Evaluation of plasma nucleosome concentrations in dogs with a variety of common cancers and in healthy dogs.

BACKGROUND: Cell free DNA, in the form of nucleosomes, is released into circulation during apoptosis and necrosis in a variety of diseases. They are small fragments of chromosomes that are composed of DNA wrapped around a histone core made of four duplicate histone proteins forming an octamer. The nucleosome compartment is a relatively uninvestigated area of circulating tumor biomarkers in dogs. The objectives of this study were to quantify and better characterize nucleosome concentrations in 528 dogs with various common malignancies and compare them to 134 healthy dogs. RESULTS: The sensitivity of increased circulating nucleosome concentrations for the detection of cancer in all dogs was 49.8% with a specificity of 97% with an area under the curve of 68.74%. The top 4 malignancies detected by the test included lymphoma, hemangiosarcoma, histiocytic sarcoma and malignant melanoma. The malignancies least likely to be detected were soft tissue sarcomas, osteosarcoma and mast cell tumors. CONCLUSIONS: A variety of tumor types may cause increased nucleosome concentrations in dogs. Tumors of hematopoietic origin are most likely to cause elevations and local tumors such as soft tissue sarcomas are least likely to cause elevations in plasma nucleosome concentrations.

Sub-Kelvin cooling for two kilopixel bolometer arrays in the PIPER receiver.

The Primordial Inflation Polarization Explorer (PIPER) is a balloon-borne telescope mission to search for inflationary gravitational waves from the early universe. PIPER employs two 32 × 40 arrays of superconducting transition-edge sensors, which operate at 100 mK. An open bucket Dewar of liquid helium maintains the receiver and telescope optics at 1.7 K. We describe the thermal design of the receiver and sub-Kelvin cooling with a continuous adiabatic demagnetization refrigerator (CADR). The CADR operates between 70 and 130 mK and provides ≈10 µW cooling power at 100 mK, nearly five times the loading of the two detector assemblies. We describe electronics and software to robustly control the CADR, overall CADR performance in flightlike integrated receiver testing, and practical considerations for implementation in the balloon float environment.

Tau positron emission tomography imaging in C9orf72 repeat expansion carriers.

BACKGROUND AND PURPOSE: AV-1451 (18 F-AV-1451, flortaucipir) positron emission tomography was performed in C9orf72 expansion carriers to assess tau accumulation and disease manifestation. METHODS: Nine clinically characterized C9orf72 expansion carriers and 18 age- and gender- matched cognitively normal individuals were psychometrically evaluated and underwent tau positron emission tomography imaging. The regional AV-1451 standard uptake value ratios from multiple brain regions were analyzed. Spearman correlation was performed to relate the AV-1451 standard uptake value ratio to clinical, psychometric and cerebrospinal fluid measures. RESULTS: C9orf72 expansion carriers had increased AV-1451 binding in the entorhinal cortex compared to controls. Primary age-related tauopathy was observed postmortem in one patient. AV-1451 uptake did not correlate with clinical severity, disease duration, psychometric performance or cerebrospinal fluid markers. CONCLUSION: C9orf72 expansion carriers exhibited increased AV-1451 uptake in entorhinal cortex compared to cognitively normal controls, suggesting a propensity for primary age-related tauopathy. However, AV-1451 accumulation was not associated with psychometric performance in our cohort.

C9orf72 is required for proper macrophage and microglial function in mice.

Expansions of a hexanucleotide repeat (GGGGCC) in the noncoding region of the C9orf72 gene are the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. Decreased expression of C9orf72 is seen in expansion carriers, suggesting that loss of function may play a role in disease. We found that two independent mouse lines lacking the C9orf72 ortholog (3110043O21Rik) in all tissues developed normally and aged without motor neuron disease. Instead, C9orf72 null mice developed progressive splenomegaly and lymphadenopathy with accumulation of engorged macrophage-like cells. C9orf72 expression was highest in myeloid cells, and the loss of C9orf72 led to lysosomal accumulation and altered immune responses in macrophages and microglia, with age-related neuroinflammation similar to C9orf72 ALS but not sporadic ALS human patient tissue. Thus, C9orf72 is required for the normal function of myeloid cells, and altered microglial function may contribute to neurodegeneration in C9orf72 expansion carriers.

On the relation between the activation energy for electron attachment reactions and the size of their thermal rate coefficients.

Rate coefficients k(T) for dissociative electron attachment (DEA) to molecules in many cases exhibit a more or less strong rise with increasing temperature T (the electron temperature T(e) and the molecular temperature T(G) are assumed to be in thermal equilibrium, i.e., T = T(e) = T(G)). This rise is frequently modeled by the Arrhenius equation k(T) = k(A) exp[-E(a)∕(k(B)T)], and an activation energy E(a) is deduced from fits to the experimental data k(T). This behavior reflects the presence of an energy barrier for the anion on its path to the dissociated products. In a recent paper [J. Kopyra, J. Wnorowska, M. Forys, and I. Szamrej, Int. J. Mass Spectrom. 268, 60 (2007)] it was suggested that the size of the rate coefficients for DEA reactions at room temperature exhibits an exponential dependence on the activation energy, i.e., k(E(a); T ≈ 300 K) = k(1) exp[-E(a)∕E(0)]. More recent experimental data for molecules with high barriers [T. M. Miller, J. F. Friedman, L. C. Schaffer, and A. A. Viggiano, J. Chem. Phys. 131, 084302 (2009)] are compatible with such a correlation. We investigate the validity and the possible origin of this dependence by analyzing the results of R-matrix calculations for temperature-dependent rate coefficients of exothermic DEA processes with intermediate barrier toward dissociation. These include results for model systems with systematically varied barrier height as well as results of molecule-specific calculations for CH(3)Cl, CH(3)Br, CF(3)Cl, and CH(2)Cl(2) (activation energies above 0.2 eV) involving appropriate molecular parameters. A comparison of the experimental and theoretical results for the considered class of molecules (halogenated alkanes) supports the idea that the exponential dependence of k(T = 300 K) on the activation energy reflects a general phenomenon associated with Franck-Condon factors for getting from the initial neutral vibrational levels to the dissociating final anion state in a direct DEA process. Cases are discussed for which the proposed relation does not apply.

[Composition and levels of phospholipids in plasma from patients with exacerbation of chronic gastritis].

Blood plasma phospholipids from 45 patients with chronic gastritis and 20 healthy volunteers have been investigated. Chronic gastritis exacerbation is accompanied by essential change in the spectrum and concentration of the phospholipids. The absolute concentrations of the phospholipids and such fractions as phosphatidylcholine, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine and sphingomyelin significally increased while concentrations of phosphatidilinositol and lysophosphatidylserine decreased. The period of the remission of the disease was accompanied by a certain normalization of the level and spectrum phospholipids and literature data suggest that phospholipids and their particular fractions are involved into pathogenesis of the inflammatory process in gastric mucosa and in the process of reparative regeneration aswell.

HETC radiation transport code development for cosmic ray shielding applications in space.

In order to facilitate three-dimensional analyses of space radiation shielding scenarios for future space missions, the Monte Carlo radiation transport code HETC is being extended to include transport of energetic heavy ions, such as are found in the galactic cosmic ray spectrum in space. Recently, an event generator capable of providing nuclear interaction data for use in HETC was developed and incorporated into the code. The event generator predicts the interaction product yields and production angles and energies using nuclear models and Monte Carlo techniques. Testing and validation of the extended transport code has begun. In this work, the current status of code modifications, which enable energetic heavy ions and their nuclear reaction products to be transported through thick shielding, are described. Also, initial results of code testing against available laboratory beam data for energetic heavy ions interacting in thick targets are presented.

Correlating phenotype and genotype in the periodic paralyses.

BACKGROUND: Periodic paralyses and paramyotonia congenita are rare disorders causing disabling weakness and myotonia. Mutations in sodium, calcium, and potassium channels have been recognized as causing disease. OBJECTIVE: To analyze the clinical phenotype of patients with and without discernible genotype and to identify other mutations in ion channel genes associated with disease. METHODS: The authors have reviewed clinical data in patients with a diagnosis of hypokalemic periodic paralysis (56 kindreds, 71 patients), hyperkalemic periodic paralysis (47 kindreds, 99 patients), and paramyotonia congenita (24 kindreds, 56 patients). For those patients without one of the classically known mutations, the authors analyzed the entire coding region of the SCN4A, KCNE3, and KCNJ2 genes and portions of the coding region of the CACNA1S gene in order to identify new mutations. RESULTS: Mutations were identified in approximately two thirds of kindreds with periodic paralysis or paramyotonia congenita. The authors found differences between the disorders and between those with and without identified mutations in terms of age at onset, frequency of attacks, duration of attacks, fixed proximal weakness, precipitants of attacks, myotonia, electrophysiologic studies, serum potassium levels, muscle biopsy, response to potassium administration, and response to treatment with acetazolamide. CONCLUSIONS: Hypokalemic periodic paralysis, hyperkalemic periodic paralysis, and paramyotonia congenita may be distinguished based on clinical data. This series of 226 patients (127 kindreds) confirms some clinical features of this disorder with notable exceptions: In this series, patients without mutations had a less typical clinical presentation including an older age at onset, no changes in diet as a precipitant, and absence of vacuolar myopathy on muscle biopsy.

First observation of four-body breakup in electron recombination: C2D+5.

We report the first observation of four-body breakup in electron dissociative recombination of a molecular ion: C2D+5. In an ion storage ring experiment, the branching ratio for the process C2D+5 + e(-)-->C2D2 + D + D + D was determined to be 13%. This means that three covalent chemical bonds are broken as a result of the action of a single electron. This is the first time a four-body breakup of chemical bonds has been observed in a low-energy binary reaction.

Double-differential heavy-ion production cross sections.

Current computational tools used for space or accelerator shielding studies transport energetic heavy ions either using a one-dimensional straight-ahead approximation or by dissociating the nuclei into protons and neutrons and then performing neutron and proton transport using Monte Carlo techniques. Although the heavy secondary particles generally travel close to the beam direction, a proper treatment of the light ions produced in these reactions requires that double-differential cross sections should be utilised. Unfortunately, no fundamental nuclear model capable of serving as an event generator to provide these cross sections for all ions and energies of interest exists currently. Herein, we present a model for producing double-differential heavy-ion production cross sections that uses heavy-ion fragmentation yields produced by the NUCFRG2 fragmentation code coupled with a model of energy degradation in nucleus-nucleus collisions and systematics of momentum distributions to provide energy and angular dependences of the heavy-ion production.

Double-differential light-ion production cross sections.

Codes used for space radiation shielding studies typically transport light-ions from the incident cosmic ray spectrum using either a one-dimensional straight-ahead approximation or in three dimensions, often without accounting for their breakup. Light-ions are also produced by breakup of energetic heavy ions in target materials. Herein, we present cross section models that can be used to predict double-differential light-ion production cross sections. Deuteron breakup is based on a parameterization of the total reaction cross sections. Alpha fragmentation utilizes a parameterization based on an abrasion-ablation model that has been augmented by experimental data. Neutron and proton production is modelled with a quantum mechanical abrasion-ablation-coalescence model. This same model is also used to predict cross sections for triton and 3He breakup. Finally, energies and emission angles for particles other than nucleons are specified using a model of energy degradation in nucleus-nucleus collisions and systematics of momentum distributions.

Particle formation by ion nucleation in the upper troposphere and lower stratosphere.

Unexpectedly high concentrations of ultrafine particles were observed over a wide range of latitudes in the upper troposphere and lower stratosphere. Particle number concentrations and size distributions simulated by a numerical model of ion-induced nucleation, constrained by measured thermodynamic data and observed atmospheric key species, were consistent with the observations. These findings indicate that, at typical upper troposphere and lower stratosphere conditions, particles are formed by this nucleation process and grow to measurable sizes with sufficient sun exposure and low preexisting aerosol surface area. Ion-induced nucleation is thus a globally important source of aerosol particles, potentially affecting cloud formation and radiative transfer.

Effectiveness of Agency for Health Care Policy and Research clinical practice guideline and patient education methods for pregnant smokers in medicaid maternity care.

OBJECTIVE: The purposes of this study were (1) to determine the extent to which tobacco exposure assessment and new patient education methods, derived from a meta-analysis and the Agency for Health Care Policy and Research guideline recommendations, could be provided routinely by trained Medicaid maternity care staff members and (2) to document the behavioral impact of these interventions among pregnant smokers. STUDY DESIGN: After 265 pregnant smokers were assigned at their first visit to an experimental group (n = 139) or a control group (n = 126), they received standardized risk information and were advised to quit smoking. The experimental group also received evidence-based patient education methods, including the videocassette Commit to Quit During and After Pregnancy, the publication A Pregnant Woman's Guide to Quit Smoking, and a brief counseling session. Self-report and saliva cotinine assessments of tobacco exposure were performed at baseline and at the end of pregnancy. RESULTS: A significantly higher percentage of patients quit smoking in the experimental group (17.3%) than in the control group (8.8%). CONCLUSIONS: The application of principles of organizational development and quality improvement at the management and clinical practice levels and the delivery of evidence-based health education methods by trained prenatal care providers significantly increased smoking cessation rates among pregnant Medicaid recipients.

Enhancement of clinicians' diagnostic reasoning by computer-based consultation: a multisite study of 2 systems.

CONTEXT: Computer-based diagnostic decision support systems (DSSs) were developed to improve health care quality by providing accurate, useful, and timely diagnostic information to clinicians. However, most studies have emphasized the accuracy of the computer system alone, without placing clinicians in the role of direct users. OBJECTIVE: To explore the extent to which consultations with DSSs improve clinicians' diagnostic hypotheses in a set of diagnostically challenging cases. DESIGN: Partially randomized controlled trial conducted in a laboratory setting, using a prospective balanced experimental design in 1995-1998. SETTING: Three academic medical centers, none of which were involved in the development of the DSSs. PARTICIPANTS: A total of 216 physicians: 72 at each site, including 24 internal medicine faculty members, 24 senior residents, and 24 fourth-year medical students. One physician's data were lost to analysis. INTERVENTION: Two DSSs, ILIAD (version 4.2) and Quick Medical Reference (QMR; version 3.7.1), were used by participants for diagnostic evaluation of a total of 36 cases based on actual patients. After training, each subject evaluated 9 of the 36 cases, first without and then using a DSS, and suggested an ordered list of diagnostic hypotheses after each evaluation. MAIN OUTCOME MEASURE: Diagnostic accuracy, measured as the presence of the correct diagnosis on the hypothesis list and also using a derived diagnostic quality score, before and after consultation with the DSSs. RESULTS: Correct diagnoses appeared in subjects' hypothesis lists for 39.5% of cases prior to consultation and 45.4% of cases after consultation. Subjects' mean diagnostic quality scores increased from 5.7 (95% confidence interval [CI], 5.5-5.9) to 6.1 (95% CI, 5.9-6.3) (effect size: Cohen d = 0.32; 95% CI, 0.23-0.41; P<.001). Larger increases (P = .048) were observed for students than for residents and faculty. Effect size varied significantly (P<.02) by DSS (Cohen d = 0.20; 95% CI, 0.08-0.32 for ILIAD vs Cohen d = 0.45; 95% CI, 0.31-0.59 for QMR). CONCLUSIONS: Our study supports the idea that "hands-on" use of diagnostic DSSs can influence diagnostic reasoning of clinicians. The larger effect for students suggests a possible educational role for these systems.

The effect of professional education on the knowledge and attitudes of physical therapist and occupational therapist students about acquired immunodeficiency syndrome.

BACKGROUND AND PURPOSE: Anxiety and fear about caring for people with acquired immunodeficiency syndrome (AIDS) are concerns expressed by students in health care professions. This study was designed to evaluate the influence of education offered to physical therapist (PT) and occupational therapist (OT) students on their knowledge, attitudes, and willingness to provide services to people with AIDS. SUBJECTS AND METHODS: Twenty-six undergraduate PT students and 23 undergraduate OT students completed a questionnaire at the beginning of their professional education program, following a 5-hour AIDS education seminar, and shortly before their graduation. The questionnaire consisted of 3 subscales designed to evaluate the respondents' knowledge, attitudes, and willingness to treat people with AIDS. RESULTS: At the time of graduation, the students in both disciplines showed improvement in knowledge about AIDS (14.3% for PT students and 13.8% for OT students) and more positive attitudes toward people with AIDS (7.4% for PT students and 5% for OT students). In both disciplines, the students' willingness to provide services for people with AIDS remained unchanged following the AIDS education seminar and at the end of the professional education program. CONCLUSION AND DISCUSSION: The professional education offered to the cohort of students in this study appeared to be beneficial in improving their knowledge and attitudes toward people with AIDS, but it did not affect their willingness to work with this patient group.

Bax deletion further orders the cell death pathway in cerebellar granule cells and suggests a caspase-independent pathway to cell death.

Dissociated cerebellar granule cells maintained in medium containing 25 mM potassium undergo an apoptotic death when switched to medium with 5 mM potassium. Granule cells from mice in which Bax, a proapoptotic Bcl-2 family member, had been deleted, did not undergo apoptosis in 5 mM potassium, yet did undergo an excitotoxic cell death in response to stimulation with 30 or 100 microM NMDA. Within 2 h after switching to 5 mM K+, both wild-type and Bax-deficient granule cells decreased glucose uptake to <20% of control. Protein synthesis also decreased rapidly in both wild-type and Bax-deficient granule cells to 50% of control within 12 h after switching to 5 mM potassium. Both wild-type and Bax -/- neurons increased mRNA levels of c-jun, and caspase 3 (CPP32) and increased phosphorylation of the transactivation domain of c-Jun after K+ deprivation. Wild-type granule cells in 5 mM K+ increased cleavage of DEVD-aminomethylcoumarin (DEVD-AMC), a fluorogenic substrate for caspases 2, 3, and 7; in contrast, Bax-deficient granule cells did not cleave DEVD-AMC. These results place BAX downstream of metabolic changes, changes in mRNA levels, and increased phosphorylation of c-Jun, yet upstream of the activation of caspases and indicate that BAX is required for apoptotic, but not excitotoxic, cell death. In wild-type cells, Boc-Asp-FMK and ZVAD-FMK, general inhibitors of caspases, blocked cleavage of DEVD-AMC and blocked the increase in TdT-mediated dUTP nick end labeling (TUNEL) positivity. However, these inhibitors had only a marginal effect on preventing cell death, suggesting a caspase-independent death pathway downstream of BAX in cerebellar granule cells.

Inhibition of phosphatidylinositol 3-kinase activity blocks depolarization- and insulin-like growth factor I-mediated survival of cerebellar granule cells.

Depolarizing concentrations of potassium promote the survival of many neuronal cell types including cerebellar granule cells. To begin to understand the intracellular mediators of neuronal survival, we have tested whether the survival-promoting effect of potassium depolarization on cerebellar granule cells is dependent on either mitogen-activated protein (MAP) kinase or phosphatidylinositol 3-kinase (PI-3-K) activity. In 7-day cerebellar granule cell cultures, potassium depolarization activated both MAP kinase and PI-3-K. Preventing the activation of MAP kinase with the MEK1 inhibitor PD98059 did not affect potassium saving. In contrast, the survival-promoting effect of 25 mM potassium was negated by the addition of 30 microM LY 294002 or 1 microM wortmannin, two distinct inhibitors of PI-3-K. The cell death induced by PI-3-K inhibition was indistinguishable from the cell death caused by potassium deprivation; LY 294002-induced death included nuclear condensation, was blocked by cycloheximide, and had the same time course as potassium deprivation-induced cell death. Cerebellar granule cells can also be maintained in serum-free medium containing either 100 ng/ml insulin-like growth factor I (IGF-I) or 800 microM cAMP. PI-3-K inhibition completely blocked the survival-promoting activity of IGF-I, but had no effect on cAMP-mediated survival. These data indicate that the survival-promoting effects of depolarization and IGF-I, but not cAMP, require PI-3-K activity.

Metabolic and genetic analyses of apoptosis in potassium/serum-deprived rat cerebellar granule cells.

Cerebellar granule cells maintained in medium containing serum and 25 mM potassium undergo an apoptotic death within 96 hr when switched to serum-free medium with 5 mM potassium. Because large numbers of apparently homogeneous neurons can be obtained, this represents a potentially useful model of neuronal programmed cell death (PCD). Analysis of the time course and extent of death after removal of either serum or K+ alone demonstrated that a fast-dying (T(1/2) = 4 hr) population (20%) responded to serum deprivation, whereas a slow-dying (T(1/2) = 25 hr) population (80%) died in response to K+ deprivation. Taking advantage of the complete death after removing both K+ and serum, changes in metabolic events and mRNA levels were analyzed in this model. Glucose uptake, protein synthesis, and RNA synthesis fell to <35% of control by 9 hr after potassium/serum deprivation, a time when 85% of the cells were still viable. The pattern of the fall in these metabolic parameters was similar to that reported for trophic factor-deprived sympathetic neurons. Most mRNAs decreased markedly after K+/serum deprivation. Levels of c-jun mRNA increased fivefold in potassium/serum-deprived granule cells; c-jun is required for cell death of sympathetic neurons. mRNA levels of cyclin D1, c-myb, collagenase, and transin remained relatively constant in potassium/serum-deprived granule cells. These data demonstrate the existence of two populations of granule cells with respect to cell death and define common metabolic and genetic events involved in neuronal PCD.

Academic performance is not a viable determinant of physical therapy students' burnout.

This study investigated the proposed link between students' academic performance and burnout. We found no significant correlations between students' cumulative grade point average and ratings of emotional exhaustion, depersonalization, and personal accomplishment.