Role of Group X Secretory Phospholipase A\u3csub\u3e2\u3c/sub\u3e in Murine Adipocytes

The secretory phospholipase A2 (sPLA2) family is a group of enzymes that catalyze the hydrolysis of glycerophospholipids at the sn-2 position, generating free fatty acids and lysophospholipids. The sPLA2 family has been implicated in various physiological and pathological activities. Eleven sPLA2’s have been identified in mammals, and the function of each isoform likely reflects its tissue distribution and substrate specificity. Studies in vitro indicate that Group X (GX) sPLA2 potently releases arachidonic acid (AA) and lysophosphatidylcholine from mammalian cell membranes. Interestingly, some of the biological effects mediated by GX sPLA2 in vitro are independent of its catalytic activity. Despite a wealth of in vitro data, the in vivo function of GX sPLA2 still remains to be elucidated. In order to define the function of GX sPLA2 in vivo, our laboratory recently generated C57BL/6 mice with targeted deletion of GX sPLA2 (GX-/- mice). When fed a normal rodent diet, GX-/- mice gained significantly more weight and had increased adiposity compared to GX+/+ mice, which was not attributable to alterations in food consumption or energy expenditure. When treated with adipogenic stimuli ex vivo, stromal vascular cells isolated from adipose tissue of GX-/- mice accumulated significantly more (20%) triglyceride compared to cells from GX+/+ mice. Conversely, overexpression of GX sPLA2, but not catalytically inactive GX sPLA2, resulted in a significant 50% reduction in triglyceride accumulation in OP9 adipocytes. The induction of adipogenic genes, including PPAR-γ, SREBP-1c, SCD-1 and FAS was also significantly blunted by 50-80% in OP9 cells overexpressing GX sPLA2. Activation of the liver X receptor (LXR), a nuclear receptor known to upregulate adipogenic gene expression, was suppressed in 3T3-L1 and OP9 cells when GX sPLA2 was overexpressed. Thus, hydrolytic products generated by GX sPLA2 negatively regulate adipogenesis, possibly by suppressing LXR activation

Lithics in the West: Using Lithic Analysis to Solve Archeological Problems in Western North America

Lithics in the West seeks to link the rich archaeological lithic data base from the western United States with some of the contemporary theoretical and analytical approaches used in global settings in stone tool and debitage analysis today. The book highlights the role that lithic analysis (in all its forms) plays in solving research problems in the prehistory of western North America. The book covers important archaeological sites and projects in Montana, Wyoming, Idaho, Colorado, and Washington. Contributors include William Andrefsky, Jr., Robert Kelly, Nicole Waguespack, Pei-Lin Yu, Doug MacDonald, Robert Brunswig, Scott Carpenter, Jackie Cook, David Diggs, Philip Fisher, Katie Harris, Brian Ostahowski, Mary Prasciunas, Ken Reid, and Todd Surovell.https://scholarworks.umt.edu/umpress-oabooks/1000/thumbnail.jp

Wheels within wheels: an examination of the nature of psychological explanation via a theoretically oriented history of some mechanical models

The aim of this thesis is to ask, and attempt to answer, some pertinent questions about that type of psychological explanation which proceeds by simulation, or model building. The method chosen is a detailed examination of some models, mostly 18th and 19th century mechanical ones, together with a theoretically motivated discussion of the relations between these models and the development of psychological theories contemporary with them. Two types of model, formal and intimate, are distinguished, both by their aetiology and by the way they are used by working scientists, and several examples of each type are subjected to scrutiny, as are the intentions of their modellers in building or adopting them. Four main foci of interest emerge: the history of experimental psychology (the myth that experimental psychology was born circa 1870 is exploded); the sociology of science (the impact of developing technology on psychological theory, via the proffering of models, is clearly demonstrated); the philosophy of psychology (issues such as the nature of explanation and the problem of representation are dis¬ cussed); and, last but not least, theoretical psychology (the value of work in cognitive simulation, and of some work in Artificial Intelligence, is stressed and, partly, explained)

Novel insights into the function and regulation of group X secretory phospholipase A\u3csub\u3e2\u3c/sub\u3e

Group X secretory phospholipase A2 (GX sPLA2) hydrolyzes membrane phospholipids producing free fatty acids and lysophospholipids. Previous studies from our lab suggest that mice with targeted deletion of GX sPLA2 (GX KO) have increased age-related weight gain due to an increase in overall adiposity. Paradoxically, this increased adiposity is associated with improved age-related glucose intolerance. GX KO mice also demonstrate a reduced inflammatory response to lipopolysaccharide injection. In vitro studies indicate this phenotype may be attributable to blunted macrophage mediated inflammatory responses. Given the role of macrophages in promoting adipose tissue (AT) inflammation and metabolic dysfunction in response to diet-induced obesity, we hypothesized that GX KO mice would be protected from the obesity related metabolic derangements associated with overfeeding. Unexpectedly, GX KO mice were only partially protected from high fat (HFD) diet-induced glucose intolerance and showed no improvement in HFD-induced insulin resistance. Moreover, GX KO mice were not protected against HFD-induced AT inflammation. GX sPLA2 is produced as a proenzyme (pro-GX sPLA2), and propeptide cleavage is required for enzymatic activity. Furin-like proprotein convertases (PCs) have recently been implicated in the proteolytic activation of pro-GX sPLA2; however the identity of individual PCs involved is unclear. Previous findings from our lab have shown that GX sPLA2 is expressed in the adrenals where it regulates glucocorticoid production. GX KO mice have increased plasma corticosterone levels under both basal and ACTH-induced stress conditions. However, how GX sPLA2 is regulated in the adrenals is still uncertain. We hypothesized that PCs may be involved in the proteolytic activation of pro-GX sPLA2 in the adrenals. Here we report the novel findings that the PCs, furin and PCSK6, proteolytically activate pro-GX sPLA2 in Y1 adrenal cells. Furthermore, we demonstrate that PC dependent processing of pro-GX sPLA2 is necessary for GX sPLA2 dependent suppression of steroidogenesis. Finally, we provide evidence that pro-GX sPLA2 processing by PCs is enhanced in response to adrenocorticotropic hormone (ACTH), suggesting a novel mechanism for negatively regulating adrenal steroidogenesis. Cumulatively, these studies provide valuable insight into the function and regulation of GX sPLA2

Processing and application of polyolefin plastics

This thesis describes the manufacturing, processing and application of polyolefin thermoplastics. Polyolefins have become one of the most important kinds of plastics. Polyolefin resin represents about one third of all plastics sold and its application for films and containers dominate the packaging industry. The objective of this thesis is to present the various processing methods and application of polyolefin thermoplastics. Polyolefin is produced by several different processes. Conventional low density polyethylene is made by polymerizing ethylene at high pressure and high temperatures while high density polyethylene is polymerized at relatively low pressure and low temperatures. Generally all polyolefin possess excellent electrical properties, excellent resistance to water and moisture and good resistance to chemicals. They are translucent, light weight, tough and flexible material. This combination of properties makes them suitable for film, blow, and injection molded parts, flexible sheet and extruded profiles, and tubing

New Approaches in Automation and Robotics

The book New Approaches in Automation and Robotics offers in 22 chapters a collection of recent developments in automation, robotics as well as control theory. It is dedicated to researchers in science and industry, students, and practicing engineers, who wish to update and enhance their knowledge on modern methods and innovative applications. The authors and editor of this book wish to motivate people, especially under-graduate students, to get involved with the interesting field of robotics and mechatronics. We hope that the ideas and concepts presented in this book are useful for your own work and could contribute to problem solving in similar applications as well. It is clear, however, that the wide area of automation and robotics can only be highlighted at several spots but not completely covered by a single book

Charting PARP-1 dependent mechanisms for DNA double-strand break resection

L'intégrité de l'ADN génomique humain est maintenue par des systèmes de réparation de l'ADN qui protègent les cellules des dommages causés par des agents environnementaux ou des lésions spontanées de l'ADN. Chaque cellule peut subir jusqu'à 10⁵ lésions par jour, y compris les cassures double-brin de l'ADN (CDB). La poly(ADPribosyl)ation (PARylation) est l'un des premiers événements de signalisation moléculaire survenant aux CDBs. Il est catalysé par les poly(ADP-ribose)polymérases (PARP) qui sont directement activées par ces lésions d'ADN. Le fait de ne pas générer de poly(ADP)ribosyl (pADPr) en réponse à des dommages à l'ADN par une inhibition chimique ou par l'absence de PARP-1 augmente la sensibilité cellulaire au stress génotoxique, indiquant que la pADPr elle-même est une molécule clé de signalisation des dommages à l'ADN. L'inhibition de l'enzyme de signalisation des dommages à l'ADN, la poly(ADP-ribose) polymérase-1 (PARP-1) est l'une des nouvelles thérapies les plus prometteuses contre le cancer. Les inhibiteurs de PARP sensibilisent les cellules cancéreuses aux agents endommageant l'ADN et tuent efficacement les cellules cancéreuses du sein, des ovaires et du pancréas déficientes en BRCA1 (Breast Cancer gene 1) et BRCA2 (Breast Cancer gene 2), ce qui suggère que les cellules déficientes en réparation des CDBs sont extrêmement sensibles à l'inhibition de PARP. Pourtant, les mécanismes sous-jacents à cette létalité synthétique entre le déficit de réparation du CDB et l'inhibition de PARP restent mal définis. Il y a un débat considérable sur le mécanisme par lequel l'inhibition de PARP tue les cellules déficientes en réparation de l'ADN, et le plein potentiel des inhibiteurs de PARP dans le traitement du cancer ne peut être obtenu que par une compréhension claire des voies de réponse aux dommages de l'ADN (DDR) aux CDB et comment ils sont affectés par les inhibiteurs de PARP. L'objectif général de ma thèse est d'étudier le rôle de PARP-1 dans la réparation DSB et d'identifier les interacteurs de PARP-1 qui jouent également un rôle dans ce processus. Les cellules eucaryotes réparent les CDBs par deux voies principales, la jonction d'extrémité non homologue (NHEJ) et la recombinaison homologue (HR). La HR est initiée par la liaison des CDBs par BRCA1 et le complexe MRE11-RAD50-NBS1 et des nucléases EXO1/DNA2 pour générer de l'ADN simple-brin, qui est ensuite utilisé par la recombinase RAD51 et le complexe BRCA1-PALB2-BRCA2. Une question clé dans notre domaine concerne les facteurs critiques pour réguler le choix de la voie CDB. HR est initiée à partir d'extrémités DSB hautement résectées, tandis que dans le NHEJ, la résection est empêchée par des facteurs de réparation clés incluant RIF1 et 53BP1. En utilisant des cellules déficientes en PARP-1, nous avons observé que deux inhibiteurs de la résection de l'ADN et des régulateurs de choix de voie, RIF1 et 53BP1, la formation de foyers induits par des dommages à l'ADN sont fortement altérés. Cela confirme notre hypothèse selon laquelle PARP-1 participe à la réparation du DSB en influençant la résection de l'ADN. Afin de mieux comprendre le mécanisme de résection et le rôle que PARP-1 y joue, nous avons identifié d'autres protéines qui interagissent avec PARP-1 et modulent ce processus. Pour ce faire, nous avons utilisé des données sur les protéines de liaison au pADPr générées à la fois dans notre laboratoire et celui de notre collaborateur Ted Dawson de Johns Hopkins. Les candidats sélectionnés à partir de ces listes ont été criblés pour identifier une seule cible qui démontrerait un phénotype similaire à la perte de PARP-1. Deux cibles initiales ont été explorées et finalement une seule protéine à doigt de zinc a été choisie comme cible principale. Nous devons relever la fonction de ce doigt de zinc en HR, dans l'espoir qu'il permettra de découvrir davantage les mécanismes de PARP-1 en résection. En résumé, cette thèse élucide le rôle de PARP-1 dans la résection de l'ADN et identifie une protéine à doigt de zinc non étudiée auparavant qui interagit avec PARP-1 et partage une fonction similaire à PARP-1 dans la résection de l'ADN.The integrity of human genomic DNA is maintained by DNA repair systems that will protect cells from damage by environmental agents or spontaneous DNA lesions. Each cell can experience up to 10⁵ lesions daily, including DNA double-strand breaks (DSB)s. Poly(ADP-ribosyl)ation (PARylation) is one of the earliest molecular signalling events occurring at DNA DSBs. It is catalysed by poly(ADP-ribose) polymerases (PARPs) that are directly activated by those DNA lesions. Failure to generate pADPr in response to DNA damage by either chemical inhibition or absence of PARP-1 increases the cellular sensitivity to genotoxic stress, indicating that pADPr itself is a key DNA damage signalling molecule. Inhibition of the DNA damage signalling enzyme poly(ADP-ribose) polymerase-1 (PARP-1) is among the most promising new therapies in cancer. PARP inhibitors sensitize cancer cells to DNA damaging agents and efficiently kill BRCA1- and BRCA2-deficient breast, ovarian and pancreatic cancer cells, suggesting that cells deficient in DSB repair are exquisitely sensitive to PARP inhibition. Yet, the mechanisms underlying this synthetic lethality between DSB repair deficiency and PARP inhibition remain poorly defined. There is considerable debate about the mechanism through which PARP inhibition kills DNA repair-deficient cells, and the full benefit of PARP inhibitors in cancer therapy can only be achieved by a clear understanding of the DNA damage response (DDR) pathways to DSBs and how these are affected by PARP inhibitors. The overall aim of my PhD is to investigate the role of PARP-1 in DSB repair and identify interactors of PARP-1 which also play a role in this process. Eukaryotic cells repair DSBs by two major pathways, non-homologous end-joining (NHEJ) and homologous recombination (HR). HR is initiated by the binding of DSB by BRCA1 and the end resection of the DSB by MRE11 (and the associated NBS1, RAD50, CtIP, and EXO1) to generate single-stranded DNA, which is further processed by RAD51 and BRCA1-PALB2-BRCA2. A key question in our field regards which factors are critical for regulating the DSB pathway choice. HR is initiated from highly resected DSB ends, whereas in NHEJ, resection is prevented by key repair factors that include RIF1 and 53BP1. Using PARP-1-deficient cells, we have observed that two inhibitors of DNA resection and regulators of pathway choice, RIF1 and 53BP1, are strongly impaired in forming DNA damage-induced foci. This supports our hypothesis that PARP-1 participates in DSB repair by influencing DNA resection. In order to further understand the mechanism of resection and the role that PARP-1 plays in it we also aim to identify other proteins which interact with PARP-1 and modulate this process. To accomplish this, we made use of data on PAR binding proteins generated both in our lab and that of our collaborator Ted Dawson. The candidates selected from these lists were screened to identify a single target that would demonstrate a similar phenotype to PARP-1 loss. Two initial targets were further explored and finally a single zinc finger protein was selected as our primary target. We aim to characterize the function of this zinc finger in HR, in the hopes that it will further uncover the mechanisms of PARP-1 in resection. In summary this thesis elucidates the role of PARP-1 in DNA resection and identifies a previously unstudied zinc finger protein which interacts with PARP-1 and shares a similar function to PARP-1 in DNA resection

Study of lipid bilayer behaviour modified by substrate interactions

Biological membranes rarely exist as free-floating structures but are often confined and supported by various cellular assemblies such as the cytoskeleton and the extracellular matrix. It has already been shown that biological and polymeric substrates can modulate the morphology and response to various stimuli of supported lipid bilayers significantly. The interaction between such structures and the membrane are obviously important yet remain poorly understood even in minimal or synthetic systems. The work of this thesis utilises a variety of fluorescence microscopy and atomic force microscopy (AFM) techniques to investigate the behaviour and structure of supported lipid bilayers, in particular how interfacial features of their support substrate influence and modulate their morphology and biophysical properties. First, surface modification of polydimethylsiloxane is systematically explored, in particular how the interfacial properties of such a polymer substrate can be modified to create fully and partially plasma-treated interfaces that stably support lipid bilayers. Lipid patch formation on such substrates is then investigated, revealing that the membrane undergoes significant morphological reorganisation after vesicle fusion has completed forming a lipid patch. The underlying mechanisms can be altered by substrate interactions following different pathways for fully and partially plasma-treated PDMS substrates. Furthermore, partially plasma-treated substrates are demonstrated to be capable of specifically depleting cholesterol from supported lipid membranes, while stably supporting the other remaining phospholipid species. Studies of cholesterol depletion of lipid patches possessing liquid-ordered and disordered domains reveal a disruption in domains structure, with the partitioning of fluorescent dyes into regions from which they were previously excluded. This structure perturbation was found to be reversible upon the reinsertion of cholesterol into the bilayer. Many of the discussed mechanisms are only observed in the presence of a substrate, emphasising the importance of substrate interactions in both functional biomembranes and the development of supported membrane technologie

Signal processing for malware analysis

This Project is an experimental analysis of Android malware through images. The analysis is based on classifying the malware into families or differentiating between goodware and malware. This analysis has been done considering two approaches. These two approaches have a common starting point, which is the transformation of Android applications into PNG images. After this conversion, the first approach was subtracting each image from the testing set with the images of the training set, in order to establish which unknown malware belongs to a specific family or to distinguish between goodware and malware. Although the accuracy was higher than the one defined in the requirements, this approach was a time consuming task, so we consider another approach to reduce the time and get the same or better accuracy. The second approach was extracting features from all the images and then using a machine learning classifier to get a precise differentiation. After this second approach, the resulting time for 100,000 samples was less than 4 hours and the accuracy 83.04%, which fulfill the requirements specified. To perform the analysis, we have used two heterogeneous datasets. The Malgenome dataset which contains 49 kinds of malware Android applications (49 malware families). It was used to perform the measurements and the different tests. The M0droid dataset, which contains goodware and malware Android applications. It was used to corroborate the previous analysis.Este proyecto es un análisis experimental de aplicaciones de Android mediante imágenes. Este análisis se basa en clasificar las imágenes en familias o en diferenciarlas entre goodware o malware. Para ello, se han considerado dos enfoques. Estas dos aproximaciones tienen como punto en común la transformación de las aplicaciones de Android en imágenes de tipo PNG. Después de este proceso de transformación a imágenes, la primera aproximación se basó en restar cada imagen perteneciente al grupo de pruebas con las imágenes del grupo de entrenamiento, de esta forma se pudo saber la familia a la que pertenecía cada malware desconocido o distinguir entre aplicaciones goodware y malware. Sin embargo, a pesar de que la precisión de acierto era más alta que la definida en los requisitos, este enfoque era una tarea que consumía mucho tiempo, así que consideramos otra aproximación para reducir el tiempo y conseguir una precisión parecida o mejor que la anterior. Este segundo enfoque fue extraer las características de las imágenes para después usar un clasificador y así obtener una diferenciación precisa. Con esta segunda aproximación, conseguimos un tiempo total menor a las 4 horas para 100000 muestras con una precisión del 83.04%, cumpliendo y superando de esta forma los requisitos que habían sido especificados. Este análisis se ha llevado a cabo usando dos sets de datos heterogéneos. Uno de ellos fue el perteneciente a un proyecto llamado Malgenome, éste contiene 49 tipos de familias de malware en Android. El set de datos de Malgenome se usó para realizar los diferentes ensayos o pruebas y sobre el que se realizaron las medidas de tiempo y precisión. El set de datos de M0droid se usó para corroborar el análisis previo y así establecer una clasificación final.Ingeniería Informátic

Proceedings of the Advanced Hadron Facility accelerator design workshop, February 20--25, 1989

The International Workshop on Hadron Facility Technology was held February 20--25, 1989, at the Study Center at Los Alamos National Laboratory. This volume (second of two) included papers on computer controls, polarized beam, rf, magnet and power supplies, experimental areas, and instabilities. Participants included groups from AHF, Brookhaven National Laboratory, European Hadron Facility, Fermilab, and the Moscow Meson Factory. The workshop was well attended by members of the Los Alamos staff. The interchange of information and the opportunity by criticism by peers was important to all who attended