Abstract only
The CB1 cannabinoid receptor is the most abundantly expressed GPCR in the CNS and modulates many diverse neuronal functions related to synaptic plasticity and neuroprotection. The ...cellular actions of CB1 ligands on neuroprotection are poorly understood. A recent demonstration that the cannabinoid receptor interacting protein (CRIP1a) had the ability to switch CB1 neuroprotection from an agonist‐ to antagonist‐driven mechanism is intriguing (Stauffer et al., 2011 Neuroscience Letters). To better understand the role of CRIP1a in cytoprotection, we developed neuronal cell models to over‐express or knock‐down CRIP1a in order to identify changes in CB1‐mediated signal transduction and gene transcription. Data from On‐cell‐Western experiments indicate that over‐expression of CRIP1a significantly reduces the levels of phosphoERK, a downstream kinase and transcript factor activated by CB1. Additional studies focusing on phospholipase C and NFkappaB pathways demonstrated that modulation of CRIP1a produces alterations in the mRNA levels of c‐Fos, CCAAT/Enhancer binding protein (Cebpb), NF‐kappaB pathway and genes involved in cell survival and neuroprotection. These experiments suggest that CRIP1a may modulate CB1 signaling and transcription of genes that are important in stress and cell survival pathways.
Supported by NIH grants T32‐DA007246, R01‐DA003690, R21‐ DA025321, K01‐DA024763.
3D printing has seen an explosion of interest and growth in recent years, especially within the biomedical space. Prized for its efficiency, ability to produce complex geometries, and facile material ...processing, additive manufacturing is rapidly being used to create medical devices ranging from orthopedic implants to tissue scaffolds. However, 3D printing is currently limited to a select few material choices, especially when one considers soft tissue replacement or augmentation. To this end, my research focuses on developing material systems that are simultaneously 1) 3D printable, 2) biocompatible, and 3) mechanically robust with properties appropriate for soft-tissue replacement or augmentation applications. Two systems were developed toward this goal: an interpenetrating network (IPN) hydrogel consisting of covalently crosslinked poly (ethylene glycol) diacrylate (PEGDA) and ionically crosslinked brown sodium alginate, and semi-crystalline thiol-ene photopolymers containing spiroacetal molecules in the polymer main-chain backbone. In addition to successfully being incorporated into existing 3D printing systems (extrusion-deposition for the PEGDA-alginate hydrogel and digital light processing for the thiol-ene polymers) both systems exhibited biocompatibility and superior thermomechanical properties such as tensile modulus, failure strain, and toughness. This work offers two fully-developed, novel polymer platforms with outstanding performance; further, structure-property relationships are highlighted and discussed on a molecular and morphological level to provide material insights that are useful to researchers and engineers in the design of highly tuned and mechanically robust polymers.
Abstract
Although biochemical and physiological evidence suggests a strong interaction between striatal CB
1
cannabinoid (CB
1
R) and D
2
dopamine (D
2
R) receptors, the mechanisms are poorly ...understood. We targeted medium spiny neurons of the indirect pathway using shRNA to knockdown either CB
1
R or D
2
R. Chronic reduction in either receptor resulted in deficits in gene and protein expression for the alternative receptor and concomitantly increased expression of the cannabinoid receptor interacting protein 1a (CRIP1a), suggesting a novel role for CRIP1a in dopaminergic systems. Both CB
1
R and D
2
R knockdown reduced striatal dopaminergic‐stimulated
35
SGTPγS binding, and D
2
R knockdown reduced pallidal WIN55212‐2‐stimulated
35
SGTPγS binding. Decreased D
2
R and CB
1
R activity was associated with decreased striatal phosphoERK. A decrease in
mRNA
for opioid peptide precursors
pDYN
and
pENK
accompanied knockdown of CB
1
Rs or D
2
Rs, and over‐expression of CRIP1a. Down‐regulation in opioid peptide
mRNA
s was followed in time by increased DOR1 but not MOR1 expression, leading to increased D‐Pen2, D‐Pen5‐enkephalin‐stimulated
35
SGTPγS binding in the striatum. We conclude that mechanisms intrinsic to striatal medium spiny neurons or extrinsic via the indirect pathway adjust for changes in CB
1
R or D
2
R levels by modifying the expression and signaling capabilities of the alternative receptor as well as CRIP1a and the DELTA opioid system.
About Bach Butler, Gregory G; Stauffer, George; Greer, Mary Dalton ...
06/2008
eBook
That Johann Sebastian Bach is a pivotal figure in the history of Western music is hardly news, and the magnitude of his achievement is so immense that it can be difficult to grasp. In About Bach, ...fifteen scholars show that Bach's importance extends from choral to orchestral music, from sacred music to musical parodies, and also to his scribes and students, his predecessors and successors. Further, the contributors demonstrate a diversity of musicological approaches, ranging from close studies of Bach's choices of musical form and libretto to wider analyses of the historical and cultural backgrounds that impinged upon his creations and their lasting influence. This volume makes significant contributions to Bach biography, interpretation, pedagogy, and performance._x000B__x000B_Contributors are Gregory G. Butler, Jen-Yen Chen, Alexander J. Fisher, Mary Dalton Greer, Robert Hill, Ton Koopman, Daniel R. Melamed, Michael Ochs, Mark Risinger, William H. Scheide, Hans-Joachim Schulze, Douglass Seaton, George B. Stauffer, Andrew Talle, and Kathryn Welter._x000B_
CB1 cannabinoid receptors and the D2 dopamine receptors are both GPCRs that associate with interacting proteins (CRIP1a and DRIP) that bind to the C‐terminus of the receptor. The DRIP neuronal ...calcium sensor‐1 is involved in Ca2+ mediated regulation of the D2 receptor, whereas the recently discovered cannabinoid receptor interacting protein (CRIP1a), has been shown to suppress the tonic inhibition of voltage‐gated Ca2+ channels induced by CB1 (Niehaus et al., 2007). Therefore, CRIP1a may provide a new avenue for modulation of the endocannabinoid system.
AAV that contain transgenes to express shRNA designed to knock down CB1, D2R, or CRIP1a were created and injected via stereotaxically. Striata from AAV‐D2R and AAV‐CB1 shRNA‐treated rats displayed a significant reduction in D2 and CB1 mRNA levels, respectively. The AAV overexpressing CRIP1a virus increased CRIP1a mRNA levels without altering CB1 levels. However, significant reductions occurred in levels of preprodynorphin(Pdyn) and preproenkephalin‐1(Penk‐1).
In vitro, N18TG2 stably transfected with CRIP1a‐pcDNA3.1 displayed increased levels of CRIP1a, Pdyn, and delta opioid receptor mRNA levels without effecting CB1 levels. Additionally, CRIP1a overexpressers had a significant reduction in CB1 mediated phosphorylation of ERK during Rimonabant‐sensitive basal conditions. However, CB1 agonist mediated effects on maximum ERK phosphorylation were unaltered in CRIP1a overexpressors.
These studies provide both in vivo and in vitro insight into the mechanisms involved in CRIP1a modulation of CB1 receptors.
Attacks against computer networks are a serious threat and occur quite often. Currently there are methods using attack trees that can be used to model how these attacks may occur. We have extended ...this concept to a new tree structure called a protection tree as a tool for decision makers to allocate limited resources towards the appropriate defenses against a specified attack. Protection trees ensure these limited resources are used in a manner to achieve the highest probability of success in stopping an attack. Protection trees are produced systematically by first developing an attack tree, computing metrics for each node of an attack, and then developing a corresponding protection tree with similar metrics. Eventually, libraries of attacks and available protections can be used to automate the process of developing the trees. An example attack and protection tree is used to notionally show how an organization such as the department of homeland security can allocate resources to protect their computer networks from being compromised. Decision makers in the organization can use the resultant protection tree to determine where to allocate limited resources for the best protection of their network
In today’s highly competitive and challenging marketplace, manufacturing process improvement is more important than ever before. Conversely, it is probably also harder to achieve than at any time in ...the past. This is due to several factors. High levels of capital investment combined with short product life cycles mean that maximising utilisation levels of expensive equipment is essential. Increasingly complex production facilities are difficult to analyse and improve. The possibility of worsening the situation rather than improving it means that experimentation on the line itself is often a risk not worth taking. One solution to this problem is the use of computer based manufacturing system simulation. Simulation studies are beneficial because they remove the element of risk associated with experimentation. Potential process improvement strategies can be identified, evaluated, compared and chosen in a virtual environment before eventual implementation on the factory floor.<?xml:namespace prefix = o ns = "urn:schemas-microsoft-com:office:office" /> This research aimed to evaluate the use of discrete event system simulation in a real world manufacturing environment. To this end, a flexible simulation model of the main transfer line of Läpple Ireland, a large metal panel production facility, was designed and constructed using Extend simulation software. In conjunction with Läpple personnel, various ‘what if’ scenarios were identified and evaluated. These scenarios were aimed at deciding the best position for providing additional automation by investing in robots. From the results of the simulation modelling of the three main proposed modifications to the line, improvements of 9%, 18% and 33% in press line throughput were predicted. The negative effect on these improvements in the case that the proposed robots failed to achieve the desired speeds were evaluated. These negative effects were found to be not as dramatic as could be expected. The results were compared to those of similar research efforts elsewhere. Finally, future steps for the research to take were identified and suggestions for future areas of application for the model were made.
CB1 receptors are G-protein coupled receptors (GPCRs) abundant in neurons, in which they modulate neurotransmission. The CB(1) receptor influence on memory and learning is well recognized, and ...disease states associated with CB(1) receptors are observed in addiction disorders, motor dysfunction, schizophrenia, and in bipolar, depression, and anxiety disorders. Beyond the brain, CB(1) receptors also function in liver and adipose tissues, vascular as well as cardiac tissue, reproductive tissues and bone. Signal transduction by CB(1) receptors occurs through interaction with Gi/o proteins to inhibit adenylyl cyclase, activate mitogen-activated protein kinases (MAPK), inhibit voltage-gated Ca(2+) channels, activate K(+) currents (K(ir)), and influence Nitric Oxide (NO) signaling. CB(1) receptors are observed in internal organelles as well as plasma membrane. beta-Arrestins, adaptor protein AP-3, and G-protein receptor-associated sorting protein 1 (GASP1) modulate cellular trafficking. Cannabinoid Receptor Interacting Protein1a (CRIP1a) is an accessory protein whose function has not been delineated. Factor Associated with Neutral sphingomyelinase (FAN) regulates ceramide signaling. Such diversity in cellular signaling and modulation by interacting proteins suggests that agonists and allosteric modulators could be developed to specifically regulate unique, cell type-specific responses.
The present study investigated the effect of different levels of Delta-9-tetrahydrocannabinol (Delta(9)-THC) antinociceptive tolerance on Protein Kinase A (PKA) activity in mouse brain and spinal ...cord. To strengthen this investigation, a positive control was developed to demonstrate the assay utilized in this study was sensitive enough to detect an increase in PKA activity in the anatomical regions utilized in this study. The membrane-permeant and phosphodiesterase-resistant cAMP analog 8-Bromoadenosine-3',5'-cyclic monophosphorothioate, Sp-isomer (Sp-8-Br-cAMPS) was utilized for the development of this positive control and this compound produced an increase in PKA activity in several mouse brain regions (i.c.v.) and lumbar spinal cord (i.t.) following its administration. Models were then developed in which mice expressed either a 13-fold or 49-fold level of Delta(9)-THC antinociceptive tolerance following chronic treatment with 10mg/kg Delta(9)-THC or 80mg/kg Delta(9)-THC for 6.5 days. Basal and total cytosolic and particulate PKA activities were measured directly in homogenates from the striatum, hippocampus, cerebellum, cortex and lumbar spinal cord. Results from this study indicate that chronic exposure to Delta(9)-THC does not produce an increase in PKA activity in these mouse brain regions or spinal cord. Future work is needed to determine the role of PKA in cannabinoid tolerance in mice.
The present study investigated the effect of different levels of Δ-9-tetrahydrocannabinol (Δ
9-THC) antinociceptive tolerance on Protein Kinase A (PKA) activity in mouse brain and spinal cord. To ...strengthen this investigation, a positive control was developed to demonstrate the assay utilized in this study was sensitive enough to detect an increase in PKA activity in the anatomical regions utilized in this study. The membrane-permeant and phosphodiesterase-resistant cAMP analog 8-Bromoadenosine-3′,5′-cyclic monophosphorothioate, Sp-isomer (Sp-8-Br-cAMPS) was utilized for the development of this positive control and this compound produced an increase in PKA activity in several mouse brain regions (i.c.v.) and lumbar spinal cord (i.t.) following its administration. Models were then developed in which mice expressed either a 13-fold or 49-fold level of Δ
9-THC antinociceptive tolerance following chronic treatment with 10
mg/kg Δ
9-THC or 80
mg/kg Δ
9-THC for 6.5 days. Basal and total cytosolic and particulate PKA activities were measured directly in homogenates from the striatum, hippocampus, cerebellum, cortex and lumbar spinal cord. Results from this study indicate that chronic exposure to Δ
9-THC does not produce an increase in PKA activity in these mouse brain regions or spinal cord. Future work is needed to determine the role of PKA in cannabinoid tolerance in mice.
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