTRODUCTION Release of exosomes and extracellular vesicles (EV) by glioblastomas plays key roles in modulating the tumor microenvironment and therapeutic response. Studies to date have largely focused on the impact of EV and exosomes on the recipient cell. Here, we demonstrate that EV-mediated export of a master-regulatory miRNA has fate-determining impacts on the cell releasing the EVs. METHODS microRNA (miRNA) profiling was performed using clinical glioblastoma specimens from the same patients derived pre- and post-standard of care treatment. Mechanism mediating altered miRNA homeostasis were assessed. RESULTS While the levels of nearly all miRNAs remained unchanged after standard-of-care treatment, decreased levels of few, select miRNAs were observed, including miR-603. In response to ionizing radiation (IR), but not temozolomide (TMZ), glioblastoma cell lines exhibited a time-dependent decrease in miR-603 levels. While miR-603 biogenesis and degradation remained unchanged after IR, IR induced an increase in EV-mediated export of miR-603. Profiling of miR-603 targets revealed that miR-603 repressed the insulin-like growth factor 1 (IGF1) and IGF1 receptor (IGF1R), genes required to maintain the cancer stem cell (CSC) state, as well as MGMT, the gene encoding a DNA repair protein that detoxifies temozolomide (TMZ) induced DNA damages. IR induced export of miR-603 de-repress IGF1/IGF1R to promote radiation resistance and de-repress MGMT to promote cross-resistance to TMZ and CCNU. Ectopic miR-603 expression overwhelmed cellular capacity for miR-603 export and synergized with the tumoricidal effects of IR and temozolomide (TMZ). CONCLUSIONS Radiation stimulated EV-mediated export of miR-603 to facilitate acquired resistance to IR and cross-resistance to DNA alkylating agents.

Temozolomide (TMZ), the standard-of-care chemotherapy for glioblastoma, is a DNA alkylating drug. At cytotoxic doses, TMZ Induces high levels of alkylated DNA, including O6 methyl-guanine (O6MG), ultimately triggering cell death. In sub-lethal levels, O6MGs undergo non-Watson-Crick pairing, facilitating mutagenesis and promote acquired resistance. Whether the cytotoxic or mutagenic effects of TMZ predominate is largely determined by the DNA repair enzyme, Methyl-guanine-methyl transferase (MGMT). MGMT restores TMZ-alkylated DNA into its native form. To date, studies of MGMT have largely focused on its expression in large populations of cells. Variation of MGMT expression at a single cell level and pertinence of this variation to acquired TMZ resistance remains poorly understood. In single cells derived from the same patient-derived glioblastoma line, we found that the cell-to-cell variability in MGMT mRNA expression differed by as much as 10-fold. The distribution of single cell MGMT expression approximated that of a normal distribution. Upon treatment with TMZ, the mean expression of MGMT increased by two-fold. Additionally, TMZ treatment increased the standard deviation of the MGMT expression distribution by ten-fold, causing significant “widening” of the range the MGMT expression. Both of these effects were caused by degradation of miR-181d, an MGMT suppressing miRNA. TMZ treatment triggered the activation of ATM- and Rad3- related (ATR) kinase, which in turn activated Polyribonucleotide nucleotidyltransferase 1 (PNPT1), an RNAse that ultimately degradation of miR-181d. Our results suggest that RNA degradation enhanced stochastic variation in MGMT expression, allowing a wider range of MGMT expression from which the optimally fit clone can emerge. This novel form of TMZ resistance can be suppressed by over-expression of miR-181d, suggesting the potential of microRNA delivery as a therapeutic strategy. Accordingly, miR-181d delivery combined with TMZ treatment effectively cured intracranial murine models of patient derived glioblastoma xenografts (PDX) that expressed high levels of MGMT.

In 2016, the Orion European Service Module Structural Test Article (E-STA) underwent sine vibration testing using the multi-axis shaker system at NASA Glenn Research Center’s (GRC) Plum Brook Station (PBS) Space Power Facility (SPF) Mechanical Vibration Facility (MVF). An innovative approach using measured constraint shapes at the interface of E-STA to the MVF Table allowed high-quality fixed base modal parameters of the E-STA to be extracted, which have been used to update the E-STA finite element model (FEM), without the need for a traditional fixed base modal survey. This innovative approach provided considerable program cost and test schedule savings. This paper documents this modal survey, which includes the modal pretest analysis sensor selection, the fixed base methodology using measured constraint shapes as virtual references and measured frequency response functions, and post-survey comparison between measured and analysis fixed base modal parameters.

Exosomes can mediate a dynamic method of communication between malignancies, including those sequestered in the central nervous system and the immune system. We sought to determine whether exosomes from glioblastoma (GBM)-derived stem cells (GSCs) can induce immunosuppression. We report that GSC-derived exosomes (GDEs) have a predilection for monocytes, the precursor to macrophages. The GDEs traverse the monocyte cytoplasm, cause a reorganization of the actin cytoskeleton, and skew monocytes toward the immune suppresive M2 phenotype, including programmed death-ligand 1 (PD-L1) expression. Mass spectrometry analysis demonstrated that the GDEs contain a variety of components, including members of the signal transducer and activator of transcription 3 (STAT3) pathway that functionally mediate this immune suppressive switch. Western blot analysis revealed that upregulation of PD-L1 in GSC exosome-treated monocytes and GBM-patient-infiltrating CD14+ cells predominantly correlates with increased phosphorylation of STAT3, and in some cases, with phosphorylated p70S6 kinase and Erk1/2. Cumulatively, these data indicate that GDEs are secreted GBM-released factors that are potent modulators of the GBM-associated immunosuppressive microenvironment.

Background: Nucleosynthesis of mid-mass elements is thought to occur under hot and explosive astrophysical conditions. Radiative α capture on S34 has been shown to impact nucleosynthesis in several such conditions, including core and shell oxygen burning, explosive oxygen burning, and type Ia supernovae. Purpose: Broad uncertainties exist in the literature for the strengths of three resonances within the astrophysically relevant energy range (ECM=1.94-3.42MeV at T=2.2GK). Further, there are several states in Ar38 within this energy range which have not been previously measured. This work aimed to remeasure the resonance strengths of states for which broad uncertainty existed as well as to measure the resonance strengths and energies of previously unmeasured states. Methods: Resonance strengths and energies of eight narrow resonances (five of which had not been previously studied) were measured in inverse kinematics with the DRAGON facility at TRIUMF by impinging an isotopically pure beam of S34 ions on a windowless He4 gas target. Prompt γ emissions of de-exciting Ar38 recoils were detected in an array of bismuth germanate scintillators in coincidence with recoil nuclei, which were separated from unreacted beam ions by an electromagnetic mass separator and detected by a time-of-flight system and a multianode ionization chamber. Results: The present measurements agree with previous results. Broad uncertainty in the resonance strength of the ECM=2709keV resonance persists. Resonance strengths and energies were determined for five low-energy resonances which had not been studied previously, and their strengths were determined to be significantly weaker than those of previously measured resonances. Conclusions: The five previously unmeasured resonances were found not to contribute significantly to the total thermonuclear reaction rate. A median total thermonuclear reaction rate calculated using data from the present work along with existing literature values using the STARLIB rate calculator agrees with the NON-SMOKER statistical model calculation as well as the REACLIB and STARLIB library rates at explosive and nonexplosive oxygen-burning temperatures (T=3-4GK and T=1.5-2.7GK, respectively).

Background: Classical novae are cataclysmic nuclear explosions occurring when a white dwarf in a binary system accretes hydrogen-rich material from its companion star. Novae are partially responsible for the galactic synthesis of a variety of nuclides up to the calcium ( A ∼ 40 ) region of the nuclear chart. Although the structure and dynamics of novae are thought to be relatively well understood, the predicted abundances of elements near the nucleosynthesis endpoint, in particular Ar and Ca, appear to sometimes be in disagreement with astronomical observations of the spectra of nova ejecta. Purpose: One possible source of the discrepancies between model predictions and astronomical observations is nuclear reaction data. Most reaction rates near the nova endpoint are estimated only from statistical model calculations, which carry large uncertainties. For certain key reactions, these rate uncertainties translate into large uncertainties in nucleosynthesis predictions. In particular, the 38 K ( p , γ ) 39 Ca reaction has been identified as having a significant influence on Ar, K, and Ca production. In order to constrain the rate of this reaction, we have performed a direct measurement of the strengths of three candidate l = 0 resonances within the Gamow window for nova burning, at 386 ± 10 keV, 515 ± 10 keV, and 689 ± 10 keV. Method: The experiment was performed in inverse kinematics using a beam of unstable 38 K impinged on a windowless hydrogen gas target. The 39 Ca recoils and prompt γ rays from 38 K ( p , γ ) 39 Ca reactions were detected in coincidence using a recoil mass separator and a bismuth-germanate scintillator array, respectively. Results: For the 689 keV resonance, we observed a clear recoil- γ coincidence signal and extracted resonance strength and energy values of 120 + 50 − 30 ( stat . ) + 20 − 60 ( sys . ) meV and 679 + 2 − 1 ( stat . ) ± 1 ( sys . ) keV , respectively. We also performed a singles analysis of the recoil data alone, extracting a resonance strength of 120 ± 20 ( stat . ) ± 15 ( sys . ) meV, consistent with the coincidence result. For the 386 keV and 515 keV resonances, we extract 90 % confidence level upper limits of 2.54 meV and 18.4 meV, respectively. Conclusions: We have established a new recommended 38 K ( p , γ ) 39 Ca rate based on experimental information, which reduces overall uncertainties near the peak temperatures of nova burning by a factor of ∼ 250 . Using the rate obtained in this work in model calculations of the hottest oxygen-neon novae reduces overall uncertainties on Ar, K, and Ca synthesis to factors of 15 or less in all cases.

International audience; Studies on nuclear astrophysics, nuclear structure, and other interests have been performed using the radioactive-isotope (RI) beams at the low-energy RI beam separator CRIB, operated by Center for Nuclear Study (CNS), the University of Tokyo. A type of measurement to study astophysical reactions at CRIB is by the elastic resonant scattering with the thick-target method in inverse kinematics. An example is the α resonant scattering with 7Be beam, related to the astrophysical 7Be(α,γ) reactions, which is relevant in the hot p-p chain and νp-process in supernovae. Other α resonant scattering measurements with 30S, 10Be, 15O, and 18Ne beams have been performed at CRIB, using the thick-target method. There have also been measurements based on other experimental methods. The first Trojan horse method (THM) measurement using an RI beam has been performed at CRIB, to study the 18F(p, α)15O reaction at astrophysical energies via the three body reaction 2H(18F, α15O)n. The 18F(p, α)15O reaction rate is crucial to understand the 511-keV γ-ray production in nova explosion phenomena, and we successfully evaluated the reaction cross section at novae temperature and below experimentally for the first time.

We have performed a direct measurement of the 19Ne(p,γ)20Na reaction in inverse kinematics using a beam of radioactive 19Ne. The key astrophysical resonance in the 19Ne+p system has been definitely measured for the first time at Ec.m.=456+5−2 keV with an associated strength of 17+7−5 meV. The present results are in agreement with resonance strength upper limits set by previous direct measurements, as well as resonance energies inferred from precision (3He, t) charge exchange reactions. However, both the energy and strength of the 456 keV resonance disagree with a recent indirect study of the 19Ne(d, n)20Na reaction. In particular, the new 19Ne(p,γ)20Na reaction rate is found to be factors of ∼8 and ∼5 lower than the most recent evaluation over the temperature range of oxygen-neon novae and astrophysical x-ray bursts, respectively. Nevertheless, we find that the 19Ne(p,γ)20Na reaction is likely to proceed fast enough to significantly reduce the flux of 19F in nova ejecta and does not create a bottleneck in the breakout from the hot CNO cycles into the rp process.

We have performed a direct measurement of the ^{19}Ne(p,γ)^{20}Na reaction in inverse kinematics using a beam of radioactive ^{19}Ne. The key astrophysical resonance in the ^{19}Ne+p system has been definitely measured for the first time at E_{c.m.}=456_{-2}^{+5} keV with an associated strength of 17_{-5}^{+7} meV. The present results are in agreement with resonance strength upper limits set by previous direct measurements, as well as resonance energies inferred from precision (^{3}He, t) charge exchange reactions. However, both the energy and strength of the 456 keV resonance disagree with a recent indirect study of the ^{19}Ne(d, n)^{20}Na reaction. In particular, the new ^{19}Ne(p,γ)^{20}Na reaction rate is found to be factors of ∼8 and ∼5 lower than the most recent evaluation over the temperature range of oxygen-neon novae and astrophysical x-ray bursts, respectively. Nevertheless, we find that the ^{19}Ne(p,γ)^{20}Na reaction is likely to proceed fast enough to significantly reduce the flux of ^{19}F in nova ejecta and does not create a bottleneck in the breakout from the hot CNO cycles into the rp process.

Tauopathies are a class of neurodegenerative diseases, including Alzheimer's disease, frontotemporal dementia and progressive supranuclear palsy, which are associated with the pathological aggregation of tau protein into neurofibrillary tangles (NFT). Studies have characterized tau as a "prion-like" protein given its ability to form distinct, stable amyloid conformations capable of transcellular and multigenerational propagation in clonal fashion. It has been proposed that progression of tauopathy could be due to the prion-like propagation of tau, suggesting the possibility that end-stage pathologies, like NFT formation, may require an instigating event such as tau seeding. To investigate this, we applied a novel human induced pluripotent stem cell (hiPSC) system we have developed to serve as a human neuronal model. We introduced the tau repeat domain (tau-RD) with P301L and V337M (tau-RD-LM) mutations into hiPSC-derived neurons and observed expression of tau-RD at levels similar to total tau in postmortem AD brains. Tau aggregation occurred without the addition of recombinant tau fibrils. The conditioned media from tau-RD cultures contained tau-RD seeds, which were capable of inducing aggregate formation in homotypic mode in non-transduced recipient neuronal cultures. The resultant NFTs were thioflavin-positive, silver stain-positive, and assumed fibrillary appearance on transmission electron microscopy (TEM) with immunogold, which revealed paired helical filament 1 (PHF1)-positive NFTs, representing possible recruitment of endogenous tau in the aggregates. Functionally, expression of tau-RD caused neurotoxicity that manifested as axon retraction, synaptic density reduction, and enlargement of lysosomes. The results of our hiPSC study were reinforced by the observation that Tau-RD-LM is excreted in exosomes, which mediated the transfer of human tau to wild-type mouse neurons in vivo. Our hiPSC human neuronal system provides a model for further studies of tau aggregation and pathology as well as a means to study transcellular propagation and related neurodegenerative mechanisms.

We examined the extracellular vesicle (EV) and RNA composition of pooled normal cerebrospinal fluid (CSF) samples and CSF from five major neurological disorders: Alzheimer’s disease (AD), Parkinson’s disease (PD), low-grade glioma (LGG), glioblastoma multiforme (GBM), and subarachnoid haemorrhage (SAH), representing neurodegenerative disease, cancer, and severe acute brain injury. We evaluated: (I) size and quantity of EVs by nanoparticle tracking analysis (NTA) and vesicle flow cytometry (VFC), (II) RNA yield and purity using four RNA isolation kits, (III) replication of RNA yields within and between laboratories, and (IV) composition of total and EV RNAs by reverse transcription–quantitative polymerase chain reaction (RT-qPCR) and RNA sequencing (RNASeq). The CSF contained ~106 EVs/μL by NTA and VFC. Brain tumour and SAH CSF contained more EVs and RNA relative to normal, AD, and PD. RT-qPCR and RNASeq identified disease-related populations of microRNAs and messenger RNAs (mRNAs) relative to normal CSF, in both total and EV fractions. This work presents relevant measures selected to inform the design of subsequent replicative CSF studies. The range of neurological diseases highlights variations in total and EV RNA content due to disease or collection site, revealing critical considerations guiding the selection of appropriate approaches and controls for CSF studies.

Background RNAs within extracellular vesicles (EVs) have potential as diagnostic biomarkers for patients with cancer and are identified in a variety of biofluids. Glioblastomas (GBMs) release EVs containing RNA into cerebrospinal fluid (CSF). Here we describe a multi-institutional study of RNA extracted from CSF-derived EVs of GBM patients to detect the presence of tumor-associated amplifications and mutations in epidermal growth factor receptor (EGFR). Methods CSF and matching tumor tissue were obtained from patients undergoing resection of GBMs. We determined wild-type (wt)EGFR DNA copy number amplification, as well as wtEGFR and EGFR variant (v)III RNA expression in tumor samples. We also characterized wtEGFR and EGFRvIII RNA expression in CSF-derived EVs. Results EGFRvIII-positive tumors had significantly greater wtEGFR DNA amplification (P = 0.02) and RNA expression (P = 0.03), and EGFRvIII-positive CSF-derived EVs had significantly more wtEGFR RNA expression (P = 0.004). EGFRvIII was detected in CSF-derived EVs for 14 of the 23 EGFRvIII tissue-positive GBM patients. Conversely, only one of the 48 EGFRvIII tissue-negative patients had the EGFRvIII mutation detected in their CSF-derived EVs. These results yield a sensitivity of 61% and a specificity of 98% for the utility of CSF-derived EVs to detect an EGFRvIII-positive GBM. Conclusion Our results demonstrate CSF-derived EVs contain RNA signatures reflective of the underlying molecular genetic status of GBMs in terms of wtEGFR expression and EGFRvIII status. The high specificity of the CSF-derived EV diagnostic test gives us an accurate determination of positive EGFRvIII tumor status and is essentially a less invasive "liquid biopsy" that might direct mutation-specific therapies for GBMs.

While mutations in isocitrate dehydrogenase (IDH) gene are associated with favorable prognosis in glioblastoma patients, the biologic basis for favorable survival in patients with wild-type IDH (wtIDH) glioblastoma remains poorly understood. We identified that an inflammatory gene signature whose expression inversely correlated with patient survival in three independent, wtIDH glioblastoma cohorts (The Cancer Genome Atlas (TCGA), Chinese Glioma Genome Atlas (CGGA), and REMEBRANDT). Systematic analysis of the inflammatory infiltrates of the glioblastoma microenvironment revealed microglia as the predominant cell type driving this survival association. The pro-tumorgenic effects of microglia can be recapitulated using conditioned media derived from microglia. We cross-referenced proteomic profile of microglia conditioned media with the TCGA and the CGGA to identify constituent soluble factors with survival association and identified IL11. In both clinical datasets, increased IL11 expression was associated with shortened survival. Moreover, IL11 expression closely correlated with the survival-associated inflammatory signature in both TCGA and CGGA glioblastomas. IL11 mRNA and protein expression are elevated in clinical glioblastoma specimen relative to the surrounding cerebrum. Analysis of samples secured from three unrelated patients revealed that IL11 expression was largely found in the microglia fraction. In vitro and in vivo experiments demonstrate that IL11 was necessary and sufficient for enhancement of glioblastoma tumorigenicity. Microglia released IL11 induced activation of a STAT3-MYC signaling axis in glioblastoma cells, which ultimately lead to up-regulation of genes required for glioblastoma tumorigenicity, including OLIG2, SOX2, and POU3F2. PI3Kγ activation in myeloid-derived cells, including microglia, is essential for its trafficking into the tumor microenvironment. We hypothesized that PI3Kγ inhibition or inactivation should impede microglia trafficking to the glioblastoma microenvironment and suppress tumorigenicity. Supporting our hypothesis, treatment with the PI3Kγ, TG100-115, suppressed glioblastoma tumorigenicity in vivo by reduced microglia density and IL11 release in the glioblastoma microenvironment. These effects were recapitulated when murine glioblastoma GL261 were implanted into p110γ-/- mice. Importantly, ectopic expression IL11 in GL261 reversed the tumor-suppressive effect of TG100-115. The anti-glioblastoma effects of TG100-115 were enhanced by temozolomide, the standard of care chemotherapy for glioblastoma. These results suggest microglia is a mediator of clinical glioblastoma survival and disruption of microglia-glioblastoma interaction as potential therapeutic strategy.