cssi/SciSciGPT-SciSciNet huggingface dataset:
Single-step fabrication of electrochemical flow cells utilizing multi-material 3D printing

Abstract Here we present methodology for fabricating electrochemical flow cells with embedded carbon-composite electrodes in a single step using simultaneous 3D printing of insulating poly(lactic acid) (PLA) and a commercially available graphene–PLA composite. This work is significant because it is the first demonstration that devices capable of fluid handling and electrochemical sensing can be produced in a single fabrication step using inexpensive equipment. We demonstrate the broad utility of this approach using a channel-flow configuration as an exemplary system for hydrodynamic electrochemistry. Unmodified devices were characterized using hydrodynamic electrochemistry, and behave according to the well-established Levich equation. We also characterized the fabrication reproducibility and found that the devices were within 3% RSD. The 3D-printed sensors we employed were subsequently modified by electroplating Au and used under flowing conditions to detect catechol, whose oxidation requires two electrons and two protons and is thus more challenging to analyze than the outer-sphere FcCH2OH. We envision these results will pave the way for the development of highly customized micro-total analysis systems that include embedded electrochemical sensors for a variety of redox-active analytes.
Population Pharmacokinetics of Clofarabine as Part of Pretransplantation Conditioning in Pediatric Subjects before Hematopoietic Cell Transplantation.

ABSTRACT The primary objective of this work was to characterize the pharmacokinetics (PK) of systemic clofarabine (clo-fara) in pediatric allogeneic hematopoietic cell transplantation (HCT) recipients receiving either nucleoside monotherapy or a dual nucleoside analog preparative regimen. Fifty-one children (median age, 4.9 years; range, .25 to 14.9 years) undergoing allogeneic HCT for a variety of malignant and nonmalignant disorders underwent PK assessment. Plasma samples were collected over the 4 to 5 days of clo-fara treatment and quantified for clo-fara, using a validated liquid chromatography/tandem mass spectrometry assay. Nonlinear mixed-effects modeling was used to develop the population PK model, including identification of covariates that influenced drug disposition. In agreement with previously published models, a 2-compartment PK model with first-order elimination best described the PK of clo-fara. Final parameter estimates for clo-fara were consistent with previous reports and were as follows: clearance (CL), 23 L/h/15 kg; volume of the central compartment, 42 L/15 kg; volume of peripheral compartment, 47 L/15 kg; and intercompartmental CL, 9.8 L/h/15 kg. Unexplained variability was acceptable at 33%, and the additive residual error (reflective of the assay) was estimated to be 0.36 ng/mL. Patient-specific factors significantly impacting clo-fara CL included actual body weight and age. The covariate model was able to estimate clo-fara CL with good precision in children spanning a wide age range from infancy to early adulthood and demonstrates the need for variable dosing in children of different ages. For example, the dose required for a 6-month and 1-year old was approximately 43% and 17% lower, respectively, than the typical 40 mg/m2dose to achieve the median AUC0-24of 1.04 mg·h/L in the study population. Despite the known renal elimination of clo-fara, no significant clinical parameters for renal function were retained in the final model (P> .05). Coadministration of fludarabine with clo-fara did not alter the CL of clo-fara (P> .05). These results will help inform individualized dosing strategies for clo-fara to improve clinical outcomes and limit drug-related adverse events in children undergoing HCT.
What’s Wrong with On-the-Fly Partial Order Reduction

Partial order reduction and on-the-fly model checking are well-known approaches for improving model checking performance. The two optimizations interact in subtle ways, so care must be taken when using them in combination. A standard algorithm combining the two optimizations, published over twenty years ago, has been widely studied and deployed in popular model checking tools. Yet the algorithm is incorrect. Counterexamples were discovered using the Alloy analyzer. A fix for a restricted class of property automata is proposed.
Design of a Wind Speed Sensor Based on MEMS

This paper presents the design, fabrication and test results of a 2D hot-air speed sensor based on the micro-electro-mechanical technology. The sensor uses constant power operation mode and uses the method of thermal temperature difference to measure the wind speed and direction. The sensor uses the micro-electro-mechanical peeling technology to fabricate both heating resistance and temperature-measuring resistance on glass substrates. Thermal isolation and high sensitivity are achieved by depending on the processing technology.
Political respect for nature

Political respect for nature is an important part of cultivating a more emancipatory and ecologically sustainable politics. As a political principle, it can supplement respect for persons with inst...
Suppression of pyramidal neuron G protein-gated inwardly rectifying K+ channel signaling impairs prelimbic cortical function and underlies stress-induced deficits in cognitive flexibility in male, but not female, mice.

Imbalance in prefrontal cortical (PFC) pyramidal neuron excitation:inhibition is thought to underlie symptomologies shared across stress-related disorders and neuropsychiatric disease, including dysregulation of emotion and cognitive function. G protein-gated inwardly rectifying K+ (GIRK/Kir3) channels mediate excitability of medial PFC pyramidal neurons, however, the functional role of these channels in mPFC-dependent regulation of affect, cognition, and cortical dynamics is unknown. We used a viral-cre approach in male and female mice harboring a "floxed" version of the kcnj3 (Girk1) gene, to disrupt GIRK1-containing channel expression in pyramidal neurons within the prelimbic cortex (PrL). In males, loss of pyramidal GIRK1-dependent signaling differentially impacted measures of affect and impaired working memory and cognitive flexibility. Unexpectedly, ablation of PrL GIRK1-dependent signaling did not impact affect or cognition in female mice. Additional studies used a model of chronic unpredictable stress (CUS) to determine the impact on PrL GIRK-dependent signaling and cognitive function. CUS exposure in male mice produced deficits in cognition that paralleled a reduction in PrL pyramidal GIRK-dependent signaling akin to viral approaches whereas CUS exposure in female mice did not alter cognitive flexibility performance. Stress-induced behavioral deficits in male mice were rescued by systemic injection of a novel, GIRK1-selective agonist, ML297. In conclusion, GIRK1-dependent signaling in male mice, but not females, is critical for maintaining optimal PrL function and behavioral control. Disruption of this inhibition may underlie stress-related dysfunction of the PrL and represent a therapeutic target for treating stress-induced deficits in affect regulation and impaired cognition that reduce quality of life.
Hemoadsorption for management of patients on veno-venous ECMO support for severe COVID-19 acute respiratory distress syndrome.

Background and aim Patients with severe coronavirus disease 2019 (COVID-19) develop a profound cytokine-mediated pro-inflammatory response. This study reports outcomes in 10 patients with COVID-19 supported on veno-venous extracorporeal membrane oxygenation (VV-ECMO) who were selected for the emergency use of a hemoadsorption column integrated in the ECMO circuit. Materials and methods Pre and posttreatment, clinical data, and inflammatory markers were assessed to determine the safety and feasibility of using this system and to evaluate the clinical effect. Results During hemoadsorption, median levels of interleukin (IL)-2R, IL-6, and IL-10 decreased by 54%, 86%, and 64%, respectively. Reductions in other markers were observed for lactate dehydrogenase (-49%), ferritin (-46%), d-dimer (-7%), C-reactive protein (-55%), procalcitonin (-76%), and lactate (-44%). Vasoactive-inotrope scores decreased significantly over the treatment interval (-80%). The median hospital length of stay was 53 days (36-85) and at 90-days post cannulation, survival was 90% which was similar to a group of patients without the use of hemoadsorption. Conclusions Addition of hemoadsorption to VV-ECMO in patients with severe COVID-19 is feasible and reduces measured cytokine levels. However, in this small series, the precise impact on the overall clinical course and survival benefit still remains unknown.
Education and the World of Objective Knowledge

Let me start first with a statement of a misconception of science that I have used when arguing against the reductionist dogma that everything including biology and even all of conscious experience is reducible to physics and chemistry. In the past I have raised the question : What then is physics and chemistry? It will be readily agreed that it is not the text books of physics and chemistry or the scientific publications as such or the institutions and laboratories of these sciences. It is not just even the physicists and chemists per se. So I went on further to assert that, in the ultimate analysis, physics and chemistry are just the thoughts and ideas of physicists and chemists in their efforts to understand and so to explain the manifestations of the natural world as directly observed or as observed in experimental procedures. This assertion led to the further statement that physics and chemistry had in that way been reduced to a subjective category, and that the reductionist dogma as stated above therefore could not be regarded as supporting a materialistic philosophy.
BRST Current Algebra Derivation of the Higher Cocycles

Two topics of much current research in mathematical physics are the BRST quantization of gauge theories and the understanding of anomalies in gauge theories. While a large part of the stimulus for this research is the revival of interest in string theory, much remains to be learned about gauge theories. Here, we shall present a new extension of BRST symmetry in the form of a local symmetry and demonstrate that this leads to an understanding, in the context of a field theory, of the higher cocycles that have been mathematically associated with chiral anomalies.1 We obtain, furthermore, a unification of many approaches to the chiral anomaly. Namely, once the BRST symmetry is gauged, we can examine BRST current algebra. Anomalies and the cocyles associated with them are consequences of this current algebra. Moreover, the BRST charge derived from the BRST current is not nilpotent.
Finding the Pearls When the World Is Your Oyster: Case and Project Selection in Clinic Design

Clinical legal education is distinguishable from the rest of the law school curriculum and the extracurricular activities available to law students because it places students directly into the role of a lawyer engaged in real-world practice. Clinical programs are often defined by the cases and projects — the pearls at the heart of the experiential learning experience — that comprise their dockets. Finding the right cases and projects that meet a range of goals remains a perennial challenge in clinic design. In the context of international human rights clinics, the world is your oyster, and that challenge is magnified. This Article identifies a set of core lawyering competencies — skills and values — essential to successful practice in a globalized world, and argues that those competencies should drive the peda-gogical goals that serve as the criteria against which to assess the value of the clinical teaching pearls. Building on prior scholarship by international human rights clinical law faculty, and analyzing case studies from the Transnational Legal Clinic at the University of Pennsylvania Law School, this Article then examines the benefits and challenges for students engaged in a range of individual and project-based human rights representations. This Article concludes with the provocative and important question: How can we prevent the risk of harm to the often unfamiliar and fragile ecosystems into which we dive in search of our teaching pearls?
Instruction-driven wake-up mechanisms for Snoopy TAP controller

The issue of handling multiple embedded 1149.1-compliant TAP controllers (as part of embedded cores) in a 1149.1-compliant IC, has received significant attention, in recent times. The Hierarchical TAP (HTAP) architecture, previously developed by this author, provides a systematic solution to this problem, by allowing multiple embedded TAPs to share one set of 1149.1-specified test pins on the IC. The HTAP architecture requires an augmented version of the 1149.1-specified TAP-designated the Snoopy TAP (SNTAP)-in the top-level of the design hierarchy, that handles 1149.1-specified TAP functions for the non-TAP'ed parts of the IC, and performs arbitration of the shared IC pins between multiple embedded TAPs. Previously, a mechanism was developed to wake up the SNTAP using a simple string of constant values (0 or 1) on the TMS input. Two new mechanisms to wake up SNTAP by means of an instruction scanned in, are described in this report. Both mechanisms allow embedded 1149.1-compliant TAP's to be used in a hierarchical design, as is, without any hardware modification. Moreover, both mechanisms enable reuse of the test program from the embedded TAP'ed cores, without any modification. The instructions to wake up the SNTAP-to switch access to different TAP-ed cores-can be simply inserted in-between the test programs for the embedded cores.
Maternal-Fetal Transfer of Ascorbic Acid in the Guinea Pig

Placental transfer of ascorbic acid was studied in the guinea pig using an in situ placental perfusion technique. A total of 14 animals were studied during their last week of gestation. The fetus was shown to have higher plasma total ascorbic acid (TAA) levels than the mother prior to maternal ascorbic acid infusion. However, following maternal venous infusion this normal gradient for TAA (fetal greater than maternal) disappeared and a new gradient was established (maternal greater than fetal). As maternal plasma TAA levels rose, the level of TAA in the placental perfusate steadily increased to a maximal concentration of approximately 0.85 mg/dl. This occurred when maternal plasma TAA levels were above 3.0 mg/dl. These results demonstrate that a saturable or carrier-mediated transport mechanism is involved in the placental transfer of ascorbic acid in this species. The apparent Vmax (the maximal transport rate by the saturable system per gram placenta) and the Kt (or one-half saturation concentration) were estimated to be 8.3 nmoles/minute and 0.12 mM, respectively. Additional results are presented to suggest that dehydroascorbic acid (DHA) may be the predominant form of the vitamin crossing the guinea pig placenta.
Systematic analysis of design and stratification for phase III trials in first‐line advanced non‐small cell lung cancer

Background A recent study reviewed phase III trials of first-line advanced non-small cell lung cancer (NSCLC) conducted from 1981 to 2010, and provided trends in the study outcome. However, such trials have never been analyzed in detail for design and stratification factors. Methods Phase III studies of systemic treatment for first-line advanced or metastatic NSCLC published in English literature between 1981 and 2010 were identified. Characteristics, including sample size, number of trials, region, rate of meeting accrual goal, primary endpoint, type of phase III, interim analysis, allocation method, and stratification factors, were determined for each decade. Results A total of 162 studies met the criteria. The number of studies and sample size increased over the three decades. The primary endpoint was reported more frequently in recent decades, and non-overall survival endpoints were chosen in European and Asian studies. Interim analysis was conducted more commonly during the 2000s. Allocation method was rarely reported throughout the three decades. The number of stratification factors increased significantly from one in 1980s to three in 2000s. Performance status, stage, and institution were most frequently selected, and at least one of the three factors was used in most of the studies in the 2000s. However, there are many other stratification factors that were used infrequently. Conclusions Despite Consolidated Standards of Reporting Trials guidelines, allocation method has rarely been reported. The choice of stratification factor remains inconsistent across studies.
Robust stability and stabilization of time delay systems in real parameter space

This note provides some results about robust stability and stabilization of time delay systems in real parameter space via Lyapunov and Riccati equation approaches.
CVD‐Silizium als Korrosionsschutz für die Beschaufelung von stationären Gasturbinen

Zur Erhohung der Korrosionsbestandigkeit von hochwarmfesten Nickellegierungen, die als Schaufelmaterialien fur stationare Gasturbinen Verwendung finden, wurden diese Materialien nach einem CVD-Verfahren (Chemical Vapor Deposition) mit Silizium beschichtet. Mit Hilfe von Mikrosonden- und Rontgenuntersuchungen wurden die gebildeten Silizide identifiziert. Beschichtete Proben wurden einer Reihe von spezifisch auf den Einsatz in stationaren Gasturbinen ausgerichteten Prufverfahren ausgesetzt. Dabei zeigte sich, das Silizidschichten auf Nickellegierungen auserordentlich bestandig gegen Hochtemperaturkorrosion sind und eine wesentliche Erhohung der Standzeiten von silizierten Gasturbinenschaufeln erwarten lassen. Chemical Vapor Deposited silicon as a corrosion protection for the blades of stationary gas turbines In order to improve the corrosion resistance of heat resistant nickel alloys used as the blade materials for stationary gas turbines these materials were given a silicon coating by a chemical vapour deposition process. With the aid of microprobe and X-ray investigations the silicides formed were investigated. Coated specimens were exposed to a series of tests specifically oriented toward the intended use in stationary gas turbines. It turned out, that the silicide coatings on nickel base alloys are exceptionally stable to high temperature corrosion, so that a considerable increase of the useful life of silicided gas turbine blades may be expected.
Flexibility in Strategic Decision Making: Informational and Ideological Perspectives

Adaptation is a crucial challenge for organizations, and an important theme in the strategy and organization theory literature. We still have much to learn, however, about the strategic processes by which adaptation is achieved. In this paper we focus on a basic element in the adaptation process, i.e. flexibility within the strategic decision-making process. We concentrate on strategic decisions because these choices are the most important adaptations the firm makes. We suggest that the core of all organizational adaptation is a decision-making process. Unless the decision-making process itself is flexible, it is unlikely the organization can be flexible enough to adapt. We derive hypotheses concerning the factors that lead to flexibility (versus rigidity) from both information processing and ideological perspectives, and test them in a study involving 57 strategic decisions in 25 companies. Our results identify three contextual factors related to both perspectives -- including competitive threat, slack and uncertainty -- that are helpful in understanding flexibility in strategy decision making. While managers appear to be more flexible when decisions are uncertain, we found that in the very conditions where managers need the most flexibility (high competitive threat and low slack), they are least likely to be flexible.
Explaining the heritability of an ecologically significant trait in terms of individual quantitative trait loci

Most natural populations display substantial genetic variation in behaviour, morphology, physiology, life history and the susceptibility to disease. A major challenge is to determine the contributions of individual loci to variation in complex traits. Quantitative trait locus (QTL) mapping has identified genomic regions affecting ecologically significant traits of many species. In nearly all cases, however, the importance of these QTLs to population variation remains unclear. In this paper, we apply a novel experimental method to parse the genetic variance of floral traits of the annual plant Mimulus guttatus into contributions of individual QTLs. We first use QTL-mapping to identify nine loci and then conduct a population-based breeding experiment to estimate VQ, the genetic variance attributable to each QTL. We find that three QTLs with moderate effects explain up to one-third of the genetic variance in the natural population. Variation at these loci is probably maintained by some form of balancing selection. Notably, the largest effect QTLs were relatively minor in their contribution to heritability.
Saturation of band tail states in a-Si:H

Abstract The excitation intensity dependence of the intrinsic photoluminescence (PL) vs. temperature for sputtered and glow discharge hydrogenated amorphous silicon is reported. We find that a transition from linearity to sub-linearity in the excitation intensity dependence of the PL intensity is consistent the onset of photocarrier band tail saturation.
Closed-form direct displacement analysis of a 6-6 Stewart platform

Abstract The direct displacement analysis of a class of 6-6 Stewart platforms is considered here. Even though this is not the most general possible configuration, it has many of the advantages of the general configuration, and includes several useful arrangements that are of practical significance. The analysis leads to closed-form, decoupled polynomial equations (a quartic and two quadratics) of overall degree equal to sixteen. Since the largest degree polynomial is a quartic, no iterative schemes are required during the analysis. Finally, it is shown that only a few of the sixteen configurations have to be solved for algebraically, and the other configurations can be generated by geometric reasoning.
Localization and function of Kinesin-5-like proteins during assembly and maintenance of mitotic spindles in Silvetia compressa

Background Kinesin-5 (Eg-5) motor proteins are essential for maintenance of spindle bipolarity in animals. The roles of Kinesin-5 proteins in other systems, such as Arabidopsis, Dictyostelium, and sea urchin are more varied. We are studying Kinesin-5-like proteins during early development in the brown alga Silvetia compressa. Previously, this motor was shown to be needed to assemble a bipolar spindle, similar to animals. This report builds on those findings by investigating the localization of the motor and probing its function in spindle maintenance.
An exploratory study of the effects of frequency and duration of messaging on impression development in computer-mediated communication

This experimental study indicates that computer-mediated communication (CMC) is a viable mode of social-emotion-oriented communication. In this study, the effects of frequency and duration of messa...
QUEUEING MODELS WITH DELAYED STATE-DEPENDENT SERVICE TIMES

Abstract In a state-dependent queueing model, the service rate is adjusted at both epochs of customer arrivals and departures. In certain situations, frequently changing service rates is neither economical nor practical. This paper studies an alternative queueing model in which the server changes its service rates, or service types, only at the beginning of service. The model is analyzed by using an embedded Markov chain technique. The probability generating functions for the steady-state queue length are determined for both queueing models with and without vacations. Two particular models are considered in detail. One is the delayed M/M/ c queue and the other is the delayed M/G i /1 queue with c service types. The performance measures such as the mean queue length and additional waiting time are calculated and compared with regular queues.
Heterogeneous oxidation kinetics of organic biomass burning aerosol surrogates by O3, NO2, N2O5, and NO3.

The reactive uptake coefficients (γ) of O3, NO2, N2O5, and NO3 by levoglucosan, abietic acid, nitroguaiacol, and an atmospherically relevant mixture of those species serving as surrogates for biomass burning aerosol have been determined employing a chemical ionization mass spectrometer coupled to a rotating-wall flow-tube reactor. γ of O3, NO2, N2O5, and NO3 in the presence of O2 are in the range of 1−8 × 10−5, <10−6−5 × 10−5, 4−6 × 10−5, and 1−26 × 10−3, respectively, for the investigated organic substrates. Within experimental uncertainties the uptake of NO3 was not sensitive to relative humidity levels of 30 and 60%. NO3 uptake experiments involving substrates of levoglucosan, abietic acid, and the mixture exhibit an initial strong uptake of NO3 followed by NO3 gas-phase recovery as a function of NO3 exposure. In contrast, the uptake of NO3 by nitroguaiacol continuously proceeds at the same efficiency for investigated NO3 exposures. The derived oxidative power, i.e. the product of γ and atmospheric oxidant concentration, for applied oxidants is similar or significantly larger in magnitude than for OH, emphasizing the potential importance of these oxidants for particle oxidation. Estimated atmospheric lifetimes for the topmost organic layer with respect to O3, NO2, N2O5, and NO3 oxidation for typical polluted conditions range between 1–112 min, indicating the potential for significant chemical transformation during atmospheric transport. The contact angles determined prior to, and after heterogeneous oxidation by NO3, representative of 50 ppt for 1 day, do not decrease and thus do not indicate a significant increase in hygroscopicity with potential impacts on water uptake and cloud formation processes.
Uncertainty and risk in aircraft structures: current status and recommended directions

Widespread interest in uncertainty quantification methods for airframe design and certification is driven by the desire to realise acquisition and operational cost savings through increased reliance on analysis; the goal being to design and produce more robust airframes. General sources of uncertainty are described, that complicate airframe design and testing, thereby contributing to the high cost of airframes. Recent applications of uncertainty quantification are reviewed with the objective of highlighting promising research and transition applications. Finally, several challenges and needs are explored to suggest future steps that must be completed to enable practical application of uncertainty quantification in airframe design and certification.
Glomerular β-galactosidase expression following transduction with microsphere-adenoviral complexes

Glomerular β-galactosidase expression following transduction with microsphere-adenoviral complexes. The aortic injection of adenoviral-microsphere complexes is a useful technique for in vivo gene transfer (transduction) to the glomerulus. In this approach, the appearance of the foreign transprotein in the glomerulus may result from glomerular cell gene transfer and local synthesis or hepatic cell transduction followed by synthesis, secretion, and deposition in the glomerulus. We postulated that glomerular expression of the foreign transgene was the result of glomerular cell transduction. To test this question, male SD rats underwent aortic injections with adenovirus containing the LacZ expression cassette [expressing β-galactosidase (βgal)] coupled to 16 μm diameter microspheres. After 48 hours, histologic staining confirmed glomerular expression of the βgal transprotein and reverse transcription in situ polymerase chain reaction demonstrated the presence of the βgal transgene in the glomerulus. In addition, hepatic expression of the βgal transprotein was minimal and substantially less than that observed in the glomeruli. These data support the contention that adenoviral-microsphere complexes result in glomerular cell transduction with the desired transgene, followed by local transprotein synthesis. This approach may prove useful for facilitating glomerular gene transfer in the development of gene therapy for glomerulonephritis.
Lumen Morphogenesis in C. elegans Requires the Membrane-Cytoskeleton Linker erm-1

Abstract Epithelial tubes are basic building blocks of complex organs, but their architectural requirements are not well understood. Here we show that erm-1 is a unique C. elegans ortholog of the ERM family of cytoskeleton-membrane linkers, with an essential role in lumen morphogenesis. ERM-1 localizes to the luminal membranes of those tubular organ epithelia which lack stabilization by cuticle. RNA interference (RNAi), a germline deletion, and overexpression of erm-1 cause cystic luminal phenotypes in these epithelia. Confocal and ultrastructural analyses indicate that erm-1 functions directly in apical membrane morphogenesis, rather than in epithelial polarity and junction assembly as has been previously proposed for ERMs. We also show that act-5 /cytoplasmic actin and sma-1 /β-H-spectrin are required for lumen formation and functionally interact with erm-1 . Our findings suggest that there are common structural constraints on the architecture of diverse organ lumina.
Sorting of carboxypeptidase E to the regulated secretory pathway requires interaction of its transmembrane domain with lipid rafts.

Carboxypeptidase E (CPE) functions as a regulated secretory pathway sorting receptor for several prohormones, including pro-opiomelanocortin (POMC), proenkephalin and proinsulin. The association of CPE with lipid rafts in the trans -Golgi network and secretory granule membranes is necessary for its sorting receptor function. We now provide evidence that a domain within the C-terminal 25 residues of CPE functions as a signal for both raft association and the sorting of CPE to the regulated secretory pathway. A fusion protein containing the extracellular domain of the human interleukin-2 receptor Tac (N-Tac) and the C-terminal 25 amino acids of CPE was transfected into Neuro2A cells. This fusion protein floated in sucrose density gradients, indicating raft association, and co-localized with chromogranin A (CGA), a secretory granule marker. To define further a minimum sequence required for raft association and sorting, deletion mutants of CPE that lacked the C-terminal four or 15 residues (CPE-Delta4 and CPE-Delta15 respectively) were transfected into a clone of CPE-deficient Neuro2A cells. In contrast with full-length CPE, neither CPE-Delta4 nor CPE-Delta15 floated in sucrose density gradients. The sorting of both CPE-Delta4 and CPE-Delta15 to the regulated secretory pathway was impaired, as indicated by significantly increased basal secretion and a lack of response to stimulation. Additionally, there was a significant decrease in the co-localization of mutant CPE immunofluorescence with CGA when compared with full-length CPE. Finally, the sorting of the prohormone POMC to the regulated pathway was impaired in cells transfected with either CPE-Delta4 or CPE-Delta15. We conclude that the sorting of CPE to the regulated secretory pathway in endocrine cells is mediated by lipid rafts, and that the C-terminal four residues of CPE, i.e. Thr(431)-Leu-Asn-Phe(434), are required for raft association and sorting.
Abstract LB-251: Manipulation of macrophage phenotype enhances the apoptotic response to chemotherapy in mesothelioma

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Macrophages within the tumor microenvironment (TME) may contribute to tumor chemoresistance. We have found that macrophages constitute a significant percentage of the infiltrating leukocytes in mesothelioma, a highly chemoresistant tumor. We asked whether reprogramming of macrophage phenotype from a Th2 (pro-tumor) to a Th1 (anti-tumor) phenotype would alter the mesothelioma chemoresponsiveness. To address this, we used 3D organotypic models of mesothelioma growth: 1) multicellular spheroids, with mesothelioma cells grown alone or co-cultured with macrophages derived from peripheral blood monocytes, and 2) primary tumor fragment spheroids derived from small fragments of freshly isolated human mesothelioma. Macrophages were incubated with Th1 (LPS & IFNγ) or Th2 (IL-4 &IL-13) cytokines prior to co-culture with mesothelioma spheroids, followed by exposure to standard-of-care chemotherapy, cisplatin plus pemetrexed. Gene expression was evaluated in macrophages to affirm Th1-type (TNF, IL-12, IFNα) versus Th2-type (CD206, IL10) programming. In both spheroid approaches, Th1-type macrophage programming significantly increased the apoptotic response of mesothelioma cells to chemotherapy. Moreover, when CSF1R signaling in macrophages was inhibited by incubation with a small molecular weight kinase antagonist, GW2850, chemoresponsiveness was significantly increased as evidenced by increased presence of apoptotic tumor cells in spheroids. Enhanced chemoresponsiveness of mesothelioma tumor cells was dependent on presence of macrophages because incubation of tumor cells with Th1-type cytokines or GW2850 in the absence of macrophages was without effect. We conclude that manipulating the TME may be a promising therapeutic approach in mesothelioma. This work was supported by a DOD Mesothelioma Program grant PR80717 to Broaddus and Coussens and a T32 Training Grant to Battula and Blakely. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-251. doi:1538-7445.AM2012-LB-251
Enhanced osteoclastogenesis causes osteopenia in twisted gastrulation-deficient mice through increased BMP signaling.

The uncoupling of osteoblastic and osteoclastic activity is central to disorders such as osteoporosis, osteolytic malignancies, and periodontitis. Numerous studies have shown explicit functions for bone morphogenetic proteins (BMPs) in skeletogenesis. Their signaling activity has been shown in various contexts to be regulated by extracellular proteins, including Twisted gastrulation (TWSG1). However, experimental paradigms determining the effects of BMP regulators on bone remodeling are limited. In this study, we assessed the role of TWSG1 in postnatal bone homeostasis. Twsg1-deficient (Twsg1−/−) mice developed osteopenia that could not be explained by defective osteoblast function, because mineral apposition rate and differentiation markers were not significantly different compared with wildtype (WT) mice. Instead, we discovered a striking enhancement of osteoclastogenesis in Twsg1−/− mice, leading to increased bone resorption with resultant osteopenia. Enhanced osteoclastogenesis in Twsg1−/− mice was caused by increased cell fusion, differentiation, and function of osteoclasts. Furthermore, RANKL-mediated osteoclastogenesis and phosphorylated Smad1/5/8 levels were enhanced when WT osteoclasts were treated with recombinant BMP2, suggesting direct regulation of osteoclast differentiation by BMPs. Increase in detectable levels of phosphorylated Smad 1/5/8 was noted in osteoclasts from Twsg1−/− mice compared with WT mice. Furthermore, the enhanced osteoclastogenesis in Twsg1−/− mice was reversed in vitro in a dose-dependent manner with exposure to Noggin, a BMP antagonist, strongly suggesting that the enhanced osteoclastogenesis in Twsg1 mutants is attributable to increased BMP signaling. Thus, we present a novel and previously uncharacterized role for TWSG1 in inhibiting osteoclastogenesis through regulation of BMP activity.
Purification of the two complementary subunits of ribonucleotide reductase from calf thymus

Summary Subunits M1 and M2 of calf thymus ribonucleotide reductase have been purified to homogeneity. Each runs as a single band in SDS-polyacrylamide gel electrophoresis with polypeptide molecular weights of 84,000 for M1 and 58,000 for M2. Additional evidence for homogeneity was obtained using high performance liquid gel chromatography. The native M1 subunit is predominantly monomeric under experimental conditions while the M2 subunit is oligomeric. The M2 subunit has a uv-visible absorption spectrum with a major peak at 404 nm but this spectrum is different from that of the B2 subunit of E. coli ribonucleotide reductase.
Characterization of hematopoietic cells arising on the textured surface of left ventricular assist devices

Background. Textured biomaterial surfaces in implantable left ventricular assist devices induce development of a nonthrombotic neointimal surface and allow elimination of anticoagulation therapy in device recipients. Characterization of the hematopoietic cells formed within the neointimal surfaces of these devices will contribute to our understanding of this unique neointima. Methods. The blood-contacting surface of seven ThermoCardiosystems left ventricular assist devices was removed, washed with phosphate-buffered saline solution, and digested with 0.1% collagenase for 15 to 20 minutes. The hematopoietic cells released from the explants were isolated and analyzed by flow cytometry and immunohistochemical staining. Results. More than 80% ± 6% of hematopoietic cells isolated in this fashion are of myelomonocytic origin and express CD14, CD15, and CD33 surface molecules. Four percent of cells express the CD34 surface marker, which suggests that the neointima is colonized by pluripotent hematopoietic stem cells. Continuous culture of these hematopoietic cells in the presence of the cytokines interleukin-3, c-kit ligand, granulocyte colony—stimulating factor, and granulocyte/macrophage colony—stimulating factor resulted in tenfold expansion by day 7 and 25-fold expansion by day 14. Conclusions. Pluripotent hematopoietic cells with a high proliferative capacity colonize textured surfaces of left ventricular assist devices and may contribute to the development of a biologically nonthrombogenic neointima.
Complex-valued analysis and visualization of fMRI data for event-related and block-design paradigms

Independent Component Analysis (ICA) has been noted to be promising for the study of functional magnetic resonance imaging (fMRI) data also in its native complex-valued form. In this paper, we demonstrate the first successful application of group ICA to complex-valued fMRI data of an event-related paradigm. We show that networks associated with event-related responses as well as intrinsic fluctuations of hemodymamic activity can be extracted for data collected during an auditory oddball paradigm. The intrinsic networks are of particular interest due to their potential to study cognitive function and mental illness, including schizophrenia. More importantly, we show that analysis of fMRI data in its complex form can increase the sensitivity and specificity in the detection of activated brain regions both for event-related and block design paradigms when compared to magnitude-only applications. In addition, we introduce a novel fMRI phase-based visualization (FPV) technique to identify activated voxels such that the complex nature of the data is fully taken into account.
An automated accelerometry system for gait analysis

Abstract An automated system for assessing gait as a motor function is described. The system is composed of electronic footswitch devices, a triaxial accelerometer, and amplifier-power supply interface integrated with a PDP-12 computer. Data acquisition, data analysis, and validation procedures are discussed. Preliminary results of a study involving tibio-femoral knee implants demonstrates the applicability of the automated system to evaluation of motor performance during gait.
Population ecology and niche seeking in the development of gay and lesbian rights groups

Abstract This paper explores the formation of groups within the framework of the population ecology literature. Specifically, I argue that newly formed groups’ target membership bases and policy agendas will become more focused as the density of interest groups in a population increases, though groups will not necessarily continue this focused trend once the density of the population has reached its upper limits. To test this theory, I utilize Nownes’ [Nownes, A. J. (2004). The population ecology of interest group formation: Mobilizing for gay and lesbian rights in the United States, 1950–1998. British Journal of Political Science, 34, 49–67] dataset on the population ecology of gay and lesbian rights groups and a typology of groups that Bosso [Bosso, C. J. (2005). Environment, Inc.: From Grassroots to Beltway. Lawrence: University Press of Kansas] used in explaining the focus of environmental rights groups. I find that as the population of gay and lesbian groups became more dense, the groups that formed tended to narrow down their potential membership populations. However, the results were different for a newly formed group's policy agenda. While niche seeking occurred during the rapid growth of groups in the 1970s and the early 1980s, groups that formed after growth leveled off in the mid-1980s balanced their agendas between being too broad and too narrow. This important finding indicates that niche seeking does not always occur in very dense interest populations.
Some behavioral consequences of H-2:Qa:Tla polymorphism☆☆☆

The conjunction of a penetrating idea and a novel observation is an experimentalist’s delight. At about the time that Lewis Thomas was writing on histocompatibility genes and olfaction it was brought to my attention that the social behavior of certain mice of our colonies at Sloan-Kettering Institute suggested that they could sense differences in the H-2 haplotypes of their companions. In short, males and females of dissimilar H-2 type appeared to associate with one another in preference to partners of the same H-2 type. Since the mice in question were congenic, differing only at the H-2 region of chromosome 17, the inference was that H-2 or linked genes are concerned in a sensory communication system among mice. The upshot of succeeding studies was the demonstration of spontaneous mating preferences which, at least among congenic laboratory mice, generally favor H-2 disparity (l-4). Subsequent proof that chemosensory recognition is involved in the identification of H-2 types among mice came from the introduction of a Y-maze testing system which has been exploited with gratifying effect at the Monell Chemical Senses Center in Philadelphia (5). In this method, mice are trained to distinguish the arms of a Y-maze scented by odors of alternative sets of H-2 congenic mice. Training consists in prior withholding of water and provision of a drop of water each time the trainee mouse selects whichever of the two scents is assigned for reinforcement. It was found that urine is a potent source of H-2 distinctive odors (6), and thus that olfaction, which is a function of primary sensory neurons whose bodies are situated in the olfactory mucosa, sufficiently explains the discrimination of H-2 types. We have asked whether the genetic constitution of the odor phenotype is restricted to H-2 as defined by its K and D gene poles. This is not so. Recombinant congenic strains which reduce the genetic difference to the Qa:Tla region adjacent to H-2 can be distinguished in the Y maze, and disparity restricted to the region of H-2K alone is also distinguishable (7). Thus the odor phenotype corresponding to a given H-2
Abstract 2033: Transcription factors that regulate MUC16 expression in ovarian cancer

Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL CA125 antigen is found to be elevated in 85% of patients diagnosed with advanced epithelial ovarian cancer. CA125 has a minimal role as a screening modality. The gene which encodes for the CA125 glycoprotein, MUC16, has been previously identified and sequenced. MUC16 has a molecular size of 22,000 bps and consists of cytoplasmic domain, transmembrane region and 9 external domain tandem repeats. The large molecular weight of this gene has made it difficult to study and track. N terminal signal peptide and transcription factor or factors that triggers MUC16 in ovarian cancer is not well demonstrated. The proximal 114 amino acids of MUC16 can transform NIH/3T3 cells and increase soft agar formation and matrigel invasion. We have designed several constructs from the region about 1000 nucleotides upstream and 300 nucleotides downstream of the MUC16 start Methionine with secretory pMetridia Luciferase Reporter vector (Clonetech, CA). Four constructs of varying lengths had the MUC16 UTR TSS site whereas two did not. Putative binding sites for transcription factors like ER, STAT1, STAT3, CRE-BP1 (ATF2) and NFKB are also present. These constructs were then transfected individually into CA125 negative ovarian cell line, SK-Ov-3, and also into CA125 positive ovarian cell lines, SK-Ov-8 and CAOV3. All cell lines were stably selected with G418 and were used within 10 passages. Western blots confirm the presence of ERalpha, STAT1 and STAT3. Stably transfected cell lines were cultured in low serum conditions with or without 17β-Estradiol or IL-1β, were used to see the effect of transcription. Clinical CA125 levels were also documented for each construct. Secretory pMetridia Luciferase signals were detected in the supernant of the culture cells. There is no difference between untreated or 17β-Estradiol or IL-1β treated cells. However, gel retardation studies with OVCAR3 and SKOV8 wild type cell lines suggested CREB/ATF (CREBZF) transcription factor involvement and confirmatory reporter gene studies are planned. Currently, we are assessing the putative transcription factor binding sites within the 1300 nucleotides region from which we derived the constructs. We conclude that these studies will lead us to better understand the biology of MUC16 gene expression and give us an insight about the role of transcription factors. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2033. doi:10.1158/1538-7445.AM2011-2033
Isolation of acetylpeptide from enzymic digests of TMV-protein.

Abstract N-acetylseryltyrosine was isolated from the chyromotryptic and peptic digests of TMV-protein. The structure of the acetylpeptide purified by ion-exchange chromatography, was studied by the FDNB method, hydrazinolysis and the use of carboxypeptidase. For the characterization of the acyl group in the peptide, hydrazinolysis was also used. The location of the acyl peptide in the protein was discussed.
Articulating Meaning: Reconceptions of the Meaning Process, Fantasy/Reality, and Identity in Leisure Activities

This article reexamines the complexities of the reception of meaning by audience members who experience media texts. It argues for the articulation model of meaning, which asserts that meaning is a momentaty event, denies that meaning is a transfer process, and characterizes media experiences as intertwined and blurred fantasy and reality experiences that serve to confirm identity for the receiver. The articulation model resolves key issues in the cultural studies debates. Framed by an articulation analysis, an empirical study of adolescent romance readers demonstrates the articulation model's power to yield additional and “corrective” insights into meaning.